The conserved XPF:ERCC1-like Zip2:Spo16 complex controls meiotic crossover formation through structure-specific DNA binding.

In eukaryotic meiosis, generation of haploid gametes depends on the formation of inter-homolog crossovers, which enable the pairing, physical linkage, and eventual segregation of homologs in the meiosis I division. A class of conserved meiosis-specific proteins, collectively termed ZMMs, are required for formation and spatial control of crossovers throughout eukaryotes. Here, we show that three Saccharomyces cerevisiae ZMM proteins-Zip2, Zip4 and Spo16-interact with one another and form a DNA-binding complex critical for crossover formation and control. We determined the crystal structure of a Zip2:Spo16 subcomplex, revealing a heterodimer structurally related to the XPF:ERCC1 endonuclease complex. Zip2:Spo16 lacks an endonuclease active site, but binds specific DNA structures found in early meiotic recombination intermediates. Mutations in multiple DNA-binding surfaces on Zip2:Spo16 severely compromise DNA binding, supporting a model in which the complex's central and HhH domains cooperate to bind DNA. Overall, our data support a model in which the Zip2:Zip4:Spo16 complex binds and stabilizes early meiotic recombination intermediates, then coordinates additional factors to promote crossover formation and license downstream events including synaptonemal complex assembly

Submarine groundwater discharge: an unseen yet potentially important coastal phenomenon

In collaboration with researchers from Florida State University, Florida Sea Grant introduces an important but poorly known topic: submarine groundwater discharge. Although nearly invisible, submarine groundwater discharge influences coastal systems. This brochure helps explain this important phenomenon. (8pp.

Conformational dynamics of the Hop1 HORMA domain reveal a common mechanism with the spindle checkpoint protein Mad2.

The HORMA domain is a highly conserved protein-protein interaction module found in eukaryotic signaling proteins including the spindle assembly checkpoint protein Mad2 and the meiotic HORMAD proteins. HORMA domain proteins interact with short 'closure motifs' in partner proteins by wrapping their C-terminal 'safety belt' region entirely around these motifs, forming topologically-closed complexes. Closure motif binding and release requires large-scale conformational changes in the HORMA domain, but such changes have only been observed in Mad2. Here, we show that Saccharomyces cerevisiae Hop1, a master regulator of meiotic recombination, possesses conformational dynamics similar to Mad2. We identify closure motifs in the Hop1 binding partner Red1 and in Hop1 itself, revealing that HORMA domain-closure motif interactions underlie both Hop1's initial recruitment to the chromosome axis and its self-assembly on the axis. We further show that Hop1 adopts two distinct folded states in solution, one corresponding to the previously-observed 'closed' conformation, and a second more extended state in which the safety belt region has disengaged from the HORMA domain core. These data reveal strong mechanistic similarities between meiotic HORMADs and Mad2, and provide a mechanistic basis for understanding both meiotic chromosome axis assembly and its remodeling by the AAA+ ATPase Pch2/TRIP13

Revising old child support orders: The Wisconsin experience

In an effort to make Wisconsin's child support cases more equitable and up-to-date, child support staff reviewed "old" child support orders in thirteen of the state's seventy-two counties. (Reviewing old child support orders is now mandatory under the provisions of the Family Support Act of 1988.) Of the reviewed cases, only 21 percent were revised. Primary reasons for non-revision were the economic circumstances of the noncustodial parent (among welfare cases) and a lack of permission by the custodial parent to proceed (among non-welfare cases). Revised orders increased substantially, an average of $116/month (77 percent). An alternative method of keeping orders current is to express them as a percentage of the noncustodial parent's income; these orders are kept up-to-date automatically and are associated with large increases in collections.

Adolescent and Adult Perceptions of Adolescent Diet, Physical Activity, Body Size, and Obesity Prevention in Botswana

Obesity is a worldwide health problem affecting developed and developing nations. Botswana is an upper-middle income nation experiencing rapid urbanization, economic, and nutritional change. The combined prevalence of adolescent overweight and obesity has now surpassed underweight. Little is known regarding the adolescent and adult perceptions of adolescent diet, physical activity, body size, the meaning of obesity, and obesity prevention interventions. This qualitative-descriptive study uses Ecological Systems Theory to analyze and interpret data from 15 focus groups (12 adolescent and 3 parent focus groups of unrelated participants) conducted in Gaborone, Botswana. The purpose of this study is to describe the factors that influence adolescent and adult perceptions and attitudes related to adolescent diet, physical activity, body size, obesity and potential obesity interventions. Prolonged data immersion, clustering of analogous data into themes, and direct reporting of participant voices were used to provide a rich description of the shared experiences and perceptions of the study participants. Results indicate that adolescents identify healthy versus unhealthy foods, yet choose unhealthy foods based on taste preferences, social pressures, and the perceived affordability of unhealthy foods. Parents prefer a healthy diet for their adolescent children, although they acknowledge purchasing unhealthy foods for their children based on adolescent taste preferences and social pressures. Adolescent and parent participants suggest a body size analogous to normal weight is most desirable for adolescents themselves and most attractive in the opposite sex. Obese body sizes are associated with perceived laziness, isolation, and negative social stigma. Additionally westernized preferences are replacing the historical notion that a large body size is consistent with wealth, strength, and prosperity. Participants also perceive thinness to be related to illness, specifically suspected HIV infection due to the relatively high prevalence of HIV infection in Botswana. Adolescents and parents of adolescents show a willingness to participate in obesity prevention programs. Participants suggest that any school-based program be voluntary, entertaining, informative, involve parents, take into consideration peer influence, and increase healthy food offerings while not eliminating individual choice. An ecological, school-based obesity prevention program with parent involvement should be developed for this adolescent population

Nivolumab-induced fulminant diabetic ketoacidosis followed by thyroiditis

Five days following the 3rd cycle of nivolumab, a monoclonal antibody, which acts as immune checkpoint inhibitor against the programmed cell death protein-1, for metastatic lung adenocarcinoma, a 56-year-old woman presented at the hospital critically ill. On admission, she had severe diabetic ketoacidosis (DKA), as evidenced by venous glucose of 47 mmol/L, blood ketones of 7.5 mmol/L, pH of 6.95 and bicarbonate of 6.6 mmol/L. She has had no personal or family history of diabetes mellitus (DM), while random venous glucose, measured 1 week prior to hospitalisation, was 6.1 mmol/L. On admission, her HbA1c was 8.2% and anti-GAD antibodies were 12 kIU/L (0–5 kU/L), while islet cell antibodies and serum C-peptide were undetectable. Nivolumab was recommenced without the development of other immune-mediated phenomena until 6 months later, when she developed hypothyroidism with TSH 18 U/L and low free T4. She remains insulin dependent and has required levothyroxine replacement, while she has maintained good radiological and clinical response to immunotherapy. This case is notable for the rapidity of onset and profound nature of DKA at presentation, which occurred two months following commencement of immunotherapy. Despite the association of nivolumab with immune-mediated endocrinopathies, only a very small number of patients developing type 1 DM has been reported to date. Patients should be closely monitored for hyperglycaemia and thyroid dysfunction prior to and periodically during immunotherapy

A Chemical and Enzymatic Approach to Study Site-Specific Sumoylation.

A variety of cellular pathways are regulated by protein modifications with ubiquitin-family proteins. SUMO, the Small Ubiquitin-like MOdifier, is covalently attached to lysine on target proteins via a cascade reaction catalyzed by E1, E2, and E3 enzymes. A major barrier to understanding the diverse regulatory roles of SUMO has been a lack of suitable methods to identify protein sumoylation sites. Here we developed a mass-spectrometry (MS) based approach combining chemical and enzymatic modifications to identify sumoylation sites. We applied this method to analyze the auto-sumoylation of the E1 enzyme in vitro and compared it to the GG-remnant method using Smt3-I96R as a substrate. We further examined the effect of smt3-I96R mutation in vivo and performed a proteome-wide analysis of protein sumoylation sites in Saccharomyces cerevisiae. To validate these findings, we confirmed several sumoylation sites of Aos1 and Uba2 in vivo. Together, these results demonstrate that our chemical and enzymatic method for identifying protein sumoylation sites provides a useful tool and that a combination of methods allows a detailed analysis of protein sumoylation sites

Novel DNA-based microfluorimetric method to evaluate antimalarial drug activity

Journal ArticleThis paper describes the development of a novel microfluorimetric assay to measure the inhibition of Plasmodium falciparum based on the detection of parasitic DNA by intercalation with PicoGreen®. The method was used to determine parasite inhibition profiles and 50% inhibitory concentration values of known or potential antimalarial drugs. Values for parasite inhibition with known anti-malarial drugs using the PicoGreen® assay were comparable with those determined by the standard method based upon the uptake of 3H-hypoxanthine and the Giemsa stain microscopic technique

Migrating Faculty Publications from a Department Website to an Institutional Repository

In October 2013, George Washington University\u27s School of Public Health and Health Services (now the Milken Institute School of Public Health) approached the Himmelfarb Health Sciences Library about moving a large collection of Health Policy Department publications and citations from the school\u27s website into the Health Sciences Research Commons (HSRC) institutional repository. After a project planning meeting with the school\u27s representative, library staff began uploading documents and citations from the collection. Working from a spreadsheet and file set provided by the school, library staff processed 604 citations (including 319 full-text files) dating back to 1996. Despite some problem files and citations, the majority were added to HSRC between November 2013 and April 2014. The successful completion of this project led to increased exposure of faculty research through standardized metadata, search engine optimization, and inclusion of research in the Digital Commons Network. Additionally, the establishment of new partnerships between the library and the public health school opened the door to future scholarly communications collaborations

A persistent and dynamic East Greenland Ice Sheet over the past 7.5 million years

Climate models show that ice-sheet melt will dominate sea-level rise over the coming centuries, but our understanding of ice-sheet variations before the last interglacial 125,000 years ago remains fragmentary. This is because terrestrial deposits of ancient glacial and interglacial periods1,2,3 are overrun and eroded by more recent glacial advances, and are therefore usually rare, isolated and poorly dated4. In contrast, material shed almost continuously from continents is preserved as marine sediment that can be analysed to infer the time-varying state of major ice sheets. Here we show that the East Greenland Ice Sheet existed over the past 7.5 million years, as indicated by beryllium and aluminium isotopes (10Be and 26Al) in quartz sand removed by deep, ongoing glacial erosion on land and deposited offshore in the marine sedimentary record5,6. During the early Pleistocene epoch, ice cover in East Greenland was dynamic; in contrast, East Greenland was mostly ice-covered during the mid-to-late Pleistocene. The isotope record we present is consistent with distinct signatures of changes in ice sheet behaviour coincident with major climate transitions. Although our data are continuous, they are from low-deposition-rate sites and sourced only from East Greenland. Consequently, the signal of extensive deglaciation during short, intense interglacials could be missed or blurred, and we cannot distinguish between a remnant ice sheet in the East Greenland highlands and a diminished continent-wide ice sheet. A clearer constraint on the behaviour of the ice sheet during past and, ultimately, future interglacial warmth could be produced by 10Be and 26Al records from a coring site with a higher deposition rate. Nonetheless, our analysis challenges the possibility of complete and extended deglaciation over the past several million years