The use of molecular scatology to study river otter (Lontra canadensis) genetics

North American river otters (Lontra canadensis) were extirpated throughout all of Western New York due to habitat loss, pollution, and trapping. Between 1995 and 2000, 279 river otters were released throughout Western New York, 31 of which were released in the Genesee river watershed. Since their release there have been no followup studies on the river otters until the RIT River Otter Lab was formed in 2004. Researchers surveyed three local creeks to record data on toilet site locations and collect otter feces in order to perform dietary and genetic analyses. Through the use of molecular scatology I extracted DNA from feces in order to determine the amount of genetic diversity of the reintroduced river otter population. I also utilized otter scat samples from British Columbia and the Thousand Islands. Using a QIAGEN QIAamp Stool Mini Kit I attempted to extract mitochondrial cytochrome b DNA from 177 samples, roughly 16% of which were successfully amplified and sequenced. From the sequenced scat samples I identified two otter, 14 raccoon, one beaver, one coyote, and three fish: common carp, golden redhorse, and shorthead redhorse from the Genesee watershed. I have also sequenced one sample as otter and one sample as pink salmon from British Columbia and five samples as bullhead catfish from the Thousand Islands. It is believed that the samples that were sequenced as fish were likely from otters. I then utilized microsatellites, and I included a raccoon sample as well. To my surprise the raccoon sample worked with the river otter microsatellite primer, despite a 25% divergence between the two species’ cytochrome b sequences. I determined that out of ten river otter microsatellite primers: three river otter primers do not work with raccoons, five primers produced identical or nearly identical sequence, and two primers need more research to determine if they work with raccoons. These results stress the importance of confirming species identification from fecal samples using mitochondrial DNA prior to the use of microsatellites to avoid misleading results

Postpartum sterilization choices made by HIV-infected women.

OBJECTIVE: To assess if HIV-infected women made different choices for postpartum sterilization after implementation of the Pediatric AIDS Clinical Trials Group protocol 076 (November 1, 1994) compared to before implementation. STUDY DESIGN: A retrospective cohort study in which medical records were reviewed to obtain demographic, obstetric and HIV-related data from January 1993 through December 2002. HIV-infected women who completed a pregnancy by birth or abortion were divided into two comparison groups: "Pre-076" and "Post-076". The primary outcome was sterilization by postpartum tubal ligation.Results. Forty-two women (74%) in the Pre-076 group chose sterilization compared to 139 of 310 women (45%) in the Post-076 group (unadjusted OR 3.44, 95% CI 1.83, 6.47). Seventy-one percent of women younger than 21 years of age in the Pre-076 Group chose sterilization compared with only 35% of women younger than 21 years in the Post-076 group (p = 0.0136). Similarly, 78% of primiparous women chose sterilization after their first pregnancy in the Pre-076 group, compared to 14% in the Post-076 group (p < 0.001). CONCLUSIONS: Since the implementation of PACTG 076 protocol in November 1994, fewer HIV-infected women chose postpartum sterilization. The typical woman who now chooses postpartum sterilization is less likely to be young or primiparous than those who chose sterilization before PACTG Protocol 076 implementation

COVID-19 symptoms at hospital admission vary with age and sex: results from the ISARIC prospective multinational observational study

Background: The ISARIC prospective multinational observational study is the largest cohort of hospitalized patients with COVID-19. We present relationships of age, sex, and nationality to presenting symptoms. Methods: International, prospective observational study of 60 109 hospitalized symptomatic patients with laboratory-confirmed COVID-19 recruited from 43 countries between 30 January and 3 August 2020. Logistic regression was performed to evaluate relationships of age and sex to published COVID-19 case definitions and the most commonly reported symptoms. Results: ‘Typical’ symptoms of fever (69%), cough (68%) and shortness of breath (66%) were the most commonly reported. 92% of patients experienced at least one of these. Prevalence of typical symptoms was greatest in 30- to 60-year-olds (respectively 80, 79, 69%; at least one 95%). They were reported less frequently in children (≤ 18 years: 69, 48, 23; 85%), older adults (≥ 70 years: 61, 62, 65; 90%), and women (66, 66, 64; 90%; vs. men 71, 70, 67; 93%, each P &lt; 0.001). The most common atypical presentations under 60 years of age were nausea and vomiting and abdominal pain, and over 60 years was confusion. Regression models showed significant differences in symptoms with sex, age and country. Interpretation: This international collaboration has allowed us to report reliable symptom data from the largest cohort of patients admitted to hospital with COVID-19. Adults over 60 and children admitted to hospital with COVID-19 are less likely to present with typical symptoms. Nausea and vomiting are common atypical presentations under 30 years. Confusion is a frequent atypical presentation of COVID-19 in adults over 60 years. Women are less likely to experience typical symptoms than men

The interaction of the HSV-1 tegument proteins pUL36 and pUL37 is essential for secondary envelopment during viral egress

The herpes simplex virus type 1 (HSV-1) tegument proteins pUL36 (VP1/2) and pUL37 are essential for viral egress. We previously defined a minimal domain in HSV-1 pUL36, residues 548–572, as important for binding pUL37. Here, we investigated the role of this region in binding to pUL37 and facilitating viral replication. We deleted residues 548–572 in frame in a virus containing a mRFP tag at the N-terminus of the capsid protein VP26 and an eGFP tag at the C-terminus of pUL37 (HSV-1pUL36∆548-572). This mutant virus was unable to generate plaques in Vero cells, indicating that deletion of this region of pUL36 blocks viral replication. Imaging of HSV-1pUL36∆548-572-infected Vero cells, in comparison to parental and resucant, revealed a block in secondary envelopment of cytoplasmic capsids. In addition, immunoblot analysis suggested that failure to bind pUL37 affected the stability of pUL36. This study provides further insight into the role of this essential interaction

Muc5b is required for airway defence

Respiratory surfaces are exposed to billions of particulates and pathogens daily. A protective mucus barrier traps and eliminates them through mucociliary clearance (MCC). However, excessive mucus contributes to transient respiratory infections and to the pathogenesis of numerous respiratory diseases. MUC5AC and MUC5B are evolutionarily conserved genes that encode structurally related mucin glycoproteins, the principal macromolecules in airway mucus. Genetic variants are linked to diverse lung diseases, but specific roles for MUC5AC and MUC5B in MCC, and the lasting effects of their inhibition, are unknown. Here we show that mouse Muc5b (but not Muc5ac) is required for MCC, for controlling infections in the airways and middle ear, and for maintaining immune homeostasis in mouse lungs, whereas Muc5ac is dispensable. Muc5b deficiency caused materials to accumulate in upper and lower airways. This defect led to chronic infection by multiple bacterial species, including Staphylococcus aureus, and to inflammation that failed to resolve normally. Apoptotic macrophages accumulated, phagocytosis was impaired, and interleukin-23 (IL-23) production was reduced in Muc5b(-/-) mice. By contrast, in mice that transgenically overexpress Muc5b, macrophage functions improved. Existing dogma defines mucous phenotypes in asthma and chronic obstructive pulmonary disease (COPD) as driven by increased MUC5AC, with MUC5B levels either unaffected or increased in expectorated sputum. However, in many patients, MUC5B production at airway surfaces decreases by as much as 90%. By distinguishing a specific role for Muc5b in MCC, and by determining its impact on bacterial infections and inflammation in mice, our results provide a refined framework for designing targeted therapies to control mucin secretion and restore MCC