Volume 10 - 2023 | https://doi.org/10.3389/fmolb.2023.1155579 The second messenger cyclic di-GMP (c-di-GMP) controls the transition between motility and sessility in many bacterial species by a variety of mechanisms including the production of multiple exopolysaccharides tomato (Pto) DC3000 is a plant pathogenic bacteria able to synthesize acetylated cellulose under high c-di-GMP levels thanks to the expression of the wssABCDEFGHI operon Increased cellulose production enhances air-liquid biofilm formation and generates a wrinkled colony phenotype on solid media We previously showed that under low levels of c-di-GMP the regulators FleQ and AmrZ bound to adjacent sequences at the wss promoter inhibiting its expression but only FleQ responded to the presence of c-di-GMP by activating cellulose production we advance in the knowledge of this complex regulation in Pto DC3000 by shedding light over the role of FleN in this process The distinctive features of this system are that FleN and FleQ are both required for repression and activation of the wss operon under low and high c-di-GMP levels We have also identified three putative FleQ binding sites at the wss promoter and show that FleQ/FleN-ATP binds at those sites under low c-di-GMP levels binding of c-di-GMP induces a conformational change in the FleQ/FleN-ATP complex allows promoter access to the RNA polymerase and leads to activation of wss transcription AmrZ is always bound at the wss promoter limiting its expression independently of FleQ FleQ is a c-di-GMP-responsive transcription factor that oppositely regulates the genes required for flagellar motility and surface adhesion in response to fluctuating intracellular levels of that second messenger c-di-GMP and ATP-bound FleN interact with FleQ to inhibit its ATPase activity and although the mechanism and the effect on transcription varies according to the promoter all three coordinate the regulation of flagellar motility and biofilm development promoting a sessile state of life to the detriment of a motile one our aim was to shed light over the possible interaction of AmrZ c-di-GMP and FleN at the Pto DC3000 wss promoter other compounds like antibiotics were added: gentamicin (2–10 μg/ml) rifampicin (10 μg/ml) and tetracycline (10 μg/ml) EPS production can be detected and even quantified using dyes as calcofluor (CF) or Congo Red (CR). CR binds to neutral or basic polysaccharides and some proteins, whereas CF is more specific and binds to β(1–4) and β(1–3) glycosidic bonds, like those present in cellulose, and positive colonies fluoresce under UV light (Spiers et al., 2002) Colony morphology and EPS production were visualized on MMR plates with Congo Red (50 μg/ml) and calcofluor (50 μg/ml) We generated the ΔfleN directed mutant by deleting most of its ORF (from nucleotide 10–816). First, a region with fleN adjacent sequences but lacking its ORF was amplified by PCR with specific oligonucleotides (Table 2) and cloned into pK18mobsacB (Schäfer et al., 1994) The plasmid was then electroporated into Pto DC3000 Transformants were selected in kanamycin (50 μg/ml) screened for sucrose sensitivity (15% [w/v]) and then grown in liquid LB at 4°C to force plasmid loss Cells were then plated on LB with sucrose (15% [w/v]) and the SucRKmS colonies which were expected to be double-recombinants were selected and check by PCR and sequencing Plasmid transformation of Pto DC3000 strains were carried out by electroporation. Electro-competent cells were prepared according to Choi et al. (2006) mixed with DNA (0.3–0.5 μg of DNA per ml of cell suspension) in 0.1 cm cuvettes and electroporated with a high-voltage pulse (1800 V) for 5 ms by using an Eppendorf electroporator 2510 Transformants were selected in LB agar plates supplemented with the appropriate antibiotics at least 3 biological replicates were assayed in octuplicate Pto DC3000 and mutants were grown on LB plates for 48 h and resuspended in sterile milliQ water 2 ml of cultures adjusted to an A660 of 0.05 in MMR were dispensed into wells of Nunclon Delta surface 24 microtiter polycarbonate plates (Nunc) and incubated at 20°C under aerobic and static conditions for 72 h The appearance of the bacterial communities at macroscopic level was studied after taking photographs directly from the plates Calcofluor binding assays by the different strains were performed as follows: bacteria were suspended from fresh LB plates in sterile milliQ water diluted into 10 ml flasks containing MMR supplemented with CF (50 μg/ml final concentration) to an initial A660 of 0.05 and incubated at 20°C under agitation for 24 h Cultures were then centrifuged for 15 min at 4000 rpm supernatant containing unbound CF broth was removed and the pellet was then suspended in 10 ml of distilled water CF binding measurements for 6 biological replicates of each strain were performed in a PTI fluorimeter (Photon Technology International) after confirming a similar growth of all strains and expressing the results in arbitrary units ± standard deviation AmrZ and FleQ were purified as described before (Prada-Ramírez et al., 2016; Pérez-Mendoza et al., 2019). For FleN purification, the One Shot BL21star (DE3) (pET28b(+):fleN) cells were grown at 28°C in 2-L Erlenmeyer flasks containing 1 L of 2 × YT culture medium (Sambrook et al., 1989) supplemented with kanamycin (50 μg/ml) Protein expression was induced at an A660 of 0.2–0.3 by adding 0.5 mM isopropyl β-D-1-thiogalactopyranoside and cultures were grown for another 5 h at 15°C when they were harvested by centrifugation at 5000 × g The pellet resulting from a 500 ml culture was resuspended in 25 ml of buffer A (25 mM Na-phosphate pH 7.0 5% glycerol) with protease inhibitor mixture (Complete™ Roche) and broken by treatment with 20 μg/ml of lysozyme and French press Following centrifugation at 13,000 × g for 60 min the FleN protein was predominantly present in the soluble fraction The supernatant was loaded onto a 5 ml Hi-Trap chelating column (GE Healthcare) equilibrated with buffer A and eluted with a gradient of a 50 mM-1 M imidazole Fractions containing His6-FleN were pooled and was dialyzed against 20 mM Tris-HCl pH 8.0 10% glycerol and stored at −80°C Protein concentrations were determined using the Bio-Rad Protein Assay kit A 458 bp fragment containing the wssA promoter region (wssA1-2) obtained from DC3000 chromosomal DNA by PCR was used as DNA probe (Table 2) The PCR product was isolated from an agarose gel by using the Nucleospin gel and PCR clean-up (Macherey-Nagel) and radiolabelled at its 5’-ends with [γ-32P]ATP and T4 polynucleotide kinase The labelled probe (20 nM) was then incubated with the indicated concentrations of purified FleQ FleN and/or AmrZ in 10 μL of STAD (25 mM Tris-acetate pH 8.0 3.5% (w/v) polyethylene glycol-8000 and 1 mM DTT) supplemented with 10 μg/ml of poly(dI-dC) and 200 μg/ml of bovine serum albumin The reaction mixtures were incubated for 30 min at 4°C and samples were run on 4% (w/v) native polyacrylamide gels (Bio-Rad Mini-Protean) for 2 h at 50 V at room temperature in Tris-glycine (25 mM Tris The results were analysed with Personal FX equipment and Quantity One software (Bio-Rad) Reactions (10 μL) were performed in STA buffer (25 mM Tris-acetate pH 8.0 1 mM DTT and 3.5% (w/v) PEG 8000) with 0.5 μM FleQ 4 Units of RNAse inhibitor (Roche) and 5 nM DNA template (wssA1-2 PCR fragment) After 30 min incubation at 4°C 0.5 U σ70-holoenzyme (New England Biolabs) were added and the reactions were incubated for 5 min at 30°C before the addition of 1.2 μL of the following elongation mixture: 0.1 mM each for ATP 0.05 mM UTP and 50 μCi [α-32P]UTP After a further 15 min incubation at 30°C the reactions were stopped by adding 3.7 μL of formamide sequencing dye Samples were electrophoresed in a 6.5% (w/v) polyacrylamide denaturing sequencing gel Statistical treatment of data was performed using R or Graphpad Prism 6 software Comparison among different strains or conditions was performed by one-way ANOVA with post hoc Tukey HSD test Biofilm development by Pto DC3000 mutant strains fleN and wssBC mutants were grown under aerobic and static conditions at 20°C in 24-multiwell plates for 72 h in MMR supplemented with tetracycline (10 μg/ml) when images were directly taken from the plate Serial dilution-based growth curves of the indicated strains where planktonic (left axes open symbols) or biofilm growth (right axes closed symbols) is plotted against the initial A600 of each dilution Circles represent the wild type strain and squares represent the fliC (a) Plots display one representative experiment of at least three biological replicates Error bars represent the standard deviation of the eight technical replicates the matrix of these biofilms is mainly composed of cellulose which give them a characteristic opaque and wrinkled appearance cellulose does not seem to be required for S-L biofilm development The fact that expressing fleN from a plasmid makes the colonies redder and slightly more fluorescent at low levels of c-di-GMP suggests a pleiotropic effect of FleN overproduction that is independent of FleQ but it may be related to flagellar assembly Colony morphology and cellulose production in the fleN mutant Representative colony morphology of the different strains grown in agar plates supplemented with Congo Red and Calcofluor in the presence and in the absence of pleD* Five μl of bacterial suspensions at A660 = 1.0 were placed on the surface of MMR plates with CR (50 μg/ml bottom) and pictured after incubation at 20°C for 3 days and then at 10°C for 5 days (B) Cellulose production at different c-di-GMP levels Pto and the fleN and fleQ mutants were grown in MMR with CF (100 μg/ml) for 24 h at 20°C and the fluorescence emission of the cell attached CF in liquid cultures was measured The graphs show the average amount of cellulose produced by the indicated strains in the absence (white bars) and in the presence of pleD* (grey bars) as fluorescence (in arbitrary units) referred to total cell protein Note that the wssBC mutant does not produce cellulose therefore the bars indicate non-specific CF retention Error bars correspond to the standard deviation of three biological replicates and a-c denote ANOVA categories with significant differences (p < 0.01) (C) Effect of c-di-GMP on the expression of the wss operon Total RNAs were obtained from bacteria grown in MMR at 20°C for 24 h Results show qRT-PCR of wssB in the wild type strain (Pto) and fleN and fleQ mutants with pJB3Tc19 (in the absence of pleD* white bars) or with pJB3pleD* (in the presence of pleD* Expression values were normalised with the housekeeping gene gyrA and referred to the wild type condition in the absence of pleD* The graph shows the average mRNA levels and error bars correspond to the standard deviation of four biological replicates; a-c denote ANOVA categories with significant differences (p < 0.01) these results suggest that FleN has a positive role in the regulation of cellulose synthesis in Pto DC3000 These results indicate that FleQ and FleN are both negative regulators of the wss operon and the positive effect of c-di-GMP on wssB mRNA levels is dependent on the presence of both FleQ and FleN FleQ seems to repress wss transcription under low c-di-GMP and activate it under high c-di-GMP levels and for both processes FleN is required indicating that FleN binds to FleQ to give higher-molecular-weight complexes and the ATP stabilizes the binding of the FleN-ATP/FleQ complex to DNA In vitro binding of the FleQ-FleN complex to the wss promoter region Binding reactions were carried out as described in Materials and Methods with 1 nM of labelled wssA1-2 fragment in the absence (-) and in the presence of 1 μM FleQ 0.5 mM of ATP and 0.5 mM c-di-GMP We showed before that c-di-GMP (but not c-di-AMP, ATP, or GTP) acts as an antagonist for FleQ repression at PwssA facilitating the release of FleQ from the promoter DNA (Pérez-Mendoza et al., 2019). Here we observed that c-di-GMP partially disrupted the FleQ-DNA complex (Figure 3 lane 3) but not the FleQ/FleN-ATP/DNA complex (Figure 3 lane 6) our EMSA results strongly suggest that FleQ binding to PwssA is promoted by FleN FleN alone did not detectably bind to the DNA (lane 7) Identification of the FleQ-FleN complex binding site at the wss promoter by footprinting analysis DNA probes corresponding to the wssA upstream region 5’ end-labeled on either the top or the bottom strand were prepared and incubated without (lanes -) and with FleQ (0.5 µm) and/or FleN (0.5 µm) ATP (0.25 mM) and c-di-GMP (0.25 mM) After partial digestion with DNase I or treatment with DMS and partial digestion with piperidine Nucleotide sequences protected by FleQ and FleQ-FleN in the absence of c-di-GMP (red) or by the FleQ-FleN complex in the presence of c-di-GMP (blue) are indicated Protected (arrow) and hyperreactive (*) nucleotides are also indicated (B) Localisation of the FleQ and FleQ-FleN binding sites at the wss promoter The boxes indicate the regions protected from DNAse I by FleQ or FleQ-FleN (red) and FleQ-FleN in the presence of c-di-GMP (blue) in the top and bottom strands The -10 and -35 regions and the wss transcriptional start site are in bold To localize the FleQ/FleN-ATP binding sites accurately and to establish the guanosine residues in close contact with the bound complex, methylation protection patterns were determined on both DNA strands using dimethyl sulfate (DMS) as a footprinting reagent (Figure 4A) Protection of guanosine residues by FleQ/FleN-ATP was observed at the region identified by DNase I footprinting thus confirming the location of the binding sites Protection from DMS methylation was detected at G-76 G-79 and G-90 at the top strand when FleQ was bound to the DNA either alone or in the presence of FleN (±ATP) G-76 and G-90 were protected in the absence of c-di-GMP (lanes 9 but less in its presence (lanes 10 and 13) hyperreactivity at Gs -88 and -89 was observed when FleQ was present in the reaction (lanes 22–26) but diminished in the presence of c-di-GMP (lanes 23 and 25) the binding of FleQ alone to the wss promoter is weakened in the presence of c-di-GMP the FleQ/FleN-ATP complex remains bound even in the presence of c-di-GMP as shown by the alterations of DMS protection and hyperreactivity patterns which indicate that the contacts have changed In vitro binding of the FleQ-FleN complex to the wss promoter region in the presence of AmrZ.Binding reactions were carried out in the absence (-) and in the presence (+) of FleQ Putative shifted protein-DNA complexes are indicated Location of all the regulators binding sites at the wss promoter DNA probes corresponding to the wssA upstream region 5’ end-labeled on either the top or the bottom strand were prepared and incubated without (lanes -) and with (+) FleQ and/or FleN Nucleotide sequences protected by FleQ and FleQ-FleN in the absence of c-di-GMP (red) by the FleQ-FleN complex in the presence of c-di-GMP (blue) or by AmrZ both in the absence and in the presence of c-di-GMP (green) are indicated The boxes indicate the regions protected from DNAse I by FleQ or FleQ-FleN in the absence of c-di-GMP (red) FleQ-FleN in the presence of c-di-GMP (blue) or AmrZ both in the absence and in the presence of c-di-GMP (green) in the top and bottom strands all the in vitro assays reveal that FleQ/FleN-ATP and AmrZ are able to bind to the DNA at the same time repressing the expression of the wss operon under low c-di-GMP levels but the remodelling of the FleQ/FleN-ATP complex on the DNA allows wss transcription Multiple round transcription assays were carried out as described in Materials and Methods The assays were performed in the absence (-) or in the presence (+) of FleQ c-di-GMP or c-di-AMP were also added to the reaction C- indicates a reaction without template DNA The 232 nucleotide mRNA synthesized from PwssA is point out by an arrowhead The graph shows the average amount of the mRNA produced as percentage of the condition without any protein (-) Error bars correspond to the standard deviation of six different transcription assays we can conclude that the FleQ and AmrZ binding sites are present in the Pseudomonas species bearing the wss cellulose synthesis operon suggesting that the regulatory mechanism may be also conserved in the syringae and fluorescens groups Since AmrZ and FleQ binding sites are adjacent they could interact when bound to the wss promoter and the conformational changes induced by FleQ/FleN-c-di-GMP in the DNA may disturb the binding and/or the activity of AmrZ as a repressor Since the role of FleN in Pto DC3000 cellulose production is so different from that in P future studies should focus on the molecular function of FleN in flagellar gene expression at Pto DC3000 This regulation should require the FleQ ATPase activity and σ54 a mechanism somehow opposite to that described for the regulation of the wss operon We anticipate that the analysis of FleN and c-di-GMP roles in the expression of flagellar genes in Pto DC3000 will most likely bring some surprises The original contributions presented in the study are included in the article/Supplementary Material further inquiries can be directed to the corresponding author M-TG designed research; FG and AL-S contributed to the conception and design of the work; LM-R All authors contributed to manuscript revision This research was supported by grants P20_00834 [funded by Plan Andaluz de Investigación Desarrollo e Innovación (PAIDI 2020) Consejería de Transformación Económica and by European Regional Development Fund (ERDF)] and BIO 2017-83533-P (funded by MCIN/AEI/10.13039/501100011033 and by “ERDF A way of making Europe”) Muñoz for their technical assistance The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fmolb.2023.1155579/full#supplementary-material Supplementary Figure S1 | Pellicle formation by mutant strains of Pto DC3000 under high c-di-GMP levels The shown strains with pJB3pleD* were grown under aerobic and static conditions at 20°C in 24 multiwell plates for 72 h in MMR supplemented with tetracycline (10 µg/ml) The pellicles formed were disintegrated in 2 ml of deionized water and 8 µl of this suspension was deposited on a clean glass slide and 8 µl of of a 1:1 mixture of 10% KOH (w/v) and calcofluor white stain (Fluka) were added to each sample A coverslip was deposited on the sample that was immediately examined in an epifluorescence microscope Leica DMI600B (Leica microsystems) using UV light excitation (365 nm) Supplementary Figure S2 | Colony morphology of the fleN mutant and its complemented strain Representative colony morphology of the indicated strains grown in agar plates supplemented with Congo Red and Calcofluor in the absence (- pJB3Tc19) and in the presence of pleD* (+ pBBRN) or with fleN expressed in trans from a plasmid (+ bottom) and pictured after incubation at 20°C for 3 days and then at 10°C for 5 days Supplementary Figure S3 | Multiple sequence alignment of regions upstream of the cellulose synthesis operon found in the Pseudomonas genus (A) Multiple sequence alignment of regions upstream of the cellulose synthesis operon found in the Pseudomonas genus The sequences shown are from Pseudomonas sp (B) Multiple sequence alignment of regions upstream of the wssA orthologs found in the P The putative FleQ binding sites are shown in blue (box 1) and red (boxes 2 and 3) and the putative 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Google Scholar Citation: Martínez-Rodríguez L FleN and c-di-GMP coordinately regulate cellulose production in Pseudomonas syringae pv Received: 31 January 2023; Accepted: 17 March 2023;Published: 27 March 2023 Copyright © 2023 Martínez-Rodríguez, López-Sánchez, García-Alcaide, Govantes and Gallegos. 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I was drawn to UNC Kenan-Flagler because of their healthcare concentration The breadth of course choices enable students to explore various dimensions of healthcare ranging from medical entrepreneurship to international immersion electives What has been your first impression of the Kenan-Flagler MBA students and alumni you’ve met so far Tell us your best Kenan-Flagler story so far UNC Kenan-Flagler embodies the Carolina Way that I have grown so fond of over the years I have come to expect and appreciate the collaborative learning environment that is cultivated amongst colleagues This year’s class of 2024 does not disappoint Although we have only had a few sessions in orientation I have been able to see the innovative thinking and leadership qualities in my colleagues My best UNC Kenan-Flagler story so far has been during Team Dynamics exercise with our legacy groups The 12 of us in a group were tasked with transporting ping pong balls into PVC tubes using only our wits and pencils The overall exercise was illuminating in that I was able to witness the resilience of our team when faced with obstacles and evolution of thinking and strategy as we ultimately mastered the task What course, club or activity excites you the most at Kenan-Flagler? I am most excited about the Student Teams Achieving Results (STAR) program in the spring The experiential course enables students to work with clients to craft solutions to a specific problem through the application principles of consulting frameworks and strategy Describe your biggest accomplishment in your career so far: As a medical student, I wear a “variety of hats” to enrich and add innovation to our curriculum through my roles in student government and beyond. My biggest accomplishment during my career in medicine is my work as a Paul A. Godley Art of Medicine Fellow Through a partnership with the Ackland Art Museum we curated a seminar series that analyzed race and medicine in the vehicle of art for all incoming first-year medical students This undertaking was particularly challenging because we were in the height of the Pandemic my co-fellow and I were able to transition to a virtual setting the series to a virtual landscape through our position as vice presidents of curriculum affairs we were able to work with administration to codify the programming into our medical curriculum or listened to that you would highly recommend to prospective MBAs There are various facets of business that are new to me so I set out to expose myself to media and literature that prepared me for the task at hand I would recommend the book Machiavelli for Women: Defend Your Worth Grow Your Ambition and Win the Workplace by Stacey Vanek Smith This work was provided a dynamic comparison of principles from “The Prince” and women entering the corporate workforce I have had a chance to apply a few strategies already that has been rewarding what would make your MBA experience successful so I gauge my success based on how well I accomplish them I know I will have hit my mark when I am able to fuse my interests in consulting strategy and clinically practicing medicine My ultimate driver in this process is understand the stakeholders logistics and strategy involved in holistically navigating the healthcare system What advice would you give to help potential applicants gain admission into Kenan-Flagler’s MBA program I would encourage potential applicants to reflect on their “why.” While I was applying for business school I would regularly practice introspection to figure out what intrinsically was motivating my pursuit of an MBA I was able to fashion a narrative of my passions involving affordable healthcare and equitable access DON’T MISS: MEET THE MBA CLASS OF 2024: INFLUENCERS & INNOVATORS Our Partner Sites: Poets&Quants for Execs | Poets&Quants for Undergrads | Tipping the Scales | We See Genius Website Design By: Yellowfarmstudios.com Flen graduated from basic military training at Joint Base San Antonio-Lackland eight-week program that included training in military discipline and studies Airmen who complete basic training earn four credits toward an associate in applied science degree through the Community College of the Air Force step-daughter of David Clay of Fayetteville She is a 2006 graduate of Seventy First High School She earned an bachelor's degree in 2010 from North Carolina Agircultural and Technical State University You may not be able to find the page you were after because of: You might find one of the following links useful: Discover the mipTOF for fast and high-quality trace element and metals analysis in the air The MPA Horizon Next-Gen Membrane Permeation Analyzer delivers advanced gas and vapor permeation analysis for membranes and barrier films With state-of-the-art Proton Exchange Membrane stacks PSM Series electrolysers ensure economical high-purity hydrogen production for large-scale applications ACCIONA's Turbine Made initiative recycles decommissioned wind turbine blades into high-performance surfboards Watercycle Technologies is transforming battery recycling and mineral recovery supporting the transition to a low-carbon economy you can trust me to find commercial scientific answers from AZoNetwork.com please log into your AZoProfile account first Registered members can chat with Azthena, request quotations, download pdf's, brochures and subscribe to our related newsletter content A few things you need to know before we start Read the full . Owned and operated by AZoNetwork, © 2000-2025 2022A still from “Descendant,” a documentary about a Black community that has long fought to preserve its history in the face of erasure.Photograph from Netflix Save this storySave this storySave this storySave this storyIn Margaret Brown’s documentary “Descendant,” a man named Anderson Flen walks through the streets of Montgomery and wonders aloud about the people who walked there before him people who had less freedom and fewer opportunities a freed Black settlement on the Gulf Coast founded by people who were brought over on the Clotilda the last known slave ship to reach the United States is working with community members and preservationists to transform Africatown into a tourist destination that honors the legacy of enslaved Black people The purpose of his trip to Montgomery is to visit the National Memorial for Peace and Justice widely known as the national lynching memorial which was opened in 2018 by the civil-rights lawyer and activist Bryan Stevenson and is an important reference point for Africatown’s efforts “The real test a lot of times is not in coming a few seconds.” He worries that the memorial will become “another form of entertainment.” whose center is three miles from Africatown She is the daughter of a reluctant débutante mother and a Jewish songwriter father she was frightened when her father pointed out houses owned by K.K.K warning her to be careful and know her place as a Jew But she had blue eyes and told me that she was “invisibly Jewish.” Her 2008 documentary “The Order of Myths,” examined the fraught racial and class dynamics of Mardi Gras in Mobile Her subsequent films focussed on the devastating effects of the 2010 Deepwater Horizon spill on working-class Gulf Coast communities as well as the voter-suppression efforts directed at rural African Americans in Alabama’s Black Belt she was exposed more deeply to the story of the Clotilda and the origins of Africatown—she didn’t remember learning it in school Brown set out to tell this story from the perspectives of descendants from both Black and white sides when she approached the family of the white man who’d paid for the voyage read by descendants in the places that Cudjo lived and worked objecting to her choice to phonetically render his Black vernacular speech The manuscript remained unpublished until 2018 the same year that Brown started shooting her film Meaher bought a schooner for a voyage to Ouidah where he funded the purchase of a hundred and ten young men and women they were forced off the ship and made to hide in the swamplands The schooner was burned and scuttled to destroy evidence of the voyage; Meaher would have faced the death penalty if discovered The captives were enslaved and put to work locally or sent to plantations farther away he and others sold many of them small parcels on which they founded Africatown an autonomous community where they grew food and taught the next generation their customs and languages The Meaher family is still one of the biggest landowners in Mobile and over the years they have leased their land to industrial plants that have polluted the land and contributed to a public-health crisis in the community according to research done by the Mobile Environmental Justice Action Coalition They have never apologized for the Clotilda voyage marine scientists confirmed the discovery of the Clotilda’s remains The discovery provided the Africatown community with scientific validation of their story (Researchers said last year that the ship was remarkably well preserved and may contain DNA traces.) Following the find Africatown held a bell-ringing ceremony attended by local politicians and members of national news outlets In a scene from “Descendant,” once the event has ended a former real-estate agent whose lineage stems from the Clotilda She fears that her community’s history will “be taken the same way our people were taken.” Of the nascent efforts to build a museum in Africatown and to redevelop the area Joycelyn Davis at the bell-ringing ceremony held in Africatown a few months after marine scientists confirmed the discovery of the Clotilda’s remains in 2019.Photograph courtesy the authorAt a community meeting in Africatown It showed what the Clotilda may have looked like including how the captives were packed into its hold The moment after the cover is whisked off and a small burst of clapping peters out the discomfort in the room is palpable as people take in the harrowing depiction of their ancestors lined up head-to-toe in coffinlike slats Many of the attendees’ faces appear filled with fear and regret “The pain and suffering that my people have had to endure throughout this whole process is a tremendous burden,” he says He proposes that “some dollars” come to those related to the Clotilda’s victims one of the divers who helped confirm the ship’s discovery affirms the community’s suffering and suggests “You all should come up with what justice means to you.” It is a complicated question with a variety of answers a descendant of the Clotilda survivor Charlie Lewis and dreams big about a similar monument in Africatown “I hope that it could be appreciated like this,” she says Emmett Lewis tells his young daughters stories about his great-great-great-grandfather Cudjo because he wants them to know what he stood for “My only fear is for my people’s story not to be told.” Tunstall worries about Africatown residents getting priced out as the region becomes a tourist attraction while expressing ambivalence about reparations I don’t think there’s anybody to punish,” she says a member of the Clotilda Descendants Association although news articles and books have come out about Africatown the community is still suffering economically and socially These attempts to grapple with what justice might mean resonate well beyond Africatown are watching to see how America’s history of racial violence will be memorialized the former director of programs at the History Museum of Mobile don’t know where their African ancestors came from “the African American community for one time can say a professor of history at the Harvard Kennedy School who studies truth and reconciliation efforts from Belfast to Rwanda believes that memorializing victims of structural racism is an important part of a larger movement of racial reckoning in the U.S but that memorials alone are “insufficient to the harder work of transforming a society.” These efforts don’t go far enough because they are too “passive” and easy to skip He cited the importance of Berlin’s Holocaust Memorial being placed in the heart of downtown and said that memorials need to “confront the spatial segregation that exists” and “penetrate areas that people cannot avoid.” A museum in Africatown would allow people to “opt out” of learning about the history of the Clotilda the civil-rights lawyer and founder of the national lynching memorial addressed this problem by adding a second set of steel rectangles to the memorial He invited the respective counties to claim their monuments and to establish a memorial on their home ground to lynching victims He also required each county to demonstrate that its community was taking steps toward economic and racial justice before acquiring its column The unclaimed monuments that remain on display at the national lynching memorial serve as a reminder of the lack of redress across the country “I still don’t know what my idea of justice is.” Lonnie Bunch the secretary of the Smithsonian Institution and founding director of the N.M.A.A.H.C. reparations would mean providing “educational opportunities for all,” as such opportunities would be what “would make so many of those enslaved ancestors smile.” Muhammad argued that school is the best channel for “acceptance and acknowledgment” to be disseminated in society Yet the increased efforts to present schoolchildren with the truth about America’s racial history have resulted in laws limiting what can be taught passing or getting introduced in at least forty state legislatures “Putting up a monument might actually be easier anywhere in this country than changing how we teach each other and our children about what happened,” Muhammad said a preservationist and the director of the African American Cultural Heritage Action Fund at the National Trust told me that historic preservation is “a form of repair and a way to correct past injustice.” For Leggs a historic site has the unique ability to reduce the gap between space and time Brown said that the aim of her film was similar: to illustrate that “the past is in the present cemented a narrative that positioned whites as heroes and erased the contributions of Black people Leggs hopes that a “greater reverence for Black history” could lead to a “greater reverence for Black bodies and Black people.” Mary Elliott the curator of American slavery at the N.M.A.A.H.C. and class divides express frustration at not having been taught this history “Descendant” is not only about justice but also about memory As Americans struggle to come to terms with the atrocities of racial violence these questions are contentious and uncomfortable The only certain thing is that we cannot look away A long-ago crime, suddenly remembered A limousine driver watches her passengers transform The day Muhammad Ali punched me What is it like to be keenly intelligent but deeply alienated from simple emotions? Temple Grandin knows The harsh realm of “gentle parenting.” Retirement the Margaritaville way Fiction by F. Scott Fitzgerald: “Thank You for the Light.” Sign up for our daily newsletter to receive the best stories from The New Yorker. The King and Queen attended the World Music concert at Stenhammar Palace included musical performances by Flen World Orchestra Music can bring together people from difference cultures and generations allowing them to understand each other despite their different languages and frames of reference The concert included musical performances by Flen World Orchestra World Music at Stenhammar was first held in September 2019 The concert was arranged by Stenhammar Estate Find out more about how we use cookies Please enable JS and disable any ad blocker The project is expected to be completed by spring of 2025. 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Please upgrade your browser or activate Google Chrome Frame to improve your experience Swedish thin film solar manufacturer Midsummer has selected a factory in southeastern Sweden to develop its new solar module production facility which will produce copper indium gallium selenide (CIGS) solar cells for the European market Swedish manufacturer Midsummer has selected a location for its forthcoming 200 MW factory that will produce CIGS thin film solar cells for the European market The factory, which is being partially funded by a €32.3 million grant from the EU Innovation Fund, will be located in the municipality of Flen, southeastern Sweden Midsummer will take over the premises this coming September provided the EU approves the location and start of the project says Flen fulfills all of the company’s requirements reliable electricity supply and established local infrastructure that is favorable to this type of production “We feel that we have received and will receive strong support from the Flen municipality for our investment Flen is also geographically well located for the transport of input materials and finished products across Sweden and all of Europe,” Jaremalm added the factory will employ around 200 people once fully operational Midsummer’s product range focuses on light and environmentally sustainable solar panels for primarily low-load-bearing commercial and industrial roofs that cannot withstand the weight of traditional silicon panels Last month, the company announced a partnership with France’s Soprasolar which will see its thin-film panels integrated into Soprasolar’s roof membranes to be made available to customers in France and other markets More articles from Patrick Jowett Please be mindful of our community standards and website in this browser for the next time I comment Δdocument.getElementById( "ak_js_1" ).setAttribute( "value" By submitting this form you agree to pv magazine using your data for the purposes of publishing your comment Your personal data will only be disclosed or otherwise transmitted to third parties for the purposes of spam filtering or if this is necessary for technical maintenance of the website Any other transfer to third parties will not take place unless this is justified on the basis of applicable data protection regulations or if pv magazine is legally obliged to do so You may revoke this consent at any time with effect for the future in which case your personal data will be deleted immediately your data will be deleted if pv magazine has processed your request or the purpose of data storage is fulfilled Further information on data privacy can be found in our Data Protection Policy Δdocument.getElementById( "ak_js_2" ).setAttribute( "value" This website uses cookies to anonymously count visitor numbers. View our privacy policy. × The cookie settings on this website are set to "allow cookies" to give you the best browsing experience possible If you continue to use this website without changing your cookie settings or you click "Accept" below then you are consenting to this Close .st1{fill-rule:evenodd;clip-rule:evenodd;fill:#2a2a2a}By Graham Couch | gcouch@mlive.comErik Holladay / Special to the Kalamazoo GazetteFlenard Whitfield known as "Flen" to his teammates and coaches is considered WMU's go-to guy as its first season post-David Kool begins.KALAMAZOO — Flenard Whitfield knows what’s coming He saw it happen to David Kool almost every night the past two seasons and watched as Kool struggled against it and then flourished in spite of it Whitfield is about to experience first hand the less pleasant side of respect On a Western Michigan University men’s basketball team lacking proven scorers Whitfield is likely to be the beginning of every defensive scouting report “You have to put yourself in the other team’s shoes,” Whitfield said “You have to watch film on yourself sometimes to see what would you do against yourself if you were the other team ‘How would I get out of that double team?’” “I don’t think I want to tell anybody how I would stop myself,” Whitfield said There is little doubt as WMU embarks on the post-Kool era that much of its success is tied to Whitfield His 10.1 points and 5.2 rebounds per game last season are about double the averages of any other returning player Whitfield became a bona fide scoring threat in the post late last year putting up 12.2 points and 6.3 rebounds over the final nine games of 2009-10 “I’ve just seen the intensity out of him,” sophomore forward Nate Hutcheson said he’ll tell you and he’ll tell you in the right way I think everybody just kind of looks to him as a leader Leadership is not coach Steve Hawkins’ concern It’s the group he’s asking Whitfield to lead “This team is very immature,” Hawkins said In the seven years I’ve been here as the head coach and they run around pretty well and they play hard but things can turn on a dime in the middle of a practice They can turn a practice into a complaining session about each other in a hurry trying to keep them on task and trying to get to the next thing the speed of the leader is the speed of the gang.” Whitfield’s own “emotional maturity,” as Hawkins put it could determine what sort season he and the Broncos have The Detroit native spent much of his first two seasons plagued by foul trouble will his frustration result in what Hawkins deems “immature fouls.” “He can’t pick up the cheap fouls,” Hawkins said “He’s had a tendency to get into a little battle within the battle and he’ll chuck somebody as he’s coming down the court or pop somebody or stick an elbow out as somebody’s coming off of a screen to catch them Those are fouls he could commit in the past and I’d bring him over the bench and I’d be like And that leads to Hawkins’ other fear: “It’s easier for a defense to take away a big guy.” we can put the ball in your hands in a variety of ways All you have to do (against a big guy) is put a guy in front of him and a guy behind him and he’s gone “That opens up other things where other players have to step up That’s why I think it’s so important that the other players establish themselves.” Whitfield insists this crew is more equipped to help him than he and his teammates were two years ago to help Kool We’ve got big Stain (freshman center Matt Stainbrook) athletic body that weighs probably 270 still so it’s hard to focus on just one guy this year.” Other teams will probably wait to see it to believe it “It definitely feels like my group,” Whitfield said you’ve got to pick them up.’ If anything goes wrong Contact Graham Couch at gcouch@kalamazoogazette.com or 269-388-7773 Follow him at twitter.com/broncosinsider or facebook.com/broncosinsider Use of and/or registration on any portion of this site constitutes acceptance of our User Agreement, (updated 8/1/2024) and acknowledgement of our Privacy Policy, and Your Privacy Choices and Rights (updated 1/1/2025) © 2025 Advance Local Media LLC. All rights reserved (About Us) The material on this site may not be reproduced except with the prior written permission of Advance Local Community Rules apply to all content you upload or otherwise submit to this site YouTube's privacy policy is available here and YouTube's terms of service is available here Ad Choices A mosque in Sweden was vandalized Sunday when unknown attackers painted a swastika on one of its door The Al-Huda Islamic Cultural Center’s mosque that came under attack is in Flen town of Sodermanland district Sweden Islamic Federation President Tahir Akan told Anadolu Agency the Muslim community was sad after the attack on their mosque ''Police have launched a large-scale search I hope those who did this would be caught soon The Flen mosque -- bought by the Al-Huda Islamic Cultural Center -- used to be a church; it was converted into a mosque in 2012 Flen Mosque’s keeper Abdi Osman Abukar told Swedish media the mosque had been attacked four times between 2012 and 2014 Developer SENS has secured a 30-year land lease for a 40MW battery energy storage project in Södermanland The deal has been struck with a landowner outside Bettna in Flen municipality Sustainable Energy Solutions Sweden Holding AB (SENS) said SENS will now aim to continue the development process of the project up to ready-to-build (RTB) status at which point it will sell control of the project to another developer or long-term owner That will follow the model it opted for in May this year when it sold a co-located 25MW battery storage project in Värmland country to Swiss renewable energy firm Axpo set to start construction in the second half of 2023 SENS didn’t give a timeline on when it expected the Södermanland project to reach RTB or be sold but it may be worth noting that it announced the land lease for the Värmland project in January five months before it sold the project to Axpo commented: “It is with pleasure that we have now signed another land lease agreement which represents a crucial first step as we now aim to establish a new battery storage facility Interest in battery storage has been impressive both as an integral part of renewable energy production and as a resource for frequency regulation.” The company has been active this year in starting new projects across Sweden it announced it was starting the development of a new 30MW battery storage project in Filipstad the same municipality where Axpo’s located The same month saw it announce a feasibility study with brewer Spendrup for a solar and storage project at a brewery in Grängesberg Opportunities for battery storage in Sweden have increased in the last few years as the country’s hydroelectric fleet’s ability to provide ancillary services begins to be exhausted and energy trading opportunities increase as more renewables come online and market volatility grows Subscribe | Login Ideas are popping and confidence is swelling You can’t wait to dive into business school Flen us a “Triple Tar Heel.” Already holding a BA (Biology) and MA (Physiology) from the university Flen witnessed the “innovative thinking” and “leadership qualities” that have been the hallmark of her UNC experience “The best UNC Kenan-Flagler story so far has been during Team Dynamics exercise with our legacy groups The overall exercise was illuminating in that I was able to witness the resilience of our team when faced with obstacles and evolution of thinking and strategy as we ultimately mastered the task.” Aidan McConnell jokes that his introduction to Kenan-Flagler involved “hanging by a thread – quite literally.” At a retreat for Vetter Dean’s Fellows he participated in the “Flying Squirrel” – a team-building exercise on a high-ropes course classmates work together to propel each other 50 feet in the air holding ropes so peers can experience what it is like to soar like Superman McConnell’s classmates were equally supportive when his feet were planted on the ground peers I’d only known for a few hours were already coordinating study groups offering their cars to help others carpool to the McColl Building and helping newly arrived international students acclimate to life in Chapel Hill I’m proud to be a part of this encouraging but in Chapel Hill they have a term for it: “The Carolina Way.” In sports the Carolina Way means being supportive and inclusive community ever committed to individual and team excellence “The Carolina Way” is more than a fluffy slogan It is the expectation that students carry for themselves – and their peers “They really embrace the spirit of the Carolina Way,” writes Mary Paul Msemwa “Everyone I have met has shown genuine interest in me and offered to assist me whether by giving their time out of their busy schedule or connecting me to someone who can assist or a resource I can use UNC MBA students are highly motivated high achievers but also know how to have a great time.” UNC’s mascot Rameses strikes a pose out of the Business School the Kenan-Flagler Business School also boasts two of the top MBA programs in Healthcare and Real Estate It is back by one of the country’s research universities while also maintaining two of the highest-ranked undergraduate business and online MBA programs The program is experiencing an impressive growth spurt enrollment has doubled over the past decade the school will be cutting the ribbon on a 140,000 square foot building that will enable it to double its undergraduate populate That doesn’t count Kenan-Flagler expanding its Executive MBA program to Charlotte’s Legacy Tower The Class of 2024 is packed with high potentials and seasoned practitioners alike He led a gas and oil drilling project that produced one of Pakistan’s largest gas reserves starting with feasibility and planning before moving onto drilling “I headed a 14-member team to safely drill part of the gas well that led to the gas discovery in Northern Pakistan,” Rahman notes The discovery was celebrated throughout the country since Pakistan is facing a serious energy crisis and thus was the perfect farewell I could have asked for before starting my MBA.” Numbers also reflect a storyline about the Class of 2024: They are influential and impactful Mary Paul Msemwa served as a liaison between the American and Tanzanian government resulting in $8.7 million dollars in tax refunds being repaid Raquel Argenal Matheu managed a $143 million dollar book of insurance loans – one of the largest for its West Coast operations He worked on a $280 million-dollar corporate bond issuance – the largest in the history of Nigeria a nation with the largest GDP and population on the African continent Lindsay Buchler spearheaded the re-organization of a shared services center that resulted in $3 million dollar savings in labor costs “We provided client executives with recommendations through quantitative analysis that aligned the desired shared services future state with the overall corporate strategy which ultimately resulted in a successful launch of the shared services center The success of this project and others was recognized by leadership which allowed me to achieve my promotion to senior consultant and the trust to lead several other impactful projects.” Hemadri Singh Kuntal also made her name at Deloitte as a healthcare IT consultant her sister battled COVID-19 as a physician-in-charge at a frontline hospital And Singh Kuntal soon joined the fight on a different front “I led a team of three executive clients and two consultants to proactively implement logic-based flags for potential COVID-19 patients in the electronic health record (EHR) This feature ensured that all healthcare workers operating in 800+ hospitals/clinics across the United States were alerted of potential COVID-19 patient around or near them.” Alexis Flen notes that she wears a “variety of hats” at the UNC School of Medicine – including class president was promoted to being an assistant vice president at Barclays where she served as a strategic finance partner in the London office Irasema González-Gutiérrez head up its “Women in Operations” efforts “It focused on mentoring women across Amazon’s North America South Central (NASC) region and launching inclusion events such as the first NASC “Women @ Amazon” virtual conference during the pandemic Amazon recognized the importance of the “Women in Operations” initiative due to the exponential improvement of women in the warehouse transforming our team to be 65% female-led in the sort-center network.” The Class is equally eclectic outside the office Aidan McConnell worked as a ballroom dancing instructor Hemadri Singh Kuntal’s passion is jump rope: In 2 minutes she can switch between eight styles of jumping rope who holds master’s degrees in special education and educational leadership Abdur Rahman can speak four languages – which he uses with different audiences The 2021-2022 application cycle represented a high-water mark for the Kenan-Flagler MBA. During the cycle, the school received 1,975 applications, a 32% jump over the previous year (though more in line with the 1,903 applications it received for a spot in the Class of 2022) This year’s class also represents a return to normal after COVID wreaked havoc on admissions After class size peaked at 344 students two years ago it has shrunk to 243 students in the Class of 2024 This is more in line with the 250 MBAs who traditionally populate the full-time class The smaller class size has also yielded another change: Kenan-Flagler has grown more selective After accepting nearly 53% of applicants in 2019 you’ll find MBAs who bring higher GMAT scores to Chapel Hill The Class of 2024 averaged a 706 GMAT – a full 10 points higher than the previous year (with scores ranging from 680-730 in the 85% range) as scores ran from 312-327 in the same range the percentage of women continues to climb rising from 34% to 38% with the current class International students account for 35% of the class a nine-point improvement from the previous year Another difference between the Class of 2024 and its predecessor The difference was made up by Business and Economics majors whose representation plunged from 49% to 31% Arts and Humanities majors compose the remaining 25% of the class the largest percentage of the class – 19% – last worked in Healthcare Financial Services professionals hold 17% of class seats which hold 5% or lower shares of the class Chances are, the Class of 2024 won’t be staying in these fields, observes Olivia Koziol, a 2022 grad and P&Q Best & Brightest MBA “Our MBA program is full of career switchers of every industry The reason UNC Kenan-Flagler is so collaborative is because many of us have a background different than the subjects we are learning about so we aren’t afraid to ask questions and push our boundaries this helps us grow from each other’s experiences.” Next Page: Interview with Kenan-Flagler Leadership Page 3: Profiles of 12 Members of the Class of 2024 More than half of Sweden’s land area is covered by forests and the forest industry here is a cornerstone of the country’s economy pulpwood and paper for hundreds of millions of krona each year and the industry accounts for roughly 10 percent of Sweden’s total exports The forest industry here grew rapidly the past century but as companies started cutting down more and more trees towards the middle of the 20th century some foresters began worrying that we were cutting down trees faster than they were growing and that that would lead to a nationwide shortage of wood they started searching for a faster growing tree and eventually found the North American Lodgepole pine a forest ecology consultant and associate professor of forest ecology at the Swedish University of Agricultural Sciences “They basically said that ‘we think we may need more wood in 35 years or so and this is growing faster’ and decided to grow it here,” says Engelmark was introduced in Sweden on a large scale in the 1970’s and today it covers more than 600,000 hectares that’s an area about the size of Sweden’s largest lake The Lodgepole pine has about 30 percent more needles than the native Swedish pine trees which could affect the ecosystem says Ola Engelmark who showed me around a small private plantation a few hundred metres from a busy road close to the city of Flen southwest of Stockholm “If this is allowed to grow and if we import more exotic species to Sweden then we will change the native ecosystems to a foreign ecosystem and then our native species won’t do well in these changed ecosystems The Lodgepole pines change the living environment for insects On top of that we also have to follow the Rio Convention which says that we have to strive to conserve our biodiversity we can’t just introduce a species only because it grows quicker,” says Engelmark species have a tendency to spread quickly and invade native ecosystems the North American pine tree could only be found at large-scale plantations in the northern parts of Sweden – as they were never introduced to the southern part of the country because the native spruce trees were already producing enough wood – but today the species is spreading in a country where the pine tree is the second most common tree the native Scots pine is in no way threatened by its North American relative any time soon still Ola Engelmark believes that the Lodgepole pine could become an increasingly common sight in Swedish forests in a near future “In the 1970’s many people said that this species wouldn’t regenerate here in Sweden It has the potential to grow almost everywhere but it has the ecological capacity to do that We can also see that it is invading certain parts of Sweden and I think we can foresee a kind of ‘contortafication’ as the Lodgepole pines here grow older and start producing more cones This will cause even more natural regeneration,” says Engelmark Canada: Canadian province of Quebec announces plan for northern development Finland: Finland forest sell-off has risks Russia: Counting elusive Finnish forest reindeer in Russian Karelia Sweden: Sweden’s Social Democrats make forest investment pledge For more news from Sweden visit Radio Sweden .st1{fill-rule:evenodd;clip-rule:evenodd;fill:#2a2a2a}By Graham Couch | gcouch@mlive.comMelanie Maxwell / AnnArbor.comFlenard Whitfield here against Eastern Michigan's Brandon Bowdry last Sunday won't play in today's game at CMU.KALAMAZOO — Contact Graham Couch at gcouch@kalamazoogazette.com or 269-388-7773. Follow him at twitter.com/broncosinsider Azuki has announced that it is adding nine new manga series, which include Star Fruit Books’ The Horrors of Noroi Michiru that will be a digital exclusive on the service Other newly added titles include Different Nation Some titles have been added from the publisher MediBang All nine titles are now available worldwide (except Japan) on the Azuki website and iOS and Android apps The Horrors of Noroi Michiru is an Azuki digital exclusive and will only be available to read on the service Star Fruit Books will release the series in print in two volumes Volume 1 is 204 pages and goes on sale July 18 Here are the details for Azuki’s latest manga licenses: the master of horror Noroi Michiru is available in English This two-volume collection includes his one-shots with each page rendered in exquisite detail to showcase his splendid artwork — each panel worthy of being framed Noroi Michiru’s aesthetic is impressive in that it conforms to Western expectations both amongst illustrated covers and also pages of narrative — focused artwork and narratives are as admirable as the efforts of Junji Ito Tsurukawa also has two one-shots available from Star Fruit Books on Azuki: Sawanabe Zombie and Ikyoudo Since the aliens invaded Earth 20 years ago they have co-existed with human beings in this country an institution with a mission to prevent violent crime happening in the invaded nation is the main protagonist of this full-colored sci-fi human drama confronts a crime organization in the bizarre but thrilling “Different Nation.” “Things I wanted to re-do seem to be endless…” An elite Office Lady has a unique ability to do minimum time looping but the limit suddenly turned into 10 years when she got in an accident The one who greets her in the high-school-past was that man who had lived in Komori’s heart for a long time… sometimes remembered as a smiling nerd sometimes as a lifeless body that Komori couldn’t save All is well if they’re not criminally inclined so a group of sinners is now fighting back for their right to try and not just be labeled at birth Chika grows medicinal herbs and sometimes helps treat her fellow villagers she notices someone dressed from head to toe in black standing in her herb garden and she also seems to be extremely knowledgeable about medicines and herbs Chika invites her in for a drink and asks her where she’s from Flen replies with the mysterious answer “over the eastern mountain it seems there is a secret behind Flen’s amazing medicinal knowledge In Izumi Nakama’s grade are twins Satsuki and Ibuki Asakawa unlike his stoic and cool younger twin Ibuki Izumi attracts the brother he was not interested in getting to know Izumi starts realizing Satsuki’s true character… (Note: Spiciness 1 out of 5) (Warning: uses of anti- gay slurs) “Rule 1 Series”: Rio and Etsu have been together since high school and now live together as a happy couple Madly in love and drunk in their attraction to each other their intimate lives together are a bond nobody can break After hearing about his father’s hospitalization Shouta decided to leave his Pastaio job in Italy and inherit his family Western cuisine restaurant in Hurubita Shopping District except probably his-used-to-be-innocent childhood friend who had now bloomed into an attractive guy everyone in the district wanted to sleep with Makoto’s casual sexual relationship with many young men in the district kinda drives Shouta crazy but maybe… Just maybe Makoto still saves some of his innocence for the guy who hadn’t been home for 7 years Keep your guard up and hunt everything you can because you never know when you’ll be hunted The high school teacher Kanichi Makabe is what you might call a “perfectionist” He does not compromise anything that might hinder his perfect routine From fighting over seats in the train to protecting a corner of an eraser every peaceful boring day is indeed a battlefield for him… the evolution that he and his team witnessed was repeatable The researchers didn't set out with the goal to evolve hyperswarmers but they did passage Pseudomonas aeruginosa on special plates over a period of days bacteria that could spread out had an advantage in harvesting nutrients from the surface some of those bacteria started hyperswarming Investigation of the bacteria showed that P aeruginosa gained its hyperswarming ability through a single point mutation in a flagellar synthesis regulator (FleN) were locked into a multi-flagellated state They became better at moving around to cover a surface the researchers saw this new ability independently arise 20 times Part 1: Swarming by the ancestral (wild type) strain shows the typical branching pattern of swarming colonies shows a very distinct phenotype without branching Part 3: Repulsion assay for hyperswarmer clone #5 shows that the colony is still repelled by the presence of a immotile strain (flgK-) although less than the wild type or other hyperswarmer clones "The fact that the molecular adaptations were the same in independent lineages suggests evolution may be The findings may be very important because biofilms are a major problem in clinical settings Infectious biofilms are hard to remove and difficult to kill with antibiotics Drugs that target FleN or that otherwise make bacteria better at spreading out and worse at settling down could leave them more vulnerable to antibiotics and easier to get rid of pathogenic bacteria might sound like the plot of a horror film but such bugs really have repeatedly evolved in a lab and the good news is that they should be less of a problem to us than their less mobile kin are also much worse at sticking together on surfaces in hard-to-treat biofilms They might even help us figure out a way to develop anti-biofilm therapies for use in people with cystic fibrosis or other conditions say researchers who report their findings in Cell Reports Source: Cell Press It’s been revealed that in order to meet the demand for goat cheese, or chèvre, Swedish farmers are killing off thousands of baby goat kids every year. To make goat cheese, you need goat milk, and for that you need mother goats. But all those little goat kids want to drink the milk, which farmers here seem to feel defeats the whole purpose of the exercise. So, according to figures from the Swedish Board of Agriculture, every year at least 6000 goat kids are killed here, so there will be more milk to make cheese. Most of them are males, since they will never produce any milk, and most are killed when they are just a few hours or maybe a day old. Their bodies are then burned or used for compost. But not every goat farmer here is comfortable with the system. Carmen Wikström raises goats at a farm outside Flen. “It feels like such a waste,” she tells Swedish Radio News. “To take a life just when it’s gotten started.” The dairy goat industry is booming in Sweden. Today there are around 60 goat farms here, compared to 20 in the mid-90’s. In the last eight years alone the number of goats here has doubled. Kerstin Jürss is president of Sveriges Gårdsmejerister, the Swedish Dairy Farmers’ Association, not to be confused with the much larger Swedish Dairy Association. Her organization brings together around 20 dairy farms in southern Sweden, and she says it is the increase in demand for goat cheese that is behind the problem of killing off the goat kids. “This ethical issue is a big thing because many think that it is tough to just breed, kill and bury,” she says. “Partly emotionally, but also ecologically. You don’t do this with any other animals really, because there you have a market for the meat.” Goat farmer Carmen Wikström has succeeded in finding a restaurant that buys her male goat kids, so she can pasture them for a few months before they are slaughtered. But the more her business grows, the more buyers she’ll need to find. Sometimes, she admits, she thinks about quitting goat farming. “I think about it,” she says. “I ask myself several times a year. Do I really want this?” Daily news podcast Mon - Fri at 4.30pm, plus weekly summary on Fridays at 4.30pm on P2 (P6 89.6FM in Stockholm), repeated on Mondays at 4.30pm on P6. Kontakta gärna Sveriges Radios forum för teknisk support där vi besvarar dina frågor vardagar kl. 9-17. I'm tryna hit him in his noggin' (Frrt I be tryna slide like a toboggan (Ha)Rich as fuck still eatin' Top Ramen (Ha)[Chorus]The bag in I'm in I know niggas cannot fuck with me (The bag in I'm in)The bag I'm in I make these niggas uncomfortable (The bag in I'm in)The bag I'm in I might put a nigga in a trunk (The bag in I'm in)The bag I'm in I make fifty thousand every month (The bag in I'm in)The bag I'm in win after win (The bag in I'm in)Can't be everywhere at once I wish I had a twin (The bag in I'm in)The bag I'm in ten after ten (The bag in I'm in)'Cause we keep them chickens in like Big Meech and Tee Flen' (Ha the bag in I'm in)This bag I'm in this one really personal (This bag in I'm in)This bag I'm in with this bitch I'm surgical (This bag in I'm in)This bag I'm in I don't need no friends (This bag in I'm in)Thank God for my ends To learn more, check out our transcription guide or visit our transcribers forum Find answers to frequently asked questions about the song and explore its deeper meaning