Effects of total flavonoids from Drynariae Rhizoma prevent bone loss in vivo and in vitro.
Estrogen deficiency is one of the major causes of osteoporosis in postmenopausal women. Drynariae Rhizoma is a widely used traditional Chinese medicine for the treatment of bone diseases. In this study, we investigated the therapeutic effects of the total Drynariae Rhizoma flavonoids (DRTF) on estrogen deficiency-induced bone loss using an ovariectomized rat model and osteoblast-like MC3T3-E1 cells. Our results indicated that DRTF produced osteo-protective effects on the ovariectomized rats in terms of bone loss reduction, including decreased levels of bone turnover markers, enhanced biomechanical femur strength and trabecular bone microarchitecture deterioration prevention. In vitro experiments revealed that the actions of DRTF on regulating osteoblastic activities were mediated by the estrogen receptor (ER) dependent pathway. Our data also demonstrated that DRTF inhibited osteoclastogenesis via up-regulating osteoprotegrin (OPG), as well as down-regulating receptor activator of NF-κB ligand (RANKL) expression. In conclusion, this study indicated that DRTF treatment effectively suppressed bone mass loss in an ovariectomized rat model, and in vitro evidence suggested that the effects were exerted through actions on both osteoblasts and osteoclasts.


Nontrivial Topological Surface States in Ru3 Sn7 toward Wide pH-Range Hydrogen Evolution Reaction.
Nontrivial topological surface states (TSSs), which possess extraordinary carrier mobility and are protected by the bulk symmetry, have emerged as an innovative platform to search for efficient electrocatalysts toward hydrogen evolution reaction (HER). Here, a Sn-based nontrivial metal Ru3 Sn7 is prepared using electrical arc melting method. The results indicate that the (001) crystal family of Ru3 Sn7 possesses nontrivial TSSs with linear dispersion relation and large nontrivial energy window. Experimental and theoretical results demonstrate that nontrivial TSSs of Ru3 Sn7 can significantly boost charge transfer kinetics and optimize adsorption of hydrogen intermediates due to bulk symmetry-protected band structures. As expected, nontrivial Ru3 Sn7 exhibits superior HER activity to Ru, Pt/C, and trivial counterparts (e.g., Ru2 Sn3 , IrSn2 , and Rh3 Sn2 ) with higher ratios of noble metals. Furthermore, the wide pH-range activity of topologically nontrivial Ru3 Sn7 implies the robustness of its TSSs against pH variation during the HER. These findings provide a promising approach to the rational design of topologically nontrivial metals as highly efficient electrocatalysts.


Identification of novel chicken CD4⁺ CD3⁻ blood population with NK cell like features.
Chicken NK cells have been defined in embryonic spleen and intestinal epithelium as CD8(+) lymphoid cells that lack BCR and TCR, whereas blood NK cells have not been phenotypically defined. Here we employed the mab, 8D12 directed against CHIR-AB1, a chicken Fc receptor, to define a previously uncharacterized lymphoid cell population in the blood. Although CHIR-AB1 expression was found on several cell populations, cells with extraordinary high CHIR-AB1 levels ranged between 0.4 and 2.8% in five different chicken lines. The widespread applicability of the CHIR-AB1 mab was unexpected, since CHIR-AB1-like genes form a polygenic and polymorphic subfamily. Surprisingly the CHIR-AB1 high cells coexpressed low MHCII, low CD4 and CD5, while other T cell markers CD3 and CD8, the B cell marker Bu1, the macrophage marker KUL01 were absent. Moreover, they stained with the mab 28-4, 20E5 and 1G7, which define chicken NK cells and they also expressed CD25, CD57, CD244 and the vitronectin receptor (αVβ3 integrin). In functional assays, PMA stimulation led to high levels of IFNγ release, while spontaneous cytotoxicity was not detectable. The expression of typical NK cell markers in the absence of characteristic B- or T-cell markers, and their IFNγ release is suggestive of a yet unidentified NK like population.


Adaptive sample size determination for the development of clinical prediction models.
We suggest an adaptive sample size calculation method for developing clinical prediction models, in which model performance is monitored sequentially as new data comes in. We illustrate the approach using data for the diagnosis of ovarian cancer (n = 5914, 33% event fraction) and obstructive coronary artery disease (CAD; n = 4888, 44% event fraction). We used logistic regression to develop a prediction model consisting only of a priori selected predictors and assumed linear relations for continuous predictors. We mimicked prospective patient recruitment by developing the model on 100 randomly selected patients, and we used bootstrapping to internally validate the model. We sequentially added 50 random new patients until we reached a sample size of 3000 and re-estimated model performance at each step. We examined the required sample size for satisfying the following stopping rule: obtaining a calibration slope ≥ 0.9 and optimism in the c-statistic (or AUC) < = 0.02 at two consecutive sample sizes. This procedure was repeated 500 times. We also investigated the impact of alternative modeling strategies: modeling nonlinear relations for continuous predictors and correcting for bias on the model estimates (Firth's correction). Better discrimination was achieved in the ovarian cancer data (c-statistic 0.9 with 7 predictors) than in the CAD data (c-statistic 0.7 with 11 predictors). Adequate calibration and limited optimism in discrimination was achieved after a median of 450 patients (interquartile range 450-500) for the ovarian cancer data (22 events per parameter (EPP), 20-24) and 850 patients (750-900) for the CAD data (33 EPP, 30-35). A stricter criterion, requiring AUC optimism < = 0.01, was met with a median of 500 (23 EPP) and 1500 (59 EPP) patients, respectively. These sample sizes were much higher than the well-known 10 EPP rule of thumb and slightly higher than a recently published fixed sample size calculation method by Riley et al. Higher sample sizes were required when nonlinear relationships were modeled, and lower sample sizes when Firth's correction was used. Adaptive sample size determination can be a useful supplement to fixed a priori sample size calculations, because it allows to tailor the sample size to the specific prediction modeling context in a dynamic fashion.


Efficacy and safety of general anesthesia deep brain stimulation for dystonia: an individual patient data meta-analysis of 341 cases.
The efficacy and safety of deep brain stimulation (DBS) under general anesthesia for the treatment of dystonia have not yet been confirmed with high level of evidence. This meta-analysis with pooled individual patient data aims to assess the clinical outcomes and identify the potential prognostic factors of dystonia patients who underwent general anesthesia DBS. We searched PubMed, Web of Science, and Embase for articles describing patients with dystonia who underwent asleep DBS and had individual Burke-Fahn-Marsden Dystonia Rating Scale (BFMDRS) scores. The relative improvement in BFMDRS scores was considered the primary outcome. Pearson correlation analyses and multivariate linear regression analysis were conducted to explore the prognostic factors. A total of 34 studies involving 341 patients were included. The mean postoperative improvement in BFMDRS-M (BFMDRS movement subscale) and BFMDRS-D (BFMDRS disability subscale) scores were 58.6±36.2% and 48.5±38.7% at the last follow-up visit, respectively, with a mean follow-up time of 22.4±27.6 months. Age at surgery and disease duration showed a negative correlation with the percent improvement of BFMDRS-M (%) at the last visit (r=-0.134, P=0.013; r=-0.165, P=0.006). In the stepwise multivariate regression, only disease duration remained a relevant factor. Additionally, the adverse events were acceptable. General anesthesia DBS is a safe, effective, and feasible option for dystonia patients in the long term. Shorter disease duration predicts better clinical outcomes.


HIV testing trends among gay men in Scotland, UK (1996-2005): implications for HIV testing policies and prevention.
To examine trends in the HIV testing behaviour of gay men in Scotland over a 10-year period. Seven cross-sectional surveys in commercial gay venues in Glasgow and Edinburgh (1996-2005). 9613 men completed anonymous, self-completed questionnaires (70% average response rate). Among 8305 respondents included in these analyses, HIV testing increased between 1996 and 2005, from 49.7% to 57.8% (p<0.001). The proportion of men who had tested recently (in the calendar year of, or immediately before, the survey) increased from 28.4% in 1996 to 33.2% in 2005, when compared with those who have tested but not recently, and those who have never tested (adjusted odds ratio 1.31, 95% CI 1.13 to 1.52). However, among ever testers, there was no increase in rates of recent testing. Recent testing decreased with age: 31.3% of the under 25, 30.3% of the 25-34, 23.2% of the 35-44 and 21.2% of the over 44 years age groups had tested recently. Among men reporting two or more unprotected anal intercourse partners in the previous year, only 41.4% had tested recently. HIV testing among gay men in Scotland increased between 1996 and 2005, and corresponds with the Scottish Government policy change to routine, opt-out testing in genitourinary medicine clinics. Testing rates remain low and compare unfavourably with near-universal testing levels elsewhere. The limited change and decline across age groups in recent HIV testing rates suggest few men test repeatedly or regularly. Additional, innovative efforts are required to increase the uptake of regular HIV testing among gay men.


Anti-mu treatment suppresses immunoglobulin light-chain gene expression and Peyer's patch development.
Anti-mu treated mice have been used extensively as a model for suppressed B-cell development [Murgita R. A., Mattioli C. A. and Tomasi T. B., Jr. (1973) J. exp. Med. 138, 209; Manning D. D. (1975) J. Reticuloendothel. Soc. 18, 63; Manning D. D. and Jutila J. W. (1972) J. Immun. 108, 282; Janeway C. A., Jr., Murgita R. A., Weinbaum F. I., Asofsky R. and Wigzell H. (1977) Proc. natn. Acad. Sci. U.S.A. 74, 4582; Hayglass K. T., Naides S. J., Benacerraf B. and Sy M.-S. (1985) J. Molec. Cell. Immun. 2, 107; Manning D. D. (1972) J. Immun. 109, 1152; Cooper M. D., Kearney J. F., Gathings W. E. and Lawton A. R. (1980) Immun. Rev. 52, 29; Burrows P. D., Kearney J. F., Lawton A. R. and Cooper M. D. (1978) J. Immun. 120, 1526]. However, little molecular evaluation has been performed on these animals to determine the level at which B-lineage cells are arrested. Experiments reported here were designed to determine the effects of anti-mu treatment of newborn mice on Ig-specific mRNA expression in lymphocyte populations. Newborn CBA/J mice received i.p. injections of goat anti-mu IgG or non-immune goat IgG, every 2 days, from birth until age 4 weeks. The degree of B-cell suppression in anti-mu treated mice was evident by low serum Ig levels and lack of surface Ig+ cells in splenic lymphocytes. Morphologically, spleens of B-cell depleted mice were slightly reduced or normal size, while the total area of Peyer's patches (PP) was three-fold less than control mice. Spleen cells from anti-mu suppressed mice contained high levels of mu-mRNA, but markedly reduced levels of mRNA specific for other Ig heavy-chain isotypes, as determined by DNA excess dot blot and Northern blot hybridizations. RNA specific for other sequences (actin or IL-2 receptor) was not affected and hybridization to parent plasmid (pACYC) was not detected. In addition, suppression of kappa- and lambda-mRNA accumulation was evident. This was surprising, since the target for anti-mu treatment appears to be a B-cell population expressing intact surface IgM, a stage in B-cell development in which both mu- and light-chain-specific mRNA accumulation should be detected. Our results suggest one of the following models: (1) anti-mu treatment deletes all Ig+ cells from the animal, so that only mu expressing pre-B-cells remain; or (2) anti-mu suppresses B-cell development by inhibiting kappa and lambda transcription, perhaps by some feedback mechanism in which the presence of surface Ig is required to maintain light-chain transcription.


The diagnostic value of haematuria and proteinuria in Schistosoma haematobium infection in southern Nigeria.
Haematuria and proteinuria as detected by chemical reagent strips correlated moderately (r = 0.7) with prevalence and intensity of infection with Schistosoma haematobium in an area of Anambra State, Nigeria. Differences attributable to age and sex were also reflected in a similar pattern, all peaks occurring in the 5-14 year age group. The differences observed with varying levels of intensity and haematuria at both 10 and 50 erythrocytes/microliter (p < 0.001) and proteinuria at 0.3 g/dl (p < 0.01) were statistically significant. At a proteinuria level of 1 g/dl, the observed differences were however not statistically significant (p > 0.5). The percentage of specimens from children (0-14 years) positive for S. haematobium eggs and with at least traces of haematuria and proteinuria (63.4% and 95%, respectively) was higher than in adults (33.3% and 80.2%, respectively). All individuals with more than 50 eggs/10 ml of urine were correctly identified using both indices either separately or in combination. For egg counts of less than 50 eggs/10 ml of urine, false diagnosis occurred in only 5% of all specimens examined. The sensitivity and specificity of haematuria and proteinuria at trace quantities was very high, but haematuria had a higher predictive value for a positive test (PvPt) and was considered the overall better indicator. A combination of both indices did not significantly increase the PvPt. When trace haematuria and moderate proteinuria were combined, both the sensitivity, specificity and PvPt were all above 90%, giving the best overall values in all the combinations made.


Simulation of the Movement of a Locust Swarm in Argentina in 2020.
This work presents a simulation for the trajectory of a locust swarm of the species Schistocerca cancellata (Audinet-Serville), between May 22nd and July 29th, 2020, in Argentina. To obtain the directions, temperature, and intensities of the wind, used to determine the daily traveled distance of the insects, the data of weather forecast from Weather Research and Forecasting (WRF) model are used. A statistical analysis shows the effectiveness of the forecast model used in comparison with the real data given by SENASA, which provides latitude and longitude coordinates for the cited period. The results found for the movement of the cloud were satisfactory, they matched with the real data, identifying that temperature and wind speed have a great influence on the movement of locust swarms. The methodology used allows monitoring in real-time their movement, predicting the trajectory and making it possible to plan actions by government control agencies with pesticides in convenient areas.


Characterization of the iron-sulfur protein of the mitochondrial outer membrane partially purified from beef kidney cortex.
The iron-sulfur protein present in the mitochondrial outer membrane has been partially purified from beef kidney cortex mitochondria by means of selective solubilization followed by DEAE-cellulose chromatography. The EPR spectrum of the iron-sulfur protein with g-values at 2.01, 1.94 and 1.89 was well resolved up to 200 K which is unusual for an iron-sulfur protein. Analyses confirmed a center with two iron and two labile sulfur atoms in the protein. By measuring the effect of oxidation-reduction potential on the EPR signal amplitude, midpoint potentials at pH 7.2 were determined both for the purified iron-sulfur protein, +75 (+/- 5) mV, and in prepared mitochondrial outer membrane, +62 (+/- 6) mV. At pH 8.2 slightly lower values were indicated, +62 and 52 mV, respectively. The oxidation-reduction equilibrium involved a one electron transfer. A functional relationship to the rotenone-insensitive NADH-cytochrome c oxidoreductase in the mitochondrial outer membrane is suggested. Both this activity and the iron-sulfur center were sensitive to acidities slightly below pH 7 in contrast to the iron-sulfur centers of the inner membrane.


Three-dimensional microengineered vascularised endometrium-on-a-chip.
Can we reconstitute physiologically relevant 3-dimensional (3D) microengineered endometrium in-vitro model? Our representative microengineered vascularised endometrium on-a-chip closely recapitulates the endometrial microenvironment that consists of three distinct layers including epithelial cells, stromal fibroblasts and endothelial cells in a 3D extracellular matrix in a spatiotemporal manner. Organ-on-a-chip, a multi-channel 3D microfluidic cell culture system, is widely used to investigate physiologically relevant responses of organ systems. The device consists of five microchannels that are arrayed in parallel and partitioned by array of micropost. Two central channels are for 3D culture and morphogenesis of stromal fibroblast and endothelial cells. In addition, the outermost channel is for the culture of additional endometrial stromal fibroblasts that secrete biochemical cues to induce directional pro-angiogenic responses of endothelial cells. To seed endometrial epithelial cells, on Day 8, Ishikawa cells were introduced to one of the two medium channels to adhere on the gel surface. After that, the microengineered endometrium was cultured for an additional 5-6 days (total ∼ 14 days) for the purpose of each experiment. Microfluidic 3D cultures were maintained in endothelial growth Medium 2 with or without oestradiol and progesterone. Some cultures additionally received exogenous pro-angiogenic factors. For the three distinct layers of microengineered endometrium-on-a-chip, the epithelium, stroma and blood vessel characteristics and drug response of each distinct layer in the microfluidic model were assessed morphologically and biochemically. The quantitative measurement of endometrial drug delivery was evaluated by the permeability coefficients. We established microengineered vascularised endometrium-on-chip, which consists of three distinct layers: epithelium, stroma and blood vessels. Our endometrium model faithfully recapitulates in-vivo endometrial vasculo-angiogenesis and hormonal responses displaying key features of the proliferative and secretory phases of the menstrual cycle. Furthermore, the effect of the emergency contraception drug levonorgestrel was evaluated in our model demonstrating increased endometrial permeability and blood vessel regression in a dose-dependent manner. We finally provided a proof of concept of the multi-layered endometrium model for embryo implantation, which aids a better understanding of the molecular and cellular mechanisms underlying this process. N/A. This report is largely an in-vitro study and it would be beneficial to validate our findings using human primary endometrial cells. Our 3D microengineered vascularised endometrium-on-a-chip provides a new in-vitro approach to drug screening and drug discovery by mimicking the complicated behaviours of human endometrium. Thus, we suggest our model as a tool for addressing critical challenges and unsolved problems in female diseases, such as endometriosis, uterine cancer and female infertility, in a personalised manner. This work is supported by funding from the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIT) to Y.J.K. (No. 2018R1C1B6003), to J.A. (No. 2020R1I1A1A01074136) and to H.S.K. (No. 2020R1C1C100787212). The authors report no conflicts of interest.


Optical properties of an outer membrane lipoprotein from Escherichia coli.
The infrared spectrum of a structural lipoprotein from the Escherichia coli outer membrane indicated the lipoprotein had an alpha-helical conformation but no sign for the existence of beta-structures. From circular dichroism spectra of the lipoprotein, the alpha-helical content of the protein was found to be as high as 88% in 0.01-0.03% sodium dodecyl sulfate in the presence of 10(-5) M Mg2+ at pH 7.1 and 23 degrees C. When sodium dodecyl sulfate concentration increased higher than 0.1%, the alpha-helical content of the lipoprotein decreased to about 57%. Divalent cations, such as Mg2+ and Mn2+, were found to increase the helical content of the lipoprotein. The high alpha-helical content of the lipoprotein was observed in a wide range of temperatures (23 to 55 degrees C). The significance of the high alpha-helical content of the lipoprotein is discussed in light of the three-dimensional molecular models of the lipoprotein proposed previously.


A maxi Cl- channel coupled to endothelin B receptors in the basolateral membrane of guinea-pig parietal cells.
1. To study endothelin (ET) receptors in guinea-pig stomach, ET-binding assays and in vitro autoradiography were performed on fundic cell suspensions and on sections of the fundus, respectively. ETA and ETB receptor subtypes were found to coexist in the parietal cells. 2. Endothelin 1 (ET-1) added to the (basolateral) bathing solution was found to activate noisy whole-cell Cl- currents within about 1 min in both single, isolated parietal cells and those within gastric glands obtained from the fundus. 3. ET-1-induced Cl- currents were rapidly blocked by a Cl- channel blocker (NPPB) added to the (basolateral) bathing solution in a concentration-dependent manner with a half-maximum inhibition concentration of 33 microM. 4. The anion selectivity sequence of the ET-1-induced conductance was I- > Br- > Cl- > F-, corresponding to Eisenman's sequence I. 5. Changes in extracellular pH between 5 and 8 did not affect the ET-1-induced activation of Cl- currents. 6. Similar activating effects were also observed with ET-3 and a specific ETB receptor agonist (IRL1620). An ETB receptor antagonist (IRL1720) prevented the ET-1 effect, whereas an ETA-selective antagonist (FR139317 or BQ123) failed to antagonize the ET-1 effect. 7. In the whole-cell mode, unitary Cl- channel events could be observed in association with ET-1-activated macroscopic currents. The single-channel conductances were around 200 and 350 pS at negative and positive membrane potentials, respectively. 8. It is concluded that gastric parietal cells of guinea-pig possess pH-insensitive 'maxi' Cl- channels coupled to ETB receptors in the basolateral membrane.


Nanoadsorbents in focus for the remediation of environmentally-related contaminants with rising toxicity concerns.
Modern lifestyle demands high-end commodities, for instance, cosmetics, detergents, shampoos, household cleaning, sanitary items, medicines, and so forth. In recent years, these products' consumption has increased considerably, being antibiotics and some other pharmaceutical and personal care products (PPCPs). Several antibiotics and PPCPs represent a wide range of emerging contaminants with a straight ingress into aquatic systems, given their high persistence in seawater, effluent treatment plants, and even drinking water. Under these considerations, the necessity of developing new and affordable technologies for the treatment and sustainable mitigation of pollutants is highly requisite for a safer and cleaner environment. One possible mitigation solution is an effective deployment of nanotechnological cues as promising matrices that can contribute by attending issues and improving the current strategies to detect, prevent, and mitigate hazardous pollutants in water. Focused on nanoparticles' distinctive physical and chemical properties, such as high surface area, small size, and shape, metallic nanoparticles (MNPs) have been investigated for water remediation. MNPs gained increasing interest among research groups due to their superior efficiency, stability, and high catalyst activity compared with conventional systems. This review summarizes the occurrence of antibiotics and PPCPs and the application of MNPs as pollutant mitigators in the aquatic environment. The work also focuses on transportation fate, toxicity, and current regulations for environmental safety.


Factors influencing nominal effective concentrations of chemical compounds in vitro: medium protein concentration.
Quantitative data used to characterise biological activities of chemicals in vitro (e. g. EC50 values) are generally based on nominal concentrations and thus depend on factors influencing the availability of a compound. In this study, the impact of protein binding on the availability of chemicals in vitro is theoretically investigated and experimentally examined using a bovine sperm cell assay to measure the cytotoxic potency of selected compounds at different medium protein concentrations. In agreement with theoretical considerations, linear correlations between EC50 values and medium albumin concentrations were determined with 2,4-dichlorophenol, pentachlorophenol, p,p'-DDT and mercuric chloride. Ratios of EC50 values measured in the presence and absence of 4% (w/v) albumin varied between 500 (hexachlorophene), 258 (pentachlorophenol) and almost 1 (potassium cyanide, dextropropoxyphene). Calculated molar ratios of substance bound to albumin ranged from 0.05 (arsenic trioxide) and 0.1 (potassium cyanide) to 2.5 and 4.7 moles/mole for malathion and xylene, respectively. The fractions bound at 4% albumin varied between 11 and 15% for dextropropoxyphene and potassium cyanide, respectively, and more than 99% for hexachlorophene, pentachlorophenol and mercuric chloride. The results clearly demonstrate that the differing impact of protein binding on the bioavailability of chemicals considerably influences their nominal and relative potencies in the presence of albumin.


Surgical explantation of failed transcatheter heart valves: indications and results.
Given the recent surge in transcatheter heart valve replacement (THVR), cardiac surgeons will surely face the challenge of eventual explantation. The aim of this study was to determine indications for reoperation, while exploring pertinent technical aspects and survival after THV explantation in a cohort originally deemed high risk or even inoperable. Between February 2008 and March 2019, 31 patients with failed transcatheter aortic valve replacement (TAVR) underwent surgical explantations at our facility. Data were prospectively collected for retrospective analysis of procedural indications, technical issues, and postoperative survival. The major reason for TAVR removal was bioprosthetic valve failure (BVF) due to infective endocarditis (IE: 16/31 [51.6%]), non-structural (NSVD: 14/31 [45.2%]) and structural (SVD: 1/31 [3.2%]) valve deterioration accounting for the rest. Mean age at THV explantation was 76.3 ± 8.3 years, and median time from TAVR to explantation was 153 days (0 days-56.6 months). Median ICU and hospital stay were 6 days (1-44 days) and 23 days (8-62 days), respectively. Thirty-day and 1-year survival rates were 74.2% and 67.2%, respectively. Median follow-up interval after explantation was 364 days (3 days-80 months). Mean cardiopulmonary bypass time was 124.6 ± 46.8 min, and mean aortic cross-clamp time was 84.3 ± 32.9 min. There was no need for unplanned aortic root repair owing to tissue damage during dissection of the TAVR from surrounding tissue. The most common reason for THV explantation was (a) BVF for IE and (b) BVF secondary to NSVD. Although 30-day and 1-year mortality rates in this multimorbid cohort were predictably high, no procedural mortalities occurred.


Surface Markers Guide the Journey toward Naive Pluripotency.
Several protocols have managed to reset human primed PSCs to the naive state. In this issue of Cell Stem Cell, Collier et al. (2017) report a set of surface markers that identify which cells are susceptible to resetting and suggest a potential roadmap for the acquisition of naive pluripotency.


High activity of inclusion bodies formed in Escherichia coli overproducing Clostridium thermocellum endoglucanase D.
The formation of cytoplasmic inclusion bodies by Escherichia coli overproducing Clostridium thermocellum endoglucanase D (EGD) was investigated. EGD was found in inclusion bodies as a 68 kDa form, whereas the size of the cytoplasmic form was 65 kDa. Upon solubilization with urea followed by dialysis, the 68 kDa form was converted to the 65 kDa species. Proteolysis occurred within the COOH-terminal, reiterated region of the 68 kDa form, which is conserved among most C. thermocellum endoglucanases, but is not required for catalytic activity. The specific activity of the enzyme embedded in inclusion bodies was close to that of the purified protein. Thus, inclusion body formation does not involve denaturation of the catalytic domain of EGD, but, more likely, the participation of the reiterated, conserved region in intermolecular interactions.


Bi-directional flow in coronary-to-left ventricular fistula.
Coronary artery fistulae are rare congenital anomalies occasionally observed during coronary angiography. We present a case of retrograde coronary opacification during left ventriculography. This is the first reported case of angiographically documented bi-directional flow in these structures. Due to advanced age and personal wishes, the patient was treated medically. The epidemiology, pathophysiology, and therapy for coronary fistulae are briefly discussed.


Reversion of early- and late-stage β-cell dedifferentiation by human umbilical cord-derived mesenchymal stem cells in type 2 diabetic mice.
The authors aimed to observe β-cell dedifferentiation in type 2 diabetes mellitus (T2DM) and investigate the reversal effect of umbilical cord-derived mesenchymal stem cells (UC-MSCs) on early- and late-stage β-cell dedifferentiation. In high-fat diet (HFD)/streptozotocin (STZ)-induced T2DM mice, the authors examined the predominant role of β-cell dedifferentiation over apoptosis in the development of T2DM and observed the reversion of β-cell dedifferentiation by UC-MSCs. Next, the authors used db/db mice to observe the progress of β-cell dedifferentiation from early to late stage, after which UC-MSC infusions of the same amount were performed in the early and late stages of dedifferentiation. Improvement in metabolic indices and restoration of β-cell dedifferentiation markers were examined. In HFD/STZ-induced T2DM mice, the proportion of β-cell dedifferentiation was much greater than that of apoptosis, demonstrating that β-cell dedifferentiation was the predominant contributor to T2DM. UC-MSC infusions significantly improved glucose homeostasis and reversed β-cell dedifferentiation. In db/db mice, UC-MSC infusions in the early stage significantly improved glucose homeostasis and reversed β-cell dedifferentiation. In the late stage, UC-MSC infusions mildly improved glucose homeostasis and partially reversed β-cell dedifferentiation. Combining with other studies, the authors found that the reversal effect of UC-MSCs on β-cell dedifferentiation relied on the simultaneous relief of glucose and lipid metabolic disorders. UC-MSC therapy is a promising strategy for reversing β-cell dedifferentiation in T2DM, and the reversal effect is greater in the early stage than in the late stage of β-cell dedifferentiation.


Effects of hypobaria on parabronchial gas exchange in normoxic and hypoxic ducks.
Cardio-respiratory parameters and air sac and blood gases were measured in the unrestrained, unanesthetized duck during exposure to normobaric (PB = 746 Torr) or hypobaric (PB = 253 Torr) normoxia (PIO2 = 143 Torr) and hypoxia (PIO2 = 41.5 Torr). Compared with normobaria at the same PIO2, hypobaria caused a statistically significant increase in ventilation during both normoxia and hypoxia, resulting in elevated PO2 and diminished PCO2 in the caudal thoracic and clavicular air sac, and in increased PaO2 and decreased PaCO2. Similarly, lactic acid production was elevated in hypobaria, and the resulting decrease in arterial pH may be responsible for the increase in ventilation. Despite these changes, there was no evidence for altered gas exchange efficiency during hypobaria. This suggests that no significant diffusion limitation is present in the air capillary gas phase in normobaria, that could have been diminished with hypobaria. It also suggests that the aerodynamic valving efficiency, present during inspiration at the level of the medioventral bronchi, is not affected by hypobaria. Although the mechanisms underlying the increased lactic acid production and ventilation are not understood, they may exert an advantageous effect on high altitude tolerance of the bird.


Corrigendum: Image discrimination reversal learning is impaired by sleep deprivation in rats: Cognitive rigidity or fatigue?
[This corrects the article DOI: 10.3389/fnsys.2022.1052441.].


Multi-element contamination in soils from major mining areas in Northeastern of Brazil.
Mining has become one of the main factors in the global biogeochemical cycle of potentially toxic elements. Therefore, it is considered one of the anthropogenic activities with the greatest negative impact on the environment. These impacts are maximized in semiarid regions, where mining activities can lead to soil degradation and decrease in land productivity. This study aimed to assess the level of contamination in natural, urban, and agricultural soils of three important mining areas, where approximately 80,000 people live, and pollution levels have never been determined before. For this purpose, soil samples were collected around iron, uranium, and vanadium mines, as well as in the main human settlements of the region. The concentrations of 34 elements were determined by instrumental neutron analysis activation (INAA) and inductively coupled plasma optical emission spectrometry (ICP OES) techniques. Pollution indices (CF, EF, mCd, PLI, and REEP) revealed that there is a moderate to heavy level of pollution for 89% of the analyzed elements. Additionally, an extreme contamination level was observed in 78% of the samples, for at least one element. Statistical analyses were performed to identify patterns in the distribution and common sources of pollution. The results suggest that the concentrations for Al, Ba, Hf, Na, Pb, Rb, REE, Ta, Th, U, Zn, and Zr are associated with geogenic causes. However, the influence of anthropogenic sources such as agriculture and mining on the accumulation of these elements in soils should not be disregarded. In contrast, the contents of As, Br, Cd, Co, Cr, Cs, Cu, Fe, K, Mn, Ni, Sc, Ti, and V reflect the direct impact of anthropogenic sources.


Disentangling the energetic costs of step time asymmetry and step length asymmetry in human walking.
The metabolic cost of walking in healthy individuals increases with spatiotemporal gait asymmetries. Pathological gait, such as post-stroke, often has asymmetry in step length and step time which may contribute to an increased energy cost. But paradoxically, enforcing step length symmetry does not reduce metabolic cost of post-stroke walking. The isolated and interacting costs of asymmetry in step time and step length remain unclear, because previous studies did not simultaneously enforce spatial and temporal gait asymmetries. Here, we delineate the isolated costs of asymmetry in step time and step length in healthy human walking. We first show that the cost of step length asymmetry is predicted by the cost of taking two non-preferred step lengths (one short and one long), but that step time asymmetry adds an extra cost beyond the cost of non-preferred step times. The metabolic power of step time asymmetry is about 2.5 times greater than the cost of step length asymmetry. Furthermore, the costs are not additive when walking with asymmetric step time and asymmetric step length: the metabolic power of concurrent asymmetry in step length and step time is driven by the cost of step time asymmetry alone. The metabolic power of asymmetry is explained by positive mechanical power produced during single support phases to compensate for a net loss of center of mass power incurred during double support phases. These data may explain why metabolic cost remains invariant to step length asymmetry in post-stroke walking and suggest how effects of asymmetry on energy cost can be attenuated.


MARC: A Novel Framework for Detecting MITM Attacks in eHealthcare BLE Systems.
Real-time and ubiquitous patient monitoring demands the use of wireless data acquisition through resource constrained medical sensors, mostly configured with No-input No-output (NiNo) capabilities. Bluetooth is one of the most popular and widely adopted means of communicating this sensed information to a mobile terminal. However, over simplified implementations of Bluetooth low energy (BLE) protocol in eHealth sector is susceptible to several wireless attacks, in particular the Man-in-the-Middle (MITM) attack. The issue arises due to a lack of mutual authentication and integrity protection between the communicating devices, which may lead to compromise of confidentiality, availability and even the integrity of this safety-critical information. This research paper presents a novel framework named MARC to detect, analyze, and mitigate Bluetooth security flaws while focusing upon MITM attack against NiNo devices. For this purpose, a comprehensive solution has been proposed, which can detect MITM signatures based upon four novel anomaly detection metrics: analyzing Malicious scan requests, Advertisement intervals, RSSI levels, and Cloned node addresses. The proposed solution has been evaluated through practical implementation and demonstration of different attack scenarios, which show promising results concerning accurate and efficient detection of MITM attacks.


Serum potassium and uric acid changes during treatment with timolol alone and in combination with a diuretic.
Timolol, 10 to 40 mg daily, given to 103 patients with uncomplicated arterial hypertension induced significant increments of serum potassium at all dose levels (p less than 0.05). The magnitude of the increments was dependent on daily timolol dosage. When hydrochlorothiazide and amiloride were added, serum potassium decreased (p less than 0.001), but a major determinant of the magnitude of the decrease was the dosage change of the timolol. Serum uric acid was influenced in a paradoxical way during timolol monotherapy; there was a rise in all 3 dosage groups (p less than 0.02) but the lowest group showed the largest increase and vice versa. On addition of hydrochlorothiazide and amiloride, there was a further increase in serum uric acid, the magnitude of which depended on the concomitant reduction in the dose of timolol, with reductions in dose causing a larger rise in serum uric acid and increments, a smaller rise. The increments of serum uric acid were greater in females than in males during both treatment periods. The results indicate that beta blockers induce dose-dependent rises in serum potassium and may counteract undesirable effects of diuretics on serum potassium. Beta blockers seem to have a paradoxical effect on serum uric acid and may aggravate the hyperuricemia induced by diuretics.


Decreased lipopolysaccharide content and enhanced susceptibility of leptospiras to serum leptospiricidal action and phagocytosis after treatment with diphenylamine.
Growth of leptospiras in the presence of diphenylamine (DPA) caused a decrease in the content of leptospiral lipopolysaccharide (LPS). In association with this decrease of LPS, leptospiras became susceptible to anti-leptospiral action of normal rabbit serum (NRS), leptospiricidal action of antibody and complement, and killing by phagocytes. DPA-treated leptospiras were eliminated rapidly from the blood of infected mice and could not grow in the animals.


Accuracy of RGB-D camera-based and stereophotogrammetric facial scanners: a comparative study.
This study aimed to evaluate and compare the accuracy and inter-operator reliability of a low-cost red-green-blue-depth (RGB-D) camera-based facial scanner (Bellus3D Arc7) with a stereophotogrammetry facial scanner (3dMD) and to explore the possibility of the former as a clinical substitute for the latter. A mannequin head was selected as the research object. In the RGB-D camera-based facial scanner group, the head was continuously scanned five times using an RGB-D camera-based facial scanner (Bellus3D Arc7), and the outcome data of each scan was then imported into CAD software (MeshLab) to reconstruct three-dimensional (3D) facial photographs. In the stereophotogrammetry facial scanner group, the mannequin head was scanned with a stereophotogrammetry facial scanner (3dMD). Selected parameters were directly measured on the reconstructed 3D virtual faces using a CAD software. The same parameters were then measured directly on the mannequin head using the direct anthropometry (DA) method as the gold standard for later comparison. The accuracy of the facial scanners was evaluated in terms of trueness and precision. Trueness was evaluated by comparing the measurement results of the two groups with each other and with that of DA using equivalence tests and average absolute deviations, while precision and inter-operator reliability were assessed using the intraclass correlation coefficient (ICC). A 3D facial mesh deviation between the two groups was also calculated for further reference using a 3D metrology software (GOM inspect pro). In terms of trueness, the average absolute deviations between RGB-D camera-based and stereophotogrammetry facial scanners, between RGB-D camera-based facial scanner and DA, and between stereophotogrammetry facial scanner and DA were statistically equivalent at 0.50±0.27 mm, 0.61±0.42 mm, and 0.28±0.14 mm, respectively. Equivalence test results confirmed that their equivalence was within clinical requirements (<1 mm). The ICC for each parameter was approximately 0.999 in terms of precision and inter-operator reliability. A 3D facial mesh analysis suggested that the deviation between the two groups was 0.37±0.01 mm. For facial scanners, an accuracy of <1 mm is commonly considered clinically acceptable. Both the RGB-D camera-based and stereophotogrammetry facial scanners in this study showed acceptable trueness, high precision, and inter-operator reliability. A low-cost RGB-D camera-based facial scanner could be an eligible clinical substitute for traditional stereophotogrammetry. The low-cost RGB-D camera-based facial scanner showed clinically acceptable trueness, high precision, and inter-operator reliability; thus, it could be an eligible clinical substitute for traditional stereophotogrammetry.


Acoustical dead zones and the spatial aggregation of whale strandings.
Cetacean strandings display a marked geographical clustering. We propose a simple, two-dimensional ray-dynamics model of cetacean echolocation to examine the role played by coastline topography in influencing the location and clustering of stranding sites. We find that a number of coastlines known to attract cetacean strandings produce acoustical "Dead Zones" where echolocation signals are severely distorted by purely geometric effects. Using available cetacean stranding data bases from four disparate areas, we show that the geographical clusters in the observations correlate strongly with the regions of distorted echolocation signals as predicted by the model.


Plasma interleukin-10 levels and adverse outcomes in acute coronary syndrome.
PURPOSE OR BACKGROUND: Interleukin (IL)-10 is an immunoregulatory cytokine that is produced by a variety of cell types, such as macrophages and activated monocytes. IL-10 possesses numerous anti-inflammatory, anti-thrombotic and anti-atherosclerotic properties. Furthermore, patients with acute coronary syndrome have been demonstrated to have reduced levels of IL-10 compared to their stable counterparts. For these reasons, it has been proposed that IL-10 plays a protective role in both atherogenesis and plaque vulnerability. However, 2 short-term studies on the prognostic utility of IL-10 in patients with acute coronary syndrome have provided conflicting results, with one study showing that reduced levels of IL-10 were predictors of adverse outcomes and another showing that elevated levels predicted poor outcomes. The objective of the present study was to investigate the long-term prognostic significance of baseline IL-10 levels in patients with acute coronary syndrome. Baseline plasma IL-10 levels were measured in 193 well-characterized male patients with acute coronary syndrome who were referred for coronary angiography and followed prospectively for 5 years for the development of major adverse cardiovascular events. After controlling for a variety of baseline variables (including established biomarkers such as high-sensitivity C-reactive protein and N-terminal-pro-B-type natriuretic peptide), plasma IL-10 levels (whether analyzed as a continuous variable or as a categorical variable using receiver operating characteristic-derived cut point) were a strong and independent predictor of the composite outcome of death or non-fatal myocardial infarction when using a Cox proportional hazards model. These data demonstrate that, despite biologic plausibility for IL-10 as being a cardioprotective cytokine, elevated baseline plasma levels of IL-10 are a strong and independent predictor of long-term adverse cardiovascular outcomes in patients with acute coronary syndrome.


Recognition highlights: Toll-like receptors.
As part of our first line of defense against invading organisms, Toll-like receptors recognize bacterial and viral molecules, and launch an inflammatory response.


Enhanced wound-healing efficacy of electrospun mesoporous hydroxyapatite nanoparticle-loaded chitosan nanofiber developed using pluronic F127.
One of the goals of wound repairing is to mimic the function and architecture of the native extracellular matrix (ECM). To this end, for the first time, we used pluronic F127 and mesoporous rod-like hydroxyapatite nanoparticles (mr-HAP NPs) simultaneously to prepare a novel low-diameter electrospun ECM-mimicking wound dressing based on a mixture of chitosan and polyethylene oxide. F127 is used as a surface tension regulator of the polymer solution. In addition, F127 has the special ability to reduce the size of nanofibers. mr-HAP NPs are used as cell proliferation accelerators which also improve the mechanical properties and water uptake capacity of the as-prepared dressing. The average size of nanofibers in the presence of F127 was about 110 nm which was more than 2.5 times lower than nanofibers prepared without F127. The water uptake capacity was evaluated to investigate the wound exudate absorption capacity of the wound dressing. It was observed that the incorporation of mr-HAP NPs into wound dressing structure increases the water uptake capacity by more than 2.5 times. Alongside the evaluation of cytocompatibility through in vitro cell viability assay, the wound healing efficacy was also determined in full-thickness skin wounds in a rat model for 15 days. The cytocompatible wound dressing showed significantly higher wound closure efficacy than the control group so the wounds healed entirely on the last day of the treatment period. As well, the pathology analysis proved better granulation tissue development and greater re-epithelialization. These findings are by virtue of the improved mechanical properties, accelerated cell migration and proliferation, proper environment for oxygen exchange, and enhanced exudate uptake of the wound dressing. These all are due to the presence of F127 and mr-HAP.


Use it or lose it? Lessons learned from the developing brains of children who are deaf and use cochlear implants to hear.
In the present paper, we review what is currently known about the effects of deafness on the developing human auditory system and ask: Without use, does the immature auditory system lose the ability to normally function and mature? Any change to the structure or function of the auditory pathways resulting from a lack of activity will have important implications for future use through an auditory prosthesis such as a cochlear implant. Data to date show that deafness in children arrests and disrupts normal auditory development. Multiple changes to the auditory pathways occur during the period of deafness with the extent and type of change being dependent upon the age and stage of auditory development at onset of deafness, the cause or type of deafness, and the length of time the immature auditory pathways are left without significant input. Structural changes to the auditory nerve, brainstem, and cortex have been described in animal models of deafness as well in humans who are deaf. Functional changes in deaf auditory pathways have been evaluated by using a cochlear implant to stimulate the auditory nerve with electrical pulses. Studies of electrically evoked activity in the immature deaf auditory system have demonstrated that auditory brainstem development is arrested and that thalamo-cortical areas are vulnerable to being taken over by other competitive inputs (cross-modal plasticity). Indeed, enhanced peripheral sight and detection of visual movement in congenitally deaf cats and adults have been linked to activity in specific areas of what would normally be auditory cortex. Cochlear implants can stimulate developmental plasticity in the auditory brainstem even after many years of deafness in childhood but changes in the auditory cortex are limited, at least in part, by the degree of reorganization which occurred during the period of deafness. Consequently, we must identify hearing loss rapidly (i.e., at birth for congenital deficits) and provide cochlear implants to appropriate candidates as soon as possible. Doing so has facilitated auditory development in the thalamo-cortex and allowed children who are deaf to perceive and use spoken language.


Breast density changes associated with hormone replacement therapy in postmenopausal women. Effects on the specificity and sensitivity of mammographic screening.
Postmenopausal HRT use is associated with an increase of mammographic density and reduction of sensitivity and specificity of mammography results and an increase of false-positive and false-negative outcomes. The increased density does not allow a good evaluation of the exam. Mammographic density is an independent risk factor for breast cancer, but the link between changes in breast density and difference in breast cancer risk, remain uncertain. On the other hand, today specific guidelines and protocols to optimize the screening of neoplastic breast pathology in HRT users do not exist and it is unknown if short-term suspension of therapy improves mammographic sensitivity. More information is required to define this important risk factor.


Mitral valve stenosis caused by abnormal pannus extension over the prosthetic ring and leaflets after Duran ring mitral annuloplasty.
We treated a 61-year-old woman with mitral stenosis caused by pannus formation after Duran ring annuloplasty. Pannus overgrowth on the ring with extension onto both leaflets narrowed the mitral orifice and severely restricted the mobility of the valve leaflets. Mitral valve replacement with a St. Jude Medical mechanical heart valve prosthesis was successfully performed, and the postoperative course was uneventful. Patients undergoing Duran ring annuloplasty should be followed up with the consideration of possible mitral stenosis caused by pannus extension, as the cause for pannus formation remains unclear.


[The effect of NaCl solution with different concentration on the resuscitation of hemorrhagic shock].
To observe the clinical results of different concentration NaCl solution in hemorrhagic shock resuscitation. The 75 cases with hemorrhagic shock were divided into five groups by means of random number table. Treat the cases with 0.9% (control group), 3%, 4.5%, 6% and 7.5% NaCl solution, respectively. Observe the changes of BP, HR, blood gas, renal function, electrolyte concentration of plasma, before and after treatment. After transfusion, compared with groups of 0.9% NS, the mean BP of patients in other four groups rise more quickly but only the group of 4.5% can last 30 minutes (P < 0.05), and K(+) of plasma in groups of 6%, 7.5% NS decline more obviously (P < 0.05), the group of 4.5% maintain the best lever. The mean BP of patients rise quickly and continue for a long time, in the group of 4.5%. Meanwhile the internal environment of the body maintain relatively stable in the group of 4.5%.


Sleep-dependent motor memory consolidation in healthy adults: A meta-analysis.
It is widely accepted that sleep better facilitates the consolidation of motor memories than does a corresponding wake interval (King et al., 2017). However, no in-depth analysis of the various motor tasks and their relative sleep gain has been conducted so far. Therefore, the present meta-analysis considered 48 studies with a total of 53 sleep (n = 829) and 53 wake (n = 825) groups. An overall comparison between all sleep and wake groups resulted in a small effect for the relative sleep gain in motor memory consolidation (g = 0.43). While no subgroup differences were identified for differing designs, a small effect for the finger tapping task (g = 0.47) and a medium effect for the mirror tracing task (g = 0.62) were found. In summary, the meta-analysis substantiates that sleep generally benefits the consolidation of motor memories. However, to further our understanding of the mechanisms underlying this effect, examining certain task dimensions and their relative sleep gain would be a promising direction for future research.


Post COVID-19 pneumo-hematocele: clinical presentation and treatment pathway.
The appearance of characteristic pulmonary lesions has been noted after COVID-19, being described as post-COVID-19 pneumo-hematocele. The aim of this study is to describe the clinical, histopathologic, and imaging features of pneumo-hematocele and to suggest a treatment algorithm for these patients. A retrospective study was performed in patients admitted with a diagnosis of SARS-CoV2 infection from March 2020 to September 2021 who presented a pneumo-hematocele on imaging studies. Clinical and demographic variables were recorded, and CT scans were analyzed. A secondary analysis was performed to estimate the risk provided by the pneumo-hematocele diameter of developing pneumothorax. 37 patients were diagnosed with pneumo-hematoceles, 97.3% were males with a median age of 41 ± 13 years and 51% were smokers. The mean diameter of the pneumatocele was 6.3 ± 2.8 cm; they were more common on the subpleural surface and in the inferior lobe. Thirty patients had ruptured pneumo-hematoceles and developed pneumothorax (81.1%); a total of 26 patients required surgery (70.3%). Lesions measuring 5 cm had a high risk of rupture (OR 6.8, CI 95% 1.1-42); those measuring 3 cm were prone to this complication. For each centimeter that the pneumo-hematocele diameter increases, the OR for rupture increases 1.5. It appears that post-COVID-19 pneumo-hematocele occurs secondary to encapsulation of blood accumulation inside the lung, as a result of micro-capillary bleeding, with partial reabsorption of blood and subsequent air filling. We recommend surgery for patients with pneumo-hematoceles of 5 cm and those with persistent lesions of 3 cm. Clinical Trial Registration: NCT05067881.


Synthesis and biological evaluation of benzyl styrylsulfonyl derivatives as potent anticancer mitotic inhibitors.
We herein report the synthesis, biological activity and structure activity relationship of derivatives of benzylstyrylsulfone, benzylstyrylsulfine and benzylsulfonyl-N-phenylacetamide. A lead compound 7 represents a new class of mitotic inhibitors that demonstrates potent anti-proliferative activity and selectively induces cancer cell apoptosis while sparing non-transformed lung fibroblast.


Truncated, Non-networking Versions of the Coupling Protein CheW Retain Chemoreceptor Control of Kinase CheA.
Bacterial chemoreceptors control the activity of the associated CheA kinase in response to chemical gradients and, consequently, regulate the swimming behavior of the cell. However, such control is not direct but requires the participation of the essential coupling protein CheW, which is structurally homologous to the carboxy-terminal domain of the kinase. The actual role of this small coupling protein is somehow intriguing. It has been demonstrated that it is absolutely essential for chemoreceptor control of the kinase, in spite of the occurrence of direct contacts between chemoreceptors and CheA. In addition, CheW plays an essential role in the assembly of the large macromolecular arrays that combine chemoreceptors of different specificities, and it is therefore responsible for molecular interactions that provide such arrays with remarkable signaling properties. In this work, we analyze truncated CheW derivatives that are still able to control the kinase but have lost the ability to connect signaling units. We demonstrate that these two activities can work separately and speculate about the significance of the roles of these two different activities in the context of the chemoreceptor cluster.


Adherence to a Mediterranean diet, dyslipidemia and inflammation in familial hypercholesterolemia.
Familial Hypercholesterolemia (FH) is characterized by elevated LDL-cholesterol (LDL-C) and high atherosclerosis risk. The impact of different dietary patterns on atherosclerosis biomarkers has been poorly studied in FH. This study verified the association of adherence to a Mediterranean diet with biomarkers of dyslipidemia and low-grade inflammation in molecularly proven FH adults from Brazil (BR) and Spain (SP). In this cross-sectional study adherence to the Mediterranean diet was assessed by a validated score and generalized estimating equations were used to evaluate its association with plasma LDL-C, apolipoprotein-B (ApoB) and high sensitivity C-reactive protein (hs-CRP) concentrations. We included 92 (mean age 45 years, 58.7% females) and 98 FH individuals (mean age 46.8 years, 60.2% females) respectively from BR and SP. FH causing variants did not differ between countries. LDL-C, ApoB and hs-CRP concentrations were higher in BR than in SP: 179 (135-250) and 161 (133-193) mg/dL; 141 (109-181) and 103 (88-134) mg/dL; and 1.6 (0.8-4.0) and 0.8 (0.4-1.5) mg/L respectively (all p < 0.001). Most of BR had low adherence (n = 77, 83.7%), while the majority of SP were divided into moderate (n = 35, 35.7%) and strong adherence to the Mediterranean diet (n = 37, 37.8%), p < 0.001. There was a significant inverse association of adherence to the Mediterranean diet score with higher LDL-C, ApoB, and hs-CRP after adjusting for socio economic parameters, caloric and fatty acid intakes as well as pharmacological lipid lowering therapies. Higher adherence to a Mediterranean diet was associated with better dyslipidemia and low-grade inflammation profiles in FH.


Editorial on the Research Topic the 2nd Edition of Mountain Sports Activities: Injuries and Prevention.
Mountain sports are continuously gaining popularity, currently fueled by the post-pandemic period expanding travel opportunities and the desire to escape the increasingly hot environmental conditions of urban areas-ambient temperature decreases by about 6 [...].


A randomized, double-blind, controlled trial of testosterone gel treatment versus vehicle control on the facial hair of young men with beta-thalassemia major.
To determine whether the medical prescription of testosterone gel can successfully increase facial hairs in young men with beta-thalassemia major. Thirty-two men with beta-thalassemia major, at least 15 years old (tanner stage 4), were randomized in four blocks according to age and serum testosterone level. The cases received 2.5% testosterone gel. The changes in the number of terminal hairs were evaluated by a dermatologist after 6 months. Student's t-test and paired t-test were used to compare the results. The serum testosterone levels of controls and cases were 9.5+/-5.7 (mean+/-SD) and 10.5+/-9.6 ng/l, respectively. The number of terminal hairs (per cm2) in cases (29.8+/-13.6) was significantly higher than that for controls (13.9+/-13.2) (p<0.005). One patient from each group complained of a sense of irritation. The 2.5% testosterone gel was effective and well tolerated in inducing the transformation of the terminal hairs of the beard area of thalassemic males.


The effect of Baroque music on behavioural disturbances in patients with dementia.
To study the effect of Baroque music in people with dementia. Patients in a multicultural dementia-specific aged care facility were subjected to Baroque music in a cross-over study. There were significantly more behavioural disturbances during the weeks when Baroque music was played compared to control periods (0.2 more episodes per week, P = 0.01), and more episodes in the afternoon shift compared to evening shift (1 more episode per week, P = 0.001). These results confirm that music can influence behaviour but is contrary to previous results.


Geographic maldistribution of general surgery PGYI residents: another US surgical desert.
Practicing general surgeons are unevenly distributed across the country. This study evaluates the geographic distribution of categorical, general surgery (GS) PGYI positions per capita. Data were obtained from the 2012 National Resident Matching Program match and the 2010 US Census. The mean for GS PGYI positions per 10(6) population was 3.85 ± .61; 27 states fell below this value. The 7 American College of Surgeons (ACS) regions ranged from a low of 1.4 ± .50 (Intermountain) to a high of 9.89 ± 4.41 (Northeast). The mean (2.18 ± .34) for the 19 state membership of the Southwestern Surgical Congress was below the mean for the country. There is a maldistribution of GS PGYI positions compared with state and regional populations, particularly in rural areas. This mirrors the maldistribution of practicing general surgeons across the United States. Additional GS residences and resident positions are urgently needed to correct this "Surgical Desert" of graduate surgical education.


One-year follow-up of a new posterior chamber toric phakic intraocular lens implantation for moderate-to-high myopic astigmatism.
To investigate refractive and visual outcomes as well as rotational stability following implantation of Eyecryl phakic toric intraocular lens (pIOL) for moderate-to-high myopic astigmatism. The efficacy, safety, predictability, stability, and adverse events of Eyecryl toric pIOL were evaluated in patients with spherical refraction from - 4.50 to - 18.50 diopters (D) and cylindrical refraction from - 0.50 to - 5.50 D. This study included 60 eyes of 31 patients. The mean manifest refraction spherical equivalent (MRSE) dropped from - 10.45 ± 2.74 D preoperatively to - 0.34 ± 0.51 D and - 0.40 ± 0.56 D at 6 and 12 months postoperatively, respectively. There was an 81% decrease in astigmatism after surgery. The safety and efficacy of indices were 1.36 ± 0.43 and 1.20 ± 0 .32. At the final follow-up, the rate of eyes within ± 1.00 D and ± 0.50 D of the desired MRSE were 85% and 68.33%, respectively. Vision-threatening complications were not observed during the follow-up. The implantation of pIOL was effective, safe, and predictable in patients with moderate-to-high myopic astigmatism during 1-year follow-up.


Charge-transfer complexes of pyrimidine Schiff bases with aromatic nitro compounds.
Charge-transfer (CT) complexes of pyrimidine Schiff bases, derived from condensation of 2-aminopyrimidine and substituted benzaldehydes, with some aromatic polynitro compounds were prepared and investigated using IR, UV, visible and (1)H NMR spectroscopy. For all solid complexes, the main interaction between the donor and acceptor molecules takes place through the π-π* interaction. Strong and some weak acidic acceptors, in addition interact through proton transfer from the acceptor molecule to the basic centre of the electron donor. Also, an n-π* transition was detected in some complexes.


Expression of stemness markers in mouse parthenogenetic-diploid blastocysts is influenced by slight variation of activation protocol adopted.
The importance of obtaining stem cells through alternative methods has increased progressively in the recent years due to the potential role that embryonic stem (ES) cells play in the field of regenerative medicine. In this regard, generation of parthenogenetic blastocysts allows the production of ethic-free ES cells without the need to manipulate normal embryos. Our work was aimed at clarifying whether variations in the method adopted to generate diploid parthenogenetic blastocysts could determine differences in the quality of blastocysts produced. In vitro development of mouse oocytes activated with three protocols, using Sr2+ and cytochalasin for different time, was compared with that of in vivo fertilized embryos. We have evaluated the efficiency of blastocyst formation and analysed the expression pattern of the stemness markers OCT4, CDX2, and NANOG. Our results indicate that the yield of diploid parthenogenotes and the segregation of the stemness marker OCT4 in the developing blastocyst are influenced by the parthenogenetic protocol adopted. Particularly, even if all methods tested allowed the production of blastocysts in vitro, the correct segregation of OCT4 occurred only in blastocysts developed from oocytes concomitantly treated for 4 h with Sr2+ and cytochalasin D. Our results indicate that the protocol employed to develop parthenogenetic blastocysts in vitro affects the quality of cells in the inner cell mass.


Unveiling the structural and energetic properties of thiazole-water complex by microwave spectroscopy and theoretical calculations.
The structure and non-covalent bonding features of the complex of thiazole and water were studied by using supersonic jet Fourier transform microwave spectroscopy and theoretical calculations. One isomer was observed which corresponds to the global minimum of the complex predicted theoretically. The rotational spectra of 9 additional isotopologues, including 5 mono-substituted heavy atoms of thiazole (34S, 13C and 15N), and 4 water isotopic species (H218O, DOH, HOD and D2O), were also measured and analyzed. The experimental spectroscopic parameters were used to determine the structural parameters of the observed isomer. Theoretical analyses based on quantum theory of atoms in molecules and natural bond orbital revealed that the two moieties are linked by a N···H-O hydrogen bond. The total interaction energy of the complex is calculated to be -25.1 kJmol-1 with electrostatics being the major term according to energy decomposition analysis.


Left atrial dissection after a supra-annular mitral valve replacement for endocarditis.
Left atrial dissection is a rare and a potentially fatal complication of cardiac surgery. Multi-modal imagery is helpful for the diagnosis and to guide the treatment. We report the case of a 66-year-old female patient who underwent a combined mitral and aortic valve replacement for degenerative valvular disease. She presented an infectious endocarditis revealed by a third-degree atrioventricular bloc and had a redo mitral- and aortic valve replacement. Mitral valve was inserted in supra-annular position due to annular destruction. Post-operative course was marked by a refractory acute heart failure explained by a left atrial wall dissection confirmed by transoesophageal echocardiography and synchronized cardiac CT-scan. Surgical treatment was theoretically indicated but considering the high risk of a third surgery, a palliative care support was collegially decided. Left atrial dissection can occur after a redo surgery and supra-annular mitral valve implantation. Multi-modal imagery including transoesophageal echocardiography and cardiac CT-scan is helpful for the diagnosis.


Closing the loopholes on a flavored cigar ban: Anticipated challenges and solutions.
For the first time since Congress authorized the U.S. Food and Drug Administration (FDA) to regulate tobacco in 2009, FDA proposed two tobacco product standards on May 4, 2022. After a period of public comment and agency review, if the product standards are finalized in their current form, they will ban menthol cigarettes (U.S. Food and Drug Administration, 2022b), which comprised 37% of the U.S. cigarette market in 2019 and 2020 (Federal Trade Commission, 2021), and flavored cigars (U.S. Food and Drug Administration, 2022a), which have exceeded 50% of cigar sales since 2012 (Delnevo et al., 2021b). This commentary examines potential challenges to banning flavored cigars and highlights issues FDA and Congress should consider.


The Control Centers of Biomolecular Phase Separation: How Membrane Surfaces, PTMs, and Active Processes Regulate Condensation.
Biomolecular condensation is emerging as an essential process for cellular compartmentalization. The formation of biomolecular condensates can be driven by liquid-liquid phase separation, which arises from weak, multivalent interactions among proteins and nucleic acids. A substantial body of recent work has revealed that diverse cellular processes rely on biomolecular condensation and that aberrant phase separation may cause disease. Many proteins display an intrinsic propensity to undergo phase separation. However, the mechanisms by which cells regulate phase separation to build functional condensates at the appropriate time and location are only beginning to be understood. Here, we review three key cellular mechanisms that enable the control of biomolecular phase separation: membrane surfaces, post-translational modifications, and active processes. We discuss how these mechanisms may function in concert to provide robust control over biomolecular condensates and suggest new research avenues that will elucidate how cells build and maintain these key centers of cellular compartmentalization.


ADAM7 promotes the proliferation and invasion in trophoblast cells.
ADAMs are members of the metzincin family of zinc-dependent metalloproteinases, which play a key role in the proteolytic degradation of the extracellular matrix to invade cells. It is well known that ADAMs are involved in regulating the invasion of trophoblast cells. But the function and underlying mechanism of ADAM7 in trophoblast cells is still unknown. ADAM7 knockdown strongly inhibited HTR-8 and B6Tert-1 cells proliferation, migration and invasion, while ADAM7 overexpression reversed. The expression of protein pro-caspase 3 and pro-caspase 9 was not affected by either ADAM7 knockdown or overexpression in HTR-8 and B6Tert-1 cells, while the expression of active-caspase 3 and active-caspase 9 was strongly increased in ADAM7 silenced cells and significant decreased in ADAM7 overexpressed cells. We also found that the phosphorylation of p38MAPK was significantly inhibited in ADAM7 silenced cells while it was significantly induced in ADAM7 overexpressed cells. Metformin HCl could reverse the inhibitory effects of ADAM7 knockdown on the p38MAPK signaling pathway and the proliferation of HTR-8 and B6Tert-1 cells. ADAM7 plays a positive role in trophoblast cells, which may be attributed to regulation of the p38MAPK signaling pathway. SIGNIFICANCE: It is necessary to further study the molecular mechanism of trophoblast cells proliferation, migration and invasion, and develop a more effective treatment to preeclampsia. This research might provide a new target for further research in this area.


Evaluating relationships among clinical working memory assessment and inattentive and hyperactive/impulsive behaviors in a community sample of children.
This study examined relationships between inattentive and hyperactive/impulsive behaviors and working memory (WM) functioning, and the utility of WM in categorical diagnosis of ADHD versus considering ADHD symptoms on a continuum. The study included 50 male children (6-12 years). Inattentive and hyperactive/impulsive behaviors were measured by the Conners-3P parent report, and WM was assessed by the WISC-IV WM subtests and Working Memory Index (WMI). WISC-IV Arithmetic and Digit Span Backward were most consistently related to inattentive behaviors, and no WM measure was consistently related to ADHD hyperactive/impulsive behaviors. Arithmetic and Digit Span Backward also accounted for significant variance in inattentive behaviors and ADHD inattention symptoms, respectively. Neither the WMI nor the Arithmetic subtest correctly classified individuals diagnosed with ADHD. Measurement of inattentive behaviors on a continuum best characterized relationships between symptoms of ADHD and WM functioning; WM functioning did not have utility in categorical understanding of ADHD.


Purpurogallin improves neurological functions of cerebral ischemia and reperfusion mice by inhibiting endoplasmic reticulum stress and neuroinflammation.
Purpurogallin (PPG) has been testified to have neuroprotective effects. This study intends to probe the neuroprotection of PPG on cerebral ischemia/reperfusion (I/R) injury and its potential mechanism. C57/B6 mice, BV2 microglia and HT22 hippocampal neurons were used for in-vivo and in-vitro experiments. I/R injury models were constructed using middle cerebral artery occlusion (MCAO/R) and oxygen-glucose deprivation/reoxygenation (OGD/R), respectively. The expression of apoptosis and inflammatory proteins, and endoplasmic reticulum (ER) stress proteins were gauged by Western blotting (WB). The contents of inflammatory cytokines in OGD/R-induced BV2 microglia were testified by enzyme-linked immunosorbent assay (ELISA). Cell counting kit-8 (CCK-8), TUNEL assay and flow cytometry (FCM) were utilized to examine the viability and apoptosis of cells. The neurological, learning and memory functions were evaluated by the modified neurological severity score (mNSS) and water maze experiment. 2, 3, 5-triphenyltetrazole chloride (TTC) staining was utilized to calculate the volume of cerebral infarction and cerebral edema in the peri-infarct area. Apoptosis-related proteins, inflammation-related proteins and ER stress proteins were gauged by WB. ELISA was conducted to verify inflammatory cytokines. PPG treatment notably abated the expression of ER stress proteins and inflammatory factors in OGD/R-induced BV2 microglia and boosted HT22 neuron's viability and eased their apoptosis in comparison to the control group. In vivo, PPG treatment signally lessened cerebral infarct area, cerebral edema, and neurological deficit scores in MCAO/R mice. Additionally, PPG caused a dramatic decline in neuronal apoptosis and levels of ER stress proteins and inflammatory factors in the brain's peri-infarct region of MCAO/R mice. Mechanically, PPG blocked the TLR4/NF-κB pathway in OGD/R-induced BV2, HT22 neurons, and the MCAO/R mice. PPG attenuates brain I/R damage probably by suppressing ER stress and neuroinflammation via inactivation of the TLR4/NF-κB pathway, suggesting that PPG may be a candidate drug for treating cerebral I/R injury.


The triangular fibrocartilage complex of the wrist--anatomy and function.
The anatomy and function of the triangular fibrocartilage complex (TFCC) of the wrist was studied through anatomic dissections and biomechanical testing of 61 specimens. The TFCC was found to be a homogenous structure composed of, but not dissectable into, the articular disc, the dorsal and volar radioulnar ligaments, the meniscus homologue, the ulnar collateral ligament, and the sheath of the extensor carpi ulnaris. The TFCC was found to be perforated in 53% of specimens dissected, and all of the wrists with a demonstrable perforation showed evidence of damage or erosion of the cartilage of the lunate and/or distal ulna. Biomechanical studies suggest that the TFCC functions both as a cushion for the ulnar carpus and as a major stabilizer of the distal radioulnar joint. Perforations of the TFCC can result in the ulna-lunate abutment and cartilage erosion. Since excision of the TFCC may lead to ulnolunate abutment, chronic wrist pain, and/or instability of the distal radioulnar joint, it is not recommended.


Prognostic significance of serum anti-p53 antibody in patients with hepatocellular carcinoma.
Abnormalities of the p53 gene can lead to the production of anti-p53 antibody in the serum of cancer patients. We evaluated the prognostic significance of anti-p53 antibody in 86 patients with hepatocellular carcinoma (HCC) in comparison with clinicopathological factors: age, sex, etiology, smoking and drinking habits, history of blood transfusion, presence of encephalopathy and ascites, Child classification, Pugh score, bilirubin, albumin, prothrombin time, indocyanine green retention time at 15 min (ICG), underlying liver disease, alpha-fetoprotein (AFP), tumor size, number of tumors, differentiation degree of HCC, presence of extrahepatic metastasis and therapy for HCC. The serum anti-p53 antibody in 86 patients with HCC, 20 with chronic hepatitis (CH) and 20 with liver cirrhosis (LC) was measured by an enzyme-linked immunosorbent assay (ELISA). A single-strand conformation polymorphism-polymerase chain reaction (SSCP-PCR) analysis and loss of heterozygosity (LOH) study of the p53 gene were performed using 8 tissue samples of 8 HCC from four antibody-positive and four antibody-negative patients. The survival probabilities were assessed by the Kaplan-Meier technique, and a Cox regression analysis was used to identify the independent factors for prognosis. Anti-p53 antibody was positive in 32% (28 of 86) of the sera from patients with HCC, but in none of the 20 with CH and 20 with LC. p53 antibody positivity was associated with bilirubin and the number of tumors (p=0.027 and p=0.018, respectively). Overall survival was shorter in the HCC patients with p53 antibody than in those without p53 antibody (p<0.02). Bilirubin, p53 antibody, AFP and ICG were found to be significant prognostic factors by univariate analysis. A Cox multivariate analysis showed that bilirubin and p53 antibody were independent prognostic variables (p<0.0001 and p=0.003, respectively). In four antibody-positive patients, mutation and LOH of the p53 gene were detected in one patient and two patients, respectively. In contrast, only one of four antibody-negative patients exhibited LOH of the p53 gene. Serum anti-p53 antibody could be a useful prognostic factor in patients with HCC.


[Body immunological reactivity in patients with chronic nontumorous diseases of the parotid salivary glands].
Examinations of 81 patients and 30 healthy donors have revealed a variety of immunologic disturbances in different forms of chronic nontumorous conditions of the parotid glands. Lymphocyte sensitization to salivary gland antigens was observed in all the patients, but was the most marked in those with Sjogren's disease. The studies permit considering the detected shifts in the immunity system an important component of the pathogenesis of chronic nontumorous diseases of the parotid glands. These findings may be useful in the differential diagnosis, prognosis, and adequate therapy of salivary gland diseases.


Preparation and characterization of spironolactone-loaded nano-emulsions for extemporaneous applications.
In neonates as well as in adults having swallowing difficulty, oral medication is given through a nasogastric tube making liquid formulations preferable. In this study, we present the high potential of nanometric emulsions formulated by spontaneous surfactant diffusion, as extemporaneous formulations of hydrophobic drug. Spironolactone used as hydrophobic drug model, was incorporated in oil before formulation at a concentration of 13.5mg/g oil. Then, all formulations were evaluated from pharmacotechnical and clinical standpoints, for their use in hospital or community pharmacy. The strength of this new liquid formulation lies on the simplicity, efficiency and reproducibility of their low energy process as on clinical aspects: high dose uniformity, facility to be administered through in nasogastric tube without any retention and a stability of 2 months at least compatible for an extemporaneous use. Moreover, this emulsion presented spironolactone content of 3.75 mg/ml among the most concentrated formulations published.


Management of lacerations and scars.
The purpose of this paper is to review and update the subject of management of scars and lacerations. The surgion who accepts responsibility for management of soft tissue injuries must be aware of fundamental surgical principles as well as detailed technique. Knowledge of basic anatomy and wound physiology is utilized and applied. Emphasis is placed on management of the total pateint. The specialist must accept the role of team captain and establish an order of priority in management and in wound analysis. By having a basic knowledge of wound healing and the lines of favorable contracture, one is better able to prognosticate the eventual healing of the wound after proper management. By being able to predict wound contracture and scar maturation, the patient can be better informed as to what to expect during the period of healing. With lacerations immediate repair is carried out. With scars there is more time for planning and photographic analysis. The contracting forces will by this time have identified themselves and the degree of release of the contracture or camouflage can be determined. Specific wound management emphasized meticulous closure in layers and the use of skin hooks with the interrupted subcuticular suture. Skin sutures with both the continuous subcuticular suture and interrupted sutures of monofilament nylon using the spring-loop are described. Emphasis is placed on the preparation of the skin margins with slight beveling of the skin edges and undermining with precise even thickness of the skin especially at the wound margin. For scar revisions a minimum time of six months should elapse, and 12-18 months is better. Complications include infection, hematoma, wound separation, and rejection of suture materials. Keloids are discussed briefly, particularly regarding the use of intralesional injection steroids. Broken line camouflage techniques are discussed with the regard to breaking up contracture without lengthening. Lengthening either existing or anticipated contractures is accomplished with Z-plasty. The multiple Z-plasty, W-plasty, and Zig-Zag plasty are aimed towards creating a less conspicuous scar and creating some diffusion of the forces of contracture. A technique for a "practical Z-plasty" is described. Both free grafts and skin flaps sometimes must be utilized to fill tissue defects and break up line of contracture. The materials presented and conclusions drawn are based on 25 years of active emergency room coverage and long term follow-up of treated patients. It is the responsibility of the physician to act within the first few hours and to take the time necessary for accurate approximation and realignment of both soft tissue and bone injuries. Minimal scarring depends on accurate approximation of skin margins without tension. The need for early meticulous repair, so that unsightly scars and disfigurements may be prevented, cannot be overemphasized.


Assessment of systolic and diastolic function in asymptomatic subjects using ambulatory monitoring with acoustic cardiography.
Adequately recording diastolic heart sounds and systolic time intervals over longer periods is difficult. Thus, information on the circadian variation of these parameters in an ambulatory population is lacking. Moreover, age-related changes in the prevalence of diastolic heart sounds and measurements of systolic time intervals in an asymptomatic population have not been studied in continuous recordings. Diastolic heart sounds and systolic time intervals will have age and circadian variations that reflect known changes in cardiac function due to aging and circadian rhythms. We studied 128 asymptomatic subjects wearing an ambulatory monitor with acoustic cardiography. The recording spanned a mean duration of 14 hours, including sleep. Data were analyzed for the presence of third (S3) and fourth (S4) heart sounds and for systolic time intervals. In these asymptomatic subjects, S3 was significantly more prevalent in those age <40 years than in those age >40 years, and significantly more pronounced during sleep in the younger group. Also, S4 was significantly more prevalent in those age >40 years and significantly more pronounced during sleep in those age >40 years. In contrast, time intervals reflecting systolic function showed less circadian variation and less worsening with age. The nocturnal increase of S4 in the elderly reflects diastolic impairment-likely a result of changes in diastolic filling patterns with increasing age. An S3 after the age of 40 is a relatively uncommon finding and therefore should be a specific sign of cardiac disease. Continuous monitoring of diastolic heart sounds and systolic time intervals is possible using acoustic cardiography.


Selective CREB-dependent cyclin expression mediated by the PI3K and MAPK pathways supports glioma cell proliferation.
The cyclic-AMP response element binding (CREB) protein has been shown to have a pivotal role in cell survival and cell proliferation. Transgenic rodent models have revealed a role for CREB in higher-order brain functions, such as memory and drug addiction behaviors. CREB overexpression in transgenic animals imparts oncogenic properties on cells in various tissues, and aberrant CREB expression is associated with tumours. It is the central position of CREB, downstream from key developmental and growth signalling pathways, which gives CREB this ability to influence a spectrum of cellular activities, such as cell survival, growth and differentiation, in both normal and cancer cells. We show that CREB is highly expressed and constitutively activated in patient glioma tissue and that this activation closely correlates with tumour grade. The mechanism by which CREB regulates glioblastoma (GBM) tumour cell proliferation involves activities downstream from both the mitogen-activated protein kinase and phosphoinositide 3-kinase (PI3K) pathways that then modulate the expression of three key cell cycle factors, cyclin B, D and proliferating cell nuclear antigen (PCNA). Cyclin D1 is highly CREB-dependent, whereas cyclin B1 and PCNA are co-regulated by both CREB-dependent and -independent mechanisms. The precise regulatory network involved appears to differ depending on the tumour-suppressor phosphatase and tensin homolog status of the GBM cells, which in turn allows CREB to regulate the activity of the PI3K itself. Given that CREB sits at the hub of key cancer cell signalling pathways, understanding the role of glioma-specific CREB function may lead to improved novel combinatorial anti-tumour therapies, which can complement existing PI3K-specific drugs undergoing early phase clinical trials.


Metal analysis in difficult materials with platform furnace Zeeman-atomic absorption spectroscopy. 2. Direct determination of cadmium and lead in milk.
A procedure is described for the direct determination of cadmium and lead in whole milk, skim milk, condensed milk, and human milk. Using a Perkin-Elmer 5000 Z instrument with HGA 500 and L'vov platform and by application of oxygen ashing at approx. 600 degrees C, determination limits of approx. 0.02 microgram/l and 0.7 microgram/l for cadmium and lead, respectively, are attainable. Day-to-day precision is 10% for 0.1 microgram/l of cadmium and 2 micrograms/l of lead. Accuracy control at least at higher levels was possible with DPASV after wet digestion. The contents found with this procedure in cows milk are at the lowest limit of very recent literature data, i.e. on average at 0.05 microgram/l for cadmium and 2 microgram/l for lead for samples from nonpolluted regions. The results indicate that milk does not contribute significantly to heavy metal exposure of man.


Recombinant protein folding and misfolding in Escherichia coli.
The past 20 years have seen enormous progress in the understanding of the mechanisms used by the enteric bacterium Escherichia coli to promote protein folding, support protein translocation and handle protein misfolding. Insights from these studies have been exploited to tackle the problems of inclusion body formation, proteolytic degradation and disulfide bond generation that have long impeded the production of complex heterologous proteins in a properly folded and biologically active form. The application of this information to industrial processes, together with emerging strategies for creating designer folding modulators and performing glycosylation all but guarantee that E. coli will remain an important host for the production of both commodity and high value added proteins.


Determinants of in-hospital mortality in COVID-19; a prospective cohort study from Pakistan.
A prospective cohort study was conducted at the Indus Hospital Karachi, Pakistan between March and June 2020 to estimate the in-hospital mortality among hospitalized COVID-19 patients and its determinants. A total of 170 adult patients were enrolled and all-cause mortality was found to be 39% (67/170). Most non-survivors were above 60 years of age (64%) while gender distribution was quite similar in both groups (males: 77% vs 78%). Most (80.6%) non-survivors came with peripheral oxygen saturation less than 93% while 95% of them had critical disease on arrival. Use of non-invasive ventilation in emergency room was higher among non-survivors (56.7%) versus survivors (26.2%). Median Interleukin-6 levels were higher among non-survivors (78.6: IQR = 33.8-49.0) compared to survivors (21.8: IQR = 12.6-36.3). Most patients in the non-survivor group (86.6%) required invasive ventilator support during hospital stay compared to 7.8% in the survivors. The median duration of ICU stay was longer for non-survivors (9: IQR = 6-12) compared to survivors (5: IQR = 3-7) days. Univariable binary logistic regression showed that age above 60 years, oxygen saturation below 93%, Neutrophil to lymphocyte ratio above 5, procalcitonin above 2ng/ml, unit increase in SOFA score and arterial lactate levels were associated with mortality. We also found that a unit decrease in Pao2/FiO2 ratio and serum albumin were associated with mortality in our patients. Multivariable regression showed that age above 60 years (aOR = 3.4: 95% CI = 1.6-6.9), peripheral oxygen saturation below 93% (aOR = 3.5:95% CI = 1.6-7.7) and serum pro-calcitonin above 2ng/ml (aOR = 4.8; 95% CI = 1.9-12.2) were associated with higher odds of mortality when adjusted by month of admission. Most common cause of death was multisystem organ failure in 35 (56.6%) non-survivors while 22 (35.5%) died due to respiratory failure. Larger prospective studies are needed to further strengthen these findings.


Combined effect of hypertension and hyperuricemia on ischemic stroke in a rural Chinese population.
To investigate the combined effect of hypertension and hyperuricemia to the risk of ischemic stroke in a rural Chinese population. The cross-sectional study was conducted from 2012 to 2013 in a rural area of China. After exclusion for missing data, we finally included 11,731 participants into analysis. After adjusting for age, current smoking, current drinking, BMI, TG, HDL-C and eGFR, hypertension was significantly associated with ischemic stroke in men (OR: 2.783, 95% CI: 1.793, 4.320) and in women (OR: 4.800, 95% CI: 2.945, 7.822). However, hyperuricemia was significantly associated with ischemic stroke only in women (OR: 1.888, 95% CI: 1.244, 2.864). After full adjustment, participants with both hypertension and hyperuricemia had 8.9 times higher risk than those without them. Finally, the interaction between hypertension and hyperuricemia was statistically significant only in women rather than in men after full adjustment. This study demonstrated the positive correlations between hypertension, hyperuricemia and ischemic stroke. Our study also demonstrated the joint effect between hypertension and hyperuricemia towards ischemic stroke only in women, not in men.


Characterization of hospitalized ischemic stroke patients in palestine.
Stroke is a major health problem, yet no studies on stroke have been reported from Palestine. This one-year, hospital-based study was conducted to determine the prevalence of risk factors and the in-hospital mortality rate in patients with ischemic stroke. All patients admitted to Al-Watani government hospital and diagnosed with ischemic stroke between September 2006 and August 2007 were included in the study. Data were obtained by retrospective review of medical charts. Pearson Chi-square and independent t test were used in the univariate analysis. Multiple logistic regression analysis was used to determine the independent predictors of in-hospital mortality rates among the patients. Statistical testing and graphics were carried out using SPSS 15. We identified 153 ischemic stroke patients (83 females and 70 males) of whom 92 were having a first-ever stroke (FES). Patients had several prevalent modifiable risk factors such as hypertension (HTN) (66%), diabetes mellitus (DM) (45.8%), and renal reduced renal function (crcl < 60 ml/ min) (33.9%). Twenty-six (17%) of the patients died during hospitalization. Four variables were significantly associated with in-hospital mortality: history of previous stroke (P= 0.004), crcl at admission (P = 0.004), number of post-stroke complications (P = 0.001), and age (P = 0.043). Multiple logistic regression analysis indicated that the number of post-stroke complications (P= 0.001) and previous stroke (P = 0.03) were significant independent predictors of in-hospital mortality. Screening and better control of risk factors, especially HTN, DM and renal dysfunction, are required to decrease the incidence and in-hospital mortality among patients with ischemic stroke.


Extracellular DNA and Type IV pili mediate surface attachment by Acidovorax temperans.
Extracellular DNA can play a structural role in the microbial environment. Here evidence is presented that an environmental isolate of Acidovorax temperans utilises extracellular DNA for intercellular and cell-surface attachment and that Type IV pili and electrostatic interactions play a role in this interaction. Preliminary attempts to isolate and purify extracellular polysaccharides from A. temperans strain CB2 yielded significant amounts of DNA raising the question of whether this molecule was present as a structural component in the extracellular matrix. The role of DNA in attachment was indicated by experiments in which the addition of DNase to liquid medium inhibited the attachment of Acidovorax to glass wool. A Tn5 insertional mutant, lacking Type IV pili, was unable to initiate attachment. Addition of DNase caused rapid detachment of bound cells, but no detachment occurred when proteinase, RNase or inactivated DNase were used. Addition of MgCl(2) also caused significant detachment, supporting the possible mechanistic role of electrostatic interactions in the attachment process. Although attachment was apparent in early to mid-log phase growth, surprisingly DNA was not detected in the culture supernatant until late stationary phase and coincided with an appreciable loss of cell viability. This suggests that during log-phase growth attachment is mediated by eDNA that is released in low quantities and/or is highly localised within the extracellular matrix and also that stationary phase DNA release through widespread cell lysis may be a separate and unrelated event.


Understanding self-assembly at molecular level enables controlled design of DNA G-wires of different properties.
A possible engineering of materials with diverse bio- and nano-applications relies on robust self-assembly of oligonucleotides. Bottom-up approach utilizing guanine-rich DNA oligonucleotides can lead to formation of G-wires, nanostructures consisting of continuous stacks of G-quartets. However, G-wire structure and self-assembly process remain poorly understood, although they are crucial for optimizing properties needed for specific applications. Herein, we use nuclear magnetic resonance to get insights at molecular level on how chosen short, guanine-rich oligonucleotides self-assemble into G-wires, whereas complementary methods are used for their characterization. Additionally, unravelling mechanistic details enable us to guide G-wire self-assembly in a controlled manner. MD simulations provide insight why loop residues with considerably different properties, i.e., hydrogen-bond affinity, stacking interactions, electronic effects and hydrophobicity extensively increase or decrease G-wire length. Our results provide fundamental understanding of G-wire self-assembly process useful for future design of nanomaterials with specific properties.


Prevalence of biochemical osteomalacia in adults undergoing vitamin D testing.
Prolonged severe vitamin D deficiency can cause osteomalacia, but the 25-hydroxyvitamin D (25OHD) concentration below which this occurs is unknown. We investigated the prevalence of biochemical osteomalacia in adults with a measurement of 25OHD. 25OHD results between 1/1/2009 and 15/6/2020 were obtained from the regional laboratory database, together with measurements of serum calcium, parathyroid hormone (PTH) and alkaline phosphatase (ALP) within 6 months of the index 25OHD. We defined biochemical osteomalacia as all 3 of: albumin-adjusted serum calcium (aCa)<2.0 mmol/L, PTH>7.3 pmol/L and ALP>150 IU/L. Possible osteomalacia was 2/3 criteria with the other test not done. 25OHD measurements associated with significant renal impairment, elevated hepatic transaminases or hypercalcaemia were excluded. 110,046 25OHD measurements were identified over the 11.5 years period. After removal of ineligible measurements, 42,171 25OHD measurements from 32,386 individuals with at least 2 of aCa, PTH and ALP were included in analyses. Median 25OHD was 63 nmol/L; 8% were <25 nmol/L, and 33% were <50 nmol/L. Five index 25OHD measurements met the definition of biochemical osteomalacia, and another 11 were possible osteomalacia. After reviewing available clinical records for these 16 episodes, we classified 9 cases as osteomalacia and 7 as other diagnoses. Thus, the prevalence of biochemical osteomalacia was 0.02% (9/42,171) for 25OHD measurements and 0.23% (8/3432) for 25OHD<25 nmol/L. All cases of osteomalacia with 25OHD measurements prior to supplementation had 25OHD≤18 nmol/L. The prevalence of biochemical osteomalacia is very low, even in individuals with 25OHD<25 nmol/L.