A role for PACE4 in osteoarthritis pain: evidence from human genetic association and null mutant phenotype.
The aim of this study was to assess if genetic variation in the PACE4 (paired amino acid converting enzyme 4) gene Pcsk6 influences the risk for symptomatic knee osteoarthritis (OA). Ten PCSK6 single nucleotide polymorphisms were tested for association in a discovery cohort of radiographic knee OA (n=156 asymptomatic and 600 symptomatic cases). Meta-analysis of the minor allele at rs900414 was performed in three additional independent cohorts (total n=674 asymptomatic and 2068 symptomatic). Pcsk6 knockout mice and wild-type C57BL/6 mice were compared in a battery of algesiometric assays, including hypersensitivity in response to intraplantar substance P, pain behaviours in response to intrathecal substance P and pain behaviour in the abdominal constriction test. In the discovery cohort of radiographic knee OA, an intronic single nucleotide polymorphism at rs900414 was significantly associated with symptomatic OA. Replication in three additional cohorts confirmed that the minor allele at rs900414 was consistently increased among asymptomatic compared to symptomatic radiographic knee OA cases in all four cohorts. A fixed-effects meta-analysis yielded an OR=1.35 (95% CI 1.17 to 1.56; p=4.3×10(-5) and no significant between-study heterogeneity). Studies in mice revealed that Pcsk6 knockout mice were significantly protected against pain in a battery of algesiometric assays. These results suggest that a variant in PCSK6 is strongly associated with protection against pain in knee OA, offering some insight as to why, in the presence of the same structural damage, some individuals develop chronic pain and others are protected. Studies in Pcsk6 null mutant mice further implicate PACE4 in pain.


Comparison of two prognostic scores (BSI and FACED) in a Spanish cohort of adult patients with bronchiectasis and improvement of the FACED predictive capacity for exacerbations.
Bronchiectasis (BE) is a chronic and heterogeneous respiratory disease that requires a multidimensional scoring system to properly assess severity. The aim of this study was to compare the severity stratification by 2 validated scores (BSI and FACED) in a BE cohort and to determine their predictive capacity for exacerbations and hospitalizations. Moreover, we proposed a modified version of FACED which was created to better predict the risk of exacerbations in clinical practice. We performed a prospective cohort study including BE patients >18 years old with a follow-up period of 1-year. One-hundred eighty-two patients (40% males; mean age 68) were studied. Patients were stratified according to the number of exacerbations during the follow-up, and according to BSI and FACED scores. BSI classified most of our patients as severe 99 (54.4%) or moderate 47 (25.8%), while FACED mainly classified as mild 108 (59.3%) or moderate 61 (33.5%). BSI and FACED showed an area under ROC curve (AUC) for exacerbations of 0.808 and 0.734; and for hospitalizations (due to BE exacerbations) of 0.893 and 0.809, respectively. Subsequently, we modified FACED by adding previous exacerbations (Exa-FACED) and this new score classified patients as mild 48.4%, moderate 34.6% and severe 17.0%, with an improved AUC for exacerbations (0.760) and hospitalizations (0.820). Despite previous validations of BSI and FACED, they classified our patients very differently. As expected, FACED showed poor prognostic capacity for exacerbations. We support the Exa-FACED score to predict the risk future exacerbations for been easy to use in clinical practice.


[Social competence and ability to make decisions].
Social competence and judgement can be separated in a somatic and in a mental competence. In mental competence, the capacity to consent has to be distinguished from the capacity to judge legal or more complex matters. Autonomy of patients requests that the capacity of consent is given, even in the presence of considerable cognitive impairment. On the other hand, competence in legal matters may be not any longer given in the presence of light to moderate cognitive impairment. In the assessment of competence, it is important that the person has the ability to make a choice, the ability to give directives, and the ability to do for oneself. Cognitively impaired patients therefore require assistance and protection in legal matters already at an early stage of the disease.


[Effect of collagenase on neuromuscular transmission in the frog].
Modification of synaptic function was estimated through the action of 0.1% solution of collagenase on neuromuscular junction in frog. Electrophysiological experiments revealed a decrease of the e.p.p. quantum content on single nerve stimulation with collagenase. The latter or the product of its splitting modify the action zones of mediator secretion. The demyelination action of collagenase on nerve is discussed.


A Comparison of US and Canadian Osteoporosis Screening and Treatment Strategies in Postmenopausal Women.
The optimal approach to osteoporosis screening and treatment in postmenopausal women is unclear. We compared (i) the United States Preventive Services Task Force (USPSTF) and Osteoporosis Canada osteoporosis screening strategies; and (ii) the National Osteoporosis Foundation (NOF) and Canadian treatment strategies. We used data from the prospective Women's Health Initiative Observational Study and Clinical Trials of women aged 50 to 79 years at baseline (n = 117,707 followed for self-reported fractures; n = 8134 in bone mineral density [BMD] subset). We determined the yield of the screening and treatment strategies in identifying women who experienced major osteoporotic fractures (MOFs) during a 10-year follow-up. Among women aged 50 to 64 years, 23.1% of women were identified for BMD testing under the USPSTF strategy and 52.3% under the Canadian strategy. For women ≥65 years, 100% were identified for testing under the USPSTF and Canadian strategies, 35% to 74% were identified for treatment under NOF, and 16% to 37% were identified for treatment under CAROC (range among 5-year age subgroups). Among women who experienced MOF during follow-up, the USPSTF strategy identified 6.7% of women 50 to 54 years-old and 49.5% of women 60 to 64 years-old for BMD testing (versus 54.4% and 60.6% for the Canadian strategy, respectively). However, the specificity of the USPSTF strategy was higher than that of the Canadian strategy among women 50 to 64 years-old. Among women who experienced MOF during follow-up, sensitivity for identifying women as treatment candidates was lowest for both strategies in women aged 50 to 64 (NOF 10% to 38%; CAROC 1% to 15%) and maximal in 75-year-old to 79-year-old women (NOF 82.8%; 51.6% CAROC); specificity declined with advancing age and was lower with the NOF compared to the CAROC strategy. Among women aged 50 to 64 years, the screening and treatment strategies examined had low sensitivity for identifying those who subsequently experience MOF; sensitivity was higher among women ≥65 years than among younger women. New screening and treatment algorithms are needed. © 2018 American Society for Bone and Mineral Research.


Suicidal ideation among attendees of a West African HIV clinic.
The paucity of information on suicide and its related issues among people living with HIV/AIDS (PLWHA) can impair evidence guided intervention. This study was set to determine the prevalence of suicidal ideation and the associated risk factors among PLWHA. A total of 295 participants made up of HIV positive individuals were subjected to a sociodemographic/clinical profile questionnaire. This was followed by the 28-item General Health Questionnaire (GHQ-28), suicidal intention item from the Beck Depression Inventory (BDI) and World Health Organisation Quality of Life (WHOQOL)--Bref scale to assess emotional distress, suicidal ideation, and quality of life respectively. The prevalence of suicidal ideation among PLWHA was 13.6%; and being unmarried, poor medication adherence, and poorer quality of life were significantly associated (p < 0.05) with suicidal ideation; while unemployment (odds ratio [OR] = 0.200; 95% Confidence Interval [CI] = 0.084-0.476; P < 0.001), emotional distress (OR = 5.734; 95% CI = 2.119-15.519; P--0.001), religion (OR = 4.829; 95% CI = 1.639-14.231; P--0.004), HIV status non-disclosure (OR = 2.630; 95% CI = 1.015-6.809; P--0.046) and previous suicidal attempt (OR = 0.172, 95% CI = 0.042-0.705; P--0.014) were not only associated but predictive of suicidal ideation in PLWHA. These findings indicate a significant burden of suicidal ideation, and psychosocial with clinical factors constitute identifiable risk factors among PLWHA. The development of evidence guided preventive and treatment measures against suicide among PLWHA are implied.


Myosin: immunofluorescent localization in neuronal and glial cultures.
The distribution of intracellular myosin was studied by the double antibody immunofluorescence method in primary organotypic neuronal cultures and two established neuronal and glial cell lines. An array of parallel filaments aligned with the major cellular axis and a three-dimensional subsurface network were shown to react with two different myosin antibodies. The presence of myosin-rich filaments in regions known to contain actin filaments suggests that these proteins interact to generate the motive force in nonmuscle contractile systems.


A Th2-score in the tumor microenvironment as a predictive biomarker of response to Bacillus Calmette Guérin in patients with non-muscle invasive bladder carcinoma: A retrospective study.
Intravesical Bacillus Calmette Guerin (BCG) is the gold standard therapy for intermediate/high-risk non-muscle invasive bladder cancer (NMIBC). However, the response rate is ~60%, and 50% of non-responders will progress to muscle-invasive disease. BCG induces massive local infiltration of inflammatory cells (Th1) and ultimately cytotoxic tumor elimination. We searched for predictive biomarker of BCG response by analyzing tumor-infiltrating lymphocyte (TIL) polarization in the tumor microenvironment (TME) in pre-treatment biopsies. Pre-treatment biopsies from patients with NMIBC who received adequate intravesical instillation of BCG (n = 32) were evaluated retrospectively by immunohistochemistry. TME polarization was assessed by quantifying the T-Bet+ (Th1) and GATA-3+ (Th2) lymphocyte ratio (G/T), and the density and degranulation of EPX+ eosinophils. In addition, PD-1/PD-L1 staining was quantified. The results correlated with BCG response. In most non-responders, Th1/Th2 markers were compared in pre-and post-BCG biopsies. ORR was 65.6% in the study population. BCG responders had a higher G/T ratio and a greater number of degranulated EPX+ cells. Variables combined into a Th2-score showed a significant association with higher scores in responders (p = 0.027). A Th2-score cut-off value >48.1 allowed discrimination of responders with 91% sensitivity but lower specificity. Relapse-free survival was significantly associated with the Th2-score (p = 0.007). In post-BCG biopsies from recurring patients, TILs increased Th2-polarization, probably reflecting BCG failure to induce a pro-inflammatory status and, thus, a lack of response. PD-L1/PD-1 expression was not associated with the response to BCG. Our results support the hypothesis that a pre-existing Th2-polarized TME predicts a better response to BCG, assuming a reversion to Th1 polarization and antitumor activity.


Chryscandin, a novel peptidyl nucleoside antibiotic. II. Structure determination and synthesis.
The structure of chryscandin, a novel antifungal antibiotic, produced by Chrysosporium pannorum No. 4629 was deduced to be 1 from spectroscopic and chemical evidences. In order to confirm the structure and to determine the absolute configuration, the total synthesis of chryscandin was performed. D-Xylose was transformed into 10 in ten steps. After beta-glycosidation of silylated benzoyl adenine with 10, the resulting 11 was converted into 4, which was identical with the product obtained from chryscandin by alkaline hydrolysis. From the key intermediate 13, chryscandin was synthesized via peptide formation followed by removal of the protecting groups. Chryscandin (1) is the first naturally occurring nucleoside antibiotic possessing a 3-aminoribofuranuronic acid in the molecule.


Pain management for labor and delivery in the 90s.
Although variable, labor pain is among the most severe of pain syndromes, and has been described as severe to excruciating in 50 to 70 percent of primiparas. "Twilight sleep" or amnesia was commonly used in the first half of this century via potent intramuscular, intravenous and inhalational agents. Subsequently, epidural anesthesia, first caudal then lumbar was used which offered superior pain relief without clouding the sensorium. However, epidurals with local anesthetics also contributed to dense sensory and motor blocks which are not necessary for labor. Presently, both spinal and epidural opioids are used along with decreasing doses of local anesthetics, rendering the laboring patient a relatively pain-free labor but allowing her more mobility and control of her environment.


Construction, Validation and Sensitivity Analyses of a Job Exposure Matrix for Early Plutonium Workers at the Sellafield Nuclear Site, United Kingdom.
Plutonium is a radiologically significant alpha-particle emitter. The potential for adverse health effects from internal exposures due to plutonium intakes has been recognized since the 1940s. The workforce of the Sellafield nuclear facility (Cumbria, UK), includes one of the world's most important groups of plutonium-exposed workers for studying the potential health risks of this internal exposure. However, for several hundred workers employed at the start of plutonium work at the facility (1952-1963), historical monitoring records based on measurements of urinary excretion of plutonium are not sufficiently reliable to provide the accurate and unbiased exposure assessments needed for epidemiological studies. Consequently, these early workers have had to be excluded from such studies, significantly reducing their power. We constructed a population-specific quantitative job exposure matrix (JEM) to estimate the average intakes of "typical plutonium workers" in this period, from 1952-1963, and assessed its validity and sensitivity to exposure assessment decisions. We conducted internal cross-validation using an a priori 10% extracted sample to evaluate reliability of estimates, explored JEM sensitivity to assumptions in the exposure assessment, and assessed the impact of uncertainty in urinalysis measurements on the precision of annual intake estimates using Markov Chain Monte Carlo (MCMC) methodology. Pairwise correlations ( RP) of estimated (JEM) and measured (10% sample) annual intakes were moderate to high ( RP > 0.4) for 10 out of 13 JEM groups, while absolute differences were <20% for 11 out of 13 JEM groups. There was little evidence of a temporal trend in correlations ( P = 0.13) or absolute differences ( P = 0.34). The median JEM-derived cumulative intake of 95.2 (IQR, 55.0-130.0) Bq was comparable to those based on alternative assumptions in the exposure assessment (median range, 95.2-100.0 Bq; 75th percentiles, 130.0-146.0 Bq). Measurement error simulation resulted in a 40-60% reduced median cumulative intake but higher maximum cumulative intakes. The JEM finds a balance between reliability and precision that makes it useful for epidemiological purposes and is relatively insensitive to specific choices in the exposure assessment. This JEM will allow the inclusion of workers with longest follow-up and who could not be included up until now in epidemiological studies without introducing significant bias.


SMAD proteins directly suppress PAX2 transcription downstream of transforming growth factor-beta 1 (TGF-β1) signalling in renal cell carcinoma.
Canonical TGF-β1 signalling promotes tumor progression by facilitating invasion and metastasis, whereby release of TGF-β1, by (for example) infiltrating immune cells, induces epithelial to mesenchymal transition (EMT). PAX2, a member of the Paired box family of transcriptional regulators, is normally expressed during embryonic development, including in the kidney, where it promotes mesenchymal to epithelial transition (MET). PAX2 expression is silenced in many normal adult tissues. However, in contrast, PAX2 is expressed in several cancer types, including kidney, prostate, breast, and ovarian cancer. While multiple studies have implicated TGF-β superfamily members in modulating expression of Pax genes during embryonic development, few have investigated direct regulation of Pax gene expression by TGF-β1. Here we have investigated direct regulation of PAX2 expression by TGF-β1 in clear cell renal cell carcinoma (CC-RCC) cell lines. Treatment of PAX2-expressing 786-O and A498 CC-RCC cell lines with TGF-β1 resulted in inhibition of endogenous PAX2 mRNA and protein expression, as well as expression from transiently transfected PAX2 promoter constructs; this inhibition was abolished in the presence of expression of the inhibitory SMAD, SMAD7. Using ChIP-PCR we showed TGF-β1 treatment induced SMAD3 protein phosphorylation in 786-O cells, and direct SMAD3 binding to the human PAX2 promoter, which was inhibited by SMAD7 over-expression. Overall, these data suggest that canonical TGF-β signalling suppresses PAX2 transcription in CC-RCC cells due to the direct binding of SMAD proteins to the PAX2 promoter. These studies improve our understanding of tumor progression and epithelial to mesenchyme transition (EMT) in CC-RCC and in other PAX2-expressing cancer types.


Role of endoscopic ultrasonography in patients suspected of pancreatic cancer with negative helical MDCT scan.
In some patients suspected of pancreatic cancer, no mass can be detected by multidetector computed tomography (MDCT) scan as the cause of biliary obstruction. All patients suspected of pancreatic cancer between January 2007 and 2009 with a negative MDCT were identified from a database. MDCT was performed for suspected pancreatic cancer in 290 patients, and in 258 a pancreatic mass was found. MDCT failed to establish a diagnosis in 32 patients (11%). In 23 patients (74%) with complete endoscopic ultrasonography (EUS), the cause of the obstruction was correctly diagnosed. A mass in the pancreatic head was found in 15 patients; 13 patients had a malignant tumor, and 2 patients a benign cause of obstruction. Further, EUS diagnosed 3 patients with a superficial adenoma of the papilla and 8 patients with a benign cause of the obstruction. In 5 patients EUS could not detect the cause of obstruction but finally a pancreatic malignancy was diagnosed. The positive predictive value of EUS was 86% and the negative predictive value 63%. Accuracy of MDCT and EUS decreased in the presence of pancreatitis or a biliary endoprosthesis. In patients suspected of pancreatic cancer in whom MDCT fails to demonstrate the cause of obstructive jaundice, EUS identifies 74% of the underlying diseases correctly.


[Lipolytic and catalase activity of bacteria-destructors of protective coatings].
Lipolytic and catalase activity of bacteria-destructors of protective coatings at different growth models: biofilm and plankton have been studied. It was shown that the synthesis of exoenzymes in the biofilm is different. Activity of extracellular lipase and catalase in the biofilm is 1.1-1.5 and 1.2-2.1 times higher, respectively, than in plankton. The experiment duration being increased to 14 days, the lipolytic activity considerably decreases. Activity of studied enzymes, produced by bacteria-destructors, can be used for estimating bioresistance of isolating coatings as the index of their biodegradation in technogenic media.


Splenic irradiation in the treatment of patients with chronic myelogenous leukemia or myelofibrosis with myeloid metaplasia. Results of daily and intermittent fractionation with and without concomitant hydroxyurea.
Seventeen patients with either chronic myelogenous leukemia (CML) or myelofibrosis with myeloid metaplasia (MMM) received 24 courses of splenic irradiation at this institution from 1973 to 1982. Eleven of the 17 patients had received prior chemotherapy. Patients were treated with 60Co gamma rays or 6 MV photons. The fraction size ranged from 15 to 100 rad and the total dose per treatment course from 15 to 650 rad, with the exception of one patient who received 1650 rad. Fourteen of 19 courses (71%) given for splenic pain yielded significant subjective relief while 17 of 26 courses given for splenomegaly obtained at least 50% regression of splenic size. Blood counts were carefully monitored before each treatment to limit hematologic toxicity. From this experience, the authors conclude that splenic irradiation effectively palliates splenic pain and reverses splenomegaly in the majority of patients with CML and MMM. Intermittent fractionation (twice or thrice weekly) is more convenient for the patient, appears to be as effective as daily treatment, and may be associated with less hematologic toxicity. Preliminary results of concurrent treatment with splenic irradiation and oral hydroxyurea show promise and warrant further study.


Exome Sequencing in a Swiss Childhood Glaucoma Cohort Reveals CYP1B1 and FOXC1 Variants as Most Frequent Causes.
The aim of this study was to investigate the molecular basis of childhood glaucoma in Switzerland to recommend future targeted genetic analysis in the Swiss population. Whole-exome sequencing and copy number variation (CNV) analysis was performed in a Swiss cohort of 18 patients from 14 unrelated families. Identified variants were validated by Sanger sequencing and multiplex ligation-dependent probe amplification. Breakpoints of structural variants were determined by a microarray. A minigene assay was conducted for functional analysis of a splice site variant. A diagnosis of primary congenital glaucoma was made in 14 patients, of which six (43%) harbored pathogenic variants in CYP1B1, one (7%) a frameshift variant in FOXC1, and seven (50%) remained without a genetic diagnosis. Three patients were diagnosed with glaucoma associated with nonacquired ocular anomalies, of which two patients with mild ocular features of Axenfeld-Rieger syndrome harbored a FOXC1 duplication plus an additional FOXC1 missense variant, and one patient with a Barkan membrane remained without genetic diagnosis. A diagnosis of juvenile open-angle glaucoma was made in one patient, and genetic analysis revealed a FOXC1 duplication. Sequencing of CYP1B1 and FOXC1, as well as analysis of CNVs in FOXC1, should be performed before extended gene panel sequencing. The identification of the molecular cause of childhood glaucoma is a prerequisite for genetic counseling and personalized care for patients and families.


A Drosophila glial cell atlas reveals a mismatch between transcriptional and morphological diversity.
Morphology is a defining feature of neuronal identity. Like neurons, glia display diverse morphologies, both across and within glial classes, but are also known to be morphologically plastic. Here, we explored the relationship between glial morphology and transcriptional signature using the Drosophila central nervous system (CNS), where glia are categorised into 5 main classes (outer and inner surface glia, cortex glia, ensheathing glia, and astrocytes), which show within-class morphological diversity. We analysed and validated single-cell RNA sequencing data of Drosophila glia in 2 well-characterised tissues from distinct developmental stages, containing distinct circuit types: the embryonic ventral nerve cord (VNC) (motor) and the adult optic lobes (sensory). Our analysis identified a new morphologically and transcriptionally distinct surface glial population in the VNC. However, many glial morphological categories could not be distinguished transcriptionally, and indeed, embryonic and adult astrocytes were transcriptionally analogous despite differences in developmental stage and circuit type. While we did detect extensive within-class transcriptomic diversity for optic lobe glia, this could be explained entirely by glial residence in the most superficial neuropil (lamina) and an associated enrichment for immune-related gene expression. In summary, we generated a single-cell transcriptomic atlas of glia in Drosophila, and our extensive in vivo validation revealed that glia exhibit more diversity at the morphological level than was detectable at the transcriptional level. This atlas will serve as a resource for the community to probe glial diversity and function.


A Lotus Seedpods-Inspired Interfacial Solar Steam Generator with Outstanding Salt Tolerance and Mechanical Properties for Efficient and Stable Seawater Desalination.
Interfacial solar steam generators (ISSGs) can capture solar energy and concentrate the heat at the gas-liquid interface, resulting in efficient water evaporation. However, traditional ISSGs have limitations in long-term seawater desalination processes, such as limited light absorption area, slow water transport speed, severe surface salt accumulation, and weak mechanical performance. Inspired by lotus seedpods, a novel ISSG (rGO-SA-PSF) is developed by treating a 3D warp-knitted spacer fabric with plasma (PSF) and combining it with sodium alginate (SA) and reduces graphene oxide (rGO). The rGO-SA-PSF utilizes a core-suction effect to achieve rapid water pumping and employs aerogel to encapsulate the plasma-treated spacer yarns to create the lotus seedpod-inspired hydrophilic stems, innovatively constructing multiple directional water transport channels. Simultaneously, the large holes of rGO-SA-PSF on the upper layer form lotus seedpod-inspired head concave holes, enabling efficient light capture. Under 1 kW m-2 illumination, rGO-SA-PSF exhibits a rapid evaporation rate of 1.85 kg m-2 h-1 , with an efficiency of 96.4%. Additionally, it shows superior salt tolerance (with no salt accumulation during continuous evaporation for 10 h in 10% brine) and self-desalination performance during long-term seawater desalination processes. This biomimetic ISSG offers a promising solution for efficient and stable seawater desalination and wastewater purification.


Toward Comprehensive Patient-Centric Care by Integrating Digital Health Technology With Direct Clinical Contact in Australia.
There is an escalating crisis in health care, locally and internationally. The current health care model is unable to meet the increasing health care demands. The aim of this study was to reconceptualize the provision of health care to produce better outcomes at no greater cost, by placing individuals in the position of authority to direct their own care, in a personalized, integrated health care system. In this study, we used the Australian health care system as a model. We reviewed the current landscape of digital health in Australia and discussed how electronic medical records (EMRs) can be further developed into a personalized, integrated health care system. Some components of an EMR and digital health system are already being used in Australia, but the systems are not linked. A personalized, integrated health care model that is responsive to consumer needs requires not just a passive repository of medical information; it would require a team approach, including the government, health care funders, industries, consumers and advocacy groups, health care professionals, community groups, and universities. Implementation of a personalized, integrated health care system can result in reduced pressure on the current health care system, and it can result in the delivery of best-practice health care, regardless of location. Importantly, a personalized, integrated health care system could serve as an education platform, "upskilling" not only clinicians but also, more importantly, patients and carers by providing them with accurate information about their condition, treatment options, medications, and management strategies. By proposing personalized, integrated health care, we offer an intelligent model of health care that is ubiquitous, efficient, and continuously improving.


An integrated research framework combining genomics, systems biology, physiology, modelling and breeding for legume improvement in response to elevated CO2 under climate change scenario.
How unprecedented changes in climatic conditions will impact yield and productivity of some crops and their response to existing stresses, abiotic and biotic interactions is a key global concern. Climate change can also alter natural species' abundance and distribution or favor invasive species, which in turn can modify ecosystem dynamics and the provisioning of ecosystem services. Basic anatomical differences in C3 and C4 plants lead to their varied responses to climate variations. In plants having a C3 pathway of photosynthesis, increased atmospheric carbon dioxide (CO2) positively regulates photosynthetic carbon (C) assimilation and depresses photorespiration. Legumes being C3 plants, they may be in a favorable position to increase biomass and yield through various strategies. This paper comprehensively presents recent progress made in the physiological and molecular attributes in plants with special emphasis on legumes under elevated CO2 conditions in a climate change scenario. A strategic research framework for future action integrating genomics, systems biology, physiology and crop modelling approaches to cope with changing climate is also discussed. Advances in sequencing and phenotyping methodologies make it possible to use vast genetic and genomic resources by deploying high resolution phenotyping coupled with high throughput multi-omics approaches for trait improvement. Integrated crop modelling studies focusing on farming systems design and management, prediction of climate impacts and disease forecasting may also help in planning adaptation. Hence, an integrated research framework combining genomics, plant molecular physiology, crop breeding, systems biology and integrated crop-soil-climate modelling will be very effective to cope with climate change.


Salvianolic acids attenuate rat hippocampal injury after acute CO poisoning by improving blood flow properties.
Carbon monoxide (CO) poisoning causes the major injury and death due to poisoning worldwide. The most severe damage via CO poisoning is brain injury and mortality. Delayed encephalopathy after acute CO poisoning (DEACMP) occurs in forty percent of the survivors of acute CO exposure. But the pathological cause for DEACMP is not well understood. And the corresponding therapy is not well developed. In order to investigate the effects of salvianolic acid (SA) on brain injury caused by CO exposure from the view point of hemorheology, we employed a rat model and studied the dynamic of blood changes in the hemorheological and coagulative properties over acute CO exposure. Compared with the groups of CO and 20% mannitol + CO treatments, the severe hippocampal injury caused by acute CO exposure was prevented by SA treatment. These protective effects were associated with the retaining level of hematocrit (Hct), plasma viscosity, fibrinogen, whole blood viscosities and malondialdehyde (MDA) levels in red blood cells (RBCs). These results indicated that SA treatment could significantly improve the deformation of erythrocytes and prevent the damage caused by CO poisoning. Meanwhile, hemorheological indexes are good indicators for monitoring the pathological dynamic after acute CO poisoning.


Multidimensional evaluation of tracheobronchial disease in adults.
The large airways can be affected by a wide spectrum of acquired benign and malignant diseases. These lesions may present as focal or diffuse processes and with narrowing or widening of the airway. Some of these may be asymptomatic for quite some time and may be incidentally detected on imaging, while others may be symptomatic, causing airway compromise. There may be a characteristic radiograph and computed tomography (CT) appearance, suggesting a narrow differential. When the imaging findings are not definitive, tissue may be obtained for pathological analysis. It behooves the radiologist to be familiar with the pathologic findings that correlate with the radiographic or CT appearance of the most frequently seen large airway lesions. In this way, we may improve our diagnostic accuracy. This paper will present the imaging findings of the most prevalent tracheobronchial lesions along with any associated pathology. Teaching Points • The large airways can be affected by many acquired benign and malignant diseases.• Large airway lesions may present as focal or diffuse processes, with narrowing or widening.• There may or may not be characteristic imaging appearance of large airway disease.• If imaging findings are not definitive, tissue may be obtained for pathological analysis.


Hairlessness in Mus musculus. A pathologic study.
The homozygous albino hairless mice at birth have hair bulbs and follicles the same as normal mice. Hairs develop and grow the same as in normal mice. However, about the sixth to eighth day hairs in the skin around the nose begin to disappear. This depletion of hairs continues over the head, neck, back, and abdomen of all mice in the same pattern. Essentially every hair over the entire body is absent macroscopically by the 30th to 40th day except for the vibrissae. A progressive atrophy of the hair bulbs and follicles is the basis for the absence of hairs after the first pelage. Only a rare follicle with a hair is present histologically in the adult mice. There are a few cysts in the dermis resulting from dilated residual hair follicles.


Direct and indirect cost of managing alzheimer's disease and related dementias in the United States.
Care of individuals with Alzheimer's Disease and Related Dementias (ADRD) poses special challenges. As the disease progresses, individuals with ADRD require increasing levels of medical care, caregiver support, and long-term care which can lead to substantial economic burden. Areas covered: In this expert review, we synthesized findings from studies of costs of ADRD in the United States that were published between January 2006 and February 2017, highlighted major sources of variation in costs, identified knowledge gaps and briefly outlined directions for future research and implications for policy and program planning. Expert commentary: A consistent finding of all studies comparing individuals with and without ADRD is that the average medical, non-medical, and indirect costs of individuals with ADRD are higher than those without ADRD, despite the differences in the methods of identifying ADRD, duration of the study, payer type and settings of study population. The economic burden of ADRD may be underestimated because many components such as direct non-medical costs for home safety modifications and adult day care services and indirect costs due to the adverse impact of ADRD on caregivers' health and productivity are not included in cost estimates.


Bioengineering of the Marine Diatom Phaeodactylum tricornutum with Cannabis Genes Enables the Production of the Cannabinoid Precursor, Olivetolic Acid.
The increasing demand for novel natural compounds has prompted the exploration of innovative approaches in bioengineering. This study investigates the bioengineering potential of the marine diatom Phaeodactylum tricornutum through the introduction of cannabis genes, specifically, tetraketide synthase (TKS), and olivetolic acid cyclase (OAC), for the production of the cannabinoid precursor, olivetolic acid (OA). P. tricornutum is a promising biotechnological platform due to its fast growth rate, amenability to genetic manipulation, and ability to produce valuable compounds. Through genetic engineering techniques, we successfully integrated the cannabis genes TKS and OAC into the diatom. P. tricornutum transconjugants expressing these genes showed the production of the recombinant TKS and OAC enzymes, detected via Western blot analysis, and the production of cannabinoids precursor (OA) detected using the HPLC/UV spectrum when compared to the wild-type strain. Quantitative analysis revealed significant olivetolic acid accumulation (0.6-2.6 mg/L), demonstrating the successful integration and functionality of the heterologous genes. Furthermore, the introduction of TKS and OAC genes led to the synthesis of novel molecules, potentially expanding the repertoire of bioactive compounds accessible through diatom-based biotechnology. This study demonstrates the successful bioengineering of P. tricornutum with cannabis genes, enabling the production of OA as a precursor for cannabinoid production and the synthesis of novel molecules with potential pharmaceutical applications.


AMH as the primary marker for fertility.
In women, the anti-Müllerian hormone (AMH) is secreted by the granulosa cells of growing follicles. Its measurement is strongly correlated with antral follicle count and represents a reliable marker of ovarian reserve. It also has the advantage of being highly reproducible since it has little variation within and between cycles. However, although it seems to be a good quantitative reflection of the ovarian reserve, it does not assess oocyte or embryo quality. This drawback precludes any good prediction of female fertility in the general population as well as in specific subgroups of patients. However, the AMH assay can become an indirect marker of the remaining female fertile years in some cases such as in those women who are at risk for premature ovarian failure or in those suffering from polycystic ovary syndrome. Its interest is no more to be proven in assisted reproductive technology where it is a valuable aid to the choice of the proposed techniques, ovarian stimulation protocols and gonadotropin dosage. AMH is finally very informative in monitoring cancer patients having received gonadotoxic drugs or having undergone mutilating ovarian surgeries. In conclusion, although it cannot be considered as a reliable predictor of pregnancy in women, AMH is now a useful tool in the management and treatment of female infertility.


Aspects of childhood cancer during the Byzantine period.
The main trends in the diagnosis and management of childhood cancer during the Byzantine period (330-1453 CE) are investigated. Therapeutic modalities reflected the influences from Ancient Greek and Greco-Roman medicine. Medical treatment included a great variety of regimens, and surgery was not unknown. The attitudes toward cancer suggest that people of that time did not believe in a superstitious origin of the disease. Even though most of these remedies and many procedures are nowadays out of use, the physicians of the Byzantine period preserved the scientific medical thought of antiquity, improved it, and set the basis of current achievements. Medical terms introduced during the Byzantine period are still used. The texts have been studied in their original languages, that is, Ancient and Byzantine Greek, and Latin.


Access to HIV care in jails: Perspectives from people living with HIV in North Carolina.
Most incarcerations for people living with HIV (PLWH) occur in jails, yet studies of HIV care during jail incarceration are limited. As part of a larger study to explore the ethical considerations in extending public health HIV surveillance to jail settings, we conducted semi-structured interviews with twenty-three PLWH with more than 300 distinct jail incarcerations post HIV diagnosis in 21 unique North Carolina jails. Interviews included questions about HIV disclosure in jail, the type of HIV care received in jail, and overall experiences with HIV care in jail. We report on participants' experiences and perspectives in four domains: access to HIV care in jail; impact of jail incarceration on continuity of HIV care; privacy and stigma; and satisfaction with HIV care in jail. Although most participants received HIV medications and saw providers while in jail, almost half reported that their greatest challenge in regard to HIV care was obtaining their HIV medications in the face of limited jail resources or policies that made access to medications difficult. Findings from this study suggest that jail leadership should review internal policies regarding HIV medications to ensure that PLWH can receive them quickly upon entry into jail. Findings also suggest that more external resources are needed, for example from state and local health departments, so that jails can provide timely HIV medications for PLWH incarcerated in their facilities.


Differential effect of cold acclimation on blood composition in rats and hamsters.
Male rats and hamsters were exposed to a progressively lower air temperature and shorter photoperiod to simulate the onset of winter. Normothermic hamsters had a higher haematological oxygen carrying capacity (OCC) and coagulability (shorter prothrombin time and activated partial thromboplastin time) than rats. Cold acclimation significantly increased the OCC of rats, which parallels an increased metabolic rate, while no differences were observed in hamsters. Red cell transit time through filters was faster in the acclimated rats but not in hamsters, reflecting the lower mean cell volume due to a decreased rate of clearance from the circulation. Platelet counts were significantly lower in both cold-acclimated rats and hamsters, and there was a significant leucopenia in rats, which would reduce the degree of microvascular blockade. Whole blood viscosity, plasma viscosity, and serum osmolarity showed little change in either species. However, whole blood viscosity was significantly lower in cold-acclimated hamsters than control hamsters at the lowest shear rate tested (0.95 s(-1)). Interestingly, plasma viscosity and serum osmolarity were significantly lower in hamsters exposed to low temperatures for a shorter period (4 weeks), and may reflect the development of a reduced coagulability. These data suggest that blood composition in hamsters contributes to an innate tolerance of low temperatures, maintaining tissue perfusion under hypothermic conditions and aiding arousal from hibernation.


Electrocardiographic changes in serious decompression sickness.
Electrocardiographic changes observed in 21 dogs suffering from spinal cord decompression sickness (DCS) are described. Changes seen included P wave peaking and P-R depression compatible with right heart strain; S-T segment and T wave changes suggestive of myocardial ischemia; and ventricular arrhythmias ranging from unifocal premature ventricular contractions to ventricular tachycardia. Compression therapy did not always restore the ECG changes promptly to normality. The changes are discussed in association with concurrent physiological events. These included pulmonary hypertension, systemic hypertension and hypotension, and cerebral DCS. Possible mechanisms ranging from local cardiac DCS or coronary gas embolism to autonomic nervous system disturbances arising from cerebral and spinal cord DCS are reviewed. It is concluded that ECG recordings should be made more often when treating clinical DCS.


A complex intervention to promote prevention of delirium in older adults by targeting caregiver's participation during and after hospital discharge - study protocol of the TRAnsport and DElirium in older people (TRADE) project.
Among potentially modifiable risk factors for delirium, transfers between wards, hospitals and other facilities have been mentioned with low evidence. TRADE (TRAnsport and DElirium in older people) was set up to investigate i) the impact of transfer and/or discharge on the onset of delirium in older adults and ii) feasibility and acceptance of a developed complex intervention targeting caregiver's participation during and after hospital discharge or transfer on cognition and the onset of delirium in older adults. The study is designed according to the guidelines of the UK Medical Research Council (MRC) for development and evaluation of complex interventions and comprises two steps: development and feasibility/piloting. The development phase includes i) a multicenter observational prospective cohort study to assess delirium incidence and cognitive decline associated with transfer and discharge, ii) a systematic review of the literature, iii) stakeholder focus group interviews and iv) an expert workshop followed by a Delphi survey. Based on this information, a complex intervention to better and systematically involve family caregivers in discharge and transport was developed. The intervention will be tested in a pilot study using a stepped wedge design with a detailed process and health economic evaluation. The study is conducted at four acute care hospitals in southwest Germany. Primary endpoints are the delirium incidence and cognitive function. Secondary endpoints include prevalence of caregiver companionship, functional decline, cost and cost effectiveness, quality of discharge management and quality of admission management in admitting hospitals or nursing homes. Data will be collected prior to discharge as well as after 3, 7 and 90 days. TRADE will help to evaluate transfer and discharge as a possible risk factor for delirium. In addition, TRADE evaluates the impact and modifiability of caregiver's participation during patient's transfer or discharge on delirium incidence and cognitive decline providing the foundation for a confirmatory implementation study. DRKS (Deutsches Register für klinische Studien) DRKS00017828 . Registered on 17th September 2019. Retrospectively registered.


Clinical features of bacterial meningitis in Italy: a multicenter prospective observational study.
We carried out a prospective observational study on clinical features of bacterial meningitis. Between October 2002 and June 2005, 322 adult bacterial meningitis cases in 49 infectious disease wards in Italy (MENTORE study group) were enrolled in the study. 133 cases were due to Streptococcus pneumoniae, 44 to Neisseria meningitidis and 145 to other microorganisms. A high SAPS score and coma on admission, as well as need for mechanical ventilation, were more frequent in the pneumococcal meningitis group. Neurological impairment was present in 151 out of 311 patients, and was more frequent in pneumococcal meningitis. A single antibiotic was employed in only 90 of 315 cases; a combination of ceftriaxone and ampicillin was the most frequently administered treatment. Ceftriaxone was also the single most used drug. Adjunctive treatment with steroids was administered in 210 out of 303 patients for a median duration of 7 days. Median duration of fever was 4 days, and median hospital stay was 16 days; hospitalization was significantly longer in the pneumococcal meningitis group. At discharge, neurological impairment was still present in 59 (21%) of 277 patients. Twenty (6.9%) out of 289 patients died during hospitalization. Distribution of adverse outcome (death and neurological impairment) in patients treated with or without steroids and within different time zones between onset of symptoms and commencement of antibiotics was studied; a trend toward a worse prognosis was seen in patients treated more than 24 hours after onset of the disease.In our study, infectious disease clinicians made extensive use of steroids as adjuvant therapy for bacterial meningitis, even in absence of detailed national and local guidelines. Mortality seemed to be lower in comparison with the literature.


Synthesis and molecular docking simulation of new benzimidazole-thiazole hybrids as cholinesterase inhibitors.
Dementia is a cognitive disturbance that is generally correlated with central nervous system diseases, especially Alzheimer's disease. The limited number of medications available is insufficient to improve the lifestyle of the patients suffering from this disease. Thus, new benzimidazole-thiazole hybrids (3-10) were designed and synthesized as acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory agents. The in vitro evaluation displayed that the derivatives 4b, 4d, 5b, 6a, 7a, and 8b demonstrated dual inhibitory efficiency against both AChE with IC50 ranging from 4.55 to 8.62 µM and BChE with IC50 ranging from 3.50 to 8.32 µM. By analyzing the Lineweaver-Burk plot, an uncompetitive form of inhibition was determined for the highly active compound 4d, revealing its inhibition type. The human telomerase reverse transcriptase-immortalized retinal pigment epithelial cell line was used to ensure the safety of the most potent cholinesterase inhibitors. Furthermore, compounds 4b, 4d, 5b, 6a, 7a, and 8b were evaluated for their neuroprotective and antioxidant properties, as well as their ability to suppress COX-2. The results demonstrated that compounds 4d, 5b, and 8b presented significant neuroprotection efficiency against H2 O2 -induced damage in SH-SY5Y cells with % cell viability of 67.42 ± 7.90%, 62.51 ± 6.71%, and 72.61 ± 8.10%, respectively, while the tested candidates did not reveal significant antioxidant activity. Otherwise, compounds 4b, 6a, 7a, and 8b displayed outstanding COX-2 inhibition effects with IC50 ranging from 0.050 to 0.080 μM relative to celecoxib (IC50 = 0.050 µM). In addition, molecular docking was carried out for the potent benzimidazole-thiazole hybrids with the active sites of both AChE (PDB ID: 4EY7) and BChE (PDB code: 1P0P). The tested candidates fit well in the active sites of both portions, with docking scores ranging from -8.65 to -6.64 kcal/mol (for AChE) and -8.71 to -7.73 kcal/mol (for BChE). In silico results show that the synthesized benzimidazole-thiazole hybrids have good physicochemical and pharmacokinetic properties with no Lipinski rule violations. The preceding results exhibited that compound 4d could be used as a new template for developing more significant cholinesterase inhibitors in the future.


Efficacy and safety of the haemostasis achieved by Vivostat system during laparoscopic partial nephrectomy.
Haemostasis remains the greatest challenge during laparoscopic partial nephrectomy. We describe the use of the Vivostat system helping effective haemostasis during laparoscopic partial nephrectomy (LPN). Twenty-eight patients underwent LPN. Autologous fibrin sealant was prepared with the Vivostat system and applied to the resection bed. This system is an automated medical device for the preparation of an autologous fibrin sealant from the patient's blood. Pre and postoperative clinical parameters and laboratory values were evaluated, for acute and delayed bleeding. Median patient's age was 58 years (range, 25-75). All patients underwent LPN for renal tumors (mean size 2.5 cm; range 0.9-4.5 cm). Six resection were performed without vessels clamping, and 22 were realized with selective arterial Bulldog clamping. Haemostasis was achieved by a cellulose bolster (80%), by stitches (67%) and by sealant application after declamping (100%) (mean amount applied: 5.1 ml). The mean warm ischemia time was 26 minutes (range, 16-45) for 22 interventions. Mean blood loss was 128 cc (range, 20-500). Pre-operative and post-operative creatinine values (mean, 0.91 vs. 1 ng/mi) did not differ significantly; whereas mean Hb levels slightly decreases after surgery (mean, 14.7 vs. 12.5 g/dl). Mean operative time was 131 minutes (range, 60-190). All but one had negative surgical margins. One intraoperative bleeding occurred needing blood transfusion (1 unit). Postoperatively, we observed only 1 perirenal hematoma treated conservatively requiring blood transfusion. In this study, an effective haemostasis was achieved and maintained after kidney reperfusion. These data support the previous finding with the same system and encourage its use in LPN.


Microscopic examination and broth culture of cerebrospinal fluid in diagnosis of meningitis.
We reviewed the results of microscopic Gram stain examination and routine culture for 2,635 cerebrospinal fluid (CSF) samples processed in an adult hospital microbiology laboratory during 55 months. There were 56 instances of bacterial or fungal meningitis (16 associated with central nervous system [CNS] shunt infection), four infections adjacent to the subarachnoid space, four cases of sepsis without meningitis, and an additional 220 CSF specimens with positive cultures in which the organism isolated was judged to be a contaminant. Because 121 of these contaminants were isolated in broth only, elimination of the broth culture would decrease unnecessary work. However, 25% of the meningitis associated with CNS shunts would have been missed by this practice. The most common cause of meningitis was Cryptococcus neoformans, followed by Streptococcus pneumoniae and Neisseria meningitidis. In 48 of 56 (88%) of cases, examination of the Gram-stained specimen revealed the causative organism. If patients who had received effective antimicrobial therapy prior to lumbar puncture are excluded, the CSF Gram stain is 92% sensitive. Microscopic examination incorrectly suggested the presence of organisms in only 3 of 2,635 (0.1%) CSF examinations. Thus, microscopic examination of Gram-stained, concentrated CSF is highly sensitive and specific in early diagnosis of bacterial or fungal meningitis.


Cognitive subtypes in recent onset psychosis: distinct neurobiological fingerprints?
In schizophrenia, neurocognitive subtypes can be distinguished based on cognitive performance and they are associated with neuroanatomical alterations. We investigated the existence of cognitive subtypes in shortly medicated recent onset psychosis patients, their underlying gray matter volume patterns and clinical characteristics. We used a K-means algorithm to cluster 108 psychosis patients from the multi-site EU PRONIA (Prognostic tools for early psychosis management) study based on cognitive performance and validated the solution independently (N = 53). Cognitive subgroups and healthy controls (HC; n = 195) were classified based on gray matter volume (GMV) using Support Vector Machine classification. A cognitively spared (N = 67) and impaired (N = 41) subgroup were revealed and partially independently validated (Nspared = 40, Nimpaired = 13). Impaired patients showed significantly increased negative symptomatology (pfdr = 0.003), reduced cognitive performance (pfdr < 0.001) and general functioning (pfdr < 0.035) in comparison to spared patients. Neurocognitive deficits of the impaired subgroup persist in both discovery and validation sample across several domains, including verbal memory and processing speed. A GMV pattern (balanced accuracy = 60.1%, p = 0.01) separating impaired patients from HC revealed increases and decreases across several fronto-temporal-parietal brain areas, including basal ganglia and cerebellum. Cognitive and functional disturbances alongside brain morphological changes in the impaired subgroup are consistent with a neurodevelopmental origin of psychosis. Our findings emphasize the relevance of tailored intervention early in the course of psychosis for patients suffering from the likely stronger neurodevelopmental character of the disease.


Good correlation between the Afinion AS100 analyser and the ABX Pentra 400 analyser for the measurement of glycosylated haemoglobin and lipid levels in older adults in Durban, South Africa.
The Afinion AS100 analyser is a small bench-top, multi-assay, point-of-care (POC) analyser that is able to measure glycated haemoglobin (HbA1c) and lipid levels. To assess performance of the Afinion analyser compared with a reference laboratory test for the measurement of HbA1c and lipid levels. The study involved men and women enrolled in a cross-sectional study, Sexual health, HIV infection and comorbidity with non-communicable diseases among Older Persons (SHIOP), which was conducted from February to May 2016. Whole blood was drawn aseptically by a trained study nurse into a serum separator gel tube and an ethylenediaminetetra-acetic acid (EDTA) tube. The EDTA whole blood was used to measure HbA1c levels, and serum to measure total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C) and triglyceride levels. Lin's correlation coefficient was used to assess the agreement between the Afinion and ABX Pentra 400 analysers for each marker. A total of 435 older individuals were included in the study. The proportion of HbA1c results that were correctly classified by the Afinion analyser was 92.2%. Bland-Altman analysis and linear regression analysis showed a very good agreement (correlation concordance 0.89) between the two analysers for the measurement of HbA1c. The two-way scatter plot for TC showed a substantial correlation (0.80). However, a total of 69 cholesterol results that were within the normal range on the Pentra were misclassified as abnormal on the Afinion. The readings obtained for HDL-C levels with the Afinion were shown to be slightly overestimated when compared with the Pentra. However, correlation for HDL-C on the two analysers was 0.93, indicating an almost perfect agreement. Seventy-four LDL-C results were erroneously classified as abnormal on the Afinion but were within the normal range on the Pentra, resulting in a substantial correlation of 0.75. An excellent agreement was observed between triglyceride measurements (0.99). This study supports the use of the Afinion AS100 analyser in POC testing for the measurement of HbA1c, triglycerides and HDL-C in a South African setting.


Association between exhaled inflammatory markers and asthma control in children.
The relationship between exhaled inflammatory markers and asthma control in children is unclear. To explore the association between inflammatory markers in exhaled breath (fractional nitric oxide (FeNO), volatile organic compounds (VOCs), cytokines/chemokines) and asthma control. To assess whether exhaled inflammatory markers are able to discriminate between children with persistently controlled/uncontrolled asthma. 96 asthmatic children were followed-up in a one-year observational study. Every 2 months, the following parameters were assessed: asthma control, FeNO, lung function (forced expiratory volume in 1 s (FEV1) and forced vital capacity (FVC), exhaled VOCs, and cytokines/chemokines in exhaled breath condensate (EBC). Random Forest was used to analyse the relationship between exhaled inflammatory markers and asthma control. For each model, patients were randomly selected for a training set and validation set. To assess the accuracy of the classification models, receiver operating characteristic-curves (ROC-curves) were generated. No significant association was found between the exhaled inflammatory markers (FeNO, markers in EBC, VOCs) and asthma control (area under the ROC-curve 49%). However, 15 exhaled VOCs could discriminate between subgroups of children with persistently controlled and uncontrolled asthma during all clinical visits (area under the ROC-curve 86%). Adding FeNO and markers in EBC to this model, did not lead to a more accurate classification (area under the ROC-curve 87%). There was no association between exhaled inflammatory markers and asthma control in children. However, children with persistently controlled or uncontrolled asthma during the 12 month study period could be discriminated by a set of VOCs.


Analysis of the transcription factors and their regulatory roles during a step-by-step differentiation of induced pluripotent stem cells into hepatocyte-like cells.
We investigated the human induced pluripotent stem cells (hiPSCs) during a sequential in vitro step-by-step differentiation into hepatocyte-like cells (HLCs) using nanoCAGE, a method for promoters, transcription factors, and transcriptome analysis. Specific gene clusters reflected the different steps of the hepatic differentiation. The proliferation step was characterized by a typical cell cycle and DNA replication. The hepatic endoderm and the HLC steps were marked by a common signature including cell interactions with extracellular matrix (ECM), lipoproteins and hepatic biomarkers (such as albumin and alpha-fetoprotein). The specific HLC profile was characterized by important transcription factors such as HIF1A, JUN, MAF, KLF6, BMP4 and with a larger expression of genes related to Wnt signaling, extracellular matrix, lipid metabolism, urea cycle, drugs, and solute transporters. HLC profile was also characterized by the activation of upstream regulators such as HNF1A, MEIS2, NFIX, WRNIP1, SP4, TAL1. Their regulatory networks highlighted HNF4a as a bridge and linked them to important processes such as EMT-MET transitions, ECM remodeling and liver development pathways (HNF3, PPARA signaling, iron metabolism) along the different steps of differentiation.


Hidden electrophoretic variation at the xanthine dehydrogenase locus in a natural population of Drosophila melanogaster.
Sixty-two isochromosomal lines of D. melanogaster were screened for cryptic electrophoretic variation at the xanthine dehydrogenase (XDH) locus. Sequential polyacrylamide vertical slab gel electrophoresis was performed using four electrophoretic criteria. A total of 15 classes of electromorphs were revealed. D. melanogaster appears to exhibit as much polymorphism at this locus as other extensively studied Drosophila species.--No evidence for loci on the X or second chromosomes which modified XDH mobility was found. Six of the electromorphs were mapped to the Xdh (ry) structural locus. Eight of the remaining nine classes exhibited mobility variation consistent with structural variation at the Xdh locus. The final class exhibited aberrant patterns and is under further study.


FKBP10/FKBP65 expression in high-grade ovarian serous carcinoma and its association with patient outcome.
The frequent loss of chromosome 17 in epithelial ovarian carcinomas (EOC), particularly high-grade serous carcinomas (HGSC), has been attributed to the disruption of TP53 (at 17p13.1) and other chromosome 17 genes suspected to play a role in tumour suppressor pathways. In a transcriptome analysis of HGSC, we showed underexpression of a number of chromosome 17 genes, which included FKBP10 (at 17q21.1) and collagen I α 1 (COL1A1; at 17q21.33). FKBP10 codes for the immunophilin FKBP65 and is suspected to act as a chaperone for COL1A1. We have investigated FKBP10 (gene) and FKBP65 (protein) expression in HGSC samples and EOC cell lines that differ in their tumourigenic potential. COL1A1 expression was also investigated given the purported function of FKBP65. RT-PCR analysis verified underexpression of FKBP10 and COL1A1 in HGSCs (n=14) and six tumourigenic EOC cell lines, relative to normal ovarian surface epithelial cells and a non-tumourigenic EOC cell line. Immunohistochemistry analyses of 196 HGSC samples using tissue microarrays revealed variable staining intensities in the epithelial tumour component where only 7.8% and 1.0% of samples stained intensely for FKBP65 and COL1A1, respectively. Variable staining intensities were also observed for the stromal component where 23.6% and 24.1% stained intensely for FKBP65 and COL1A1, respectively. There was no significant correlation of staining intensity of either protein with disease stage. Staining of FKBP65 was clearly visible in normal epithelial cells of the ovarian surface and fallopian tube. There was a significant correlation between absence of FKBP65 staining in the epithelial cell component of the tumour and prolonged overall survival (p<0.001). Our results suggest that underexpression of FKBP65 protein is characteristic of HGSCs and that this expression profile may be linked to molecular pathways associated with an unfavourable outcome in cancer patients.


Technical Feasibility of Tissue Microarray (TMA) Analysis of Tumor-Associated Immune Response in Prostate Cancer.
Introduction: The androgen receptor (AR) regulates immune-related epithelial-to-mesenchymal transition (EMT), and prostate cancer (PCa) metastasis. Primary tumor-infiltrating lymphocytes (TILs) [CD3+, CD4+, and CD8+ TILs] are potential prognostic indicators in PCa, and variations may contribute to racial disparities in tumor biology and PCa outcomes. Aim: To assess the technical feasibility of tumor microarray (TMA)-based methods to perform multi-marker TIL profiling in primary resected PCa. Methods: Paraffin-embedded tissue cores of histopathologically-confirmed primary PCa (n = 40; 1 TMA tissue specimen loss) were arrayed in triplicate on TMAs. Expression profiles of AR, CD3+, CD4+, and CD8+ TILs in normal prostate, and the center and periphery of both the tumor-dominant nodule and highest Gleason grade were detected by IHC and associated with clinical and pathological data using standard statistical methodology. An independent pathologist, blinded to the clinical data, scored all samples (percent and intensity of positive cells). Results: TMAs were constructed from 21 (53.8%) Black and 18 (46.2%) White males with completely-resected, primarily pT2 stage PCa [pT2a (n = 3; 7.7%); pT2b (n = 2; 5.1%); pT2c (n = 27; 69.2%); pT3a (n = 5; 12.8%); mean pre-op PSA = 8.17 ng/ml]. The CD3, CD4, CD8, and CD8/CD3 cellular protein expression differed from normal in the periphery of the dominant nodule, the center of the highest Gleason grade, and the periphery of the highest Gleason grade (P < 0.05). Correlations between TIL expression in the center and periphery of the dominant nodule, with corresponding center and periphery of the highest Gleason grade, respectively, were robust, and the magnitude of these correlations differed markedly by race (P < 0.05). Conclusions: Multi-marker (AR, CD3, CD4, CD8) profiling with IHC analysis of TMAs consisting of primary, non-metastatic resected prostate cancer is technically feasible in this pilot study. Future studies will evaluate primary tumor immunoscore using semi-quantitative, IHC-based methodology to assess differences in the spectrum, quantity, and/or localization of TILs, and to gain insights into racial disparities in PCa tumor biology and clinical outcomes.


Unfertilized Xenopus eggs die by Bad-dependent apoptosis under the control of Cdk1 and JNK.
Ovulated eggs possess maternal apoptotic execution machinery that is inhibited for a limited time. The fertilized eggs switch off this time bomb whereas aged unfertilized eggs and parthenogenetically activated eggs fail to stop the timer and die. To investigate the nature of the molecular clock that triggers the egg decision of committing suicide, we introduce here Xenopus eggs as an in vivo system for studying the death of unfertilized eggs. We report that after ovulation, a number of eggs remains in the female body where they die by apoptosis. Similarly, ovulated unfertilized eggs recovered in the external medium die within 72 h. We showed that the death process depends on both cytochrome c release and caspase activation. The apoptotic machinery is turned on during meiotic maturation, before fertilization. The death pathway is independent of ERK but relies on activating Bad phosphorylation through the control of both kinases Cdk1 and JNK. In conclusion, the default fate of an unfertilized Xenopus egg is to die by a mitochondrial dependent apoptosis activated during meiotic maturation.


Moonquakes: mechanisms and relation to tidal stresses.
Observed features of moonquakes are combined with theoretical calculations of the tidal stresses to interpret the moonquake mechanisms. Tidal stresses, together with a postulated ambient tectonic stress, are sufficient to explain the depth, periodicity, and polarity reversal of moonquakes. Both of these stresses are small (on the order of 1 bar) and consistent with the small magnitudes of moonquakes.


Infected Subgaleal Hematoma Following Blunt Head Trauma in a Child: Case Report and Review of the Literature.
Subgaleal hematoma (SGH), an uncommon but potentially dangerous complication, has been reported to occur with delivery in newborns, as well as in young patients following head trauma. Infection of a SGH is extremely rare, especially in cases where no disruption of the skin barrier occurs. We report a case of an infected SGH in an 8-month-old following closed skull fracture. The patient presented with scalp swelling 1 day after falling 3 feet. Initial evaluation found a nondisplaced skull fracture on computed tomography. She was discharged following an uneventful 23-hour observation. Three days later, she developed symptoms concerning for a viral upper respiratory tract infection and received symptomatic treatment. Nine days after injury, she returned with continued fevers, irritability, and significant increase in scalp swelling. Magnetic resonance imaging showed a subgaleal abscess with osteomyelitis. Needle aspiration revealed an infected hematoma with cultures positive for Streptococcus pneumoniae, treated with intravenous ampicillin. Purulent drainage from an enlarging necrotic needle aspiration site required subsequent surgical debridement of the subgaleal abscess with drain placement. She recovered well following surgery and intravenous antibiotics. Physicians should be aware that SGH carries a risk of serious morbidity and mortality. SGH can serve as a nidus for infection, typically from skin barrier breakdown or, as in this case, hematogenous spread. Early recognition, appropriate antibiotic therapy, and surgical debridement are critical in treating infected SGH.


Metabolism of amino acids differs in the brains of Djungarian hamster (P. sungorus) and Roborovskii hamster (P. roborovskii).
Djungarian hamster (P. sungorus) and Roborovskii hamster (P. roborovskii) belong to the same genus of phodopus. Roborovskii hamster shows high locomotor activity and low level of dopamine (DA) in the brain. Administration of L-tyrosine, a precursor of DA, decreases locomotor activity in Roborovskii hamsters. However, the amino acid metabolism in relation to the hyperactivity is not yet well known. In the present study, L- and D-amino acid concentrations in the brain, liver, and plasma in Djungarian and Roborovskii hamsters were investigated during day and night times to explain the possible difference in hyperactivity between them. Most of the examined amino acids were higher in the night time when hamsters are active compared to those in day time. L- and D-tyrosine concentrations were higher in the liver of Roborovskii hamsters than in Djungarian hamsters. Furthermore, brain concentration of D-tyrosine was higher in the Roborovskii than in Djungarian hamsters, but no significant difference was observed for L-tyrosine concentrations between the two species. These results suggest that the conversion of L-tyrosine to D-tyrosine in the brain of Roborovskii hamster may be higher than in Djungarian hamster, which may cause low DA concentration and hyperactivity in Roborovskii hamster. On the other hand, L- and D-serine, which are known as sedative factors, were lower in Roborovskii hamsters than Djungarian hamster. These results suggest that species-specific regulation in amino acid metabolism may contribute to hyperactivity in Roborovskii hamsters.


Surgical Treatment for Extratemporal Epilepsy.
Partial seizures of extratemporal origin may present unique challenges in the patient with medically refractory seizures. The efficacy of an extratemporal focal cortical resection may be less effective than an anterior temporal lobectomy for intractable epilepsy. The potential operative complications may be increased in individuals with extratemporal epilepsy because of functional cerebral cortex involvement and the need for a large cortical resection to significantly reduce seizure tendency. Partial seizures of extratemporal origin are predominantly associated with frontal lobe epilepsy. The most effective treatment for intractable partial epilepsy is a focal cortical resection with excision of the epileptogenic zone, that is, an area of ictal onset and initial seizure propagation. The preoperative evaluation and operative strategy in patients with partial epilepsy of extratemporal origin associated with pharmacoresistant seizures is determined by the anatomic localization of the epileptogenic zone and the presence of a substrate-directed disorder. The goals of surgical treatment in extratemporal epilepsy include rendering the patient seizure-free, avoiding operative morbidity, and allowing the individual to become a participating and productive member of society. Before surgical treatment, the individual with extratemporal epilepsy will require a comprehensive preoperative evaluation, including routine electroencephalogram (EEG), long-term EEG monitoring, neuropsychologic studies, and magnetic resonance imaging (MRI). Patients with a normal MRI study, conflicting preoperative evaluation, or involvement of suspected functional cerebral cortex would require chronic intracranial EEG monitoring. The rationale for intracranial EEG includes localization of the ictal onset zone or intraoperative functional mapping, or both. Two-fluorodeoxyglucose positron emission tomography studies are usually unremarkable in patients with extratemporal epilepsy and normal MRI scans. Subtraction ictal single photon emission computed tomography coregistered to MRI (SISCOM) study may be useful to demonstrate a localized cerebral perfusion alteration in patients with intractable partial epilepsy. The diagnostic yield of SISCOM has been confirmed in patients with extratemporal epilepsy undergoing surgical treatment. The results of the SISCOM study may tailor the placement of intracranial EEG electrodes and affect the operative strategy. Patients with extratemporal epilepsy overall are less favorable operative candidates than individuals with medial temporal lobe epilepsy. However, individuals with MRI-identified lesional pathology of SISCOM-identified perfusion alterations concordant with the epileptogenic zone may be considered for surgical treatment. Chronic intracranial EEG monitoring may be necessary to confirm the localization of the ictal onset zone before epilepsy surgery. Patients with normal neuroimaging studies and extratemporal epilepsy are unlikely to be rendered seizure-free with focal cortical resection and should be considered candidates for other alternative forms of treatment for intractable partial epilepsy. Patients with non-substrate-directed extratemporal epilepsy should undergo a preoperative evaluation and surgical treatment at a comprehensive epilepsy center with extensive experience in chronic intracranial EEG monitoring and contemporary neuroimaging procedures because of the inherently high acuity associated with the operative management clinical disorder.


Screening for depression in a diabetic outpatient population.
Depression occurs twice as often in patients with diabetes and is associated with reduced compliance with exercise, diet, and medications. It is also associated with hyperglycemia and increased diabetic complications. Despite evidence that successful treatment is associated with improved glycemic control, many cases of depression are left untreated. Objectives. (1) Evaluate a combination screening strategy in an outpatient population; and (2) explore the association between glycemic control and depressive symptomatology. Methods. Ninety-two patients completed the Patient Health Questionnaire (PHQ-2). Patients with a PHQ-2 score ≥ 1 completed the 16-item Quick Inventory of Depressive Symptomatology (QIDS-SR16). Using the QIDS-SR16, a score of ≤5 corresponded to normal mood, with scores above 5 corresponding to increasing severity of depressive symptoms. Glycemic control was assessed by glycosylated hemoglobin (HbA1c). Results. Using a PHQ-2 cut-off score of ≥3, 37% of the sample screened positive for major depressive disorder (MDD), with an additional 27% reporting sub-threshold symptoms. The depressed group reported significantly more difficulty with reduced interests, insomnia, concentration, self-criticism, energy/fatigue and depressed mood. In terms of glycemic control, there was a marginally significant effect for race and HbA1c. Conclusion. The combined PHQ-2 and QIDS-SR16 can facilitate prompt detection of MDD and provide a means of monitoring specific symptoms and progress once treatment commences.


Antigen stimulation of peripheral blood mononuclear cells from Mycobacterium bovis infected cattle yields evidence for a novel gene expression program.
Bovine tuberculosis (BTB) caused by Mycobacterium bovis continues to cause substantial losses to global agriculture and has significant repercussions for human health. The advent of high throughput genomics has facilitated large scale gene expression analyses that present a novel opportunity for revealing the molecular mechanisms underlying mycobacterial infection. Using this approach, we have previously shown that innate immune genes in peripheral blood mononuclear cells (PBMC) from BTB-infected animals are repressed in vivo in the absence of exogenous antigen stimulation. In the present study, we hypothesized that the PBMC from BTB-infected cattle would display a distinct gene expression program resulting from exposure to M. bovis. A functional genomics approach was used to examine the immune response of BTB-infected (n = 6) and healthy control (n = 6) cattle to stimulation with bovine tuberculin (purified protein derivative - PPD-b) in vitro. PBMC were harvested before, and at 3 h and 12 h post in vitro stimulation with bovine tuberculin. Gene expression changes were catalogued within each group using a reference hybridization design and a targeted immunospecific cDNA microarray platform (BOTL-5) with 4,800 spot features representing 1,391 genes. 250 gene spot features were significantly differentially expressed in BTB-infected animals at 3 h post-stimulation contrasting with only 88 gene spot features in the non-infected control animals (P < or = 0.05). At 12 h post-stimulation, 56 and 80 gene spot features were differentially expressed in both groups respectively. The results provided evidence of a proinflammatory gene expression profile in PBMC from BTB-infected animals in response to antigen stimulation. Furthermore, a common panel of eighteen genes, including transcription factors were significantly expressed in opposite directions in both groups. Real-time quantitative reverse transcription PCR (qRT-PCR) demonstrated that many innate immune genes, including components of the TLR pathway and cytokines were differentially expressed in BTB-infected (n = 8) versus control animals (n = 8) after stimulation with bovine tuberculin. The PBMC from BTB-infected animals exhibit different transcriptional profiles compared with PBMC from healthy control animals in response to M. bovis antigen stimulation, providing evidence of a novel gene expression program due to M. bovis exposure.


15,16-dihydrotanshinone I inhibits EOMA cells proliferation by interfering in posttranscriptional processing of hypoxia-inducible factor 1.
Infantile hemangioma (IH), which threatens the physical and mental health of patients, is the most common benign tumor in infants. Previously, we found that 15,16-dihydrotanshinone I (DHTS) was significantly more effective at inhibiting hemangioma proliferation in vitro and in vivo than the first-line treatment propranolol. To investigate the underlying mechanism of DHTS, we used EOMA cells as a model to study the effect of DHTS. We compared the transcriptomes of control and DHTS-treated EOMA cells. In total, 2462 differentially expressed genes were detected between the groups. Kyoto Encyclopedia of Genes and Genomes pathway analysis revealed downregulated activity of the hypoxia-inducible factor 1 alpha (HIF-1α) signaling pathway in EOMA cells following treatment with DHTS. Thus, we investigated HIF-1α expression at protein and mRNA levels. Our results revealed that DHTS downregulated HIF-1α expression by interfering in its posttranscriptional processing, and the RNA-binding protein HuR participated in this mechanism. Our findings provide a basis for clinical transformation of DHTS and insight into pathogenic mechanisms involved in IH.


Intracortical motor conduction is associated with hand dexterity in progressive multiple sclerosis.
Hand dexterity dysfunction is a key feature of disability in people with progressive multiple sclerosis (PMS). It underlies corticospinal tract (CST) and cerebellar integrity, as well as disruption of cortical networks, which are hardly assessed by standard techniques. Transcranial magnetic stimulation is a promising tool for evaluating the integrity of intracortical motor pathways. To investigate neurophysiological correlates of motor hand impairment in PMS. Antero-posterior (AP) stimulation of the primary motor cortex activates the CST indirectly through polysynaptic pathways, while a direct CST activation occurs with latero-medial (LM) directed current. Thirty PMS and 15 healthy controls underwent dominant hand motor evoked potentials (MEP) using AP and LM-directed stimulation, and a clinical assessment of dexterity (nine-hole peg test) and strength (MRC scale, grip and pinch). PMS with AP-LM latency difference 2.5 standard deviation above the mean of controls (33%) showed worse dexterity but no difference in upper limb strength. Accordingly, AP-LM latency shortening predicted dexterity (R2 = 0.538, p < 0.001), but not strength impairment. On the contrary, absolute MEP latencies only correlated with strength (grip: R2 = 0.381, p = 0.014; MRC: R2 = 0.184, p = 0.041). AP-LM latency shortening may be used to assess the integrity polysynaptic intracortical networks implicated in dexterity impairment.


Isometric Exercise Training: A Review of Hypothesized Mechanisms and Protocol Application in Persons with Hypertension.
According to the American Heart Association 116.4 million, or 46% of US adults are estimated to have hypertension. Although, traditional moderate intensity aerobic exercise training is associated with reducing blood pressure by 5-8 mmHg, barriers to this modality of exercise training exist. Thus, the purpose of this review is to evaluate the mechanisms and incorporation of isometric exercise training (IET) as an adjunctive mode of exercise in a population with HTN. Based upon the articles reviewed from the years 2000-2020 which incorporated IET and provided clear protocols lasting 4 or more weeks, meaningful reductions in blood pressure occurred following IET (SBP, -9.7 ± 3.3 mmHg; DBP, -4.8 ± 2.6 mmHg) which support the need to increase adoption of this exercise form into practice to help treat hypertension. Specifically, an IET program of 12-20 minutes per day, 3 times per week, could improve blood pressure reduction in those with hypertension. IET has the potential to produce significant and clinically meaningful blood pressure reductions and could serve as an adjunctive exercise modality alongside the established exercise prescription for those with hypertension.


Is Lyapunov exponent a reliable metric to detect dynamic stability in Parkinson's disease?
Idiopathic Parkinson's disease (PD) is the second most common neurodegenerative disorder worldwide. It affects the nervous system, causing motor and non-motor symptomatology. However, its clinical diagnosis remains dependent on the expertise of clinicians, as perceptual clinical scales are often used. Gait stability is one of the most relevant motor signs in PD. Nonetheless, it is usually not reported or quantified, possibly due to its unclear meaning and the high variability of metrics used in the literature. This work aims to identify a reliable and objective indicator that clinicians can use to assess patients in realistic contexts. We focused on the Largest Lyapunov Exponent (LLE), being the most common metric used in previous research works to quantify gait stability. The short and long-term LLEs were calculated in a group of 34 healthy and 42 participants diagnosed with PD. The long-term LLE extracted from the chest, right arm and right foot sensors showed statistical differences between subjects with PD and healthy control (HC) subjects, showing that the HC subjects are more stable than PD patients, whereas the short-term LLE showed the opposite results. Further investigation is required to clarify the reliability of this metric to detect and rate gait stability in people affected with PD.Clinical Relevance- This study is the first step towards the identification of an objective methodology to assess gait stability in clinical settings. Achieving this goal will contribute to improve the understanding and support the diagnosis of gait disorders that cause gait stability problems.


Movement of criticall ill patients within hospital.
Critically ill patients were observed during routine movement inside the hospital to and from the intensive therapy unit. One patient a month suffered major cardiorespiratory collapse or death as a direct result of movement. Renewed bleeding of a pelvic fracture, cardiac arrhythmia, cardiac embarrassment due to a haemothorax, and cardiovascular decompensation were seen. It was difficult to continue treatment during movement, especially maintaining an airway or providing adequate intermittent positive pressure ventilation. Seventy postoperative patients suffered few ill effects on being moved. Greater awareness of the dangers of moving critically ill patients within hospital is needed. Thorough preparation for the move and adequate maintenance of treatment during movement requires the skill of experienced medical staff.


The relationship of phonological ability, speech perception, and auditory perception in adults with dyslexia.
This study investigated whether auditory, speech perception, and phonological skills are tightly interrelated or independently contributing to reading. We assessed each of these three skills in 36 adults with a past diagnosis of dyslexia and 54 matched normal reading adults. Phonological skills were tested by the typical threefold tasks, i.e., rapid automatic naming, verbal short-term memory and phonological awareness. Dynamic auditory processing skills were assessed by means of a frequency modulation (FM) and an amplitude rise time (RT); an intensity discrimination task (ID) was included as a non-dynamic control task. Speech perception was assessed by means of sentences and words-in-noise tasks. Group analyses revealed significant group differences in auditory tasks (i.e., RT and ID) and in phonological processing measures, yet no differences were found for speech perception. In addition, performance on RT discrimination correlated with reading but this relation was mediated by phonological processing and not by speech-in-noise. Finally, inspection of the individual scores revealed that the dyslexic readers showed an increased proportion of deviant subjects on the slow-dynamic auditory and phonological tasks, yet each individual dyslexic reader does not display a clear pattern of deficiencies across the processing skills. Although our results support phonological and slow-rate dynamic auditory deficits which relate to literacy, they suggest that at the individual level, problems in reading and writing cannot be explained by the cascading auditory theory. Instead, dyslexic adults seem to vary considerably in the extent to which each of the auditory and phonological factors are expressed and interact with environmental and higher-order cognitive influences.


Factors associated with perinatal mortality in a Brazilian Northeastern capital.
This study investigated factors associated with perinatal mortality in São Luís, Maranhão, Northeastern Brazil. Data on perinatal mortality were obtained from the BRISA birth cohort and from the Mortality Information System, including records of 5,236 births, 70 of which referred to fetal deaths and 36 to early neonatal deaths. Factors associated with mortality were investigated using a hierarchical logistic regression model, resulting in a perinatal mortality coefficient equal to 20.2 per thousand births. Mothers with low education level and without a partner were associated with an increased risk of perinatal death. Moreover, children of mothers who did not have at least six antenatal appointments and with multiple pregnancies (OR= 9.15; 95%CI:4.08-20.53) were more likely to have perinatal death. Perinatal death was also associated with the presence of congenital malformations (OR= 4.13; 95%CI:1.23-13.82), preterm birth (OR= 3.36; 95%CI:1.56-7.22), and low birth weight (OR=11.87; 95%CI:5.46-25.82). In turn, families headed by other family members (OR= 0.29; 95%CI: 0.12 - 0.67) comprised a protective factor for such condition. Thus, the results indicate an association between perinatal mortality and social vulnerability, non-compliance with the recommended number of prenatal appointments, congenital malformations, preterm birth, and low birthweight.


Selenium-dependent and non-selenium-dependent glutathione peroxidase in human tissues of New Zealand residents.
Glutathione peroxidase was assayed in human tissues of New Zealand residents by the coupled assay method. Total glutathione peroxidase was assayed using cumene hydroperoxide. The non-selenium-dependent activity was not detected with t-butyl hydroperoxide and thus was determined from the difference between total activity and the selenium-dependent activity using hydrogen peroxide or t-butyl hydroperoxide. Only selenium-dependent activity was found in whole blood, erythrocytes, platelets and biopsy skeletal muscle. A small non-selenium dependent activity was measured in plasma and a larger activity in biopsy liver supernatant and homogenate. Glutathione-S-transferase was detected in all tissues.


The Amino Acid Transporters of the Glutamate/GABA-Glutamine Cycle and Their Impact on Insulin and Glucagon Secretion.
Intercellular communication is pivotal in optimizing and synchronizing cellular responses to keep homeostasis and to respond adequately to external stimuli. In the central nervous system (CNS), glutamatergic and GABAergic signals are postulated to be dependent on the glutamate/GABA-glutamine cycle for vesicular loading of neurotransmitters, for inactivating the signal and for the replenishment of the neurotransmitters. Islets of Langerhans release the hormones insulin and glucagon, but share similarities with CNS cells in for example transcriptional control of development and differentiation, and chromatin methylation. Interestingly, CNS proteins involved in secretion of the neurotransmitters and emitting their responses as well as the regulation of these processes, are also found in islet cells. Moreover, high levels of glutamate, GABA, and glutamine and their respective vesicular and plasma membrane transporters have been shown in the islet cells and there is emerging support for these amino acids and their transporters playing important roles in the maturation and secretion of insulin and glucagon. In this review, we will discuss the feasibility of recent data in the field in relation to the biophysical properties of the transporters (Slc1, Slc17, Slc32, and Slc38) and physiology of hormone secretion in islets of Langerhans.


Increased free radical production due to subdural hematoma in the rat: effect of increased inspired oxygen fraction.
Acute subdural hematoma (ASDH) complicates about 15%-20% of severe head injury patients and is one of the major causes for bad outcome, yet the pathomechanisms involved are not well understood. This study has employed a recently developed technique to determine whether ASDH induces free radicals in the underlying brain. We also studied the effect of increased inspired oxygen fraction (FiO2) on free radical production, both in the normal rat brain and after ASDH induction. Twelve male Sprague Dawley rats were studied over 5 h (2 h of FiO2 = 30%, 3 h of FiO2 = 100%). Hydroxyl radical production was measured with microdialysis using the salicylate trapping technique by quantitating the 2,3 dihydroxy benzoic acid (2,3 DHBA) and 2,5 dihydroxy benzoic acid (2,5 DHBA), degradation products, in either noninjured brain (n = 6) or after ASDH (n = 6). Both 2,3 DHBA and 2,5 DHBA increased significantly by 39% and 108%, respectively, after the induction of the SDH (p < 0.05). By increasing the FiO2 to 100%, 2 h after ASDH induction, the 2,3 DHBA and 2,5 DHBA further increased only slightly (ns). After increasing the FiO2 to 100% in the noninjured group, the mean level of 2,3 DHBA increased by 56% (p = 0.06, ns). The level of 2,5 DHBA in the dialysate increased significantly by 56% (p < 0.05), when the FiO2 was increased to 100% ASDH results in a significant increase in free radical production. At the same time, prolonged increase in FiO2 does not lead to further increase in free radical production in the injured brain.


Annonacin, a natural mitochondrial complex I inhibitor, causes tau pathology in cultured neurons.
A neurodegenerative tauopathy endemic to the Caribbean island of Guadeloupe has been associated with the consumption of anonaceous plants that contain acetogenins, potent lipophilic inhibitors of complex I of the mitochondrial respiratory chain. To test the hypothesis that annonacin, a prototypical acetogenin, contributes to the etiology of the disease, we investigated whether annonacin affects the cellular distribution of the protein tau. In primary cultures of rat striatal neurons treated for 48 h with annonacin, there was a concentration-dependent decrease in ATP levels, a redistribution of tau from the axons to the cell body, and cell death. Annonacin induced the retrograde transport of mitochondria, some of which had tau attached to their outer membrane. Taxol, a drug that displaces tau from microtubules, prevented the somatic redistribution of both mitochondria and tau but not cell death. Antioxidants, which scavenged the reactive oxygen species produced by complex I inhibition, did not affect either the redistribution of tau or cell death. Both were prevented, however, by forced expression of the NDI1 nicotinamide adenine dinucleotide (NADH)-quinone-oxidoreductase of Saccharomyces cerevisiae, which can restore NADH oxidation in complex I-deficient mammalian cells and stimulation of energy production via anaerobic glycolysis. Consistently, other ATP-depleting neurotoxins (1-methyl-4-phenylpyridinium, 3-nitropropionic, and carbonyl cyanide m-chlorophenylhydrazone) reproduced the somatic redistribution of tau, whereas toxins that did not decrease ATP levels did not cause the redistribution of tau. Therefore, the annonacin-induced ATP depletion causes the retrograde transport of mitochondria to the cell soma and induces changes in the intracellular distribution of tau in a way that shares characteristics with some neurodegenerative diseases.


Screening for frailty in older adults using a self-reported instrument.
OBJECTIVE To validate a screening instrument using self-reported assessment of frailty syndrome in older adults. METHODS This cross-sectional study used data from the Saúde, Bem-estar e Envelhecimento study conducted in Sao Paulo, SP, Southeastern Brazil. The sample consisted of 433 older adult individuals (≥ 75 years) assessed in 2009. The self-reported instrument can be applied to older adults or their proxy respondents and consists of dichotomous questions directly related to each component of the frailty phenotype, which is considered the gold standard model: unintentional weight loss, fatigue, low physical activity, decreased physical strength, and decreased walking speed. The same classification proposed in the phenotype was utilized: not frail (no component identified); pre-frail (presence of one or two components), and frail (presence of three or more components). Because this is a screening instrument, "process of frailty" was included as a category (pre-frail and frail). Cronbach's α was used in psychometric analysis to evaluate the reliability and validity of the criterion, the sensitivity, the specificity, as well as positive and negative predictive values. Factor analysis was used to assess the suitability of the proposed number of components. RESULTS Decreased walking speed and decreased physical strength showed good internal consistency (α = 0.77 and 0.72, respectively); however, low physical activity was less satisfactory (α = 0.63). The sensitivity and specificity for identifying pre-frail individuals were 89.7% and 24.3%, respectively, while those for identifying frail individuals were 63.2% and 71.6%, respectively. In addition, 89.7% of the individuals from both the evaluations were identified in the "process of frailty" category. CONCLUSIONS The self-reported assessment of frailty can identify the syndrome among older adults and can be used as a screening tool. Its advantages include simplicity, rapidity, low cost, and ability to be used by different professionals.


Concurrent Prediction of Dexterous Finger Flexion and Extension Force via Deep Forest.
Neuromuscular injuries can impair hand function and profoundly impacting the quality of life. This has motivated the development of advanced assistive robotic hands. However, the current neural decoder systems are limited in their ability to provide dexterous control of these robotic hands. In this study, we propose a novel method for predicting the extension and flexion force of three individual fingers concurrently using high-density electromyogram (HD-EMG) signals. Our method employs two deep forest models, the flexor decoder and the extensor decoder, to extract relevant representations from the EMG amplitude features. The outputs of the two decoders are integrated through linear regression to predict the forces of the three fingers. The proposed method was evaluated on data from three subjects and the results showed that it consistently outperforms the conventional EMG amplitude-based approach in terms of prediction error and robustness across both target and non-target fingers. This work presents a promising neural decoding approach for intuitive and dexterous control of the fingertip forces of assistive robotic hands.