Development of a multiplex real-time quantitative reverse-transcription polymerase chain reaction for the detection of four bee viruses.

Viruses in the families Dicistroviridae and Iflaviridae are among the main threats to western honey bees (Apis mellifera) and native bee species. Polymerase chain reaction (PCR) is the gold standard for pathogen detection in bees. However, high throughput screening for bee virus infections in singleplex PCR reactions is cumbersome and limited by the high quantities of sample RNA required. Thus, the development of a sensitive and specific multiplex PCR detection method for screening for multiple viruses simultaneously is necessary. Here, we report the development of a one-step multiplex reverse-transcription quantitative polymerase chain reaction (RT-qPCR) assay to detect four viruses commonly encountered in pollinator species. The optimized multiplex RT-qPCR protocol described in this study allows simultaneous detection of two dicistroviruses (Israeli acute paralysis virus and Black queen cell virus) and two iflaviruses (Sacbrood virus and Deformed wing virus) with high efficiency and specificity comparable to singleplex detection assays. This assay provides a broad range of detection and quantification, and the results of virus quantification in this study are similar to those performed in other studies using singleplex detection assays. This method will be particularly useful for data generation from small-bodied insect species that yield low amounts of RNA.

Synthesis and catalytic activity of single-site group V alkoxide complexes for the ring-opening polymerization of ε-caprolactone.

The synthesis, characterization, and ring-opening polymerization (ROP) activity of a family of niobium and tantalum alkoxide catalysts was studied. The final catalysts are made in a two-step synthesis, first by reacting the desired homoleptic metal ethoxide with a phenolketoimine ligand to form a series of synthetic intermediates, followed by reaction with catechol to produce a catalytic platform with a single ethoxide initiator. By using two separate ligands, the electronic properties of the catalyst can be tuned, and the molecular weight of the polymer can be increased. It was found that synthetic intermediates adopted a mer geometry both in solution and in the solid state. This mer geometry was retained for the final catechol derivatives, however in one case, where catechol was substituted for 3-methoxycatechol, the molecule adopted a highly distorted fac geometry. Catalytic ROP activity of the synthetic intermediates and final catechol derivatives with ε-caprolactone was studied through a kinetic analysis. In all seven cases studied the reactions proceeded through the expected coordination-insertion mechanism, following pseudo first-order kinetics and increasing in Mn linearly vs. conversion. The single-initiator catechol derivatives increased the Mn by three times compared to that of the three-initiator synthetic intermediates with little decrease in the overall reaction rate. Both the nature of the ligand and metal were found to impact the rate of reaction in these systems. By switching from an electron donating ligand to an electron withdrawing ligand, the rate was found to nearly double. Tantalum species were faster than their niobium counterparts by ∼3 times in the synthetic intermediates and ∼1.5 times in the catechol derivatives. This observed periodicity supports recent literature findings in this area.

Prevalence and effects of a parasitic trematode on the blue mussel, Mytilus edulis, in the Boston Harbor.

The blue mussel, Mytilus edulis, is a keystone species in the North Atlantic that plays critical roles in nutrient cycling, water filtration, and habitat creation. Blue mussel populations have declined significantly throughout the North Atlantic due to various factors, including habitat loss, pollution, increasing water temperature, and parasites. One parasite is Proctoeces maculatus, a digenetic trematode, which uses M. edulis as an intermediate host. This parasite causes reduced growth, castration, and death in mussels. The range of P. maculatus has expanded northward from Cape Cod, MA to Maine which may be associated with rising temperatures in the Gulf of Maine. To evaluate the negative impacts of P. maculatus on mussels, we analyzed its infections in M. edulis throughout the Boston Harbor, MA. P. maculatus was present in every population and time point analyzed, with approximately 50% of mussels in the harbor infected. The parasite reduced gonadal development in infected mussels, which could lead to decreased fecundity. Severe P. maculatus infections induced a stress response, indicated by increased HSP70 expression. We developed a non-destructive hemolymph-based assay to determine if mussels are infected with P. maculatus, thus speeding up the evaluation process and eliminating the need to sacrifice individuals. With P. maculatus' continued expansion northward, more mussel populations will be under threat from this parasite.

Continuous antibiotic prophylaxis in isolated prenatal hydronephrosis.

BACKGROUND: Prenatal hydronephrosis (PNH) is one of the most common congenital anomalies and can increase the risk of developing a urinary tract infection (UTI) in the first two years of life. Continuous antibiotic prophylaxis (CAP) has been recommended empirically to prevent UTI in children with PNH, but its use has been controversial. OBJECTIVE: We describe the incidence of UTI in children with isolated PNH of the renal pelvis without ureteral dilation. Our objective was to compare patients receiving and not receiving CAP and determine whether CAP is beneficial at preventing UTI in children with isolated PNH. STUDY DESIGN: Children with confirmed PNH were enrolled between 2008 and 2020 into the Society for Fetal Urology Hydronephrosis Registry. Children with isolated dilation of the renal pelvis without ureteral or bladder abnormality were included. The primary outcome was development of a UTI, comparing patients who were prescribed and not prescribed CAP. RESULTS: In this cohort of 801 children, 76% were male, and 35% had high grade hydronephrosis (SFU grades 3-4). CAP was prescribed in 34% of children. The UTI rate among all children with isolated PNH was 4.2%. Independent predictors of UTI were female sex (HR = 13, 95% CI: 3.8-40, p = 0.0001), intact prepuce (HR = 5.1, 95% CI: 1.4-18, p = 0.01) and high grade hydronephrosis (HR = 2.0, 95% CI: 0.99-4.0, p = 0.05; Table) on multivariable analysis. For patients on CAP, the UTI rate was 4.0% compared to 4.3% without CAP (p = 0.76). The risk of UTI during follow-up was not significantly different between patients who received CAP and patients who were not exposed to CAP; adjusting for sex, circumcision status and hydronephrosis grade (HR = 0.72, 95% CI: 0.34-1.5, p = 0.38). In sub-group analysis of patients at higher risk of UTI (uncircumcised males, females and high grade hydronephrosis), CAP use was not associated with a statistically significant reduction in UTI. CONCLUSIONS: The overall UTI rate in children with isolated PNH is very low at 4.2%. In the overall population of patients with isolated PNH, CAP was not associated with reduction in UTI risk, although the limitations in our study make characterizing CAP effectiveness difficult. Clinicians should consider risk factors prior to placing all patients with isolated PNH on CAP.

We Don't Have a Lot of Healthy Options: Food Environment Perceptions of First-Year, Minority College Students Attending a Food Desert Campus.

First-year college students are at particular risk of dietary maladaptation during their transition to adulthood. A college environment that facilitates consistent access to nutritious food is critical to ensuring dietary adequacy among students. The objective of the study was to examine perceptions of the campus food environment and its influence on the eating choices of first-year students attending a minority-serving university located in a food desert. Focus group interviews with twenty-one first-year students were conducted from November 2016 to January 2017. Students participated in 1 of 5 focus groups. Most interviewees identified as being of Hispanic/Latino or Asian/Pacific Islander origin. A grounded theory approach was applied for inductive identification of relevant concepts and deductive interpretation of patterns and relationships among themes. Themes related to the perceived food environment included adequacy (i.e., variety and quality), acceptability (i.e., familiarity and preferences), affordability, and accessibility (i.e., convenience and accommodation). Subjective norms and processes of decisional balance and agency were themes characterizing interpersonal and personal factors affecting students' eating choices. The perceived environment appeared to closely interact with subjective norms to inform internal processes of decision-making and agency around the eating choices of first-year students attending a minority-serving university campus located in a food desert.

Glucoregulatory and Cardiometabolic Profiles of Almond vs. Cracker Snacking for 8 Weeks in Young Adults: A Randomized Controlled Trial.

The transition to nutritional independence makes new college students vulnerable to alterations in eating patterns, which can increase the risk of cardiometabolic disorders. The aim of the study was to examine the potential benefits of almond vs. cracker snacking in improving glucoregulatory and cardiometabolic profiles in new college students. A randomized controlled, parallel-arm, 8-week intervention of 73 college students (BMI: 18⁻41 kg/m²) with no cardiometabolic disorders was conducted. Participants were randomized into either an almond snack group (56.7 g/day; 364 kcal; n = 38) or Graham cracker control group (77.5 g/day; 338 kcal/d; n = 35). Chronic, static changes were assessed from fasting serum/plasma samples at baseline, and after 4 and 8 weeks. Acute, dynamic effects were assessed during a 2-h oral glucose tolerance test (OGTT) at 8 weeks. Almond snacking resulted in a smaller decline in HDL cholesterol over 8 weeks (13.5% vs. 24.5%, p < 0.05), 13% lower 2-h glucose area under the curve (AUC), 34% lower insulin resistance index (IRI) and 82% higher Matsuda index (p < 0.05) during the OGTT, despite similar body mass gains over 8 weeks compared with the cracker group. In general, both almond and cracker snacking reduced fasting glucose, and LDL cholesterol. CONCLUSIONS: Incorporating a morning snack in the dietary regimen of predominantly breakfast-skipping, first-year college students had some beneficial effects on glucoregulatory and cardiometabolic health. Almond consumption has the potential to benefit postprandial glucoregulation in this cohort. These responses may be influenced by cardiometabolic risk factor status.

A tethered delivery mechanism explains the catalytic action of a microtubule polymerase.

Stu2p/XMAP215 proteins are essential microtubule polymerases that use multiple αß-tubulin-interacting TOG domains to bind microtubule plus ends and catalyze fast microtubule growth. We report here the structure of the TOG2 domain from Stu2p bound to yeast αß-tubulin. Like TOG1, TOG2 binds selectively to a fully 'curved' conformation of αß-tubulin, incompatible with a microtubule lattice. We also show that TOG1-TOG2 binds non-cooperatively to two αß-tubulins. Preferential interactions between TOGs and fully curved αß-tubulin that cannot exist elsewhere in the microtubule explain how these polymerases localize to the extreme microtubule end. We propose that these polymerases promote elongation because their linked TOG domains concentrate unpolymerized αß-tubulin near curved subunits already bound at the microtubule end. This tethering model can explain catalyst-like behavior and also predicts that the polymerase action changes the configuration of the microtubule end.