Development of isotope labeling liquid chromatography mass spectrometry for mouse urine metabolomics: quantitative metabolomic study of transgenic mice related to Alzheimer's disease.
Because of a limited volume of urine that can be collected from a mouse, it is very difficult to apply the common strategy of using multiple analytical techniques to analyze the metabolites to increase the metabolome coverage for mouse urine metabolomics. We report an enabling method based on differential isotope labeling liquid chromatography mass spectrometry (LC-MS) for relative quantification of over 950 putative metabolites using 20 μL of urine as the starting material. The workflow involves aliquoting 10 μL of an individual urine sample for ¹²C-dansylation labeling that target amines and phenols. Another 10 μL of aliquot was taken from each sample to generate a pooled sample that was subjected to ¹³C-dansylation labeling. The ¹²C-labeled individual sample was mixed with an equal volume of the ¹³C-labeled pooled sample. The mixture was then analyzed by LC-MS to generate information on metabolite concentration differences among different individual samples. The interday repeatability for the LC-MS runs was assessed, and the median relative standard deviation over 4 days was 5.0%. This workflow was then applied to a metabolomic biomarker discovery study using urine samples obtained from the TgCRND8 mouse model of early onset familial Alzheimer's disease (FAD) throughout the course of their pathological deposition of beta amyloid (Aβ). It was showed that there was a distinct metabolomic separation between the AD prone mice and the wild type (control) group. As early as 15-17 weeks of age (presymptomatic), metabolomic differences were observed between the two groups, and after the age of 25 weeks the metabolomic alterations became more pronounced. The metabolomic changes at different ages corroborated well with the phenotype changes in this transgenic mice model. Several useful candidate biomarkers including methionine, desaminotyrosine, taurine, N1-acetylspermidine, and 5-hydroxyindoleacetic acid were identified. Some of them were found in previous metabolomics studies in human cerebrospinal fluid or blood samples. This work illustrates the utility of this isotope labeling LC-MS method for biomarker discovery using mouse urine metabolomics.


Dilated Recurrent Neural Networks for Glucose Forecasting in Type 1 Diabetes.
Diabetes is a chronic disease affecting 415 million people worldwide. People with type 1 diabetes mellitus (T1DM) need to self-administer insulin to maintain blood glucose (BG) levels in a normal range, which is usually a very challenging task. Developing a reliable glucose forecasting model would have a profound impact on diabetes management, since it could provide predictive glucose alarms or insulin suspension at low-glucose for hypoglycemia minimisation. Recently, deep learning has shown great potential in healthcare and medical research for diagnosis, forecasting and decision-making. In this work, we introduce a deep learning model based on a dilated recurrent neural network (DRNN) to provide 30-min forecasts of future glucose levels. Using dilation, the DRNN model gains a much larger receptive field in terms of neurons aiming at capturing long-term dependencies. A transfer learning technique is also applied to make use of the data from multiple subjects. The proposed approach outperforms existing glucose forecasting algorithms, including autoregressive models (ARX), support vector regression (SVR) and conventional neural networks for predicting glucose (NNPG) (e.g. RMSE = NNPG, 22.9 mg/dL; SVR, 21.7 mg/dL; ARX, 20.1 mg/dl; DRNN, 18.9 mg/dL on the OhioT1DM dataset). The results suggest that dilated connections can improve glucose forecasting performance efficiently.


A novel method for culturing epithelial cells on a biological substrate.
A device of a new dish for the culturing of cells on a biological substrate, the eye lens capsule, is described. With the aid of this dish it is possible to investigate the possible interrelationships between the cell substratum and various biochemical characteristics of the cell. It is shown that the protein biosynthetic pattern differs between lens cells cultured on lens capsule as a substrate and cells cultured on foil. Moreover, the new dish opens the possibility to culture epithelial cells on a lens capsule, which gives an alternative to the culture of these cells on a collagen substrate.


Potential Predictors of Plasma Fibroblast Growth Factor 23 Concentrations: Cross-Sectional Analysis in the EPIC-Germany Study.
Increased fibroblast growth factor 23 (FGF23), a bone-derived hormone involved in the regulation of phosphate and vitamin D metabolism, has been related to the development of cardiovascular disease (CVD) in chronic kidney disease patients and in the general population. However, what determines higher FGF23 levels is still unclear. Also, little is known about the influence of diet on FGF23. The aim of this study was therefore to identify demographic, clinical and dietary correlates of high FGF23 concentrations in the general population. We performed a cross-sectional analysis within a randomly selected subcohort of the European Prospective Investigation into Cancer and Nutrition (EPIC)-Germany comprising 2134 middle-aged men and women. The Human FGF23 (C-Terminal) ELISA kit was used to measure FGF23 in citrate plasma. Dietary data were obtained at baseline via validated food frequency questionnaires including up to 148 food items. Multivariable adjusted logistic regression showed that men had a 66% lower and smokers a 64% higher probability of having higher FGF23 (≥ 90 RU/mL) levels compared, respectively, with women and nonsmokers. Each doubling in parathyroid hormone, creatinine, and C-reactive protein was related to higher FGF23. Among the dietary factors, each doubling in calcium and total energy intake was related, respectively, to a 1.75 and to a 4.41 fold increased probability of having higher FGF23. Finally, each doubling in the intake of iron was related to an 82% lower probability of having higher FGF23 levels. Results did not substantially change after exclusion of participants with lower kidney function. In middle-aged men and women traditional and non-traditional CVD risk factors were related to higher FGF23 concentrations. These findings may contribute to the understanding of the potential mechanisms linking increased FGF23 to increased CVD risk.


[Removal of a broken silastic catheter with a "snare"-catheter in a neonate].
Silastic central catheters inserted peripherally are used in neonatology since more than 20 years. Typical complications include catheter associated infection, thrombosis and malposition. Pericardial effusion, fracture and embolisation are rare. We report the case of a female newborn with congenital diaphragmatic hernia. A silastic catheter was inserted in the left cubital vein. The catheter ruptured when the obviously occluded lumen was flushed with saline. A 15 cm long catheter fragment located in the right ventricle and pulmonary artery was removed after cardiac catheterization with a "Snare"-catheter.


Avian pineal gland: progonadotropic response in the Japanese quail.
Pinealectomy in juvenile Japanese quail (Coturnix coturnix japonica) exposed to stimulatory photoperiods results in delay of ovarian and oviducal development and decreased pituitary weight. These effects are transitory and occur during a phase of growth immediately preceding the onset of sexual maturity. No effect was found in males. These results suggest that the pineal gland in females has a stimulatory effect on gonadal growth during maturation.


Probiotic bacteria: selective enumeration and survival in dairy foods.
A number of health benefits have been claimed for probiotic bacteria such as Lactobacillus acidophilus, Bifidobacterium spp., and Lactobacillus casei. Because of the potential health benefits, these organisms are increasingly incorporated into dairy foods. However, studies have shown low viability of probiotics in market preparations. In order to assess viability of probiotic bacteria, it is important to have a working method for selective enumeration of these probiotic bacteria. Viability of probiotic bacteria is important in order to provide health benefits. Viability of probiotic bacteria can be improved by appropriate selection of acid and bile resistant strains, use of oxygen impermeable containers, two-step fermentation, micro-encapsulation, stress adaptation, incorporation of micronutrients such as peptides and amino acids and by sonication of yogurt bacteria. This review will cover selective enumeration and survival of probiotic bacteria in dairy foods.


Navigating the fluoroscope's C-arm back into position: an accurate and practicable solution to cut radiation and optimize intraoperative workflow.
During complex image-guided orthopedic trauma procedures, repetitive fluoroscopic scout imaging is performed. A number of preparatory positioning images often must be taken to reproduce a comparable projection. These scout images have no intrinsic clinical relevance but nevertheless expose the patient and the surgical team to considerable radiation, which could be avoided. This study presents and validates a method to decrease intraoperative radiation. Precision, time requirements, and number of scout images for repositioning the fluoroscope, with and without navigation aid, were recorded on 20 test-rig and 3 phantom setups. A commercially available image-guided surgical navigation system (Vector Vision, BrainLAB), originally designed for instrument navigation, was employed to register and retrieve the C-arm positions. A newly developed software computed the necessary moves to reposition the C-arm on an intuitive visual display. Retrieving a given C-arm position with the conventional non-navigated technique required an average of 7 scout images (range, 3 to 12 images). In contrast, navigation-assisted repositioning did not necessitate a single scout image. Deviations from the original projection were minimal for both navigated (0.9 degrees, 95% CI 0.8 to 1.1 degrees) and non-navigated repositioning (0.8 degrees, 95% CI 0.7 to 0.9 degrees). Average positioning times were comparable when navigating the C-arm (46 seconds, 95% CI 41 to 51 seconds) and in scout image-based positioning (49 seconds, 95% CI 44 to 53 seconds). Navigated C-arm positioning avoids multiple scout images and yields sufficient precision for clinical deployment. Radiation exposure can be reduced considerably by a combination of instrument navigation and navigated C-arm positioning.


The impact of cultural background on outcome following traumatic brain injury.
Most traumatic brain injury (TBI) outcome studies have been conducted in developed countries involving individuals from the dominant culture. The present study compared outcomes following TBI in individuals from Culturally and Linguistically Diverse (CALD) backgrounds with those from non-CALD backgrounds. 103 CALD and 103 non-CALD participants with predominantly moderate to severe TBI completed a series of questionnaires an average of 22 months post-injury. Groups were comparable in most demographic and injury-related variables, but CALD participants had lower pre-injury employment rates. Individuals in the CALD group were significantly less independent in light domestic duties, shopping, and financial management and reported significantly lower cognitive independence, mobility, and participation in occupational and social activities than non-CALD participants post-injury. They also reported heightened awareness of post-injury deficits, different beliefs regarding injury consequences and factors aiding recovery, more anxiety and depression symptoms, and less problem-focused coping. Higher functional outcome was associated with having a value system that is Australian, younger age at injury, and higher education. Overall, independent of rehabilitation access, individuals from a CALD background showed poorer functional outcome following TBI than those from a non-CALD background. Addressing this discrepancy should be a priority for rehabilitation programmes.


Heat-stress-induced reproductive failures in chickpea (Cicer arietinum) are associated with impaired sucrose metabolism in leaves and anthers.
Chickpea (Cicer arietinum L.), in its reproductive stage, is sensitive to heat stress (32/20°C or higher as day/night temperatures) with consequent substantial loss of potential yields at high temperatures. The physiological mechanisms associated with reproductive failures have not been established: they constitute the basis of this study. Here, we initially screened a large core-collection of chickpea against heat stress and identified two heat-tolerant (ICC15614, ICCV. 92944) and two heat-sensitive (ICC10685, ICC5912) genotypes. These four genotypes were sown during the normal time of sowing (November-March) and also late (February-April) to expose them to heat stress during reproductive stage (>32/20°C). The genotypes were assessed for damage by heat stress to the leaves and reproductive organs using various indicators of stress injury and reproductive function. In the heat-stressed plants, phenology accelerated as days to flowering and podding, and biomass decreased significantly. The significant reduction in pod set (%) was associated with reduced pollen viability, pollen load, pollen germination (in vivo and in vitro) and stigma receptivity in all four genotypes. Heat stress inhibited pollen function more in the sensitive genotypes than in the tolerant ones, and consequently showed significantly less pod set. Heat stress significantly reduced stomatal conductance, leaf water content, chlorophyll, membrane integrity and photochemical efficiency with a larger effect on heat-sensitive genotypes. Rubisco (carbon-fixing enzyme) along with sucrose phosphate synthase (SPS) and sucrose synthase (SS) (sucrose-synthesising enzymes) decreased significantly in leaves due to heat stress leading to reduced sucrose content. Invertase, a sucrose-cleaving enzyme, was also inhibited along with SPS and SS. The inhibition of these enzymes was significantly greater in the heat-sensitive genotypes. Concurrently, the anthers of these genotypes had significantly less SPS and SS activity and thus, sucrose content. As a result, pollen had considerably lower sucrose levels, resulting in reduced pollen function, impaired fertilisation and poor pod set in heat-sensitive genotypes.


Gas-phase folding of a two-residue model peptide chain: on the importance of an interplay between experiment and theory.
In order to assess the ability of theory to describe properly the dispersive interactions that are ubiquitous in peptide and protein systems, an isolated short peptide chain has been studied using both gas-phase laser spectroscopy and quantum chemistry. The experimentally observed coexistence of an extended form and a folded form in the supersonic expansion was found to result from comparable Gibbs free energies for the two species under the high-temperature conditions (< or = 320 K) resulting from the laser desorption technique used to vaporize the molecules. These data have been compared to results obtained using a series of quantum chemistry methods, including DFT, DFT-D, and post-Hartree-Fock methods, which give rise to a wide range of relative stabilities predicted for these two forms. The experimental observation was best reproduced by an empirically dispersion-corrected functional (B97-D) and a hybrid functional with a significant Hartree-Fock exchange term (M06-2X). In contrast, the popular post-Hartree-Fock method MP2, which is often used for benchmarking these systems, had to be discarded because of a very large basis-set superposition error. The applicability of the atomic counterpoise correction (ACP) is also discussed. This work also introduces the mandatory theoretical examination of experimental abundances. DeltaH(0 K) predictions are clearly not sufficient for discussion of folding, as the conformation inversion temperature is crucial to the conformation determination and requires taking into account thermodynamical corrections (DeltaG) in order to computationally isolate the most stable conformation.


Thoracic response of belted PMHS, the Hybrid III, and the THOR-NT mid-sized male surrogates in low speed, frontal crashes.
Injury to the thorax is the predominant cause of fatalities in crash-involved automobile occupants over the age of 65, and many elderly-occupant automobile fatalities occur in crashes below compliance or consumer information test speeds. As the average age of the automotive population increases, thoracic injury prevention in lower severity crashes will play an increasingly important role in automobile safety. This study presents the results of a series of sled tests to investigate the thoracic deformation, kinematic, and injury responses of belted post mortem human surrogates (PMHS, average age 44 years) and frontal anthropomorphic test devices (ATDs) in low-speed frontal crashes. Nine 29 km/h (three PMHS, three Hybrid III 50th% male ATD, three THOR-NT ATD) and three 38 km/h (one PMHS, two Hybrid III) frontal sled tests were performed to simulate an occupant seated in the right font passenger seat of a mid-sized sedan restrained with a standard (not force-limited) 3-point seatbelt. All occupants were instrumented to record deformation contours and accelerations of the thorax at multiple locations. The ATD subjects were also instrumented to record the internal deformation of the thorax via multi-point tracking systems. For the 29 km/h tests, PMHS maximum chest deflections ranged from 10% to 19% of the undeformed chest depth, and peak mid-spine accelerations ranged from 21 to 24 g. The average peak internal mid-sternal (sternum slider) deflections for the Hybrid III were 23 mm (29 km/h tests) and 30 mm (38 km/h tests). The average maximum Hybrid III sternal deflection of 23 mm measured in the 29 km/h tests corresponds to an AIS 3+ thoracic injury risk of 14% or greater for people 70 years and older. This result suggests that three-point belted elderly occupants without shoulder-belt force limiters could experience non-trivial thoracic injuries in frontal crashes that are below NHTSA's compliance and/or consumer information test severities.


Viral expansion after transfer is a primary driver of influenza A virus transmission bottlenecks.
For many viruses, narrow bottlenecks acting during transmission sharply reduce genetic diversity in a recipient host relative to the donor. Since genetic diversity represents adaptive potential, such losses of diversity are though to limit the opportunity for viral populations to undergo antigenic change and other adaptive processes. Thus, a detailed picture of evolutionary dynamics during transmission is critical to understanding the forces driving viral evolution at an epidemiologic scale. To advance this understanding, we used a novel barcoded virus library and a guinea pig model of transmission to decipher where in the transmission process diversity is lost for influenza A viruses. In inoculated guinea pigs, we show that a high level of viral genetic diversity is maintained across time. Continuity in the barcodes detected furthermore indicates that stochastic effects are not pronounced within inoculated hosts. Importantly, in both aerosol-exposed and direct contact-exposed animals, we observed many barcodes at the earliest time point(s) positive for infectious virus, indicating robust transfer of diversity through the environment. This high viral diversity is short-lived, however, with a sharp decline seen 1-2 days after initiation of infection. Although major losses of diversity at transmission are well described for influenza A virus, our data indicate that events that occur following viral transfer and during the earliest stages of natural infection have a predominant role in this process. This finding suggests that immune selection may have greater opportunity to operate during influenza A transmission than previously recognized.


Global Tuberculosis Report 2020 - Reflections on the Global TB burden, treatment and prevention efforts.
The October 2020 Global TB report reviews TB control strategies and United Nations (UN) targets set in the political declaration at the September 2018 UN General Assembly high-level meeting on TB held in New York. Progress in TB care and prevention has been very slow. In 2019, TB remained the most common cause of death from a single infectious pathogen. Globally, an estimated 10.0 million people developed TB disease in 2019, and there were an estimated 1.2 million TB deaths among HIV-negative people and an additional 208, 000 deaths among people living with HIV. Adults accounted for 88% and children for 12% of people with TB. The WHO regions of South-East Asia (44%), Africa (25%), and the Western Pacific (18%) had the most people with TB. Eight countries accounted for two thirds of the global total: India (26%), Indonesia (8.5%), China (8.4%), the Philippines (6.0%), Pakistan (5.7%), Nigeria (4.4%), Bangladesh (3.6%) and South Africa (3.6%). Only 30% of the 3.5 million five-year target for children treated for TB was met. Major advances have been development of new all oral regimens for MDRTB and new regimens for preventive therapy. In 2020, the COVID-19 pandemic dislodged TB from the top infectious disease cause of mortality globally. Notably, global TB control efforts were not on track even before the advent of the COVID-19 pandemic. Many challenges remain to improve sub-optimal TB treatment and prevention services. Tuberculosis screening and diagnostic test services need to be ramped up. The major drivers of TB remain undernutrition, poverty, diabetes, tobacco smoking, and household air pollution and these need be addressed to achieve the WHO 2035 TB care and prevention targets. National programs need to include interventions for post-tuberculosis holistic wellbeing. From first detection of COVID-19 global coordination and political will with huge financial investments have led to the development of effective vaccines against SARS-CoV2 infection. The world now needs to similarly focus on development of new vaccines for TB utilizing new technological methods.


Intelligence-Dependent Differential Effects of Media Exposure on Executive Function Changes in Children: A Population-Based Cohort Study With a Longitudinal Design.
Excessive media use is known to be associated with executive dysfunction in children, but it's unclear whether this exposure can lead to long-term changes of executive function. This study aimed to investigate the association between media exposure and longitudinal changes in executive function within a population-based study, while considering the potential influence of intelligence. This study used data from 1,209 participants in the Panel Korea Study for Children. The children's media exposure was measured at ages 7 and 8, and executive function was evaluated annually from ages 7 to 10 using the Executive Function Difficulty Screening Questionnaire. Participants were grouped by media exposure level (low, medium, or high), and longitudinal changes in executive function were analyzed using linear mixed effects models. Subgroup analysis was conducted to investigate how executive function changes varied based on intelligence within each media exposure group. Children with high media exposure (n = 97) had severer executive function difficulties than those with low (n = 141) or medium (n = 971) exposure in all waves. The high exposure group demonstrated persistent higher executive function difficulties up to age 10 after controlling for child gender, intelligence, parental education level and maternal depression. Children with intelligence quotient (IQ) ≤ 100 in the medium to high media exposure group had significantly more severe executive function difficulties than those with IQ > 100. This study provided evidence of a longitudinal negative association between media exposure and executive function. The findings suggest that excessive media exposure may lead to long-term changes in executive function in children and highlight the importance of implementing targeted interventions and educational strategies to mitigate the potential negative effects of excessive media use, particularly for children with lower cognitive abilities.


Common forms of childhood esotropia.
To determine the most common forms of childhood esotropia. Prospective, consecutive, observational case series. All esotropic children younger than 11 years of age from a predominantly rural Appalachian region evaluated from August 1, 1995 through July 31, 1998. Demographic and clinical data were collected for all patients. The percentage ratio of the various forms of childhood esotropia. Two hundred twenty-one consecutive children without prior surgical treatment were evaluated for esotropia. One hundred seventeen (52.9%) of the 221 children had some form of accommodative esotropia, 38 (17.2%) were associated with congenital or acquired abnormalities of the central nervous system, 23 (10.4%) displayed acquired nonaccommodative esotropia, 15 (6.8%) resulted from ocular sensory defects, 12 (5.4%) had confirmed congenital esotropia, seven (3.2%) had paralytic esotropia, and an unverified age at onset prevented an accurate categorization in the remaining nine (4.1%). Children with accommodative esotropia accounted for more than half of the study patients and were diagnosed nearly 10 times more frequently than children with congenital esotropia. Esotropic patients with central nervous system defects or with an acquired nonaccommodative deviation were also more common than children with congenital esotropia. Children with congenital esotropia or with a paralytic or sensory cause of their deviation were relatively uncommon.


[Diagnosing depression of a neurotic and subpsychotic character].
Among 176 depressive symptoms informative symptomological combinations were differentiated (by means of mathematical methods) which characterize the neurotic and hypopsychotic depressions and provide a high percent of diagnosis of these conditions by a computer. An additional group of patients was distinguished which occupied a sort of intermediate position between the two mentioned categories. It was found, incidentally, that the vegetative syndromes within the framework of depressions have a rather small informative-diagnostic importance.


Influence of injectate volume on paravertebral spread in erector spinae plane block: An endoscopic and anatomical evaluation.
The paravertebral spread that occurs after erector spinae plane block may be volume-dependent. This cadaveric study was undertaken to compare the extent of paravertebral spread with erector spinae plane block using different dye volumes. After randomization, twelve erector spinae plane blocks were performed bilaterally with either 10 ml or 30 ml of dye at the level of T5 in seven unembalmed cadavers except for two cases of unexpected pleural puncture using the 10 ml injection. Direct visualization of the paravertebral space by endoscopy was performed immediately after the injections. The back regions were also dissected, and dye spread and nerve involvement were investigated. A total of five 10 ml injections and seven 30 ml injections were completed for both endoscopic and anatomical evaluations. No paravertebral spread was observed by endoscopy after any of the 10-ml injections. Dye spread to spinal nerves at the intervertebral foramen was identified by endoscopy at adjacent levels of T5 (median: three levels) in all 30 ml injections. In contrast, the cases with two, four, and three out of five were stained at only the T4, T5, and T6 levels, respectively, with the 10 ml injection. Upon anatomical dissection, all blocks were consistently associated with posterior and lateral spread to back muscles and fascial layers, especially with the 30 ml injections, which showed greater dye expansion. In one 30 ml injection, sympathetic nerve involvement and epidural spread were observed at the level of the injection site. Although paravertebral spread following erector spinae plane block increased in a volume-dependent manner, this increase was variable and not pronounced. As the injectate volume increased for the erector spinae blocks, the injectate spread to the back muscles and fascial layers seemed to be predominantly increased compared with, the extent of paravertebral spread.


On the Mechanism of Bilayer Separation by Extrusion, or Why Your LUVs Are Not Really Unilamellar.
Extrusion through porous filters is a widely used method for preparing biomimetic model membranes. Of primary importance in this approach is the efficient production of single bilayer (unilamellar) vesicles that eliminate the influence of interlamellar interactions and strictly define the bilayer surface area available to external reagents such as proteins. Submicroscopic vesicles produced using extrusion are widely assumed to be unilamellar, and large deviations from this assumption would impact interpretations from many model membrane experiments. Using three probe-free methods-small angle X-ray and neutron scattering and cryogenic electron microscopy-we report unambiguous evidence of extensive multilamellarity in extruded vesicles composed of neutral phosphatidylcholine lipids, including for the common case of neutral lipids dispersed in physiological buffer and extruded through 100-nm diameter pores. In such preparations, only ∼35% of lipids are externally accessible and this fraction is highly dependent on preparation conditions. Charged lipids promote unilamellarity as does decreasing solvent ionic strength, indicating the importance of electrostatic interactions in determining the lamellarity of extruded vesicles. Smaller extrusion pore sizes also robustly increase the fraction of unilamellar vesicles, suggesting a role for membrane bending. Taken together, these observations suggest a mechanistic model for extrusion, wherein the formation of unilamellar vesicles involves competition between bilayer bending and adhesion energies. The findings presented here have wide-ranging implications for the design and interpretation of model membrane studies, especially ensemble-averaged observations relying on the assumption of unilamellarity.


Critical contribution of liver natural killer T cells to a murine model of hepatitis.
Natural killer T (NKT) cells constitute a distinct subpopulation of T cells with a unique antigen specificity, prompt effector functions, and an unusual tissue distribution. NKT cells are especially abundant in the liver, but their physiological function in this organ remains unclear. In the present study, we examined the possible contribution of NKT cells to a murine model of hepatitis induced by i.v. injection of Con A. CD1-deficient mice lacking NKT cells were highly resistant to Con A-induced hepatitis. Adoptive transfer of hepatic NKT cells isolated from wild-type mice, but not from FasL-deficient gld mice, sensitized CD1-deficient mice to Con A-induced hepatitis. Furthermore, adoptive transfer of hepatic mononuclear cells from wild-type mice, but not from CD1-deficient mice, sensitized gld mice to Con A-induced hepatitis. Upon Con A administration, hepatic NKT cells rapidly up-regulated cell surface FasL expression and FasL-mediated cytotoxicity. At the same time, NKT cells underwent apoptosis leading to their rapid disappearance in the liver. These results implicated FasL expression on liver NKT cells in the pathogenesis of Con A-induced hepatitis, suggesting a similar pathogenic role in human liver diseases such as autoimmune hepatitis.


The Internet Intervention Patient Adherence Scale for Guided Internet-Delivered Behavioral Interventions: Development and Psychometric Evaluation.
Patient adherence is defined as the extent to which a patient complies with medical or health advice. At present, there is a lack of reliable and valid measures specifically designed to measure adherence to internet-delivered behavioral interventions. The objective of this study was to develop and psychometrically evaluate a novel measure of adherence to guided internet-delivered behavioral interventions. In collaboration with experienced clinicians and researchers in the field, a 5-item, clinician-rated internet intervention Patient Adherence Scale (iiPAS) was developed. The initial scale was tested in a sample of children and adolescents (N=50) participating in internet-delivered cognitive behavioral therapy (ICBT) studies. A revised version of the iiPAS was then administered to a larger sample of children and adolescents (N=148) with various behavioral problems participating in ICBT trials. The scale was evaluated according to a classical test theory framework. The iiPAS demonstrated excellent internal consistency. Factor analyses revealed one underlying factor, explaining about 80% of the variance, suggesting that the scale captures a homogeneous adherence construct. The iiPAS was strongly associated with objective measures of patient activity in ICBT (number of logins, number of written characters, and completed modules). Furthermore, mid- and posttreatment ratings of the iiPAS were significantly correlated with treatment outcomes. By contrast, objective measures of patient activity in the Web-based platform did not correlate with treatment outcomes. The iiPAS could be a useful tool to measure adherence in a broad range of internet-delivered behavioral interventions.


The Role of Systemic Therapy in Resectable Colorectal Liver Metastases: Systematic Review and Network Meta-Analysis.
Despite multiple randomized trials, the role of perioperative chemotherapy in colorectal cancer liver metastasis (CRLM) is still under debate. In this systematic review and network meta-analysis (NMA), we aim to evaluate the efficacy of perioperative systemic therapies for patients with CRLM. We searched various databases for abstracts and full-text articles published from database inception through May 2021.We included randomized controlled trials (RCTs) comparing the addition of perioperative (post, pre, or both) systemic therapies to surgery alone in patients with CRLM. The outcomes were compared according to the chemotherapy regimen using a random effects model. Outcomes of interest included disease-free survival (DFS) and overall survival (OS). Seven RCTs with a total of 1504 patients with CRLM were included. Six studies included post-operative treatment and one evaluated perioperative (pre- and postoperative) therapy. Fluoropyrimidine-based chemotherapy was the most used systemic therapy. NMA showed benefit of adding perioperative therapy to surgery in terms of DFS (HR 0.73, 95% CI 0.63 to 0.84). However, these findings did not translate into a statistically significant OS benefit (HR 0.88, 95% CI 0.74 to 1.05). NMA did not show any advantage of one regimen over another including oxaliplatin or irinotecan. This systematic review and NMA of 7 RCTs found that the addition of perioperative systemic treatment for resectable CRLM could improve disease-free survival but not overall survival. Based on the findings, addition of perioperative treatment in resectable CRLM should be individualized weighing the risks and benefits.


Supportive therapy for a patient with toxic epidermal necrolysis undergoing plasmapheresis.
A patient with severe toxic epidermal necrolysis underwent 2 cycles of therapeutic plasma exchange and received specialized wound care for widespread skin damage of more than 80% of his body surface area. Extensive involvement of mucous membranes, including the conjunctivas and the oropharyngeal cavity, and damage of his genitourinary organs required meticulous wound care. Daily care of injuries of tissues affected only in the most severe cases of toxic epidermal necrolysis was provided by an experienced intensive care unit nursing team. A meticulous supportive therapy regimen was a major contributing factor to this patient's remission.


Low-Frequency Repetitive Transcranial Magnetic Stimulation of the Right Dorsolateral Prefrontal Cortex Enhances Recognition Memory in Alzheimer's Disease.
The lack of effective pharmacological or behavioral interventions for memory impairments associated with Alzheimer's disease (AD) emphasizes the need for the investigation of approaches based on neuromodulation. This study examined the effects of inhibitory repetitive transcranial magnetic stimulation (rTMS) of prefrontal cortex on recognition memory in AD patients. In a first experiment, 24 mild AD patients received sham and real 1Hz rTMS over the left and right dorsolateral prefrontal cortex (DLPFC), in different sessions, between encoding and retrieval phases of a non-verbal recognition memory task. In a second experiment, another group of 14 AD patients underwent sham controlled repeated sessions of 1Hz rTMS of the right DLPFC across a two week treatment. Non-verbal recognition memory task was performed at baseline, at the end of the two weeks period and at a follow up of 1 month. Right real rTMS significantly improved memory performance compared to right sham rTMS (p = 0.001). Left real rTMS left the memory performance unchanged as compared with left sham rTMS (p = 0.46). The two sham conditions did not differ between each other (p = 0.24). In the second experiment, AD patients treated with real rTMS showed an improvement of memory performance at the end of the two weeks treatment (p = 0.0009), that persisted at 1-month follow-up (p = 0.002). These findings provide evidence that inhibitory rTMS over the right DLPFC can improve recognition memory function in AD patients. They also suggest the importance of a new approach of non-invasive brain stimulation as a promising treatment in AD.


[3H]ouabain binding to cultured rat vascular smooth muscle cells.
The number of Na+ pump units (Bmax) and the equilibrium dissociation constant (Kd) for ouabain as well as parameters of K+ binding to the Na+ pump were examined in in vitro-grown vascular smooth muscle cells ( VSMC ) derived from Sprague-Dawley rats. The technique to measure these variables utilizes analyses of [3H]ouabain displacement from its VSMC receptors by nonlabeled ouabain and K+. The mean values for Bmax and Kd in the cultured VSMCs were 1.95 X 10(5) receptor sites per single VSMC and 2.68 X 10(-6) M, respectively. The equilibrium dissociation constant for K+ (Ki) was 0.92 mM. K+ binding to the cultured VSMCs demonstrated positive cooperativity with a Hill coefficient (n) of 1.78.


Familial hyperparathyroidism.
Twenty-eight members of a family over three generations were studied; six of them had hyperparathyroidism. Four of the six were siblings and one of these had suffered from recurrent hyperparathyroidism. No member of the family showed any other significant endocrine disturbance or active peptic ulceration.Attention is drawn to the prominent lack of symptoms despite dangerous levels of hypercalcaemia and advanced disease with the consequent need for estimation of serum calcium levels in all close relatives. Multiple gland involvement is common in familial hyperparathyroidism and is often coupled with a tendency to recurrence, necessitating long-term follow-up. Histological appearances vary considerably, and the risk of recurrence is not limited to cases showing primary chief cell hyperplasia, with the implication that resection of three parathyroid glands and part of the remaining gland may be the treatment of choice in all cases of familial hyperparathyroidism when multiple gland involvement is found.


Bovine leukemia virus genotype surveillance in cattle at a slaughterhouse in Aichi Prefecture, Japan, in 2019 using polymerase chain reaction combined with restriction fragment length polymorphism.
Polymerase chain reaction (PCR) combined with restriction fragment length polymorphism (RFLP) is commonly used for genotyping bovine leukemia virus (BLV) in slaughterhouses. However, unclassified BLV genotypes have been sporadically reported. To assess the current status of BLV genetic characterization in cattle, PCR-RFLP was performed on blood samples of 170 cattle (84 Japanese Black, 60 Japanese Black x Holstein, and 26 Holstein) from 17 farms (5 prefectures) at a slaughterhouse in Aichi Prefecture in 2019. A total of 65 samples (38.2%) were BLV positive, and genotype 1 was the most predominant (56/65 samples), followed by genotypes 3 (6 samples) and 5 (1 sample), and two unclassified samples. No relationship between the genotypes and breeds was observed. Sequence and phylogenetic analyses demonstrated that unclassified BLV genotypes clustered with genotype 1 sequences were, therefore, not new genotypes.


[Embryotoxicity and the immunodepressive action of tetracycline and its epi- and anhydro- derivatives].
The toxic effect of tetracycline and its epi- and anhydro-derivatives on growing chick embryos and the spleen cells of immunized mice was studied. High acute toxicity of 4-epianhydrotetracycline with respect to the chick embryos was found. Its LD50 was 4.8 times lower than toxicity of tetracycline hydrochloride. The characteristics of the acute toxicity was confirmed by the data on the embryo survival by the time of hatching. The same survival rate, i. e. 12 per cent was observed with the use of tetracycline and 4-epianhydrotetracycline in doses of 1000 and 100gamma per embryo respectively. Comparative investigation of the effect of tetracycline and anhydrotetracycline on the spleen cells revealed high toxicity of anhydrotetracycline which induced the same decrease in the number of the antibody-producing cells as tetracycline when used in doses 40 to 100 times lower than those of tetracycline. High toxicity of the anhydro-derivatives of tetracycline was also observed with respect to their teratogenic effect. Extremely pronounced anomalies in the embryo development were observed after exposure to 500gamma of 4-epianhydrotetracycline.


Artificial light during the polar night disrupts Arctic fish and zooplankton behaviour down to 200 m depth.
For organisms that remain active in one of the last undisturbed and pristine dark environments on the planet-the Arctic Polar Night-the moon, stars and aurora borealis may provide important cues to guide distribution and behaviours, including predator-prey interactions. With a changing climate and increased human activities in the Arctic, such natural light sources will in many places be masked by the much stronger illumination from artificial light. Here we show that normal working-light from a ship may disrupt fish and zooplankton behaviour down to at least 200 m depth across an area of >0.125 km2 around the ship. Both the quantitative and qualitative nature of the disturbance differed between the examined regions. We conclude that biological surveys in the dark from illuminated ships may introduce biases on biological sampling, bioacoustic surveys, and possibly stock assessments of commercial and non-commercial species.


Dosimetry and microdosimetry of targeted radiotherapy.
Dosimetry in targeted radiotherapy (TR) uses different calculation methods, whose degree of refinement is closely conditioned by the particular objective sought. It is more generally performed to establish a correlation between the quantity of radiation delivered to a target and the biological damage observed or that can be reliably predicted. It can thus be used to optimise treatments and allow comparison of different therapeutic approaches, as well as to study the basic methods of irradiation of biological matter. Two broad types of investigations can be found in the literature: microdosimetric ones (stochastic approaches used to study energy deposits) and macrodosimetric ones (non-stochastic or deterministic approaches). The mathematical formalism is consistent between these two types, and the calculation methods currently used are often similar. This review presents different approaches to the dosimetry of radionuclides used in TR. The introduction defines the general problem, the role of dosimetry in TR and the specific problems raised by targeting (non-uniformity of source distributions). The first part considers the types of calculation methods found in TR in relation to the basic quantities used to represent stochastic energy deposit on a cellular scale. In particular, it compares the formalism and the methods used in microdosimetric or conventional macrodosimetric approches. Although microdosimetry, or even track structure calculations, can provide the basic elements for modelling the absorbed dose process, a simplified dosimetric approach may be adequate to describe the phenomena observed. The scheme proposed by the MIRD committee relates to such an approach and is presented together with other methods allowing the calculation of the mean dose delivered (analytic methods, dose point kernels, Monte-Carlo, etc.). The second part shows the application range for the various methods, providing selected examples of dosimetric approaches in TR on different scales, from the organ (or tissues) to the cell or even DNA, and a brief presentation of bone marrow dosimetry.


Epicondylitis: lateral.
Lateral epicondylitis is the most common cause of lateral elbow pain in adults. Although it is typically a self-limiting process, there are many nonsurgical and surgical treatment options available if lateral epicondylitis becomes chronic and continues to cause pain. Its common name, tennis elbow, is somewhat of a misnomer because the condition is often work-related and occurs in athletes and nonathletes alike. Acute onset of symptoms occurs more often in young athletes; chronic, recalcitrant symptoms typically occur in older patients. In this review, we describe the pathogenesis and clinical presentation and the nonsurgical and surgical treatment options currently available.


Thermal lens and heat generation of Nd:YAG lasers operating at 1.064 and 1.34 microm.
We report on a simple and accurate method for determination of thermo-optical and spectroscopic parameters (thermal diffusivity, temperature coefficient of the optical path length change, pump and fluorescence quantum efficiencies, thermal loading, thermal lens focal length, etc) of relevance in the thermal lensing of end-pumped neodymium lasers operating at 1.06- and 1.3- microm channels. The comparison between thermal lensing observed in presence and absence of laser oscillation has been used to elucidate and evaluate the contribution of quantum efficiency and excited sate absorption processes to the thermal loading of Nd:YAG lasers.


Incremental Benefit of Achieving Endoscopic and Histologic Remission in Patients With Ulcerative Colitis: A Systematic Review and Meta-Analysis.
Clinical remission, defined by a composite of patient reported outcomes and Mayo endoscopy subscore (MES) 0 or 1 is a recommended treatment target in patients with ulcerative colitis (UC). We estimated whether incorporating more rigorous remission definitions, of endoscopic remission (MES 0) and histologic remission, affects risk of relapse. Through a systematic review, we identified cohort studies in adults with UC in clinical remission that reported a minimum 12-month risk of clinical relapse, based on MES (0 vs 1) and/or histologic disease activity, in patients with endoscopic remission. Using random effects meta-analysis, we calculated relative and absolute risk of clinical relapse in patients with UC achieving different treatment targets. In a meta-analysis of 17 studies that included 2608 patients with UC in clinical remission, compared to patients achieving MES 1, patients achieving MES 0 had a 52% lower risk of clinical relapse (relative risk, 0.48; 95% CI, 0.37-0.62). The median 12-month risk of clinical relapse in patients with MES 1 was 28.7%; the estimated annual risk of clinical relapse in patients with MES 0 was 13.7% (95% CI, 10.6-17.9). In a meta-analysis of 10 studies in patients in endoscopic remission (MES 0), patients who achieved histologic remission had a 63% lower risk of clinical relapse vs patients with persistent histologic activity (relative risk, 0.37; 95% CI, 0.24-0.56). Estimated annual risk of clinical relapse in who achieved achieving histologic remission was 5.0% (95% CI, 3.3-7.7). In a systematic review and meta-analysis of patients with UC in clinical remission, we observed that patients achieving more rigorous treatment endpoints (endoscopic and histologic remission) have a substantially lower risk of clinical relapse compared with patients achieving clinical remission.


NO-independent regulatory site on soluble guanylate cyclase.
Nitric oxide (NO) is a widespread, potent, biological mediator that has many physiological and pathophysiological roles. Research in the field of NO appears to have followed a straightforward path, and the findings have been progressive: NO and cyclic GMP are involved in vasodilatation; glycerol trinitrate relaxes vascular smooth muscles by bioconversion to NO; mammalian cells synthesize NO; and last, NO mediates vasodilatation by stimulating the soluble guanylate cyclase (sGC), a heterodimeric (alpha/beta) haem protein that converts GTP to cGMP2-4. Here we report the discovery of a regulatory site on sGC. Using photoaffinity labelling, we have identified the cysteine 238 and cysteine 243 region in the alpha1-subunit of sGC as the target for a new type of sGC stimulator. Moreover, we present a pyrazolopyridine, BAY 41-2272, that potently stimulates sGC through this site by a mechanism that is independent of NO. This results in antiplatelet activity, a strong decrease in blood pressure and an increase in survival in a low-NO rat model of hypertension, and as such may offer an approach for treating cardiovascular diseases.


Age-related changes in learning across early childhood: a new imitation task.
Imitation plays a critical role in social and cognitive development, but the social learning mechanisms contributing to the development of imitation are not well understood. We developed a new imitation task designed to examine social learning mechanisms across the early childhood period. The new task involves assembly of abstract-shaped puzzle pieces in an arbitrary sequence on a magnet board. Additionally, we introduce a new scoring system that extends traditional goal-directed imitation scoring to include measures of both children's success at copying gestures (sliding the puzzle pieces) and goals (connecting the puzzle pieces). In Experiment 1, we demonstrated an age-invariant baseline from 1.5 to 3.5 years of age, accompanied by age-related changes in success at copying goals and gestures from a live demonstrator. In Experiment 2, we applied our new task to learning following a video demonstration. Imitation performance in the video demonstration group lagged behind that of the live demonstration group, showing a protracted video deficit effect. Across both experiments, children were more likely to copy gestures at earlier ages, suggesting mimicry, and only later copy both goals and gestures, suggesting imitation. Taken together, the findings suggest that different social learning strategies may predominate in imitation learning dependent upon the degree of object affordance, task novelty, and task complexity.


Fluorescence-enhanced optical imaging in large tissue volumes using a gain-modulated ICCD camera.
A novel image-intensified charge-coupled device (ICCD) imaging system has been developed to perform 3D fluorescence tomographic imaging in the frequency-domain using near-infrared contrast agents. The imager is unique since it (i) employs a large tissue-mimicking phantom, which is shaped and sized to resemble a female breast and part of the extended chest-wall region, and (ii) enables rapid data acquisition in the frequency-domain by using a gain-modulated ICCD camera. Diffusion model predictions are compared to experimental measurements using two different referencing schemes under two different experimental conditions of perfect and imperfect uptake of fluorescent agent into a target. From these experimental measurements, three-dimensional images of fluorescent absorption were reconstructed using a computationally efficient variant of the approximate extended Kalman filter algorithm. The current work represents the first time that 3D fluorescence-enhanced optical tomographic reconstructions have been achieved from experimental measurements of the time-dependent light propagation on a clinically relevant breast-shaped tissue phantom using a gain-modulated ICCD camera.


Human papillomavirus type 94-associated flat warts and review of reported cases.
To date, 10 types of human papillomavirus have been identified that cause flat warts, including human papillomavirus type 3, which belongs to species group 2 of the genus alpha papillomavirus. Among these 10 types, human papillomavirus type 94 is most closely related to human papillomavirus type 10, sharing 86% homology. In this study, we conducted polymerase chain reaction analysis with sequencing on samples obtained from cutaneous lesions located on the face and lower legs of an individual, revealing the presence of human papillomavirus type 94. Dermatoscopic findings revealed numerous dotted vessels within one group of macular brown lesions located on the lower leg, which contributed to the diagnosis of flat warts. An online search revealed that human papillomavirus type 94 has previously been detected in various skin diseases, and we provide a review of prior reports.


The mechanical design of spider silks: from fibroin sequence to mechanical function.
Spiders produce a variety of silks, and the cloning of genes for silk fibroins reveals a clear link between protein sequence and structure-property relationships. The fibroins produced in the spider's major ampullate (MA) gland, which forms the dragline and web frame, contain multiple repeats of motifs that include an 8-10 residue long poly-alanine block and a 24-35 residue long glycine-rich block. When fibroins are spun into fibres, the poly-alanine blocks form (&bgr;)-sheet crystals that crosslink the fibroins into a polymer network with great stiffness, strength and toughness. As illustrated by a comparison of MA silks from Araneus diadematus and Nephila clavipes, variation in fibroin sequence and properties between spider species provides the opportunity to investigate the design of these remarkable biomaterials.


Prognostic value of myeloperoxidase in patients with chest pain.
Inflammation is linked to adverse outcomes in acute coronary syndromes. Myeloperoxidase, an abundant leukocyte enzyme, is elevated in culprit lesions that have fissured or ruptured in patients with sudden death from cardiac causes. Numerous lines of evidence suggest mechanistic links between myeloperoxidase and both inflammation and cardiovascular disease. We assessed the value of plasma levels of myeloperoxidase as a predictor of the risk of cardiovascular events in 604 sequential patients presenting to the emergency department with chest pain. Initial plasma myeloperoxidase levels predicted the risk of myocardial infarction, even in patients who are negative for troponin T (<0.1 ng per milliliter) at base line (P<0.001). Myeloperoxidase levels at presentation also predicted the risk of major adverse cardiac events (myocardial infarction, the need for revascularization, or death) within 30 days and 6 months after presentation (P<0.001). In patients without evidence of myocardial necrosis (defined as those who were negative for troponin T), the base-line myeloperoxidase levels independently predicted the risk of major adverse coronary events at 30 days (unadjusted 2nd, 3rd, and 4th quartile odds ratios, 2.2 [95 percent confidence interval, 1.1 to 4.6], 4.2 [95 percent confidence interval, 2.1 to 8.4], and 4.1 [95 percent confidence interval, 2.0 to 8.4], respectively) and at 6 months. A single initial measurement of plasma myeloperoxidase independently predicts the early risk of myocardial infarction, as well as the risk of major adverse cardiac events in the ensuing 30-day and 6-month periods. Myeloperoxidase levels, in contrast to troponin T, creatine kinase MB isoform, and C-reactive protein levels, identified patients at risk for cardiac events in the absence of myocardial necrosis, highlighting its potential usefulness for risk stratification among patients who present with chest pain.


Identification, characterization and purification to near-homogeneity of a novel 67 kDa phosphotyrosyl protein phosphatase associated with pig lung annexin extract.
During the purification of annexin VI from pig lung, we previously reported the isolation of another 67 kDa protein (protein 67E) differing from the former by immunological reactivity, amino acid composition, inability to interact with anionic phospholipids in the presence of Ca2+ and inability to inhibit phospholipase A2 [Fauvel, Vicendo, Roques, Ragab-Thomas, Granier, Vilgrain, Chambaz, Rochat, Chap & Douste-Blazy (1987) FEBS Lett. 221, 397-402]. Attempts to phosphorylate protein 67E by the protein tyrosine kinase of epidermal-growth-factor receptor revealed a dramatic inhibition of receptor autophosphorylation, which was also observed with insulin receptor. This inhibitory effect was found to be supported by a phosphatase active towards p-nitrophenyl phosphate, phosphotyrosine, [32P]phosphotyrosyl histones and [32P]phosphotyrosyl poly(Glu,Tyr), but inactive towards phosphoserine, phosphothreonine and [32P]phosphoseryl histones. Although not purified to complete homogeneity, the enzyme was purified 273-fold over EGTA extracts from pig lung and corresponded to a monomeric protein displaying an apparent molecular mass of 67 kDa. With [32P]phosphotyrosyl poly(Glu,Tyr) as substrate, the purified enzyme displayed Km and Vmax. values of 10 microM and 1.93 mumol/min per mg respectively, which compare reasonably well with other recently described phosphotyrosyl protein phosphatases. From these data and from its sensitivity to various inhibitors, it is concluded that protein fraction 67E contains a novel phosphotyrosyl protein phosphatase, the association of which with annexin extract might offer a clue to the understanding of its possible targeting to membrane substrates.


How much measurement independence is needed to demonstrate nonlocality?
If nonlocality is to be inferred from a violation of Bell's inequality, an important assumption is that the measurement settings are freely chosen by the observers, or alternatively, that they are random and uncorrelated with the hypothetical local variables. We demonstrate a connection between models that weaken this assumption, allowing partial correlation, and (i) models that allow classical communication between the distant parties, (ii) models that exploit the detection loophole. Even if Bob's choices are completely independent, all correlations from projective measurements on a singlet can be reproduced, with mutual information between Alice's choice and local variables less than or equal to one bit.


The influence model of sponsorship. A Catholic hospital is now part of the "Mayo Clinic".
Saint Marys Hospital was founded in Rochester, MN, in 1889. Constructed by the Sisters of St. Francis, it was staffed by physician members of the local Mayo family. The Mayo practice grew into an association of many physicians and medical residents who later began to staff Rochester Methodist Hospital also; the three healthcare institutions became collectively known as the "Mayo Clinic." By the mid-1980s, billing was so complex for the three still-independent facilities that their leaders decided to integrate more formally. This was done in three phases and resulted in the creation of a single institution known as the Mayo Medical Center. From Saint Marys' standpoint, the facilitating document in this process was a "Sponsorship Agreement" whose purpose was to maintain the sponsor's interests and obligations in the integrated structure. A Sponsorship Board was created to continue the hospital's Catholic tradition, including maintaining its chaplaincy, chapels, religious symbols, and special funds. The Sponsorship Board views the new environment as a special challenge. Its members know that Catholic sponsorship: Comforts patients, who realize they are in the hands of people motivated by the Christian ethic Creates an atmosphere in which patients and their families can seek the spiritual support that often aids healing Strengthens a sense of community among physicians, hospital staff, and administrators The Sponsorship Board hopes the sponsor's influence may come to affect the whole Mayo Medical Center, bringing patients, family members, and staff an "added dimension" of care.


Multimodal Imaging Reveals that Sustained Inhibition of HIF-Prolyl Hydroxylases Induces Opposing Effects on Right and Left Ventricular Function in Healthy Rats.
Hypoxia-inducible factor (HIF) drives transcription of critical hypoxia response genes, increasing the production of red blood cells in low oxygen conditions. In the absence of hypoxia, HIF is degraded by prolyl hydroxylases (HIF-PHs). Pharmacological HIF-PH inhibition stabilizes HIF and is being studied as a treatment for anemia. However, like sustained hypoxia, HIF-PH inhibition may increase pulmonary arterial pressure leading to right ventricular hypertrophy. The aim of this study was to assess the cardiac effects of sustained pharmacological HIF-PH inhibition using multimodal imaging, blood analysis, and histology. Rats were dosed daily with a pan HIF-PH inhibitor or vehicle for 4 weeks followed by a 2-week washout period and underwent longitudinal magnetic resonance imaging (MRI) and echocardiography to simultaneously assess RV and LV function. Blood samples from weeks four and six were analyzed to determine red blood cell composition. Histology was performed on the cardiac tissue from a subset of rats at weeks four and six to assess structural effects. Imaging revealed that RV ejection fraction was reduced in animals receiving HIF-PH inhibitor and resulted in RV hypertrophy. Interestingly, HIF-PH inhibition had the opposite effect on the left ventricle (LV), increasing contractility measured by LV ejection fraction. LV effects were reversed by week six, while RV effects (functional and structural) were sustained. These opposing cardiac effects of HIF-PH inhibition warrant further study to both understand the potential negative effects on RV structure and function and investigate the therapeutic potential of increased LV contractility for conditions like heart failure.


Phosphate-regulated expression of the SARS-CoV-2 receptor-binding domain in the diatom Phaeodactylum tricornutum for pandemic diagnostics.
The worldwide COVID-19 pandemic caused by the SARS-CoV-2 betacoronavirus has highlighted the need for a synthetic biology approach to create reliable and scalable sources of viral antigen for uses in diagnostics, therapeutics and basic biomedical research. Here, we adapt plasmid-based systems in the eukaryotic microalgae Phaeodactylum tricornutum to develop an inducible overexpression system for SARS-CoV-2 proteins. Limiting phosphate and iron in growth media induced expression of the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein from the P. tricornutum HASP1 promoter in the wild-type strain and in a histidine auxotrophic strain that alleviates the requirement for antibiotic selection of expression plasmids. The RBD was purified from whole cell extracts (algae-RBD) with yield compromised by the finding that 90-95% of expressed RBD lacked the genetically encoded C-terminal 6X-histidine tag. Constructs that lacked the TEV protease site between the RBD and C-terminal 6X-histidine tag retained the tag, increasing yield. Purified algae-RBD was found to be N-linked glycosylated by treatment with endoglycosidases, was cross-reactive with anti-RBD polyclonal antibodies, and inhibited binding of recombinant RBD purified from mammalian cell lines to the human ACE2 receptor. We also show that the algae-RBD can be used in a lateral flow assay device to detect SARS-CoV-2 specific IgG antibodies from donor serum at sensitivity equivalent to assays performed with RBD made in mammalian cell lines. Our study shows that P. tricornutum is a scalable system with minimal biocontainment requirements for the inducible production of SARS-CoV-2 or other coronavirus antigens for pandemic diagnostics.


Three-dimensional loading model for periodontal ligament regeneration in vitro.
In this study we present a new three-dimensional (3D) model to study effects of mechanical loading on tendon/ligament formation in vitro. The model mimics a functional periodontal ligament (PDL), which anchors dental roots to the jaw bone and transfers the axial load of mastication to the jaw bone. A collagen gel containing human PDL fibroblasts was seeded in a PDL space between an artificial root and bone surface. The effects of 3-day loading on the fibroblasts were studied in vitro by axial and intermittent displacement of the root to which the gel was attached. Cell responses were recorded by measuring expression of three sets of genes: (i) cyclooxygenase 1 and 2 (COX-1, COX-2) producing prostaglandins (signaling molecules); (ii) Runx2, a transcription factor for the osteogenic lineage; and (iii) the extracellular matrix proteins osteopontin, dentin matrix protein 1, and collagen type I (COL1). Loading for 3 days resulted in magnitude-dependent changes in the expression of COX-2 and COL1. A low loading magnitude significantly decreased COX-2 expression, an intermediate magnitude increased its expression, while a high magnitude increased COL1 expression. We concluded that the 3D loading model provides a useful, well-controlled method to examine ligament fibroblast responses to mechanical loading. The model may serve to explore the application of mechanical loading as an anabolic factor for ligament reconstruction.


Recapitulation of germ cell- and pituitary-specific expression with 1.6 kb of the cystatin-related epididymal spermatogenic (Cres) gene promoter in transgenic mice.
The Cres (cystatin-related epididymal spermatogenic) gene encodes the defining member of a new subgroup within the family 2 cystatins of cysteine protease inhibitors. Cres expression is highly tissue- and cell-specific, with messenger RNA (mRNA) present in the testicular round/elongating spermatids, proximal caput epididymal epithelium, gonadotroph cells in the anterior pituitary gland, and corpus luteum of the ovary. To begin to elucidate the molecular mechanisms controlling the tissue- and cell-specific expression of the Cres gene, transgenic mice were generated containing 1.6 kilobases (kb) of the mouse Cres promoter linked to the bacterial chloramphenicol acetyltransferase (CAT) reporter gene. A CAT enzyme-linked immunosorbent assay detected CAT protein in the testis, epididymis, isolated cauda epididymal spermatozoa, and anterior pituitary gland from mice heterozygous and homozygous for the transgene. However, reverse transcription (RT)-PCR did not detect CAT mRNA in any regions of the epididymis, suggesting that the CAT protein detected in the epididymis was from spermatozoa. RT-PCR also did not detect CAT mRNA in the ovary. RT-PCR analysis of the testes from mice of different postnatal ages showed CAT mRNA first detected at day 22, which correlated with the first appearance of Cres mRNA and with the presence of round spermatids. These studies demonstrate that 1.6 kb of Cres promoter contains the DNA elements necessary for germ cell and pituitary gland-specific expression but lacks critical sequences necessary for expression in the epididymis and ovary.


SIRT2 directs the replication stress response through CDK9 deacetylation.
Sirtuin 2 (SIRT2) is a sirtuin family deacetylase that directs acetylome signaling, protects genome integrity, and is a murine tumor suppressor. We show that SIRT2 directs replication stress responses by regulating the activity of cyclin-dependent kinase 9 (CDK9), a protein required for recovery from replication arrest. SIRT2 deficiency results in replication stress sensitivity, impairment in recovery from replication arrest, spontaneous accumulation of replication protein A to foci and chromatin, and a G2/M checkpoint deficit. SIRT2 interacts with and deacetylates CDK9 at lysine 48 in response to replication stress in a manner that is partially dependent on ataxia telangiectasia and Rad3 related (ATR) but not cyclin T or K, thereby stimulating CDK9 kinase activity and promoting recovery from replication arrest. Moreover, wild-type, but not acetylated CDK9, alleviates the replication stress response impairment of SIRT2 deficiency. Collectively, our results define a function for SIRT2 in regulating checkpoint pathways that respond to replication stress through deacetylation of CDK9, providing insight into how SIRT2 maintains genome integrity and a unique mechanism by which SIRT2 may function, at least in part, as a tumor suppressor protein.


Levodopa Is a Double-Edged Sword for Balance and Gait in People With Parkinson's Disease.
The effects of levodopa on balance and gait function in people with Parkinson's disease (PD) is controversial. This study compared the relative responsiveness to l-dopa on six domains of balance and gait: postural sway in stance; gait pace; dynamic stability; gait initiation; arm swing; and turning in people with mild and severe PD, with and without dyskinesia. We studied 104 subjects with idiopathic PD (H & Y II [n = 52] and III-IV [n = 52]) and 64 age-matched controls. Subjects performed a mobility task in the practical off state and on l-dopa: standing quietly for 30 seconds, initiating gait, walking 7 meters, and turning 180 degrees. Thirty-four measures of mobility were computed from inertial sensors. Standardized response means were used to determine relative responsiveness to l-dopa. The largest improvements with l-dopa were found for arm swing and pace-related gait measures. Gait dynamic stability was unaffected by PD and not responsive to l-dopa. l-dopa reduced turning duration, but only in subjects with severe PD. In contrast to gait, postural sway in quiet standing increased with l-dopa, especially in the more severely affected subjects. The increase in postural sway, as well as decrease in turning duration and exaggerated arm swing with l-dopa was observed only for subjects with dyskinesia at the time of testing. The observed spectrum of l-dopa responsiveness in balance and gait measures suggests that multiple neural circuits control balance and gait. Many of the negative effects of l-dopa may be directly or indirectly caused by dyskinesia.


Photosensitive surfactants: micellization and interaction with DNA.
Recently, photosensitive surfactants have re-attracted considerable attention. It has been shown that their association with oppositely charged biologically important polyelectrolytes, such as DNA or microgels, can be efficiently manipulated simply by light exposure. In this article, we investigate the self-assembly of photosensitive surfactants as well as their interactions with DNA by calorimetric and spectroscopic methods. Critical micelle concentration (CMC), standard micellization enthalpy, entropy, and Gibbs energy were determined in different conditions (ionic strengths and temperatures) for a series of cationic surfactants with an azobenzene group in their tail. It is shown, that aggregation forces of photosensitive units play an important role in the micellization giving the major contribution to the micellization enthalpy. The onset of the aggregation can be traced from shift of the absorption peak position in the UV-visible spectrum. Titration UV-visible spectroscopy is used as an alternative, simple, and sensitive approach to estimate CMC. The titration UV-visible spectroscopy was also employed to investigate interactions (CAC: critical aggregation concentration, precipitation, and colloidal stabilization) in the DNA-surfactant complex.


Evidence-Based Message Strategies to Increase Public Support for State Investment in Early Childhood Education: Results from a Longitudinal Panel Experiment.
Policy Points Investments in early childhood education can have long-lasting influence on health and well-being at later stages of the life course. Widespread public support and strategies to counter opposition will be critical to the future political feasibility of enhancing early childhood policies and programs. Simple advocacy messages emphasizing the need for affordable, accessible, high-quality childcare for all can increase public support for state investments in these policies. Policy narratives (short stories with a setting, characters, and a plot that unfolds over time and offers a policy solution to a social problem) that describe structural barriers to childcare and policy solutions to address these barriers may be particularly effective to persuade individuals inclined to oppose such policies to become supportive. Inoculation messages (messages designed to prepare audiences for encountering and building resistance to opposing messages) may protect favorable childcare policy attitudes in the face of oppositional messaging. Early childhood education (ECE) programs enhance the health and social well-being of children and families. This preregistered, randomized, controlled study tested the effectiveness of communication strategies to increase public support for state investments in affordable, accessible, and high-quality childcare for all. At time 1 (August-September 2019), we randomly assigned members of an online research panel (n = 4,363) to read one of four messages promoting state investment in childcare policies and programs, or to a no-exposure control group. Messages included an argument-based message ("simple pro-policy"), a message preparing audiences for encountering and building resistance to opposing messages ("inoculation"), a story illustrating the structural nature of the problem and solution ("narrative"), and both inoculation and narrative messages ("combined"). At time 2 (two weeks later) a subset of respondents (n = 1,436) read an oppositional anti-policy message and, in two conditions, another narrative or inoculation message. Ordinary least squares regression compared groups' levels of support for state investment in childcare policies and programs. As hypothesized, respondents who read the narrative message had higher support for state investment in childcare policies than those who read the inoculation message or those in the no-exposure control group at time 1. Among respondents who were initially opposed to such investments, those who read the narrative had greater support than respondents who read the simple pro-policy message. Those who received the inoculation message at time 2 were more resistant to the anti-policy message than respondents who did not receive such a message, but effects from exposures to strategic messages at time 1 did not persist at follow-up. Results offer guidance for policy advocates seeking to increase public support for early childhood policies and programs and could inform broader efforts to promote high-value policies with potential to improve population health.


Acute 'strokenomics': efficacy and economic analyses of alteplase for acute ischemic stroke.
Cerebrovascular disease is the leading cause of adult disability in Western countries and, as such, is associated with a considerable and increasing economic burden. Prevention of disability is preferable to a costly and prolonged period of rehabilitation; however, in this regard, we have only a limited number of acute treatments. Treatment of ischemic stroke with the tissue plasminogen activator alteplase has a considerable and compelling evidence base confirming its clinical efficacy. Good-quality trial data are complemented by years of 'real-life' clinical experience. There is limited pharmacoeconomic literature describing the use of alteplase in ischemic stroke, and published data are consistent and broadly supportive of its use in select patient populations. Delivery of stroke and other medical care can differ across countries; however, models based on American, Canadian and European data suggest that following initial monetary investment, net healthcare-associated costs for tissue plasminogen activator-treated patients are less than for untreated contemporaries. Potential savings are driven by reduced disability and associated long-term care costs. At present, robust economic data are only available for the period of 1-year following stroke. Continuing cost savings associated with reduced disability in the longer term make intuitive sense but are yet to be confirmed. A recurring theme in these economic analyses is of reducing healthcare costs associated with the increasing use of thrombolysis. In this regard, it is unfortunate that rates of tissue plasminogen activator utilization remain modest in most centers. This review will discuss the economics of alteplase in acute stroke, making particular reference to the current and projected economic burden of stroke, the evidence base for thrombolysis and published literature on the economics analyses of this therapy.


Atrial natriuretic factor reduces renin release by opposing alpha-adrenoceptor activity.
To examine how atrial natriuretic factor (ANF) inhibits renin release during renal sympathetic nerve stimulation, experiments were performed in barbiturate-anesthetized dogs. In five dogs, intravenous ANF infusion (50 ng.min-1.kg body wt-1) reduced renin release induced by renal nerve stimulation (1 Hz) from 16.8 +/- 8.4 to 3.5 +/- 2.1 micrograms angiotensin I (ANG I)/min. In two groups, renin release was raised by ureteral occlusion, which enhances the effects of beta-adrenoceptor stimulation and increased prostaglandin synthesis. During ureteral occlusion, intrarenal infusion of isoproterenol (0.2 micrograms.min-1.kg body wt-1) increased renin release in eight dogs to 82.6 +/- 10.9 micrograms ANG I/min, which was not significantly reduced by ANF infusion (81.1 +/- 10.1 micrograms ANG I/min). Similarly, intrarenal infusion of arachidonic acid (80 micrograms.min-1.kg body wt-1) during ureteral occlusion increased renin release in five dogs to 22.2 +/- 3.0 micrograms ANG I/min, which was not significantly reduced by ANF infusion (22.5 +/- 3.5 micrograms ANG I/min). Finally, in six dogs examined at free urine flow, intrarenal infusion of phenylephrine, an alpha-adrenergic agonist, raised renin release from 0.5 +/- 0.3 to 20.1 +/- 6.8 micrograms ANG I/min, which was reduced to 10.6 +/- 3.9 micrograms ANG I/min by intravenous ANF infusion (100 ng.min-1.kg body wt-1). These results indicate that ANF does not counteract stimulation of renin release by beta-adrenoceptors and prostaglandins but reduces nerve-stimulated renin release by opposing alpha-adrenoceptor activity.


[Nursing theories--friends or foes? Student nurses' perspective].
Nursing researchers consider nursing theories too theoretical and complicated to apply in practice. In this study student nurses in their last term are being interviewed about their ideas of nursing theories. A lot of different views emerged. The conclusion to make from this study is that a frequent presentation both practical and theoretical is required in order to make theories more accessible to nursing students. It is also important that different kinds of theories are exposed to give the students the possibility to find the theory most suited for the personality. This also gives the student the advantage to choose the essence from each theory. This is a way to establish a natural relationship between students and nursing theories and thereby the students will find easier ways to apply nursing theories in different nursing actions.


Epidermal patterning and stomatal development in Gnetales.
The gymnosperm order Gnetales, which has contentious phylogenetic affinities, includes three extant genera (Ephedra, Gnetum, Welwitschia) that are morphologically highly divergent and have contrasting ecological preferences: Gnetum occupies mesic tropical habitats, whereas Ephedra and Welwitschia occur in arid environments. Leaves are highly reduced in Ephedra, petiolate with a broad lamina in Gnetum and persistent and strap-like in Welwitschia. We investigate stomatal development and prepatterning stages in Gnetales, to evaluate the substantial differences among the three genera and compare them with other seed plants. Photosynthetic organs of representative species were examined using light microscopy, scanning electron microscopy and transmission electron microscopy. Stomata of all three genera possess lateral subsidiary cells (LSCs). LSCs of Ephedra are perigene cells derived from cell files adjacent to the stomatal meristemoids. In contrast, LSCs of Gnetum and Welwitschia are mesogene cells derived from the stomatal meristemoids; each meristemoid undergoes two mitoses to form a 'developmental triad', of which the central cell is the guard mother cell and the lateral pair are LSCs. Epidermal prepatterning in Gnetum undergoes a 'quartet' phase, in contrast with the linear development of Welwitschia. Quartet prepatterning in Gnetum resembles that of some angiosperms but they differ in later development. Several factors underpin the profound and heritable differences observed among the three genera of Gnetales. Stomatal development in Ephedra differs significantly from that of Gnetum and Welwitschia, more closely resembling that of other extant gymnosperms. Differences in epidermal prepatterning broadly reflect differences in growth habit between the three genera.


[Health education: a historical reflection of its practices].
This text aims to contextualize the transformations occurred in the practice of health education since the political scenario in the end of 19th century, when the first extended initiatives of the Brazilian State in the health field were organized, until the creation of the Brazilian Unified Health System.


Structure of the SHOC2-MRAS-PP1C complex provides insights into RAF activation and Noonan syndrome.
SHOC2 acts as a strong synthetic lethal interactor with MEK inhibitors in multiple KRAS cancer cell lines. SHOC2 forms a heterotrimeric complex with MRAS and PP1C that is essential for regulating RAF and MAPK-pathway activation by dephosphorylating a specific phosphoserine on RAF kinases. Here we present the high-resolution crystal structure of the SHOC2-MRAS-PP1C (SMP) complex and apo-SHOC2. Our structures reveal that SHOC2, MRAS, and PP1C form a stable ternary complex in which all three proteins synergistically interact with each other. Our results show that dephosphorylation of RAF substrates by PP1C is enhanced upon interacting with SHOC2 and MRAS. The SMP complex forms only when MRAS is in an active state and is dependent on SHOC2 functioning as a scaffolding protein in the complex by bringing PP1C and MRAS together. Our results provide structural insights into the role of the SMP complex in RAF activation and how mutations found in Noonan syndrome enhance complex formation, and reveal new avenues for therapeutic interventions.


Study on the molecular mechanism of Rac3 on regulating autophagy in human lung cancer cells.
Rac3 plays an important role in regulating tumorigenesis. Autophagy plays a vital role in tumorigenesis and tumor progression. The relationship between the two remains unclear. The objective of the present study was to determine the specific molecular mechanism of intracellular Rac3 in regulating autophagy and reveal the relationship between tumor cell autophagy and apoptosis. A laser confocal microscope was used to photograph the accumulated EGFP-MAP1LC3 spots for investigating the relationship between Rac3 and autophagy at the cellular level. Immunoblotting was also used to investigate the relationship between Rac3 and autophagy. The autophagy flux arising from inhibition of Rac3 was detected with autophagy inhibitors and ATG5 and ATG7 siRNA interference experiments. ATF4 and DDIT4 siRNA interference and overexpression experiments were conducted to investigate the relationship between endoplasmic reticulum stress, the MTOR signaling pathway, and autophagy arising from inhibition of Rac3. Co-immunoprecipitation experiments were performed to investigate the interaction between Rac3 and proteins related to endoplasmic reticulum stress. Co-immunoprecipitation was performed to investigate the structural domains between Rac3 and HSPA5. The expression of ATF4 and DDIT4 was upregulated, which inhibited the MTOR signaling pathway and induced autophagy of human non-small cell lung cancer cells after Rac3 siRNA was introduced. The degree of acetylation of the substrate, HSPA5, increased and the endoplasmic reticulum stress response was activated after Rac3 was inhibited. In conclusion, the degree of acetylation of HSPA5 increased and it was dissociated from the receptor, EIF2AK3, on the endoplasmic reticulum membrane, thus causing the endoplasmic reticulum stress response. Endoplasmic reticulum stress activated the expression of the ATF4 protein, upregulated the level of DDIT4, inhibited the MTOR signaling pathway, and caused cellular autophagy.


A response to yet another defence of ECT in the absence of robust efficacy and safety evidence.
It is estimated that electroconvulsive therapy is still administered to approximately a million people a year. It involves passing enough electric current through the human brain, eight to twelve times, to cause convulsions, in the hope of somehow alleviating emotional suffering, primarily depression. There have only ever been 11 placebo-controlled studies (where general anaesthesia is administered but the electric shock is withheld), all of which were pre-1986, had very small sample sizes and were seriously methodologically flawed. Five of these studies found no difference between the two groups at the end of treatment, four found ECT produced better outcomes for some patients, and two produced mixed results, including one where psychiatrists' ratings produced a difference, but the ratings of nurses and patients did not. In the 80 years since the first ECT no studies have found any evidence that ECT is better than placebo beyond the end of treatment. Nevertheless, all five meta-analyses relying on these studies have somehow concluded that ECT is more effective than placebo despite the studies' multiple failings. Meanwhile, evidence of persistent or permanent memory loss in 12% to 55% of patients has accumulated. Attempts to highlight this failure of ECT proponents to provide robust evidence that their treatment is effective and safe are routinely dismissed, diminished, denied and denounced. This paper responds to one such attempt, by Drs Meechan, Laws, Young, McLoughlin and Jauhar, to discredit two systematic reviews of the eleven pre-1986 studies, in 2010 and 2019, the latter of which also reviewed five meta-analyses that had ignored the studies' failings. The criticisms and claims of the recent crtiique of the two systematic reviews are examined in detail, by the first author of both reviews, for accuracy, relevance and logic. The critique is found to include multiple errors, misrepresentations, omissions, inconsistencies and logical flaws. It is concluded that Meechan et al. fail to make a fact-based, coherent argument against suspending ECT pending a series of large, carefully designed placebo-controlled studies to establish whether ECT does have any beneficial effects against which to weigh the significant established adverse effects.


Collaborative goal setting with adults attending physiotherapy at a specialist neuromuscular centre: is it always appropriate? A cross-sectional survey.
Collaborative goal setting is an integral component of treatment planning for adults with neuromuscular disorders (NMD). However, due to the unique challenges for these individuals, identifying a process for goal setting that is advantageous for all can be problematic. This study aimed to evaluate collaborative goal setting at a specialist NMD centre, as reported by service users attending physiotherapy. It also aimed to generate discussion about collaborative goal setting and the practice of goal setting in adults with NMD in order to inform future practice. Specialist NMD community-based centre in the UK. One hundred and four adults with NMD who attended the centre. Cross-sectional survey. Thematic and content analyses of goals set were performed alongside demographic data collection. One hundred and four patients (34 females) with a range of neuromuscular conditions - including Becker, facioscapularhumeral, limb girdle, Duchenne and myotonic muscular dystrophies - completed the survey. Thirty-six respondents (37%) stated that they had set goals with the physiotherapist, whilst 62 (63%) stated that they had not set goals with the physiotherapist. Respondents' goals were grouped into four themes: symptom management, maintenance, improving physical condition, and learning to live with the condition. Readiness to take part in collaborative goal setting is unique to each individual. Physiotherapists need to be skilful in supporting adults with NMD through the goal-setting process until they are capable of sharing responsibility. Setting personal goals to improve emotional well-being may help to develop confidence to take more control of their situation, hence facilitating skills in self-management.


Policymaker, health provider and community perspectives on male involvement during pregnancy in southern Mozambique: a qualitative study.
Increasing male involvement during pregnancy is considered an important, but often overlooked intervention for improving maternal health in sub-Saharan Africa. Intervention studies aimed at improving maternal health mostly target mothers hereby ignoring the crucial role their partners play in their ability to access antenatal care (ANC) and to prevent and treat infectious diseases like HIV and malaria. Very little is known about the current level of male involvement and barriers at different levels. This study explores the attitudes and beliefs of health policymakers, health care providers and local communities regarding men's involvement in maternal health in southern Mozambique. Ten key informant interviews with stakeholders were carried out to assess their attitudes and perspectives regarding male involvement in programmes addressing maternal health, followed by 11 days of semi structured observations in health care centers. Subsequently 16 focus group discussions were conducted in the community and at provider level, followed by three in depth couple interviews. Analysis was done by applying a socio-ecological systems theory in thematic analysis. Results show a lack of strategy and coherence at policy level to stimulate male involvement in maternal health programmes. Invitation cards for men are used as an isolated intervention in health facilities but these have not lead to the expected success. Providers have a rather passive attitude towards male involvement initiatives. In the community however, male attendance at ANC is considered important and men are willing to take a more participating role. Main barriers are the association of male attendance at ANC with being HIV infected and strong social norms and gender roles. On the one hand men are seen as caretakers of the family by providing money and making the decisions. On the other hand, men supporting their wife by showing interest in their health or sharing household tasks are seen as weak or as a manifestation of HIV seropositivity. A clear strategy at policy level and a multi-level approach is needed. Gender-equitable relationships between men and women should be encouraged in all maternal health interventions and providers should be trained to involve men in ANC.


Gabapentin Does Not Appear to Improve Postoperative Pain and Sleep Patterns in Patients Who Concomitantly Receive Regional Anesthesia for Lower Extremity Orthopedic Surgery: A Randomized Control Trial.
In recent years, gabapentin has gained popularity as an adjuvant therapy for the treatment of postoperative pain. Numerous studies have shown a decrease in pain score, even with immediate postoperative activity, which is significant for early post-op ambulation and regaining functionality sooner. However, studies have been in conclusive in patients undergoing lower extremity orthopedic surgery. For this reason, we hoped to study the effect of gabapentin on postoperative pain in patients undergoing total knee arthroplasty, total hip arthroplasty, or a hip fracture repair. This was done in the setting of ensuring adequate postoperative analgesia with regional blocks and opioid PCA, as is protocol at our institution. Given the sedative effects of gabapentin and the potential for improving postoperative sleep patterns, we also studied the drug's effect on this aspect of our patient's postoperative course. We utilized the Pittsburg Sleep Quality Index and Visual Analog Scale for pain to obtain a more objective standardized score amongst our study population. Our results indicate that gabapentin does not offer any additional relief in pain or improve sleep habits in patients who have received either a femoral or lumbar plexus block for lower extremity orthopedic surgery. This trial is registered with NCT01546857.


Dose-injury relationships for cryoprotective agent injury to human chondrocytes.
Vitrification of articular cartilage (AC) could enhance tissue availability but requires high concentrations of cyroprotective agents (CPAs). This study investigated relative injuries caused by commonly used CPAs. We hypothesized that the in situ chondrocyte dose-injury relationships of five commonly used CPAs are nonlinear and that relative injuries could be determined by comparing cell death after exposure at increasing concentrations. Human AC samples were used from four patients undergoing total knee arthroplasty surgery. Seventy μm slices were exposed in a stepwise protocol to increasing concentrations of 5 CPAs (max = 8 M); dimethyl sulfoxide (Me(2)SO), glycerol (Gly), propylene glycol (PG), ethylene glycol (EG), and formamide (FM). Chondrocyte viability was determined by membrane integrity stains. Statistical analysis included t-tests and nonlinear least squares estimation methods. The dose-injury to chondrocytes relationships for all CPAs were found to be nonlinear (sigmoidal best fit). For the particular loading protocol in this study, the data identified the following CPA concentrations at which chondrocyte recoveries statistically deviated significantly from the control recovery; 1 M for Gly, 4 M for FM and PG, 6 M for Me(2)SO, and 7 M for EG. Comparison of individual means demonstrated that Gly exposure resulted in the lowest recovery, followed by PG, and then Me(2)SO, FM and EG in no specific order. The information from this study provides an order of damage to human chondrocytes in situ of commonly used CPAs for vitrification of AC and identifies threshold CPA concentrations for a stepwise loading protocol at which chondrocyte recovery is significantly decreased. In general, Gly and PG were the most damaging while DMSO and EG were among the least damaging.


Solvation response along the reaction coordinate in the active site of ketosteroid isomerase.
A light-activated reaction analog has been developed to mimic the catalytic reaction cycle of Delta(5)-3-ketosteroid isomerase to probe the functionally relevant protein solvation response to the catalytic charge transfer. Delta(5)-3-ketosteroid isomerase from Pseudomonas putida catalyzes a C-H bond cleavage and formation through an enolate intermediate. Conversion of the ketone substrate to the enolate intermediate is simulated by a photoacid bound to the active site oxyanion hole. In the ground state, the photoacid electrostatically resembles the enolate intermediate while the low pK(a) excited state resembles the ketone starting material. Time-resolved fluorescence experiments with photoacids coumarin 183 and equilenin show the active site of Delta(5)-3-ketosteroid isomerase to be largely unperturbed by the light-activated reaction. The small solvation response for the photoacid at the active site as compared with a simple solvent suggests the active site does not significantly change its electrostatic environment during the catalytic cycle. Instead, the reaction takes place in an electrostatically preorganized environment.


Scavenging systems for reactive carbonyls in the cyanobacterium Synechocystis sp. PCC 6803.
To elucidate the scavenging systems of sugar- and lipid-derived reactive carbonyls (RCs) in the cyanobacterium Synechocystis sp. PCC 6803 (S. 6803), we selected proteins from S. 6803 based on amino-acid (AA) sequence similarities with proteins from Arabidopsis thaliana, and characterized the properties of the GST-fusion proteins expressed. Slr0942 catalyzed the aldo-keto reductase (AKR) reaction scavenging mainly sugar-derived RCs, methylglyoxal (MG). Slr1192 is the medium-chain dehydrogenase/redutase (MDR). It catalyzed the AKR reaction scavenging several lipid-derived RCs, acrolein, propionaldehyde, and crotonaldehyde. Slr0315 is a short-chain dehydrogenase/redutase (SDR), and it catalyzed only the reduction of MG in the AKR reaction. Slr0381 catalyzed the conversion of hemithioacetal to S-lactoylglutahione (SLG) in the glyoxalase (GLX) 1 reaction. Sll1019 catalyzed the conversion of SLG to glutathione and lactate in the GLX2 reaction. GLX1 and GLX2 compose the glyoxalase system, which scavenges MG. These enzymes contribute to scavenging sugar- and lipid-derived RCs as scavenging systems.