Neglect in Aged Care - A Role for the Justice System?
The Royal Commission into Aged Care Quality and Safety's Interim Report entitled Neglect, identified multiple failings in meeting Australia's care needs of older people. Many of these have been examined from both health and regulation-based perspectives. However, although some of the issues uncovered relate to potential criminal acts, relatively few prosecutions of individuals for abuse in aged care contexts take place. In addition to the social invisibility of the aged in our community investigators may experience considerable difficulties in utilising traditional processes for criminal investigation and prosecution in matters involving aged care institutions.


Characteristics of Cellulose Nanofibrils from Transgenic Trees with Reduced Expression of Cellulose Synthase Interacting 1.
Cellulose nanofibrils can be derived from the native load-bearing cellulose microfibrils in wood. These microfibrils are synthesized by a cellulose synthase enzyme complex that resides in the plasma membrane of developing wood cells. It was previously shown that transgenic hybrid aspen trees with reduced expression of CSI1 have different wood mechanics and cellulose microfibril properties. We hypothesized that these changes in the native cellulose may affect the quality of the corresponding nanofibrils. To test this hypothesis, wood from wild-type and transgenic trees with reduced expression of CSI1 was subjected to oxidative nanofibril isolation. The transgenic wood-extracted nanofibrils exhibited a significantly lower suspension viscosity and estimated surface area than the wild-type nanofibrils. Furthermore, the nanofibril networks manufactured from the transgenics exhibited high stiffness, as well as reduced water uptake, tensile strength, strain-to-break, and degree of polymerization. Presumably, the difference in wood properties caused by the decreased expression of CSI1 resulted in nanofibrils with distinctive qualities. The observed changes in the physicochemical properties suggest that the differences were caused by changes in the apparent nanofibril aspect ratio and surface accessibility. This study demonstrates the possibility of influencing wood-derived nanofibril quality through the genetic engineering of trees.


Force history dependence of receptor-ligand dissociation.
Receptor-ligand bonds that mediate cell adhesion are often subjected to forces that regulate their dissociation via modulating off-rates. Off-rates control how long receptor-ligand bonds last and how much force they withstand. One should therefore be able to determine off-rates from either bond lifetime or unbinding force measurements. However, substantial discrepancies exist between the force dependence of off-rates derived from the two types of measurements even for the same interactions, e.g., selectins dissociating from their ligands, which mediate the tethering and rolling of leukocytes on vascular surfaces during inflammation and immune surveillance. We used atomic force microscopy to measure survival times of P-selectin dissociating from P-selectin glycoprotein ligand 1 or from an antibody in both bond lifetime and unbinding force experiments. By a new method of data analysis, we showed that the discrepancies resulted from the assumption that off-rates were functions of force only. The off-rates derived from forced dissociation data depended not only on force but also on the history of force application. This finding provides a new paradigm for understanding how force regulates receptor-ligand interactions.


Novel mouse model resistant to irreversible BTK inhibitors: a tool identifying new therapeutic targets and side effects.
Pharmacological inhibitors of Bruton tyrosine kinase (BTK) have revolutionized treatment of B-lymphocyte malignancies and show great promise for dampening autoimmunity. The predominant BTK inhibitors tether irreversibly by covalently binding to cysteine 481 in the BTK catalytic domain. Substitution of cysteine 481 for serine (C481S) is the most common mechanism for acquired drug resistance. We generated a novel C481S knock-in mouse model and, using a battery of tests, no overt B-lymphocyte phenotype was found. B lymphocytes from C481S animals were resistant to irreversible, but sensitive to reversible, BTK inhibitors. In contrast, irreversible inhibitors equally impaired T-lymphocyte activation in mice, mimicking the effect of treatment in patients. This demonstrates that T-lymphocyte blockage is independent of BTK. We suggest that the C481S knock-in mouse can serve as a useful tool for the study of BTK-independent effects of irreversible inhibitors, allowing for the identification of novel therapeutic targets and pinpointing potential side effects.


The interference of monoclonal antibodies with laboratory diagnosis: clinical and diagnostic implications.
Diagnostic test interference is due to the presence of material that falsely changes an analytic test result. The development of monoclonal antibodies is discussed with focus on their extensive use as both therapeutic and diagnostic agents. In this review the interference of monoclonal antibodies with laboratory test methods and the potential impact on clinical care is addressed. Recognition of the types of interference, endogenous and exogenous, and the varied mechanisms by which monoclonal antibodies may cause interference are discussed in this report. Review of the literature identifies cases which exemplify the issues facing laboratorians and clinicians and describe the impact on patients. Approaches to reducing and eliminating sources of interference are also addressed. Education of ordering clinicians concerning the possibility of interference in at-risk patients is key in limiting the impact on care. Laboratorians and medical practitioners should be cognizant of the risk of interference to avoid incorrect management of patients.


Respiratory syncytial virus infection in adults.
Respiratory syncytial virus (RSV), an enveloped RNA virus in the Paramyxovirus family, is the most important cause of lower respiratory tract infection in infants and young children, accounting for ~100,000 pediatric hospitalizations and 250 deaths annually in the United States. Despite primarily being recognized as a pediatric pathogen, RSV reinfection causes substantial disease in all adult populations, including healthy young persons, old and frail individuals, those with chronic obstructive pulmonary disease and immunocompromised patients. Most illnesses are mild in adults, but significant morbidity and mortality can develop. In contrast to infants, diagnosis of RSV infections is difficult due to low virus shedding, and optimal diagnosis requires molecular tests. Unfortunately, antiviral therapy is of limited benefit. Ribavirin and palivizumab are the only approved pharmacological agents for RSV treatment and prophylaxis, respectively, and are primarily used in infants; data regarding their usefulness in adults are limited. Currently, specific antiviral therapy is generally reserved for severely immunocompromised patients or severe respiratory failure. The greatest promise for reducing the impact of RSV in adults may be through immunization. However, an effective vaccine for RSV is not currently available.


Effect of chronic endogenous hypergastrinaemia on pancreatic growth and carcinogenesis in the hamster.
To examine the effect of gastrin on spontaneous and induced pancreatic carcinogenesis in the hamster. Two sets of experiments were carried out, one involving long term hypergastrinaemia and one involving cancer induction during hypergastrinaemia. The effect of hypergastrinaemia accomplished by gastric fundectomy was studied for eight months. Neither fundectomised hamsters nor sham operated controls developed premalignant or malignant pancreatic lesions. In the fundectomy group, the mean pancreatic weight, total protein content, and DNA content was increased by 28%, 25%, and 25% respectively. No such increases were found in fundectomised animals receiving a cholecystokinin-B receptor antagonist during the last 24 days of the experiment. In the cancer induction study, the effect of fundectomy on N-nitrosobis(2-oxopropyl) amine induced pancreatic carcinogenesis was studied for three months. There were no significant differences in the incidence or [3H]-thymidine labelling index of focal pancreatic lesions between fundectomised and sham operated control animals. Fundectomy with chronic hypergastrinaemia induces pancreatic hypertrophy, but does not enhance N-nitrosobis (2-oxopropyl)amine induced pancreatic carcinogenesis in the hamster. The increases in growth were inhibited by a cholecystokinin-B receptor antagonist, indicating that the trophic effect of fundectomy is mediated by gastrin.


Differential activation of the mouse beta-globin promoter by enhancers.
A series of plasmids was constructed to study the effect of two enhancers, the simian virus 40 72-base-pair repeat and the Harvey sarcoma virus 73-base-pair repeat, on the mouse beta maj-globin promoter. These plasmids contain the mouse beta maj-globin promoter linked to the Escherichia coli galK gene, thus allowing galactokinase enzyme activity to be used as a measure of promoter function. In CV-1 (primate) cells, it was found that an enhancer is required for optimal promoter activity and that the simian virus 40 (primate) enhancer increases galactokinase fourfold more than the Harvey sarcoma virus (mouse) enhancer. In L (mouse) cells, however, the Harvey sarcoma virus enhancer is 1.3-fold stronger than the simian virus 40 enhancer. These data support the hypothesis that enhancer activity can be species specific. Furthermore, when both enhancers are present on the same plasmid, their effect is additive on the beta-globin promoter whether the plasmid is in CV-1 cells or L cells.


Differential O- and Glycosphingolipid Glycosylation in Human Pancreatic Adenocarcinoma Cells With Opposite Morphology and Metastatic Behavior.
Changes in the glycosylation profile of cancer cells have been strongly associated with cancer progression. To increase our insights into the role of glycosylation in human pancreatic ductal adenocarcinoma (PDAC), we performed a study on O-glycans and glycosphingolipid (GSL) glycans of the PDAC cell lines Pa-Tu-8988T (PaTu-T) and Pa-Tu-8988S (PaTu-S). These cell lines are derived from the same patient, but show an almost opposite phenotype, morphology and capacity to metastasize, and may thus provide an attractive model to study the role of glycosylation in progression of PDAC. Gene-array analysis revealed that 24% of the glycosylation-related genes showed a ≥ 1.5-fold difference in expression level between the two cell lines. Subsequent validation of the data by porous graphitized carbon nano-liquid chromatography coupled to a tandem ion trap mass spectrometry and flow cytometry established major differences in O-glycans and GSL-glycans between the cell lines, including lower levels of T and sialylated Tn (sTn) antigens, neoexpression of globosides (Gb3 and Gb4), and higher levels of gangliosides in the mesenchymal-like PaTu-T cells compared to the epithelial-like PaTu-S. In addition, PaTu-S cells demonstrated a significantly higher binding of the immune-lectins macrophage galactose-type lectin and galectin-4 compared to PaTu-T. In summary, our data provide a comprehensive and differential glycan profile of two PDAC cell lines with disparate phenotypes and metastatic behavior. This will allow approaches to modulate and monitor the glycosylation of these PDAC cell lines, which opens up avenues to study the biology and metastatic behavior of PDAC.


Study of effectiveness of bioluminescent reporter phage assay on Y. pseudotuberculosis strains.
The method describes the phage-mediated transduction of a bioluminescent phenotype to cultivated Y. pseudotuberculosis cells which are subsequently measured using a microplate luminometer. Reporter phage assay is rapid detection technique and its efficiency is not affected by presence of contaminating bacteria, no sample preparation is needed and it has the ability to test multiple samples simultaneously in a 96-well microtiter plate format. Experiments were performed to develop the rapid detection technique for Y. pseudotuberculosis strains and study the ability of a reporter Yersinia phage to confer a bioluminescent signal to Y. pseudotuberculosis strains under different environmental conditions (media, temperature, bacterial number) for detection. Further, to determine if the Yersinia phage can detect Y. pseudotuberculosis in presence of other bacterial species. The results revealed that the developed reporter phage assay is not effective against wide range of Y. pseudotuberculosis. Y. pseudotuberculosis could be rapidly detected within 30 minutes at 28°C. The reporter phage assay could detect luminescence within 45 minutes when the bacterial cells were at the minimal concentration 105 cells/mL. The optimal detectable concentrations were 106-107 cells/mL at 28 and 37°C. The reporter phage assay could detect Y. pseudotuberculosis within 30 minutes in presence of other enteric bacteria without selective enrichment. It should be noted that the Yersinia reporter phage is specific to Yersinia pestis strains and it can be used to detect Y. pseudotuberculosis when samples exclude the existence of Y. pestis strains. In the presented study this aspect was foreseen.


Fluid overload at initiation of renal replacement therapy is associated with lack of renal recovery in patients with acute kidney injury.
Patients with acute kidney injury (AKI) requiring initiation of renal replacement therapy (RRT) have poor short- and long-term outcomes, including the development of dialysis dependence. Currently, little is known about what factors may predict renal recovery in this population. We conducted a single-center, retrospective analysis of 170 hospitalized adult patients with AKI attributed to acute tubular necrosis who required inpatient initiation of RRT. Data collection included patient characteristics, laboratory data, details of hospital course and degree of fluid overload at RRT initiation. The primary outcome was recovery of renal function to dialysis independence. Within 1 year of RRT initiation, 35.9% (61/170) of patients reached the primary end point of renal recovery. The median (interquartile range) duration of RRT was 11 (3-33) days and 83.6% (51/61) recovered prior to hospital discharge. Recovering patients had significantly less fluid overload at the time of RRT initiation compared to non-recovering patients (3.5 versus 9.3%, P = 0.004). In multivariate Cox proportional hazard regression analysis, a rise in percent fluid overload at dialysis initiation remained a significant negative predictor of renal recovery (hazard ratio 0.97, 95% confidence interval 0.95-1.00, P = 0.024). In patients with AKI, a higher degree of fluid overload at RRT initiation predicts worse renal recovery at 1 year. Clinical trials are needed to determine whether interventions targeting fluid overload may improve patient and renal outcomes.


Chronic acquired dyskeratotic papulosis of the face.
A Vietnam veteran who had been exposed to Agent Orange presented with a 28-year history of a papulonodular disease mainly affecting the face. Over those years, pin-sized papules had developed into dark nodules. Multiple biopsies of both fresh and mature papulonodules had been performed for routine histopathology and electron microscopy. Results had been positive for spongiosis with exocytosis, acanthosis, dyskeratotic keratinocytes, and marked incontinence of pigment, which together are suggestive of a previously unreported clinical entity that we term chronic acquired dyskeratotic papulosis.


DNA translocation to giant unilamellar vesicles during electroporation is independent of DNA size.
Delivery of naked DNA molecules into living cells via physical disruption of the membrane under electric pulses has potential biomedical applications ranging from gene electro-transfer, electro-chemotherapy, to gene therapy, yet the mechanisms involved in DNA transport remain vague. To investigate the mechanism of DNA translocation across the cell membrane, giant unilamellar vesicles (GUVs) were electroporated in the presence of DNA molecules keeping the size of the DNA molecules as a variable parameter. We experimentally determined the translocation efficiency for each size of the DNA molecule, to compare the results with the existing and conflicting theories of the translocation mechanism i.e. stochastic threading and bulk electrophoresis. We observed that the translocation efficiency is independent of DNA size (ranging from 25-20 000 bp, bp = base pairs), implying that DNA molecules translocate freely across the electro-pores in the lipid membrane in their native polymer conformation, as opposed to unravelling and threading through the electro-pore. Bulk electrophoretic mobility determines the relationship between translocation efficiency and the size of the DNA molecule. This research provides experimental evidence of the mechanistic understanding of DNA translocation across lipid membranes which is essential for devising efficient and predictable protocols for electric field mediated naked DNA delivery.


Atmospheric physics and Earth observations: mapping from space: the metric camera experiment.
The Spacelab metric camera experiment acquired stereoscopic high-resolution black-and-white and color infrared photographs of various regions of the world. In total, an area of about 11 million square kilometers was covered. Because of the delay in launching the shuttle until 28 November, illumination conditions were frequently poor over many candidate targets. However, unique high-quality images with a ground resolution of about 20 meters were obtained by increasing camera exposure time. Initial image analysis has shown that these images may be used for earth mapping at the scale 1:100,000.


Triggered ferroptotic polymer micelles for reversing multidrug resistance to chemotherapy.
Drug-tolerant persister cancer cells (PCCs) play an important role in the development of multidrug resistance (MDR) to anti-cancer drugs. This is due to the strong link between PCCs formation and epithelial-mesenchymal transition (EMT), as well as the low numbers of PCCs. In addition, PCC removal by traditional cytotoxic agents is poor due to the intrinsic high MDR activity in these cells. As a novel programmed cell death pathway, ferroptosis shows high potency to eliminate cells at the EMT state via manipulating intracellular redox homeostasis. The aim of this work was to utilize triggered ferroptotic polymer micelles for PCCs removal and MDR reversal both in vitro and in vivo. The micelles were made of arachidonic acid-conjugated amphiphilic copolymer that can enable rapid cargo release upon free radical-triggering in the tumor microenvironment. A potent ferroptotic inducer, RSL3 was encapsulated in the micelles to target the glutathione peroxidase 4 (GPX4). In the model resistant human ovarian adenocarcinoma cells, the RSL3 micelles were 30-fold more toxic than activatable control micelles due to the ferroptotic machinery. The lipid peroxidation-induced intracellular glutathione level reduction also made a contribution, which enhanced the potency of RSL3 for ferroptosis induction and enabled the drug-loaded micelles all-active. As an index of PCCs population, the level of CD133+ and aldehyde dehydrogenase (ALDH+) biomarker was significantly lower for the ferroptotic micelles in contrast to the control. The potency of ferroptotic micelles regarding PCCs reduction was proved by the in vitro soft agar colony forming assay. The in vivo anti-tumor efficacy of triggered micelles was further demonstrated in tumor-bearing nude mice in terms of PCCs biomarkers, tumor growth inhibition, mice survival, and GPX4 inhibition. This work demonstrates a novel strategy to overcome cancer MDR via the tailored ferroptotic micelles, which opens new avenues for managing resistant tumors.


Resource Use and Outcomes of Pediatric Congenital Heart Disease Admissions: 2003 to 2016.
Background Children with congenital heart disease (CHD) are known to consume a disproportionate share of resources, yet there are limited data concerning trends in resource use and mortality among admitted children with CHD. We hypothesize that charges in CHD-related admissions increased but that mortality improved over time. Methods and Results This study, including patients <18 years old with CHD, examined inpatient admissions from the nationally representative Kids' Inpatient Database from 2003 to 2016 in order to assess the frequency, medical complexity, and outcomes of CHD hospital admissions. A total of 859 843 admissions of children with CHD were identified. CHD admissions increased by 31.8% from 2003 to 2016, whereas overall pediatric admissions decreased by 13.4%. Compared with non-CHD admissions, those with CHD were more likely to be <1 year of age (80.5% versus 63.3%), and to have ≥1 complex chronic condition (39.7% versus 9.3%). For CHD admissions, mortality was higher (2.97% versus 0.31%) and adjusted median charges greater ($48 426 [interquartile range (IQR), $11.932-$161 048] versus $4697 [IQR, $2551-$12 301]) (P<0.0001 for all). Among CHD admissions, whereas adjusted median charges increased from $35 577 (IQR, $9303-$110 439) to $61 696 (IQR, $15 212-$219 237), mortality decreased from 3.2% to 2.7% (P for trend <0.0001). CHD admissions accounted for an increased proportion of all inpatient deaths, from 18.0% in 2003 to 24.5% in 2016. Conclusions Children admitted with CHD are 10 times more likely to die than those without CHD and have higher charges. Although the rate of mortality in CHD admissions decreased, children with CHD accounted for an increasing proportion of all pediatric inpatient deaths. Effective resource allocation is critical to optimize outcomes in these high-risk patients.


An Exosome-Rich Conditioned Medium from Human Amniotic Membrane Stem Cells Facilitates Wound Healing via Increased Reepithelization, Collagen Synthesis, and Angiogenesis.
Tissue regeneration is an essential requirement for wound healing and recovery of organs' function. It has been demonstrated that wound healing can be facilitated by activating paracrine signaling mediated by exosomes secreted from stem cells, since exosomes deliver many functional molecules including growth factors (GFs) and neurotrophic factors (NFs) effective for tissue regeneration. In this study, an exosome-rich conditioned medium (ERCM) was collected from human amniotic membrane stem cells (AMSCs) by cultivating the cells under a low oxygen tension (2% O2 and 5% CO2). The contents of GFs and NFs including keratinocyte growth factor, epidermal growth factor, fibroblast growth factor 1, transforming growth factor-β, and vascular endothelial growth factor responsible for skin regeneration were much higher (10-30 folds) in the ERCM than in normal conditioned medium (NCM). In was found that CM-DiI-labeled exosomes readily entered keratinocytes and fibroblasts, and that ERCM not only facilitated the proliferation of keratinocytes in normal condition, but also protected against H2O2 cytotoxicity. In cell-migration assay, the scratch wound in keratinocyte culture dish was rapidly closed by treatment with ERCM. Such wound-healing effects of ERCM were confirmed in a rat whole skin-excision model: i.e., the wound closure was significantly accelerated, remaining minimal crusts, by topical application of ERCM solution (4 × 109 exosome particles/100 μL) at 4-day intervals. In the wounded skin, the deposition of collagens was enhanced by treatment with ERCM, which was supported by the increased production of collagen-1 and collagen-3. In addition, enhanced angiogenesis in ERCM-treated wounds was confirmed by increased von Willebrand factor (vWF)-positive endothelial cells. The results indicate that ERCM from AMSCs with high concentrations of GFs and NFs improves wound healing through tissue regeneration not only by facilitating keratinocyte proliferation for skin repair, but also activating fibroblasts for extracellular matrix production, in addition to the regulation of angiogenesis and scar tissue formation.


Intra-Abdominal Heterotopic Cardiac Xenotransplantation: Pearls and Pitfalls.
Heterotopic cardiac xenotransplantation in the intra-abdominal position has been studied extensively in a pig-to-baboon model to define the optimal donor genetics and immunosuppressive regimen to prevent xenograft rejection. Extensive investigation using this model is a necessary stepping stone toward the development of a life-supporting animal model, with the ultimate goal of demonstrating suitability for clinical cardiac xenotransplantation trials. Aspects of surgical technique, pre- and post-operative care, graft monitoring, and minimization of infectious risk have all required refinement and optimization of heterotopic cardiac xenotransplantation over time. This review details non-immunologic obstacles relevant to this model described by our group and in the literature, as well as strategies that have been developed to address these specific challenges.


A high-density admixture map for disease gene discovery in african americans.
Admixture mapping (also known as "mapping by admixture linkage disequilibrium," or MALD) provides a way of localizing genes that cause disease, in admixed ethnic groups such as African Americans, with approximately 100 times fewer markers than are required for whole-genome haplotype scans. However, it has not been possible to perform powerful scans with admixture mapping because the method requires a dense map of validated markers known to have large frequency differences between Europeans and Africans. To create such a map, we screened through databases containing approximately 450000 single-nucleotide polymorphisms (SNPs) for which frequencies had been estimated in African and European population samples. We experimentally confirmed the frequencies of the most promising SNPs in a multiethnic panel of unrelated samples and identified 3011 as a MALD map (1.2 cM average spacing). We estimate that this map is approximately 70% informative in differentiating African versus European origins of chromosomal segments. This map provides a practical and powerful tool, which is freely available without restriction, for screening for disease genes in African American patient cohorts. The map is especially appropriate for those diseases that differ in incidence between the parental African and European populations.


Diet influences the functions of the human intestinal microbiome.
Gut microbes programme their metabolism to suit intestinal conditions and convert dietary components into a panel of small molecules that ultimately affect host physiology. To unveil what is behind the effects of key dietary components on microbial functions and the way they modulate host-microbe interaction, we used for the first time a multi-omic approach that goes behind the mere gut phylogenetic composition and provides an overall picture of the functional repertoire in 27 fecal samples from omnivorous, vegan and vegetarian volunteers. Based on our data, vegan and vegetarian diets were associated to the highest abundance of microbial genes/proteins responsible for cell motility, carbohydrate- and protein-hydrolyzing enzymes, transport systems and the synthesis of essential amino acids and vitamins. A positive correlation was observed when intake of fiber and the relative fecal abundance of flagellin were compared. Microbial cells and flagellin extracted from fecal samples of 61 healthy donors modulated the viability of the human (HT29) colon carcinoma cells and the host response through the stimulation of the expression of Toll-like receptor 5, lectin RegIIIα and three interleukins (IL-8, IL-22 and IL-23). Our findings concretize a further and relevant milestone on how the diet may prevent/mitigate disease risk.


Treatment of urodelans based on temperature dependent infection dynamics of Batrachochytrium salamandrivorans.
The recently emerged chytrid fungus Batrachochytrium salamandrivorans currently causes amphibian population declines. We hypothesized that temperature dictates infection dynamics of B. salamandrivorans, and that therefore heat treatment may be applied to clear animals from infection. We examined the impact of environmental temperature on B. salamandrivorans infection and disease dynamics in fire salamanders (Salamandra salamandra). Colonization of salamanders by B. salamandrivorans occurred at 15°C and 20°C but not at 25°C, with a significantly faster buildup of infection load and associated earlier mortality at 15°C. Exposing B. salamandrivorans infected salamanders to 25°C for 10 days resulted in complete clearance of infection and clinically cured all experimentally infected animals. This treatment protocol was validated in naturally infected wild fire salamanders. In conclusion, we show that B. salamandrivorans infection and disease dynamics are significantly dictated by environmental temperature, and that heat treatment is a viable option for clearing B. salamandrivorans infections.


Neuroprotective Role of L-NG-Nitroarginine Methyl Ester (L-NAME) against Chronic Hypobaric Hypoxia with Crowding Stress (CHC) Induced Depression-Like Behaviour.
Improper neuroimmune responses following chronic stress exposure have been reported to cause neuronal dysfunctions leading to memory impairment, anxiety and depression like behaviours. Though several factors affecting microglial activation and consequent alteration in neuro-inflammatory responses have been well studied, role of NO and its association with microglia in stress induced depression model is yet to be explored. In the present study, we validated combination of chronic hypobaric hypoxia and crowding (CHC) as a stress model for depression and investigated the role of chronic stress induced elevated nitric oxide (NO) level in microglia activation and its effect on neuro-inflammatory responses in brain. Further, we evaluated the ameliorative effect of L-NG-Nitroarginine Methyl Ester (L-NAME) to reverse the stress induced depressive mood state. Four groups of male Sprague Dawley rat were taken and divided into control and CHC stress exposed group with and without treatment of L-NAME. Depression like behaviour and anhedonia in rats were assessed by Forced Swim Test (FST) and Sucrose Preference Test (SPT). Microglial activation was evaluated using Iba-1 immunohistochemistry and proinflammatory cytokines were assessed in the hippocampal region. Our result showed that exposure to CHC stress increased the number of active microglia with corresponding increase in inflammatory cytokines and altered behavioural responses. The inhibition of NO synthesis by L-NAME during CHC exposure decreased the number of active microglia in hippocampus as evident from decreased Iba-1 positive cells. Further, L-NAME administration decreased pro-inflammatory cytokines in hippocampus and improved behaviour of rats. Our study demonstrate that stress induced elevation of NO plays pivotal role in altered microglial activation and consequent neurodegenerative processes leading to depression like behaviour in rat.


Actin induction during PMA and cAMP-dependent signal pathway activation in Entamoeba histolytica trophozoites.
Activation of PKC or cAMP-dependent signalling pathways in Entamoeba histolytica triggers the phosphorylation of proteins involved in actin rearrangements necessary for adhesion and locomotion. Analogous motifs to SRE and CRE sequences--known to respond to PMA and cAMP--were identified within the 5' regulatory region (5'RR) of one of the parasite actin genes. These sequences could be involved in the actin transcriptional upregulation reported during signalling. To test this hypothesis, a plasmid containing the 5'RR of the actin gene fused to the bacterial neomycin gene (neo) was used for stable transfection. Expression of neo and endogenous actin was measured after stimulation of transfected amoebae by PMA and dcAMP. It was found that both compounds induced neo and actin expression and showed a co-operative effect in the induction of neo. Induction by PMA or dcAMP failed if the directing amoebic 5'RR lacked SRE and CRE motifs. Transfection of amoebae with plasmid constructs, containing either progressive deletions of the actin 5'RR or site-directed mutations of the SRE and CRE-like motifs, corroborated that these sequences and a co-ordinated participation of PKC- and PKA-activated transcription factors are responsible for the increments in neo and actin mRNAs. In vivo, these PMA and cAMP-response elements could play an important role in regulating actin expression and organization in signalling processes activated during tissue invasion.


Mass shootings, firearm injuries, and mental health.
The correlation between mass shootings, firearm injuries, and mental health is an ongoing polarized debate within the U.S., making it essential to develop public policy on mental illness and firearm injuries exacerbated by a significant increase in firearm sales in March 2020. Although many mass shooters are labeled "mentally ill," mental illness is only present in a small minority of cases. Most mentally ill people are never violent but are more likely to be the victims of violence. Easy access to firearms and a triggering event deriving from social and economic inequalities are primary causes of mass shootings and growing online radicalization. Radicalization can easily lead to fatal firearm injuries, particularly for individuals with diagnosed or undiagnosed mental illness. Proposed solutions include permits for firearms purchase, including a 25-year-old age limit, universal background checks, and banning large-capacity magazines. Additionally, a speedy and effective law enforcement response is the sole factor and the most reliable way to stop a mass shooting once it has started. The research identified several other recommendations, including expanding Medicare and mental health care access, expanding school safety and law enforcement training, and promoting public education about mental health and firearm safety.


Screening for congenital heart diseases by murmurs using telemedical phonocardiography.
A large proportion of congenital heart diseases (CHD) remain undetected during pregnancy or even after birth. Many of them generate turbulent blood flow, resulting heart murmur. Doppler ultrasound cardiotocography (CTG) is suitable for the assessment of the fetal heart rate and some derived parameters, but it is inadequate for detecting heart murmurs. Although comprehensive examination can be carried out with echocardiography, it is expensive and requires expertise; therefore, it is not applicable for widespread screening. This paper presents a new possibility for screening for some CHDs using phonocardiography, which can be combined with Doppler ultrasound CTG as an extension of it. Furthermore it can be carried out at home allowing repeated measurements, which increases also the reliability of filtering out innocent murmurs. The diagnostic capability of this screening method is supported by a large number of evaluated fetal heart sound records. Moreover, according to experiences pregnant women prefer this reliable, easy to use method, which facilitates their examination.


Comparison of childhood thyroid cancer prevalence among 3 areas based on external radiation dose after the Fukushima Daiichi nuclear power plant accident: The Fukushima health management survey.
The 2011 Great East Japan Earthquake led to a subsequent nuclear accident at the Fukushima Daiichi Nuclear Power Plant. In its wake, we sought to examine the association between external radiation dose and thyroid cancer in Fukushima Prefecture. We applied a cross-sectional study design with 300,476 participants aged 18 years and younger who underwent thyroid examinations between October 2011 and June 2015. Areas within Fukushima Prefecture were divided into three groups based on individual external doses (≥1% of 5 mSv, <99% of 1 mSv/y, and the other). The odds ratios (ORs) and 95% confidence intervals of thyroid cancer for all areas, with the lowest dose area as reference, were calculated using logistic regression models adjusted for age and sex. Furthermore, the ORs of thyroid cancer for individual external doses of 1 mSv or more and 2 mSv or more, with the external dose less than 1 mSv as reference, were calculated. Prevalence of thyroid cancer for the location groups were 48/100,000 for the highest dose area, 36/100,000 for the middle dose area, and 41/100,000 for the lowest dose area. Compared with the lowest dose area, age-, and sex-adjusted ORs (95% confidence intervals) for the highest-dose and middle-dose areas were 1.49 (0.36-6.23) and 1.00 (0.67-1.50), respectively. The duration between accident and thyroid examination was not associated with thyroid cancer prevalence. There were no significant associations between individual external doses and prevalence of thyroid cancer. External radiation dose was not associated with thyroid cancer prevalence among Fukushima children within the first 4 years after the nuclear accident.


Approaches used to prevent and reduce the use of restrictive practices on adults with learning disabilities: Protocol for a realist review.
The use of restrictive practices has significant adverse effects on the individual, care providers and organisations. This review will describe how, why, for whom, and in what circumstances approaches used by healthcare organisations work to prevent and reduce the use of restrictive practices on adults with learning disabilities. Evidence from the literature will be synthesised using a realist review approach - an interpretative, theory-driven approach to understand how complex healthcare approaches work in reducing the use of restrictive practices in these settings. In step 1, existing theories will be located to explore what approaches work by consulting with key topic experts, holding consultation workshops with healthcare professionals, academics, and experts by experience, and performing an informal search to help develop an initial programme theory. A systematic search will be performed in the second step in electronic databases. Further searches will be performed iteratively to test particular subcomponents of the initial programme theory, which will also include the use of the CLUSTER approach. Evidence judged as relevant and rigorous will be used to test the initial programme theory. In step three, data will be extracted and coded inductively and deductively. The final step will involve using a realist logic of analysis to refine the initial programme theory in light of evidence. This will then provide a basis to describe and explain what key approaches work, why, how and in what circumstances in preventing and reducing the use of restrictive practices in adults with learning disabilities in healthcare settings. Findings will be used to provide recommendations for practice and policymaking. In accordance with the guidelines, this realist review protocol was registered with the International Prospective Register of Systematic Reviews (PROSPERO) on 4th December 2019 (CRD42019158432).


Research strategies for capturing transactional models of development: the limits of the possible.
Transactional models have informed research design and interpretation in studies relevant to developmental psychopathology. Bidirectional effects between individuals and social contexts have been found in many behavioral and cognitive domains. This review will highlight representative studies where the transactional model has been explicitly or implicitly tested. These studies include experimental, quasiexperimental, and naturalistic designs. Extensions of the transactional model have been made to interventions designed to target different aspects of a bidirectional system in efforts to improve developmental outcomes. Problems remain in the need to theoretically specify structural models and to combine analyses of transactions in the parent-child relationship with transactions in the broader social contexts. Longitudinal studies with sufficient time points to assess reciprocal processes continue to be important. Such longitudinal investigations will permit identifying developmental periods where the child or the context may be most influential or most open to change.


Cellular and Extracellular Proteome of the Animal Pathogen Corynebacterium silvaticum, a Close Relative of Zoonotic Corynebacterium ulcerans and Corynebacterium pseudotuberculosis.
Corynebacterium silvaticum is a newly described animal pathogen, closely related to the emerging human pathogen Corynebacterium ulcerans and Corynebacterium pseudotuberculosis, a major pathogen of small ruminants. In this study, proteins of a whole cell and a shaving fraction and the exoproteome of C. silvaticum strain W25 were analyzed as a first proteome study of this species. In total, 1305 proteins were identified out of 2013 proteins encoded by the W25 genome sequence and number of putative virulence factors were detected already under standard growth conditions including phospholipase D and sialidase. An up to now uncharacterized trypsin-like protease is by far the most secreted protein in this species, indicating a putative role in pathogenicity. Furthermore, the proteome analyses carried out in this study support the recently published taxonomical delineation of C. silvaticum from the closely related zoonotic Corynebacterium species.


Transcranial sonography in spinocerebellar ataxia type 2.
To study the use of transcranial brain parenchyma sonography (TCS), in particular the echogenic signal in the substantia nigra (SN), in patients with spinocerebellar ataxia type 2 (SCA2), recently recognized as an uncommon cause of parkinsonism. Six consecutive and unrelated SCA2 patients without parkinsonian signs, 30 consecutive patients with Parkinson's disease (PD), and 30 healthy, age- and sexmatched controls were prospectively studied with TCS according to a standardized protocol. Four (67 %) of the six SCA2 patients exhibited SN hyperechogenicity. In two patients, the hyperechogenicity was classified as moderate (unilateral in both) and in two as marked. Differences between the SN echogenicity of the SCA2 group or the PD group and controls were statistically significant (p < 0.001), while there was no difference between the two groups of patients. Transcranial brain parenchyma sonography detects SN hyperechogenicity in the majority of patients with SCA2 without parkinsonian signs. It would be important to reproduce our TCS findings in a larger number of SCA2 patients, as well as to test their possible significance in differentiating SCA2 from other types of SCA.


The Impact of Expanding Telehealth-Delivered Dietary Interventions on Long-Term Cardiometabolic Health.
A healthy diet is an important protective factor to prevent cardiometabolic disease. Traditional face-to-face dietary interventions are often episodic, expensive, and may have limited effectiveness, particularly among older adults and people living in rural areas. Telehealth-delivered dietary interventions have proven to be a low-cost and effective alternative approach to improve dietary behaviors among adults with chronic health conditions. In this study, we developed a validated agent-based model of cardiometabolic health conditions to project the impact of expanding telehealth-delivered dietary interventions among older adults in the state of Georgia, a state with a large rural population. We projected the incidence of major cardiometabolic health conditions (type 2 diabetes, hypertension, and high cholesterol) with the implementation of telehealth-delivered dietary interventions versus no intervention among all older adults and 3 subpopulations (older adults with diabetes, hypertension, and high cholesterol, separately). The results showed that expanding telehealth-delivered dietary interventions could avert 22,774 (95% confidence interval [CI]: 22,091-23,457) cases of type 2 diabetes, 19,732 (19,145-20,329) cases of hypertension, and 18,219 (17,672-18,766) cases of high cholesterol for 5 years among older adults in Georgia. The intervention would have a similar effect in preventing cardiometabolic health conditions among the 3 selected subpopulations. Therefore, expanding telehealth-delivered dietary interventions could substantially reduce the burden of cardiometabolic health conditions in the long term among older adults and those with chronic health conditions.


Production and Characterization of High Value Prebiotics From Biorefinery-Relevant Feedstocks.
Hemicellulose, a structural polysaccharide and often underutilized co-product stream of biorefineries, could be used to produce prebiotic ingredients with novel functionalities. Since hot water pre-extraction is a cost-effective strategy for integrated biorefineries to partially fractionate hemicellulose and improve feedstock quality and performance for downstream operations, the approach was applied to process switchgrass (SG), hybrid poplar (HP), and southern pine (SP) biomass at 160°C for 60 min. As a result, different hemicellulose-rich fractions were generated and the chemical characterization studies showed that they were composed of 76-91% of glucan, xylan, galactan, arabinan, and mannan oligosaccharides. The hot water extracts also contained minor concentrations of monomeric sugars (≤18%), phenolic components (≤1%), and other degradation products (≤3%), but were tested for probiotic activity without any purification. When subjected to batch fermentations by individual cultures of Lactobacillus casei, Bifidobacterium bifidum, and Bacteroides fragilis, the hemicellulosic hydrolysates elicited varied responses. SG hydrolysates induced the highest cell count in L. casei at 8.6 log10 cells/ml, whereas the highest cell counts for B. fragilis and B. bifidum were obtained with southern pine (5.8 log10 cells/ml) and HP hydrolysates (6.4 log10 cells/ml), respectively. The observed differences were attributed to the preferential consumption of mannooligosaccharides in SP hydrolysates by B. fragilis. Lactobacillus casei preferentially consumed xylooligosaccharides in the switchgrass and southern pine hydrolysates, whereas B. bifidum consumed galactose in the hybrid poplar hydrolysates. Thus, this study (1) reveals the potential to produce prebiotic ingredients from biorefinery-relevant lignocellulosic biomass, and (2) demonstrates how the chemical composition of hemicellulose-derived sources could regulate the viability and selective proliferation of probiotic microorganisms.


Respectful death: a model for end-of-life care.
The Respectful Death Model (RDM) is a research-based, holistic, and practical model developed to improve end-of-life care. A respectful death is one which supports dying patients, their families, and professionals in the completion of life cycles and can be used by all members of the healthcare team. The model is a process method commencing with the establishment of a therapeutic relationship with the dying patient and his or her family and, as a result, their stories are heard and incorporated into the care plan. This article demonstrates that hospice and palliative care nurses have been practicing this model since the origination of care of the dying. Other topics addressed are the current culture toward death in the United States, the roles of nurses in the RDM, and the barriers and benefits of the RDM. Recommendations for future research in end-of-life care also are addressed.


[Effect of aluminum poisoning and gamma-irradiation on the antioxidant system of the body and study of possibilities for correction].
The influence of selenium and rutin on the antioxidant protection system of the organisms in connection with Al-intoxication and gamma-irradiation was investigated. It has been determined that selenium increases the antioxidant protection caused by aluminium. Under simultaneous action of aluminium and gamma-irradiation upon the organism selenium stimulated accumulation of toxic lipids peroxide oxidation products. Rutin action in the situations under investigation caused weakening of antioxidant protection.


Emerging Roles of the Human Solute Carrier 22 Family.
The human Solute Carrier 22 family (SLC22), also termed the organic ion transporter family, consists of 28 distinct multi-membrane spanning proteins, which phylogenetically cluster together according to their charge specificity for organic cations (OCTs), organic anions (OATs) and organic zwitterion/cations (OCTNs). Some SLC22 family members are well characterized in terms of their substrates, transport mechanisms and expression patterns, as well as their roles in human physiology and pharmacology, whereas others remain orphans with no known ligands. Pharmacologically, SLC22 family members play major roles as determinants of the absorption and disposition of many prescription drugs, and several including the renal transporters, OCT2, OAT1 and OAT3 are targets for many clinically important drug-drug interactions. In addition, mutations in some of these transporters (SLC22A5 (OCTN2) and SLC22A12 (URAT1) lead to rare monogenic disorders. Genetic polymorphisms in SLC22 transporters have been associated with common human disease, drug response and various phenotypic traits. Three members in this family were deorphaned in very recently: SLC22A14, SLC22A15 and SLC22A24, and found to transport specific compounds such as riboflavin (SLC22A14), anti-oxidant zwitterions (SLC22A15) and steroid conjugates (SLC22A24). Their physiologic and pharmacological roles need further investigation. This review aims to summarize the substrates, expression patterns and transporter mechanisms of individual SLC22 family members and their roles in human disease and drug disposition and response. Gaps in our understanding of SLC22 family members are described. Significance Statement In recent years, three members of the SLC22 family of transporters have been deorphaned and found to play important roles in the transport of diverse solutes. New research has furthered our understanding of the mechanisms, pharmacological roles, and clinical impact of SLC22 transporters. This minireview provides overview of SLC22 family members of their physiologic and pharmacologic roles, the impact of genetic variants in the SLC22 family on disease and drug response, and summary of recent studies deorphaning SLC22 family members.


The economics of employing a counsellor in general practice: analysis of data from a randomised controlled trial.
Counselling is currently adopted in many general practices, despite limited evidence of clinical and cost effectiveness. To compare direct and indirect costs of counsellors and general practitioners (GPs) in providing care to people with emotional problems. We carried out a prospective, randomized controlled trial of non-directive counselling and routine general practice care in 14 general practices in north London. Counsellors adhered to a Rogerian model of counselling. The counselling sessions ranged from one to 12 sessions over 12 weeks. As reported elsewhere, there were no differences in clinical outcomes between the two groups. Therefore, we conducted a cost minimisation analysis. We present only the economic outcomes in this paper. Main outcome measures were cost data (service utilisation, travel, and work absence) at baseline, three months, and nine months. One hundred and thirty-six patients with emotional problems, mainly depression, took part. Seventy patients were randomised to the counsellors and 66 to the GPs. The average direct and indirect costs for the counsellor was 162.09 Pounds more per patient after three months compared with costs for the GP group; however, over the following six months the counsellor group was 87.00 Pounds less per patient than the GP group. Over the total nine-month period, the counsellor group remained more expensive per patient. Referral to counselling is no more clinically effective or expensive than GP care over a nine-month period in terms of direct plus indirect costs. However, further research is needed to establish indirect costs of introducing a counsellor into general practice.


"I can't bend it and it hurts like mad": direct observation of gout consultations in routine primary health care.
Gout is the most common form of inflammatory arthritis and is associated with considerable co-morbidity. It is usually managed in the primary care setting with a combination of lifestyle modification and pharmacological therapy. This study describes patterns of communication about gout observed in interactions between patients and primary care practitioners during routine consultations. Secondary analysis of video-recordings of individual healthcare consultations between patients and a range of primary care practitioners (including general practitioners, practice nurses, podiatrists and dietitians) from an archived database. Consultations that included any discussion about gout were eligible for inclusion (n = 31) and were not restricted to those where gout was the main presenting complaint. The consultation transcripts were analysed using a qualitative inductive approach from clinical and linguistic perspectives and supplemented with visual observation of the interactions. Two main themes emerged from the data; the importance of gout and 'telling' versus 'listening' in consultations. The first theme had two distinct strands; gout as an incidental part of the consultation and the impact of gout on patients. A trend towards more didactic practitioner communication encompassed by the second theme occurred at many different consultation points including diagnosis, in more general post-diagnosis discussion, and when discussing biochemical test results and lifestyle advice. In contrast, when discussion about treatment with medicines occurred a tendency towards a greater degree of listening to patients was observed. Our observation of the communication patterns in these consultations illustrates an inherent complexity of gout consultations in primary care. Gout may be more important to patients than is often apparent to practitioners in routine consultations. Consultation management needs to take into account the impact of the condition and the balance of information provided around lifestyle advice versus long-term management with medicines.


Selenium contamination of the Grasslands, a major California waterfowl area.
In a recent study at Kesterson Reservoir in California, selenium was shown to cause mortality and deformities in embryos of aquatic birds. The present study was conducted to determine if selenium or other contaminants in agricultural drainwater used for marsh management were likely to cause similar adverse effects in the nearby Grasslands area. Selenium concentrations were elevated (greater than 15 ppm, dry-weight) in livers of some birds of all species collected from the Grasslands. Mean selenium concentrations in all species sampled in the South Grasslands were significantly higher (P less than 0.05) than those from the "control site", the Volta Wildlife Area. Mean selenium levels in black-necked stilts (Himantopus mexicanus) from the South Grasslands (35.6 ppm) were similar (P greater than 0.05) to levels in stilts from Kesterson (46.4 ppm), but means for American avocets (Recurvirostra americana) from the South Grasslands (67.3 ppm) were higher (P less than 0.05) than those from Kesterson (28.4 ppm). Bird eggs and fish from the Grasslands also contained elevated levels of selenium. Concentrations of eight heavy metals in fish generally reflected those patterns previously found in water entering the study areas. Of the organochlorines detected in fish, only DDE occurred at concentrations potentially harmful to birds (6.1 and 3.0 ppm, wet weight, at two South Grassland sites). The effect on avian health or reproduction of the other contaminants, singly or in combination, could not be determined. However, selenium levels were apparently sufficiently elevated in 1984 to have caused adverse effects on avian reproduction in the South Grasslands.


Prevalence and pathology of lungworm infection in bottlenose dolphins Tursiops truncatus from southwest Florida.
Parasitism of the respiratory system is a relatively common finding in stranded cetaceans; however, no systematic investigations regarding the severity, distribution, and clinical consequences of these infections in bottlenose dolphins Tursiops truncatus have been conducted previously. The present study determined the prevalence of lungworm infections in dead stranded (n=22) and live bottlenose dolphins (n=44) from southwestern Florida, USA, during the period from 2003 to 2005. Dead stranded bottlenose dolphins were necropsied and lungs were examined visually, by palpation, and histologically for lesions consistent with verminous pneumonia. When present, nematodes were counted, measured, and identified to species based upon their morphology. Dolphin feces and blowhole swabs were collected and examined for nematode larvae. Lungworm prevalence was 77% in dead animals (n=22). The lesions in most cases were mild, chronic, and not the primary cause of death. Only 13% of dead animals examined had patent infections, with larvae present in blowhole and fecal cytology, and only 18% of animals had intact worms present at necropsy, with a geometric mean intensity of infection of 22.6 worms animal(-1). Intact worms were identified as either Halocercus lagenorhynchi or Skrjabinalius cryptocephalus. The highest prevalence of active infections was found in neonates and calves, including 1 stillborn calf. For free-ranging animals, all blowhole swabs (n=44) were negative, and fecal cytology (n=22) showed a 3% prevalence of patent infection. Findings from the present study support the theory that bottlenose dolphins can be infected transplacentally by lungworms. The impact that such infections may have on neonatal survival is unknown; however, these infections could increase neonatal mortality.


Crystal structures of Trypanosoma brucei oligopeptidase B broaden the paradigm of catalytic regulation in prolyl oligopeptidase family enzymes.
Oligopeptidase B cleaves after basic amino acids in peptides up to 30 residues. As a virulence factor in bacteria and trypanosomatid pathogens that is absent in higher eukaryotes, this is a promising drug target. Here we present ligand-free open state and inhibitor-bound closed state crystal structures of oligopeptidase B from Trypanosoma brucei, the causative agent of African sleeping sickness. These (and related) structures show the importance of structural dynamics, governed by a fine enthalpic and entropic balance, in substrate size selectivity and catalysis. Peptides over 30 residues cannot fit the enzyme cavity, preventing the complete domain closure required for a key propeller Asp/Glu to fix the catalytic His and Arg in the catalytically competent conformation. This size exclusion mechanism protects larger peptides and proteins from degradation. Similar bacterial prolyl endopeptidase and archael acylaminoacyl peptidase structures demonstrate this mechanism is conserved among oligopeptidase family enzymes across all three domains of life.


Influence of Na+ and anions on the dimorphic transition of Candida albicans.
The effect of Na+ (Cl- or gluconate salt) on growth and dimorphic potential of the pathogenic yeast Candida albicans has been examined. Profiles of germ tube formation as a function of salt addition, pH and temperature indicated Na+ inhibition of germ tube outgrowth at high ambient pH (pH 8.0) which was exacerbated by replacement of Cl- with gluconate (as an impermeant analogue). At acidic pH (pH 5.5) and permissive temperature (37 degrees C), gluconate alone promoted the dimorphic transition. Rates of glucose-induced medium acidification and plasma membrane H(+)-ATPase activity have been measured to assess whether salt treatments could retard the cytoplasmic alkalinization known to precede germ tube formation. The precise site of Na+ action remains unclear but the anion effects may be interpreted in terms of anion-exchanger and channel activity acting to modulate cytosolic pH.


Strangers at the benchside: research ethics consultation.
Institutional ethics consultation services for biomedical scientists have begun to proliferate, especially for clinical researchers. We discuss several models of ethics consultation and describe a team-based approach used at Stanford University in the context of these models. As research ethics consultation services expand, there are many unresolved questions that need to be addressed, including what the scope, composition, and purpose of such services should be, whether core competencies for consultants can and should be defined, and how conflicts of interest should be mitigated. We make preliminary recommendations for the structure and process of research ethics consultation, based on our initial experiences in a pilot program.


Stereodivergent Silylzincation of α-Heteroatom-Substituted Alkynes.
Zinc reagents (Me2PhSi)2Zn and [(Me3Si)3Si]2Zn undergo highly regio- and stereoselective addition across the carbon-carbon triple bond of nitrogen-, sulfur-, oxygen-, and phosphorus-substituted terminal alkynes in the absence of copper or any other catalyst. Both reagents yield exclusively β-isomers, and the stereoselectivity is determined by the silyl group: Me2PhSi for cis or (Me3Si)3Si for trans. These stereodivergent silylzincation protocols offer an efficient access to heteroatom-substituted vinylsilanes with either double bond geometry, including trisubstituted vinylsilanes by one-pot electrophilic substitution of the intermediate C(sp(2))-Zn bond by copper(I)-mediated carbon-carbon bond formation.


The protein C pathway in inflammatory bowel disease: the missing link between inflammation and coagulation.
Traditionally described as a major anti-coagulant system, the protein C (PC) pathway, consisting of thrombomodulin, the endothelial cell protein C receptor and activated PC (APC), is gaining increasing attention as an important regulator of microvascular inflammation. Although they possess several anti-inflammatory and cytoprotective functions, the expression and function of the components of the PC pathway is downregulated during inflammation. Recent evidence suggests that the PC pathway is defective in patients with inflammatory bowel disease (IBD) and that restoring its function has anti-inflammatory effects on cultured intestinal microvascular endothelial cells and in animal models of colitis. Here, we propose that the PC pathway has an important role in governing intestinal microvascular inflammation and might provide a novel therapeutic target in the management of IBD.


[Epidemiological and evolutionary aspects of patients aged 65 and over operated on for fracture of the upper end of the femur: prospective observational study].
Fractures of the upper end of the femur (FUEF) are a serious complication for the elderly, affecting 1,3 million patients a year worldwide (1). Studies dedicated to the post-operative follow-up of patients operated on for hip fracture in the long term are rare. The aim was to determine the epidemiological and evolutionary profile focusing on morbidity and mortality (survival and functional and cognitive decline) in the year following surgery for FUEF in patients aged over 65. Methods: Prospective observational study. Geriatric data were collected at the time of surgical management of FUEF with postoperative assessment of vital, functional and cognitive status. Results: Over a period of 24 months, 119 patients underwent surgery in the Orthopaedics-Traumatology Department of the University Hospital of Annaba. The patients were initially all independent (Instrumental Activities of Daily Living [IADL] 4 ≥ 3) and had no pre-operative cognitive disorders. There were 32 men and 87 women. The mean age was (77,4 ± 8,2) years with extremes of 65 and 100 years. All lesions were unilateral, with a predominance on the right. Most were trochanteric fractures (72 %). The most common aetiological circumstances were falls following domestic accidents. Eighty-seven patients received spinal anaesthesia, 19 received a general anaesthetic and 13 received an epidural. The average intervention time was 5,11 ± 4,17 days, with extremes of zero and 18 days. Depending on the treatment performed, 14 Dynamic hip Screw (DHS), three centro-medullary nailings, 33 plate blades and 24 plate nails and 15 gamma nails, 28 Moore's prosthesis, 2 screwings. The average length of stay was seven days. The one-year mortality rate for the 119 patients was 19,3 %. Patients who died after one year were malnourished (13,2 %, p = 0,05) with high comorbidity (ASA 2(78 %) and ASA 3(13 %) (p < 0,01 and p = 0,05). Of the survivors, 27 % had postoperative cognitive decline. Death occurred on average at 82,17 days. Conclusion: FUEF remains a real public health problem in terms of its prevalence, prognosis and cost. The mortality rate of patients aged over 65 years at one year post-operatively following an ESF was 19 %, which calls for the creation of a specialised orthogeriatric unit.


Emergent exploratory thoracotomy with military casualties: contemporary prehospital management and outcome (see editorial page 619).
Chest trauma is among the most common types of trauma, corresponding to 10% of trauma patients admitted to hospitals. In the military setting, thoracic trauma was reported as a significant cause of death. With well-timed treatment, chest trauma is regarded as survivable. Emergency thoracotomy (ET) is considered when the patient with trauma to the chest needs immediate resuscitation. Survival rate is reported as low as 1% in some reports and 20% in others. The survival rate depends on injury mechanism, protocols for intervention, and other decompressive procedures. To determine parameters that may impact survival of ET. We conducted a retrospective cohort study to compare prehospital and in-hospital data regarding ET in the emergency department (ED) versus the operating room (OR). Between 2009 and 2017, 6532 casualties presented to the ED; 1125 with trauma to the chest. Fifty-four of those with chest trauma underwent ET in the hospital (4.8%), 22 (41%) in the ED, and 32 (59%) in the OR. The overall mortality of the ET subgroup was 48%. With regard to thoracotomies, 19/22 of patients (86%) who underwent ET in the ED died compared to 2/28 in the OR (13%). Utilizing ET after chest trauma with appropriate clinical indications, well-trained personnel, and prompt transportation poses a significant challenge, but may be associated with better survival than that reported previously with military casualties. Adoption of indications and timed allocation to the OR may improve outcomes with chest trauma casualties.


Reaction of NO(2) with selected conjugated alkenes.
The gas phase reactions of selected alkenes (isoprene, myrcene, ocimene, and 1,3-cyclohexadiene) with NO2 under dark condition have been investigated at T ∼ 298 K and P ∼ 760 Torr of purified air. The kinetic studies were performed under pseudo-first-order conditions using a large excess of NO2 concentration to those of the alkenes. The rate coefficients (in 10(-19) cm(3) molecule(-1) s(-1)) obtained are 1.1 ± 0.2 for isoprene, 2.5 ± 0.3 for myrcene, 8.5 ± 1.2 for ocimene, and 15 ± 1 for 1,3-cyclohexadiene. Several products were identified by using in situ Fourier transform infrared (FT-IR) spectrometry, and acetone was found to be the major product from the reactions of NO2 with myrcene and ocimene, with a formation yield of 22 ± 3% and 26 ± 7%, respectively. The oxidation products from the reactions of NO2 with isoprene and 1,3-cyclohexadiene were found to be mainly nitro compounds identified by FT-IR spectroscopy. Reaction mechanisms were proposed to account for the products observed.


Characterization of the Aspergillus ochraceoroseus aflatoxin/sterigmatocystin biosynthetic gene cluster.
Production of carcinogenic aflatoxins has been reported from members of Aspergillus section Flavi, Aspergillus section Nidulantes and a newly proposed Aspergillus section Ochraceorosei that consists of Aspergillus ochraceoroseus and A. rambellii. Unlike members of section Flavi, A. ochraceoroseus and A. rambellii have been shown to accumulate both aflatoxin (AF) and the aflatoxin precursor sterigmatocystin (ST). Alhough morphologically distinct from A. nidulans, molecular characterization of A. ochraceoroseus AF/ST genes and physiological characteristics of AF/ST production indicated that A. ochraceoroseus is more closely related to A. nidulans than to A. flavus. Knowing that the A. nidulans ST gene cluster is organized differently from the A. flavus AF gene cluster, we determined the genetic organization of the AF/ST biosynthetic cluster in A. ochraceoroseus. Sequencing of overlapping lambda clones and genomic PCR fragments obtained by gene-walking techniques demonstrated that the A. ochraceoroseus AF/ST gene cluster is organized much like the A. nidulans ST gene cluster except that the region from aflN to aflW is located directly upstream of aflC and in reverse orientation such that aflW represents the distal end and aflY the proximal end of the cluster. The A. ochraceoroseus cluster genes demonstrated 62-76% nucleotide identity to their A. nidulans ST cluster gene homologs. Transformation of an A. nidulans aflR mutant with the A. ochraceoroseus aflR restored ST production in A. nidulans transformants. PCR amplification of A. rambellii genomic DNA demonstrated that the AF/ST gene cluster is organized in the same manner as that of A. ochraceoroseus.


Therapeutic development in amyotrophic lateral sclerosis.
Amyotrophic lateral sclerosis (ALS) is the most common motor neuron disease in adults. It is almost invariably lethal within a few years after the onset of symptoms. No effective treatment is currently available beyond supportive care and riluzole, a putative glutamate release blocker linked to modestly prolonged survival. This review provides a general overview of preclinical and clinical advances during recent years and summarizes the literature regarding emerging therapeutic approaches, focusing on their molecular targets. A systematic literature review of PubMed was performed, identifying key clinical trials involving molecular therapies for ALS. In addition, the ALS Therapy Development Institute website was carefully analyzed, and a selection of ALS clinical trials registered at ClinicalTrials.gov has been included. In the last several years, strategies have been developed to understand both the genetic and molecular mechanisms of ALS. Several therapeutic targets have been actively pursued, including kinases, inflammation inhibitors, silencing of key genes, and modulation or replacement of specific cell populations. The majority of ongoing clinical trials are investigating the safety profiles and tolerability of pharmacologic, gene, and cellular therapies, and have begun to assess their effects on ALS progression. Currently, no therapeutic effort seems to be efficient, but recent findings in ALS could help accelerate the discovery of an effective treatment for this disease.


Fertility-conserving surgery for young women with stage IA1 cervical cancer: safety and access.
To estimate the safety of fertility-conserving surgery for stage IA1 cervical cancer and to analyze predictors of access to conization. We analyzed women with stage IA1 cervical cancer aged 40 years or younger who were diagnosed between 1988 and 2005 and recorded in the Surveillance, Epidemiology, and End Results database. The outcomes of hysterectomy were compared with fertility-conserving conization. Clinical and demographic characteristics were compared using chi2. Multivariable logistic regression models were constructed to examine predictors of conization. Survival was examined using multivariable Cox proportional hazards models and the Kaplan-Meier method. A total of 1,409 patients were identified, including 841 (60%) who underwent hysterectomy and 568 (40%) who underwent conization. In a multivariable logistic regression of factors associated with conization, Asian patients, single women, those diagnosed in the later years of the study, and those residing in the eastern United States were more likely to have fertility-conserving surgery. Compared with women younger than 30 years, those older than 35 years were 78% (odds ratio 0.22, 95% confidence interval [CI] 0.16-0.30) less likely to undergo conization. In a Cox proportional hazards model accounting for other prognostic variables, there was no difference in survival (hazard ratio 0.65, 95% CI 0.23-1.47) between conization and hysterectomy. Five-year survival for women who underwent conization was 98% (95% CI 96-99%), compared with 99% (95% CI 97-99%) for those treated with hysterectomy. Fertility-conserving surgery is safe for young women with stage IA1 squamous cell carcinoma of the cervix. Young women with microinvasive cervical tumors should weigh the risks and benefits of conization in the context of individual preferences and tumor characteristics.


Breast reconstruction after failed conservation.
We report our experience of using latissimus dorsi myocutaneous flaps after failed conservation for breast carcinoma. Twenty-nine patients were treated by two methods of reconstruction. Seventeen patients with central recurrent tumours and three patients with radiation necrosis of the breast were treated by total mastectomy and latissimus dorsi reconstitution with silicone implant. Nine patients underwent latissimus dorsi reconstruction with preservation of the nipple for recurrent peripheral tumours. After a mean follow up period of 20.2 months no local recurrences have been observed but a longer period of follow-up is necessary to evaluate the likely long term recurrence rate.


NMR and EPR studies of the bis(pyridine) and bis(tert-butyl isocyanide) complexes of iron(III) octaethylchlorin.
The NMR and EPR spectra of a series of pyridine complexes [(OEC)Fe(L)2]+ (L = 4-Me2NPy, Py, and 4-CNPy) have been investigated. The EPR spectra at 4.2 K suggest that, with a decrease of the donor strength of the axial ligands, the complexes change their ground state from (d(xy))2 (d(xz)d(yz))3 to (d(xz)d(yz))4 (d(xy))1. The NMR data from 303 to 183 K show that at any temperature within this range the chemical shifts of pyrrole-8,17-CH2 protons increase with a decrease in the donor strength of the axial ligands. The full peak assignments of the [(OEC)Fe(L)2]+ complexes of this study have been made from COSY and NOE difference experiments. The pyrrole-8,17-CH2 and pyrroline protons show large chemical shifts (hence indicating large pi spin density on the adjacent carbons which are part of the pi system), while pyrrole-12,13-CH2 and -7,18-CH2 protons show much smaller chemical shifts, as predicted by the spin densities obtained from molecular orbital calculations, both Hückel and DFT; the DFT calculations additionally show close energy spacing of the highest five filled orbitals (of the Fe(II) complex) and strong mixing of metal and chlorin character in these orbitals that is sensitive to the donor strength of the axial substituents. The pattern of chemical shifts of the pyrrole-CH2 protons of [(OEC)Fe(t-BuNC)2]+ looks somewhat like that of [(OEC)Fe(4-Me2NPy)2]+, while the chemical shifts of the meso-protons are qualitatively similar to those of [(OEP)Fe(t-BuNC)2]+. The temperature dependence of the chemical shifts of [(OEC)Fe(t-BuNC)2]+ shows that it has a mixed (d(xz)d(yz))4 (d(xy))1 and (d(xy))2 (d(xz),d(yz))3 electron configuration that cannot be resolved by temperature-dependent fitting of the proton chemical shifts, with a S = 5/2 excited state that lies somewhat more than 2kT at room temperature above the ground state; the observed pattern of chemical shifts is the approximate average of those expected for the two S = 1/2 electronic configurations, which involve the a-symmetry SOMO of a planar chlorin ring with the unpaired electron predominantly in the d(yz) orbital and the b-symmetry SOMO of a ruffled chlorin ring with the unpaired electron predominantly in the d(xy) orbital. A rapid interconversion between the two, with calculated vibrational frequency of 22 cm(-1), explains the observed pattern of chemical shifts, while a favoring of the ruffled conformation explains the negative chemical shift (and thus the negative spin density at the alpha-pyrroline ring carbons), of the pyrroline-H of [TPCFe(t-BuNC)2]CF3SO3 (Simonneaux, G.; Kobeissi, M. J. Chem. Soc., Dalton Trans. 2001, 1587-1592). Peak assignments for high-spin (OEC)FeCl have been made by saturation transfer techniques that depend on chemical exchange between this complex and its bis-4-Me2NPy adduct. The contact shifts of the pyrrole-CH2 and meso protons of the high-spin complex depend on both sigma and pi spin delocalization due to contributions from three of the occupied frontier orbitals of the chlorin ring.


The Need to Routinely Convert Emergency Cricothyroidotomy to Tracheostomy: A Systematic Review and Meta-Analysis.
Traditional surgical teaching advocates converting emergency cricothyroidotomies to tracheostomies to mitigate the risk of subglottic stenosis. A conversion procedure that may risk losing a tenuous airway should have clear benefits over risks. We aimed to evaluate the necessity of routine cricothyroidotomy to tracheostomy conversion by conducting a systematic review and meta-analysis of contemporary literature. We performed a systematic review of experimental and observational studies (published between January 1, 2008, and March 1, 2021) reporting hospital outcomes of adults aged ≥18 years who underwent emergency cricothyroidotomies or tracheostomies. We followed PRISMA guidelines and assessed quality of data using GRADE methodology. Meta-analysis pooled incidence of procedure-specific complications (bleeding, subglottic stenosis, and others) using Freeman-Tukey double arcsine transformation and sensitivity analysis addressed survival bias. A total of 18 studies including 1246 patients were analyzed. Incidence of bleeding (5 [1 to 11]% vs 3 [1 to 7]%), subglottic stenosis (0 [0 to 3]% vs 0 [0 to 0]%) and other complications (12 [8 to 16]% vs 13 [5 to 23]%) were similar among patients undergoing emergency cricothyroidotomy or tracheostomy. Sensitivity analysis evaluating the incidence of complications among only survivors found similar results. Only one study reported complications attributable to cricothyroidotomy to tracheostomy conversion. Subglottic stenosis, the main harm conversion seeks to avoid, appears to be a rare complication after cricothyroidotomy. We did not find evidence supporting routine need to convert cricothyroidotomies to tracheostomies; for many patients, conversion is unlikely to rectify complications attributable to emergency cricothyroidotomy. However, our findings cannot be generalized to patients who require prolonged or permanent airway cannulation. Providers should consider performing cricothyroidotomy to tracheostomy selectively when the benefits clearly outweigh the risks of disrupting a secured airway.


Fuzzy service control of queueing systems.
We consider queueing systems in which the service rate is the controlled variable. The cost depends on the queue length and selected rate. The objective is to choose the service rate dynamically, based on the state of the system so as to minimize the average cost over an infinite horizon. Six classes, either known in the literature or new, are studied in detail: queueing systems with vacations, single-server queueing systems with and without switching costs, and tandem queueing systems with and without service costs. A novel approach is presented here using fuzzy control to solve these problems. Simulation shows that the approach is efficient and promising, especially in cases where analytical solutions do not exist.


Factors leading to osteoarthritis after eccentric rotational acetabular osteotomy.
We performed eccentric rotational acetabular osteotomy consecutively in 273 hips in 248 patients with hip dysplasia. Twenty-one patients were male and 227 were female. The average age was 37.7 years at the time of the index operation. Twenty-two hips had no osteoarthritis, 129 had early osteoarthritis, 117 had advanced osteoarthritis, and five had end-stage osteoarthritis. Twenty-six hips also were treated with concomitant intertrochanteric valgus osteotomy. Patients were followed for a minimum of 5 years after surgery. The average Harris hip score improved from 71 points preoperatively to 92 points at the final followup. Nine hips were converted to total hip arthroplasty as a result of deterioration after surgery. Kaplan-Meier survivorship analysis 15 years after the index operation indicated 97% in pre- and early stages and 87% in advanced and end stages when the end point was total hip arthroplasty. Risk factors identified by multivariate analysis for development of osteoarthritis of the hip 5 years after the index operation were body mass index (24 kg/m2 or more), concomitant valgus osteotomy, operative year between 1989 and 1992, a postoperative center-edge angle of Wiberg less than 25 degrees , and a postoperative horizontal distance of the femoral head from the tear drop (40 mm or more). Level IV, therapeutic study. See the Guidelines for Authors for a complete description of levels of evidence.


Phlebectasia of the external jugular vein associated with duplication of the internal jugular vein.
The most common cause of a neck mass that increases in size on the Valsalva maneuver is laryngocele. Jugular phlebectasia is a congenital dilation of the jugular vein, which is extremely rare in adults, may present similarly. Duplication of the internal jugular vein (IJV), which is usually encountered coincidentally, is another rare anatomic variation. Isolated cases or cases associated with IJV ectasia have been reported. We report on an adult patient with coexisting external jugular phlebectasia and internal jugular duplication on the same side.


Effects of cDNA hybridization on translation of encephalomyocarditis virus RNA.
Cell-free translation of the RNA of encephalomyocarditis virus was examined after hybridization of chemically synthesized cDNA fragments to different sites of the 5' noncoding region of the viral RNA. The following results were obtained. The binding of cDNA fragments to the first 41 nucleotides, to the poly(C) tract (between nucleotides 149 and 263), and to the sequence between nucleotides 309 and 338 did not affect translation of the viral RNA; the binding of cDNA fragments to the sequence between nucleotides 420 and 449 caused a slight inhibition; and the binding of fragments to eight different sites between nucleotides 450 and the initiator AUG codon (nucleotide 834) caused high degrees of inhibition. The results suggest that the first part of the 5' untranslated region, at least to nucleotide 338, may not be required for encephalomyocarditis viral RNA translation; however, the region near nucleotide 450 is important for translation of the viral RNA. The possibility that initiation occurs at an internal site is discussed.


[Surgeries on the carotid arteries].
The article lays emphasis on the main causes of restenosis after endarterectomy from the bifurcation of the carotid artery and shows the methods of its prevention. A new method for endarterectomy of the bifurcation of the carotid artery is suggested, which consists in reimplantation of the internal carotid artery. The authors claim this method to be optimal in combined affection: atherosclerotic constriction of carotid artery bifurcation and pathological tortuosity of the internal carotid artery. The authors possess an experience in 351 operations on the carotid arteries, 177 of them were conducted for loops and kinks of the carotid arteries; 152, for endarterectomy in atherosclerotic affection of the carotid arteries; and 22 for other, less frequently encountered affections of the carotid arteries.


Coronary artery bypass graft patency: assessment with true ast imaging with steady-state precession versus gadolinium-enhanced MR angiography.
To compare the accuracy of multisection true fast imaging with steady-state precession (FISP) with gadolinium-enhanced magnetic resonance (MR) angiography for the detection of coronary artery bypass graft patency. Twenty-five patients with coronary artery bypass grafts who had recently undergone conventional coronary angiography underwent MR angiography with a 1.5-T system. True FISP angiographic images were acquired in transverse and coronal planes. Coronal cardiac-gated MR angiography was performed with 0.2 mL per kilogram of body weight of gadopentetate dimeglumine injected at a rate of 2 mL/sec. With conventional angiography as the reference standard, the sensitivity, specificity, and accuracy of each technique for the detection of graft patency were determined. Image quality and duration of analysis were determined by two experienced radiologists. In 25 patients, 46 of 56 venous grafts were patent and 22 of 23 arterial grafts were patent. In all grafts at true FISP angiography, sensitivity for patency was 84% (57 of 68 grafts), specificity was 45% (five of 11 grafts), and accuracy was 78% (62 of 79 grafts). At MR angiography, sensitivity was 85% (58 of 68 grafts), specificity was 73% (eight of 11 grafts), and accuracy was 84% (66 of 79 grafts) (difference not significant). Image quality scores were similar with both techniques, but duration of analysis was significantly longer with MR angiography than with true FISP angiography (29 minutes 24 seconds vs 14 minutes 6 seconds, P <.001). Accuracy for detection of coronary artery bypass graft patency was similar with gadolinium-enhanced MR angiography and true FISP angiography, with a trend toward more false-positive findings for occlusion and reduced visualization of arterial grafts with true FISP angiography.


Diverse interactions between the individual mutations in a double mutant at the active site of staphylococcal nuclease.
In principle, the quantitative effect of a second mutation on a mutant enzyme may be antagonistic, absent, partially additive, additive, or synergistic with respect to the first mutation. Depending on the kinetic or thermodynamic parameter measured, the D21E and R87G mutations of staphylococcal nuclease exhibit four of these five categories of interaction in the double mutant. While Vmax of the R87G single mutant of staphylococcal nuclease is 10(4.8)-fold lower than that of the wild-type enzyme and the Vmax of the D21E single mutant is 10(3.0)-fold below that of wild type, the double mutant D21E + R87G was found to lose a factor of only 10(4.1) in Vmax relative to wild type, rather than the product of the two single mutations (10(7.8)). These results suggest antagonistic structural effects of the individual R87G and D21E mutations. An alternative explanation for the nonadditivity of effects, namely, the separate functioning of these residues in a stepwise mechanism involving the prior attack of water on phosphorus followed by protonation of the leaving group by Arg-87, is unlikely since no enzyme-bound phosphorane intermediate (less than 1% of [enzyme]) was found under steady-state conditions on the R87G mutant by 31P NMR at 242.9 MHz. Like the effects on Vmax, quantitatively similar antagonistic effects of the two mutations were detected on the binding of divalent cations in binary enzyme-Ca2+ and enzyme-Mn2+ complexes and in the ternary enzyme-Ca2(+)-5'-pdTdA complex, suggesting that the effects on Vmax result from antagonistic structural changes at the Ca2+ binding site. Simple additive weakening effects of the two mutations were found on the binding of the substrate 5'-pdTdA, in both the absence and the presence of the divalent cations, Mn2+ and Ca2+. However, synergistic effects of the two mutations were found on the binding of the substrate analogue 3',5'-pdTp, profoundly weakening its binding to the double mutant in both the absence and the presence of divalent cations. Such synergistic effects of the two mutations may result from negative cooperativity or strain in the binding of 3',5'-pdTp to the wild-type enzyme. It is concluded that the quantitative interactions of two active-site mutations of an enzyme can vary greatly depending on which parameter of the enzyme is measured. When the two mutations interact in the same way on several parameters, a common underlying mechanism is suggested.


[Metabolism of 25-hydroxyvitamin D3 and its regulation in rats during hypokinesis].
The influence of short-(7 days) and long-term (28 days) hypokinesia on 25-hydroxyvitamin D3 metabolism was investigated in rats fed on a normal calcium (0.6%), normal phosphorus (0.6%), vitamin D-supplemented diet. The animals were given a single intraperitoneal dose of tritiated [26,27-3H]25(OH)D3 (200 pmol) eighteen hours before sacrifice. [3H]Labelled vitamin D3 metabolites were separated by high performance liquid chromatographic procedure, and their radioactivity levels in serum, kidney, intestinal mucosa and femoral bone were measured. Long-term hypokinesia resulted in decreased levels of [3H]1.25(OH)2D3 and increased levels of [3H]24.25(OH)2D3 in serum and kidney (3.15 +/- 0.62 vs. 4.33 +/- 0.41% and 5.34 +/- 0.69 vs. 3.76 +/- 0.29% for [3H]1.25(OH)2D3 and [3H]24.25(OH)2D3 in serum; 7.52 +/- 0.69 vs. 11.6 +/- 0.79% and 9.33 +/- 0.55 vs. 5.94 +/- 0.24% for those in kidney). The levels of [3H]1.25(OH)2D3 as well as of [3H] 24.25(OH)2D3 were decreased in intestinal mucosa and bone (21.5 +/- 1.46 vs. 30.1 +/- 3.04% and 7.30 +/- 0.58 vs. 9.18 +/- 0.78% for [3H]1.25(OH)2D3 and [3H]24.25(OH)2D3 in intestinal mucosa; 6.39 +/- 06.5 vs. 11.5 +/- 1.64% and 7.78 +/- 0.71 vs. 13.9 +/- 1.28% for those in bone). The data obtained suggest a suppressed synthesis of 1.25(OH)2D3 and enhanced production of 24.25(OH)2D3 in kidney as well as a diminished binding of 24.25(OH)2D3 in intestinal mucosa and bone in hypothetic rats. Possible causes of variations in biosynthesis of vitamin D3 active metabolites, and role of these variations in the disorders of calcium metabolism and bone state during hypokinesia are discussed.


Probing Chemical Complexity of Amyloid Plaques in Alzheimer's Disease Mice using Hyperspectral Raman Imaging.
One of the distinctive pathological features of Alzheimer's disease (AD) is the deposition of amyloid plaques within the brain of affected individuals. These plaques have traditionally been investigated using labeling techniques such as immunohistochemical imaging. However, the use of labeling can disrupt the structural integrity of the molecules being analyzed. Hence, it is imperative to employ label-free imaging methods for noninvasive examination of amyloid deposits in their native form, thereby providing more relevant information pertaining to AD. This study presents compelling evidence that label-free and nondestructive confocal Raman imaging is a highly effective approach for the identification and chemical characterization of amyloid plaques within cortical regions of an arcAβ mouse model of AD. Furthermore, this investigation elucidates how the spatial correlation of Raman signals can be exploited to identify robust Raman marker bands and discern proteins and lipids from amyloid plaques. Finally, this study uncovers the existence of distinct types of amyloid plaques in the arcAβ mouse brain, exhibiting significant disparities in terms of not only shape and size but also molecular composition.


Dexmedetomidine Infusion Versus Placebo During Light or Deep Anesthesia on Postoperative Delirium in Older Patients Undergoing Major Noncardiac Surgery: A Pilot Randomized Factorial Trial.
Postoperative delirium (POD) is common among older surgical patients and may be affected by dexmedetomidine and depth of anesthesia. We designed this pilot study to assess the feasibility of comparing dexmedetomidine with normal saline during light versus deep anesthesia on POD in older patients undergoing major noncardiac surgery. In this pilot randomized factorial study, 80 patients aged 60 years or older undergoing major noncardiac surgery were randomized (1:1:1:1) to receive dexmedetomidine infusion 0.5 μg/kg/h or normal saline placebo during light (bispectral index [BIS] target 55) or deep (BIS target 40) anesthesia. Feasibility end points included consent rate and dropout rate, timely enrollment, blinded study drug administration throughout surgery, no inadvertent unmasking, achieving BIS target throughout >70% of surgery duration, and the process of twice-daily POD screening. In addition, we estimated the POD incidences in the 2 control groups (placebo and deep anesthesia) and treatment effects of dexmedetomidine and light anesthesia. Between November 1, 2021, and June 30, 2022, 78 patients completed the trial (mean [standard deviation, SD] age, 69.6 [4.6] years; 48 male patients [62%]; dexmedetomidine-deep, n = 19; dexmedetomidine-light, n = 20; placebo-deep, n = 19; placebo-light, n = 20). This study had a high consent rate (86%) and a low dropout rate (2.5%). Average recruitment was 5 patients at each center per month. Dexmedetomidine and normal saline were administered in a blinded fashion in all patients. Unmasking did not occur in either group. Approximately 99% of patients received the scheduled study drug infusion throughout the surgery. Approximately 81% of patients achieved the BIS targets throughout >70% of the surgery duration. The scheduled twice-daily POD screening was completed without exception. Overall, 10 of the 78 patients (13%; 95% confidence interval [CI], 7%-22%) developed POD. For the 2 reference groups, POD was observed in 7 of the 39 patients (17.9%; 95% CI, 9%-32.7%) in the placebo group and 7 of the 38 patients (18.4%; 95% CI, 9.2%-33.4%) in the deep anesthesia group. Regarding the treatment effects on POD, the estimated between-group difference was -10% (95% CI, -28% to 7%) for dexmedetomidine versus placebo, and -11% (95% CI, -28% to 6%) for light versus deep anesthesia. The findings of this pilot study demonstrate the feasibility of assessing dexmedetomidine versus placebo during light versus deep anesthesia on POD among older patients undergoing major noncardiac surgery, and justify a multicenter randomized factorial trial.


Efficient production of functional mRNA mediated by RNA polymerase I in Trypanosoma brucei.
The unicellular eukaryote Trypanosoma brucei evades the immune defence of its mammalian host by antigenic variation. The genes for variant-specific surface glycoproteins (VSGs) are expressed within large multicistronic transcription units. Mature messenger RNAs are produced by trans-splicing and polyadenylation. A remarkable feature of the transcription of VSG genes is its insensitivity to the RNA polymerase II inhibitor alpha-amanitin. This has led to the speculation that RNA polymerase I, normally only involved in the transcription of ribosomal RNA genes, also mediates expression of these surface antigen genes. In higher eukaryotes, however, transcripts produced by RNA polymerase I were found to be poor substrates for processing into mature mRNAs. In contrast, we show here that the RNA polymerase I of T. brucei can mediate the efficient production of functional mRNA for neomycin phosphotransferase. This exceptional ability may be related to the unusual way in which pre-mRNAs are capped in trypanosomes. In most eukaryotes, mRNAs are modified at their 5' end by a capping activity associated with RNA polymerase II; in trypanosomes, mRNAs acquire their 5'-cap from capped mini-exon donor RNA by trans-splicing, a process that could be independent of the RNA polymerase producing the pre-mRNA.


Mentalization and relationships with parents as predictors of eating disordered behavior.
Our study postulated that deficient mentalization abilities and maladaptive relationships with parents may be risk factors in the genesis of eating disorder (ED). The study examined 34 female inpatients with anorexia nervosa-binging/purging type and 35 matched non-ED controls. Data indicated that anorexia nervosa-binging/purging type patients presented significantly lower mentalization levels and reported significantly lower quality of current relationships with their parents, compared with non-ED controls. The combination of the mentalization and relationship quality variables showed an additive effect for some ED-related symptoms but not others. Moreover, high mentalization reduced ED symptomatology even if relationship quality with parents was deficient. Findings supported the role of low mentalization as a risk factor, and of high mentalization as a protective factor, in the formation of ED.


Temporal and spatial characteristics of highly pathogenic avian influenza outbreaks in China during 2004 to 2015.
Identifying the temporal and spatial characteristics of highly pathogenic avian influenza (HPAI) outbreaks is very important for developing effective and appropriate countermeasures against HPAI and promoting sustainable development in the poultry industry. This study aimed to analyze four aspects of the temporal and spatial characteristics of HPAI outbreaks in China, including the frequency of HPAI outbreaks, numbers of dead animals (died or culled), types of HPAI viruses, and species of infected animals. Temporal characteristics showed that the frequency of HPAI outbreaks decreased and then increased, with some years deviating from the main trend in 2004 to 2010 and 2011 to 2015, while the largest number of dead animals due to HPAI outbreaks was in 2005. During 2004 to 2015, HPAI H5N1 was the major type of HPAI virus, and chickens had the greatest risk of being infected with HPAI, followed by ducks and geese. The HPAI outbreaks had obvious seasonal effects clustered in January to February, June, and November. Spatial characteristics revealed that outbreaks were more frequent in Xinjiang, Hubei, and Guangdong but caused a larger number of dead animals in Liaoning and Shanxi. HPAI H5N1 appeared in 25 provinces, while HPAI H5N2 was mainly localized in Hebei and Jiangsu, and HPAI H5N6 occurred in Heilongjiang, Jiangsu, Hunan, and Guangdong. HPAI viruses were most frequently detected in chickens and wild birds in northern China, while the majority of HPAI infections were identified in chickens, ducks, and geese in southern China. Regionally, HPAI outbreaks were most frequent in the western region but resulted in larger number of animals dying or being culled in the eastern region. These findings could provide a new understanding of the distributional characteristics of HPAI outbreaks and offer prospects for better prevention and control strategies.