The alleged misconduct took place on a April 13 flight from Florida to California
This is Prudie Phones a Friend, a new feature where Jenée Desmond-Harris calls a few experts for their advice to a letter writer. Submit questions to Prudence here
“Brandon,” has been testing my patience lately
it’s become clear that he has a need for control and validation
Brandon recently confessed that he’s been deliberately leading on a man—flirting with him
and then pulling back whenever the man tries to take things further
admitted that he does it because he loves the ego boost it gives him
Nothing turns me on more than making someone want me who can never have me
Brandon wasn’t just confessing; he was bragging
He seemed proud of the way he was toying with this man’s emotions
and hearing Brandon talk about him in this way felt personal
It wasn’t just about Brandon’s toxic behavior; it was about the way he reduced this man to a pawn in his little power game
I wanted to call him out right then and there
I tried to explore why he feels the need to dominate others
but he just laughed it off and told me that it wasn’t that deep—he just found this guy fun to mess with
Every session with him leaves me drained and angry
and I’m starting to wonder if I’m the right person to help him
Maybe Brandon doesn’t really want to change
But then I wonder if perhaps I’m the one with the problem
It is not my job to only help clients I like
I have an ethical duty to help clients even if I wouldn’t associate with them in the real world
Maybe this is a learning experience for me that will make me a better therapist
Should I do the easy thing and drop Brandon as a client
Or should I take this as a challenge to put my personal feelings aside and help someone who clearly needs it
I think it’s a good sign that you’re paying attention to this
I imagine some less thoughtful people in your position might either say to Brandon
and is probably what Brandon needs to hear from a good friend
but wouldn’t solve the underlying problem) or tell him that due to a scheduling conflict
they wouldn’t be able to fit him in anymore
and that means you’re heading in the right direction
You need some instructions on how to move forward
You presented your options as A) working with Brandon despite taking his behavior and personality personally
and feeling drained and angry after your sessions
I felt a little stuck answering your letter because I did not think either of those choices would leave you feeling great
I can tell that you feel a sense of responsibility to be there for your clients
But it seems unsustainable —and not ultimately helpful to Brandon—for you to continue to meet with him while suppressing your negative feelings and working hard to keep your composure
I wasn’t sure if therapists had a framework for thinking about situations like the one you’re in and didn’t want to advise you in a way that would go against professional norms
So I reached out to Pamela LiVecchi, a clinical psychologist, professor, and the author of the book, Ethics for Counselors: Integrating Counseling and Psychology Standards
After noting that she appreciated your self-awareness and candor
“We’re all human and having reactions to one another is expected,” she told me that situations like yours
in which a therapist is bothered by their negative reactions to a client’s behavior or simply doesn’t like a client
there are a few actions to take before you decide to end the relationship
evaluate (and improve) the current state of your work situation
like when therapists have too many clients or lack support
it can be harder to feel empathetic,” LiVecchi said
I wonder if that kind of workplace sounds familiar
Is there anything you could do to slightly decrease your workload
or streamline your days so you feel less frazzled
The perspective you bring to your sessions with Brandon might differ depending on whether you’ve had a chance to eat lunch or are breathlessly rushing from one client to another
But you might have more capacity to hear about his antics and practice patience if you can decrease the stress in your workday overall
You should also consult with a trusted colleague about what you’re dealing with and how you’re feeling
“Therapists often consult when they’re struggling with their feelings about a client,” LiVecchi said
“We aren’t ethically obligated to help everyone
but we are obligated to behave ethically with everyone who we choose to work with
we’re determining if we have the necessary training and skills to be helpful
and if we’re going to be able to have a healthy professional relationship.” You don’t have to figure this out alone
Is there anyone in a similar role to yours whom you’d feel comfortable bouncing this issue off of (while still keeping patient confidentiality
Their perspective might help you clarify things
talking things through with a colleague isn’t enough
you might seek out professional supervision
“Feeling drained and angry after every session may be unhealthy for you and your client,” LiVecchi said
“Professional supervision can support deeper reflection on our emotional responses
it could help you better understand your reactions to your client’s behavior.” So consider turning to a more experienced therapist for guidance
Maybe you’ll even learn that they’ve been here before
you’ll need to evaluate whether therapy can be helpful to Brandon and whether you’re comfortable with your reactions to him
But if you conclude that Brandon isn’t open to much self-reflection or you are still having a strong negative reaction to him
another therapist with a fresh perspective may be best
you will be tasked with referring him to someone who can meet his needs—not just ghosting him
it won’t be an agonizing decision because you’ll know you’ve left no stone unturned when it comes to trying to make the relationship work
LiVecchi encouraged me to remind readers that if you’re wondering how your therapist feels about you
“This can lead to healthy and productive conversations that serve to strengthen the relationship and create a safe atmosphere,” she said
So maybe also have a quick answer ready just in case Brandon surprises you one day by asking
– The Irondequoit Town Police are looking for 62-year-old Gunther Powers
who they say is a vulnerable adult with dementia who might need medical attention
Police say Powers was last seen on foot along Brower Road in Irondequoit on Saturday at around 9:30 a.m
Police describe Powers as a bald man who weighs 200Ibs and is 5 feet 10 inches tall
They say he was last seen in a black hooded sweatshirt
call 911 or the Irondequoit Police Department at (585) 336-6000
Any person with disabilities who needs help accessing the content of the FCC Public file should contact Richard Reingold at rreingold@whec.com or 585-546-1701
There may not be obvious successors to the likes of Martin Amis and Salman Rushdie among today’s hotshot young writers
But is a new publisher dedicated to ‘overlooked’ male voices necessary
Who are this generation’s hotshot young male novelists
the modern incarnations of the Amis/McEwan/Rushdie crew of the 80s
The question flared again this week as writer Jude Cook launched a new press
Cook says the publishing landscape has changed “dramatically” over the last 15 years as a reaction to the “toxic male-dominated” scene of the 80s
excitement in publishing circles centres on a “new breed of young female authors
While this is “only right as a timely corrective”
the side-effect is that male authors are “often overlooked”
Some metrics certainly say so: of all the writers to appear on the weekly Sunday Times bestseller lists for fiction hardbacks so far this year
In nonfiction the picture is clearer: men make up 63% of authors to feature on the Sunday Times nonfiction hardback bestseller lists so far this year and 70% of those on the nonfiction paperback charts
with titles such as Atomic Habits by James Clear continuing to sell well years after publication
Writers and publishing insiders disagree over the extent of the gender imbalance in fiction
and the extent to which that imbalance is a concern
“There is no doubt male voices talking about male experiences – in middle age especially – are not getting much traction among agencies and publishers at the moment,” said one male agent, who asked to remain anonymous. However, men are “hardly underrepresented in the entertainment industry, and they need to earn their place as much as anyone else”.
Men “should make the work and let everyone decide if it’s worth anyone’s time, rather than pretend they are a minority in the arts”, says Sheena Patel, author of I’m a Fan. “I feel like men are doing just fine.”
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who runs the leading literary agency of the same name
“There was certainly a time when we needed more female novelists
particularly in the areas of crime and suspense
so I’m thrilled that female-led psychological suspense is still so popular
but the pendulum ultimately needs to land somewhere in the middle,” she said
While Milburn is “excited” about Conduit Books, she believes “we also need more male editors acquiring fiction in the areas men want to read”. The latest Publishers Association workforce survey suggests that 68% of publishing staff are women
Another female agent said that class may be a “bigger issue than gender when it comes to what’s being published”
Lacking racial representation also continues to be a problem: just one person of colour appeared on each of the Top 10 fiction and nonfiction hardback bestseller lists last week
Chimamanda Ngozi Adichie for fiction and Roxie Nafousi for nonfiction
said he knows “so many working-class men from my generation who got into reading fiction through Irvine Welsh
because it spoke to a world they recognised”
Welsh himself says that “men need to start reading before they start writing
My take is that men are becoming stupider because they over-rely on the internet and women are becoming smarter because they read more books.”
some suggest the decline of male novelists being commissioned is due to low demand from men
37% of fiction purchases in the UK were made by men
having made 58% of all book purchases last year
men buy more nonfiction (55% v 45% bought by women)
“Men need to read women and I think women should read men,” says Brown
“It’s one of the great ways we can come to understand each other better”
Cook’s new venture met with some criticism online
Mukherjee said that the project comes from a position that is “the opposite of Farage-ist grievance or misogyny-driven rabble-rousing.”
“No doubt there will be a lot of hand-wringing about this
“But writing and publishing are not zero-sum games.”
The state of Maine continues to defy President Donald Trump's executive order that bans males
whether they "identify" as girls and women or not
males continue to win girls' competitions.
On Friday, Soren Stark-Chessa won both the 800-meter and the 1,600-meter races at an event in Hiram, Maine, finishing the latter nearly 20 seconds ahead of the next-closest competitor
with Lilah Connor falling short of her male competitor by one-and-a-half seconds
Connor is seen shaking her head after coming in second place to a male athlete racing against girls.
Had Stark-Chessa competed in the boys' category
which would be the correct category since Stark-Chessa is male
the athlete would have finished in 12th place in the 1600-meter race and eighth in the 800-meter.
As OutKick's Amber Harding previously reported
this is not the first time Stark-Chessa has won girls' events
the male athlete is a Maine state champion.
"The runner won the 800-meter at the Maine Principals’ Association (MPA) Class C Girls’ Outdoor Track and Field State Championship as a sophomore in June
Stark-Chessa competed on the boys' teams," Harding wrote.
Last month, the United States federal government filed a civil lawsuit against the state of Maine for continuing to allow males to compete in women's sports
but that doesn't seem to have dissuaded the state from stopping it.
won two track and field events in Maine on Saturday
with the state currently being sued by the federal government
It appears that Maine, and its governor, are not interested in protecting fairness in girls' and women's sports on their own, and unfortunately, the only option left is to allow the legal system to do it for them.
Written by Dan ZaksheskeDan began his sports media career at ESPN
He is secure enough in his masculinity to admit he is a cat-enthusiast with three cats
one of which is named "Brady" because his wife wishes she were married to Tom instead of him
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(WHP) — A deceased male has been found by police after an extensive search detail in Lancaster County
according to the Lancaster County Coroner's Office
Officials state that a deceased male was found near a rail trail at the intersection of Landisville Road and Spooky Nook Road in East Hempfield Township
The coroner states that it appears that the man died by suicide
East Hempfield Police is the primary police agency on scene
The search detail was initiated around 4:00p.m
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as well as later life health conditions that occur in adults who were born after a complicated pregnancy
Researchers analyzed male and female placental samples from a larger study for differences in their methylation patterns
and found differences in gene activity between male and female placentas that may play a role in birthweight and adult diseases
The study identified 2,497 previously unreported DNA sites that had different methylation patterns for males and females
Of these and more than 2,500 sites that had also been identified by previous studies
66.9% of large increases in methylation occurred in DNA from male placentas and 33.1% from female placentas
Increases in methylation in male placentas was linked with greater neonatal size whereas those in females was linked with greater placental size
Some increases in methylation found in male placentas were located near the CCDC6 gene
Lower activation of CCDC6 has been linked with preterm birth in previous studies
inflamed skin) and higher risk for breast cancer later in life
The study was conducted by Fasil Tekola-Ayele
of the NIH’s Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD) and colleagues at other institutions
Tekola-Ayele F, et al. Sex-differentiated placental methylation and gene expression regulation has implications for neonatal traits and adult diseases Nature Communications. DOI: https://doi.org/10.1038/s41467-025-58128-3 (2025)
About the Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD): NICHD leads research and training to understand human development, improve reproductive health, enhance the lives of children and adolescents, and optimize abilities for all. For more information, visit https://www.nichd.nih.gov
National Institutes of Health (NIH)
NIH…Turning Discovery Into Health®
U.S. Department of Health and Human Services
The week of April 27-May 3 was filled with some outstanding individual performances from Des Moines metro-area high school male athletes
Boys track and field athletes had a productive week between the Drake Relays and state qualifying meets
1 West Des Moines Valley won a 1-0 battle over 3A No
Area boys golfers and tennis players also stepped up their game with the postseason just around the corner
Here are the Des Moines Register's male Athlete of the Week candidates from the Des Moines metro area for the week of April 27-May 3
DesMoinesRegister.com readers vote to determine the winners
who will be announced at the end of the week
ValleyMendez scored the winning goal for the 4A No
1 Valley boys soccer team during its 1-0 victory over 3A No
Mendez put the ball in the net off a feed from Eric Foust during the second half to give the Tigers the victory
Curtis led a balanced attack for the 4A No
2 Des Moines Roosevelt boys soccer team during its 10-0 victory over Waterloo East on April 29 at Waterloo
Curtis converted his only shot on goal into a score during the game
He also dished out one assists to help the Roughriders improve to 9-1-1 on the season
JohnstonKumaresan helped the Johnston boys tennis team prevail in both the No
1 singles and doubles matches during the Dragons' 9-2 victory at Dowling Catholic on April 29
Kumaresan defeated Dowling's Brady Frye in straight sets
1 doubles match he teamed with Aadil Patel to sweep past Frye and Rob Beecher with a pair of 6-3 victories
Haisman took medalist by four strokes for the North Polk boys golf team during the Winterset Invitational on May 2 at the Lakeview Country Club in Winterset
Haisman carded a 2-under-par round of 68 to defeat Perry's Daxon Weeks' score of 72 for the top individual score
Haisman also helped the Comets win the team title with a 302 to Winterset's 309
Des Moines EastParker helped the Des Moines East boys track and field team win the boys sprint medley relay at the Valley Relays on May 2 in West Des Moines
Parker anchored East to a winning time of one minute
The Scarlets finished comfortably ahead of Dowling Catholic's runner-up time of 1:34.80
Joe Randleman covers high school sports for the Ames Tribune. Contact him at jrandleman@gannett.com. Follow him on Twitter at @JoeRandleman
How would you feel if your coworker found your dating app profile and decided to treat you like you were desperate for trying to find love
that’s the situation this young woman is now in
She works in an industry that is mainly male
but she says that she adores her job and it’s her favorite one she’s ever held so far
One of her male coworkers recently came across her profile on a dating app
but he did mention his brother was the one who found it first and guessed what company she was at based on her location and industry
“I wasn’t working the day he made this discovery
he made it a point to let everyone know I was,” she explained
I then realized it was in a ‘…she’s desperate’ fashion
Not only is the workplace and field in general male-dominated
but I am more than 20 years younger than the majority of my coworkers
with the exception of the guy who spread the info and a couple of others.”
He and a few others bring it up in a mocking or teasing way quite regularly
It really wasn’t funny the first time
There’s no way her coworker is trying to be funny with how he’s going on about her dating app profile; he’s clearly doing this in a way intended to demean her
This is hardly the only nasty thing this man does
as he loves to give her advice she doesn’t ask for on her appearance
She’s wondering how on earth she can handle this quietly
as she does not want to go to HR and blow this up into a bigger mess than it already is
Sign up for Chip Chick’s newsletter and get stories like this delivered to your inbox.
Her plan is to change jobs in the middle of next year
so she wants to lay low until then and try to make the best of her time at her current company
“What should I say next time he or another coworker brings it up
More About:Relationships
ARIZONA NEWS
11:00 AM | Updated: 11:29 am
BY KTAR.COM
PHOENIX — A Phoenix high school teacher who was arrested in April for sex crimes against male students was indicted
the Maricopa County Attorney’s Office (MCAO) announced on Monday
Chapman’s arrest was prompted by a tip the Phoenix Police Department received from a social media provider via the National Center for Missing and Exploited Children
The tip said he had been enticing an account that had self-identified as a minor
Chapman, who is from Surprise, taught at Cortez High School, which is near 31st and Dunlap avenues, ABC15 reported
Court documents accused Chapman of filming himself sexually abusing a 16-year-old student in an alley near the boy’s home a week before his arrest
Chapman also allegedly sexually assaulted a 17-year-old victim on campus
Investigators found that Chapman also filmed himself having sex with a former student inside his current classroom
Chapman began sexually abusing him while he was a minor
Investigators also found Chapman saved various files depicting children being sexually abused
He allegedly claimed to prefer boys around 7 and 8 years old
Have a story idea or tip? Pass it along to the KTAR News team here
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Vote now for the Chillicothe Gazette high school male athlete of the weekChillicothe GazetteThis week's poll has athletes from baseball
A reminder to coaches/parents: If we do not have stats from games/matches
we can't put the athlete up for athlete of the week
Stats for any games/matches can be sent to cgosport@nncogannett.com after each event
please send complete stats to cgosport@nncogannett.com no later than 5 p.m
More: Glory Days: Chip Maxwell and the 1970 Zane Trace Pioneers
Last week's winner was Piketon's Wadye Fout
The week of April 27-May 3 was another eventful stretch of competition for Ames-area high school male athletes competing in spring sports
3 Ames boys soccer team defeated Des Moines Lincoln (4-0) and Des Moines Hoover (2-1)
scored a pair of 3-0 victories over Bondurant-Farrar and top rival Nevada
The Ames boys tennis team earned an 8-3 victory over Mason City on April 29 in Ames and the Ballard boys golf team defeated Adel-Desoto-Minburn
in a nine-hole dual at the Ballard Golf and Country Club in Huxley on April 29
golf and soccer teams also had a productive week
Area boys track and field teams continued to keep busy in the week between the Drake Relays and the state qualifying meets
Here are the Ames Tribune male Athlete of the Week nominees for the week of April 27-May 3. AmesTrib.com readers vote to determine the winner
3 Ames boys soccer team to a 4-0 victory over No
Sigurdsson put two goals on the board to help the Little Cyclones improve to 8-2-1 on the season
and he did an excellent job helping the Class 2A No
3 Gilbert boys soccer team dominate possession of the ball during a 3-0 victory over rival Nevada on May 1 at Gilbert
Mattson's goal came on a 25-yard direct kick with 21 seconds left in the first half
BallardColvin had a strong nine-hole round for the Ballard boys golf team during a 143-159 dual victory over Adel-Desoto-Minburn on April 29 at the Ballard Golf and Country Club in Huxley
Colvin was the individual medalist at the meet
He shot a 2-under-par round of 34 to win medalist by one stroke over teammates Mason Gatchel and Alex Weeks
Lucchesi gave the Roland-Story boys track and field team a win in the 3,200-meter run during the Norsemen Relays boys track and field meet on May 1 at Story City
Lucchesi won the 3,200 with a time of 10 minutes
Wierson won both the 100 and 200 dash events for the Collins-Maxwell boys track and field team at the South Hardin Invitational in Eldora on May 2
Wierson clocked in at 11.22 seconds in the 100 and 23.52 in the 200
MANSFIELD — It's time to make your vote count
Voting for the Mansfield News Journal Male Athlete of the Week is now open
CHANGES COMING: Everything is on the table: OHSAA looking at potential changes to football playoff format
Mansfield Senior's Will Fortner was voted the Mansfield News Journal Male Athlete of the Week last week with 3,406 votes
Fairfax, Va. – Diverse: Issues in Higher Education magazine named UMBC senior track and field student-athlete Jayo Adegboyo (Columbia
Diverse: Issues In Higher Education sponsors the Arthur Ashe Jr
Sports Scholar Awards to honor students of color who have excelled in the classroom and on the athletic field
Jr.'s commitment to education and love for the game of tennis
colleges and universities are invited to participate in this annual awards program by nominating their outstanding sports scholars
In addition to their athletic ability and academic performance
Sports Scholars demonstrate a commitment to community service and student leadership
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This week's winner will be recognized in the May 18 edition of The Advocate
Metrics details
The anterior cingulate cortex (ACC) is essential for social behavior
and its dysfunction is implicated in social interaction deficits in autism
Pyramidal neuron activity in the ACC is modulated by parvalbumin (PV) and somatostatin (SST) interneurons
though their specific roles in social interactions remain unclear
we demonstrate that PV and SST interneurons differentially contribute to the regulation of social interactions
is reduced in both PV and SST interneurons
Knocking out Kcnh7 in either interneuron subtype leads to social interaction deficits
projections from the lateral posterior thalamic nucleus (mediorostral part
LPMR) to PV interneurons and from the ventral hippocampus (vHPC) to SST interneurons differentially modulate social interactions
These findings provide new insights into the distinct roles of PV and SST interneurons in social processes and their contributions to autism-related pathophysiology
the precise mechanisms of ACC in regulating social interaction remain relatively understudied
complicating both the diagnosis and treatment of ASD patients
If PV and SST interneurons indeed selectively modulate ACC-related social processes
it is worth investigating whether they contribute to the heterogeneity of social deficits and exploring the underlying mechanisms
Using a Cre-dependent CRISPR-Cas9 viral approach to knockout Kcnh7 in PV or SST interneurons in the ACC
we found that PV interneurons primarily regulate sociability
whereas SST interneurons influence both sociability and social preference
The distinct contributions of PV and SST interneurons are partially due to their differential inputs
These findings provide direct evidence for the specific roles of ACC PV and SST interneurons in social interaction and offer insights into the potential mechanisms underlying social dysfunction in individuals with ASD
a Schematic diagram of AAV-FLEX-GCaMP7s injection (The 3D brain image was created with CEBSIT/ION Digital Brain https://www.digital-brain.cn/)
b Example image showing fiber placement in the ACC (left) and GCaMP7s expression (right)
d Cartoon depicting the social recognition behavior in the home cage test
e Ca2+ signals associated with social recognition in (d)
Right: the peri-event plot of the average Ca2+ transients: thick lines indicate the mean
shaded areas represent the SEM (n = 11 mice)
g Schematic diagram of the three-chamber sociability test
n = 11 mice) mice in social preference test
m Comparison of AUC (area under curve) (left) and the correlation index between social duration and neuronal activity in the PV-Cre and SST-Cre groups (right) (n = 11 mice per group
n Same as (m) but for sociability (n = 9 PV-Cre mice
o Same as (m) but for social preference (n = 9 PV-Cre mice
Source data are provided as a Source Data File
PV and SST interneuron activity was suppressed during social interaction
with PV interneuron activity changes being more prominent during sociability and SST interneuron changes being more pronounced during social preference
a Schematic diagram of virus injection (The 3D brain image was created with CEBSIT/ION Digital Brain https://www.digital-brain.cn/)
c Representative immunofluorescence images
d Number of PV+ cells (left) and Pvalb mRNA levels (right) (left: n = 6 slices from 2 mice per group
e Same as (d) but for SST-Cre mice (left: n = 6 slices from 2 mice per group
two-tailed unpaired separate variance estimation t test
P < 0.0001; right: n = 10 mice for the control and n = 15 mice for taCasp3
f Traces of PV-Cre mice before (left) and after (right) viral intervention in the sociability test
g Quantification of time spent in the social zone and empty zone before and after viral intervention for PV-Cre mice (n = 8 mice per group; two-tailed paired t test
P = 0.0097 for Pre; Two-sided Wilcoxon signed-rank test
h Discrimination score for PV-Cre mice in (g) (n = 8 mice
i Quantification of time spent in the stranger zone and original zone before and after viral intervention for PV-Cre mice (n = 8 mice
j Discrimination score for PV-Cre mice in (i) (n = 8 mice
k Same as (g) but for SST-Cre mice (n = 8 mice; two-tailed paired t test
m Traces of SST-Cre mice before (left) and after (right) viral intervention in the social preference test
n Same as (i) but for SST-Cre mice (n = 8 mice; two-tailed paired t test
o Same as (j) but for SST-Cre mice (n = 8 mice
a Schematic diagram of AAV- DIO-ChR2 injection into the ACC of PV-Cre mice and SST-Cre mice (left, the 3D brain image was created with CEBSIT/ION Digital Brain https://www.digital-brain.cn/) and diagram of the home cage test with laser delivery strategy (right panel is designed by us)
b Optogenetic stimulation of PV interneurons modulates social behaviors in the home cage test
Left: The raster plot of different types of social behavior in the home cage test; B body contact
Right: Statistical analysis of total social interaction time (n = 10 mice
c Same as (b) but for SST-Cre mice (n = 9 mice
d Comparison of slope coefficient in the PV-Cre
SST-Cre and Cre negative groups: X-axis: STON (social time when laser ON) -STOFF1 (social time during OFF1)/TT (total time of test); Y-axis: STOFF2 (social time during OFF2)- STON (social time when laser ON)/TT (total time of test) (n = 10 mice
e Comparison of the social inhibition index between the PV-Cre and SST-Cre groups (n = 10 PV-Cre mice
f Summary data of the time spent with toys for PV-Cre group in the OFF1
ON and OFF2 stages in the home cage (n = 10 mice
g Same as (f) but for SST-Cre group (n = 9 mice
The results indicate that both PV and SST interneurons modulate sociability in the home cage test
with PV interneuron activation exerting a stronger inhibitory effect on social behavior compared to SST interneuron activation
a Traces of PV-Cre mice in the sociability test
c Social interaction time (b) and discrimination score (c) from the sociability test (n = 9 mice
P = 0.008 for OFF1; two-tailed paired t test
P = 0.036 for ON; two-tailed paired t test
P = 0.161 for OFF2; two-tailed paired t test
d Traces of PV-Cre mice in the social preference test
c) but in the social preference test (n = 9 mice
P = 0.028 for ON; two-tailed paired t test
m Delay time when the first social behavior stopped during the sociability test (n = 9 PV-Cre mice
n = 8 SST-Cre mice; two-tailed unpaired t test
n Comparison of normalized discrimination scores in the sociability test (n = 9 PV-Cre mice
o Delay time when the first social behavior stopped in the stranger zone (n = 9 PV-Cre mice
q Time spent in the center zone for PV-Cre (p) and SST-Cre (q) in the social preference test (n = 9 mice
r Same as (n) but in the social preference test (n = 9 PV-Cre mice
n = 10 SST-Cre mice; two-tailed unpaired t test
These findings suggest that PV interneurons primarily regulate sociability
whereas SST interneurons have a more prominent influence on social preference in the three-chamber test
these results indicated that while both PV and SST interneurons contribute to sociability
SST interneurons play a more prominent role in regulating social preference
a RNAscope showing Kcnh7 (gray) and Pvalb (red) expression in WT (left) and Shank3 KO mice (middle)
with the quantification of Kcnh7+/Pvalb+ ratio (right
b Normalized fluorescence intensity of Kcnh7 (left) and normalized number of Kcnh7+ puncta (right) in Pvalb+ neurons (n = 13 cells in WT and n = 12 cells in KO from 3 mice per group; Mann‒Whitney U test
P = 0.003 for intensity; two-tailed unpaired t test
c Action potential trace at 200 pA in PV-tdTomato mice
d Input‒output curve (left) and spike frequency at 380 pA (right) (Stim
int.: stimulus intensity; n = 9 cells in WT and n = 10 cells in KO from 3 mice per group
Pwithin group < 0.0001 for input‒output; two-tailed unpaired t test
e Representative traces of the Kv11.3 current (left) and quantification (right) (n = 4 cells for the WT and n = 3 cells for the KO group from 2 mice per group; two-tailed unpaired t test
f Same as (a) but for Sst+ interneurons (right
g Same as (b) but for Sst+ interneurons (n = 11 cells for the WT and n = 12 cells for the KO from 3 mice per group
h Same as (c) but for SST-Cre::Ai9 neurons
i Same as (d) but for SST-Cre::Ai9 neurons (n = 9 cells in WT and n = 8 cells in KO from 3 mice per group
j Same as (e) but for SST-Cre::Ai9 neurons (n = 3 cells in WT and n = 4 cells in KO from 2 mice per group
a Time spent in the social zone and empty zone before and after viral intervention in PV-Cre mice during the sociability test (n = 9 mice
P = 0.008 for Pre; two-tailed paired t test
b Comparison of discrimination score for PV-Cre mice in the sociability test (n = 9 mice
c Time spent in the stranger zone and original zone before and after viral intervention in PV-Cre mice during the social preference test (n = 9 mice
P = 0.011 for Pre; two-tailed paired t test
d Comparison of discrimination score for PV-Cre mice in the social preference test (n = 9 mice
e Time spent in the social zone and empty zone before and after viral intervention in SST-Cre mice during the sociability test (n = 8 mice
f Comparison of discrimination score for SST-Cre mice in the sociability test (n = 8 mice
g Time spent in the stranger zone and original zone before and after viral intervention in SST-Cre mice during the social preference test (n = 8 mice
P = 0.012 for Pre; two-tailed paired t test
h Comparison of discrimination score for SST-Cre mice in the social preference test (n = 8 mice
a Schematic of experimental design (The 3D brain image was created with CEBSIT/ION Digital Brain https://www.digital-brain.cn/ and the right panel is designed by us)
b Representative images showing the starter neurons in ACC from a PV-Cre and a SST-Cre mice
c Representative images showing starter neurons in the ACC co-expressing AAV-helper (His-EGFP
and PV or SST (gray) (n = 6 slices from 3 mice per group)
d PV interneurons in the ACC receive direct inputs from the LPMR (top)
while SST interneurons in the ACC receive direct inputs from the vHPC (bottom)
Images are from representative animals of each group
e Quantification of principal starter neurons (expressing His-EGFP and RV-dsRed) in the ACC and PFC (n = 6 mice per group)
The value 1.00 indicates all detected starter neurons
f Analysis of RV-dsRed-positive cell density in upstream brain region (n = 5 mice per group)
Cl/DEn claustrum/dorsal endopiriform nucleus
HDB/VDB nucleus of the horizontal/vertical limb of the diagonal band
VL/AM ventrolateral thalamic nucleus/anteromedial thalamic nucleus
This study focused on two primary interneuron subtypes
We found that both types of interneurons significantly influence social behavior through distinct and nuanced mechanisms
we observed a reduced expression of the Kcnh7 gene
specifically in PV and SST interneurons within the ACC
Deletion of Kcnh7 in either PV or SST interneurons contributed to altered social interactions
highlighting the involvement of these interneuron subtypes in ASD-related behavioral variability
our pathway-specific analyses revealed that the LPMR-PV(ACC) and vHPC-SST(ACC) projections underline the roles of PV and SST interneurons in regulating social behavior
The LPMR-PV(ACC) pathway exerted a stronger influence on sociability
whereas the vHPC-SST(ACC) pathway predominantly affected social preference
our findings demonstrate that PV and SST interneurons in the ACC play a critical role in regulating social interactions through distinct synaptic inputs and downstream circuit mechanisms
the four strategies used to manipulate SST interneurons produced consistent effect on social preference
while their impact on sociability was more variable
These results suggest that SST neurons may be more actively involved in social preference
This conclusion is further supported by comparing the effects of PV and SST interneuron modulation
PV and SST interneurons showed reduced responses to both novel objects and conspecifics
but their suppression was stronger during interactions with novel mice
enabling differential regulation of pyramidal neuron responses to excitatory inputs
These distinctions allow PV and SST interneurons to fine-tune ACC activity during social information processing
contributing to the precise regulation of social behavior
While our study emphasizes specific projections to PV and SST interneurons
further research is needed to explore how upstream regions regulate other interneurons
Abnormal activation of PV or SST interneurons in the ACC could contribute to the aberrant social behavior
indicating that dysregulated upstream projections targeting these interneurons could be a critical factor contributing to such abnormalities
normal ACC activity during social recognition likely depends on the interaction between upstream projections and local circuits
Quantifying these contributions and developing mathematical models to describe the regulation of pyramidal neurons by PV and SST interneurons could elucidate their roles in social interaction and related behaviors
these findings highlight the complex interplay between molecular factors (such as Shank3 and Kcnh7)
cellular processes involving PV and SST interneurons
projection neurons and behavioral outcomes
This deeper understanding of how these elements interact offers valuable insights into the neurobiological basis of social dysfunction and its related disorders
the present research provides strong evidence that PV and SST interneurons in the ACC play differential roles in social interaction
These nuanced roles likely contribute to the various manifestations of social interaction deficits
This insight not only enhances our understanding of social behavior but also opens new avenues for addressing related disorders
By targeting the specific functions of PV and SST interneurons within the ACC complex network
novel therapeutic approaches may be developed underlying the new mechanisms of social dysfunction
017688) and C57BL6 mice were used in this study
The animals were housed under a 12 h:12 h light:dark cycle (lights on at 8 AM)
and no mice were excluded unless they exhibited weight loss exceeding 10% during subsequent experiments or if viral injections were performed inaccurately
as confirmed by post hoc analysis of viral targeting
The researchers were blinded to the experimental groups
All mice were euthanized using a humane method in accordance with the Principles of Medical Laboratory Animal Care
Six-to-eight-week-old mice were used for stereotaxic injections
The mice were anesthetized with 1% pentobarbital sodium and placed in a stereotaxic instrument (RWD)
Eye ointment and a piece of silver paper were applied to prevent corneal drying
A thermostatic heating pad (RWD) was used to keep the mouse body temperature constant during surgery
A small craniotomy hole was made using a dental drill (RWD)
and injections were carried out using a microsyringe pump (KD Scientific) at a flow rate of 40 nL/min to minimize brain tissue damage
Fiber photometry was used to record calcium signals via a commercialized fiber photometry system
240 nL/injection) was injected unilaterally into the ACC of PV-Cre or SST-Cre mice (AP: +0.73 mm; ML: −0.28 mm; DV: −1.75 mm)
optical fibers (2.5 mm optical density (O.D.)
2.5 mm length) were implanted ipsilaterally
We used dental acrylic resin (Super Bond) to support the ceramic ferrule
The mice were housed individually and allowed to recover for at least one week before subsequent experiments
AAV2/9-DIO-ChR2-EYFP was injected unilaterally into the ACC of PV-Cre or SST-Cre mice (virus titer: 3.2E + 12 vg/mL
240 nL/injection; AP: +0.73 mm; ML: −0.28 mm; DV: −1.75 mm)
2.0 mm length) were implanted ipsilaterally
A mixture of AAV2/9-CaMKII-GCaMP6s (5.36E + 12 vg/mL) and AAV2/9-hSyn-DIO-ChrimsonR-mCherry (2.50E + 12 vg/mL) at 1:8 ratio (200 nl total volume) was injected unilaterally into the ACC of PV-Cre or SST-Cre mice at the same coordinates as the previous site (AP: +0.73 mm; ML: −0.28 mm; DV: −1.75 mm)
The density of upstream neurons was analyzed with reference to the Mouse Brain in Stereotaxic Coordinates (Second Edition)
To trace synaptic connections from the LPMR to PV interneurons and from the vHPC to SST interneurons in the ACC
and to confirm the functional role of these projections
a mixture of AAVRetro-ChR2 (5.08E + 12 vg/mL) and AAV2/9-CAG-FLEX-GCaMP7s (2.21E + 12 vg/mL) was injected into the ACC of PV-Cre and SST-Cre mice (240 nL/injection)
Optical fibers (3.0 mm length or 4.0 mm length) were implanted in the ipsilateral LPMR (AP: −1.95 mm; ML: −1.64 mm; DV: −2.52 mm) or vHPC (AP: −3.2 mm; ML: −3.81 mm; DV: −1.57 mm) at a 15° angle
optical fiber (2.5 mm length) was implanted ipsilaterally in the ACC
To confirm the function of these two projections in vitro
AAV2/9-hSyn-ChR2-mCherry (3.84 E + 12 vg/mL
240 nL/injection) was injected into the LPMR or vHPC of PV-tdTomato or SST-Cre::Ai9 mice
The optical fibers were implanted in ACC ipsilaterally
For specific regulation of MD → PV interneurons
MD → SST interneurons and Cl/DEn→SST interneurons in the ACC
AAV1-Flp-EGFP was injected into the MD (AP: −1.0 mm; ML: −0.75 mm; DV: −3.4 mm)
Po (AP: −1.95 mm; ML: −1.25 mm; DV: −3.5 mm)
and Cl/Den (AP: +1.5 mm; ML: −0.85 mm; DV: −3.0 mm)
To suppress the activity of PV or SST interneurons and observe the impact on social behavior in mice
240 nL/injection) was bilaterally injected into the ACC
To specifically knockout Kcnh7 on PV and SST neurons
a mixture of AAV-MECP2-DIO-Cas9 (4.79E + 13 vg/mL) and AAV-kcnh7-gRNA-hSyn-GFP (1.55E + 13 vg/mL)
was injected bilaterally into the ACC of PV-Cre and SST-Cre mice
we did not define separate zero-points for each event
the zero-point was defined as the moment when the subject mouse approached the cage within a distance shorter than its head length
and signals were analyzed over a 5 s window following this point
The total observation time for both the home cage and three-chamber tests was 10 min
Changes in fluorescence intensity (ΔF/F0) were calculated as ΔF/F0 = (Fsignal–F0)/F0–Foffset
where F0 was defined as the baseline fluorescence signal averaged over a 2 s-long control time window and Foffset was the system bias data recorded in the dark
Fsignal represented the peak value of the fluorescence signal
Data analysis was performed using MATLAB-based software provided by Thinker Tech
For optical stimulation during behavioral tests, the device connection followed a previously described protocol7
which was linked to a fiber-optic rotary joint
The other end of the fiber was then connected to the ferrule mounted on the mouse’s head
Following a brief recovery period from handling
In the excitatory photogenetic experiments
the test was divided into three phases: laser stimulation off
Photostimulation of PV interneurons or SST interneurons in the ACC was induced by 40 Hz light trains with 5 ms pulses generated by a 473 nm laser (IOS-465
The laser power was adjusted to approximately 5.0 mW at the tip of the fiber-optic cable
Each phase (OFF1-ON-OFF2) lasted 3 min during both the home cage and three-chamber tests
divided by the total time of the test (TT); the Y-axis represented the social time after the laser was on (SToff2) minus the social time when the laser was on (STon)
divided by the total time of the test (TT)
Social inhibition index: the time spent with a social object when the laser was OFF1 minus when the laser was ON
divided by the total time spent with a social object when the laser was OFF1 and ON
To observe the effect of PV or SST neuron activation on the activity of pyramidal neurons in vivo
5 mW) was used to stimulate ChrimsonR-expressing interneurons
The stimulation protocol consisted of 10 cycles of 30 s ON followed by 1 min OFF during the home cage test
This experiment was performed as previously described7
The apparatus consisted of three identical chambers (30 cm × 45 cm × 20 cm) made of black perspex acrylic sheets
Two cylindrical grid cages were placed in the corners of the outer chambers
and auditory interaction between the test mouse and the caged mouse
the test animal was placed in the chamber for 10 min to habituate and then was isolated in the middle chamber to wait for the experiment to begin
a novel juvenile male mouse was placed in a wire cage in one chamber
and another empty wire cage was placed in the opposite chamber
The subject mouse was then allowed to explore all three chambers freely
the second stranger juvenile male mouse was put into the other empty cage and the experimental subject mouse was allowed to explore freely in three-chambers again
The data was analyzed via SMART v.3.0 software
The discrimination score in the sociability test was calculated as the time spent in the social zone minus the time spent in the empty zone
divided by the total time spent in both zones
The discrimination score in the social preference test was calculated as the time spent in the stranger zone minus the time spent in the original zone
The latency of the first social behavior stop was defined as the time from the start of the social interaction to the cessation of the behavior
When the subject mouse approached within a distance shorter than a mouse’s head length from the cage of the object mouse
time recording and optical stimulation were initiated manually
Laser activation was defined as the zero-point
and the end of the interaction was recorded when the subject mouse turned its body away from the cage or stepped back to initiate another behavior
Each trial included only one scenario in which the subject mouse entered a social zone
The experiment was conducted in an open field arena
Fresh urine was collected on the day of the experiment
Ten three-week-old male mice were placed in a cage lined with clean filter paper
the urine-stained portions of the filter paper were excised
Collected filter paper segments were placed into one dish
which was then covered with cling film punctured with a fine array of small holes
the filter paper was saturated with saline
The petri dishes with urine filter paper and saline filter paper were placed on opposite sides of the open field arena
The movement of the mice was recorded under two conditions: with the laser off and with the laser on
The time spent in each corner was analyzed using SMART v.3.0 software
a new round of social interaction experiments was conducted
The total observation time for each session was 10 min
The videos were analyzed via SMART v.3.0 software
Social promotion index: (social interaction time after viral intervention minus social interaction time before viral intervention) divided by (social interaction time after viral intervention plus social interaction time before viral intervention)
All animals were anesthetized with 1% pentobarbital sodium and perfused intracardially with 0.9% saline followed by 4% paraformaldehyde
The whole brain was then removed and placed in 30% sucrose solution for 3 days
40 μm-thick brain tissue sections containing the ACC were placed in blocking solution for 1 h before incubation with primary antibodies against PV (rabbit
brains were collected one hour after the behavior test
and tissue sections were processed as described above
These sections were stained with anti-Fos antibody (rabbit
The sections were incubated with the secondary antibody at a dilution of 1:800 for 4 h at room temperature
all the sections were incubated with Hoechst 33342 (Sigma) rinsed in 0.01 M PBS
Each section was mounted a microscope slide
and a cover glass was sealed on it with a mixture of 50% (v/v) glycerol in 0.01 M PBS and were then imaged with confocal (FV31S-SW
For the RV retrograde tracing of PV or SST interneuron monosynaptic inputs
7 days after RV-EnvA-ΔG-dsRed viral injection
the preparation of 40 μm-thick brain tissue sections was the same as described above
All the sections were incubated with Hoechst 33342 (Sigma)
Each section was mounted on a microscope slide and covered with a cover glass using a mixture of 50% (v/v) glycerol in 0.01 M PBS
The slides were then scanned using a VS200 Slide Scanner (Olympus)
The number of positive cells was analyzed via ImageJ software
and the area of the upstream brain region was measured via OlyVIA (Olympus)
The density of cells was calculated as the number of positive cells/the area of the upstream brain region
the mice were deeply anesthetized with isoflurane and sacrificed
Fresh ACC tissues were isolated from the corresponding coronal sections under a stereomicroscope
Total RNA was extracted with TRIzol reagent
and PrimeScriptTM RT Master Mix (Takara) was used to reverse transcribe the total RNA (500 ng) into complementary DNA
Real-time PCR was carried out via a StepOnePlusTM Real-Time PCR instrument (Thermo) with 2× Universal SYBR Green Fast qPCR Mix (ABclonal)
The sequences of the primers used were as follows:
mice were deeply anesthetized with isoflurane and transcardially perfused with ice-cold carbogenated (95% O2
5% CO2) cutting solution containing 115 mM choline chloride
Coronal sections (300 µm thick) containing the ACC were cut using a vibratome (7000SMZ
the brain slices were incubated with carbogenated cutting solution (95% O2
the slices were incubated for 1 h at room temperature in artificial cerebral spinal fluid (ACSF; 119 mM NaCl
the slices were placed in the recording chamber and continuously perfused with ACSF
Recordings were obtained using an Axon Multiclamp 700B amplifier (Molecular Devices) filtered at 5 kHz and sampled at 20 kHz with a Digidata 1550B
Clampex 10.6 and Clampfit 10.7 were used for acquisition and analysis
Data were excluded if the resting membrane potential of neurons were more positive than −60 mV
or the series resistance changed by >20%
To record light-induced spikes with a current clamp in PV-positive and SST-positive neurons in the ACC
patch pipettes were filled with a solution containing the following: 128 mM potassium gluconate
In the current clamp mode of the whole-cell patch
5 ms pulse width and 40 Hz 473 nm light stimulus (IOS-465
RWD) was delivered to evoke spikes during each stimulus
The Event Detection function in the Clampfit software was used to quantify the number and amplitude of the spikes
Patch pipettes were filled with a solution containing 103 mM CsCl
The membrane potential of pyramidal neurons was held at −65 mV during laser-evoked IPSCs and PPR recordings
PV or SST interneurons were stimulated with a 5 ms pulse of 473 nm light (IOS-465
100 μM) was added to verify the properties of the IPSCs
To evaluate the intrinsic membrane properties of PV interneurons and SST interneurons in the ACC
whole-cell recordings were performed in current-clamp mode
and spikes were elicited by incrementally increasing the current injection in 20 pA steps
Patch pipettes were filled with an internal solution containing the following 120 mM CsMeSO3
The laser intensity was adjusted to achieve a maximal response without overstimulation
and 0.5 ms pulses of blue light was applied with the cells clamped at −65 mV
An additional 1 μM tetrodotoxin (TTX) and 100 μM 4-AP were added
to determine whether the light-evoked PSCs were excitatory or inhibitory
To investigate the role of the Kv11.3 channel in action potentials
1-[2-(6-methyl-2-pyridyl)ethyl]-4-(4-methylsulfonylaminobenzoyl) piperidine (E-4031
Stock solutions of E-4031 (100 µM) were prepared in DMSO and diluted in ACSF
ensuring that the final DMSO concentration did not exceed 0.1%
which had no detectable effect on the membrane potential or membrane currents
To record Kv11 currents from PV or SST-positive neurons in the ACC
50 μM D-AP5 and 10 μM bicuculline were added to high-K+ intracellular solutions (10 mM KCl and no Ca2+)
The use of high-K+ extracellular and low-K+ intracellular solutions allowed the K+ currents to be recorded as large
The signals were filtered at 3 kHz and digitized at 10 kHz in experiments for Kv11 current measurements
The activation curve of the Kv11 channels was extracted by dividing the peak amplitudes of the E-4031-sensitive tail current traces at 0.5 ms after the voltage was stepped to −100 mV
ACC tissues were rapidly dissected from the cortices of Shank3 KO and WT mice using freshly frozen mouse brain samples
and sequencing were performed by Gene Denovo Biotechnology Co
China) following the 10× Genomics (USA) guidelines
the tissues were homogenized in ice-cold homogenization buffer (0.25 M sucrose
The homogenates were sequentially filtered through a 70 µm cell strainer to isolate the nuclear fraction
The nuclear fraction was mixed with an equal volume of 50% iodixanol and carefully layered onto a discontinuous gradient of 30–33% iodixanol
This mixture was centrifuged at 10,000 × g for 20 min at 4 °C
After removing the myelin layer from the top of the gradient
nuclei were harvested from the interface between the 30% and 33% iodixanol layers
The isolated nuclei were resuspended in nuclei wash and resuspension buffer (0.04% bovine serum albumin
and 0.1 mM PMSF protease inhibitor in phosphate-buffered saline) and pelleted by centrifugation at 500 × g for 5 min at 4 °C
To remove cell debris and large aggregates
the nuclei suspension was further filtered through a 40 µm cell strainer
The nuclei concentration was quantified using trypan blue exclusion staining and a hemocytometer and adjusted to 800–1200 nuclei/µL
and library preparation were performed according to the 10× Genomics protocol
Cellular suspensions were loaded onto a 10× Genomics GemCode single-cell instrument
which generated single-cell Gel Bead-in-EMulsion (GEMs)
Libraries were generated and sequenced from the cDNA using Chromium Next GEM Single Cell 3′ Reagent Kits v3.1
The cell-by-gene matrices were produced via UMI counting and cell barcodes calling
The resulting matrices were individually imported into Seurat (version 3.1.1) for downstream analysis
The expression levels of each gene in a given cluster were compared against those in the remaining cells using the Wilcoxon rank sum test60
The mice were deeply anesthetized with isoflurane and transcardially perfused with 0.01 M PBS (pH: 7.4)
followed by 4% paraformaldehyde in 0.01 M PB
Brains were removed and fixed in 4% paraformaldehyde for 2 h and then sequentially incubated in 10%
The tissues were frozen and cut into 10 μm thick coronal sections using a cryostat (Leica)
RNAscope in situ hybridization was performed using the RNAscope™ Multiplex Fluorescent Reagent Kit v2 (ACD) for fixed-frozen tissue following the manufacturer’s instructions
the sections were dehydrated in sequential incubations with different concentrations of ethanol
followed by 30 min of Protease III treatment and washing in ddH2O
with each target incubated with a specific dye
Hoechst counterstaining and mounting were performed with antifade mounting medium
The fluorescence intensity was analyzed via ImageJ software
The total number of puncta was used to normalize mRNA expression
Single PV-positive or SST-positive neurons in the ACC with red fluorescence signals were aspirated using a patch clamp
Each single cell was ejected into a tube with 5 µL of 0.01 M D-PBS
Total RNA and cDNA from single neurons were isolated using a REPLI-g® WTA single-cell kit (Qiagen) according to the manufacturer’s instructions
qRT‒PCR was performed with SYBR® Premix Ex Taq (ABclonal) in a CFX96 Touch Real-time PCR Detection System (Bio-Rad)
The mean expression value of the control group was calculated
The data of the experimental group and control group were divided by this mean
and the resulting values were used for statistical analysis
An AAV CRISPR/Cas9 approach was used to conduct the in vivo Kcnh7 knockdown experiments
Single guide RNAs (sgRNAs) with high specificity and high efficiency were computationally identified in sgRNA libraries for genome-wide CRISPR knockout screening
Three U6-sgRNA (FE) gene fragments with the F + E tracrRNA backbone were synthesized by Integrated DNA Technologies (sequences are provided below
These fragments were subsequently cloned and inserted into the pX552-EGFP plasmid via In-Fusion Snap assembly master mix (Takara
resulting in the construct pX552-3xsgRNA(FE)-EGFP
The AAV vectors were serotyped with PHP.eB coat
plasmid #112867) and pUCmini-iCAP-PHP.eB (Addgene
plasmid #103005) were cotransfected into HEK293T cells using polyethylenimine (Cat
the cells were harvested by centrifugation at 5000 × g for 10 min at 4 °C
The AAV vectors in the media were precipitated with 8% PEG8000 (89510-1KG-F
The cell pellets and vectors precipitated from the media were resuspended in digestion buffer containing 500 mM NaCl
Novoprotein) was added to the digestion buffer
and the mixture was incubated at 37 °C for 1 h
the mixture was centrifuged at 2000 × g for 10 min
and the supernatant was added to a 39 mL ultracentrifuge tube with discontinuous gradients of 15%
Ultracentrifugation was performed at 350,000 × g for 2.5 h at 18 °C
Fractions (5 ml) from the 40% layer and 40–60% interface were collected and desalted using a 100 kDa cutoff ultrafiltration tube (15 ml
The buffer was exchanged four times with 1× PBS containing 0.001% Pluronic F-68
AAV titers were determined via qRT‒PCR using the primers for ITR
Forward primer: 5′-GGAACCCCTAGTGATGGAGTT-3′; Reverse primer: 5′-CGGCCTCAGTGAGCGA -3′
gagggcctatttcccatgattccttcatatttgcatatacgatacaaggctgttagagagataattggaattaatttgactgtaaacacaaagatattagtacaaaatacgtgacgtagaaagtaataatttcttgggtagtttgcagttttaaaattatgttttaaaatggactatcatatgcttaccgtaacttgaaagtatttcgatttcttggctttatatatcttgtggaaaggacgaaacaccgCTGGGCAATATCATGCCTCTgtttaagagctatgctggaaacagcatagcaagtttaaataaggctagtccgttatcaacttgaaaaagtggcaccgagtcggtgct
AaggtcgggcaggaagagggcctatttcccatgattccttcatatttgcatatacgatacaaggctgttagagagataattagaattaatttgactgtaaacacaaagatattagtacaaaatacgtgacgtagaaagtaataatttcttgggtagtttgcagttttaaaattatgttttaaaatggactatcatatgcttaccgtaacttgaaagtatttcgatttcttggctttatatatcttgtggaaaggacgaaacaccGAATATCGGGTTGACCCTCTCgtttaagagctatgctggaaacagcatagcaagtttaaataaggctagtccgttatcaacttgaaaaagtggcaccgagtcggtgct
AaggtcgggcaggaagagggcctatttcccatgattccttcatatttgcatatacgatacaaggctgttagagagataattagaattaatttgactgtaaacacaaagatattagtacaaaatacgtgacgtagaaagtaataatttcttgggtagtttgcagttttaaaattatgttttaaaatggactatcatatgcttaccgtaacttgaaagtatttcgatttcttggctttatatatcttgtggaaaggacgaaacaccgATGCCCTCCGTATGCTACAGgttttagagctagaaatagcaagttaaaataaggctagtccgttatcaacttgaaaaagtggcaccgagtcggtgc
Statistical significance was considered when p < 0.05
Further information on research design is available in the Nature Portfolio Reporting Summary linked to this article
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Guoping Feng (Massachusetts Institute of Technology) for his suggestions and conversation
This study was supported by the Natural Science Foundation of China (32394032 to S.W.)
the Science and Technology Innovation 2030 Project of China (2021ZD0201005 to S.W.)
the Natural Science Foundation of China (82221001 to S.W.
the Shaanxi Provincial Key Research and Development Program (2020ZDLSF01–09 to S.W.)
the Shaanxi Provincial Innovation Chain Project of Key Industries (2023-ZDLSF-47 to W.W.)
the Shaanxi Province Key Research and Development Plan (2024SF-YBXM-033
the Joint Founding Project of Innovation Research Institute
the Shanghai Pujiang Program (22PJ1415500 to Q.C.)
the Zhongshan Municipal Bureau of Science and Technology Grant (CXTD2022013 to Q.C.)
Guangdong Province High-level Innovative Research Institute project (2021B0909050003
and Innovation Team Accredited by Shaanxi Science and Technology (2022TD-49 to W.W.)
We thank Analysis & Testing Laboratory for Life Sciences and Medicine of Fourth Military Medical University for their assistance with behavioral test
We thank American Journal Experts for language editing (Certificate NO: 9183-20FC-2A00-204D-DD6P)
These authors contributed equally: Chuchu Qi
performed the experiments and analyzed the data
The authors declare no competing interests
Nature Communications thanks the anonymous reviewers for their contribution to the peer review of this work
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England’s governing body for professional and amateur soccer will no longer allow transgender-identifying males to compete in women’s soccer starting next month
The English Football Association (EFA) said the change will take effect on June 1
It comes in response to a United Kingdom Supreme Court ruling last month that the definition of woman under the Equality Act of 2010 is based on anatomical sex
“As the governing body of the national sport, our role is to make football accessible to as many people as possible, operating within the law and international football policy defined by UEFA and FIFA,” EFA said in a statement.
which allows transgender women to participate in the women’s game
was based on this principle and supported by expert legal advice.”
EFA expressed empathy for transgender-identifying people and said it hopes to keep them involved with the sport.
and our position has always been that if there was a material change in law
or the operation of the policy in grassroots football then we would review it and change it if necessary,” EFA said.
“We understand that this will be difficult for people who simply want to play the game they love in the gender by which they identify
and we are contacting the registered transgender women currently playing to explain the changes and how they can continue to stay involved in the game.”
EFA isn’t the only sporting body in the United Kingdom to change its transgender athlete policy since last month’s ruling. The Scottish Football Association did the same last week
and the U.K.’s Ultimate Pool changed its policy the week prior.
“We respect that some people within the pool community may find the changes challenging,” Ultimate Pool posted on X last month
we are committed to being empathetic to all members of our community and we expect all members of our community to reflect this.”
Ultimate Pool made the change after Harriet Haynes and Lucy Smith
competed against each other in the championship round of its Women’s Pro Series event last month
Haynes defeated Smith and got the British equivalent of $2,300 in prize money.
Protesters at the event held signs saying, “Save women’s sport” and “He’s a man,” as The Lion reported.
A February 2025 YouGov poll found 74% of Britons oppose letting transgender-identifying male athletes compete in women’s sports
A school sports coach is seeking advice after being invited to a pool party by his team
While he believes that the get-together is inappropriate and uncomfortable
the teachers and parents do not see the issue
he is wondering how he should politely decline the invitation without making it into a big deal
or is he right to feel uncomfortable about attending
Sharing his predicament to the subreddit r/Teachers
the coach admitted that he found it bizarre that none of the parents or teachers believed that extending the invite to him would be inappropriate
he feared that sharing his concerns would land him in further trouble
“My main problem is I feel even saying that it may be inappropriate for me to come to the party would make the parents think
'Why is it inappropriate?’ Which then makes me look like I have a perverted mind.” He explained that last year’s get-together was not an issue
given that the team invited him bowling and no one was in a bathing suit
RELATED: Male Teacher Is Uncomfortable With School’s New Rule To Close & Lock The Classroom Door At All Times — Even If There's One Female Student In The Classroom With Him
but they should know in today's age how this would be inappropriate.” The coach also feared that if he attended the pool party
word would spread among other school parents who might complain
He asked others what the best approach to the situation would be
“I am a male and I coached 7th and 8th grade girls' volleyball for 7 years. I would thank them for the offer and kindly decline
You don't need to give a reason,” one Redditor suggested
We understand and will follow suit,” another wrote
“I’m a male coach of a high school girls team
I got invited to a grad party last year that was a pool party
and got there right when it started when no one was in the pool let alone in bathing suits yet
Had a great time and left,” another user shared
While no one would argue that the coach's presence is no different than if he were at a public pool or at a beach, but it's important to remember that even though he is not technically working during the party, he is still there in a professional capacity. That means he needs to put his coach hat on and keep his workplace boundaries in place
Kevin Chapman, Ph.D., clinical psychologist and founder of The Kentucky Center for Anxiety and Related Disorders, told True Sport
we have unrealistic expectations for coaches
people expect coaches to parent their kids
Coaches are responsible for enhancing the development of the student athlete by teaching them discipline
which are those skills that they won’t learn necessarily in other settings
And that’s why being an athlete is so incredibly rewarding: If you have the right coaches
None of those expectations for coaches include attending a pool party that makes their relationship seem more familiar than it should
RELATED: Mom Expresses Concern After Her 7-Year-Old Daughter Drove To School Alone In A Car With A Male Teacher She’s Never Met
While extending the invitation to the coach was a kind gesture
a pool party is not exactly the right setting
He needs to do what he feels is right for him as a professional coach and as a man
If his team wants to thank him for a great season, it would be better to organize a casual team dinner where his boundaries and comfort would not be in jeopardy
it is a good thing that the coach recognized the bizarre nature of the setting for a team gathering
Not only does he have integrity that makes him a good person
but it most likely makes him one great coach that his team — and their parents — can trust
RELATED: Male Substitute Teacher Is Fired After A Student Hugs Him
Megan Quinn is a staff writer with a bachelor's degree in English and a minor in Creative Writing
She covers news and lifestyle topics that focus on justice in the workplace
© 2025 by Tango Publishing Corporation All Rights Reserved
Class of 2026 catcher prospect Francisco Rivero was recently selected the 5A Sonoran Player of the Year after powering Canyon del Oro to an impressive 10-game winning streak to end April
Rivero went an incredible 18 for 29 (.621) with five home runs and 23 RBIs in that 10-game stretch that was key to the Dorados’ current 24-5 record and being in the second round of the 5A state playoffs
in a 10-6 win over Catalina Foothills on April 22
Rivero is the Jet Sports Training Male Athlete of the Month
The former Sunnyside Little League standout
who was part of the team that came a win away from the Little League World Series in 2019
spent his first two years of high school in the Dominican Republic training with the hope of being selected in the international draft
The family chose to have him finish his high school experience in Tucson and potentially either play in college or go to a major-league organization after graduating from CDO next year
Rivero is batting .513 overall with seven home runs and 36 RBIs
FOLLOW @JAVIERJMORALES ON TWITTER
He became an educator five years ago and is presently a special education teacher at Gallego Fine Arts Intermediate in the Sunnyside Unified School District
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Georgia Gov. Brian Kemp on Monday signed into law a bill that prohibits males who claim to be females from participating in women’s and girl’s sports. The measure passed the Republican-controlled state House and Senate largely along party lines with a total of just five Democrats supporting the bill
The new law bars males from participating on any K-12 or collegiate level female-only sports teams
Girls are also prohibited from participating on male teams unless a corresponding female team does not exist
private schools that participate in competitions
and higher education institutions are required to separate multiple occupancy restrooms and facilities by sex
Dig deeper: Read Christina Grube’s report about the U.S
House of Representatives sending a subpoena to fencing officials in regards to athletes who call themselves transgender
If you enjoyed this article and would like to support WORLD's brand of Biblically sound journalism, click here
Lauren Canterberry is a reporter for WORLD
She graduated from the World Journalism Institute and the University of Georgia with a degree in journalism
She worked as a local reporter in Texas and now lives in Georgia with her husband
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Boyd McWhorter Male Scholar-Athlete of the Year4/30/2025 | Baseball
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I wouldn’t be the best at beard maintenance
I cut a few hairs that refuse to do anything but stand horizontally or vertically out from my face
Sometimes I look like I’ve been receiving mild electric shocks while I was asleep
In the bathroom I have a little beard brush yoke and a dust buster which I bought to get rid of the detritus and to keep my marriage stable: hairs are a contentious issue
the beard will take on an odd shape and I’ll start to look like that cliched image of a crazy old fellah who has been trapped alone on a desert island for years
It always strikes me as somewhat miraculous that a barber
can transform the unruly mess on my chin into something neat and pretty presentable; and do it while having shouted conversations with some of the other people there
I tend not to engage in this talk because, obviously, I have to keep my chin still, but also because – in my experience – the conversations are almost exclusively about football
about which I have an almost total ignorance
an entry method for serious discussions or gentle joshing
it also seems to be a way of comforting the clients
Because the clients are vulnerable: sitting powerless while another man deploys razor-sharp implements around them
The clients also have to reveal thoughts about their own appearance: how they want their hair or beard
talking about this kind of thing might make some men uncomfortable
They might not seem to be obviously connected
but barber shops make me think of record shops
I don’t mean the big high-street multiples flogging T-shirts
games and over-priced vinyl reissues of Dark Side of the Moon and Rumours
where at least half the stock is second-hand
and it’s taken on trust that these records will be in playable condition
These kinds of places are all over country
and it’s become a habit for me to visit the good ones in various towns and cities
the owners of these shops may well object to that depiction: they’ll argue that their clientele consists of all ages and genders
But I would guess that their core cohort of customers is made up of men of a certain age
The proprietors of these shops – in my experience – are all men like me
But that’s not a bad thing. I go in and have a bit of a dig around. Sometimes, the owner is having a conversation with a customer. It’s usually, though not exclusively, about music
the owner may well have a short chat with me as well
[ ‘That’s a Tesla,’ my daughter said. ‘We don’t like them’Opens in new window ]
And I always find the experience comforting. The phrase “male space” can come with all sorts of negative connotations, and for all sorts of justifiable reasons. Yet that’s not always the case.
Young men are starved of role models. And the few who wish to promote themselves as exemplars all too often want to define “maleness” as something that is purely oppositional: being a man means being angry at the shape of the world. But there are alternatives. Go for a haircut, buy a few records. You could do a lot worse.
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IFLScience HomeSperm-Blocking Male Contraceptive Could Hit The Market "As Soon As 2028" After Promising 2-Year Trial ResultsComplete the form below to listen to the audio version of this article
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the developers say it blocks sperm transit without affecting ejaculation or sensation
The makers of a non-hormonal male contraceptive just announced a key milestone during their first human clinical trials
with the treatment demonstrating safety and efficacy at the two-year mark
believe their product could come to market before the end of the decade
The treatment is called ADAM™
and it works by temporarily blocking the transit of sperm through the vas deferens
the long muscular tube that connects the testes and urethra.
The injectable hydrogel, administered within a 30-minute appointment
prevents sperm movement without blocking ejaculation or affecting sexual sensation
the gel will harmlessly liquefy and sperm flow will be restored
as Contraline CEO Kevin Eisenfrats told IFLScience
“No other method can provide long-lasting (two years)
I believe we're now closer than ever to figuring out the science piece
for those in possession of a male reproductive system
There are even other approaches similar to ADAM™
These approaches are appealing for the same reasons as the female contraceptive implant or IUD – once it’s in place
you theoretically don’t have to think about it again for months or even years
There’s no remembering to take a pill every day
Eisenfrats believes that even within this field
“Other vas-occlusive methods are designed to be permanent and require a second procedure to remove the implant
the harder it becomes to successfully reverse
Vas-occlusion is nice in that it is 'set and forget' and the mechanism of action is to block sperm right at the source without affecting anything else in the body,” he told IFLScience
Ultimately, the goal would be for male and female contraceptives to be available in a similar range of choices
“We understand that not every guy may want to get a procedure
“We need to offer men a bunch of options.”
Contraline’s latest announcement is an update on the first-in-human trials of ADAM™ now that some patients have reached the two-year milestone
Two patients have been shown to have azoospermia – an absence of viable sperm in their semen – 24 months after the administration of the hydrogel
Other patients who were recruited later are still in the trial
with successful contraceptive efficacy being shown at 12
No participants have experienced serious adverse effects from the treatment
meaning the study team is confident in progressing to Phase 2 trials
Eisenfrats told us they hope to recruit 30 people at a minimum
“We believe ADAM™ will be available by the end of this decade
and potentially as soon as 2028,” he said.
Recruitment is unlikely to be a major barrier
if the interest in Contraline’s products is any indication
“[We] have a waiting list of 15,000+ men (without any advertising/marketing) and what we have found is that a large percentage of men want a quick and minimally invasive procedure like ADAM™ that offers years of non-hormonal birth control,” Eisenfrats told IFLScience
If ADAM™ were to progress successfully through all the stages of a clinical trial
“Researchers have been working on male contraception for 60+ years
but there has never been a method that has shown full safety
and reversibility beyond Phase 2 trials,” said Eisenfrats
“With recent advancements in drug development and medicinal chemistry
I believe we're now closer than ever to figuring out the science piece.”
maybe now is the right time – perhaps ADAM™ will be the treatment that finally breaks the deadlock and sets us on the path to a greater range of male contraceptive options
“[I] also do believe men are more ready for new male contraceptives now than they were 20
There's no question that there is demand from consumers.”
medicineRFK Jr Calls For Placebo Trials For All New Vaccines. Experts Say You Can’t – And Shouldn’t – Do That1 hour agolink to article
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but revolutionary test has been developed to help unravel the mystery of infertility among men who display normal semen parameters
Infertility affects an estimated one in every six couples worldwide with male factor prevalent in about half of those cases
For many of those men who have seemingly otherwise healthy sperm the anguish of their condition is deepened by the perplexity of an underlying reproductive dysfunction called CatSper-related male infertility
there are defects in the chemosensory CatSper channels of sperm that allows them to move in a whip-like fashion essential to penetrate the coating of a female egg
There are many case histories in fertility treatment of couples where conventional IVF and intrauterine insemination (IUI) has failed
but where single sperm injection of an egg – known as intracytoplasmic sperm injection
The 2025 Congress of the Asia Pacific Initiative on Reproduction (ASPIRE) in Singapore today heard of a breakthrough diagnostic test for CatSper-related male infertility that has delivered remarkable validation results
a pathfinding researcher in the Centre of Reproductive Medicine and Andrology at the University Hospital Münster in Germany
said a desire to better understand the role of CatSper in male fertility led to the development of the test as a novel diagnostic tool
"Sperm generate a powerful swimming behaviour known as hyperactivation
a specific calcium channel found exclusively in the tail of sperm
sperm cannot hyperactivate and the egg remains unfertilised
"The CatSper test is a simple procedure with a standard semen analysis
It quickly and reliably determines whether a CatSper defect is present
enabling evidence-based therapy choices for couples with unexplained infertility
"We used it to screen 2,386 patients as a proof-of-concept study
It showed exceptional sensitivity and specificity of the test with precise assessment of CatSper function
"It has transformed the CatSper test from a scientific possibility into a diagnostic device for everyday clinical practice
"For the first time it allows diagnosis of CatSper defect in male infertility potentially preventing couples from embarking on unsuccessful medically assisted reproduction attempts such as IUI and IVF
"This has the potential to save patient expense and possible adverse effects of hormonal stimulation in IVF for the female partners
"The time factor of going through cycles of IVF is also an important consideration for couples striving to conceive
especially when almost every day counts among women in their thirties and forties when fertility is in decline
"The CatSper test can guide clinicians toward more personalised
The ASPIRE Congress in Singapore has attracted around 2,000 scientists
nurses and counsellors in fertility health to address latest global advances in assisted reproduction
★ Poor ★ ★ Promising ★★★ Good ★★★★ Very good ★★★★★ Exceptional
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UVA Alert Monday morning from self-inflicted gunshot incident
The Nelson County Sheriff and The AWARE Foundation of Virginia asking help finding a missing Nelson County teen.
On Friday, Governor Youngkin took final actions to complete the legislative session for 2025 in the General Assembly.
Charlottesville Police Chief Kochis presented to City Council last night a plea to implement a one-year program that would combine department program data, asking Council for a $150,000 appropriation.
Virginia’s standing as one of the top states in the nation for patient safety in hospital settings has again been affirmed in the Leapfrog Group Hospital Safety Grade rankings.
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The Hidden Social Lives of Male African Elephants
male African elephants turn out to have surprisingly rich social lives
By Caitlin O'Connell edited by Kate Wong
Young male elephant (right) greets an older male (left)
I woke up in the research tower to a soft exhale and then a trickle of water at the waterhole below
The full moon hung low and dim in the west
I wondered which of my elephant study subjects had come to drink at this hour
and they typically don’t show up here at the Mushara waterhole in Namibia’s Etosha National Park until at least 11 A.M
But the full moon does funny things to elephant movements
I rolled over in my sleeping bag to look out at the visitors
Two bulls stood at the source of the spring
and the younger one had his back toward me
Focusing my binoculars on the edges of their ears
I could see a telltale tab on the left ear of the older one and two sharp cuts on the right ear of the younger one
My team and I had been following the older bull
Abe hadn’t visited Mushara for the entire 2024 field season
his tusks having grown out nicely after breaking a few years ago
not only because I enjoyed observing his endearing patience with younger males but also because he was one of my most important study subjects
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Abe was an integral member of a social group whose behavior I had been focused on documenting for years
African elephants have matriarchal societies
in which elder females lead families made up of their female relatives and young to water
Most of what we know about elephant social behavior has come from studies of these female-led family groups
leave their family group as they reach adolescence and go off to live as bachelors
little research was conducted on the social dynamics of adult male elephants because they were thought to be largely solitary
It seemed safe to assume their social lives were simple
with none of the complex rules and rituals seen in the family groups
When young male elephants visit the waterhole with their families
they get to interact with bachelor males and learn who they can trust when the time comes to set out on their own
But research that I and other scientists have been carrying out is finally starting to fill this gap in our understanding of elephant behavior
Our long-term observations of Abe and the other males who have visited Mushara over the past two decades reveal that male elephant societies have incredibly intricate rules
are sensitive social animals that crave companionship and need emotional support
Their experiences of growing up parallel those of human males
who undergo similar social changes and pressures as they enter the adult world—from backslapping in the bar
to defending another’s honor by taking on a bully
Having a social network of other trusted males may be important for navigating the transition to adulthood
These findings indicate that this highly intelligent species is even more socially complex than we previously understood it to be—and could inform vital efforts to manage and protect these animals in the wild and in captivity
One of our most exciting discoveries concerns vocal communication
Researchers have long known that female African elephants use vocalizations to coordinate group behavior
In family groups gathered at a water source
align herself toward the direction she wants to leave in and then rumble a call to action—“Let’s go!”
It’s understandable why families would engage in this behavior
Coordinating a group of 20 to 40 or more elephants isn’t a simple task
Using a vocal tool to call the group into line is well documented in other species
one ape’s grunt triggers another to grunt
until consensus is reached and all leave the area together
who sneeze to reach consensus about departure
engaged in similar vocal coordination was unclear
I suspected based on a handful of observations that the male elephants were indeed communicating in this way
but I needed a larger sample to prove the pattern was real
The male elephants that congregate at Mushara tend to be most vocal at night
which also happens to be when conditions are best for recording their sounds
And so I found myself working the late shift during our field seasons
timing my observation sessions to when the bachelors were chattiest
Recently my collaborators and I published a paper containing the fruits of this labor
We were able to show that when a group of male associates leaves the waterhole
one of the core members of the group flaps his ears gently while emitting a long
low rumble—the “let’s go” signal
the others respond with their own rumble while following the instigator away from the water
each waiting for another to finish their call before contributing their own say in the matter
The paper marks the first time such coordination has been documented in male elephants
as well as the first time vocalizations have been shown to trigger action within male groups
When I first witnessed groups of bonded males engaging in this kind of vocal ritual
Why would these males want to leave together
in which the dominant females who determine the group’s movements are acting in the interests of their close relatives
male elephants have traditionally dispersed far from their natal group
This arrangement meant that they were often not related to the other males with whom they associated
Bonded males arrive at the waterhole together using vocal coordination (top)
Such groups also coordinate their departure this way
After a socially connected male initiates the "let's go" rumble
the rest of the group members take turns rumbling their assent as they walk away together (bottom)
Leaving together implies they’ll be sharing resources at their next destination
I had seen powerful older males shake full-grown acacia trees to make the seed pods—a favorite food of elephants—shower down to the ground
On some of these occasions a younger bull or two came in to sample the spoils
Usually established group members would shove chancers away from the prized pods
but they tolerated ones who had arrived at the invitation of the older male
There had to be some advantage to older males inviting younger males to join them in their travels
Why would older males want to share with younger associates when they didn’t have to
What advantage was there in staying together
Could these males want the companionship even if it meant sharing
Did male elephants form even tighter-knit groups than previously thought
Our observations of vocal coordination among the Mushara males sparked many such questions
Studies of other aspects of their behavior have led to some answers
the most obvious ritual exchange between male associates is what we call the greeting
A younger individual often initiates the greeting
placing the tip of his trunk either within or just at the entrance to the mouth of a buddy or an older male
The greeting is typically reciprocated—unless the recipient is very high-ranking
the gesture is more like a salute of one or more individuals to a general or a kiss of a royal’s ring
the behind-the-scenes decorum in male engagement is so fine-tuned it’s like the list of steps for having tea with the queen
We can look at the ritual of play for another example
play is important for building trust and developing skills needed to compete for mates later in life
Our data show that individuals of all ages within a bonded group of male elephants solicit play with other individuals in equal amounts
younger elephants must wait until the older one initiates the play bout
Such conventions are hallmarks of a very sophisticated society
Each ritual helps to reinforce the rules of engagement within the group
This is all very much in keeping with what we already know about elephant intelligence
empathy and sense of self based on studies of the female-led family groups
What’s new and surprising here is that we’re documenting these group dynamics in males
who were long thought to be mostly solitary
And not only do we see rituals in these male groups
but the groups also engage in behaviors indicative of a hierarchical social structure
In our study of vocal coordination triggering group action
a case of active leadership in male elephants
An active leader is one who solicits others to follow him and exerts social influence through means of dominance ranks
passive leadership might occur when an older individual is followed by an uninvited youngster
the older individual often signals his displeasure by turning around and shaking his head
This response usually deters unwanted companions
but it depends on how determined the younger one is
My colleagues and I got our first inkling of male leadership in the Mushara social club back in 2004
A dominant bull we named Greg seemed to call the shots
Long conga lines of males of all ages and sizes followed Greg in and out of the waterhole
He was beloved by his underlings and either feared or revered by many of his contemporaries and elders
He led with a carrot-and-stick style—forgiving of the transgressions of youngsters but hard on his contemporaries
Bachelor males of all ages benefit from socializing with other males
Such interactions can lead to lifelong companionship
When Greg decided it was time to depart Mushara
turn in the direction he planned to go and invite the others to leave with a “let’s go” rumble
ritualized rumbles from the rest of the group
On occasion Greg would give some of the youngest ones a gentle shove to expedite any deliberations
The simple act of inviting others to leave with him showed that Greg and males like him
actually make an effort to keep the group together
This behavior is news because leadership in male social groups is not a given in mammals
A dominant male baboon has seemingly no interest in leading his underlings
His primary interest is in securing the best resources for himself
It’s the female baboons who do the leading
as I observed Greg and the other males at the Mushara
I thought about why male elephants might come together in these groups and coordinate their actions
Forming coalitions could be one explanation
Ritualized vocal coordination could solidify and reinforce bonds the elephants needed to work together to intimidate or push off rival males
An unwanted individual could suddenly find himself facing a wall of imposing elephant ears
It would be a lot easier to gain access to the best resources as a group than for an individual to defend them on his own
And considering that the Mushara waterhole was one of only three water points in the region
I couldn’t help wondering whether there were other factors in play
Perhaps there was a reproductive advantage for older males who adopted the emotionally needy underlings
when elephants were free to roam in an environment unobstructed by fences built by humans
male elephants left their natal families during their coming-of-age testosterone spikes and were thought to settle far from their genetic line
This dispersal pattern maintained the diversity of the elephant gene pool
and it implied that males found socializing together were not related
But the realities of modern Africa are such that elephant movements are often impeded by human activity
and young males can’t move as far away from their families as they might have in the past
Older male Abe (front) and younger male (back) listen for the arrival of their companions
This ecological shift increases the odds that a bachelor will encounter his male relatives on the landscape after dispersing from his natal group
If males within bonded groups were in fact related to one another
their genetic line would have a better chance of getting passed along if older males could help ensure that their relatives survive and reproduce
can smell related individuals and hence avoid inbreeding
It occurred to me that male elephants might have a similarly innate ability to recognize relatives
a father elephant might be able to identify his offspring through olfactory cues and choose his companions accordingly
another recent study by my team indicates that male elephants track one another over space and time in a way that could be achieved only through olfaction and possibly vocalizations
We found evidence that males remotely monitor one another’s locations and move toward or away from other elephants depending on their social relationships
adult males find mates by listening for the distinctive calls of estrus females and smelling olfactory cues in their urine.)
We are still studying the relatedness of individuals in these bachelor groups
Future findings may shift our understanding of whether
males are acting to maximize their ability to reproduce and pass their genes to the next generation
There may be additional factors to consider as well
Another advantage of keeping a group together is companionship
who are still struggling to adjust socially to life outside a huge family
certainly benefit from older males who can model good behavior
Older males may gain from this association
In another recent study we conducted on male elephant character
we showed that adult males profit psychologically from the companionship of younger males
We collected behavioral data from 34 individually known males at Mushara over a five-year period and found that the presence of young males in a group reduces aggression and increases affiliative
Male elephants of all ages need socialization just like any other social animal
They seek interaction and bonding from youngsters
contemporaries and elders they can look up to
Given how important bonded groups are to elephants
I never would have expected that the structure of male elephant society was vulnerable to collapse
But after our 2011 fieldwork season at Mushara
Instead we witnessed a complete destabilization of the social club
Greg’s absence ushered in an era of small factions; no one took up his leadership torch
Abe just didn’t seem to have the titanium stomach required to rule
Being at the top is a lot of work—always having to posture
appeasing the youngsters while signaling dominance to one’s ever-watchful upwardly mobile contemporaries
facing the threat of being toppled by a potentially mortal blow at any moment
Yet somehow Greg was able to do it with surprisingly little stress
which we assessed by collecting and analyzing his feces over the years
now displays signs of active leadership and may one day become the leader of the social club at Mushara
Some 14 years after Greg’s disappearance
Young adults Kelly and Shaka have shown signs of wanting to mentor the next generation
They have allowed younger males to explore their faces and tusks
place tentative trunks in their mouths and hang a trunk on their tusk
They have even permitted two youngsters to hang their trunks on their tusks
I’d give anything to know what these young males are thinking in these moments of exploration and seeming admiration of the grown-ups
Puberty is a traumatic period of uncertainty for these youngsters
For a male that has grown up within a tight-knit elephant family
it’s a big social adjustment to live outside the family
I suspect that any ritual of connection would give a young male a sense of belonging—whether it be a secret handshake or tolerance from an elder
Acceptance goes a long way toward easing the transition to adulthood
It takes a certain kind of male character to have patience with these youngsters—and to take up the mantle of leadership
the right stuff seems to be a combination of self-confidence and a sense of decorum that gains the confidence of the group
Leaders tend to be the most socially integrated members of the group
After an older male allows young males to explore his face and body with their inquisitive trunks
the next sign that he has an interest in mentorship is the act of inviting these younger males to join him as he departs the waterhole
I can tell from the perky posture of the young males who receive such an invitation that they understand exactly what it means—that they have been accepted into the most important social club of their lives
I eagerly await these moments of active leadership
The more we understand about the complexities of male elephant society
the better we will be able to protect and manage this species
All our recent findings and the findings of others further speak to the social nature of male elephants
people who manage captive elephants might be able to improve their welfare by putting the males in mixed-age groups and organizing these groups around older males who have demonstrated leadership potential
efforts to manage elephant populations in the wild ought to factor in the importance of postreproductive bulls to coming-of-age males
Not only does their presence keep younger males
but they also contribute to the younger males’ sense of belonging
Now every time I hear a male “let’s go” rumble exchange
I jump out of my sleeping bag and turn on the sound recorder and the night vision scope
I quickly document who is involved in the exchange—who initiated and who responded and when
These observations give me hope for a return to the good old days at Mushara
All too soon after Abe and Andries finally showed up at the waterhole last year
and Earth’s largest land animal emitted the lowest-frequency vocalization on the planet
It was the “let’s go” rumble—the invitation to accompany him
It was the call that Abe had so often enjoyed responding to but rarely initiated himself
Andries answered the call to action with an equally long and low response
I rolled over and tucked myself back into my sleeping bag for just a little while longer
In an hour the camp would be a buzz of activity to pack up and leave
and I was grateful for Abe’s appearance
even if only in the final hours of my stay
It gave me peace of mind to know he was still out there
offering new coming-of-age males safe harbor
The sound of the “let’s go” duet faded
I imagined Abe and Andries saying to each other
I’ll go high!” as they harmonized and disappeared into the bush
bound for their next destination in good company
Why Animals Play. Caitlin O'Connell; August 2021. ScientificAmerican.com/archive
Caitlin O'Connell is a behavioral ecologist at Stanford University's Center for Conservation Biology and Harvard University's Center for the Environment
Her research focuses on elephant behavior and communication
O'Connell's latest book is Wild Rituals: 10 Lessons Animals Can Teach Us about Connection
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and they typically don’t show up here at the Mushara waterhole in Namibia’s Etosha National Park until at least 11 A.M
After a socially connected male initiates the \\"let's go\\" rumble
Caitlin O'Connell is a behavioral ecologist at Stanford University's Center for Conservation Biology and Harvard University's Center for the Environment
Why Animals Play. Caitlin O'Connell; August 2021. ScientificAmerican.com/archive
Whether you are part of our community or are interested in joining us
we welcome you to Washington University School of Medicine
Mouse study may lead to new approaches for contraception
by Mark Reynolds•May 2
a BJC Investigator and professor of cell biology and physiology
has shown in mice that sperm have a temperature-controlled switch that changes their movements and is key to male fertility
have evolved to keep male reproductive organs cooler than their core body temperature
In mammals — including us humans — sperm are surprisingly picky about temperature
thriving best in conditions a few degrees cooler than normal body temperature
given that the female reproductive tract is warmer than normal body temperature
how do these heat-sensitive swimmers manage to fertilize an egg in such a toasty environment
Research from Washington University School of Medicine in St
Louis has an answer: warmer temperatures — such as those of the female reproductive tract — trigger a specific signal that activates sperm
changing their behavior from the relatively smooth motions used for navigation to the thrashing and twisting motions needed to enter the egg for fertilization
The discovery of that trigger provides a possible target for male contraceptives and treatments for male infertility
the researchers showed that a specific protein common to all mammals triggers a hyperactive state in sperm when the surrounding temperature matches that of the female reproductive tract
published in Nature Communications April 17
also help explain the evolution of mammalian anatomy
“That hyperactive state in sperm is key for successful fertilization, and no one knew exactly how temperature triggers it,” said Polina Lishko, PhD
a BJC Investigator and professor of cell biology and physiology at WashU Medicine
“Our work has identified a temperature-controlled switch in sperm that triggers these increased movements precisely when they are needed during fertilization.”
All mammals share a particular protein embedded in the surface of sperm called CatSper
It controls the entry of particles needed to power the hyperactive movements in the tail-like flagella that propel the sperm forward
Temperature activation of CatSper was not known
however it had been believed to be activated by a combination of the pH level in the female reproductive channel and
because most mammalian sperm do not respond to progesterone
so there had to be another controlling factor to flip the CatSper switch
Mammalian evolution has developed ingenious ways to keep male reproductive organs at or below 34 degrees Celsius (93.2 degrees Fahrenheit)
lower the temperature of blood destined for their internal testes by first passing it through their dorsal fin; elephants use a similar system that cools blood through their ears
create and store sperm in testicles outside of their body
Animals without these cooling adaptations for their male reproductive organs
Using micron-scaled tools and techniques originally developed to study brain cells
Lishko’s team observed the pattern of electric charges distinctive to CatSper’s activation in individual sperm cells
measuring clear spikes when the temperature surrounding the cell surpassed 38 degrees Celsius (100.4 degrees Fahrenheit)
the sperm’s behavior switched from the relatively smooth motions they use for navigation to the hyperactive movements needed for them to enter the egg for fertilization
Lishko said that understanding temperature’s role in fertility may help to improve male contraception and treatments for infertility
targeting it would not affect other bodily functions
There have been attempts to develop contraceptives that deactivate the channel
but Lishko said those have not been very effective so far
The insights from this discovery might point to new approaches
it might be possible to activate CatSper with temperature thus
prematurely switching on this channel to drain the sperm of energy
so that by the time the sperm cell is ready to do its job and enter the egg cell
Swain DK, Vergara C, Castro-Arnau J, Lishko PV. The essential calcium channel of sperm CatSper is temperature-gated. Nature Communications. April 17, 2025. DOI: 10.1038/s41467-025-58824-0
This work was supported by the BJC Investigator fund
About Washington University School of Medicine
WashU Medicine has a storied history in MD/PhD training
recently dedicated $100 million to scholarships and curriculum renewal for its medical students
and is home to top-notch training programs in every medical subspecialty as well as physical therapy
j.church@wustl.edu
Mark Reynolds
r.mark@wustl.edu
and both occupational and physical therapy
specializing in science and medicine with publications in CNRS International
He is a former editor of McGill University’s Headway/En Tête research magazine and has won awards from the Canada Council for the Advancement of Education including for best science writing
He has a bachelor’s degree from Dalhousie University in Halifax
Could help determine which patients are likely to benefit from new Alzheimer’s drugs
GLP-1 medications tied to decreased risk of dementia
At WashU Medicine, we transform lives and shape the future of healthcare through pioneering research, world-class education, and unparalleled patient care. As one of the nation's largest academic clinical practices, we bring the full power of WashU Medicine to every patient, advancing treatment and training the medical leaders of tomorrow at Barnes-Jewish and St. Louis Children's hospitals
and more than 130 clinics across Missouri and Illinois
Consistently recognized among the nation's top institutions for research
we are driven to challenge convention and elevate care for all
“Is this creepy given I don’t know this person
or is it just me?” the woman asked on a community forum
Metrics details
Agonists and antagonists of the glucose-dependent insulinotropic polypeptide receptor (GIPR) enhance body weight loss induced by glucagon-like peptide-1 receptor (GLP-1R) agonism
while GIPR agonism decreases body weight and food intake in a GLP-1R-independent manner via GABAergic GIPR+ neurons
it remains unclear whether GIPR antagonism affects energy metabolism via a similar mechanism
Here we show that the body weight and food intake effects of GIPR antagonism are eliminated in mice with global loss of either Gipr or Glp-1r but are preserved in mice with loss of Gipr in either GABAergic neurons of the central nervous system or peripherin-expressing neurons of the peripheral nervous system
Single-nucleus RNA-sequencing shows opposing effects of GIPR agonism and antagonism in the dorsal vagal complex
GIPR antagonism and GLP-1R agonism both regulate genes implicated in synaptic plasticity
we show that GIPR agonism and antagonism decrease body weight via different mechanisms
these findings support the notion that both GIPR agonism and antagonism hold therapeutic value to accelerate GLP-1-induced weight loss
we find that the body weight and food intake inhibitory effects of GIPR antagonism are absent in global Glp-1r-deficient mice
suggesting dependency on GLP-1R-mediated signalling
single-nuclei RNA-sequencing (snRNA-seq) revealed that GIPR agonism and antagonism have opposing effects in the brain
with GIPR antagonism but not agonism mimicking the transcriptional responses of GLP-1R agonism in the dorsal vagal complex of the hindbrain (DVC)
and with GIPR antagonism and GLP-1R agonism both modulating DVC gene programmes implicated in synapse formation and neuronal plasticity
we show that while GIPR agonism and antagonism have similar effects on body weight and food intake
they do so via different neuronal mechanisms
depending on GLP-1R signalling to affect energy metabolism
Source data.
Source data.
Source data.
Body weight in grams (a) and percent (b) and food intake (c) of HFD-fed 14–16-week-old male C57BL6/J WT mice treated s.c
with a single dose (30 mg kg−1) of either a control mAb (vehicle; n = 5) or an anti-GIPR antagonist (ant.) antibody (n = 6)
Body weight in grams (d) and percent (e) and food intake (f) of HFD-fed 14–16-week-old male C57BL6/J global Gipr KO mice treated s.c
with a single dose (30 mg kg−1) of either a control mAb (vehicle) or an anti-GIPR antagonist antibody (n = 6 each group)
Body weight in grams (g) and percent (h) and food intake (i) of HFD-fed 14–16-week-old male C57BL6/J global Glp-1r KO mice treated s.c
with a single dose (30 mg kg−1) of either a control mAb (vehicle) or an anti-GIPR antibody (n = 6 each group)
Data in a–i were analysed by a two-way ANOVA with Bonferroni’s post hoc test for comparison of individual timepoints
Source data.
A schematic of the experimental design (a) and the body weight (b) of 36-week-old male C57BL/6J WT mice fed either a standard chow diet (cntrl.) or a HFD and treated s.c
with a single dose of either vehicle (vhcl)
an acylated peptide GIPR antagonist (1,500 nmol kg−1) or the GIPR:GLP-1R co-agonist MAR709 (50 nmol kg−1) (n = 6 each group)
UMAP representations of gene expression coloured by C12-level cell type in the DVC (c) and C25-level cell type in hypothalamus (d)
as well as by experimental group in the DVC (e) and hypothalamus (f)
A bar graph showing mean expression of Glp-1r and Gipr in the DVC and hypothalamus
The number of nuclei isolated from each brain region
The colours correspond to log-normalized expression values scaled to the maximum of each gene
Source data.
A comparison of log fold-change differences in gene expression in DVC neurons between male DIO C57BL/6J WT mice (DIO cntrl.) treated with acyl-GIP or the GIPR antagonist (ant.) (a)
GIPR antagonist versus MAR709 (e) or acyl-GLP-1 versus acyl-GIP (f) (n = 6 mice per group
from which n = 3 mice were pooled to receive n = 2 independent biological replicates per group)
UMAP representation of gene expression of DVC neurons coloured by neuron type (g)
and with expression of Glp-1r (h) and Gipr (i)
Heat maps showing mean gene expression of Glp-1r and Gipr in DVC neuronal populations (j) and experimental group (k)
and with the colour corresponding to log-normalized expression values scaled to the maximum of each gene
Source data.
Bar plots and UMAP representations of gene expression in DVC neurons of male DIO mice treated with a GIPR antagonist (ant.) (a)
acyl-GIP (c) or MAR709 (d) (n = 6 mice per group
The bar plots and UMAPs are coloured by Augur score
representing cell type-specific changes in gene expression of the treatment group relative to DIO vehicle controls
Volcano plots (log2 fold change (FC) versus adjusted P values from a two-sided Wilcoxon rank-sum test
corrected for multiple comparison) of differentially expressed genes (DEGs) following treatment with either the GIPR antagonist or acyl-GLP-1 in the top GIPR antagonist affected neuronal clusters Glut10 (e)
Venn diagrams show the overlap of significant DEGs (adjusted P < 0.05) from GIPR antagonist and acyl-GLP-1 groups
P values of DEGs were obtained by Wilcoxon rank-sum tests and were adjusted for multiple comparisons using the Benjamini–Hochberg method
Source data.
these data not only show that GIPR antagonism and GLP-1R agonism act on the same neuronal populations in the DVC
but also that they similarly downregulate gene programmes implicated in synaptic plasticity and synapse formation
UMAP representation of hypothalamic neuronal gene expression coloured by expression of Glp-1r (a)
experimental group (c) or C66-level neuron type (d)
Heat maps showing Glp-1r and Gipr mean gene expression in hypothalamic neuronal types
Bar plots and UMAP representations of gene expression in DVC neurons of DIO mice treated with a GIPR antagonist (f) or acyl-GLP-1 (g)
and UMAPs and bar plots are coloured by Augur score
representing cell-specific change in gene expression of the experimental group versus vehicle DIO controls (f and g)
The top ten most likely cell–cell communication events between DVC GABA4 or Glut 10 neurons and hypothalamic C66-19: Pomc.GLU-4; C66-46: Agrp.GABA-4 or all other hypothalamic neurons in DIO mice treated with vehicle
Cellphone P values are permutation-based P values
Source data.
these data suggest that GLP-1R agonism and GIPR antagonism may exert their effects on energy balance by downregulating signalling from DVC C35 GABA4 and C35 Glut10 neurons to hypothalamic feeding circuits
our snRNA-seq analysis revealed that GIPR antagonism
DVC neuronal gene expression changes correlate negatively in mice after treatment with GIPR agonism versus antagonism
but positively in mice treated with GIPR antagonism versus GLP-1R agonism
We observed the greatest transcriptional changes induced by GIPR antagonism in the C35 GABA4
which were also among the highest affected neuronal populations targeted by GLP-1R agonism
GIPR antagonism and GLP-1R agonism are separated from GIPR agonism in that they both similarly downregulate gene programmes indicative of neuronal plasticity and synapse formation
These findings further support the notion that GIPR antagonism and GLP-1R agonism are functionally related and act similarly on DVC neurons
and in a clearly distinct manner from GIPR agonism
we show here that GIPR agonism and antagonism affect body weight and food intake via different
with GIPR antagonism affecting body weight and food intake via modulation of GLP-1R signalling
The observation that gene expression changes induced by GIPR agonism versus its antagonism correlate negatively further argues that our GIPR agonist is not a functional antagonist
it seems plausible to hypothesize that weight loss induced by GIPR signal modification may involve modulation of the hypothalamic and/or hindbrain serotonergic system
Body composition was analysed using a magnetic resonance whole-body composition analyser (EchoMRI)
Plasma levels of glucose and insulin were measured after 6 h fasting
insulin (Humalog; Eli Lilly) was injected i.p
HbA1c was assessed from fresh blood using the DCA Vantage Analyzer (Siemens)
For assessment of glucose-induced insulin secretion
glucose was given orally at a dose of 4 g kg−1 in 6 h fasted mice
followed by blood sampling at timepoints 0
15 and 30 min after glucose administration
Commercially available enzyme-linked immunosorbent assays (ELISAs) were used according to the manufacturer’s instruction to measure insulin (Crystal Chem Zaandam
ab65336) or total cholesterol (Thermo Fisher Scientific
Brains were then fixed for 24 h at 4 °C in 4% PFA and then transferred to 15% sucrose for 24 h
tissues were extracted and fixed for 1–2 h in 4% PFA and transferred to 30% sucrose overnight at 4 °C
All tissues were frozen in Tissue-Tek O.C.T (Sakura Finetek
cut in 12–14 µm sections and placed on microscopic slides (Thermo Fisher Scientific
The slides were heated for 30 min at 60 °C followed by antigen retrieval using a steamer
then processed by the RNAscope Multiplex Fluorescent Reagent kit v2 (Advanced Cell Diagnostics
323270) according to the manufacturer’s instructions
a custom-made probe was designed to bind to the deleted exons of mouse Gipr (Advanced Cell Diagnostics
1138821-C1) and Vgat (Advanced Cell Diagnostics
amplifiers and dyes were added for visualization of GIPR and Vgat
The slides were incubated with rabbit anti-peripherin antibody (Thermo Fisher Scientific
PA316723; 1:200) for 1 h at room temperature
followed by 30 min incubation with goat anti-rabbit-HRP (Thermo Fisher Scientific
TSA vivid dyes 650 and 520 (Advanced Cell Diagnostics
both 1:500 dilution) were added to detect GIPR
peripherin or vesicular GABA transporter (VGAT)
Slides were counterstained with 4,6-diamidino-2-phenylindole (DAPI) (Advanced Cell Diagnostics
320858) and imaged using Leica SP8 Laser Confocal Microscope using LAS X (version 3.5.7.23225)
followed immediately by clamping of the bile duct and perfusion with collagenase P (Roche Diagnostics
Tissues were incubated in a 15 ml Falcon tube with 1 ml of collagenase P solution for 15 min at 37 °C
followed by the addition of 12 ml of cold G-solution (Sigma-Aldrich) and centrifugation at 586g at room temperature
The pellet was subsequently washed with 10 ml of G-solution (500 ml HBSS (Life Technologies
126615-25 ml) and 1% penicillin–streptomycin (Life Technologies
15140122)) and resuspended in 5.5 ml of gradient solution comprising 15% Optiprep (5 ml 10% RPMI (Life Technologies
11875093) + 3 ml of 40% Optiprep that was diluted from 60% Optiprep with G-solution (Sigma-Aldrich
and placed on top of 2.5 ml of the gradient solution
6 ml of the G-solution was added on the top
Samples were then incubated for 10 min at room temperature and centrifuged at 630g
The interphase was then collected and filtered through a 70 μm nylon filter (BD Falcon
Islets were handpicked by a micropipette under the microscope and cultured in RPMI 1640 medium (Life Technologies
Culture medium was removed and islet microtissues were equilibrated for 1 h with Krebs Ringer HEPES buffer (131 mM NaCl
1.2 mM MgSO4 and 2% BSA) containing 2.8 mM glucose
The supernatant was collected as a sample under the low glucose condition for 45 min incubation
and islets were incubated for another 45 min at 37 °C with Krebs Ringer HEPES buffer containing 16.7 mM glucose and supplements as above
The supernatant was collected as a sample under the high glucose condition and stored at −20 °C
native mouse GIP or GLP-1 (provided by Novo Nordisk) were diluted in 1× KRK buffer with 20 mM glucose to reach a concentration of 50 nM
Cells were subsequently treated with either mouse GIP or GLP-1 for 45 min
Insulin concentrations were determined using a Mouse Insulin ELISA (Crystal Chem
The hypothalamus and DVC were collected 8 h after drug administration and stored in liquid nitrogen
Mice were euthanized followed by immediate decapitation and then the skull was removed
An earlier alignment of the brain was determined using a brain matrix and the entire hypothalamus (includes all the nuclei) was collected by microdissection
The hindbrain DVC was microdissected in an area postrema-centric manner after removal of cerebellar cortex
Tissue samples were flash frozen into liquid nitrogen and frozen tissues were stored in liquid nitrogen vapour phase for further processing to single-nuclei isolation
Nuclei were isolated using the 10X Genomics Chromium Nuclei Isolation kit including RNase Inhibitor (10X Genomics
and using the 10X Genomics protocol for Single Cell Multiome ATAC + Gene Expression (10X Genomics
Nuclei concentration was determined using a Luna-II Automated Cell Counter (Logos biosystems
L40002) and adjusted to 6,250 nuclei per microlitre after pooling of n = 3 mice per sample
Nuclei were then processed using the 10X Genomics Chromium Next GEM Single Cell Multiome ATAC + Gene Expression (Rev
E) according to the manufacturer’s instructions
Pooled samples were loaded into two lanes per group for a total of 24 lanes across three 10X Chromium chips
Equal numbers of cells per sample were loaded on a 10X Genomics Chromium controller instrument to generate single-cell gel beads in emulsion at the Helmholtz Munich Genomics Core Facility
Single-nucleus multimodal libraries were sequenced using the Illumina NovaSeq 6000
FASTQ files were generated from base calls with bcl2fastq software v2.20 (Illumina)
Reads were mapped to the pre-built mm10 mouse reference (University of California Santa Cruz mm10 reference genome) using Cell Ranger ARC (v2.0.2
The resulting cell-by-peak and cell-by-gene matrices (ATAC and gene expression assays
respectively) from the 24 samples were integrated using Cellranger aggr (10X Genomics)
Then the expression of marker genes from the HypoMap was evaluated in each Leiden cluster
and then was manually annotated informed by marker gene expression and the majority cell type from scArches label transfer
The HypoMap hierarchical cell-type annotation framework was then used to map C25
as there is no comparable atlas and annotation framework to the hypomap for the DVC
each Leiden cluster was manually annotated into 35 fine-grained cell types (C35 cell type)
and were then mapped to the coarser-grained C12 and C2 levels of cell type based on the expression of marker genes
DVC neurons were further subdivided into individual clusters labelled by major neurotransmitter expression
drugs were aliquoted by a lead scientist in number-coded vials and most
handling investigators were blinded to the treatment condition
Analyses of glucose and insulin tolerance were performed by experienced research assistants who did not know prior treatment conditions
Further information on research design is available in the Nature Portfolio Reporting Summary linked to this article
Continued treatment with tirzepatide for maintenance of weight reduction in adults with obesity: the SURMOUNT-4 randomized clinical trial
Tirzepatide once weekly for the treatment of obesity in people with type 2 diabetes (SURMOUNT-2): a double-blind
Tirzepatide once weekly for the treatment of obesity
Tirzepatide after intensive lifestyle intervention in adults with overweight or obesity: the SURMOUNT-3 phase 3 trial
Effect of subcutaneous tirzepatide vs placebo added to titrated insulin glargine on glycemic control in patients with type 2 diabetes: the SURPASS-5 randomized clinical Trial
Tirzepatide versus insulin glargine in type 2 diabetes and increased cardiovascular risk (SURPASS-4): a randomised
Tirzepatide versus semaglutide once weekly in patients with type 2 diabetes
Once-weekly tirzepatide versus once-daily insulin degludec as add-on to metformin with or without SGLT2 inhibitors in patients with type 2 diabetes (SURPASS-3): a randomised
Tirzepatide vs insulin lispro added to basal insulin in type 2 diabetes: the SURPASS-6 randomized clinical trial
Efficacy and safety of a novel dual GIP and GLP-1 receptor agonist tirzepatide in patients with type 2 diabetes (SURPASS-1): a double-blind
Glucose-dependent insulinotropic polypeptide (gastric inhibitory polypeptide; GIP)
Therapeutic potential for GIP receptor agonists and antagonists
Loss of GIPR in LEPR cells impairs glucose control by GIP and GIP:GLP-1 co-agonism without affecting body weight and food intake in mice
Glucose-dependent insulinotropic polypeptide regulates body weight and food intake via GABAergic neurons in mice
The glucose-dependent insulinotropic polypeptide (GIP) regulates body weight and food intake via CNS–GIPR signaling
a novel dual GIP and GLP-1 receptor agonist for the treatment of type 2 diabetes mellitus: from discovery to clinical proof of concept
Unimolecular dual incretins maximize metabolic benefits in rodents
Optimized GIP analogs promote body weight lowering in mice through GIPR agonism not antagonism
Specific loss of GIPR signaling in GABAergic neurons enhances GLP-1R agonist-induced body weight loss
Hypothalamic and brainstem glucose-dependent insulinotropic polypeptide receptor neurons employ distinct mechanisms to affect feeding
Glucose-dependent insulinotropic polypeptide receptor-expressing cells in the hypothalamus regulate food intake
Discovery of a potent GIPR peptide antagonist that is effective in rodent and human systems
Anti-obesity effects of GIPR antagonists alone and in combination with GLP-1R agonists in preclinical models
Chronic glucose-dependent insulinotropic polypeptide receptor (GIPR) agonism desensitizes adipocyte GIPR activity mimicking functional GIPR antagonism
GIPR antagonist antibodies conjugated to GLP-1 peptide are bispecific molecules that decrease weight in obese mice and monkeys
A GIPR antagonist conjugated to GLP-1 analogues promotes weight loss with improved metabolic parameters in preclinical and phase 1 settings
Gut-derived GIP activates central Rap1 to impair neural leptin sensitivity during overnutrition
leads to enhanced weight loss and metabolic improvements in cynomolgus monkeys
GIP as a therapeutic target in diabetes and obesity: insight from incretin co-agonists
Sensory and motor physiological functions are impaired in gastric inhibitory polypeptide receptor-deficient mice
Glucose-dependent insulinotropic polypeptide (GIP) and its receptor (GIPR): cellular localization
Relevance of sympathetic nervous system activation in obesity and metabolic syndrome
GIP-induced vasodilation in human adipose tissue involves capillary recruitment
The gluco- and liporegulatory and vasodilatory effects of glucose-dependent insulinotropic polypeptide (GIP) are abolished by an antagonist of the human GIP receptor
TCF1 links GIPR signaling to the control of beta cell function and survival
Inactivation of the glucose-dependent insulinotropic polypeptide receptor improves outcomes following experimental myocardial infarction
Murine peripherin gene sequences direct Cre recombinase expression to peripheral neurons in transgenic mice
A new neuronal intermediate filament protein
NF-L and peripherin immunoreactivities define distinct classes of rat sensory ganglion cells
Neuronal GLP1R mediates liraglutide’s anorectic but not glucose-lowering effect
Spatiotemporal GLP-1 and GIP receptor signaling and trafficking/recycling dynamics induced by selected receptor mono- and dual-agonists
Cell type prioritization in single-cell data
Neuregulin-ERBB signaling in the nervous system and neuropsychiatric diseases
NGL-2 regulates input-specific synapse development in CA1 pyramidal neurons
NGL-1/LRRC4C deletion moderately suppresses hippocampal excitatory synapse development and function in an input-independent manner
The synaptic and neuronal functions of the X-linked intellectual disability protein interleukin-1 receptor accessory protein like 1 (IL1RAPL1)
HypoMap-a unified single-cell gene expression atlas of the murine hypothalamus
LIANA+ provides an all-in-one framework for cell–cell communication inference
CellPhoneDB: inferring cell–cell communication from combined expression of multi-subunit ligand-receptor complexes
Inferring neuron–neuron communications from single-cell transcriptomics through NeuronChat
Tanycytes control hypothalamic liraglutide uptake and its anti-obesity actions
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A genetic map of the mouse dorsal vagal complex and its role in obesity
Deep generative modeling for single-cell transcriptomics
Hierarchical progressive learning of cell identities in single-cell data
The arcuate nucleus mediates GLP-1 receptor agonist liraglutide-dependent weight loss
The hypothalamic glucagon-like peptide 1 receptor is sufficient but not necessary for the regulation of energy balance and glucose homeostasis in mice
Semaglutide lowers body weight in rodents via distributed neural pathways
Dissociable hindbrain GLP1R circuits for satiety and aversion
Liu, C. M. et al. GIPR-Ab/GLP-1 peptide–antibody conjugate requires brain GIPR and GLP-1R for additive weight loss in obese mice. Nat. Metab. https://doi.org/10.1038/s42255-025-01295-w (2025)
5-HT recruits distinct neurocircuits to inhibit hunger-driven and non-hunger-driven feeding
5-HT2CRs expressed by pro-opiomelanocortin neurons regulate energy homeostasis
Obesity medication lorcaserin activates brainstem GLP-1 neurons to reduce food intake and augments GLP-1 receptor agonist induced appetite suppression
Monomeric GLP-1/GIP/glucagon triagonism corrects obesity
Validation of HOMA-IR in a model of insulin-resistance induced by a high-fat diet in Wistar rats
Comparison between surrogate indexes of insulin sensitivity and resistance and hyperinsulinemic euglycemic clamp estimates in mice
Surrogate measures of insulin resistance: of rats
Comparison between surrogate indexes of insulin sensitivity/resistance and hyperinsulinemic euglycemic clamp estimates in rats
Revisiting energy expenditure: how to correct mouse metabolic rate for body mass
A guide to analysis of mouse energy metabolism
Scrublet: computational identification of cell doublets in single-cell transcriptomic data
SCANPY: large-scale single-cell gene expression data analysis
A step-by-step workflow for low-level analysis of single-cell RNA-seq data with Bioconductor
Mapping single-cell data to reference atlases by transfer learning
Heumos, L. et al. Pertpy: an end-to-end framework for perturbation analysis. Preprint at bioRxiv https://doi.org/10.1101/2024.08.04.606516 (2024)
Download references
This work was funded by the European Union within the scope of the European Research Council ERC-CoG Trusted no
The views and opinions expressed are however those of the author(s) only and do not necessarily reflect those of the European Union or the European Research Council
Neither the European Union nor the awarding authority can be held responsible for them
further received funding from the German Research Foundation (grant nos
SFB1123 and GRK 2816/1) and the German Center for Diabetes Research (DZD e.V.)
The skilful technical support of the Core Facility Genomics at Helmholtz Munich is highly acknowledged
De la Rosa Velazquez for their help with the snRNA-seq
Open access funding provided by Helmholtz Zentrum München - Deutsches Forschungszentrum für Gesundheit und Umwelt (GmbH)
These authors contributed equally: Robert M
Arkadiusz Liskiewicz & Daniela Liskiewicz
Institute of Experimental and Clinical Pharmacology and Pharmacogenomics
Obesity and Complications Therapeutic Area
Walther-Straub Institute for Pharmacology and Toxicology
designed and performed experiments and analysed and interpreted data
participated in the molecular design and interpretation of data
analysis of data and interpretation of results
supervised experiments and analysed and interpreted data
is a member of the scientific advisory board of ERX Pharmaceuticals
He was a member of the Research Cluster Advisory Panel (ReCAP) of the Novo Nordisk Foundation between 2017 and 2019
He attended a scientific advisory board meeting of the Novo Nordisk Foundation Center for Basic Metabolic Research
He received funding for his research projects by Novo Nordisk (2016–2020) and Sanofi-Aventis (2012–2019)
He was a consultant for Bionorica SE (2013–2017)
As former Director of the Helmholtz Diabetes Center and the Institute for Diabetes and Obesity at Helmholtz Zentrum München (2011–2018)
he has been responsible for collaborations with a multitude of companies and institutions
he discussed potential projects with and has signed/signs contracts for his institute(s) and for the staff for research funding and/or collaborations with industry and academia
including but not limited to pharmaceutical corporations like Boehringer Ingelheim
he was/is further responsible for commercial technology transfer activities of his institute(s)
including diabetes related patent portfolios of Helmholtz Zentrum München as
none of the above funding sources were involved in the preparation of this paper
receives research funding by Novo Nordisk and has received speaking fees from Novo Nordisk
is a co-inventor on intellectual property owned by Indiana University and licensed to Novo Nordisk
The other authors declare no competing interests
Nature Metabolism thanks Alice Adriaenssens
reviewer(s) for their contribution to the peer review of this work
Primary Handling Editor: Christoph Schmitt
in collaboration with the Nature Metabolism editorial team
RNAscope validation of GIPR deletion in 20 week-old male chow-fed Vgat cre+ Giprwt/wt (WT) and Vgat cre+ Giprflx/flx (KO) mice (pictures are representative examples of n = 3 mice per group) (a,b)
Absolute values of body weight development (c–e) and total change in body weight (f) of 33-wk old male C57BL/6 J wildtype (WT) or Vgat-Gipr knockout (KO) mice treated daily over 24 days with either vehicle
or the combination of acyl-GLP-1 (10 nmol/kg) and a GIPR antagonist (1,500 nmol/kg) (n = 8 each group)
Absolute changes of body composition of 36-wk old male C57BL/6 J wildtype and Vgat-Gipr KO mice treated either with Vehicle (n = 8 WT and n = 8 KO)
or the co-therapy of acyl-GLP-1 and the GIPR antagonist (n = 8 WT and n = 7 KO) (g,h)
Data in panel d and e were analyzed by repeated measures 2-way ANOVA with Bonferroni’s post-hoc test for comparison of individual time points
Data in panel f-h were analyzed using 1-way ANOVA
Data represent mean ± SEM; asterisks indicate * p < 0.05; ** p < 0.01 and *** p < 0.001
Individual p-values are shown in the Data Source file
Source data
Expression of peripherin corrected by the housekeeping gene peptidylprolyl isomerase A (Ppia) in hippocampus (n = 6)
dorsal root ganglion (n = 5) and iWAT (n = 4) of 15-week old male chow-fed Per-Cre+Giprwt/wt mice
and in pancreas (n = 7) of 45-week old male chow-fed Per-Cre+Giprwt/wt mice (a)
Expression of peripherin corrected by the housekeeping gene Ppia in the cerebellum (n = 8)
colon (n = 8) and trigeminal ganglion (n = 7) of 12-week old male chow-fed WT mice (b)
Expression of Gipr in dorsal root ganglion (n = 9 WT
eWAT (n = 12 each group) (g) and pancreas (n = 12 each group) (h) of 45-week old male mice Chow-fed Per-Cre+Giprwt/wt (WT) and Per-Cre+Giprflx/flx (KO) mice
Expression of Gipr in the cerebellum (n = 8 each group) (i)
n = 8 KO) (o) and colon (n = 8 each group) (p) of 12-week old male mice Chow-fed WT and KO mice
RNAscope validation of Gipr deletion in trigeminal ganglion of 44-week old male chow-fed WT and KO mice (q) and of the DRG of 51-week old male chow-fed WT and KO mice (r)
Data in panels q and r are representative examples of n = 3 mice each group
Data in panel c-f and h-p were analyzed using two-sided
data in panel g were analyzed two-sided by Mann-Whitney test
Source data
Body weight development of female C57BL6/J Per-Cre+Giprwt/wt (WT) and Per-Cre+Giprflx/flx (KO) mice fed with a HFD (n = 8 each group) (a)
Fat and lean tissue mass of 35-week old female WT and KO mice (n = 8 each group) (b,c)
Cumulative food intake of 52-week old female WT (n = 6) and KO mice (n = 7) (d)
locomotor activity (f) and respiratory exchange ratio (RER) (g) of 52-week old female WT (n = 6) and KO mice (n = 7)
Glucose tolerance (h) and corresponding area under curve (AUC) (i) after i.p
dosing with 2 g/kg glucose in 48-week old female WT and KO mice (n = 7 each group)
Glucose-induced insulin secretion (j) and corresponding AUC (k) after oral glucose bolus administration of 4 g/kg glucose in 54-week old female WT and KO mice (n = 6 each group)
dosing with 0.75U/kg insulin (Humalog) in 50-week old female WT (n = 6) and KO mice (n = 8)
Fasting levels of blood glucose (n = 8 each group) (m) and insulin (n = 6 WT
n = 7 KO) (n) in 54-week old female WT and KO mice
as well as fasting plasma levels of triglycerides (n = 6 WT
n = 7 KO) (p) in 55-week old female WT and KO mice
Data in panel a,c,f and g were analyzed by repeated measures 2-way ANOVA with Bonferroni’s post-hoc test for comparison of individual time points
Data in panel a,d,e,h,j and l were analyzed using 2-way ANOVA and with Bonferroni post-hoc comparison of individual time points
Data in panel e was analyzed using ANCOVA with body weight as covariate
Data in panel b,c,i,k,m-p were analyzed using Student’s two-tailed
Source data
Body weight development of 47-week old male C57BL/6 J wildtype (WT) and Per-Gipr knockout (KO) mice treated daily with either vehicle
or the combination of acyl-GLP-1 (10 nmol/kg) and a GIPR antagonist (1,500 nmol/kg) (n = 8 each group) (a)
Body composition (n = 8 each group) 47-wk old male C57BL/6 J DIO wildtype and Per-Gipr KO mice after 25 days of treatment (b,c)
Data in panel a was analyzed by 2-way ANOVA with Bonferroni’s post-hoc test for comparison of individual time points
Data in panel b,c were were analyzed using 1-way ANOVA
Source data
Comparison of Log fold change differences in gene expression between male DIO C57BL/6 J wildtype mice treated with acyl-GIP or the GIPR antagonist (a)
the GIPR antagonist (f) (n = 6 mice per group
Bar plots are ranked Augur score in mice treated with either acyl-GIP (g) or MAR709 (h)
representing cell-specific change in gene expression of the respective groups relative to Vehicle-treated DIO controls
The top 10 most likely cell-cell communication events between DVC GABA4 or Glut 10 neurons and hypothalamic C66-19: Pomc.GLU-4
or all other hypothalamic neurons in mice treated with MAR709 (i)
Cellphone p-values are permutation-based p-values
Lr means are mean ligand-receptor expression
Source data
UMAP representations of gene expression of DVC non-neuronal cells colored by experimental group (a)
or expression of either Glp-1r (c) or Gipr (d)
Bar plots with Augur scores of DVC non-neuronal cells from mice treated with either the GIPR antagonist (e)
Bar plots are ranked and colored by Augur score
representing cell-specific change in gene expression of the respective group relative to vehicle-treated DIO controls
Source data
UMAP representations of gene expression of hypothalamus non-neuronal cells colored by experimental group (a)
(e) Heatmap showing Glp-1r and Gipr mean gene expression in hypothalamic non-neuronal cell types
Color corresponds to log-normalized expression values scaled to the maximum of each gene
Bar plots showing Augur scores of hypothalamic non-neuronal cells from mice treated with the GIPR antagonist (f)
Source data
and by the experimental group from each study (e)
Source data
Pairwise heatmap showing the proportion of cells labeled by Hes et al
predicted to belong to each DVC cell-type from this study (x-axis) using scHPL
Source data
Pairwise heatmap showing the proportion of cells labeled by Ludwig et al
Source data
RNAscope analysis of GIPR in the nodose ganglion of 59-week-old male chow-fed WT and Per-GIPR KO mice
Data are representative examples of n = 3 mice each group
Statistical source data for Extended Data Fig
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DOI: https://doi.org/10.1038/s42255-025-01294-x
After finishing the indoor season as a second team all-american Lewis looked to continue his success into the outdoor season
The 2024 OVC Male Field Athlete of the Year would have no problem picking up right where he left off securing a first place finish in shot put and two second place finishes in hammer throw and discus at the Georgia Tech Invitational
Coach Crumpecker spoke on Lewis and his transition from indoor to outdoor season
“He's coming off of a tremendous indoor season
and he's continuing that into the outdoor season
and I expect that you'll continue to see bigger and bigger things as we get closer to the end of this (season).” Crumpecker said
Lewis was honored to be rewarded for his hard work and looks to continue his success
“It feels good to be recognized by the OVC especially for
the hard work I put in over the fall and this whole season.” Lewis said
or at least one more before the end of the season.”
While Lewis has shown exceptional strength throughout his tenure at SEMO (Southeast Missouri State University) his teammates say it’s his technique that has allowed him to thrive
Sophomore first year thrower Jachys Brown spoke on what puts Lewis ahead of the competition
he warms up the way he’s supposed to and he just studies everything…I feel like that’s what separates him.” Brown said
“I really expect him to get to Nationals and do great
I feel like he could be a good contender at Nationals I really do.”
Lewis along with the rest of the Redhawks will look to have another good outing as they will be back in action at the Memphis Tiger Invitational on May 2
SPLC and Florida Decides Healthcare File Lawsuit Challenging DeSantis’ Attack on Direct Democracy — Read More
Far-right extremists celebrated President Donald Trump’s 2024 election as an opportunity to further entrench a male supremacist culture that is permissive to violence against women
Through his appointments of white men dogged by allegations of sexual harassment
Trump has publicly affirmed his hostility for feminism to his male supporters and fed the male supremacist belief that white men are victims threatened by a changing social order
whose privilege must be restored and protected
There are serious consequences to selecting male supremacists and potential perpetrators of violence against women to top government positions
both for the women working in these agencies and for the rest of the country
His actions will strengthen hierarchies of both gender and race
As the election results came in last year on Nov. 5, white nationalist Nick Fuentes tweeted: “Your body, my choice. Forever.” The tweet was viewed over 100 million times and became a popular refrain to harass girls in school
a social media influencer who faces allegations of rape and human trafficking
“You no longer have rights.” A Texas Proud Boys chapter shared a meme depicting The Handmaid’s Tale dystopia
see Trump’s victory as a rebuke of feminism and progressive politics
He symbolizes the forceful rollback of women’s rights
Trump’s new administration has welcomed many personnel with histories of sexual misconduct. In the first few weeks after he was reelected to the presidency, Fortune reported nearly one-third of his Cabinet nominees and other key members of his administration had allegations of sexual misconduct
a self-identified “raging misogynist” who was also accused of domestic violence and subject to a temporary restraining order in 2022
which was dismissed days later as part of an agreement
as general counsel for the Office of Personnel Management
white men — as the ultimate victims of a changing social order
In their article for the Council on Foreign Relations
Jacob Ware and Noël James wrote that by nominating candidates “who have been accused or found guilty of sexual harassment for consideration for top positions leading the government
a message is sent to the public that renders these crimes as insignificant in regards to service in the government
alongside broader approval that legitimizes crimes related to violence against women.”
who has been accused of sexually harassing and groping his former babysitter
now leads the Department of Health and Human Services — an agency where he has a great deal of power over policies that impact women’s health and
the health of women living in poverty and women of color
On a podcast in November 2024, Hegseth said
“I’m straight up just saying we should not have women in combat roles.” In his 2024 book The War on Warriors: Behind the Betrayal of the Men Who Keep Us Free
“Women cannot physically meet the same standards as men.” In another comment in this book
But not in the military.” Hegseth’s actions as secretary of defense will have a disproportionate impact on Black women
who make up 29% of active-duty enlisted women
Male supremacists celebrated these views and what they indicated about women’s position in society
which the Southern Poverty Law Center defines as a hate group
“Will Donald Trump’s 2nd Term Turn The US Into A More Patriarchal Society?” The user specifically noted Hegseth’s views on women and his hope this would help diminish the influence of feminism
Despite some support for Trump and Hegseth in the thread
arguing that his views and policies are not extreme enough for their tastes
Title IX is part of federal civil rights legislation that bans sex discrimination at educational institutions that receive federal funding
This includes gender- and sex-based harassment
MRAs are ideologically opposed to these Title IX protections
which they believe unfairly privileges women at the expense of maligning men and contributing to what they consider to be “an epidemic of false rape allegations.”
Reporting from Democracy Forward revealed an extensive relationship between DeVos’ Department of Education and these organizations
who were directly involved in drafting new Title IX regulations
which weakened federal guidance on reporting
investigating and responding to sexual assault and harassment allegations at K-12 schools and universities while offering new protections to accused perpetrators
The new Trump administration’s decision to reaffirm this guidance leaves survivors vulnerable and unsupported
human trafficking and forming an organized crime group
Romanian authorities identified at least 35 known victims of their sex-trafficking operation
the Tate brothers continued this abusive operation even while under house arrest
and its recommendations threaten the privacy rights of patients who receive abortion care
These restrictions will have a disparate impact on women of color and those who live in poverty, in addition to further exacerbating the gap in maternal mortality between white and Black women
Despite trying to distance himself from Project 2025 during his campaign
Trump has since selected several of its authors to fill key positions within his administration and has taken steps to force this deeply unpopular policy agenda on the American people
The ideology of the Trump administration seems to be inching closer to that of organized far-right extremists
“You got to protect women from themselves … they’re indecisive
This is the version of protection the Trump administration is advocating for: “protecting” women by restoring traditional restrictive gender roles and enforcing a social hierarchy dominated by cisgender men and permissive of violence against women
President Trump symbolizes the forceful rollback of women’s rights
The SPLC and American University’s Polarization & Extremism Research & Innovation Lab have published Not Just a Joke: Understanding & Preventing Gender- & Sexuality-Based Bigotry
The guide helps readers identify warning signs that a young person has become susceptible to these ideologies
best practices for responding and ways to build resilience against manipulation
It also provides guidance on how to support and care for those who have been harmed by people who have been influenced by or hold bigoted beliefs
Comments or suggestions? Send them to [email protected]. Have tips about the far right? Please email: [email protected]. Have documents you want to share? Submit here
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The Southern Poverty Law Center400 Washington AvenueMontgomery
They were likely on your mind since April 25 was World Penguin Day
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A progressive Maine law firm that represents several school districts throughout the state issued a memo alongside the Maine School Board Association on Friday arguing that Maine school districts are required to continue forcing female athletes to compete against male athletes who claim they are transgender
The email, dated May 2 with the subject line “Title IX FAQ,” was sent to Maine school board chairs and members from MSBA President Faye Anderson, Executive Director Steven Bailey and Drummond Woodsum attorney Melissa Hewey
“This memo was developed in response to questions submitted to the Maine School Superintendents Association following an April 17th meeting with the Maine Department of Education,” the email reads
“The attached FAQs address concerns related to Title IX
and compliance with both state and federal law,” it reads
is to provide all Maine public school board members with “clarity and guidance as you navigate these important legal obligations.”
“We think it is especially important that all public school board members receive this memo,” the email reads
followed by a statement that Drummond Woodsum attorney Melissa Hewey made “major contributions” to the development of the FAQ document
Federal Funding and Maine Schools,” answers four questions regarding the legal precedence between federal and state law
and whether Maine schools must allow transgender athletes to participate in girls’ athletic events
The Maine School Board Association (MSBA) and the Maine School Superintendents Association (MSSA) are two nonprofit organizations that collectively form the Maine School Management Association (MSMA)
a taxpayer-funded quasi-governmental organization that promotes left-wing education policies
The memo comes amid an ongoing legal battle between the Trump administration and the State of Maine for Maine’s refusal to comply with Donald Trump’s executive order “Keeping Men out of Women’s Sports.”
The order established the current administration’s interpretation of Title IX anti-discrimination laws that considers it a violation for males to be allowed in women’s sports
It is argued in the Drummond Woodsum FAQ memo that “State and federal law take precedence over executive orders,” and that the Maine Human Rights Act
which prohibits discrimination based on gender identity
is “not preempted by Title IX,” and that Maine schools must comply with both laws
“The Maine Human Rights Act is currently consistent with Title IX,” it is claimed in the memo
The memo goes on to state that while Title IX does prohibit discrimination in education on the basis of sex
it “has nothing to do with athletics.”
In response to the question “Must Maine schools allow transgender girls to participate in girls’ athletic events?” the memo answers
“The Maine Human Rights Act requires this as do the rules of the Maine Principals’ Association and Title IX does not prohibit it,” the memo reads
“If state law is amended as has been proposed
The Maine Principals’ Association (MPA) is a private nonprofit corporation that regulates interscholastic high school athletics statewide
Last year, the MPA adopted a new “Gender Identity Participation Policy” to make it easier for transgender or non-binary students to participate on high school sports teams in accordance with their “gender identity.”
The final question addressed in the FAQ memo is whether Maine school administrative units (SAUs) that choose to align with President Trump’s executive order and keep males out of girls’ athletics events will receive federal funding
even if that funding is withheld from the state
This question appears to be in reference to Hodgdon school district (MSAD 70), whose board voted unanimously last month to adopt the federal government’s interpretation of Title IX
becoming the first school board in the state to do so
It was indicated by Attorney General Pam Bondi at a press conference last month announcing the Department of Justice’s lawsuit against Maine for their noncompliance with Title IX that MSAD 70 would not be losing out on federal funding withheld from Maine by the Trump administration
it is argued in the Drummond Woodsum memo that school districts who do decide to align with the Trump administration will indeed be affected by federal funds being withheld from the state at large
“The State of Maine is the grantee for virtually all federal funds flowing to schools in the state
so if one or more federal agencies decides to stop funding to the state
that decision will affect all SAUs regardless of their position on transgender participation in athletics,” the memo states
“There is no provision in the law for the federal government to provide funds directly to SAUs.”
The full MSBA/Drummond Woodsum memo to Maine school board members can be read below:
a school board member in Piscataquis county
was one of the Maine school board members who received the FAQ memo via email on Friday
“Today I received this email and attachment from the Maine School Boards Association with Title IX guidance for public schools
Namely: 1) state and federal law take precedence over executive orders; 2) the Maine Human Rights Act is not preempted by Title IX
therefore schools must comply with both laws; and 3) Maine schools must allow transgender girls to participate in girls’ athletic events because the MHRA requires it and Title IX does not prohibit it,” Perkins wrote in an email to the Maine Wire on Friday
Perkins pointed to Drummond Woodsum Attorney Hewey’s biography on the law firm’s website indicates that she has given past presentations entitled “Student Rights: Zooming in on the Rights of Transgender and Gender Expansive Students,” “The New Era of Title IX” and “Supporting Transgender Students in Maine Schools.”
“Knowing Melissa Hewey gave ‘major contributions’ to the document
leads me to believe that public school board members in Maine are receiving slanted advice and direction,” Perkins said
Friday’s memo is not the first time Drummond Woodsum has issued a memo to Maine public school officials related to obstructing the policies of the Trump administration
In January, the law firm sent out a memo advising public school officials on how they can effectively resist potential immigration enforcement actions that may involve the families of minors who attend government-run schools
including by destroying evidence of potential immigration crimes
Regarding information on noncitizen students in public schools
Drummond Woodsum advised their public school administrator clients
“[T]his information should not be stored as part of a students education record and should be destroyed as soon as it is no longer needed.”
Drummond Woodsum is the law firm representing the Great Salt Bay Community School in a lawsuit funded by Damariscotta area taxpayers
The lawsuit was initiated by a parent who uncovered that a conditionally licensed social worker at the school was covertly trying to persuade her 13-year-old daughter to embrace a gender transition
Edward Tomic is a reporter for The Maine Wire based in Southern Maine
Massachusetts and is a graduate of Boston University
He can be reached at tomic@themainewire.com
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Time to either say no or just go with it same with you parents
Your schools and politicians are against you
are being held hostage by a corrupt law firm
Drummond Woodsum is a scourge on this state and need to be fired from every school district they represent
Once again Maine “legislators” ignoring what 80% of us don’t want
Rather than treat mentally ill boys we subject our daughters to this gender bending nonsense
If you are female student athlete in maine you have no rights
Time for Maine voters/legislators to do the right thing and get this changed
School boards should request a second legal opinion
To say the federal government won’t be giving federal funds to individual schools
implying it is because that’s not how it works
Individual districts can apply for grants from the federal government so they are not being truthful
If they are a district who can demonstrate that they are in compliance with the federal title IX
Get a second legal opinion for crying out loud
MSMA and D W are not much more than a clanging symbol in an echo chamber
what if they want to organize their own private invitation only ‘girls only’ event
I’d be willing to bet there would be plenty of moral and financial support
Maine needs to address a huge mental health issue
Buckle up everyone gonna be interesting for the parents at these games
We need to direct our anger in force towards legislatures
Boys stay away from our girls or face domestic abuse charges for hitting a female
Maybe the girls need to join boys teams and hang out in their locker room
How pissed would parents and their girlfriends be
REFUSE TO PLAY…girls need to be a team and protest en masse
no facilities… it’s in the girls’ court
Fly those Maine Flags upside down and you girls need to boycott this bull$hit en masse
don’t accept your diploma when graduation comes around
Your refusal to engage in this madness is more powerful than all the money they make from it
They will graduate and move on never to have this experience again
We need to start the process to stop those in power creating this fiasco
Our babies deserve a positive life experience
Mills and Bellows disgust me.when do we stand up for our children… this is a conviction with meaning
After three generations of boys getting screwed
How about boys play against other boys and girls play other girls and trans play against other trans in sports
That’s neither what domestic abuse is
and the fact that someone would claim this shows how much of an anti-male crusade the VAWA actually is
They already are — it used to be called the Boy Scouts
I don’t see why boys can’t play field hockey
Maine did the right thing and stood up for fairness and inclusion
defied the Trump anti-trans bandwagon–and won
I hope this is an indication of a new trend
So Drummond Woodsum is in charge of Maine state athletics now
a far left progressive law firm advocating for mentally ill boys to compete against girls
So now we have a left wing “law firm” and a “non profit” (yeah right) running Maine sports
Perhaps the Maine Wire could check on the % of administrative fees the MPA is taking in for salaries
Or what the sponsors on thier website think about thier position on boys competing with girls….Hammond Lumber
I bet they’ll run for the hills if you ask
So let me understand this….if 2 high school girls are sitting on a park bench and some 18 year old guy comes and pulls his junk out in front of them he gets arrested
But that same guy can claim he’s a girl and the same 2 girls have to share a bathroom and Locker room with him where he is free to pull out his junk in front of them….yeah makes total sense
The “New Maine” where Women are Women and Men want to be Women too
Disgraceful State all because of a Lunatic in the Governor’s seat
We are funding this attack on our children
This and the teacher in Waterville are a showcase of what fools the body politic and public education system have made of the electorate
the fact it is still going on tells u we r getting played
cant justify why other than for thier own selfish reasons
its now up to the student athletes to stand for what is right
maybe they wont be getting played if they dont play the game
maybe if the top 1% didnt have all the money a local rich guy would put up 100 mill and make a non profit privite sports leauge…
The Maine Wire story on the Bonney Eagle alleged rape may be something to consider before turning their daughters over to this utter nonsense
“to participate on high school sports teams in accordance with their “gender identity.”
Gender delusion is the term for stating how you wish to be identified
as opposed to how you are identified in reality and scientific terms
Gender identity is not something you can make one choice on weekdays
wake up each day and choose what gender you will be for that day
It is like we in a Hunger Games state of mind
Me and Janet (see you in court) Mills love watching when a guy beats the living tar out of women in athletic endeavors
This crap would never have happened under a LePage administration
I have absolutely no respect for anyone who believes XY belongs in female sports
so now we have boys sports and misc sports
the women rights activists are no where to be found
they are the clowns allowing men to put set you back
I just started 3 weeks ago this web income system that my friend recommended to me and I’ve gotten 2 checks for a total of $9,200… this is the best decision I made in a long time
This extra cash n4550 has changed my life in so many ways
Here is I started_______ Tinyurl.com/homestar2?/n4550
For the record seventy two percent of high school graduates can not identify where girls pee from
So maybe it’s time to stop crying over genitalia
I mean clearly y’all missed the biology class that covers that but did you skip health class too
So here we are with a ton of 40+ men and women pouting because some sissy dude did indeed place better than a girl on a high school track team?!?
Every single kid in Maine who is homeschooled gets screwed out of the opportunity to even try playing sports
You know if you aren’t willing or can’t vaccinate 4-21 year old children in accordance with cdc recommendations then they are completely denied a right to public or private education/sports etc in Maine?!?
these fairy boys would have been delt with
is there any part of the RICO Statutes that apply to child abuse &/or endangerment in the public education system
For those with the bright idea of pulling another Charlie Howard, see: https://www.legislature.maine.gov/lawlibrary/what-is-maines-law-on-hate-crimes/9469
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Conduit Books will not ‘seek an adversarial stance … but the emphasis at first will be on ambitious
political and cerebral fiction by men that is being passed by’
A writer and critic has launched a new independent press that will focus on publishing books by male writers.
Conduit Books, founded by Jude Cook, will publish literary fiction and memoir, “focusing initially on male authors”.
Cook said the publishing landscape has changed “dramatically” over the past 15 years as a reaction to the “prevailing toxic male-dominated literary scene of the 80s, 90s and noughties”. Now, “excitement and energy around new and adventurous fiction is around female authors – and this is only right as a timely corrective”.
“This new breed of young female authors, spearheaded by Sally Rooney et al, ushered in a renaissance for literary fiction by women, giving rise to a situation where stories by new male authors are often overlooked, with a perception that the male voice is problematic,” he said.
These “overlooked narratives” might address fatherhood, masculinity, working class male experiences, sex, relationships, and “negotiating the 21st century as a man” – “precisely the narratives” that Conduit hopes to publish.
Cook said conversations about toxic masculinity after the second election of Donald Trump and the popular Netflix series Adolescence means that the “subject of what young men read has become critically important”.
whose book The Borrowed Hills was shortlisted for this year’s Sunday Times young writers’ award
what with the next generation of them simmering with rage online
but it’s a question I’ve been hearing for a long time
can sometimes be dismissed as unserious or unevolved
A book press willing to tackle that is a good start but the hard part will be cobbling together an audience out of readers who have become neglected.”
The press is actively looking for a book to launch with
“preferably a debut novel by a male UK novelist under 35”
with submissions open throughout the month of May
short story collections or memoirs a year beginning in spring 2026
It “can’t be over-stressed” that Conduit Books “doesn’t seek an adversarial stance”
“Nor is the press looking to exclude writers of colour
Men have not suddenly stopped reading and writing literary fiction, said Cook – rather, they are simply “not being commissioned”. He pointed to 2020 data suggesting that 78% of those in editorial roles in the publishing industry are women
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“Whenever I send out a novel to editors, the list [of names] is nearly all female,” a male agent told Johanna Thomas-Corr for an Observer piece in 2021
“But it’s not the gender makeup that bothers him
it’s the prevailing groupthink – the lack of interest in male novelists and the widespread idea that the male voice is problematic,” wrote Thomas-Corr
The declining prominence of young male writers has been a topic of debate in the publishing industry for several years, in part sparked online by a Times piece by James Marriott titled “Booker prize 2020 longlist: where are the new male hotshot novelists?”
There has “never been an independent publisher that champions literary fiction by men,” said Cook
“Which is not to say we won’t publish fiction by women in the future – but the emphasis at first will be on male authors
political and cerebral fiction by men that is being passed by.”
Cook is the author of the books Byron Easy and Jacob’s Advice
He regularly reviews books for outlets including the Guardian and TLS
and teaches creative writing at Westminster University
He said that since announcing the new press
the response “has been overwhelmingly positive
especially from female authors and women who work in publishing”
All 5 Releases
The number of cases involving male victims of stalking and harassment rose nearly 11 percent last year
while the number of incidents involving female perpetrators jumped 14.5 percent
data released yesterday by the Ministry of the Interior showed
Police agencies handled 2,806 stalking and harassment cases last year
The number of cases involving female victims fell approximately 3.7 percent to 2,427
while incidents involving male victims were up 10.8 percent to 379
Photo courtesy of the Ministry of the Interior
About 32 percent of the victims were aged 21 to 30
27.7 percent were aged 31 to 40 and 20.5 percent were aged 41 to 50
403 were female and in 123 cases the gender was not known
the number of cases involving male suspects fell 4.4 percent
while that of those involving female suspects rose 14.5 percent
About 25 percent of the suspected perpetrators were aged 31 to 40
20.5 percent were aged 21 to 30 and 19.9 percent were aged 41 to 50
64.1 percent were categorized as general stalking and harassment cases
while 35.9 percent were categorized as involving domestic violence
About 79 percent of the suspected perpetrators in the general cases were male
while the number of female suspects increased 8.5 percent
Eighty-five percent of the suspected perpetrators in domestic violence-related stalking and harassment were male
while the number of cases involving female suspects increased 26.5 percent
About 85 percent of the victims in domestic violence-triggered stalking and harassment cases were female
while cases involving male victims surged 39.6 percent
police agencies issued 1,867 written warnings for stalking and harassment
No warnings were issued in 939 cases because
the allegations did not constitute stalking and harassment or no perpetrators were located
Judicial Yuan data showed that courts issued restraining orders in 105 cases
representing 67.3 percent of all petitions filed by victims
Police agencies handled 143 cases in which a suspect violated a restraining order
Among the 27 petitions to remand suspects in custody
Sheridan County YMCA Executive Director Liz Cassiday told Sheridan Media
the Sheridan County YMCA was short on female camp counselors before the beginning of the camp season
Cassiday reached out and Sheridan County answered the call
the Y is searching for a few more male camp counselors to join the team.
Camp counselors are more than just chaperons watching over campers as they enjoy their summer season
mentors and make long lasting positive impacts on the lives of children.
The job of camp counselor also guarantees a summer job that is anything but routine
The position provides plenty of time outside and the chance to mentor children while they learn safe and time tested techniques on taking full advantage of living in the Bighorn Mountains
being a camp counselor offers numerous advantages
and a chance to positively impact children’s lives
It’s a rewarding experience that fosters leadership
all while enjoying the great outdoors and making lasting memories
To learn more and to contact the Sheridan County YMCA, click here.
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