implementation and coordination of the department’s fundraising programs and major gifts in consultation with the Director of Athletics as well as the UNI Foundation and university advancement He is also in charge of the preparation and presentation of major gift proposals to current and prospective donors and maintain contacts with individuals and businesses nationwide for increases in current support or new major gift support to address athletic program needs Iowa native and 2004 graduate of nearby Wartburg College has spent nearly two decades in various roles in collegiate athletic leadership and administration He joins the Panthers after a ten-year run at the University of Arizona in Tucson where he has served as Senior Associate Athletic Director for Student-Athlete Experience and Excellence Mosbach was responsible for assessing the overall academic and student-athlete development programs’ effectiveness and provided guidance to matters of budgeting human resources and strategic planning for Wildcat students A member of the Intercollegiate Athletics (ICA) Leadership Team he was a member of Arizona’s NIL working group that created “Arizona Edge,” which served as an educational and information platform for NIL He also managed the department’s “5980 Academic Fund,” which provided student-athletes additional education related benefits totaling $3 million Mosbach also assisted the University of Arizona’s development office and the U of A Foundation in securing major gifts including a $5.95 million gift for the athletic department that included collaborations with two other units on campus as part of a large comprehensive $14 million gift to the university He also served as the secondary sport admin for Wildcat baseball and soccer having previously overseen directly the women’s tennis and both men’s and women’s swimming and diving programs Mosbach spent six years as Arizona’s Associate Athletic Director for Academics within the athletic department’s comprehensive student service program Commitment to an Athlete’s Total Success (C.A.T.S.) Mosbach managed a team of 15 staff members including academic counselors learning specialists and administrative associates and technology coordinators in helping improve Arizona student-athletes' academic success by way of improved cumulative grade-point averages (GPA) academic progress rates (APR) federal graduation rate and other NCAA and institutional data and standards of evaluation Mosbach spent eight years at the University of Southern California in Los Angeles in its academic services office before rising to assistant director within two years and becoming Assistant Athletic Director for Student-Athlete Academic Services in 2011 he helped lead the academic advising team for Trojan student-athletes in their academic endeavors coordinated summer orientation and summer bridge programs for athletes and helped consulate with the university’s Director of Student Services to help guide students through financial aid After graduating from Wartburg in 2004 as a member of the Knights’ football team with a Bachelor of Arts degree in Business Administration with an emphasis in sport management Mosbach spent two years as a graduate assistant in academic advising at the University of Tennessee in Knoxville graduating with his Master of Science degree in Sport Management in 2006 John Mosbach named Senior Associate Athletics Director1/24/2025 3:37:00 PM | General Longtime sports administrator joins senior athletics department staff CEDAR FALLS, Iowa --- The UNI Athletics Department on Friday announced the hiring of John Mosbach as Senior Associate Athletic Director for Development and NIL implementation and coordination of the department's fundraising programs and major gifts in consultation with the Director of Athletics He will also be in charge of the preparation and presentation of major gift proposals to current and prospective donors "I am thrilled to announce the hiring of John Mosbach to the Panther Athletics family as he rounds out the Athletics Senior Staff team," said UNI Director of Athletics Megan Franklin "His executive experience will lend well to the betterment of the department John is a native Iowan who will immediately engage and connect with friends of the department John's experience means he is a connector which will lend well to supporting the donor experience as they entrust the department with legacy giving decisions." "I am honored to join the University of Northern Iowa and contribute to the continued success of Panther Athletics," said Mosbach "UNI has a proud tradition of academic and athletic excellence and I look forward to building strong relationships with alumni donors and community partners to enhance opportunities for our student-athletes and programs my family and I are excited to return to the state of Iowa and to become part of the Cedar Valley community!" Mosbach was responsible for assessing the overall academic and student-athlete development programs' effectiveness and provided guidance to matters of budgeting he was a member of Arizona's NIL working group that created "Arizona Edge," which served as an educational and information platform for NIL He also managed the department's "5980 Academic Fund," which provided student-athletes additional education related benefits totaling $3 million Mosbach has also assisted the University of Arizona's development office and the U of A Foundation in securing major gifts He also has served as the secondary sport admin for Wildcat baseball and soccer having previously overseen directly the women's tennis and both men's and women's swimming and diving programs Mosbach spent six years as Arizona's Associate Athletic Director for Academics within the athletic department's comprehensive student service program Commitment to an Athlete's Total Success (C.A.T.S.) coordinated summer orientation and summer bridge programs for athletes and helped consulate with the university's Director of Student Services to help guide students through financial aid After graduating from Wartburg in 2004 as a member of the Knights' football team with a Bachelor of Arts degree in Business Administration with an emphasis in sport management UNI Athletics action can be followed all year long on social media on Facebook (UNI Athletics), X (@UNIAthletics) and on Instagram (@uniathletics). Schedules and rosters, along with the latest Panther news and information can be found online at UNIpanthers.com Community Safety Department Director To Resign Amid Tension With Cambridge Police Department From Lab to Startup: Harvard’s Office of Technology Development Paves the Way for Research Commercialization People’s Forum on Graduation Readiness Held After Vote to Eliminate MCAS 8 Takeaways From Harvard’s Task Force Reports Landscape architect Catherine Mosbach spoke about considering climate and local culture while designing landscapes at a talk hosted by the Harvard Graduate School of Design Tuesday who founded the award-winning Paris design firm Mosbach Paysagistes shared architectural drawings for five projects she pitched in architecture competitions and walked through her design process for each Mosbach focused on three different climates for landscapes she designed and how each environment shaped her approach to planning the landscapes Mosbach explained how she harnesses moisture in arid desert environments She said she implements a process of catching the humidity present in the air to support local vegetation and “bring back life” to her project sites Mosbach said she works with the seasonal changes in precipitation in continental climates To account for flooding caused by melting snow in Switzerland Mosbach looked at a water-regulation system commonly used in the Alpines The method incorporates channels surrounding the building that become rivers during the spring and prevent flooding Mosbach also talked about designing buildings in tropical regions One particular project in Taiwan included design elements such as water channels and ponds meant to capture and redirect rainwater it’s also 80% of the park full of water,” Mosbach said Mosbach said she designs landscapes to interact with surrounding buildings She showcased her designs for the University of Zurich where vegetated terraces were built into each floor of the building this was an effort to integrate buildings with their surrounding landscapes “We bring an alternative way to introduce a dialectic between the architecture and landscape,” Mosbach said Mosbach also spoke about the importance of familiarizing herself with local culture and people of the sites where she works “You need to spend time to observe and to understand the feeling and everything,” Mosbach said Mosbach also emphasized that she feels overjoyed when she sees local beneficiaries enjoying her landscapes “I am really really happy when I see the behavior of the people in the places I grow,” Mosbach said That’s the best gift a landscaper can get back.” Want to keep up with breaking news? Subscribe to our email newsletter. and the diversity of microclimates allows for use during the 12 months of the year and at any time of the day You'll now receive updates based on what you follow Personalize your stream and start following your favorite authors If you have done all of this and still can't find the email WEBSTER CITY GIRLS TRACK RETURNING LETTERWINNERS are front row (left to right): Bonnie Nohrenberg the Webster City girls track program welcomes back one returning state qualifier Lynx head coach Tyler Mosbach hopes junior Cloe Savitski will have plenty of company Mosbach likes the makeup of his squad for 2024 as Webster City brings balance and competitiveness to the table in preparation for an indoor meet at Clear Lake on Thursday “We have four returning seniors and quite a few returning letterwinners,” Mosbach said “All of them understand the expectations of the program and will be instrumental in teaching the new athletes those expectations “Cloe is our only returning state qualifier from last year but we hope to get more girls there this May.” Savitski reached the Class 3A 100-meter dash field in Des Moines as an underclassman senior Adeline Tesdahl and sophomore Lydia VanDeer were regular placewinners in meets a season ago Mosbach feels like he has the right combinations in mind for the Lynx to show vast improvement across the board “I think our biggest strengths will come in some of the relays,” Mosbach said “We don’t have that one person who is going to show up and score us 40 points but we do have a lot of girls who will give everything they can to help their teammates succeed “I look forward to that continuing and more girls following in line as the season progresses.” An offseason of work — and an influx of freshman talent — will help “We have a lot of freshmen and new faces on the team,” Mosbach said “Many are multi-sport athletes and came into the season ready “We also had most of the girls with us this winter weightlifting and speed training I would expect these girls will help us to have a well-rounded lineup giving us the ability to be more competitive at meets.” the mental aspect of growth is as important as the physical in Mosbach’s eyes “It’s about (having) an overall competitive mindset,” Mosbach said “Competition is the most important contributor to improvement It doesn’t matter if that competition is occurring in meets or in practice The girls need to strive to beat the person next to them in every rep or every event “It’s something the other coaches and I have been preaching in practice Some of the new faces have really helped us there.” Mosbach knows his squad’s current form is vastly different than what he will see in less than two months — and hopefully our ultimate goal is to get as many girls as we can down to Drake for the state meet,” Mosbach said what I really want for the team is to have as many personal records and season bests as possible that is the only thing we have any control over If we continue to get better week after week Tateelina Vankam and Ellie Weinschenk are the seniors on this spring’s squad junior Alondra Johnson and sophomore Jakaela Cherry are the other returning letterwinners March 21 — at North Indoor Indoor Championships in Clear Lake April 2 — at Southeast Valley; 4 — at Eagle Grove; 9 — at Algona; 25-27 — Drake Relays; 29 — at Iowa Falls-Alden; 29 — at Iowa Falls-Alden GARNER — The Webster City girls soccer team made it two in a row here last Thursday LLC | https://www.freemanjournal.net | P.O control and learning to let go when designing for living systems Foreground: You have spoken about giving up some control when designing landscapes but your built work shows extremely precise and beautiful design whether in specifying materials and defining and locating elements or managing the various stages of the work What is the relationship between ‘control’ and ‘responsibility’ for you Catherine Mosbach: Control is linked to the way you engage in a dialogue with a site The term is not best adapted to describe the process – let’s use the word ‘design’ a diverse collective of users want different things and that requires us to resolve questions of services That means we use our expertise and knowledge to draw together the scope of many possibilities to create a unique place Then it is a question of what we know and play with today We do not know this so precisely but we aim to leave open a creative dialogue between present and future Landscape – like an animal or vegetal body – is always changing fixed ‘place’ is quite a strange way to translate our position on the earth To make a dialogue between what you know and what you do not know you need to introduce some conditions or rules We cannot speak together without an alphabet and rules but that doesn’t mean the ‘text’ of our conversation is locked or predetermined Design is a language and it should be in continuous dialogue – in landscape design – with life happenings “That’s the goal of a public space: bringing together many kinds of life in ways that engage our creativity and humanity” – Catherine Mosbach FG: You seem delighted when users – human and animals – take ownership of spaces and behave in unexpected ways in your projects What judgements do you make about unplanned activities Is this a calculated ‘balance’ or an unspoken subversion or something less definable or conscious in the design process CM: We – I – work for public space and for many generations I am a human with a singular understanding of a context and how a question addresses a program five different landscape architects may propose different translations of the context and different answers to questions I understand design as a dialogue between me and the site And that discussion is addressed to a public inhabitant For me in landscape – like in life – a ‘piece’ or project has many functions and not only one functions must already account for changes of season and longer timelines of growth When I encounter people from different ages and cultures I am inspired and I imagine the larger public It is a great gift to have the capacity to see and imagine as another like all artistic work that encounters a public That’s the goal of a public space: bringing together many kinds of life in ways that engage our creativity and humanity But a dialogue doesn’t mean talking over the host The origin and point of the conversation is lost with overlapping and competing voices the opening of an evolving movie where many lives – animals humans – will play parts together after being introduced FG: In the face of a changing climate and its unpredictable yet inevitable impacts will a flexible approach to the types of control we have of landscapes become more necessary or perhaps inevitable Has ‘control’ tended to create a paradigm that has contributed to pollution and global warming CM: Control isn’t necessarily helping worthy contributions We are living with knowledge that is always evolving The knowledge held by different disciplines is continuous and exponential Great achievements rarely happen within the span of a human life but grow on the achievements of others over many years and even generations It is linked to the tools we use both in micro life (biology We all live with the state of knowledge and the tools we have in our own times We understand that the global behaviour of all populations together won’t fit with the needs of other life on earth So we need to introduce a more subtle dialogue between our ‘human’ system and other ‘systems’ But we can only use the tools from our timeline – with enough flexibility to embrace what we do not know – and survive by being as efficient as possible If we only had to efficiently apply what we know If no human thought or effort or sensibility is necessary Today a dominant movement in capitals and countries is to ask all inhabitants to express their ideas to contribute to the program –- that is direct democratic decision-making It assumes that it is not necessary to have experience and knowledge to make decisions It supposes everyone can equally read and work through all the parameters of a problem; that everyone can do it It actually happens when nobody wants to take responsibility for a direction It is a way to say that expertise is not valid and everyone has an equally legitimate opinion The result of this is what we see on social media and in the fragility of democracy Fraternity was a call to control human wildness and open a dialogue to bring together individuals and communities to make decisions affecting us all Today we need to do that with other life as well as each other FG: Your work is renowned for the delightful and profound ways it engages with temporality and continuing change Can you explain how you work with the perception of time in gardens and landscapes you need to understand the largest processes of our environment’s evolution to anticipate further stages Leaving the system open is a flexible way to accept unexpected events although if you let the work be completely open to unexpected phenomenon you actually have no project and no writing or sensitive translation That’s the limit of ecology: just trying to reproduce what is I typically introduce several layers of rhythms An urban landscape needs to be interesting for all seasons mixing young trees with mature trees so that there is always a transition like in real life A project should consider each season and connect us to the real facts of the landscape not a kind of final product to be presented as merchandise is that a project can take from five to eight years But that is only the beginning and the initial conception has to be followed by maintenance What is fabulous in our practice – which is the opposite with architecture – is that landscape architecture can continue to work with inherited projects over time But that supposes the translation is respected – like in Versailles for the last 400 years – and this is very rare and problematic What types of knowledge can or should designed landscapes ‘teach’ CM: The mayor requested we look at Steiner philosophy experiments with alternative ways to learn Basically it means more interaction between teacher and student more care about bodies that is not only about appearance but about complex interactions with the world What that actually meant for the park was that the architect’s climatic devices focused on only one function per installation and for a small group of users Taiwan’s Phases Shift Park is polysemic – it has many meanings and offers many ways of approaching and appreciating change rather than just through immediate climate mitigation or manipulation That makes it very powerful and efficient for future inhabitants Outdoor life is very unusual in Taiwan because of a lack of comfort So testing and learning about your body with and compared to other bodies is a way to open your mind to other things you do not know So the project of a park is a social and political investment working both for individuals and for a collective capacity to evolve and create together And this is the key lesson for sharing and surviving and will never be able to create mountains or seas we need to find a subterfuge to make as if we can” – Catherine Mosbach FG: You have spoken of ‘the thickness of the nourishing factor’ of design Can you tell us more about what ‘nourishing’ means CM: My background in biology, physics and chemistry covers the parameters of life on earth. Since I developed Pages Paysages I have met many inspiring people who nourish my curiosity In occidental culture we are used to seeing and evaluating only the results of investigations as fixed conclusions But in Asian cultures what we see is never an end to create a life process you need to understand where the result is coming from The request in Bordeaux for the Botanic Gardens was ambitious and I did not want to reproduce the invisible on the surface like a ‘museographer’ showing stable object ‘fossils’ Real life is much more powerful than our brains can imagine and teach people to evolve in real life without being afraid of what is hidden Bordeaux is much more powerful than what I could have imagined But then there are other weaknesses – life is never perfect “I like to dream – then invite guests to dream too in their everyday life!” – Catherine Mosbach FG: Your projects render unseen and subsurface things visible – making ‘representations in the literal sense’ as you say of Bordeaux Botanic Gardens Can you explain how miniaturisation and shifts in scale have become a useful tactic for you I have done three small pieces – Quebec 250sqm Ulsan 500sqm – and all look like they are endless And I like to dream – then invite guests to dream too in their everyday life “A representation is what the name says – a translation FG: In their enjoyment of the Botanic Gardens some people did not notice that it was nature recomposed but thought it was merely part of a landscape transported to the site Some might think it is desirable for a project to seamlessly replicate or be mistaken for nature What is important to you about interpretation and its distinction from imitation as if in the occident we cannot have another way to think aside from some mainstream accepted and familiar manner of organising and ornamenting space Sanaa did not ask me and I did not develop a design to follow Japanese style I want to make possibilities seem as large as possible so as to open the mind to imaginative exploration Without it you cannot represent twelve environmental landscapes in two hectares that means we need to use techniques to keep the scale through timelines because one large tree in a particular spot on the earth has nothing to do with a forest representing an environment A representation is what the name says – a translation Would a lover give an imitation gold jewel to their beloved I just try to be inspired with the power of what exists to look beyond what we know because our life has a limited span and little time to discover its  many ‘treasures’ so we need to train beforehand – a submersive experiment so as not to be surprised News and analysis on cities, places and the people who make them delivered direct to your inbox FLORIDA – If the Gulf Coast High girls basketball team is facing an in-county opponent Gulf Coast won the Class 6A-District 12 championship Friday while extending a lengthy winning streak their 77th straight victory against a Collier County opponent dating back to January 2016 they needed a third quarter explosion to secure their third district title in four years Gulf Coast (19-4) went on an 18-0 run spanning the second and third quarters to put the game out of reach I'm so proud of how far we came," Sharks junior Lilly Fultz said "We stuck with it even though the game was a little close We stuck with it and we ended up winning a trophy." After leading 23-16 late in the second quarter Gulf Coast scored the first 11 points of the third quarter and coasted from there Sophia Pasquerello scored 13 points for the Sharks and hit three 3-pointers Gulf Coast finished with eight threes as a team Junior Meloday Charlton led Palmetto Ridge with 12 points Palmetto Ridge finished the season 10-10 and got to the district final in the first season under new coach Ron Mosbach and they had just one winning season in that span Addie Osborne added 11 points for Gulf Coast who will host a regional quarterfinal on Wednesday the top-seeded Celtics (22-3) edged past Seacrest (21-5) Addilyn Bailey gave the Stingrays the lead with a 3-pointer with 90 seconds left but Neumann's Sophia McCarthy made the go-ahead bucket with less than a minute to go It was the third straight district title for Neumann Seacrest was led by Dragana Secpanovic with 15 points Gulf Coast HEAT 18: After winning 95-7 in the district semifinals the Sentinels (21-3) had almost as easy of a time in the finals ECS was up 38-2 after the first quarter and led 53-10 at the half.Kellisia Grant had 14 points to lead the Sentinels Canterbury 32: Cameron O'Halloran led the way for the Vikings scoring 18 points Freshman Caroline Ward added 17 to help Verot hold off Canterbury Kaya Langford scored 12 points for Canterbury including 10 in the fourth quarter to keep the Cougars within striking distance Gateway 56 Aubrey Rogers 37: In a battle of new programs the Eagles won their first district title in their third season of existence jumping out to an 11-3 lead and never looking back The Eagles were up 17-7 after the first period This was a battle of the newest school in Lee County (Gateway) vs Angelina Insana scored 14 to lead the Patriots Cypress Lake 34: The Pirates led the entire way scoring the game's first nine points on their way to a fourth consecutive district title Port Charlotte led by 10 after the first quarter and 25 after the third.Hailey Cohen had a double-double Mariah Epting added 14 points and 9 steals Lilly Witz scored 14 and was the only Panthers player in double figures Barron Collier 34: The hottest team in Southwest Florida knocked off the most successful program in Collier County as Lely won its first district title in five years had won 13 district championships the past 14 seasons The Trojans now have won 13 straight games and are headed to the playoffs after three straight losing seasons Lely trailed 24-19 at halftime but locked down defensively in the second half Sofiia Kiritsa led Lely (18-8) with 19 points Ida Baker 13: The Tarpons (15-5) raced out to a 25-7 lead at the end of one then did even better in the second quarter Then outscored Baker 25-0 to lead 50-7 at halftime and the entire second half was played under a running clock.Acelia Beals put up 22 points for Charlotte McKenzie Cole was Baker's leading scorer with six points As reported earlier flash flooding affected parts of France and Belgium after more than 50mm of rain fell in around 1 hour on 02 June 2021 Stormy weather later affected areas of the Netherlands where some minor flash flooding was reported in Deelen As much as 42.9 mm of rain fell in about 1 hour in Deelen on 03 June 2021 Heavy rain of over 60 mm was reported in Recken op der Mess saw more than 40 mm of rain in 1 hour on 03 June More than 180,000 lightning strikes were reported on 04 June Voelkersleier in Bavaria recorded 59.2 mm of rain in 24 hours to 06 June A serious road accident occurred in Düsseldorf after a young cyclist was hit by lightning One person died and 3 were injured after weather-related accident at a construction site in Stuttgart Flash flooding was reported in parts of Thuringia The Fire Service was called out to hundreds of interventions mostly to attend to flooded streets and cellars Some of the most severe flooding struck in Mosbach where local streams overflowed sweeping away vehicles and prompting at least 3 rescues The Civil Protection Directorate in Croatia reported that severe flash flooding affected areas of Požega-Slavonia Zagreb and Bjelovar-Bilogora counties 06 to 07 June As much as 70 mm of rain had fallen in some locations The worst affected areas were the town of Požega and the settlements of Drškovci Vidovci and Dervišaga in Požega-Slavonia County Civil Protection said homes and roads were flooded and small landslides also reported In the settlement of Vidovci the electrical network was turned off due to the danger for firefighters Richard Davies is the founder of floodlist.com and reports on flooding news Cookies | Privacy | Contacts © Copyright 2025 FloodList Volume 10 - 2017 | https://doi.org/10.3389/fnmol.2017.00433 This article is part of the Research TopicBiology of Lipids in the Control of the Motor Function, in Health and DiseasesView all 13 articles Lipid metabolism is drastically dysregulated in amyotrophic lateral sclerosis and impacts prognosis of patients Animal models recapitulate alterations in the energy metabolism including hypermetabolism and severe loss of adipose tissue To gain insight into the molecular mechanisms underlying disease progression in amyotrophic lateral sclerosis we have performed RNA-sequencing and lipidomic profiling in spinal cord of symptomatic SOD1G86R mice Spinal transcriptome of SOD1G86R mice was characterized by differential expression of genes related to immune system Hypothesis-driven identification of metabolites showed that lipids and phosphatidylcholine(o-22:1/20:4) showed profound altered levels A correlation between disease severity and gene expression or metabolite levels was found for sphingosine Joint-analysis revealed a significant enrichment of glycosphingolipid metabolism in SOD1G86R mice and lactosylceramide and the overexpression of genes involved in their recycling in the lysosome A drug-gene interaction database was interrogated to identify potential drugs able to modulate the dysregulated genes from the signaling pathway Our results suggest that complex lipids are pivotally changed during the first phase of motor symptoms in an animal model of amyotrophic lateral sclerosis We present for the first time an integrated analysis of RNA-sequencing and lipidomic data from the spinal cord of symptomatic SOD1G86R mice These results allow an unbiased vision of the metabolic changes in the course of ALS and pinpoint specific pathways which are pathologically important Mice were maintained in our animal facility at 23°C with a 12 h light/dark cycle Mice had access to water and to regular A04 rodent chow ad libitum Body mass and muscle strength were analyzed on a daily basis to access disease onset For muscle strength measurements (mean of three tests a mouse was placed on grid where they spontaneously gripped it with their four paws They were then gently pulled back by the experimenter until they released the grid (mean of three tests A strength-meter recorded the peak grip strength of the mouse Age and litter matched non-transgenic female mice served as control mice were sacrificed by intraperitoneal injection with ketamine chlorohydrate (100 mg/kg) and xylazine (5 mg/kg) and intracardially perfused with PBS at 4°C Lumbar spinal cords were quickly dissected and kept at −80°C until further analysis After normalization and rlog transformation and PCA (principal component analysis) was performed with the indicated subset of genes Validation of gene expression was assessed by qPCR with a CFX96 using SYBR green Supermix reagent (BioRad) Relative quantification of each gene was determined using the Biorad software and normalized to reference genes (Pol2 Primers sequences are provided in Supplementary Table 1 Comparison between groups was studied with student's t-test and p-value < 0.05 were considered significant (PRISM 6.0b Data are expressed as the mean ± SEM RNA-sequencing data are available at the Gene Expression Omnibus (GEO) database repository Lipid extraction and UPLC/TOF-MS (ultra performance liquid chromatography/time-of-flight mass spectrometry) was performed as previously described (Henriques et al., 2015b) spinal cord samples from 95 days old mice were homogenized in precooled methanol (n = 7 Chloroform was added and after centrifugation organic phase was collected and evaporated Residues were solubilized with acetonitrile/isopropanol Chromatography was performed on an Acquity UPLC system using an Acquity BEH C18 column The chromatographic system was coupled to a LCT Synapt G2S mass spectrometer (Waters Corporation) equipped with an electrospray source operating in positive or negative ion mode with a lockspray interface for accurate mass measurements Switzerland) was use to normalize metabolomics data Data with retention times between 3 and 15 min with a mass range between 300 and 1,000 and peak intensity distinct from zero were retained and normalized to fresh tissue mass Data were submitted to PARETO transformation before statistical analysis Statistical analysis for differential lipid metabolite level was performed by using supervised multivariate orthogonal partial least-squares discriminant analysis (OPLS-DA) A difference was considered significant when the corresponding variable |correlation coefficient| was greated than or egal to 0.7 according to the p(corr) coordinate in the S-plot built after the OPLS-DA model Molecular features with significant changes were associated with theoretically identified metabolites based on their atomic mass (m/z) Together with lipid extraction from samples and liquid chromatography gradient the brut formula associated with the exact mass-over-ionization-state ratio allowed tentative lipid identification Pathways represented by at least 3 genes and 3 lipid metabolites and with q-value for enrichment below 0.05 were considered as significantly altered Transcriptomic dysregulation in the spinal cord of SOD1G86R mice Expression level of selected genes as determined by RNA-sequencing and qPCR Validation of differential gene expression by qPCR and compared to the fold change determined by RNA sequencing qPCR values were normalized to the mean of the respective WT group qPCR data is presented as mean ± standard variation of the mean These results suggest that these pathways are strongly impacted in symptomatic SOD1G86R mice Multivariate analysis showing samples distribution based on different subset of genes Unsupervised principal component analysis performed with genes isolated from “lysosome” (A) “external side of membrane” (B) and “membrane raft” (C) gene ontology terms Other subclasses of dysregulated lipids included di- and triglycerides Saturation and unsaturation of acyl chains directly influence biophysical properties of complex lipids with repercussion on membrane fluidity down-regulated sphingolipid metabolites mainly quasi-exclusively contained saturated and monounsaturated fatty acids This suggests that sphingolipids having polyunsaturated acyl chains were not significantly affected in SOD1G86R mice This difference in acyl chains could reflect enzymatic activities involved in the synthesis transformation or degradation of fatty acids and lipids Dysregulation of sphingolipid metabolites in the spinal cord of SOD1G86R mice Dysregulation of phospholipids metabolites in the spinal cord of SOD1G86R mice When considering negative ionization, similar results were obtained. HMDB identification was attributed to 165 lipids metabolites showing significantly altered levels in SOD1G86R mice. A majority of lipid metabolites were down-regulated. Main lipid subclasses were fatty acids, phospholipids (mainly phosphatidylcholine), sphingolipids (e.g., ceramide), and tri-, di-, or mono-acylglycerides (Table 4) several significantly altered lipid metabolites received an identical identification after both positive and negative ionizations These lipids were sphingolipids [Cer(d18:1/22:1); Cer-P(d18:1/26:1); Cer(d18:1/18:0)] triglycerides [TG(18:4/15:0/18:4); TG(14:1/20:5/14:1)] or from other lipid subclasses [5-Hydroxy-7-methoxy-2-tritriacontylchromone; N-oleoylethanolamine; Randilongin; Persenone a] Dysregulation of lipid metabolites in the spinal cord of SOD1G86R mice Overlapping genes are those coding for lysosomal proteins involved in the recycling of glycosphingolipids (e.g. an enzyme involved in the degradation of sphingosine-1-phosphate Overlapping lipid metabolites were ceramide mono-hexosylceramide (glucosylceramide or galactosylceramide) were not detected as significantly enriched Integrated pathway analysis of spinal transcriptomic and lipidomic data of SOD1G86R mice Ugt8a codes for the galactosylceramide synthase SGPP2 and SPHK1 are two enzymes involved respectively in the synthesis and degradation of sphingosine-1-phosphate Regulation and interaction of genes related to glycosphingolipid pathway performed with genes from the KEGG pathway “sphingolipid metabolism pathway,” showing distribution of SOD1G86R and wild type samples (blue (B) Validation by qPCR of differential expression level of genes from the sphingolipid pathway previously found dysregulated by RNAseq (C–G) Pearson correlations between muscle strength and gene expression levels of Ugt8a (C) and metabolite levels of sphingosine (F) and ceramide(d18:1/26:0) (G) (H) Schematic representation of the main dysregulations related to glycosphingolipids Green boxes represent metabolites and blue boxes refer to genes In red are given genes and metabolites showing either altered levels or a correlation with disease severity Gene abbreviation follows the HUGO Gene Nomenclature Committee-approved gene nomenclature Data are presented as mean ± standard variation of the mean not significant; #p < 0.1; *p < 0.05; **p < 0.01 are bioactive molecules able to module cell response to stress KEGG identified “sphingolipid signaling pathway” as dysregulated suggesting that altered levels of sphingolipids in ALS could translate into changes in cell signaling DGIdb database was used to search for potential pharmaceutical modulators of the “sphingolipid signaling pathway” using genes identified as dysregulated in this pathway (Wagner et al., 2016) including 10 negative modulators of TNF-a signaling and 5 modulators of sphingosine-1-phosphate 6 of them are currently investigated or were tested at preclinical and/or clinical level in ALS (Pyrimethamine These findings indicate that the metabolism of sphingolipid shows dysregulation at gene and metabolites levels in the spinal cord of symptomatic SOD1G86R mice ALS is a fatal condition characterized by degeneration of motor neurons Several lines of evidence suggest that gene expression and lipid metabolism are differentially regulated in ALS and could contribute to disease progression we report a joint analysis of lipidomic and transcriptomic changes in the spinal cord of symptomatic SOD1G86R mice the up-regulation of Runx1 in the spinal tissue of SOD1G86R mice could represent a compensatory neuroprotective mechanism Our data also shows that sphingolipids and phospholipids are under-represented These lipid classes are key components of the cell membrane Unsaturation impacts membrane fluidity and low proportion in monounsaturated acyl chains leads to higher stiffness Disturbance in membrane fluidity is known to impair cellular function such as cell signaling common dysregulations in the spinal cord of an animal model of ALS Our analysis identified sphingolipid metabolism as the most dysregulated biological pathway Sphingolipids are complex lipids derived from sphingosine Similarly, ceramide-phosphate promotes cell survival through the PI3-K/PKB pathway (Gómez-Muñoz et al., 2005; Gómez-Muñoz, 2006) two genes involved in the synthesis of ceramide-phosphate and sphingosine-1-phosphate could be a protective mechanism for counteracting neurodegeneration Our results reinforce the critical role of complex lipids Dysregulated glycosphingolipid metabolism could therefore participate in the progression of ALS symptoms in SOD1G86R mice remain predictive tools for studying pathological processes of ALS Our study highlights dysregulations of sphingolipid metabolism in SOD1G86R mice at symptomatic disease stage. As bioactive molecules, sphingolipids interplay with many cell pathways that could be targeted with drug candidates. Pharmacological modulators targeting the metabolism of sphingolipids exist and have been proposed for the treatment of human diseases (Canals et al., 2011) In silico analysis has identified fingolimod and pyrimethamine as tentative modulators for ALS These two drugs modulate sphingosine-1-phosphate signaling and have been already tested in ALS Fingolimod improved motor functions and survival of SOD1G93A mice and has been given to ALS patients in a clinical phase 2a safety study Pyrimethamine has been shown to successfully lower mutant SOD1 proteins in the cerebrospinal fluid of ALS patients carrying SOD1 mutations Other potential drug candidates have been tentatively identified and most of them have not been investigated in ALS The facts that our data-driven analysis identified pharmacological compounds already tested in ALS reinforce the relevance of our results spinal cord of SOD1G86R mice present with profound dysregulations at transcriptomic and metabolomic levels that point to sphingolipids Our results complement existing data on dysregulation of complex lipids in ALS and could open new therapeutic strategies linked to sphingolipids and BW: Conceived and designed the experiments; AH and J-PL: Analysis and interpretation of the data; VC discussed and approved the final version of the manuscript This work was supported by the European Community's Health Seventh Framework Programme under grant agreement No “Association André combat la SLA” and “Association pour la Recherche et le Développement de Moyens de Lutte contre les Maladies Neurodégénératives” (AREMANE) RNA-sequencing was performed by the IGBMC GenomEast platform a member of the “France Génomique” consortium (ANR-10-INBS-0009) AH and MS are employees of Spedding Research Solutions SAS and CB-N are employees of Les Laboratoires Servier The other 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 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an Animal Model of Amyotrophic Lateral Sclerosis Received: 31 October 2017; Accepted: 15 December 2017; Published: 04 January 2018 Copyright © 2018 Henriques, Croixmarie, Bouscary, Mosbach, Keime, Boursier-Neyret, Walter, Spedding and Loeffler. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY) distribution or reproduction in other forums is permitted provided the original author(s) or licensor are credited and that the original publication in this journal is cited in accordance with accepted academic practice distribution or reproduction is permitted which does not comply with these terms *Correspondence: Alexandre Henriques, aGVucmlxdWVzLmFAZ214LmNvbQ== Jean-Philippe Loeffler, bG9lZmZsZXJAdW5pc3RyYS5mcg== Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. 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. 94% of researchers rate our articles as excellent or goodLearn more about the work of our research integrity team to safeguard the quality of each article we publish. WEBSTER CITY GIRLS TRACK RETURNING LETTERWINNERS are, front row (left to right): Adysen Poppen, Charlee Olson, Cloe Savistiki, Izzy Grossoehme and Lydia VanDeer. Back: Amanda Lahr, Jakaela Cherry, Avery Poppen and Tristyn Wright. (DFJ photo by Britt Kudla) The Webster City girls track and field team returns a pair of individual Top-20 state finishers this spring alongside several underclassmen looking to make their own mark on the program. Sophomore Izzy Grossoehme was 14th in the Class 3A 400-meter dash a season ago, while senior Adysen Poppen placed 19th in the shot put. Classmate Cloe Savitski and Grossoehme are two legs from the state 4×400-meter relay unit back for head coach Tyler Mosbach. “We’ve been flexible with our practice schedule and the weather,” Mosbach said. “But I’ve really relied on Cloe and Izzy, pushing them into being leaders for the team. The younger girls really look up to them and they have done a great job of showing them all the little stuff so us coaches don’t have to always be watching that.” Poppen, who suffered a knee injury in the fall, continues to work her way back into competing. “It’s just great that Adysen is out and can give it a go,” Mosbach said. “Whatever she can do will be great to see. I’m just happy she can experience this season.” Along with Savitski and Poppen, the senior class includes Lydia VanDeer. Hailey Brim, Jakaela Cherry and Cindy Gomez Lopez are the juniors. “We as coaches for girls sports at Webster City have really tried to come together to incorporate everything with one another for the kids,” Mosbach said. “That includes lifting, speed training and working to build a strong culture of girls sports at the school. “We want them to know what we expect out of them in regards to working. There is no offseason and you always have to continue to get better. As we create and grow that culture, it’s going to lead to a bright future.” Sophomores are Perla Andrade-Murillo, Lilly Haney, Kali Hartnett, Shamyla Millner, Charlee Olson, Avery Poppen, Elexious Scism, Kaitlynn Van Demark, Tristyn Wright and Grossoehme. Freshmen out are Rubi Andrade-Murillo, Gracie Bishop, Mckenna Buakham, Evie Davis, Kali Hartnett, Greta Kenville, Zoie MacRunnel, Haedyn Soesbe, Briley VanDeer and Isabel Weinschenk. Mosbach will be assisted by Clint Howard and Brad VanDeer this spring. Webster City heads to Clear Lake on Thursday for the North Iowa Indoor Championships, before opening the outdoor season on April 7 in Gowrie. Seniors — Adysen Poppen, Cloe Savitski, Lydia VanDeer. Juniors — Hailey Brim, Jakaela Cherry, Cindy Gomez Lopez. Sophomores — Perla Andrade-Murillo, Izzy Grossoehme, Lilly Haney, Kali Hartnett, Shamyla Millner, Charlee Olson, Avery Poppen, Elexious Scism, Kaitlynn Van Demark, Tristyn Wright. Freshmen — Rubi Andrade-Murillo, Gracie Bishop, Mckenna Buakham, Evie Davis, Kali Hartnett, Greta Kenville, Zoie MacRunnel, Haedyn Soesbe, Briley VanDeer, Isabel Weinschenk. March 27 — at North Iowa Indoor Championships in Clear Lake. April 7 — at Southeast Valley; 10 — at Eagle Grove; 15 — at Algona; 21 — at Greene County; 24 — host Lynx Relays; 28 — at Humboldt. May 1 — at Boone; 5 — at Iowa Falls-Alden; 8 — at NCC Championships in Iowa Falls. JEWELL — The South Hamilton girls golf team placed third here last Thursday behind Roland-Story and West Marshall ... MANSON — The South Hamilton boys and girls track and field teams were in action here last Thursday night at the ... EAGLE GROVE — Luke Martensen had the low round as the South Hamilton boys golf team claimed a triangular here ... Copyright © 2025 Ogden Newspapers of Iowa, LLC | https://www.freemanjournal.net | P.O. Box 490, Webster City, IA 50595 | Volume 8 - 2017 | https://doi.org/10.3389/fmicb.2017.02361 Crop protection anilinopyrimidine (AP) fungicides were introduced more than 20 years ago for the control of a range of diseases caused by ascomycete plant pathogens and in particular for the control of gray mold caused by Botrytis cinerea Although early mode of action studies suggested an inhibition of methionine biosynthesis the molecular target of this class of fungicides was never fully clarified Despite AP-specific resistance having been described in B cinerea field isolates and in multiple other targeted species the underlying resistance mechanisms were unknown It was therefore expected that the genetic characterization of resistance mechanisms would permit the identification of the molecular target of these fungicides In order to explore the widest range of possible resistance mechanisms cinerea UV laboratory mutants were generated and the mutations conferring resistance were determined by combining whole-genome sequencing and reverse genetics Genetic mapping from a cross between a resistant field isolate and a sensitive reference isolate was used in parallel and led to the identification of an additional molecular determinant not found from the characterized UV mutant collection these two approaches enabled the characterization of an unrivaled diversity of resistance mechanisms we report the elucidation of resistance-conferring mutations within nine individual genes two of which are responsible for almost all instances of AP resistance in the field All identified resistance-conferring genes encode proteins that are involved in mitochondrial processes suggesting that APs primarily target the mitochondria The functions of these genes and their possible interactions are discussed in the context of the potential mode of action for this important class of fungicides cyprodinil-containing products are used for the control of cereal diseases such as powdery mildew (Blumeria graminis f.sp tritici) and Parastagonospora nodorum on wheat and scald on barley caused by Oculimacula spp. Pyrenophora teres and Rhynchosporium commune Pyrimethanil-containing products are used for the control of Ascochyta spp In this paper we took advantage of our contribution to the recent assembly of a gapless B. cinerea genome (Van Kan et al., 2017) to (i) determine resistance mechanisms in AP-resistant UV lab mutants using a whole-genome sequencing approach (ii) determine the most frequent resistance mechanism in AniR1 field samples using a sequenced mapping population that we initially generated for producing a high density linkage map of Botrytis and (iii) used these complementary information sets to characterize B Our work enabled us to demonstrate a partial overlap between the two approaches and to discover an unrivaled level of complexity in terms of resistance mechanisms We demonstrated the individual involvement of mutations within at least eight separate genes from our UV mutagenesis approach and characterized a wide range of resistance-conferring mutations that occur in two distinct genes in field isolates All resistance-conferring mutations affected nuclearly-encoded mitochondrial proteins supporting a mitochondrial mode of action for AP fungicides We discuss known and proposed interactions between these genes present possible explanations for the observed phenotypes and hypothesize on the likely target of this important class of fungicides The fully sequenced anilinopyrimidine (AP) sensitive B. cinerea reference strain B05.10 was used as genetic background for UV mutagenesis screening and for all reverse genetic confirmation of cyprodinil (CDL) resistance-conferring mutations. The two AP-resistant field isolates 09Bc11 (Van Kan et al., 2017) and BAR633 (kindly provided by Stefania Pollastro Italy) were used for genetic crossing experiments which correspond to either mixed populations of up to three genotypes or single isolates as determined by subsequent genotyping methods were mostly collected from trial sites and obtained by collection of conidia from the surface of infected plant tissues using sterile cotton swabs Samples were propagated and phenotypically tested at BIOtransfer laboratory (Montreuil Samples isolated in 2014 originate from grape (n = 303 as shown in Table S1 excluding samples displaying multiple genotypes cinerea samples for which Sanger sequencing was performed are listed in Table S2 with the respective appropriate CDL supplement Cultures on agar plates were incubated at 20°C the rest after autoclaving); agar 25 g·L−1] at 25°C for 4 days before transfer to GEA plates for spore production (19°C Mycelium material for protoplast generation was produced in NY liquid medium (yeast extract 200 mg·L−1; malt extract 20 g·L−1; pH 5.5) GG medium (KH2PO4 1.5 g·L−1; MgSO4 ·7H2O 750 mg·L−1; gelatin from porcine skin 4 g·L−1; glucose 4 g·L−1) was used for liquid culture dose-response tests All oligonucleotides were purchased from Microsynth AG (Balgach Genomic DNA used as template for polymerase chain reactions was prepared using the MagAttract 96 DNA Plant Core Kit (QIAGEN based on freeze-dried sporulating mycelium of the respective field samples crossing progeny isolates (both harvested from GEA plates) or transformants (both from Vogel's minimal agar with the respective appropriate CDL supplement) Each PCR was performed according to the conditions recommended by the respective manufacturer of the polymerase and using ~50 mg of DNA template per reaction PCR primers used to amplify sequences for Sanger sequencing or reverse genetic experiments (transformations) are listed in Table S3 PCR products for reverse genetic validation of resistance-related mutations and for Sanger sequencing were amplified with Thermo Scientific Phusion Hot Start II High-Fidelity DNA Polymerase (ThermoFisher Scientific For the long products required to map the Bcpos5 locus DNA Sanger sequencing was done at Microsynth AG or internally (Applied Biosystems 3130 Genetic Analyzer) Oligonucleotides designed for pyrosequencing assays together with other assay-related information PCR products for pyrosequencing assays were amplified from DNA templates of field samples (of which some represented mixed populations) or crossing progeny isolates with GoTaq® G2 Hot Start Polymerase (Promega Pyrosequencing was performed on a PyroMark Q96 ID (Biotage/QIAGEN) according to the protocols provided by the manufacturer (Allele Quantification and SNP Genotyping assays in the PyroMark Assay Design software v2.0) Chemicals used for in vitro fungicide sensitivity and cross-resistance assays were purchased from Sigma-Aldrich: Cyprodinil (CDL Stock solutions of antifungal compounds were prepared at 10 g·L−1 in dimethyl sulfoxide (DMSO) cinerea monitoring field samples (Table S1) and of the mapping population from the cross between 09Bc11 and BAR633 (see section Mapping of Resistance Loci in Field Isolates and Phenotypic Characterization) were performed at BIOtransfer: Conidia were picked with an agar plug from sporulating cultures on GEA and suspended in sterile distilled water The spore suspension was then adjusted to 2·105 conidia per ml in GG medium Technical active ingredients dissolved in DMSO were used to prepare adapted concentration ranges in sterile distilled water Each well of a 96-well microtiter plate was filled with 100 μL of 2x concentrated fungicide solution and 100 μL of conidia suspension The final concentration of DMSO in each well including the solvent control was 1% (v/v) Final dose ranges used for monitoring samples were 0 Concentrations used for phenotyping the 09Bc11 x BAR633 mapping population ranged between 0.0017 and 100 mg·L−1 with 1:3x dilution steps cinerea sample or progeny isolate was tested in technical duplicate Microtiter plates were incubated in the dark at 19°C for 3 days before measuring optical density at 450 nm with a Biotek plate reader (ELX800 UV) Measured values were subtracted from the blanks measured directly after inoculation EC50 values were calculated using the Grafit 5 software (Erythacus Ltd.) Liquid culture cross-resistance assays were performed using B single spore isolates based on the original field samples listed in Tables S1 or transformants (see section Validation of Resistance-Related Mutations by Homologous Recombination) Spores were harvested from 7 to 10 days old cultures on GEA The spores were suspended in GG medium and adjusted to a density of 106 conidia per mL The suspensions were then diluted 1:10x with GG medium and 198 μL were distributed into each well of a 96-well microtiter plate (Costar 3370 already containing 2 μL of 100x concentrated compound solutions in DMSO resulting in a final DMSO concentration in all assays of 1% (v/v) Dilution series of compounds were prepared in ten 1:3x dilution steps starting from 10 g·L−1 stock solutions resulting in a final concentration range from 0.0017 to 100 mg·L−1 Optical density of the cultures at 600 nm was measured directly after treatment (blank) and after 3 days of incubation in a growth chamber at 20°C in the dark using the EnSpire 2300 Multilabel Reader (PerkinElmer) EC50 values were calculated using GraphPad Prism 6.0 (GraphPad Software with the function: Dose-response–inhibition response–Variable slope (four parameters) cinerea B05.10 were collected from 7 to 12 days old cultures on GEA or 2·107 spores were suspended in sterile water and spread onto Vogel's agar 9 cm Petri dishes supplemented with CDL The selective concentrations of CDL varied between 10 and 100 mg·L−1 For a detailed description of the different conditions applied see Table S5 UV mutagenesis was performed using 65 mJ·cm−2 treatment with a UV Stratalinker 2400 (Stratagene) Non-selective agar plates as well as non-UV-treated controls were included After treatment the plates were incubated for at least 10 days in the dark at 20°C Single colonies were transferred to new selective Vogel's agar plates with a similar concentration of CDL for secondary selection and left to sporulate under light Isolates were phenotypically tested and only those showing stable resistance were used for further experiments (Table S5) Thirteen resistant UV mutants and the sensitive B05.10 wild type reference were selected for whole-genome sequencing. Illumina sequencing library preps were generated as described (Van Kan et al., 2017) Sequencing was done on an Illumina HiSeq 2000 (Illumina Inc. USA) as either 54 or 100 cycle paired end runs with an average yield of 3.4 Gb per sample (range 1.4–5.4 Gb per sample) Variants were annotated with respect to effects on protein coding regions and then further filtered by requiring at least 10 reads of unique coverage a variant frequency of ≥70% in the UV mutant and of ≤10% in the reference strain Candidate mutations were assessed from the resulting subset of SNPs and small indels Table S6 lists the non-synonymous candidate mutations and the total number of high-confidence mutations obtained per UV isolate The cross between 09Bc11 (AniR1 phenotype) and B05.10, and the generation of a genetic linkage map for 70 progeny isolates (35 sensitive and 35 resistant toward CDL) have been described previously (Van Kan et al., 2017). A second cross was performed between 09Bc11 (maternal) and strain BAR633 (AniR1) according to established procedure (Faretra et al., 1988) EC50 values were determined at BIOtransfer for 169 progeny isolates using the liquid culture sensitivity test described above Pyrosequencing of individual progeny isolates was used to genotype for the presence or absence of resistance-conferring mutations within the Bcpos5 and Bcmdl1 genes (pyrosequencing assays Bcpos5-AQ-L412F and Bcmdl1-AQ-E407K Genomic regions encompassing the identified SNPs suspected to confer AP resistance were amplified from genomic DNA by PCR using Phusion Hot Start II High-Fidelity DNA Polymerase (ThermoFisher Scientific including flanking regions of at least 500 bp up- and downstream of the respective SNPs to facilitate efficient homologous integration at the target loci To prevent the potential selection of ectopically integrated functional copies of the genes primers were designed to omit either the 5′ or 3′ or both ends of the coding sequences for particularly short genes (BcoliC and Bcmix17) full length products of the coding sequences were transformed because of the requirement of having sufficiently long flanking regions Complete amplicons of coding sequences were also used if previous transformation attempts with truncated sequences did not yield resistant colonies To prevent the identification of false positive mutations wild type (B05.10) amplicons of the respective sequences were transformed as negative controls Bcpos5 and Bcmdl1 alleles from AniR1 field samples were amplified from the following sources: Bcpos5V273I: 10-Bc-187 In order to narrow the AniR1 mapping window of cross 09Bc11 x B05.10 (see previous section Mapping of Resistance Loci in Field Isolates and Phenotypic Characterization) and identify the genetic factor conferring the resistance phenotype 10 overlapping fragments of ~11–12 kb length from genomic DNA of the resistant parent 09Bc11 and of the sensitive parent B05.10 as negative control were amplified by PCR and transformed into B05.10 as described below (primers listed in Table S3) For this approach LongAmp Taq DNA Polymerase (NEB Based on the sole fragment amplified from 09Bc11 that conferred resistance the same strategy was reiterated using smaller overlapping fragments of ~3.6 kb (Table S3) Prior to transformation PCR products were purified using the NucleoSpin® Gel and PCR Clean-up kit (Macherey-Nagel), using between 2 and 8 pmoles (6–16 μg) of DNA for transformation. The method for PEG-based transformation of B. cinerea protoplasts was adapted from a protocol developed for Fusarium oxysporum (Malardier et al., 1989) NY medium in round-bottom flasks was inoculated with 108 conidia of B05.10 per 100 ml and shaken for 16 h at 180 rpm The cultures were centrifuged for 10 min at 3,700 g and the germlings were washed twice in KCl/NaP buffer (545 mM KCl 3.5–4 g germlings (wet weight) were re-suspended in 40 ml KCl/NaP buffer and 0.5 g Lysing Enzymes from Trichoderma harzianum (Sigma After 2.5 h protoplasts were filtered through 25 μm nylon meshes and washed in ice-cold TMS buffer (1 M sorbitol; 10 mM MOPS Protoplasts were then re-suspended in ice-cold TMSC buffer (TMS plus 40 mM CaCl2) and the concentration (hemocytometer) adjusted to 2·107 protoplasts per 100 μL The DNA to be transformed was prepared in 100 μL aliquots in TE buffer supplemented with 40 mM CaCl2 and mixed with 100 μL protoplast suspension 160 μL of PEG6000/MS solution (1.2 g PEG6000 in 800 μL MS: 0.6 M sorbitol; 10 mM MOPS pH 6.3) was added and carefully mixed with the protoplasts After another 15 min incubation at room temperature 700 μL TMSC buffer was added and the mixture was centrifuged for 5 min at 1,000 g The protoplasts were re-suspended in 200 μL TMSC and added to 200 mL SHA (42°C) containing CDL (10 mg·L−1) and 20 mL aliquots poured into Petri dishes The plates were incubated at 20°C in the dark for up to 3 weeks and examined for the emergence of resistant colonies which were picked and further propagated on Vogel's minimal agar with CDL (10 mg·L−1) Homologous recombination of the transformed truncated or full-length gene copies within putative transformants was subsequently tested by amplification of the genomic regions encompassing the homologous recombination points (for PCR primers see Table S3) An accurate determination of EC50 values for the latter two types of curves was not possible Types of dose-response curves of CDL-resistant UV mutants observed in liquid culture tests The graphs show mean values of triplicate measurements ± standard deviation (A) Classical monophasic dose-response curves of a CDL sensitive strain (squares) and a resistant UV isolate (circles) (B) Very slightly biphasic curve (based on visual ranking) (C) Biphasic dose-response (no regression curve) (D) “Displaced” biphasic dose-response (no regression curve) A collection of 13 UV mutants displaying stable resistance and covering each of the four distinct dose-response phenotypes together with the non-mutagenized reference isolate Comparison of the sequencing data with the annotated B05.10 reference genome (see Materials and Methods) revealed single nucleotide polymorphisms (SNPs) and short deletions in the genomes of the mutants that were not found in the non-mutagenized control By filtering the results at high stringency up to five non-synonymous SNPs within different predicted genes were identified per UV isolate (Table S6) No such mutations were found in strain CDL10-11 although displaying a stable loss of CDL sensitivity In an attempt to reveal non-synonymous variations occurring at a lower frequency further analysis of the sequencing data of CDL10-11 was undertaken This led to the identification of premature stop codons within multiple genes including Bcin03g01010 which encodes the mitochondrial inner membrane ADP/ATP carrier The SNP within this gene was only detected in 46% of the sequencing reads (data not shown) were found to confer resistance even when the transformed PCR products contained coding sequences truncated at either the 5′ or 3′ ends to force homologous integration events The other four required full length gene copies either because of their small size (Bcmix17 and BcoliC) or because previous transformation attempts with truncated gene fragments failed for unknown reasons (Bcatm1 and Bcmcr1) This full length gene approach presents the disadvantage of possible ectopic integrations of additional functional copies into the genome (not tested) The mutation detected at low frequency within strain CDL10-11 resulting in a premature stop codon within Bcin03g01010 was tested in the same way but only as a 5′-truncated gene copy which did not result in resistant transformants Validated CDL resistance-conferring mutations and sensitivity data of UV mutants they ranged between 142 and 269-fold for Bcmdl1E407K and Bcafg3L305P This may be caused by the presence of a predicted 35 kb centromeric region (1,132,000–1,167,000) at the center of the window PCR products used to identify the R locus by a reverse genetic approach are represented by black or red bars where red bars indicate fragments conferring R when transformed into the S strain B05.10 (D) Reverse genetic validation transformation with genomic fragments Bcin10-1-1 and Bcin10-1-2 (amplified from 09Bc11) transformed into B05.10 and selection on CDL-containing agar The latter fragment contains the resistance-conferring mutation in gene Bcin10g02880 An iterative reverse genetic procedure was used to further delimit the genetic factor conferring the AniR1 phenotype (Figures 2B–D, see Materials and Methods, and Table S3). The smallest fragment, amplified from the resistant strain 09Bc11, that resulted in resistant colonies after transformation into the sensitive reference strain was Bcin10-1-2 (1,077,456–1,081,069; Figures 2C,D) The sole non-synonymous SNP within this region is located at position 1,078,999 on chromosome 10 and leads to an L412F substitution within the protein encoded by the gene Bcin10g02880 (termed Bcpos5 hereafter) The encoded BcPos5 protein is the Botrytis ortholog of Pos5 more than one third of the AniR1 samples (35%) remained unexplained (A,B) Partition of Bcpos5L412F and Bcmdl1E407K as inferred from pyrosequencing within 491 AniR1 samples collected between 2009 and 2014 from grape and strawberry (C) Box-plot representation of the distribution of CDL sensitivity for the Bcpos5 and Bcmdl1 genotypes within the monitoring population isolated in 2014 The different genotypes were either detected by pyrosequencing (G408R/V (D) Effect of MDR on the CDL sensitivity range for WT and Bcpos5L412F genotypes based on the 2014 population (Table S1) Results of unpaired t-tests (GraphPad Prism 6.0) comparing Bcpos5WT with or without MDR and Bcpos5L412F with or without MDR (*p < 0.0001 Validated and proposed CDL resistance-related mutations identified in field samples of B Despite AniR1 phenotypes could be explained for all samples collected from grape and strawberry in 2014 the characterization of confirmed AniR1 samples from tomato suggests that additional resistance mechanisms outside Bcmdl1 and MDR may occur in the field at a low frequency (Table S2) For reasons that are not understood the transformation of truncated versions of the Bcpos5 gene followed by direct selection on CDL-amended plates failed L412V being the only allele giving satisfactory results with this approach Sanger sequencing of individual transformants showed that the L412V mutation successfully replaced the wild type allele at the Bcpos5 locus the wild type allele remained present for all other Bcpos5 mutant alleles tested suggesting that resistance was conferred by the ectopic integration of functional copies at unknown genomic loci in these transformants both E407K and S466R resistance-conferring mutations of Bcmdl1 successfully replaced the wild type copy indicating correct homologous recombination events in the transformants Validation of CDL resistance-conferring field and UV mutant genotypes in Bcpos5 and Bcmdl1 by reverse genetics Result of a transformation of reference strain B05.10 protoplasts with 8 pmoles of PCR products amplified from genomic DNA of field isolates (UV mutant CDL50-8 in case of Bcmdl1G422R) carrying putatively resistance-conferring SNPs on selective plates supplemented with CDL (10 mg·L−1) (A,B) Bcpos5 PCR products transformed included the complete coding sequences Pictures taken 12 or 19 days after transformation truncated at 5′ and 3′ ends Bcmdl1E407K conferred slightly but significantly higher resistance levels compared to Bcpos5L412F (p < 0.0001) but the combined Bcpos5L412F + Bcmdl1E407K resistance levels were similar to those displayed by Bcmdl1E407K alone (p = 0.99) Phenotypes of progeny isolates from a cross between Bcmdl1E407K and Bcpos5L412F Box and whisker plots show EC50 values measured in technical duplicates (at BIOtransfer) Results of unpaired t-tests (GraphPad Prism 6.0) between two of the progeny genotypes are indicated (*p < 0.0001 but given the importance of genetic background additional homologous recombinants for Bcpos5G408R/V and Bspos5L412F will be required to validate these potential variations in cross resistance profiles Cross-resistance profiles of a collection of isolates including the most prevalent AP resistance mutations from the field and available corresponding homologous recombinants Resistance factors were calculated relative to the reference strain B05.10 Sensitivity to FDL was tested to confirm the absence of MDR1/3 (multidrug resistance) phenotypes (A) Individual single spore isolates with Bcpos5 and Bcmdl1 genotypes confirmed by Sanger sequencing Mean values of technical triplicates ± standard deviation of one isolate per mutation are shown mean values of three transformants per genotype ± standard deviation are shown Individual transformants were tested in technical triplicates only a proportion of the sequencing reads showed non-synonymous mutations within multiple genes was found in the ortholog of the gene encoding the yeast mitochondrial inner membrane ADP/ATP carrier Pet9 Considering the mitochondrial localization of the validated resistance-related genes this likely loss of function mutation may be responsible for CDL resistance in this isolate but a reverse genetic experiment failed to confirm this hypothesis which both affect the C-terminal part of the protein and lead to polypeptide sequences that are 15 amino acids longer (Figure S1) These biphasic behaviors suggest a complex interplay between the primary mode of action and the likely suppressive/compensatory mechanisms caused by the identified mutations in these genes We assume that dose-response curves of the mutants may be indicative of the functional distance between the mutated gene product and the primary target We would expect from target binding site mutants to directly suppress growth inhibition at low doses of the fungicide biphasic dose-responses show that a primary inhibitory effect at low fungicide concentrations is not prevented This is suggesting that in such cases resistance is conferred by a suppressive effect or compensatory mechanism functionally related to the molecular target candidates for the primary target would include Bcpos5 and Bcdnm1 both of which displayed no sign of biphasicity in our assays Bcpos5 was found to explain most of the AniR1 resistance in the field populations but was not identified in our UV mutant collection resistance through mutated versions of this gene should have been found in our UV screening a wide range of Bcpos5 mutations were shown to confer resistance in reverse genetic experiments but homologous recombination was only observed for one allele (L412V) whereas ectopic integrations were obtained for all other alleles tested These results might indicate that the resistance phenotype caused by the majority of Bcpos5 mutations would have required more time to be fully expressed than was allowed by the direct selection of UV mutants and transformants on agar plates supplemented with CDL this hypothesis will require further validation It is also possible that mutants in Bcpos5 occurred at a low frequency in our UV screening because not all isolates were genetically characterized All 9 CDL resistance-related genes identified in this study are nuclearly-encoding proteins targeted to the mitochondria and involved in various mitochondrial functions (Figure 7, Table 3) suggesting that the mitochondrion is the primary target of this class of fungicides some of these proteins have been functionally connected and shown to either physically or genetically interact in yeast and other systems the oligomycin resistance-conferring subunit c of the F1FO ATP synthase and Mdl1 are involved in mitochondrial respiration and energy production through their role in the folding maturation and degradation of internal mitochondrial membrane (IMM) proteins and in the transport of peptidic degradation products by Mdl1 from the mitochondrial matrix to the intermembrane space (IMS) Model of hypothetical interactions between the CDL resistance-related mitochondrial proteins Mitochondrial unfolded protein response; EAFR Positions of resistance-related mutations are marked with red stars Homologous gene products of CDL resistance-related B cinerea genes and their reported cellular functions The mutation may therefore functionally mimic BcPhb2 mutations if the BcPhb2-BcAfg3 interaction effectively accounts for resistance It is tempting to speculate that the R33C mutation of the BcOliC targeting peptide leads to AP resistance by interfering with an additional biological function carried by the targeting peptide itself Haf1 was shown to be essential for channeling peptides derived from degraded mitochondrial proteins from the mitochondria to the cytosol and leading to the re-localization of a bZIP protein (ZC376.7) from the cytosol to the nucleus Although such mitochondrial to nuclear peptide signaling has never been demonstrated in B cinerea one can hypothesize a role of the resistance-conferring mutations of BcMdl1 in interfering with the transport of particular peptides or signaling molecules One can also hypothesize that the BcMdl1 transporter specificity is modified in a way that effectively supports the transport of AP molecules out of the mitochondrial matrix in resistant BcMdl1 mutants this would be the first example of mitochondrial multidrug resistance mutated at positions F412 and G408 in BcPos5 correspond to positions L397 and G393 of yeast Pos5 These positions are located within a short distance (8 and 9 Å respectively) from the NAD+ ribose phosphorylation site suggesting that these mutations could have an influence on catalysis the Pos5 ATP binding site has not yet been described and these residues may also be involved in the binding interaction with ATP and P319A of BcPos5 correspond to positions I243 They are located at more distant sites and could possibly interfere with the formation of the tetramer or its potential allosteric regulation Given the degree of connectivity of Pos5 with multiple other biochemical pathways and the enzymatic substrates or products of many of the resistance genes we identified it is tempting to hypothesize that this enzyme is the primary target for AP fungicides Further work focusing on this aspect is ongoing additional AP resistance-conferring mutations identified in our UV screening were located within Bcmix17 the function of these genes appears less clearly connected to the core pathways described above We hypothesize a possible link between these mutations and the sensing of substrates transported by BcMdl1 or BcAtm1 Our AP-resistant BcMcr1 mutants were frameshift variants of the C-terminal part of the protein which was extended by 15 residues for both mutants identified These frameshifts are unlikely to influence the dual sub-organellar localization of the protein but might impact enzyme kinetics or the nature of the reduced substrates which could be different depending on the sub-organellar localization The very unique growth inhibition dose-response curves displayed by these variants (Figure S1) suggest a complex interplay between the mode of action of APs and the concentration of substrates or products of this enzyme We report resistance toward one single class of fungicides caused by mutations within at least 9 different genes suggesting novel functional connections that are likely to be conserved across species and therefore extremely relevant for cell biology Such a diversity of possible resistance mechanisms for a single fungicide class is and despite the fact that the majority of AniR1 field resistances could be explained it is clear that we did not identify all possible resistance mechanisms It is likely that other mutations within other genes would constitute a good candidate for being the primary molecular target of AP fungicides the very similar cross-resistance profiles across a panel of Bcpos5 mutants did not suggest strong differential binding interaction across the different fungicides and mutant pairs which is in accordance with the very similar structure carried by APs further work will be necessary to confirm the molecular target and the exact mechanism of action of this important class of fungicides a better knowledge of the relationships between the molecular target and the multiple resistance mechanisms will be required to understand whether or not the various resistance mutations represent a gain or a loss of function within the identified resistance genes the identification of AP resistance-conferring mechanisms within field populations of B cinerea will facilitate the development of quantitative molecular assays for resistance monitoring This new knowledge will also enable the assessment of the stability and fitness of the different resistant variants of Bcpos5 and Bcmdl1 Since resistance toward AP fungicides has also been described in other species and is in many cases infrequent within populations it will be interesting to determine whether similar genes are affected or not and whether such resistance mechanisms encompass species-specific fitness costs GS and AM conceived the project and designed the experiments and SW designed and performed next generation sequencings and did the bioinformatic analysis TB designed and supervised the Botrytis monitoring (phenotyping assays) at BIOtransfer 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 and Helge Sierotzki for reviewing the manuscript the Netherlands) for his support with genetic crossings of B Italy) for kindly providing strain BAR633 and Eddy Blondet (BIOtransfer) for conducting the Botrytis monitoring assays The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fmicb.2017.02361/full#supplementary-material 1. ^Fungicide Resistance Action Committee (FRAC): http://www.frac.info/ Structural determinants of discrimination of NAD+ from NADH in yeast mitochondrial NADH kinase Pos5 The formation of respiratory chain complexes in mitochondria is under the proteolytic control of the m-AAA protease an AAA protease with chaperone-like activity in the inner membrane of mitochondria Evidence for a novel mitochondria-to-nucleus signalling pathway in respiring cells lacking i-AAA protease and the ABC-transporter Mdl1 A mutation in the mitochondrial fission gene Dnm1l leads to cardiomyopathy 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mitochondrial protein import Google Scholar French vineyards provide information that opens ways for effective resistance management of Botrytis cinerea (grey mould) The ATP synthase subunit 9 gene of Aspergillus nidulans: sequence and transcription Role of the ABC transporter Mdl1 in peptide export from mitochondria Corran A and Scalliet G (2017) Anilinopyrimidine Resistance in Botrytis cinerea Is Linked to Mitochondrial Function Received: 21 August 2017; Accepted: 15 November 2017; Published: 30 November 2017 Copyright © 2017 Mosbach, Edel, Farmer, Widdison, Barchietto, Dietrich, Corran and Scalliet. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY) *Correspondence: Andreas Mosbach, YW5kcmVhcy5tb3NiYWNoQHN5bmdlbnRhLmNvbQ== Gabriel Scalliet, Z2FicmllbC5zY2FsbGlldEBzeW5nZW50YS5jb20= Disclaimer: 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 94% of researchers rate our articles as excellent or goodLearn more about the work of our research integrity team to safeguard the quality of each article we publish Piatt County Miss Contestants 2015 – Sydney Mumm Piatt County Parade of Princess contestants 2015 – Kali Griffith The Piatt County Fair Queen pageant will be held at 7 p.m June 16 at the Piatt County Fairgrounds in Cerro Gordo Parade of Princess contestants – Kali Griffith 2014 Junior Miss Piatt County; Riley Brandenburg Your browser is out of date and potentially vulnerable to security risks.We recommend switching to one of the following browsers: The Naples Holiday Shootout is returning to its original home The annual holiday girls basketball tournament bringing elite teams and talent to Southwest Florida is back at Barron Collier but has changed venues three times since then — spending three years at St one at Community School and the last two at Gulf Coast Tournament director Ron Mosbach said the reason for this year’s switch is pretty simple — it’s much easier logistically for him Mosbach is the freshman/JV and assistant varsity coach at Barron Collier “We’re really excited to have it back here,” Mosbach said “Back where it started and back where we feel it should be Gulf Coast was a great tournament host and gave us everything we needed, but with their team graduating all those seniors and wanting to play in a tournament away from the area although the event will take just three days to complete Since a few of the schools don’t have permission to play on Sundays tournament organizers are working with FGCU to send the tournament teams to the Duke-FGCU game that afternoon Although the tournament has switched venues frequently in recent years Defending champion Miami Country Day is back coming off a season in which the Spartans went 26-2 and finished the year ranked No Also in the field is Georgia Class 7A champion Atlanta-Westlake Six defending state champions will be competing and most of the 16-team field made deep postseason runs a year ago Host Barron Collier and perennial state power Fort Myers are the only two local squads in the field “It’s a great event that brings top talent to Naples,” Mosbach said “And we usually have at least one team that seemingly brings its entire community with it it’s an opportunity to go against a level of competition we wouldn’t see otherwise.” Marco boys building cross country powerhouseMarco Island Academy is a small school but it's becoming quite big on the local cross country landscape The Manta Rays won their second district championship in three years claiming the Class 1A-District 6 title at Southwest Florida Christian Academy on Thursday The Manta Ray boys own the only team titles in any sport for Marco Island Academy "This definitely helps build momentum for the athletic program moving forward," Marco Island athletic director Ryan Marie Roberts said "For a tiny school like us to be able to put a championship-caliber team together is definitely an incredible accomplishment." Now the Manta Rays have their sights set on becoming the first team in school history to qualify for the state championships in any sport Marco Island competes in the regional meet Friday in Lakeland we missed it by one spot," Marco Island coach Bruce Kretschmer said "It'll all be down to keeping everybody healthy As long as we don't have any major injuries or illnesses Definitely the most competitive group I've ever coached." The Manta Rays got a tremendous grouping at the district meet with junior Kevin Barry leading the way by finishing eighth Junior Johnathan Witt was right behind him in ninth Rounding out the Manta Rays' top five were sophomore Tony Castillo (18th) and senior Damien Tschida (21st) "We figured Southwest Florida Christian would be our top competition and when I saw their top two kids came in second and third overall fourth and fifth kids really came through for us Junior Skylar English announced her verbal commitment to FGCU via an Instagram post Saturday English is among the Cougars' leaders in every statistical category topping the squad in kills (463) and aces (65) She's second on the team in blocks (37) and third in digs (232) has been an integral contributor to a pair of state semifinalists for Barron Collier The Cougars advanced to the championship match two years ago and lost in the semifinals last year Barron Collier (24-5) is back in the regional championship this year hosting Archbishop McCarthy on Tuesday.  Naples running back Elan Sommala was selected as the Class 6A FACA District 18 Player of the Year highlighting the list of Collier County players honored Sommala has amassed 1,253 yards on 182 carries with 14 touchdowns in nine games Offensive lineman Mike Fangman and tight end Dominic Mammarelli were the other first-team offensive selections for Naples linebacker Ar'Quel Smith and defensive back Brady Webre were first-team defensive selections Coach Bill Kramer was selected the Class 6A coach of the year and Gulf Coast's Tom Scalise was selected as the Class 7A coach of the year The insolvency proceedings over Bama GmbH have been lifted The district court in Mosbach confirmed this on July 31st The Mosbach-based furniture provider had previously found a new investor in the Hamburg-based MKCP Beteiligungsgesellschaft mbH “The takeover of Bama GmbH by MKCP is an important milestone in continuing our business activities and fully concentrating our energies on operational business,” the company said “The successful completion of the procedure marks an important step in the history of our traditional company,” commented Managing Director Dr Bama filed an application for self-administration insolvency proceedings last November The reason for the application was “significant market distortions and the resulting decline in sales,” it was said at the time We always keep you up to date: with our free newsletter SHOEZ compact You will regularly receive all information from the shoe industry in a clear form when a new magazine is published US Managing Director Peter Sachs hands over to Lance Taylor Alchemy plans to take over almost half of the Austrian shoe retailer Second best financial year in the company's history Creditors' meeting decides against P&C's takeover offer Sanela Krisat becomes International Sales Director sales representatives and business partners throughout the DACH association Telephone: +49 (0) 6 41 / 7 95 08 – 0Fax: +49 (0) 6 41 / 7 95 08 – 15Email: info@shoez.biz Designed by willsch-media.de franceimage © hisao suzukiall images courtesy of louvre-lens  positioned approximately 200 kilometers north of paris, the center intends to bring touristic attention to a collection of museums within the industrial region of nord-pas de calais. maintaining the openness of the once mine works site the 28,000 square meter building has been broken into smaller spaces to follow the gradual grade changes of the terrain glass-enclosed galleries open perspectives across the grounds using transparency to create cross views through public spaces spanning 306 meters in length and following the long and curved shape of the property, a subtle distortion is generated inside without disrupting the display of artwork polished aluminum reflects a blurred image of the undulating contours full-height bays of glass bring in daylight while a system of roll down shades protect the artwork supporting the metal roof structure are many slim steel columns painted white to abandon the practice of departmentalization a 3,000 square meter gallery called the ‘grande galerie’ will exhibit artifacts from the louvre’s collection without partitions along its 120 meter length a 1,000 square meter glass pavilion will be a relaxing atmosphere to experience themed exhibitionswhich examine the concept of time the extensive park overtakes the 20 hectares of coal-mining landscape with nature bringing a role of sustainability into the palette of artistic facilities to support the daily and logistical functions see designboom’s original coverage of this project during schematic phase.  image © iwan baan landscape planimage © catherine mosbach floor plan / galerie du tempsimage © SANAA section / galerie du tempsimage © SANAA sectionimage SANAA © SANAA / imrey culbert / catherine mosbach schematic renderingsimage © SANAA / imrey culbert / catherine mosbach image © SANAA / imrey culbert / catherine mosbach interior perspectiveimage © SANAA / imrey culbert / catherine mosbach exhibition roomimage © studio adrien gardere AXOR presents three bathroom concepts that are not merely places of function but destinations in themselves — sanctuaries of style Catherine Mosbach is the founder of Paris-based design firm mosbach paysagiste Catherine graduated from the École Nationale Supérieure de Paysage of Versailles in 1986 after studying biological sciences and physical chemistry at the University Louis Pasteur Strasbourg She is renowned for socially and environmentally responsible work that attests to temporality and continuing change referring those who interact with these landscapes to relationships with history She was a speaker at this year’s Living Cities Forum places and the people who make them delivered direct to your inbox A Naples man was arrested early Sunday morning after crashing during a high-speed chase on Vanderbilt Beach Road was spotted by Collier County sheriff's deputies street racing in a stolen 2016 GMC Yukon on Immokalee Road near Airport-Pulling Road at 4:20 a.m said sheriff's spokeswoman Jamie Mosbach. The SUV was reported stolen from Windstar on Naples Bay The vehicle sped toward the beach chased by deputies before smashing into concrete benches on Gulf Shore Drive Mosbach said.The pursuit reached speeds in excess of 100 mph Lopez was later found and arrested by deputies near the beach driving while license is suspended or revoked and resisting arrest without violence He was taken to NCH North Naples Hospital for evaluation, Mosbach said.  Even though Lopez faces a street-racing charge deputies Sunday said they had no information on whether another vehicle was involved You are using an outdated browser. Please upgrade your browser to improve your experience the 25-member University of Guelph Chamber Singers and conductor Marta McCarthy travelled to Mosbach to compete in the Mosbach International Competition for Chamber Choirs the Guelph choir was chosen to be one of three finalists who would perform a second time Frieder Bernius and Marcus Creed – who are world-renowned – praised choir members for their passion and their unique tone quality,” says McCarthy “It was a tremendous accomplishment to win third prize especially since the other two choirs in the finals were the premiere choirs of universities devoted entirely to music: Hochschule fur Musik von Detmold and Hochschule fur Musik von Mannheim All the choirs were of extremely high calibre.” the experience of performing in the competition was the most important aspect; placing third really felt like icing on the cake “We have never sung better or with as much passion reflectiveness and enthusiasm as we did during the competition It was wonderful to meet and learn about different cultures from the many members of the choirs and it was an incredible learning experience filled with new sounds Choir member Jerome Chang describes the trip as “an opportunity of a lifetime One of the most memorable evenings was after the competition and gala when all the choirs from around the world came together to have an informal singalong We couldn’t all speak each others’ languages but we could definitely share music together – over good food and drink.” Craig Bechtel also has great memories of singing with the other choirs “It was an amazing experience; everyone there had a positive and professional outlook on the competition.” Another member of the choir describes feeling proud to be part of this hard-working and talented group of people The choir members are also appreciative of the generosity of others who made the trip possible “I appreciate that the dean and the University gave us financial assistance to bring this trip to fruition It was wonderful being an ambassador of the University of Guelph in Europe and I know the University would have been proud of us.” The Mosbach competition began with each choir performing the first of two 25-minute programs of highly challenging repertoire The top three choirs performed a second program they also performed for appreciative audiences at a church service in Mosbach and at a gala concert celebrating the music of each country represented in the competition The Canadian repertoire performed by the University of Guelph choir was especially warmly received She adds: “The atmosphere at the competition was extremely positive with each group cheering for the others and truly using the experience to learn and grow which included songs from each culture and joyful intermingling between the choirs We were proud to be musical and cultural ambassadors for Canada.” “Your marvellous choir sound!” That’s how the University of Guelph Chamber Singers impressed the audition jury of the Mosbach International Chamber Choir Competition This success led to an invitation to fly to Germany this April as one of only six choirs chosen – and the only one from North America The judges for this event will be the world-renowned conductors Frieder Bernius and Marcus Creed For the 26 singers in the choir and conductor Marta McCarthy this is an exciting opportunity to perform in an exclusive setting and broaden their cultural horizons simply means a small choir of less than 40 members The U of G chamber singers include 14 men and 13 women; the longest-standing participant is bachelor of arts and science program advisor Jerome Chang and the two newest are undergrad Tyler Myles and post-doc Ben Wright the chamber singers are committed to contributing to their community in meaningful ways In addition to their regular concert series each year they participate in fundraising concerts for charities such as the United Way The U of G singers were recommended to Mosbach organizer Christof Roos by Leonard Ratzlaff who was one of the judges when the Guelph singers won the collegiate category of the 2011 National Radio Competition for Amateur Choirs McCarthy secured the invitation by helping the choir prepare an audition CD that met the requirements for three contrasting pieces recorded within the last two years Roos told McCarthy: “Your choir will compete very well among the other choirs We are very proud to be presenting a choir coming from so far to our public The other choirs will all come from Europe.” being accepted into a competition like this is just the first step McCarthy had to develop a repertoire of songs that followed the guidelines Each choir has to prepare both a first-round program and a second-round program but only three of the six will go on to the second round “There is a compulsory song for each round that all the choirs have to do,” says McCarthy “That gives them a standard to compare against.” Choirs must also do a song in each program that dates back to before the 1800s McCarthy has added a Canadian song to each of her programs The Canadian pieces will be The Solo by Ruth Watson Henderson and O Magnum Mysterium by Jeff Enns The choir is delighted to represent U of G in this competition McCarthy says she and the students are especially grateful for the financial support of the College of Arts dean’s office which underscores the value of this exceptional learning experience “We will be able to hear other fine choirs and hear many songs that will be new to us will probably have a very different sound,” McCarthy says “The opportunity to see people performing at a very high level is always valuable.” If you’d like to have a sneak preview of their German performance, the choir is performing at Harcourt Memorial United Church on March 31 at 8 p.m. Tickets are $10 and will help to fund the costs of travelling to Germany. The women’s choir and the symphonic choir will also be performing. Email Vicki Isotamm at visotamm@uoguelph.ca for tickets Click below to hear them sing O Magnum Mysterium The experts were unanimous - digital transformation is needed now This was also evident at the panel discussion "Digitalization concerns everyone" which MPDV hosted at the Baden-Wuerttemberg Cooperative State University (DHBW) Mosbach to mark World Smart Factory Day in September  "Technology alone is not enough - we must be prepared to accept and use it," emphasized Nina Warken Member of the Bundestag and Parliamentary Secretary of the CDU/CSU parliamentary group in her opening statement at the panel discussion "The innovative strength of SMEs will be decisive in overcoming competitive pressure," added Josef Stumpf Director of the Federal Economic Senate for the Baden-Württemberg region Managing Director of Perfect Production GmbH referred to the enormous advantages of networked data in companies and the increasing importance of professions such as data analysts who integrate artificial intelligence and automation into operational practice Managing Director of the Rhine-Neckar Chamber of Industry and Commerce also highlighted the potential of digitalization to reduce bureaucracy and make processes more efficient Under the motto "Digitalization concerns everyone" the well-attended event in the DHBW's Audimax offered an examination of the future of digitalization The focus was on the question of how Germany is positioned internationally in Industry 4.0 and what skills are required for a successful transformation who engaged in an intensive exchange about the Internet of Things (IoT) and the transformation of industry The panel included the aforementioned representatives from various areas of society as well as Prof Dean of the Faculty of Engineering at DHBW Mosbach The panel discussion was moderated by Alexandra Gorsche who ensured a lively debate by asking specific questions opened the event with an inspiring speech about the importance of practical education for Industry 4.0 a long-standing partner company that had already made the "Living Lab" at the DHBW possible in 2015 - a model factory equipped with modern IoT technologies that was named one of "100 Places for Industry 4.0" aims to inform as many people as possible about the importance and possibilities of digitalization and the Smart Factory in particular emphasized the company's social responsibility: "Our goal is to inspire not only companies but above all the next generation for the opportunities of the smart factory Digitalization and sustainability go hand in hand and offer a unique opportunity to make our future worth living." Source and further information: www.mpdv.com 15 Dec 2022 • 2m read • View Author Public-park-proposals-sought-for-Barangaroo-1732008451.png Infrastructure NSW is calling for registrations of interest from designers interested in designing Harbour Park the park’s site sits on the Sydney Harbour foreshore says the park is the next chapter in the evolution of Barangaroo we’re looking for ideas that embrace Barangaroo’s commitment to design excellence and sustainability while still ensuring the park celebrates the natural history and living culture of the site,” he says Former Prime Minister Paul Keating will Chair the Jury for the competition which features Australian landscape architect and urban designer Oi Choong Greenaway Architects’ Founder Jefa Greenaway along with French landscape architect from mosbach paysagistes Choong says the opportunity presented to landscape architects and urban designers is equally rare and exciting you have a blank canvas for greening the heart of Barangaroo adding to the necklace of parklands and icons that define our world-famous harbour,” she says to lead the design of this significant public space at Sydney’s latest landmark waterfront destination - it is a project that will be a grand contribution to Sydney following on the success of Barangaroo Reserve.” Entrants must consist of Australian-led design teams with international and interdisciplinary talent within the teams permitted Competitors will first be shortlisted based on their credentials and general approach before embarking on a design concept that completes the vision for Barangaroo’s open public spaces and continues the precinct’s commitment to honouring the role of First Nations people and the history of the site “We’ll be seeking innovative ideas for a flexible and active space that will add to the art and cultural offerings already in Barangaroo Registrations of interest into the Harbour Park design competition are now open. Those wanting to express their interest can do so here. BOSS Pedestal Planner: Take the guesswork out of pedestal estimation Anston’s brand rejuvenation consolidates architectural products, streamlines operations Case study: Rifle Range Retrofit, Melbourne, VIC EPDs in construction: Building transparency, trust, and sustainability Sign up to our newsletter for the latest industry news, products and inspiration.