Analysis of qualitative and quantitative morphological traits related to yield in country bean (Lablab purpureus L. sweet) genotypes.

Country bean is a grain legume extensively farmed for its multi-purpose uses, yet the traits related to yield are are poorly studied and yet unexplored. A study on the diversity of qualitative and quantitative morphological characteristics concerning yield among the country bean germplasms collected from Bangladesh identified considerable variation in the studied traits across the germplasms and identified a complex correlation between the qualitative and quantitative traits. Principal Component Analysis (PCA) detected five components that contributed 66.38% qualitative traits and six components contributed 74.49% quantitative traits to total variations. Eigenvalues indicated that a majority of color-related qualitative traits included cotyledon, leaf, vein, seed, flower, and petals contributed, in contrast,a majority of the seed, leaf, flower, and inflorescence-related quantitative traits contributed to the total diversity of the Lablab germplasms. Among the quantitative traits, the highest coefficient of variation (CV%) was found in average pod weight (50.98%), followed by the total number of spikes per plant (43.82%), while seed length, pod weight, length, width, thickness, number of flower/spike, spike length, and total no of spikes/plant all had more than 20.00 percent CV, suggesting suitability to use in the breeding of high yielding genotypes. The germplasms are grouped into four and three clusters based on quantitative and qualitative traits, suggesting quantitative characters offer better clustering of genotypes. Considering the above traits, our research found that the BD-10804, BD-10807, BD-11091, BD-10808, BD-10815, and BD-11089 and cultivar Goal Goda Lablab beans germplasms produced higher pod weight with corresponding higher pod length, width, and thickness suggesting to use them as high yielding genotypes for food and fodder purposes.

Medical Student Attitudes Toward Blood Donation in Times of Increased Need.

BACKGROUND: In situations of increased need, such as mass casualty incidents (MCIs) and COVID-19, donated blood products are in shortage across the United States. Medical students are a potential pool for blood donors. The aim of this study was to determine overall attitudes of medical students at a single academic institution toward blood donation during times of increased need. METHODS: Three anonymous REDCap surveys were administered to all medical students at a rural academic institution. Surveys 1 and 2 were administered preceding and after an institution-wide MCI drill, in September and November 2019, respectively. Survey 3 was administered following a student-organized COVID-19 blood drive in June 2020. Multivariable analysis was performed to determine if factors, ie, experience with MCI drills and emergency medical services (EMS) training, were associated with willingness to donate blood. Furthermore, barriers to donation among those not willing to donate were assessed. RESULTS: Overall response rate for MCI surveys (surveys 1 and 2) was 38% (mean age 25.2 years and 50% women). 91% (n = 210) of respondents were willing to donate blood. Previous participation in MCI drills and EMS training was not associated with higher willingness to donate blood. Response rate for survey 3 was 15.6% (59.4% women), and 30 (31.3%) respondents indicated they did not volunteer to donate blood during the COVID-19 drive. Most common reasons for not donating were "other," medical concerns, and being out-of-town. CONCLUSIONS: Majority of medical students are willing to donate blood during times of increased need and offer a possible solution to increase blood donor pool.

Use of prescription drug monitoring program to audit opioid prescribing patterns for patients with symptomatic nephrolithiasis.

INTRODUCTION The opioid epidemic is a growing problem in the United States. There is a high rate of opioid oversupply for treatment of symptomatic nephrolithiasis, partly due to patients being seen by multiple providers. In Pennsylvania, there are efforts to integrate a prescription drug monitoring program (PDMP) within the electronic medical record (EMR). The objectives of this study were to evaluate prescribing practices for opioids for symptomatic nephrolithiasis and the incidence of prescriptions not documented within the EMR. MATERIALS AND METHODS: Adults who presented for treatment of symptomatic nephrolithiasis were sequentially evaluated from May - October 2017 at Penn State Milton S. Hershey Medical Center. With IRB approval, we evaluated opioids prescribed in the EMR, which was compared to the PDMP for each stone episode. We calculated daily morphine milligram equivalents (MME) and total MME available to patients. RESULTS: A total of 301 patients were identified (52% male) with a mean age of 50.0 +/- 16.7 years and 249 (83%) of patients were prescribed narcotics with an average of 226.8 +/- 232.2 MME for their stone episode. Of patients that were prescribed narcotics, 19% had additional narcotics prescribed to them that were not entered into the EMR and later identified using PDMP. The average additional opioid prescribed was 371.8 +/- 404.2 total MME. CONCLUSIONS: The majority of patients presenting with symptomatic nephrolithiasis were prescribed an opioid. Approximately one-fifth of patients were receiving opioids from other providers that were not documented in the EMR at the time of their opioid prescription. PDMP, or similar resources, should be utilized by providers to minimize opioid use and reduce oversupplying patients.

Patient Outcomes in Mesenteric Venous Thrombosis Treated With Empiric Antibiotics.

BACKGROUND: Mesenteric venous thrombosis (MVT) is typically associated with poor prognosis. Although prophylactic antibiotics are sometimes given with the intent of limiting bacterial luminal load and translocation in patients with MVT, this approach has not been universally adopted. The aim of this study is to analyze whether utilizing antibiotics empirically in those with MVT improves patient outcomes and survival when compared to those who do not receive empiric antibiotics. METHODS: A retrospective review of patients admitted with MVT between 2002 and 2019 at a single academic institution was performed. Demographics and rates of mortality need for bowel resection, readmission, and Clostridium difficile (C. diff) infection were compared between patients treated with empiric antibiotics and patients not treated with antibiotics. RESULTS: Eighty-three patients (mean age 64.5 years and 55.4% male) who were admitted for MVT were included. Empiric antibiotics were utilized in 53% (n = 44) of MVT patients while 47% (n = 39) received supportive treatment without empiric antibiotics. Death occurred in 4 patients treated with antibiotics and 6 patients treated without antibiotics (9.1% vs. 15.3%, P = .50). Readmissions occurred in 12 patients (27.3%) treated with antibiotics and 10 patients (25.6%) not treated with antibiotics (27.3% vs. 25.6%, P = .87). C. diff infection occurred in 6 patients treated with antibiotics and in no patients not treated with antibiotics (13.6% vs. 0%, P = .03). CONCLUSIONS: Empiric antibiotic usage may not improve rates of mortality or hospital readmission in patients with MVT and may unnecessarily expose patients to an increased risk of C. diff infection.

Neonatal monocytes demonstrate impaired homeostatic extravasation into a microphysiological human vascular model.

Infections are most frequent at the extremes of life, especially among newborns, reflecting age-specific differences in immunity. Monocytes maintain tissue-homeostasis and defence-readiness by escaping circulation in the absence of inflammation to become tissue-resident antigen presenting cells in vivo. Despite equivalent circulating levels, neonates demonstrate lower presence of monocytes inside peripheral tissues as compared to adults. To study the ability of monocytes to undergo autonomous transendothelial extravasation under biologically accurate circumstances we engineered a three-dimensional human vascular-interstitial model including collagen, fibronectin, primary endothelial cells and autologous untreated plasma. This microphysiological tissue construct enabled age-specific autonomous extravasation of monocytes through a confluent human endothelium in the absence of exogenous chemokines and activation. Both CD16- and CD16+ newborn monocytes demonstrated lower adherence and extravasation as compared to adults. In contrast, pre-activated tissue constructs were colonized by newborn monocytes at the same frequency than adult monocytes, suggesting that neonatal monocytes are capable of colonizing inflamed tissues. The presence of autologous plasma neither improved newborn homeostatic extravasation nor shaped age-specific differences in endothelial cytokines that could account for this impairment. Newborn monocytes demonstrated significantly lower surface expression of CD31 and CD11b, and mechanistic experiments using blocking antibodies confirmed a functional role for CD31 and CD54 in neonatal homeostatic extravasation. Our data suggests that newborn monocytes are intrinsically impaired in extravasation through quiescent endothelia, a phenomenon that could contribute to the divergent immune responsiveness to vaccines and susceptibility to infection observed during early life.

A Collaborative Response to Provocative Comments in a Medical School.

This article was migrated. The article was marked as recommended. In 2018, anonymous online provocative comments were submitted to student leaders of a Syrian Refugee Initiative (SRI) at the Penn State College of Medicine. This triggered a series of actions with students and medical school leaders aimed at identifying the person who submitted the comments, trying to understand mutual perspectives, and managing the impact of the event on the student body. We describe the history of our college's commitment to humanism and how the SRI was a direct outgrowth of that culture. Voices of the student leaders who were directly impacted by the provocative comments and educational leaders who worked to resolve the crisis are presented. We also describe a collaborative process that involved engaging cybersecurity experts to identify the perpetrator, and share how the students and educational leaders were able to develop trust despite initial skepticism by students over the leadership's avowed commitment to taking the hate speech seriously. While the perpetrator was never identified and opportunities for improvement were identified along the way, by including student leaders in the process, students and administrators were able to develop trust and reach reasonable closure on this disturbing event. Take home messages are presented to guide other institutions in navigating instances of provocative comments or speech.

27Al and 31P NMR spectroscopy method development to quantify aluminum phosphate in adjuvanted vaccine formulations.

A novel qNMR method is described for the quantitative determination of total aluminum and phosphate in aluminum phosphate (AlPO4) adjuvanted vaccine samples using solution 27Al and 31P nuclear magnetic resonance (NMR) spectroscopy. External standard calibrations of AlPO4 solutions established excellent linearity in the range of 15-40 × 10-3 M and additional studies determined the level of detection for both nuclei. A commercialized combination vaccine product (Quadracel®), along with several individual adsorbed antigen components used in the vaccine were employed as model systems for method development. The developed method is also capable of quantitating the free phosphate (i.e. the fraction not bound to AlPO4 particles) in adjuvanted vaccines. This study is the first demonstration of a solution NMR method that is suitable for measuring total aluminum, and free and total phosphate concentrations in vaccine formulations consisting of antigen(s) adsorbed to aluminum adjuvant, in a single analytical workflow.

Nuclear Magnetic Resonance (NMR) Study for the Detection and Quantitation of Cholesterol in HSV529 Therapeutic Vaccine Candidate.

This study describes the NMR-based method to determine the limit of quantitation (LOQ) and limit of detection (LOD) of cholesterol, a process-related impurity in the replication-deficient Herpes Simplex Virus (HSV) type 2 candidate vaccine HSV529. Three signature peaks from the 1D 1H NMR of a cholesterol reference spectrum were selected for the identification of cholesterol. The LOQ for a cholesterol working standard was found to be 1 µg/mL, and the LOD was found to be 0.1 µg/mL. The identity of cholesterol, separated from the formulation of growth supplement by thin layer chromatography (TLC), was confirmed by 1D 1H NMR and 2D 1H-13C HSQC NMR. The three signature peaks of cholesterol were detected only in a six-times concentrated sample of HSV529 candidate vaccine sample and not in the single dose HSV529 vaccine sample under similar experimental conditions. Taken together, the results demonstrated that NMR is a direct method that can successfully identify and quantify cholesterol in viral vaccine samples, such as HSV529, and as well as in the growth supplement used during the upstream stages of HSV529 manufacturing.

Enhanced Binding Affinity via Destabilization of the Unbound State: A Millisecond Hydrogen-Deuterium Exchange Study of the Interaction between p53 and a Pleckstrin Homology Domain.

The incorporation of intrinsically disordered domains enables proteins to engage a wide variety of targets, with phosphorylation often modulating target specificity and affinity. Although phosphorylation can clearly act as a chemical driver of complexation in structured proteins, e.g., by abrogating or permitting new charge-charge interactions, the basis for enhancement of the hydrophobically driven interactions that are typical of disordered protein-target complexation is less clear. To determine how phosphorylation can positively impact target recruitment in disordered domains, we have examined the interaction between the disordered N-terminal transactivation domain (TAD) of p53 and the pleckstrin homology (PH) domain of p62. Using time-resolved electrospray ionization with hydrogen-deuterium exchange, we demonstrate that phosphorylation has little effect on the conformation of the p53 TAD when it is bound to the PH domain but instead increases the degree of conformational disorder in the unbound state. We propose that this increase in the degree of disorder creates a wider free energy gap between the free and bound states, providing a target-independent mechanism for enhanced binding when the phosphorylated and unphosphorylated p53-target complexes have similar free energies.

E3 ubiquitin ligase Mule targets ß-catenin under conditions of hyperactive Wnt signaling.

Wnt signaling, named after the secreted proteins that bind to cell surface receptors to activate the pathway, plays critical roles both in embryonic development and the maintenance of homeostasis in many adult tissues. Two particularly important cellular programs orchestrated by Wnt signaling are proliferation and stem cell self-renewal. Constitutive activation of the Wnt pathway resulting from mutation or improper modulation of pathway components contributes to cancer development in various tissues. Colon cancers frequently bear inactivating mutations of the adenomatous polyposis coli (APC) gene, whose product is an important component of the destruction complex that regulates ß-catenin levels. Stabilization and nuclear localization of ß-catenin result in the expression of a panel of Wnt target genes. We previously showed that Mule/Huwe1/Arf-BP1 (Mule) controls murine intestinal stem and progenitor cell proliferation by modulating the Wnt pathway via c-Myc. Here we extend our investigation of Mule's influence on oncogenesis by showing that Mule interacts directly with ß-catenin and targets it for degradation under conditions of hyperactive Wnt signaling. Our findings suggest that Mule uses various mechanisms to fine-tune the Wnt pathway and provides multiple safeguards against tumorigenesis.

Ubiquitin-specific protease 7 is a regulator of ubiquitin-conjugating enzyme UbE2E1.

Ubiquitin-specific protease 7 (USP7) is a deubiquitinating enzyme found in all eukaryotes that catalyzes the removal of ubiquitin from specific target proteins. Here, we report that UbE2E1, an E2 ubiquitin conjugation enzyme with a unique N-terminal extension, is a novel USP7-interacting protein. USP7 forms a complex with UbE2E1 in vitro and in vivo through the ASTS USP7 binding motif within its N-terminal extension in an identical manner with other known USP7 binding proteins. We show that USP7 attenuates UbE2E1-mediated ubiquitination, an effect that requires the N-terminal ASTS sequence of UbE2E1 as well as the catalytic activity of USP7. Additionally, USP7 is critical in maintaining the steady state levels of UbE2E1 in cells. This study reveals a new cellular mechanism that couples the opposing activities of the ubiquitination machinery and a deubiquitinating enzyme to maintain and modulate the dynamic balance of the ubiquitin-proteasome system.

Activation and localization of protein kinase C in Neurospora crassa.

The Neurosporacrassa protein kinase C (NPKC) is reported to be a regulator of light responsive genes. It phosphorylates the light receptor WC-1 and regulates the levels of the circadian clock protein FRQ and transcription of the light-induced albino-2 gene. In mammals, the conventional and novel isoforms of PKC are activated by diacylglycerol (DAG), which induces PKC translocation from the cytoplasm to membranes. To investigate the interaction of NPKC and DAG in Neurospora, we constructed a strain that expresses a PKC-GFP fusion protein. We found that NPKC localizes to growing tips and sub-apical plasma membrane in actively growing hyphae, and actively participates in septum development. NPKC is activated by exogenous DAG and phorbol esters, and translocates to the plasma membrane from the cytoplasm. We have previously reported that choline depletion of the chol-1 mutant of Neurospora increases DAG levels and lengthens the period of the circadian rhythm of conidiation. We have found that the activity of NPKC is rhythmic, and that NPKC levels are increased on choline depletion. However, over-expression of NPKC did not lengthen the conidiation period, indicating that PKC in Neurospora may not be responsible for the lengthened period in low choline cultures.