Dissecting the Microscopic Steps of the Cyclophilin A Enzymatic Cycle on the Biological HIV-1 Capsid Substrate by NMR

Peptidyl-prolyl isomerases (PPIases) are emerging as key regulators of many diverse biological processes. Elucidating the role of PPIase activity in vivo has been challenging because mutagenesis of active-site residues not only reduces the catalytic activity of these enzymes but also dramatically affects substrate binding. Employing the cyclophilin A PPIase together with its biologically relevant and natively folded substrate, the N-terminal domain of the human immunodeficiency virus type 1 capsid (CA^N) protein, we demonstrate here how to dissect residue-specific contributions to PPIase catalysis versus substrate binding utilizing NMR spectroscopy. Surprisingly, a number of cyclophilin A active-site mutants previously assumed to be strongly diminished in activity toward biological substrates based only on a peptide assay catalyze the human immunodeficiency virus capsid with wild-type activity but with a change in the rate-limiting step of the enzymatic cycle. The results illustrate that a quantitative analysis of catalysis using the biological substrates is critical when interpreting the effects of PPIase mutations in biological assays.     

Exacerbated tissue destruction in DSS-induced acute colitis of OPN-null mice is associated with downregulation of TNF-alpha expression and non-programmed cell death

Osteopontin (OPN), a pro-inflammatory mediator, is constitutively expressed in normal gut and is upregulated in inflammatory colitis. To determine the significance of OPN in inflammatory bowel disease, we studied the development of acute, experimental colitis induced by dextran sulfate sodium (DSS) in OPN-null and wild-type (WT) mice. OPN expression was markedly increased in WT diseased colons, while a higher disease activity index, including spleen enlargement, bowel shortening, and mucosal destruction, was observed in OPN-null mice. Although peripheral blood neutrophil numbers were lower in DSS-treated OPN-null mice, tissue myeloperoxidase levels, reflecting enhanced neutrophil activity, were increased in the diseased colons. In comparison, lymphocyte numbers in peripheral blood were increased earlier than in DSS-treated WT mice. Despite a significantly greater spleen enlargement, flow cytometric analysis of splenocytes from the DSS-treated OPN-null mice revealed lower numbers of differentiated macrophages and (CD4+ and CD8alpha+) lymphocytes. Whereas pro-inflammatory cytokines, including G-CSF, RANTES, MIP1alpha, and TNF-alpha, were increased < 10-fold in DSS-treated WT splenocytes, expression of these cytokines was dramatically suppressed in the DSS-treated OPN-null splenocytes as well as gut tissues. The suppressed TNF-alpha response in OPN-null mice was reflected in a marked increase in non-apoptotic cell death in diseased colons. Collectively, these studies demonstrate that OPN is required for mucosal protection in acute inflammatory colitis.     

Prediction of the mechanical response of the femur with uncertain elastic properties

A mandatory requirement for any reliable prediction of the mechanical response of bones, based on quantitative computer tomography, is an accurate relationship between material properties (usually Young's modulus E) and bone density ρ. Many such E-ρ relationships are available based on different experiments on femur specimens with a large spread due to uncertainties. The first goal of this study is to pool and analyze the relevant available experimental data and develop a stochasticE-ρ relationship. This analysis highlights that there is no experimental data available to cover the entire density range of the human femur and that some "popular" E-ρ relationships are based on data that contains extreme scatter, while others are based on a very limited amount of information. The second goal is to use the newly developed stochastic E-ρ relationship in high-order finite element analyses (FEAs) for the computation of strains and displacements in two human proximal femurs, mimicking in vitro experiments. When compared with the experimental observations, the FEA predictions using the median of the stochastic E-ρ relationship follow the underlying distribution of the stochastic E-ρ relationship. Thus, most deviations of the FEA predictions from experimental observations can possibly be explained by uncertain elastic properties of the femur.     

Medieval History of the Duda’im Melon (<Emphasis Type="Italic">Cucumis melo</Emphasis>, Cucurbitaceae)

Medieval History of the Duda’im Melon ( Cucumis melo , Cucurbitaceae). Melons, Cucumis melo, are a highly polymorphic species for fruit characteristics. The melons that are the most valued are the ones that turn sweet when ripe, including the muskmelons, cantaloupes, and casabas. Others, including the elongate adzhur, conomon, and snake melons, are consumed when immature, like cucumbers. The duda’im melons, Cucumis melo Duda’im Group, are special, as their small, spherical, thin-fleshed, insipid but beautifully maroon, dark-orange, or brown-and-yellow striped ripe fruits are valued for ornament and especially for their lush fragrance. The distinctive properties of duda’im melons are matched with special names given to them in several languages and geographical areas, which have made possible tracing of the history of these melons to mid-9^th century Persia. From that region, duda’im melons diffused westward, likely facilitated by Islamic conquests, reaching North Africa and Andalusia in the 10^th century.     

Responsibility and vaccine nationalism in the Israeli-Palestinian conflict

In this article we articulate a case from moral responsibility to assist Palestinians living in the Occupied Palestinian Territory (OPT). We contextualize this responsibility by focusing on access to healthcare and the provision of vaccines against COVID-19. We specifically present two arguments from responsibility, one that is global or cosmopolitan, and one that is country-specific. For the latter, we focus on Israel.     

Daily and yearly burnout symptoms in Israeli shift work residents

Burnout is a syndrome of physical and emotional exhaustion that develops among individuals who are open to public demands. In view of their heavy work load and sleep deprivation, we decided to evaluate the impact of long working hours on burnout and psychological status among a sample of residents during the first 2 years of their residency. Seventy-eight residents participated in the study, all residents completed self-administered questionnaires, and their sleep-wake cycle was monitored by a wrist-worn actigraph for a period of 5-7 days. The questionnaires included a short form suitable for Experience Sampling Method (ESM), and a longer background Questionnaire. The results revealed that sleep duration, Work Load and the interaction between them, explain the Negative Mood the day after the night shift. However, positive mood, and fatigue were not affected by sleep duration or workload. In general, after one year of residency, residents become more stressed, less involved in the job, and had a high level of burnout and psychosomatic symptoms. However, after the second year, the burnout symptoms were almost the same as at the beginning except for the level of stress that remained high. Sleep duration was unrelated to the burnout symptoms.     

The substrate specificity of yeast hexokinase: reaction with D-arabinose oxime

By chromatography, electrophoresis, n.m.r. spectroscopy, and spectrophotometric assay, it has been shown that D-arabinose oxime acts as a weak substrate for yeast hexokinase. The enzyme-catalysed phosphorylation of the oxime, which exists as a mixture of E (80%) and Z (20%) acyclic forms in solution at equilibrium, is proposed to proceed via the transient formation of a furanoid species. Weak substrate-activity was also observed with 4-deoxy-D-xylo-hexose, but not with 5-deoxy-D-xylo-hexose. The relation of these and previous results concerning the carbohydrate-substrate specificity of yeast hexokinase in solution to X-ray crystallographic studies is discussed.     

Cyto-histological examination of post-vitellogenesis and final oocyte maturation in captive-reared striped bass

The ovarian development of captive-reared, striped bass Morone saxatilis was examined during a 10-week period encompassing the spawning season. Vitellogenic oocytes in March had a mean diameter of 838 ± 18 μm and did not grow significantly thereafter. Except from one non-hormone-treated fish, all females failed to undergo final oocyte maturation (FOM) and their ovaries became atretic with the onset of high spring temperatures. A clearing fixative was found useful in identifying early stages of atresia, evident by the absence of the germinal vesicle (GV). Final oocyte maturation of fish treated with gonadotropin-releasing hormone agonist (GnRHa) consisted of two phases. Early FOM lasted from 1 to 3 weeks, and was associated with lipid-droplet coalescence, and displacement of the GV and yolk globules to the peripheral cytoplasm. Late FOM lasted <24h, and consisted of yolk-globule coalescence and GV breakdown (GVBD). Ovulated eggs had completely coalesced lipid and yolk masses, with cortical alveoli lined against the cell wall. Both phases of FOM were associated with significant increases in oocyte diameter. Striped bass oocytes showed important morphological differences compared to oocytes of other members of the Moronidae family, in terms of percentage lipid content, chorion thickness and degree of hydration after ovulation.