Genetic analysis of M94 of murine cytomegalovirus

The M94 gene of murine cytomegalovirus (MCMV) is one of the about 40 core genes of the subfamily of β-Herpesvirinae with unknown function. MCMV is utilised as an in vivo model for studying human cytomegalovirus (HCMV) due to the strict species specifity of HCMV and major homologies to MCMV. HCMV is an important human pathogen with worldwide distribution. Although infection of the immune competent host is usually clinically silent, infection of the foetus and the newborn causes severe organ damage and infection of immunocompromised patients induces life-threatening disease. M94 has not been studied intensely as a single gene before, but studies on its homologues in other Herpesviridae demonstrated expression as true late gene, presence in the virion, interaction of UL94 and UL99 in HCMV and capsid binding in α-Herpesvirinae. While UL94 in HCMV is essential, M94 homologues in α-Herpesvirinae are not essential, indicating an essential function of M94 conserved in β-Herpesvirinae but lost in α-Herpesvirinae. For the identification of this essential function, a library of random mutants was generated by a modified Tn7 transposon mutagenesis. The modifications in the transposon mutagenesis produced a library of high diversity and wide coverage containing 32,000 primary clones including stop and insertion mutants. The analysis of 613 clones by sequencing resulted in 399 unique mutants containing a correct 15 base pair (bp) insertion in M94. 116 stop and insertion mutants were individually reinserted into the viral genome and studied in the viral context. The primary analysis showed the ability of certain mutants to complement the M94 deletion, thereby identifying the essential regions in the M94 protein. Secondly, the M94 mutants were re-inserted as second gene and analysed for their inhibitory capacity of the M94 wild type (wt) functions to identify dominant negative (DN) mutants. The inhibitory mutants found were then verified as DN mutants by regulated expression in the virus context and by analysis of their specific phenotype induced by overexpression. Further analysis of the DN mutant in comparison to a M94 deletion mutant revealed no effect of M94 in viral cleavage-packaging, despite previous publications. Additional analysis showed a block in secondary envelopment for both the DN mutant and the deletion mutant and the determination of the spread deficient phenotype of the M94 deletion virus. Altogether these results constitute a new model for secondary envelopment in β-Herpesvirinae

Student Evaluation of Teaching: The Inequity of Faculty Scores in Online versus Face-to-Face Courses

Researchers have conducted a multitude of studies over the last century on Student Evaluations of Teaching (SET); however, very few have been conducted in the new digital age (Loveland, 2007). More work is in progress as researchers try to define the differences in student responses and thoughts about the online teaching environment. The unfortunate side of this can be the administrative decision to use a one-size-fits-all mentality when many authors including Dziuban and Moskal (2011) have outlined several research-based alternatives for evaluation of online instructional effectiveness. SETs are important to faculty because they often are the determining factor in merit pay and tenure/promotion. Faculty use SETs to guide decision-making about their curriculum and instructional strategies used to deliver the course (Sheehan & DuPrey, 1999). The power of the SET is recognized as a driving force in academia. What are often not discussed are the differences between SET for face-to-face courses versus SET for online courses

Perceptions of Violations by Artificial and Human Actors across Moral Foundations

Artificial agents such as robots, chatbots, and artificial intelligence systems can be the perpetrators of a range of moral violations traditionally limited to human actors. This paper explores how people perceive the same moral violations differently for artificial agent and human perpetrators by addressing three research questions: How wrong are moral foundation violations by artificial agents compared to human perpetrators? Which moral foundations do artificial agents violate compared to human perpetrators? What leads to increased blame for moral foundation violations by artificial agents compared to human perpetrators? We adapt 18 human-perpetrated moral violation scenarios that differ by the moral foundation violated (harm, unfairness, betrayal, subversion, degradation, and oppression) to create 18 agent-perpetrated moral violation scenarios. Two studies compare human-perpetrated to agent-perpetrated scenarios. They reveal that agent-perpetrated violations are more often perceived as not wrong or violating a different foundation than their human counterparts. People are less likely to classify violations by artificial agents as oppression and subversion, the foundations that deal the most with group hierarchy. Finally, artificial agents are blamed less than humans across moral foundations, and this blame is based more on the agent\u27s ability and intention for every moral foundation except harm

The Effects of Technology Integration Techniques in Elementary Mathematics Methods Courses on Elementary Preservice Teachers' Computer Self-Efficacy, Software Integration Confidence, and Lesson Planning

The purpose of this study was to demonstrate the effect of computer technology integration techniques on preservice teachers' feelings of computer self-efficacy and feelings of confidence in software integration. It was also the purpose of this study to interpret these preservice teachers' confidence in using computer technology integration techniques in their own planning and instruction during student teaching. The participants in this study were from two intact, non-randomly-formed classrooms. They were 27 preservice teachers enrolled in the College of Education at a university in north central Texas in two sections of a course entitled EDEE 4350, Mathematics in the Elementary School. This study was quasi-experimental, with a nonequivalent pretest-posttest control group design. The independent variable was the type of instruction experienced in an elementary mathematics methods course: novel instruction with specialized computer technology integration techniques versus traditional instruction with no specialized technology integration techniques. The dependant variables were measured using the following instruments: the Demographic Data and Previous Context Use of the Computer Survey which described participants' demographics and their previous usage of the computer; the Self-Efficacy With Computer Technologies Scale; the Preservice Teacher Software Integration Confidence Scale; and the Lesson Plan Infusion/Integration Scale. The results of the data analysis revealed, through the inferential statistics run on the Self-Efficacy with Computer Technology Scale pretest and posttest, that there was no statistically significant difference between treatment groups (p < .05). The posttest-only Preservice Teachers Software Integration Confidence Scale revealed a statistically significant difference between treatment groups (p < .05). The posttest-only Lesson Plan Technology Infusion/Integration Scale revealed no statistical significance between treatment groups (p < .05). The study provides insight into the benefits of instruction in specific software integration techniques instruction. It suggests that when preservice teachers are given instruction in specific computer software integration techniques, they are more confident in the use of those techniques

Downregulation of p53 drives autophagy during human trophoblast differentiation

The placental barrier is crucial for the supply of nutrients and oxygen to the developing fetus and is maintained by differentiation and fusion of mononucleated cytotrophoblasts into the syncytiotrophoblast, a process only partially understood. Here transcriptome and pathway analyses during differentiation and fusion of cultured trophoblasts yielded p53 signaling as negative upstream regulator and indicated an upregulation of autophagy-related genes. We further showed p53 mRNA and protein levels decreased during trophoblast differentiation. Reciprocally, autophagic flux increased and cytoplasmic LC3B-GFP puncta became more abundant, indicating enhanced autophagic activity. In line, in human first trimester placenta p53 protein mainly localized to the cytotrophoblast, while autophagy marker LC3B as well as late autophagic compartments were predominantly detectable in the syncytiotrophoblast. Importantly, ectopic overexpression of p53 reduced levels of LC3B-II, supporting a negative regulatory role on autophagy in differentiating trophoblasts. This was also shown in primary trophoblasts and human first trimester placental explants, where pharmacological stabilization of p53 decreased LC3B-II levels. In summary our data suggest that differentiation-dependent downregulation of p53 is a prerequisite for activating autophagy in the syncytiotrophoblast

Mindfulness Intervention for Stress Eating to Reduce Cortisol and Abdominal Fat among Overweight and Obese Women: An Exploratory Randomized Controlled Study

Psychological distress and elevated cortisol secretion promote abdominal fat, a feature of the Metabolic Syndrome. Effects of stress reduction interventions on abdominal fat are unknown. Forty-seven overweight/obese women (mean BMI = 31.2) were randomly assigned to a 4-month intervention or waitlist group to explore effects of a mindfulness program for stress eating. We assessed mindfulness, psychological distress, eating behavior, weight, cortisol awakening response (CAR), and abdominal fat (by dual-energy X-ray absorptiometry) pre- and posttreatment. Treatment participants improved in mindfulness, anxiety, and external-based eating compared to control participants. Groups did not differ on average CAR, weight, or abdominal fat over time. However, obese treatment participants showed significant reductions in CAR and maintained body weight, while obese control participants had stable CAR and gained weight. Improvements in mindfulness, chronic stress, and CAR were associated with reductions in abdominal fat. This proof of concept study suggests that mindfulness training shows promise for improving eating patterns and the CAR, which may reduce abdominal fat over time

Cell type-specific functions of Alzheimer's disease endocytic risk genes

Endocytosis is a key cellular pathway required for the internalization of cellular nutrients, lipids and receptor-bound cargoes. It is also critical for the recycling of cellular components, cellular trafficking and membrane dynamics. The endocytic pathway has been consistently implicated in Alzheimer's disease (AD) through repeated genome-wide association studies and the existence of rare coding mutations in endocytic genes. BIN1 and PICALM are two of the most significant late-onset AD risk genes after APOE and are both key to clathrin-mediated endocytic biology. Pathological studies also demonstrate that endocytic dysfunction is an early characteristic of late-onset AD, being seen in the prodromal phase of the disease. Different cell types of the brain have specific requirements of the endocytic pathway. Neurons require efficient recycling of synaptic vesicles and microglia use the specialized form of endocytosis—phagocytosis—for their normal function. Therefore, disease-associated changes in endocytic genes will have varied impacts across different cell types, which remains to be fully explored. Given the genetic and pathological evidence for endocytic dysfunction in AD, understanding how such changes and the related cell type-specific vulnerabilities impact normal cellular function and contribute to disease is vital and could present novel therapeutic opportunities. This article is part of a discussion meeting issue ‘Understanding the endo-lysosomal network in neurodegeneration’

The IgG1/G2 subclass shift – a sensitive, tissue non-specific marker for malignancy. Diagnostic performance with squamous cell carcinoma of the head and neck

A significant decrease in %IgG1 accompanied by an increase in %IgG2 in total serum IgG has been previously reported as a highly sensitive marker for detecting early stages of carcinomas of various localizations. Here we investigated the question as to whether this phenomenon is also observed in sera of patients with squamous cell carcinoma of the head–neck region (SCC-HN), and to evaluate its diagnostic performance in the post-operative monitoring. Using quantitative affinity chromatography, serum concentrations of IgG1, IgG2 and total IgG were determined in 81 patients with different stages of primary and untreated SCC-HN, in 51 SCC-HN patients in post-therapeutical follow up, and in 33 patients with organ matched benign diseases. The data were compared with a total of 174 healthy controls. It was found that (i) 105 SCC-HN patients exhibited a mean value of 56.0 ± 0.7% IgG1, which likewise differed from healthy controls (63.2 ± 0.5) and benign diseases (61.5 ± 1.0) with P < 0.0005, (ii) sensitivities and specificities for discriminating primary malignancies from healthy controls were 70 and 74% respectively, and from benign diseases 65 and 76%, (iii) highest sensitivities and specificities were observed with post-therapeutic cases suffering from tumour recurrence (88% and 75%) or patients with distant metastases (87% and 86%), (iv) apparently tumour-free post-therapeutic patients showed a mean %IgG1 not different from the normal value. The decrease in %IgG1 accompanied by increased %IgG2 is an efficient, sensitive and early marker of SCC-HN, which appears particularly useful for the post-therapeutic monitoring. © 1999 Cancer Research Campaig

Rhesus macaque personality, dominance, behavior, and health

Previous studies of nonhuman primates have found relationships between health and individual differences in personality, behavior, and social status. However, despite knowing these factors are intercorrelated, many studies focus only on a single measure, for example, rank. Consequently, it is difficult to determine the degree to which these individual differences are independently associated with health. The present study sought to untangle the associations between health and these individual differences in rhesus macaques (Macaca mulatta). We studied 85 socially housed macaques at the Oregon and California National Primate Research Centers, and used veterinary records to determine the number of injuries and illnesses for each macaque. We measured personality using 12 items from a well-established primate personality questionnaire, performed focal observations of behaviors, and calculated dominance status from directional supplant data. All twelve personality questionnaire items were reliable and were used to represent five of the six personality dimensions identified in rhesus macaques-Dominance, Confidence, Openness, Anxiety, and Friendliness (also known as Sociability). Following this, we fit generalized linear mixed effects models to understand how these factors were associated with an animal's history of injury and history of illness. In the models, age was an offset, facility was a random effect, and the five personality dimensions, behavior, sex, and dominance status were fixed effects. Number of injuries and illnesses were each best represented by a negative binomial distribution. For the injury models, including the effects did improve model fit. This model revealed that more confident and more anxious macaques experienced fewer injuries. For the illness models, including the fixed effects did not significantly improve model fit over a model without the fixed effects. Future studies may seek to assess mechanisms underlying these associations