A stem-cell-derived platform enables complete Cryptosporidium development in vitro and genetic tractability

Despite being a frequent cause of severe diarrheal disease in infants and an opportunistic infection in immunocompromised patients, Cryptosporidium research has lagged due to a lack of facile experimental methods. Here, we describe a platform for complete life cycle development and long-term growth of C. parvum in vitro using air-liquid interface (ALI) cultures derived from intestinal epithelial stem cells. Transcriptomic profiling revealed that differentiating epithelial cells grown under ALI conditions undergo profound changes in metabolism and development that enable completion of the parasite life cycle in vitro. ALI cultures support parasite expansion \u3e 100-fold and generate viable oocysts that are transmissible in vitro and to mice, causing infection and animal death. Transgenic parasite lines created using CRISPR/Cas9 were used to complete a genetic cross in vitro, demonstrating Mendelian segregation of chromosomes during meiosis. ALI culture provides an accessible model that will enable innovative studies into Cryptosporidium biology and host interactions

The Bulk RS KK-gluon at the LHC

We study the possibility of discovering and measuring the properties of the lightest Kaluza-Klein excitation of the gluon in a Randall-Sundrum scenario where the Standard Model matter and gauge fields propagate in the bulk. The KK-gluon decays primarily into top quarks. We discuss how to use the $t \bar{t}$ final states to discover and probe the properties of the KK-gluon. Identification of highly energetic tops is crucial for this analysis. We show that conventional identification methods relying on well separated decay products will not work for heavy resonances but suggest alternative methods for top identification for energetic tops. We find, conservatively, that resonances with masses less than 5 TeV can be discovered if the algorithm to identify high $p_T$ tops can reject the QCD background by a factor of 10. We also find that for similar or lighter masses the spin can be determined and for lighter masses the chirality of the coupling to $t\bar t$ can be measured. Since the energetic top pair final state is a generic signature for a large class of new physics as the top quark presumably couples most strongly to the electroweak symmetry breaking sector, the methods we have outlined to study the properties of the KK-gluon should also be important in other scenarios.Comment: 21 pages, 13 figure

Effects of Art Intervention on Pediatric Anxiety and Pain in the Medical Setting

Introduction: Hospitalization and illness can be a painful and stressful time for a child. There may be anxiety over procedures and inpatient stays disrupt normal routines. Previous research found that for pre-school aged children, having parents around, having the help of the hospital staff, and playing an active role in alleviating their fears were the most helpful in reducing anxiety. Another study found that visual creative expressions can be meaningful experiences for young adult cancer survivors. Additionally, there is abundant literature on formal art therapy and its favorable effects on children in the hospital, however, there are fewer studies investigating less standardized “art intervention” in the same population. The purpose of our project was to assess whether art intervention reduces anxiety and pain in inpatient and outpatient pediatric patients.https://scholarworks.uvm.edu/comphp_gallery/1224/thumbnail.jp

Rectal microbicides: clinically relevant approach to the design of rectal specific placebo formulations

<p>Abstract</p> <p>Background</p> <p>The objective of this study is to identify the critical formulation parameters controlling distribution and function for the rectal administration of microbicides in humans. Four placebo formulations were designed with a wide range of hydrophilic characteristics (aqueous to lipid) and rheological properties (Newtonian, shear thinning, thermal sensitive and thixotropic). Aqueous formulations using typical polymers to control viscosity were iso-osmotic and buffered to pH 7. Lipid formulations were developed from lipid solvent/lipid gelling agent binary mixtures. Testing included pharmaceutical function and stability as well as <it>in vitro </it>and <it>in vivo </it>toxicity.</p> <p>Results</p> <p>The aqueous fluid placebo, based on poloxamer, was fluid at room temperature, thickened and became shear thinning at 37°C. The aqueous gel placebo used carbopol as the gelling agent, was shear thinning at room temperature and showed a typical decrease in viscosity with an increase in temperature. The lipid fluid placebo, myristyl myristate in isopropyl myristate, was relatively thin and temperature independent. The lipid gel placebo, glyceryl stearate and PEG-75 stearate in caprylic/capric triglycerides, was also shear thinning at both room temperature and 37°C but with significant time dependency or thixotropy. All formulations showed no rectal irritation in rabbits and were non-toxic using an <it>ex vivo </it>rectal explant model.</p> <p>Conclusions</p> <p>Four placebo formulations ranging from fluid to gel in aqueous and lipid formats with a range of rheological properties were developed, tested, scaled-up, manufactured under cGMP conditions and enrolled in a formal stability program. Clinical testing of these formulations as placebos will serve as the basis for further microbicide formulation development with drug-containing products.</p

Effect of a Low Carbohydrate-Moderate Protein Supplement on Endurance Performance in Female Athletes

Previous research has shown that consuming a carbohydrate supplement during prolonged endurance exercise improves performance compared to water or placebo. The addition of protein to traditional carbohydrate supplement has been shown to further improve perfomance beyond that of carbohydrate alone. However, few investigations have explored the effect of adding protein to a supplement containing a low carbohydrate concentration. PURPOSE: To investigate if a low carbohydrate and moderate protein supplement, provided during prolonged variable intensity exercise, would improve time to exhaustion in comparison to a traditional carbohydrate supplement. METHODS: Fourteen (n = 14) trained females cyclists and triathletes (30.4 ±1.6 yrs, 2.90 ± 0.15 L⋅min¬-1) cycled on two different occasions for three hours at intensities varying between 45% - 70% VO2max. After three hours, the intensity was increased (average 72.5 % VO2max) and held until exhaustion. Exhaustion was defined as the point at which subjects could no longer hold cadence above 60RPM. Supplements (275ml) were provided every 20 min during exercise and were composed of a 3% carbohydrate/1.2% protein mix (CHO+PRO) or a 6% carbohydrate-only (CHO). The CHO+PRO treatment contained a mixture of glucose (dextrose), maltodextrin and fructose, and whey protein isolate. The CHO treatment was composed of dextrose. CHO+PRO contained half the carbohydrate content and 30% less calories in comparison to CHO. RESULTS: Time to exhaustion (TTE) was significantly greater with CHO+PRO in comparison to CHO (49.94 ± 7.01 vs 42.36 ± 6.21 min, respectively, p = 0.039). CONCLUSIONS: The above result suggests that addition of a moderate protein to a low carbohydrate supplement enhances performance in endurance trained females above that of carbohydrate alone. Improvement in performance occurred despite a lower carbohydrate and caloric content. It is unknown whether the greater performance seen with CHO+PRO was a result of the added protein, the use of a mixture of carbohydrate sources (glucose, maltodextrin and fructose), or their combination

Distribution of the

OBJECTIVE: To study whether reported, but inconsistent, associations between the FMR1 CGG repeat lengths in the intermediate, high normal, or low normal range differentiate women diagnosed with diminished ovarian reserve (DOR) from population controls and whether associations vary by race/ethnic group. DESIGN: Case-control study. SETTING: Academic and private fertility clinics. PATIENT(S): DOR cases (n = 129; 95 Whites, 22 Asian, 12 other) from five U.S. fertility clinics were clinically diagnosed, with regular menses and no fragile X syndrome family history. Normal fertility controls (n = 803; 386 Whites, 219 African-Americans, 102 Japanese, 96 Chinese) from the United States-based SWAN Study had one or more menstrual period in the 3 months pre-enrollment, one or more pregnancy, no history of infertility or hormone therapy, and menopause ≥46 years. Previously, the SWAN Chinese and Japanese groups had similar FMR1 CGG repeat lengths, thus they were combined. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): FMR1 CGG repeat lengths. RESULT(S): Median CGG repeats were nearly identical by case/control group. DOR cases had fewer CGG repeats in the shorter FMR1 allele than controls among Whites, but this was not significant among Asians. White cases had fewer CGG repeats in the shorter allele than Asian cases. No significant differences were found in the high normal/intermediate range between cases and controls or by race/ethnic group within cases in the longer allele. CONCLUSION(S): This study refutes prior reports of an association between DOR and high normal/intermediate repeats and confirms an association between DOR and low normal repeats in Whites

Population dynamics of rhesus macaques and associated foamy virus in Bangladesh.

Foamy viruses are complex retroviruses that have been shown to be transmitted from nonhuman primates to humans. In Bangladesh, infection with simian foamy virus (SFV) is ubiquitous among rhesus macaques, which come into contact with humans in diverse locations and contexts throughout the country. We analyzed microsatellite DNA from 126 macaques at six sites in Bangladesh in order to characterize geographic patterns of macaque population structure. We also included in this study 38 macaques owned by nomadic people who train them to perform for audiences. PCR was used to analyze a portion of the proviral gag gene from all SFV-positive macaques, and multiple clones were sequenced. Phylogenetic analysis was used to infer long-term patterns of viral transmission. Analyses of SFV gag gene sequences indicated that macaque populations from different areas harbor genetically distinct strains of SFV, suggesting that geographic features such as forest cover play a role in determining the dispersal of macaques and SFV. We also found evidence suggesting that humans traveling the region with performing macaques likely play a role in the translocation of macaques and SFV. Our studies found that individual animals can harbor more than one strain of SFV and that presence of more than one SFV strain is more common among older animals. Some macaques are infected with SFV that appears to be recombinant. These findings paint a more detailed picture of how geographic and sociocultural factors influence the spectrum of simian-borne retroviruses

Rheumatoid Arthritis Naive T Cells Share Hypermethylation Sites With Synoviocytes.

ObjectiveTo determine whether differentially methylated CpGs in synovium-derived fibroblast-like synoviocytes (FLS) of patients with rheumatoid arthritis (RA) were also differentially methylated in RA peripheral blood (PB) samples.MethodsFor this study, 371 genome-wide DNA methylation profiles were measured using Illumina HumanMethylation450 BeadChips in PB samples from 63 patients with RA and 31 unaffected control subjects, specifically in the cell subsets of CD14+ monocytes, CD19+ B cells, CD4+ memory T cells, and CD4+ naive T cells.ResultsOf 5,532 hypermethylated FLS candidate CpGs, 1,056 were hypermethylated in CD4+ naive T cells from RA PB compared to control PB. In analyses of a second set of CpG candidates based on single-nucleotide polymorphisms from a genome-wide association study of RA, 1 significantly hypermethylated CpG in CD4+ memory T cells and 18 significant CpGs (6 hypomethylated, 12 hypermethylated) in CD4+ naive T cells were found. A prediction score based on the hypermethylated FLS candidates had an area under the curve of 0.73 for association with RA case status, which compared favorably to the association of RA with the HLA-DRB1 shared epitope risk allele and with a validated RA genetic risk score.ConclusionFLS-representative DNA methylation signatures derived from the PB may prove to be valuable biomarkers for the risk of RA or for disease status