Sequence of the sites phosphorylated by protein kinase C in the smooth muscle myosin light chain.
We have determined the sequence of the sites phosphorylated by protein kinase C in the turkey gizzard smooth muscle myosin light chain. In contrast to previous work (Nishikawa, M., Hidaka, H., and Adelstein, R. S. (1983) J. Biol. Chem. 258, 14069-14072), two-dimensional tryptic peptide maps of both heavy meromyosin and the isolated myosin light chain showed two major phosphopeptides, one containing phosphoserine and the other phosphothreonine. We have purified the succinylated tryptic phosphopeptides using reverse phase and DEAE high pressure liquid chromatography. The serine-containing peptide, residues 1-4 (Ac-SSKR), is the NH2-terminal peptide. The phosphorylated serine residue may be either serine 1 or serine 2. The threonine-containing peptide, residues 5-16, yielded the sequence AKAKTTKKRPQR. Analysis of the yields and radioactivity of the products from automated Edman degradation showed that threonine 9 is the phosphorylation site.


C-type Lectin Domain Family 4 Member G (CLEC4G) Is a Negative Marker for CD34 in the Evolution of Liver Pathogenesis.
This study aimed to explore the expression of the C-type lectin domain family 4 member G (CLEC4G) gene in the liver sinusoidal endothelial cells (LSECs) during liver pathogenesis, and to evaluate its correlation with CD34 and clinical significance in hepatocellular carcinoma patients. We conducted bioinformatics analysis of the differential expression of CLEC4G in various human organs, carcinomatous and adjacent tissues. Then, mRNA and protein expression levels of CD34 in hepatocellular carcinoma (HCC) samples were detected via real-time quantitative reverse transcription PCR (qRT-PCR) and immunohistochemical (IHC), respectively. ELISA was applied to detect serum levels of CLEC4G in healthy controls, liver fibrosis and HCC patients. The expressions of mRNA and protein levels of CLEC4G were higher in normal liver tissues, moderately expressed in cirrhotic and para-cancerous tissues (P<0.001), and lowest in HCC tissues (P<0.001). We also found high CD34 expression in tumors, which was negatively correlated with CLEC4G at both mRNA and protein levels. Compared to the healthy controls, the CLEC4G levels in liver fibrosis patients and HCC patients gradually became lower (P<0.001). The low expression of CLEC4G is potentially correlated with LSEC capillarization and the appearance of micro-vessels. Such a phenomenon may serve as a reliable diagnostic marker for hepatocellular carcinoma.


Sinonasal cancer: Analysis of oncological failures in 156 consecutive cases.
Sinonasal cancers are rare and associated with a poor prognosis. The purpose of this study was to report our experience and analyze the risk factors for oncologic failures. A retrospective review of 156 consecutive patients treated with curative intent for sinonasal malignancy between 1995 and 2005 at tertiary cancer center was performed. Demographic, clinical, morphological, and pathological parameters were correlated with oncologic outcomes. Complete response was obtained for 134 patients. Sixty-eight patients relapsed, among which 51 had local recurrence. Nine of these 51 patients (17.6%) underwent successful salvage therapy. Five-year local failure and overall survival rates were 50.0% and 61.1%, respectively. Maxillary sinus tumors, intracranial invasion, and N > 0 classification at initial diagnosis were significantly and independently associated with local failure and survival in multivariate analysis. Local control after initial treatment is primordial to optimizing outcomes because of the poor results of salvage therapy.


Persistent polyclonal binucleated B-cell lymphocytosis and MECOM gene amplification.
Persistent Polyclonal Binucleated B-cell Lymphocytosis (PPBL) is characterized by a chronic polyclonal B-cell lymphocytosis with binucleated lymphocytes and a polyclonal increase in serum immunoglobulin-M. Cytogenetic is characterized by the presence of a supernumerary isochromosome +i(3)(q10), premature chromosome condensation and chromosomal instability. Outcome of PPBL patients is mostly benign, but subsequent malignancies could occur. The aim of our study is to provide an update of clinical and cytogenetic characteristics of our large cohort of PPBL patients, to describe subsequent malignancies occurring during the follow-up, and to investigate the role of the long arm of chromosome 3 in PPBL. We analyzed clinical, biological and cytogenetic characteristics (conventional cytogenetic analysis and fluorescent in situ hybridization) of 150 patients diagnosed with PPBL. We performed high-resolution SNP arrays in 10 PPBL patients, comparing CD19(+) versus CD19(-) lymphoid cells. We describe the cytogenetic characteristics in 150 PPBL patients consisting in the presence of supernumerary isochromosome +i(3)(q10) (59%) and chromosomal instability (55%). In CD19(+) B-cells, we observed recurrent copy number aberrations of 143 genes with 129 gains (90%) on 3q and a common minimal amplified genomic region in the MECOM gene. After a median follow-up of 60 months, we observed the occurrence of 12 subsequent malignancies (12%), 6 solid tumors and 6 Non-Hodgkin's Lymphomas, and 6 monoclonal gammopathies of undetermined significance (MGUS), requiring a long-term clinical follow-up. Our clinical and cytogenetic observations lead us to hypothesize that isochromosome 3q, especially MECOM abnormality, could play a key role in PPBL.


Primary familial amyloidosis with vitreous opacities. Report of an autopsy case.
A 41-year-old Japanese male with a new type of primary familial amyloidosis was reported. The patient developed vitreous opacities, and later, disturbances in the gastrointestinal and nervous systems. At autopsy, amyloid was observed in the vitreous and the retinal vessels. There were extensive cerebral infarcts and heavy meningo-vascular amyloid deposition. Although the postmortem study revealed slight peripheral nerve degeneration in the lower extremities secondary to amyloid deposition, there was no clinical evidence of polyneuropathy.


The burden of cancers and their variations across the states of India: the Global Burden of Disease Study 1990-2016.
Previous efforts to report estimates of cancer incidence and mortality in India and its different parts include the National Cancer Registry Programme Reports, Sample Registration System cause of death findings, Cancer Incidence in Five Continents Series, and GLOBOCAN. We present a comprehensive picture of the patterns and time trends of the burden of total cancer and specific cancer types in each state of India estimated as part of the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2016 because such a systematic compilation is not readily available. We used all accessible data from multiple sources, including 42 population-based cancer registries and the nationwide Sample Registration System of India, to estimate the incidence of 28 types of cancer in every state of India from 1990 to 2016 and the deaths and disability-adjusted life-years (DALYs) caused by them, as part of GBD 2016. We present incidence, DALYs, and death rates for all cancers together, and the trends of all types of cancers, highlighting the heterogeneity in the burden of specific types of cancers across the states of India. We also present the contribution of major risk factors to cancer DALYs in India. 8·3% (95% uncertainty interval [UI] 7·9-8·6) of the total deaths and 5·0% (4·6-5·5) of the total DALYs in India in 2016 were due to cancer, which was double the contribution of cancer in 1990. However, the age-standardised incidence rate of cancer did not change substantially during this period. The age-standardised cancer DALY rate had a 2·6 times variation across the states of India in 2016. The ten cancers responsible for the highest proportion of cancer DALYs in India in 2016 were stomach (9·0% of the total cancer DALYs), breast (8·2%), lung (7·5%), lip and oral cavity (7·2%), pharynx other than nasopharynx (6·8%), colon and rectum (5·8%), leukaemia (5·2%), cervical (5·2%), oesophageal (4·3%), and brain and nervous system (3·5%) cancer. Among these cancers, the age-standardised incidence rate of breast cancer increased significantly by 40·7% (95% UI 7·0-85·6) from 1990 to 2016, whereas it decreased for stomach (39·7%; 34·3-44·0), lip and oral cavity (6·4%; 0·4-18·6), cervical (39·7%; 26·5-57·3), and oesophageal cancer (31·2%; 27·9-34·9), and leukaemia (16·1%; 4·3-24·2). We found substantial inter-state heterogeneity in the age-standardised incidence rate of the different types of cancers in 2016, with a 3·3 times to 11·6 times variation for the four most frequent cancers (lip and oral, breast, lung, and stomach). Tobacco use was the leading risk factor for cancers in India to which the highest proportion (10·9%) of cancer DALYs could be attributed in 2016. The substantial heterogeneity in the state-level incidence rate and health loss trends of the different types of cancer in India over this 26-year period should be taken into account to strengthen infrastructure and human resources for cancer prevention and control at both the national and state levels. These efforts should focus on the ten cancers contributing the highest DALYs in India, including cancers of the stomach, lung, pharynx other than nasopharynx, colon and rectum, leukaemia, oesophageal, and brain and nervous system, in addition to breast, lip and oral cavity, and cervical cancer, which are currently the focus of screening and early detection programmes. Bill & Melinda Gates Foundation; and Indian Council of Medical Research, Department of Health Research, Ministry of Health and Family Welfare, Government of India.


Measurement of functional residual capacity of the lung by partial CO2 rebreathing method during acute lung injury in animals.
Several techniques for measuring the functional residual capacity (FRC) of the lungs in mechanically ventilated patients have been proposed, each of which is based on either nitrogen wash-out or dilution of tracer gases. These methods are expensive, difficult, time-consuming, impractical, or require an intolerably large change in the fraction of inspired oxygen. We propose a CO(2) wash-in method that allows automatic and continual FRC measurement in mechanically ventilated patients. We measured FRC with a CO(2) partial rebreathing technique, first in a mechanical lung analog, and then in mechanically ventilated animals, before, during, and subsequent to an acute lung injury induced with oleic acid. We compared FRC measurements from partial CO(2) rebreathing to measurements from a nitrogen wash-out reference method. Using an approved animal protocol, general anesthesia was induced and maintained with propofol in 6 swine (38.8-50.8 kg). A partial CO(2) rebreathing monitor was placed in the breathing circuit between the endotracheal tube and the Y-piece. The partial CO(2) rebreathing signal obtained from the monitor was used to calculate FRC. FRC was also measured with a nitrogen wash-out measurement technique. In the animal studies we collected data from healthy lungs, and then subsequent to a lung injury that simulated the conditions of acute lung injury/acute respiratory distress syndrome. The injury was created by intravenously infusing 0.09 mL/kg of oleic acid over a 15-min period. At each stage of the experiment, the positive end-expiratory pressure (PEEP) was set to 0, 5, 10, or 15 cm H(2)O. At each PEEP level we compared the average of 3 CO(2) rebreathing FRC measurements to the average of 3 nitrogen wash-out reference measurements. We also tested the FRC measurement system with a mechanical test lung in which the true FRC could be directly measured. The squared correlation for the linear regression between CO(2) rebreathing and nitrogen wash-out measurements in the animals was r(2) = 0.89 (n = 50). The average error of the CO(2) wash-out system was -87 mL and the limits of agreement were+/- 263 mL. In the mechanical test lung, the average error of the FRC measured via the CO(2) wash-in system was 37 mL, and the limits of agreement were +/- 103 mL, which was equivalent to 1.7% of the true FRC. The squared correlation was r(2) = 0.96. These results indicate that FRC measurement via CO(2) rebreathing can reliably detect an FRC decrease during lung injury and can reflect the response of the FRC to treatment with PEEP.


Specific conversion of s4 U to U in Escherichia coli tRNA by iodate oxidation.
The minor nucleoside 4-thiouridine in Escherichia coli tRNA is transformed selectively to uridine by iodate oxidation at acidic pH. The four major nucleotides were found to be inert under these conditions. The iodate oxidation appears to be more specific than the previous conversion methods reported, and has the advantage that it does not affect the chargeability of most tRNA.


Conjugated linoleic acid promotes human adipocyte insulin resistance through NFkappaB-dependent cytokine production.
We previously demonstrated that trans-10, cis-12 conjugated linoleic acid (CLA) reduced the triglyceride content of human adipocytes by activating mitogen-activated protein kinase kinase/extracellular signal-related kinase (MEK/ERK) signaling via interleukins (IL) 6 and 8. However, the upstream mechanism is unknown. Here we show that CLA increased (>or=6 h) the secretion of IL-6 and IL-8 in cultures containing both differentiated adipocytes and stromal vascular (SV) cells, non-differentiated SV cells, and adipose tissue explants. CLA isomer-specific induction of IL-6 and tumor necrosis factor-alpha was associated with the activation of nuclear factor kappaB (NFkappaB) as evidenced by 1) phosphorylation of IkappaBalpha, IkappaBalpha kinase, and NFkappaB p65, 2) IkappaBalpha degradation, and 3) nuclear translocation of NFkappaB. Pretreatment with selective NFkappaB inhibitors and the MEK/ERK inhibitor U0126 blocked CLA-mediated IL-6 gene expression. Trans-10, cis-12 CLA suppression of insulin-stimulated glucose uptake at 24 h was associated with decreased total and plasma membrane glucose transporter 4 proteins. Inhibition of NFkappaB activation or depletion of NFkappaB by RNA interference using small interfering NFkappaB p65 attenuated CLA suppression of glucose transporter 4 and peroxisome proliferator-activated receptor gamma proteins and glucose uptake. Collectively, these data demonstrate for the first time that trans-10, cis-12 CLA promotes NFkappaB activation and subsequent induction of IL-6, which are at least in part responsible for trans-10, cis-12 CLA-mediated suppression of peroxisome proliferator-activated receptor gamma target gene expression and insulin sensitivity in mature human adipocytes.


Effect of Nigella sativa oil on experimental toxoplasmosis.
Toxoplasmosis is a parasitic infection caused by Toxoplasma gondii protozoon. It is most commonly treated by pyrimethamine (PYR); however, this was intolerable by many patients. The aim of this study was to assess therapeutic effects of Nigella sativa oil (NSO) alone and combined with pyrimethamine (PYR) compared to a previous combination of clindamycin (CLN) and (PYR). One hundred Albino mice were used in the current study and were equally divided into five groups: normal (I), infected untreated control (II); infected, treated with NSO-only (III); infected, treated with NSO + PYR (IV); and infected, treated with CLN + PYR (V). The virulent RH Toxoplasma strain was used in infection survival rates estimation, impression smears from liver and spleen, and histopathological and ultrastructural studies were done. Liver malondialdehyde (MDA) level and total antioxidant capacity (TAC) were determined. Interferon-γ and specific IgM were also measured in sera by ELISA. Results showed that NSO alone has no direct anti-Toxoplasma effect, whereas its combination with PYR produced potent effect that is comparable to CLN + PYR. It significantly increased the survival rate and decreased the parasite density and pathological insult in both liver and spleen. Also, significant increase in interferon-γ level denotes stimulation of cellular immunity. NSO + PYR combination markedly improved the antioxidant capacity of Toxoplasma infected mice compared to the infected untreated ones and to CLN/PYR. In conclusion, although NSO, if administered alone, has significant immunostimulant and antioxidant properties, it failed to decrease tachyzoite counts. Combination of NSO and PYR had synergistic effect in treatment of toxoplasmosis.


Synthesis and an X-ray structure of soluble phenylacetylene macrocycles with two opposing bipyridine donor sites
The synthesis of the shape-persistent macrocycles 10a and 10b with two bipyridine units in opposing sides by Hagihara/Sonogashira cross-coupling chemistry of suitably functionalized building blocks is reported. X-ray analysis of single crystals of 10b shows a layered structure with channels filled with solvent molecules and parts of the flexible chains. with which the cycle is decorated for solubility reasons.


Impact of nitrite exposure on oxidative stress and antioxidative-related genes responses in the gills of Procambarus clarkii.
Nitrite is the major environmental pollutant in the freshwater aquaculture environment, which has a negative impact on aquatic species growth. Currently, we know that the main way nitrite enters crustaceans is through their gills. In this study, a total of 96 h acute nitrite stress (60 mg/L) experiments were conducted, and the impact of the serum biochemical parameters, gill oxidase activity and oxidative-related gene expression of red swamp crayfish were evaluated. After exposure to nitrite for 0, 6, 12, 24, 48, and 96 h, hemolymph and gills samples were taken at each time point. In the serum, acute nitrite stress significantly increased glutamic-oxaloacetic transaminase (GOT) and alanine aminotransferase (ALT) activities after 6 h of exposure, decreased total protein (TP) and albumin (ALB) levels after 24 h and 48 h of exposure, respectively. In the gills, the activities of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were enhanced to the maximum level at 12 h, 24 h and 24 h, respectively. The contents of malondialdehyde (MDA) and lipid peroxide (LPO) were increased significantly after 12 h and 24 h exposure, respectively. In addition, the expression levels of antioxidative-related genes, including hsp70, fer and mt, were significantly upregulated in the gills after 6 h of exposure. The results indicated that acute nitrite stress changed the serum physiological status, induced oxidative stress and caused damage to gill cells in P. clarkii.


Effect of substituting saturated with monounsaturated fatty acids on serum visfatin levels and insulin resistance in overweight women: a randomized cross-over clinical trial.
This study aimed to determine the effects of a monounsaturated fatty acids (MUFA)-rich diet on serum visfatin, interleukin-6 and insulin levels among overweight women. In this randomized cross-over clinical trial, 17 premenopausal overweight women were included. Participants were randomly assigned to consume either a hypocaloric, saturated fatty acids (SFA)-rich diet (16% SFA and 8% MUFA) or a hypocaloric, MUFA-rich diet (16% MUFA and 8% SFA) for 12 weeks crossing over after a 2-week washout period. Percentages of energy from other macronutrients were similar between the two diets. Biochemical and anthropometric assessments were done at the first and at the end of each period. Statistical analyses were done using paired t-test. In all statistical analysis, p < 0.05 was considered as significant. The participant's mean body mass index was 27.6 kg/m². Mean percentages of MUFA intake were 13% during MUFA-rich diet and 7% during SFA-rich diet. The corresponding values for SFA intake were 8.5% and 14%, respectively. We failed to find any significant differences between two intervention diets in terms of their effect on the serum levels of IL-6, visfatin and insulin. However, serum visfatin and IL-6 levels increased during the SFA-rich diet (0.4 ± 0.4 ng/ml and 0.19 ± 0.3 pg/ml, respectively) and decreased during the MUFA-rich diet (-0.7 ± 0.5 ng/ml and -0.17 ± 0.3 pg/ml, respectively). In spite of a slight reduction in both periods, changes in serum insulin levels did not reach significant levels comparing the two periods. Our findings did not support any significant effect of a MUFA-rich intake on serum IL-6 and insulin levels as compared with a SFA-rich diet; however, it has the potential to favourably affect serum visfatin levels.


Implications of quantal motor action in biological systems.
We demonstrate in this paper that quantal behavior is a central feature of biological motile and contractile systems. Step-like behavior has been demonstrated in the interaction between single molecules and filaments both in the kinesin-microtubule system and in the myosin-actin filament system. We show here that the step-like molecular features appear also in the single intact sarcomere. We studied single sarcomeres of single bumble-bee myofibrils, both in the unactivated and activated states. Myofibril-length changes induced by a motor-imposed ramp were accompanied by corresponding sarcomere-length changes. However, the sarcomere-length changes were stepwise. Computer analysis of the stepwise shortening patterns revealed a step-size distribution containing multiple peaks. In the activated state, the peaks were separated by 2.7 nm per half-sarcomere which is the linear actin-subunit spacing. Thus, translation steps are an integer multiple of the actin-subunit spacing. This result parallels the one observed in the kinesin-tubulin spacing, where step size is a multiple of the tubulin-subunit spacing. In the muscle system, however, the steps are preserved on a macroscopic scale, implying high synchrony. The quantal steps are easily explained by a model in which the actin filament propels itself over stationary cross-bridges: if actin binds to the cross-bridges between steps, then the observed quantal result is inevitable. As probes of contractile phenomena approach the molecular level, the discrete unitary events underlying contraction begin to emerge. Thus, step-like behavior is observed as the single kinesin molecule translates along the microtubule, as the single myosin molecule translates over the actin filament, and as the single isolated titin molecule is stretched.


Does hair cell differentiation predate the vertebrate appearance?
It is generally accepted that the three main chordate groups (tunicates, cephalochordates and vertebrates) originated from a common ancestor having the basic features of the chordate body plan, i.e. a neural tube and a notochord flanked by striated musculature. There is now increasing evidence that tunicates, rather than cephalochordates, are the vertebrate sister-group. Correlated with this, tunicates have sensory structures similar to those derived from placodes or neural crest in vertebrates. In this context, we discuss here whether the precursors of vertebrate hair cells, which are placodal in origin, were present in ancestral chordates. The ascidian tunicates possess a coronal organ, consisting of a row of mechanosensory cells that runs around the base of the oral siphon. Its function is to monitor the incoming water flow. The cells are secondary sensory cells, i.e. they lack axons and synapse with neurons whose somata lie in the cerebral ganglion. They are accompanied by supporting cells and, as in vertebrates, have varying morphologies in the species so far examined: in one order (Enterogona), they are multiciliate; in the other (Pleurogona), they may possess an apical apparatus, consisting of one or two cilia accompanied by stereovilli, that are graded in length. Coronal cells thus resemble vertebrate hair cells closely in their morphology, embryonic origin and arrangement, which suggests they originated early in ancestral chordates. We are continuing our study of the coronal organ in other ascidian species, and report new data here on Botrylloides leachi, which conforms with the pattern of Pleurogona and, in particular, with previously published results on other botryllid ascidians.


Selective effects of thyroid hormonal deprivation on growth and development of olfactory receptor sheet during the early postnatal period: a morphometric and cell count study in the rat.
Light microscopic numerical and morphometric studies were conducted on the olfactory epithelium of postnatal normal and hypothyroid rats. The normal rat olfactory epithelium undergoes marked growth and development during the suckling period (days 1-25): thickness, 50%; area, x 8, total number of olfactory neurons, basal and supporting cells, x 10, x 11 and x 8, respectively. The effects of thyroid hormonal deprivation on these proliferative postnatal growth changes were studied by adding PTU (n-propylthiouracil, a reversible antithyroid goitrogen) to the litter's drinking water from birth to weaning (day 25). The general architecture of naso-olfactory cavities as well as the histology and thickness of the olfactory epithelium were unaffected in the hypothyroid pups. However, the surface area of the olfactory receptor sheet was reduced by 40%, the reduction occurring throughout the cavity, though not uniformly. The total number of olfactory neurons, supporting and basal cells were reduced by 33, 45 and 47%, respectively. These results indicate that the postnatal vertical accretion of olfactory neurons occurring across the epithelial thickness is unaffected in the hypothyroid pups, while the horizontal proliferation of neurons accompanying the expansion of the sheet's surface area is markedly reduced. The results suggest differential effects of thyroid hormones on these modes of proliferative growth and imply further that in addition to possible direct effects, the influence of thyroid hormones on developmental growth of the olfactory epithelial sheets may be secondary to effects on the underlying submucosal connective tissue.


Enantioselective Conversion of Achiral Cyclohexadienones to Chiral Cyclohexenones by Desymmetrization.
The enantioselective reduction of prochiral 4,4-disubstituted 2,5-cyclohexadienones to chiral 2-cyclohexenones has been accomplished by the use of a carefully selected chiral bisphosphine-CuI complex and diisobutylaluminum hydride-hexamethylphosphoric triamide complex. This reagent has provided access to a key bicyclic intermediate for the total synthesis of the natural enantiomer of the pentacyclic sesterterpene retigeranic acid that involves spatial discrimination between CH3 and CH2CH2R substituents, an operation that has been elusive previously. In addition, a second method for desymmetrization is described using catalytic enantioselective [4 + 2]-cycloaddition of cyclopentadiene to prochiral 4,4-disubstituted 2,5-cyclohexadienones.


Cholinesterases within neurofibrillary tangles related to age and Alzheimer's disease.
Enzyme histochemistry showed that a substantial number of cortical neurofibrillary tangles contain acetylcholinesterase and butyrylcholinesterase activities in patients with Alzheimer's disease and in nondemented aged individuals. In some areas of the brain, tangles containing each of these two enzymes were differentially distributed. In other areas, there was a very close overlap between the two types of cholinesterase.


Rapidly progressive organizing pneumonia associated with COVID-19.
We report a case of clinically diagnosed secondary organizing pneumonia (SOP) associated with coronavirus disease 2019 (COVID-19). A 70-year-old woman who had been diagnosed with COVID-19 was admitted to Hokkaido University Hospital. Although her fever, cough, dyspnea, and serum C-reactive protein levels improved, she developed rapidly progressive respiratory failure and computed tomography revealed the development of bilateral lung consolidation. Her dyspnea was relieved, and her oxygenation levels and radiological findings improved after commencing corticosteroid treatment. Blood biomarkers for interstitial lung disease, Krebs von den Lungen-6 (KL-6) and surfactant protein D (SP-D), showed different responses during the clinical course of her disease. Evaluation of serial changes in levels of KL-6 and SP-D may help diagnose and monitor COVID-19-associated organizing pneumonia (OP). Clinicians should be aware that SOP can develop in response to COVID-19 and that these patients may benefit from the use of steroids.


[Proximal femoral fractures in the German external quality assurance module 17/1: reasons for delay in surgery].
For the quality indicator "preoperative stay" a part of the external quality assurance for proximal femoral fractures (module 17/1), a tolerance range for surgery within 48 h after admission of ≤ 15 % is given. Over a period of 5 years all cases were analyzed with respect to reasons for delayed surgery of more than 48 h after admission. A total of 165 patients (16%) out of 1,036 documented cases had surgery later than 48 h after admission. Reasons were pathological bleeding, preoperative poor general condition, lack of informed consent, intake of metformin and lacking initial radiological detection of fractures. Due to a lack of software-related specifications in ten patients a wrong preoperative length of stay was generated. The significance of the quality indicator "preoperative stay" without division into whether this was administrative or patient-related must be considered critically. For fall-related fractures in hospital the time of the accident or diagnosis should be considered.


Yes-associated protein regulates cortical actin architecture and dynamics through intracellular translocation of Rho GTPase-activating protein 18.
Yes-associated protein (YAP) is a transcriptional co-activator that controls the transcription of target genes and modulates the structures of various cytoskeletal architecture as mechanical responses. Although it has been known that YAP regulates actin-regulatory proteins, the detailed molecular mechanism of how they control and coordinate intracellular actin architecture remains elusive. Herein, we aimed to examine the structure and dynamics of intracellular actin architecture from molecular to cellular scales in normal and YAP-knockout (YAP-KO) cells utilizing high-speed atomic force microscopy (HS-AFM) for live-cell imaging and other microscope-based mechanical manipulation and measurement techniques. YAP-KO Madin-Darby canine kidney cells had a higher density and turnover of actin filaments in the cell cortex and a higher elastic modulus. Laser aberration assay demonstrated that YAP-KO cells were more resistant to damage than normal cells. We also found that Rho GTPase-activating protein 18 (ARHGAP18), a downstream factor of YAP, translocated from the cortex to the edge of sparsely cultured YAP-KO cells. It resulted in high RhoA activity and promotion of actin polymerization in the cell cortex and their reductions at the edge. HS-AFM imaging of live cell edge and a cell-migration assay demonstrated lower membrane dynamics and motility of YAP-KO cells than those of normal cells, suggesting lower actin dynamics at the edge. Together, these results demonstrate that a YAP-dependent pathway changes the intracellular distribution of RhoGAP and modulates actin dynamics in different parts of the cell, providing a mechanistic insight into how a mechano-sensitive transcription cofactor regulates multiple intracellular actin architecture and coordinates mechano-responses.


Interplay between DNA and RNA Modifications: A Constantly Evolving Process.
The epigenome refers to the entirety of DNA methylations, histone modifications, nucleosome occupancy, and coding and non-coding RNAs (and their modifications) in different cell types [...].


Prediction of memory rehabilitation outcomes in traumatic brain injury by using functional magnetic resonance imaging.
To evaluate the ability of functional magnetic resonance imaging (fMRI) measures collected from people with traumatic brain injury (TBI) to provide predictive value for rehabilitation outcomes over and above standard predictors. Prospective study. Academic medical center. Persons (N=54) with TBI greater than 1 year postinjury. A novel 12-session group rehabilitation program focusing on internal strategies to improve memory. The Hopkins Verbal Learning Test-Revised (HVLT-R) delayed recall score. fMRI measures were collected while participants performed a strategically directed word memorization task. Prediction models were multiple linear regressions with the following primary predictors of outcome: age, education, injury severity, preintervention HVLT-R, and task-related fMRI activation of the left dorsolateral and left ventrolateral prefrontal cortex (VLPFC). Baseline HVLT-R was a significant predictor of outcome (P=.007), as was injury severity (for severe vs mild, P=.049). We also found a significant quadratic (inverted-U) effect of fMRI in the VLPFC (P=.007). This study supports previous evidence that left prefrontal activity is related to strategic verbal learning, and the magnitude of this activation predicted success in response to cognitive memory rehabilitation strategies. Extreme under- or overactivation of VLPFC was associated with less successful learning after rehabilitation. Further study is necessary to clarify this relationship and to expand and optimize the possible uses of functional imaging to guide rehabilitation therapies.


Increased forefoot loading is associated with an increased plantar flexion moment.
The aim of this study was to identify the cascade of effects leading from alterations in force generation around the ankle joint to increased plantar pressures under the forefoot. Gait analysis including plantar pressure measurement was performed at an individually preferred and a standardized, imposed gait velocity in diabetic subjects with polyneuropathy (n=94), without polyneuropathy (n=39) and healthy elderly (n=19). The plantar flexion moment at 40% of the stance phase was negatively correlated with the displacement rate of center of pressure (r=-.749, p<.001 at the imposed, and r=-.693, p<.001 at the preferred gait velocity). Displacement rate of center of pressure was strongly correlated with forefoot loading (r=-.837, p<.001 at the imposed, and r=-.731, p<.001 at the preferred gait velocity). People with a relatively high plantar flexion moment at 40% of the stance phase, have a faster forward transfer of center of pressure and consequently higher loading of the forefoot. This indicates that interventions aimed at increasing the control of the roll-off of the foot may contribute to a better plantar pressure distribution.


Bone conduction masking for brainstem auditory-evoked potentials (BAEP) in pediatric audiological evaluations. Validation of the test.
A brainstem auditory-evoked potential (BAEP) protocol for testing pediatric patients at risk for conductive hearing impairment was evaluated. The protocol used was: air-conducted click stimuli masked by bone-conducted wide-band noise. The specificity and sensitivity values for the test were determined by means of a blind cross-sectional trial including an active group of patients with an aural malformation and an age-matched control group with a sensorineural impairment. The bone-conducted masking of air-conducted BAEP showed high specificity and sensitivity and was easily administered despite pediatric difficulty. It was useful in differentiating sensorineural from conductive impairment and provided a rough estimate of the cochlear reserve in presumptive conductive hearing loss as great as 60 dB hearing loss. It is concluded that the bone-conducted masking procedure appears to be a great help in the binary decision whether middle ear surgery should be performed in patients at risk for conductive hearing loss, specially children with aural malformations.


Green islands - predation not nutrition.
The existence of green islands around wood-ant nests in otherwise damaged birch forests has been explained by two alternative hypotheses: (1) predation by ants protects the trees against defoliators, and (2) the ants ameliorate tree vigor by concentrating soil nutrients. The size of green islands and the nitrogen content of the soil and foliage do not support the nutrition hypothesis. These data and general knowledge about the foraging strategy of wood ants are consistent with the predation hypothesis.


The clinical utility of using Catrimox-14-treated whole blood in detecting hepatitis C virus RNA.
Measuring hepatitis C virus (HCV) RNA in serum or plasma may underestimate the true HCV burden. Extracting viral RNA from whole blood (WB) with a cationic surfactant (Catrimox-14) has resulted in HCV RNA concentrations up to 1000-fold higher than from serum or plasma in some studies, but not others. We compared the Catrimox-14 WB assay with a standard serum assay. Seventy-two chronic HCV patients received 48 weeks of standard or pegylated interferon alpha-2b and ribavirin therapy. Catrimox-14-treated WB and corresponding serum samples were obtained at baseline and weeks 12, 24, 48 and 72. HCV RNA concentrations from WB and serum were quantified by a previously validated RT-PCR assay. Overall mean (+/- SD) baseline serum log10 HCV RNA concentration was 6.5 ((+/- 0.58) copies/ml. Out of 72 patients, 33 had no detectable virus at 72 weeks. Neither assay detected virus in these patients at 12 weeks and neither WB nor serum assays detected virus at end-of-treatment in the 10 patients who subsequently relapsed at 72 weeks. HCV RNA concentrations from WB and serum assays were linearly correlated (R2 = 0.73; P < 0.001), although mean serum HCV RNA concentrations were 0.5 log10, copies/ml higher in serum than in WB [6.0 (+/- 0.82) vs 5.5 ((+/- 0.84), respectively, P = 0.12]. Catrimox-14-treated WB assays detect changes in HCV RNA, but do not offer clinical advantage over a conventional serum RT-PCR based assay.


Learning the deictic meaning of third person pronouns.
Forty-two children (2;8-5;7) viewed video-taped "scenes' in which a SPEAKER addressed sentences containing a personal pronoun which referred to him/herself, the ADDRESSEE or a male or female OTHER. The child's task was to identify the referent of the pronoun. No other cues to the identity of the referent were given. Errors showed that initially gender was the variable used to identify referents and that person was used only later. Several of the oldest children still used only gender in some types of utterance situations. This was because the deictic roles OTHER and, to some extent, ADDRESSEE, were not fully understood until relatively late. Possible explanations concerned the use of Baby Talk to children and information processing necessary for referent identification.


Computerized artifact detection for ventilatory inductance plethysmographic apnea monitors.
Ventilatory inductive plethysmography allows noninvasive monitoring of patient ventilation. Patient movements unrelated to breathing introduce severe errors in ventilator inductive plethysmographic measurements and restrict its usefulness. The purpose of this research was to develop and test a microprocessor-based real-time digital signal processor that uses an adaptive filter to detect patient movements unrelated to breathing. The adaptive filter processor was tested for retrospective identification of artifacts in 20 male volunteers who performed the following specific movements between epochs of quiet, supine breathing: raising arms and legs (slowly, quickly, once, and several times), sitting up, breathing deeply and rapidly, and rolling from a supine to a lateral decubitus position. Flow was simultaneously measured directly with a pneumotachography attached to a mouthpiece. A multilinear regression was used to continuously calculate the calibration constants that relate the pneumotachographic and ventilatory inductive plethysmographic signals. Ventilatory inductive plethysmographic data were then processed, and results scored. There were a total of 166 movements. The calibration coefficients changed dramatically in 146 (88%) of the 166 movements. These movements would have significant errors on ventilatory inductive plethysmographic flow calculation. The changes lasted for the duration of the movements and returned to baseline within two to three breaths. The changes in the coefficients were five or more times larger than the variability around baseline during quiet, supine breathing. All of the total body movements and changes in breathing patterns were detected accurately. The filter detected 46 of 53 upper body movements, 34 of 36 lower body movements, 38 of 38 total body movements, and 19 of 19 breathing pattern changes where the calibration changed.(ABSTRACT TRUNCATED AT 250 WORDS)


Expression of 2A peptide mediated tri-fluorescent protein genes were regulated by epigenetics in transgenic sheep.
A number of gene therapy applications and basic research would benefit from vectors expressing multiple genes. In this study, we constructed 2A peptide based tricistronic lentiviral vector and generated transgenic lambs by injecting lentivirus carrying the tricistronic vector into perivitelline space of zygotes. Of 7 lambs born, 2 lambs (#6 and #7) carried the transgene. However, no fluorescent proteins were identified in transgenic sheep. To investigate why the transgene was silenced in transgenic sheep, we analyzed the methylation status of transgene. The methylation level of CMV promoter was 76.25% in #6, and 64.7% in #7. In the coding region of three fluorescent protein genes, methylation levels were extremely high, with the average level of 98.3% in #6 and 98.4% in #7 respectively. Furthermore, the ratio of GFP(+) cells were increased significantly when the fibroblasts derived from the transgenic sheep were treated with 5-azaC and/ or TSA. Our results showed that 2A peptide based tricistronic construct was subjected to hypermethylation in transgenic sheep. Moreover, the silencing could be relieved by treating with methytransferase inhibitor and/or deacetylase inhibitor.


Synergist effect of sucrose fatty acid esters on nisin inhibition of gram-positive bacteria.
Nisin in combination with the sucrose fatty acid esters, sucrose palmitate (P-1570 and P-1670) or sucrose stearate (S-1570 and S-1670) was tested against a range of Gram-negative and Gram-positive bacteria. Initial liquid culture investigation showed that the sugar ester P-1670 resulted in a synergist enhancement of the bacteriostatic activity of nisin against Gram-positive bacteria and not Gram-negative bacteria. Some enhancement of the bactericidal activity of nisin against Listeria monocytogenes was also observed. This increased nisin inhibitory effect was confirmed on solid media using plates with gradients of pH and NaCl. Synergism was observed with all four sucrose fatty acid esters, which enhanced the antimicrobial activity of nisin against several strains of L. monocytogenes, Bacillus cereus (both cells and spores), Lactobacillus plantarum and Staphylococcus aureus. The combination of nisin and the sucrose fatty acid esters showed no inhibition of Gram-negative bacteria (Salmonella enteritidis, Salm. typhimurium and Pseudomonas aeruginosa).


Radiochemical characterization of produced water from two production offshore oilfields in Ghana.
Produced water from two Ghanaian offshore production oilfields has been characterized using alpha spectrometry after radiochemical separation, non-destructive gamma spectrometry and ICP-MS and other complimentary analytical tools. The measured concentrations of main NORM components were in the range of 6.2-22.3 Bq.L(-1), 6.4-35.5 Bq.L(-1), and 0.7-7.0 Bq.L(-1) for (226)Ra, (228)Ra and (224)Ra respectively. A good correlation between several physico-chemical parameters and radium isotopes was observed in each production oilfield. The radium concentrations obtained in this study for produced water from the two oilfields of Ghana are of radiological importance and hence there may be the need to put in place measures for future contamination concerns due to their bioavailability in the media and bioaccumulation characteristics. The results will assist in critical decision making for future set up of appropriate national guidelines for the management of NORM waste from the emerging oil and gas industry in Ghana.


The efficacy of trivalent cyclic hexapeptides to induce lipid clustering in PG/PE membranes correlates with their antimicrobial activity.
Various models have been proposed for the sequence of events occurring after binding of specific antimicrobial peptides to lipid membranes. The lipid clustering model arose by the finding that antimicrobial peptides can induce a segregation of certain negatively charged lipids in lipid model membranes. Anionic lipid segregation by cationic peptides is initially an effect of charge interaction where the ratio of peptide and lipid charges is thought to be the decisive parameter in the peptide induced lipid demixing. However, the sequence of events following this initial lipid clustering is more complex and can lead to deactivation of membrane proteins involved in cell division or perturbation of lipid reorganization essential for cell division. In this study we used DSC and ITC techniques to investigate the effect of binding different cyclic hexapeptides with varying antimicrobial efficacy, to phosphatidylglycerol (PG)/phosphatidylethanolamine (PE) lipid membranes and their ability to induce lipid segregation in these mixtures. We found that these cyclic hexapeptides consisting of three charged and three aromatic amino acids showed indeed different abilities to induce lipid demixing depending on their amino acid composition and their sequence. The results clearly showed that the cationic amino acids are essential for electrostatic binding but that the three hydrophobic amino acids in the peptides and their position in the sequence also contribute to binding affinity and to the extent of induction of lipid clustering. The efficacy of these different hexapeptides to induce PG clusters in PG/PE membranes was found to be correlated with their antimicrobial activity.


The effect of hormone therapy on quality of life and breast cancer risk after risk-reducing salpingo-oophorectomy: a systematic review.
It is unclear if the use of hormone therapy (HT) in carriers of BRCA mutations improves the quality of life (QOL) without increasing the risk of breast cancer following a risk-reducing salpingo-oophorectomy (RRSO). Our objective was to assess the effect of HT on QOL and breast cancer risk, after RRSO. We searched MEDLINE, EMBASE, CINHAL, and others, from inception to July 22, 2016, to identify relevant studies. Two reviewers independently screened identified records for controlled trials and observational studies that addressed the effect of HT on QOL and breast cancer risk in women with BRCA mutations, post RRSO. Two reviewers independently extracted data on populations, interventions, comparators, outcomes, and methodological quality. Studies addressing the same outcome were synthesized using written evidence summaries or tables. Of the 1,059 records identified, 13 met our inclusion criteria. All studies were observational. Six studies assessed the effect on QOL. Of these, 3 showed improvement in QOL with HT use. The risk of breast cancer was evaluated in 4 studies. The mean duration of follow-up was 2.6 years (range 0.1-19.1). The risk of breast cancer did not change with HT use in all 4 studies. Cumulative evidence from our review suggests that short-term HT use following RRSO improves QOL. The effect on breast cancer risk is still unclear. There are too few long-term studies to draw any strong conclusions. The need for future well-designed RCTs for more established evidence is imperative.


Fermented infant formulas without live bacteria: a systematic review.
Fermented formulas, i.e., those fermented with lactic acid-producing bacteria during the production process and not containing significant amounts of viable bacteria in the final product, are widely available in many countries. Our aim was to systematically review published evidence related to the safety and health effects of the administration of fermented infant formulas compared with standard infant formulas. The Cochrane Library, MEDLINE, and EMBASE databases and major pediatric conference proceedings were searched. Five randomized controlled trials (RCTs) involving 1326 infants met the inclusion criteria. Compared with standard formula, the use of fermented formula resulted in a similar weight gain and length gain during the study period. Data from one RCT, albeit large, suggest the effectiveness of fermented formula in preventing and treating acute diarrhea. Fermented formula has the potential to reduce some, albeit not well-defined, digestive symptoms. Current evidence does not support the use of fermented formula for preventing cow's milk allergy. Limited available evidence suggests that the use of fermented infant formula, compared with the use of standard infant formula, does not offer clear additional benefits, although some benefit on gastrointestinal symptoms cannot be excluded. What is known • Fermented formulas, i.e., those fermented with lactic acid-producing bacteria during the production process and not containing significant amounts of viable bacteria in the final product, are widely available in many countries. What is new • Limited evidence available suggests that the use of fermented infant formula, compared with the use of standard infant formula, does not offer clear additional benefits, although some benefit on gastrointestinal symptoms cannot be excluded. At the same time, no negative health effects have been documented.


Factors Associated with Greater Pain Intensity.
The intensity of pain reported for a given nociception is highly variable. Variation in pain intensity is best accounted for by stress, distress, and ineffective coping strategies. Among orthopedic surgery patients, greater intake of opioids is associated with greater pain intensity and decreased satisfaction with pain control, no matter the pathophysiology or nociception. The single most effective pain reliever is self-efficacy (the sense that one can manage and that everything will be okay).


Effects of Reading Interventions Implemented for Upper Elementary Struggling Readers: A Look at Recent Research.
In this study, we conducted a review of reading intervention research (1988-2019) for upper elementary struggling readers and examined intervention area (e.g., foundational, comprehension, or multicomponent) and intensity (e.g., hours of intervention, group size, and individualization) as possible moderators of effects. We located 33 studies containing 49 treatment-comparison contrasts, found small effects for foundational reading skills (g = 0.22) and comprehension (g = 0.21), and decreased effects when considering standardized measures only. For intervention area, only multicomponent interventions predicted significant effects for both comprehension and foundational outcomes. For intensity, we did not find systematic evidence that longer or individualized interventions were associated with larger effects. However, interventions implemented in very small groups predicted larger comprehension outcomes. Overall, more research examining the quality of school provided reading instruction and how the severity of reading difficulties may impact effects of more intensive interventions is needed.


Expression of the mouse homologue for the human GCDFP-15/PIP gene during pre- and early post-natal development.
The function of the mouse submaxillary gland/prolactin inducible protein (mSMGP/mPIP), the homologue of the human gross cystic disease fluid protein 15 (GCDFP-15)/prolactin inducible protein (hPIP) remains unknown. The human gene, normally expressed in apocrine glands of healthy individuals, is aberrantly expressed in human breast cancers where it is regulated by hormones including androgens, and in prostate cancers. We have previously reported that in the adult mouse and rat, gene expression is tissue-specific for the salivary and lacrimal glands, and is hormonally regulated. In this study, we examine the endogenous pattern of mouse SMGP/PIP (mSMGP/mPIP) gene expression in mid- and late-embryonic, and in early postnatal development. Gene expression was analyzed by RT-PCR followed by Southern blot analysis, and by in situ hybridization. Gene expression was detected in the submandibular gland as early as embryonic day 14 (E14), a period that coincides with the initiation of submandibular gland development in the embryo, suggesting that mSMGP/mPIP may have a functional role in the developing gland. Nearing the end of gestation, E18, mSMGP/mPIP transcripts were localized in the proacinar cells of the gland, and gene expression continued to be maintained following birth. In addition, during early postnatal development, mSMGP/mPIP gene expression was detected in the other two major salivary glands, the sublingual and parotid, as well as in the lacrimal gland and in reproductive tissues. In the prostate, gene expression was turned off by 10 weeks of age. The spatial and temporal pattern of the mSMGP/mPIP gene expression, in addition to our recent demonstration that mSMGP/mPIP is found in mouse saliva and can bind bacteria, suggest that this protein may have a protective role in the mouse.


The possible mechanism of silver nanoparticle impact on hippocampal synaptic plasticity and spatial cognition in rats.
Silver nanoparticles (Ag-np) are very promising engineered nanomaterials which play an important role in the world biomedical, healthcare and in general nanotechnology applications. With the most impressive effect in antibacterial and many other broad-spectrum biotechnological advantages, Ag-np in real applications is still a controversial issue. This study investigated effects of the Ag-np on hippocampal synaptic plasticity and spatial cognition in rats and followed with the research on their possible mechanism. In this study, twenty-four adult male Wister rats were randomly divided into 3 groups: control group, low-dose group (Ag-np, 3 mg/kg) and high-dose group (Ag-np, 30 mg/kg). After two-week exposure to Ag-np through the nasal administration, Morris water maze (MWM) test was performed for the spatial cognition, followed by the long-term potentiation (LTP) recording and reactive oxygen species (ROS) detection in hippocampal homogenate. Results showed that compared with the control group, both LTP and MWM were abnormal in low-dose group and high-dose group. The quantity of ROS in hippocampal homogenate was increased significantly in low-dose group and high-dose group, which may be the reason of the neural damage caused by Ag-np.


The Sexuality of Sex Workers: Sexual Arousability and the Sexual Profile of Women Engaged in Prostitution in Serbia.
In a view of sex work being a sexual behavioural activity, the main objective of our research was to explore the sexuality of sex workers in Serbia regarding to intensity of their sexual arousability. The research was conducted on 30 sex workers and 30 controls of randomly selected social and demographic characteristics, without any signs of psychiatric morbidity. For the evaluation of the intensity of sexual arousability, SAI (Sexual Arousability Index) questionnaire was used. The analysis of the main total scores of the SAI for the sex workers and control participants showed no statistically significant difference in the overall intensity of arousability (p>0.05). However, item by item analysis of the questionnaire showed qualitative difference in sexuality between sex workers and controls, related to certain aspects of sexuality. These findings allow the space for further research in the way of identifying origins of qualitative issues in the sexual profile of sex workers, in correlation to women who are not sex workers, i.e. whether they had been result of potential biological, specific psychodynamic factors, or have been formed as a result of the direct influence of the sex work.


[In vitro study of the stability of reduced intraerythrocyte glutathione against a group of drugs frequently used during the neonatal period].
Authors have studied in 32 samples of cord blood (normal newborns and normal births) the stability of glutathione reduced form (GSH) in human erythrocytes opposite to ten drugs (gentamicin, cephazolin, amikacin, carbenicillin, amoxicillin, benzodiacepine, metil-prednisolone, phenilbarbituric acid, fosfomycin, trimethroprim-sulphamethoxazole) using it at similar concentration to the highest haemotic levels that are obtained in the newborn with therapeutic doses. They showed that these drugs have not an effect on stability of GSH. For this reason is very unprobable that these drugs are cause of haemolitic crisis in the newborn. In the another hand, they suggest the use co-trimoxazole in the newborn always, in correct doses.


Fish proglucagon genes have differing coding potential.
Regulation of tissue-specific hormone production involves transcription, post-translational and physiological regulation. The proglucagon gene has been an ideal molecule for understanding many aspects of regulation of hormone production. Gene duplications often allow the evolution of new gene function, and the proglucagon gene has been duplicated on the lineage leading to teleost fish, while most other vertebrates have only a single proglucagon gene. The recent characterization of near complete pufferfish and zebrafish genomes allowed us to determine the structure of the duplicated genes. We searched the pufferfish and zebrafish genomes for proglucagon-like sequences. Our searches identified two proglucagon-like genes in both the pufferfish and zebrafish genomes. In pufferfish the proglucagon-like genes are surrounded by genes that are similar to genes that flank the proglucagon gene in mammalian genomes, indicating that gene order near the proglucagon gene has been maintained since the fish-mammal divergence approximately 450 million years ago. Characterization of the duplicated fish proglucagon genes indicates that one of the two genes is predicted to encode glucagon, GLP-1 and GLP-2. In contrast, the second proglucagon gene is predicted to encode only glucagon and GLP-1, but not GLP-2. Thus the two fish proglucagon genes have different coding potential, and, therefore likely differ in function. Regulation of gene expression potentially has a role in the regulation of production of proglucagon derived peptides in fish.


Production of ochratoxin a, citrinin, and ergosterol byPenicillium viridicatum in autoclaved and non-autoclaved wheat at low temperature.
Wheat (moisture content: 26%) was autoclaved or left untreated, inoculated with conidia ofPenicillium viridicatum and stored at 10°C. The fungus grew on both substrates and was the dominant mould on the non-autoclaved grain. Autoclaving resulted in an earlier onset of ergosterol, ochratoxin A, and citrinin production due to accelerated mould growth. Yield of ochratoxin A increased while citrinin slightly decreased in autoclaved wheat.


"Older people tend to be invisible": a qualitative study exploring the needs and inclusion of older Syrian refugees in the context of compounding crises in host country, Lebanon.
Older Syrian refugees in Lebanon are a marginalized population with under-recognized health needs. The inclusivity of this population within the humanitarian response is poorly understood. This study aims to identify the unique needs of older Syrian refugees in the context of recent concurrent crises in Lebanon, and explore the extent to which they are being met and prioritized by local and international aid agencies. We conducted in-depth interviews with a snowball sample of 26 stakeholders from 11 organizations operating in the health, nutrition, and water, sanitation, and hygiene sectors. Data analysis followed principles of thematic analysis. Concurrent political, economic, and public health crises in host country promoted income insecurity among older refugees and increased dependency on younger relatives, leading to food insecurity, neglect, and poor health outcomes, including the sequelae of untreated non-communicable diseases. Mental illness was perceived to be exacerbated by Covid-19 related challenges, including social isolation, uncertainty about the future, and additionally due to feelings of guilt related to economic dependence and fundamental exclusion from labor force participation. Despite their vulnerability, older refugees are overlooked by the humanitarian response, which may be related to a lack of data. Pervasive medication shortages in the setting of the economic collapse, as well as inaccessible physical environments and competing interests were all identified as major barriers to care. Older Syrian refugees in Lebanon experience dual vulnerability that is acutely exacerbated in the setting of concurrent crises. Sociopolitical, economic, and cultural barriers promote social exclusion and may confer an increased risk of income and food insecurity in this population, with significant implications for health. Humanitarian aid agencies operating in the context of fragmented, under-resourced health systems are currently unable to sufficiently address multi-faceted needs of this community. We recommend moving away from a donor-dependent model of aid by allocating resources toward strengthening inclusive national health systems that emphasize preventative care. We further call for age-disaggregation of routine data and normalization of data sharing among stakeholders in the academic and public health sectors in order to develop evidence-based initiatives that can meet the needs of this under-served community.


Fabrication of oxidized bacterial cellulose by nitrogen dioxide in chloroform/cyclohexane as a highly loaded drug carrier for sustained release of cisplatin.
Carboxylated bacterial cellulose (OBC) was fabricated by oxidation with nitrogen dioxide in chloroform/cyclohexane and employed as a carrier for sustained release of antitumor substance cisplatin (CDDP). The influence of removing water method, solvent used in the synthesis, concentration of N2O4, and duration of the oxidation on content of carboxyl groups in reaction products was established. Due to the possibility of nitrogen dioxide to penetrate into cellulose crystallites, the carboxyl group content of the OBC reaches high values up to 4 mmol/g. In vitro degradation of OBC was determined under simulated physiological conditions. The immobilization of CDDP on OBC was studied in detail. The initial burst release of the drug from the polymer was depressed. The cytotoxicity of CDDP-loaded OBC was evaluated with HeLa cells. The unique structure and properties of OBC make it a great candidate as drug delivery carrier.


The traumatic pneumomyelogram. A previously undescribed entity.
Air within the cervical subarachnoid space secondary to skull trauma (traumatic air myelogram or pneumomyelogram) has not been reported previously. Such a case is presented and the findings are reviewed in the light of existing literature.


Validation of a model to predict survival in elderly patients with acute myeloid leukaemia.
Intensive chemotherapy is the treatment of choice for selected elderly patients with acute myeloid leukaemia (AML). We recently developed a model to predict survival, thereby providing objective information upon which to select appropriate therapy for such patients. Such models, however, must be validated on a cohort of patients not used during the development of the model. We have tested the model using a series of 61 elderly patients consecutively treated with intensive chemotherapy. Using several statistical techniques, we have shown that the model is of value in predicting prognosis, though two patients did markedly better than the model prediction. This model may be useful for predicting survival in elderly patients with AML and warrants more extensive validation.


Norovirus infection among children with acute gastroenteritis in Recife, Brazil: disease severity is comparable to rotavirus gastroenteritis.
Norovirus has captured increasing attention as an agent of childhood diarrhoea. However, it is not known whether norovirus causes as severe diarrhoea as rotavirus, particularly among children in developing countries. In a 1-year study conducted between May 2004 and April 2005 in Recife, Brazil, norovirus was detected by ELISA in 34/233 (15%) diarrhoeal children less than 5 years of age. The severity of clinical illness, as indicated by the presence of dehydration, the requirement for hospitalization, and the duration of hospital stay, was similar between children with norovirus and rotavirus infection. These data underscore the importance of norovirus as a cause of severe diarrhoea in children.


Connexin channel modulators and their mechanisms of action.
Gap junction channels and hemichannels formed by the connexin family of proteins play important roles in many aspects of tissue homeostasis in the brain and in other organs. In addition, connexin channels have been proposed as pharmacological targets in the treatment of a number of human disorders. In this review, we describe the connexin-subtype selectivity and specificity of pharmacological agents that are commonly used to modulate connexin channels. We also highlight recent progress made toward identifying new agents for connexin channels that act in a selective and specific manner. Finally, we discuss developing insights into possible mechanisms by which these agents modulate connexin channel function. This article is part of the Special Issue Section entitled 'Current Pharmacology of Gap Junction Channels and Hemichannels'.


Mechanism of the protective action of thiol compounds in ethanol-induced liver injury.
The protective action of cysteine or mercaptopropionylglycine (MPG) in acute ethanol-induced liver injury has been investigated in the rat. Cysteine accelerated clearance of ethanol and acetaldehyde from blood and liver and prevented an increase in hepatic content of triglyceride and serum ornithine carbamoyl transferase activity. MPG accelerated clearance of ethanol and acetaldehyde less efficiently but prevented an increase in these variables to the same degree. The mode of action of thiol compounds in acute ethanol-induced liver injury has been discussed.


Comparison of Intraductal RFA Plus Stent versus Stent-Only Treatment for Unresectable Perihilar Cholangiocarcinoma-A Systematic Review and Meta-Analysis.
One of the cornerstones of palliative treatment for unresectable perihilar cholangiocarcinoma is biliary stent placement in order to restore biliary drainage. In this review, the potential added value of RFA with stent placement in comparison to stent placement alone in patients with unresectable perihilar cholangiocarcinoma is analyzed. We performed a comprehensive online search for relevant articles in November 2021 (PROSPERO ID: CRD42021288180). The primary endpoint was difference in overall survival. Secondary endpoints included overall survival, stent patency and complications. Only studies comparing survival after RFA + stent placement with stent placement alone were included in the meta-analysis. Non-comparative studies or comparative studies describing stent patency only were included in the systematic review. A total of nine studies, including 217 patients with pCCA who underwent RFA + stent placement and 294 patients who underwent stent-only treatment, met the inclusion criteria for the primary endpoint analysis. Direct comparison between the two treatment groups showed a significantly longer overall survival for RFA + stent treatment, with a pooled HR of 0.65 [95% CI, 0.50-0.84, I2 = 38%]. When all eligible studies were included, RFA + stent treatment revealed an overall survival of 9.5 months [95% CI, 6.3-12.6], whereas survival for stent-only treatment was 7.0 months [95% CI, 5.7-8.2]. Due to the heterogeneity of the data, no pooled data analysis could be performed on stent patency or complications. RFA + stent placement displays promising potential to prolong survival. However, further research incorporating confounding factors like use of palliative chemotherapy is necessary in order to validate these findings.


Spatial investigation of the elemental distribution in Wilson's disease liver after d-penicillamine treatment by LA-ICP-MS.
At present, the copper chelator d-penicillamine (DPA) is the first-line therapy of Wilson's disease (WD), which is characterized by an excessive copper overload. Lifelong DPA treatments aim to reduce the amount of detrimental excess copper retention in the liver and other organs. Although DPA shows beneficial effect in many patients, it may cause severe adverse effects. Despite several years of copper chelation therapy, discontinuation of DPA therapy can be linked to a rapidly progressing liver failure, indicating a high residual liver copper load. In order to investigate the spatial distribution of remaining copper and additional elements, such as zinc and iron, in rat and human liver samples after DPA treatment, a high resolution (spotsize of 10μm) laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) imaging method was applied. Untreated LPP-/- rats, an established animal model for WD, appeared with a high overall copper concentration and a copper distribution of hotspots distributed over the liver tissue. In contrast, a low (>2-fold decreased) overall copper concentration was detected in liver of DPA treated animals. Importantly, however, copper distribution was highly inhomogeneous with lowest concentrations in direct proximity to blood vessels, as observed using novel zonal analysis. A human liver needle biopsy of a DPA treated WD patient substantiated the finding of an inhomogeneous copper deposition upon chelation therapy. In contrast, comparatively homogenous distributions of zinc and iron were observed. Our study indicates that a high resolution LA-ICP-MS analysis of liver samples is excellently suited to follow efficacy of chelator therapy in WD patients.


Effect of experimentally induced generalized seizures on the wakefulness--sleep cycle.
The effect of experimentally induced generalized seizures on the relations between the various phases of sleep in the wakefulness--sleep cycle was studied in cats with chronically implanted electrodes. After generalized convulsions induced by electrical stimulation of the dorsal hippocampus, unlike those of amygdalar origin, clearly defined changes were observed in the structure of the wakefulness--sleep cycle. In the postseizure period, with increasing wakefulness sharp depression of paradoxical sleep takes place. However, in slow wave sleep only slight changes were observed. Instead of a rebound phenomenon, paradoxical sleep was sharply induced in cats after preliminary deprivation of paradoxical sleep as a result of generalized seizures induced by electrical stimulation of the neocortex. Nonspecific hyperactivation of the brain, in the form of epileptiform discharges, thus has a particularly marked effect on the structure of paradoxical sleep.


Family structure, social capital, and mental health disparities among Canadian mothers.
To examine the extent to which inequities in mental health between single and partnered mothers can be explained by social capital, independently and in concert with socio-economic circumstances. Cross-sectional study. The sample consisted of 2920 mothers participating in Statistics Canada's 2010 General Social Survey. Chi-square and logistic regression analyses were used to investigate the extent to which family structure differences in self-rated mental health, if observed, were mediated by various dimensions of social capital. Compared with partnered mothers, fair/poor self-rated mental health was more common among previously married mothers (OR = 3.14; 95% CI 2.15-4.59) and never married mothers (OR = 3.01; 95% CI 1.95-4.65). After adjustment for socio-economic and social capital variables, the odds ratio between single mother family structure and fair/poor mental health decreased but remained significant (ORpreviously married = 1.90, 95% CI 1.22-2.98; ORnever married = 1.90, 95% CI 1.14-3.16). Single mothers' more limited access to economic and social capital resources partially explain their compromised self-rated mental health. Longitudinal research with multi-item measures of mental health is needed to corroborate these findings and extend their understanding of the relationship between family structure, social capital, and mothers' mental health.


Synthesis of ticagrelor analogues belonging to 1,2,3-triazolo[4,5-d]pyrimidines and study of their antiplatelet and antibacterial activity.
Based on the recent observation that the antiplatelet agent ticagrelor and one of its metabolite exert bactericidal activity against gram-positive bacteria, a series of 1,2,3-triazolo[4,5-d]pyrimidines structurally related to ticagrelor were synthesized and examined as putative antiplatelet and antibacterial agents. The aim was to assess the possibility of dissociating the two biological properties and to find novel 1,2,3-triazolo[4,5-d]pyrimidines expressing antiplatelet activity and devoid of in vitro antibacterial activity. The new compounds synthesized were known metabolites of ticagrelor as well as structurally simplified analogues. Some of them were found to express antiplatelet activity and to lose the antibacterial activity, supporting the view that the two activities were not necessarily linked.


The Effects of the Binder and Buffering Matrix on InSb-Based Anodes for High-Performance Rechargeable Li-Ion Batteries.
C-decorated intermetallic InSb (InSb-C) was developed as a novel high-performance anode material for lithium-ion batteries (LIBs). InSb nanoparticles synthesized via a mechanochemical reaction were characterized using X-ray diffraction (XRD), high-resolution transmission electron microscopy (HRTEM), scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS), and energy-dispersive X-ray spectroscopy (EDX). The effects of the binder and buffering matrix on the active InSb were investigated. Poly(acrylic acid) (PAA) was found to significantly improve the cycling stability owing to its strong hydrogen bonding. The addition of amorphous C to InSb further enhanced mechanical stability and electronic conductivity. As a result, InSb-C demonstrated good electrochemical Li-ion storage performance: a high reversible specific capacity (878 mAh·g-1 at 100 mA·g-1 after 140 cycles) and good rate capability (capacity retention of 98% at 10 A·g-1 as compared to 0.1 A·g-1). The effects of PAA and C were comprehensively studied using cyclic voltammetry, differential capacity plots, ex-situ SEM, and electrochemical impedance spectroscopy (EIS). In addition, the electrochemical reaction mechanism of InSb was revealed using ex-situ XRD. InSb-C exhibited a better performance than many recently reported Sb-based electrodes; thus, it can be considered as a potential anode material in LIBs.


Toll-like receptor 2 mediates persistent chemokine release by Chlamydia pneumoniae-infected vascular smooth muscle cells.
The intracellular bacterium Chlamydia pneumoniae is present in many atherosclerotic lesions, where it could promote inflammation. This study determined whether monocyte chemoattractant protein 1 (MCP-1) release is stimulated in vascular smooth muscle cells (VSMCs) that are exposed to or infected by C pneumoniae and whether toll-like receptor 2 (TLR2) or TLR4 mediate these effects. TLR2 mRNA was expressed constitutively and was upregulated by C pneumoniae exposure in mouse aortic SMC and was inducible by C pneumoniae and TLR3 and TLR4 agonists in human coronary artery SMCs. Exposure to inactivated or viable extracellular C pneumoniae evoked a robust increase in MCP-1 release and activated nuclear factor-kappaB and extracellular signal-regulated kinase 1/2 in wild-type and TLR4 signaling-deficient mouse aortic SMCs but not in TLR2-deficient SMCs, probably because of TLR2-mediated recognition of a chlamydial antigen. Brief exposure to viable C pneumoniae led to active infection of VSMCs, shown by chlamydial protein synthesis, and caused a persistent (>48-hour) MCP-1 release that was also TLR2 dependent. The results show that VSMCs express functional TLR2 and that TLR2 mediates both a persistent activation of chemokine release in C pneumoniae-infected VSMCs and its acute stimulation by extracellular C pneumoniae. Therefore, TLR2 expressed in VSMCs may promote inflammation within the arterial wall.


Characterization of TRPM8-expressing neurons in the adult mouse hypothalamus.
Transient receptor potential melastatin 8 (TRPM8) is a menthol receptor that detects cold temperatures and influences behaviors and autonomic functions under cold stimuli. Despite the well-documented peripheral roles of TRPM8, the evaluation of its central functions is still of great interest. The present study clarifies the nature of a subpopulation of TRPM8-expressing neurons in the adult mice. Combined in situ hybridization and immunohistochemistry revealed that TRPM8-expressing neurons are exclusively positive for glutamate decarboxylase 67 mRNA signals in the lateral septal nucleus (LS) and preoptic area (POA) but produced no positive signal for vesicular glutamate transporter 2. Double labeling immunohistochemistry showed the colocalization of TRPM8 with vesicular GABA transporter at axonal terminals. Immunohistochemistry further revealed that TRPM8-expressing neurons frequently expressed calbindin and calretinin in the LS, but not in the POA. TRPM8-expressing neurons in the POA expressed a prostaglandin E2 receptor, EP3, and neurotensin, whereas expression in the LS was minimal. These results indicate that hypothalamic TRPM8-expressing neurons are inhibitory GABAergic, while the expression profile of calcium-binding proteins, neurotensin, and EP3 differs between the POA and LS.


Sodium ferulate prevents amyloid-beta-induced neurotoxicity through suppression of p38 MAPK and upregulation of ERK-1/2 and Akt/protein kinase B in rat hippocampus.
To observe whether an amyloid beta (Abeta)-induced increase in interleukin (IL)-1beta was accompanied by an increase in the p38 mitogen-activated protein kinase (MAPK) pathway and a decrease in the cell survival pathway, and whether sodium ferulate (SF) treatment was effective in preventing these Abeta-induced changes. Rats were injected intracerebroventricularly with Abeta25-35. Seven days after injection, immunohistochemical techniques for glial fibrillary acidic protein (GFAP) were used to determine the astrocyte infiltration and activation in hippocampal CA1 areas. The expression of IL-1beta, extracellular signal-regulated kinase (ERK), p38 MAPK, Akt/protein kinase B (PKB), Fas ligand and caspase-3 were determined by Western blotting. The caspase-3 activity was measured by cleavage of the caspase-3 substrate (Ac-DEVD-pNA). Reverse transcription-polymerase chain reaction was used to analyze the changes in IL-1beta mRNA levels. Intracerebroventricular injection of Abeta25-35 elicited astrocyte activation and infiltration and caused a strong inflammatory reaction characterized by increased IL-1beta production and elevated levels of IL-1beta mRNA. Increased IL-1beta synthesis was accompanied by increased activation of p38 MAPK and downregulation of phospho-ERK and phospho-Akt/PKB in hippocampal CA regions prepared from Abeta-treated rats, leading to cell death as assessed by activation of caspase-3. SF significantly prevented Abeta-induced increases in IL-1beta and p38 MAPK activation and also Abeta-induced changes in phospho-ERK and phospho-Akt/PKB expression levels. SF prevents Abeta-induced neurotoxicity through suppression of p38 MAPK activation and upregulation of phospho-ERK and phospho-Akt/PKB expression.


Investigation of the interaction between Genetic Risk Score (GRS) and fatty acid quality indices on mental health among overweight and obese women.
Mental disorders are associated with dietary fatty acids and genome-wide association studies have found multiple risk loci robustly related to depression, anxiety, and stress. The aim of this study is to investigate the interaction of genetic risk score (GRS) and dietary fat quality indices on mental health. This cross-sectional study included 279 overweight and obese women for N6/N3 ratio and 378 overweight and obese women for CSI aged 18-68 years. Using reliable and verified standard protocols, body composition, anthropometric indices, blood pressure, physical activity, and dietary fat quality were measured. Serum samples were used to determine biochemical tests. A genetic risk score (GRS) was calculated using the risk alleles of the three SNPs. A generalized linear model (GLM) was applied to assess the interactions between GRS and fat quality indices. Mental health was evaluated using Depression Anxiety Stress Scales (DASS-21). The mean (± SD) age and BMI of our participants were 36.48 (8.45) and 30.73 (3.72) kg/m2 respectively. There was a marginally significant mean difference among tertiles of the CSI in terms of stress (P = 0.051), DASS-21 (P = 0.078) in the crude model. After adjusting for age, energy intake, physical activity and BMI in model 1, there was a positive interaction between GRS and T3 of N6/N3 ratio on anxiety (β = 0.91, CI = 0.08,1.75, P = 0.031), depression (β = 1.05, CI = 0.06,2.04, P = 0.037), DASS-21 (β = 2.22, CI= -0.31,4.75, P = 0.086). Our findings indicate that higher ratio of N-6 to N-3 considering genetics were predictive of mental disorder in our population.


Biochar benefits carbon off-setting in blue-green infrastructure soils - A lysimeter study.
Carbon sequestration with amendments in blue-green infrastructure soils could off-set anthropogenic greenhouse gas emissions to alleviate climate change. In this 3-year study, the effects of wheat straw and its biochar on carbon sequestration in an urban landscaping soil were investigated under realistic outdoor conditions using two large-scale lysimeters. Both amendments were carried out by incorporating pellets at 0-15 cm soil depth with an equivalent initial total carbon input of 2% of the dry soil weight. Soil carbon, carbon isotope ratios, dissolved carbon in leachates, CO2-C emissions, carbon fixed in above ground vegetation, soil water content, soil bulk electrical conductivity, and water infiltration rates, were then compared between the 2 lysimeters. After 3 years, we observed that, despite having a 17.2% lower vegetation growth, soil organic and inorganic carbon content was higher by 28.8% and 41.5%, respectively, in biochar as compared to wheat straw amended soil. Carbon isotope analysis confirmed the greater stability of the added carbon in the biochar amended soil. Water content was on average 23.2% and 13.0% in the straw pellet and biochar amended soil, respectively, whereas water infiltration rates were not significantly different between the two lysimeters. Overall, the incorporation of wheat straw biochar into soil could store an estimated 30 tonnes of carbon per hectare in city blue-green infrastructure spaces. Interviews involving institution stakeholders examined the feasibility of this biochar application. Stakeholders recognized the potential of biochar as an environment-friendly means for carbon offsetting, but were concerned about the practicality of biochar production and application into soil and increased maintenance work. Consequently, additional potential benefits of biochar for environmental management such as improving the quality of polluted run-off in stormwater treatment systems should be emphasized to make biochar an attractive proposition in sustainable urban development.


Brain Selective Estrogen Treatment Protects Dopaminergic Neurons and Preserves Behavioral Function in MPTP-induced Mouse Model of Parkinson's Disease.
Parkinson's disease (PD) is characterized by progressive degeneration of dopaminergic neurons in the substantia nigra and loss of both motor and non-motor features. Several clinical and preclinical studies have provided evidence that estrogen therapy reduces the risk of PD but have limitations in terms of adverse peripheral effects. Therefore, we examined the potential beneficial effects of the brain-selective estrogen prodrug, 10β, 17β-dihydroxyestra-1,4-dien-3-one (DHED) on nigrostriatal dopaminergic neurodegeneration and behavioral abnormalities in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mouse model of PD. Wild-type mice were treated with daily subcutaneous injections of DHED (50 and 100 µg/kg) or vehicle for four weeks. To produce PD-like symptoms, mice were injected with MPTP (18 mg/kg in saline; intraperitoneally) four times at 2-hr intervals for one day. After behavioral examination, mice were sacrificed, and the brains were isolated for neurochemical and morphological examinations. MPTP injected mice exhibited loss of dopaminergic neurons and fibers in substantia nigra and striatum respectively, along with impaired motor function at day 7 post MPTP injection. These phenotypes were associated with significantly increased oxidative stress and inflammatory responses in the striatum regions. DHED treatments significantly mitigated behavioral impairments and dopaminergic neurodegeneration induced by MPTP. We further observed that DHED treatment suppressed oxidative stress and inflammation in the striatum of MPTP treated mice when compared to vehicle treated mice. In conclusions, our findings suggest that DHED protects dopaminergic neurons from MPTP toxicity in mouse model of PD and support a beneficial effect of brain-selective estrogen in attenuating neurodegeneration and motor symptoms in PD-related neurological disorders. Graphical Abstract.


Exploring nursing students' need for social presence and its relevance to their learning preferences.
To explore nursing students' need for social presence in online learning, looking at its relevance to the adopted sensory modality learning styles. A qualitative descriptive design. Two semi-structured online focus groups were held with five second-year and seven third-year undergraduate nursing students, purposively recruited from the Malta College of Arts, Science and Technology. Using a structured guide based on the Social Presence Model, participants were asked about their perceived need for social presence (a construct used to understand online interactions), and its relevance to their learning preferences using the VARK (Visual, Aural/Auditory, Read/write and Kinesthetic) learning styles. Despite varying learning preferences, all participants remarked on the need for social presence. Social presence was required for their perceived learning and for actively participating in online lectures. Several participants expressed the need for social presence for a collaborative constructivist learning experience, drawing on their diverse learning styles for a better learning experience.


Hypertriglyceridemia is a Risk Factor for Fever in Early Acute Non-biliary Pancreatitis.
Fever is a common symptom of acute pancreatitis (AP). This study examined the factors associated with fever due to pancreatic inflammation in the early stages of non-biliary AP. This study analyzed the AP database from Kangwon National University Hospital from January 2018 until April 2021 and identified patients who developed fever within 1 week of hospitalization. Patients with gallstone pancreatitis, pseudocyst, walled-off necrosis, chronic pancreatitis, bacteremia, and other site infections were excluded. The febrile group was compared with the afebrile group. One hundred and fifty-two patients were analyzed, and fever was diagnosed in 79 patients (52.0%). Based on multivariate analysis, fever was positively correlated with hypertriglyceridemia-induced AP (OR 16.8, 95% CI 4.0-70.7, p<0.01) and computed tomography severity index (OR 1.7, 95% CI 1.2-2.6, p<0.01). Recurrent AP was negatively associated with fever (OR 0.3, 95% CI 0.1-0.8, p=0.01). Fever was more frequent in patients with higher initial serum triglyceride (TG) levels than those with lower levels (TG <200 mg/dL; 35.1%, 200≤TG<500 mg/dL; 42.3%, TG ≥500 mg/dL; 88.6%, p<0.01). Hypertriglyceridemia and CT severity index are associated with fever in early non-biliary AP.


Interdomain cytoplasmic interactions govern the intracellular trafficking, gating, and modulation of the Kv2.1 channel.
Voltage-gated potassium (Kv) channels comprise four transmembrane alpha subunits, often associated with cytoplasmic beta subunits that impact channel expression and function. Here, we show that cell surface expression, voltage-dependent activation gating, and phosphorylation-dependent modulation of Kv2.1 are regulated by cytoplasmic N/C interaction within the alpha subunit. Kv2.1 surface expression is greatly reduced by C-terminal truncation. Tailless Kv2.1 channels exhibit altered voltage-dependent gating properties and lack the bulk of the phosphorylation-dependent modulation of channel gating. Remarkably, the soluble C terminus of Kv2.1 associates with tailless channels and rescues their expression, function, and phosphorylation-dependent modulation. Soluble N and C termini of Kv2.1 can also interact directly. We also show that the N/C-terminal interaction in Kv2.1 is governed by a 34 aa motif in the juxtamembrane cytoplasmic C terminus, and a 17 aa motif located in the N terminus at a position equivalent to the beta subunit binding site in other Kv channels. Deletion of either motif disrupts N/C-terminal interaction and surface expression, function, and phosphorylation-dependent modulation of Kv2.1 channels. These findings provide novel insights into intrinsic mechanisms for the regulation of Kv2.1 trafficking, gating, and phosphorylation-dependent modulation through cytoplasmic N/C-terminal interaction, which resembles alpha/beta subunit interaction in other Kv channels.


Clinical and health-related quality-of-life outcomes after early and late spinal fusion in pediatric patients with congenital scoliosis at 10-year follow-up.
A retrospective comparative study. This study compares the effect of age at the time of surgery on clinical and health-related quality-of-life (HRQoL) outcomes at 10-year follow-up in pediatric patients with congenital scoliosis (CS). Several studies have evaluated the outcomes of surgical treatments of CS during the growth period; however, age at surgery and its long-term effects have been assessed in only a few case reports. We enrolled patients with CS who underwent spinal fusion at the age of 18 years or younger in our study. We evaluated 97 patients (38 males, 59 females; average age 16.5 years) who met our inclusion criteria, including the availability of outcome data for a minimum of 10-year post-surgery. We divided patients into two groups in terms of the age at surgery: early fusion (EF) and late fusion (LF) groups. Clinical outcomes included re-operations, major curve corrections immediately and at 10-year follow-up, Scoliosis Research Society (SRS)-22 questionnaire, and percentage forced vital capacity (%FVC). The EF group (33 patients) and the LF group (64 patients) did not differ significantly in terms of demographics. In all domains, the EF group had better HRQoL than the LF group. More patients (52%) in the EF group required re-operation than in the LF group (23%). In addition, patients with short fusion (< 7 segments, p = 0.0011) and greater T1-T12 height (≥ 22 cm, p = 0.0088) had better %FVC than their counterparts. Age at surgery might have some non-negligible impacts on patients' HRQoL and clinical outcomes. Our study highlighted the important factors in surgical considerations of choosing the appropriate timing for spinal fusion, performing shorter fusions, and achieving an acceptable curve correction without allowing further curve progression that required re-operations. Level III.


Analysing mucin degradation.
Turnover of mucins in supramucosal gels is essential for the removal of surface contaminants, and the maintenance of normal mucosal barrier function. In addition to the well-known processes promoting the physical turnover of mucus gels, extracellular mucin degradation also requires the coordinated action of a range of enzyme activities including glycosidases and proteases. These are collectively termed "mucinase". Derangements of mucinase activity lead to downstream barrier defects and mucosal disease. This chapter is focussed on methods that can be used to assess the degradation of whole mucins and isolated mucin glycans. A range of approaches is described using labelled or unlabelled substrates utilised in assays based on 96-well plates, size exclusion chromatography, and NP-HPLC. These are suitable for defining the extent and progress of mucin degradation in different mucosal systems, and identifying abnormalities and critical control points.


Neuromuscular transmission and muscle contractility in SNAP-25-deficient coloboma mice.
Synaptosomal associated protein of 25 kDa (SNAP-25) is a cytoplasmic protein that participates in the docking and fusion of synaptic vesicles with the nerve terminal in preparation for neurotransmitter release. SNAP-25 is also a substrate for three of the seven serotypes of botulinum neurotoxin (BoNT). Intoxication by BoNT/A, /C1 or /E results in weakness and paralysis of skeletal muscle due to cleavage of SNAP-25 (and syntaxin la in the case /C1) at discrete serotype-specific sites. To elucidate the role of SNAP-25 in muscle function in more detail, contractility and neuromuscular transmission were studied in a mutant mouse model termed coloboma. The coloboma mutation results from a contiguous deletion of 1-2 centiMorgans on chromosome 2, which includes the entire SNAP-25 locus and three other identified genes. Homozygotes do not survive beyond gestation day 6; heterozygotes (Cm/+) have a normal life-span but express reduced levels of SNAP-25 mRNA and protein in the brain. The consequences of the Cm/+ mutation on twitch and tetanic tension, quantal release of neurotransmitter and spinal motoneuron expression of SNAP-25 were examined in the present study. Contrary to expectations, Cm/+ mice exhibited no alteration in twitch tension and generated normal tetanic tension even at the highest frequency examined (800 Hz). Microelectrode recordings revealed that MEPP amplitude and frequency were both within control limits. The ventral spinal cord of Cm/+ mice showed no deficiency in SNAP-25 content and immunohistochemical examination of nerve terminals in Cm/+ mice disclosed that SNAP-25 levels and distribution were similar to those of control mice. It is concluded that spinal motor neurons up-regulate SNAP-25 to preserve vital neuromuscular function.


Determination of body water compartments in neonatal foals by use of indicator dilution techniques and multifrequency bioelectrical impedance analysis.
To determine values for total body water (TBW), extracellular fluid volume (ECFV), intracellular fluid volume (ICFV), and plasma volume (PV) in healthy neonatal (< 24 hours old) foals and to create a multifrequency bioelectrical impedance analysis (MF-BIA) model for use in neonatal foals. 7 healthy neonatal foals. Deuterium oxide (0.4 g/kg, IV), sodium bromide (30 mg/kg, IV), and Evans blue dye (1 mg/kg, IV) were administered to each foal. Plasma samples were obtained following an equilibration period, and the TBW, ECFV, ICFV, and PV were calculated for each foal. An MF-BIA model was created by use of morphometric measurements from each foal. Mean ± SD values were obtained for TBW (0.744 ± 0.024 L/kg), ICFV (0.381 ± 0.018 L/kg), ECFV (0.363 ± 0.014 L/kg), and PV (0.096 ± 0.015 L/kg). The 95% limits of agreement between the MF-BIA and indicator dilution techniques were within ± 2 L for TBW and ECFV. Fluid volumes in neonatal foals were found to be substantially larger than fluid volumes in adult horses. Multifrequency bioelectrical impedance analysis may be a useful technique for predicting TBW, ICFV, and ECFV in neonatal foals.


Deficiency of interleukin-1 receptor antagonist promotes spontaneous femoral artery aneurysm formation in mice.
Femoral artery aneurysms (FAAs) are very rare, and their natural history is not well understood. In this study, we sought to analyze the pathogenesis of inflammatory FAAs in interleukin-1 receptor antagonist-deficient (IL-1Ra(-/-)) B6 mice. Systolic arterial pressures and plasma lipid levels of IL-1Ra(-/-) mice and wild-type (WT) mice did not differ significantly. However, IL-1Ra(-/-) mice spontaneously developed fusiform FAAs. Real-time PCR of 9-month-old IL-1Ra(-/-) mice revealed significantly increased mRNA levels of IL-1β (6.6-fold), tumor necrosis factor-α (TNF-α) (12.4-fold), and matrix metalloproteinase-9 (6.0-fold) compared with WT mice. Histological analysis revealed numerous inflammatory cells around the FAAs in IL-1Ra(-/-) mice, and elastin staining showed destruction of both the internal and external elastic lamina in IL-1Ra(-/-) mice. Afterward, macrophage function was studied. After lipopolysaccharide (1 μg/mL) stimulation, IL-1Ra-deficient macrophages produced much higher levels of TNF-α than those from WT mice. Finally, we performed bone marrow cell transplantation. FAAs with many inflammatory cells in the adventitia were detected in several WT mice that received bone marrow cells from IL-1Ra(-/-) mice (44%), but not from WT mice (0%). Our study is the first to demonstrate that IL-1Ra deficiency in inflammatory cells disrupts immune system homeostasis and induces inflammatory FAAs in IL-1Ra(-/-) B6 mice. We believe that these mice will provide much information about the natural history and management of FAAs.