Insights into the Dissolution Behavior of Ledipasvir-Copovidone Amorphous Solid Dispersions: Role of Drug Loading and Intermolecular Interactions.
The generation of a colloidal drug-rich phase by dissolving an amorphous solid dispersion (ASD) is thought to have a positive impact on oral absorption and bioavailability. Thus, understanding which formulations generate these species is important. In this study, ledipasvir-copovidone ASDs, with and without surfactants, were prepared, and their release performance was examined at different drug loadings. An intrinsic dissolution rate assembly was used to limit potential surface area variations among formulations, and the release of both polymer and drug was monitored as a function of time. Drug-rich colloids only formed when the drug loading (DL) was at or below 5%; at a DL of 7.5% or above, drug release became negligible. The drug and polymer released congruently at and below 5% DL and incongruently at higher DLs. Thus, the limit of congruency (LoC) is between 5 and 7.5% DL. X-ray photoelectron spectroscopy (XPS) of partially dissolved tablet surfaces revealed that a drug-rich layer formed on the surface of the tablet. This was most evident for the higher DL ASDs and led to amorphous drug-controlled dissolution. Consequently, the surface drug-enriched layer physically hindered the polymer from further release. Evidence is provided that the extent of drug-polymer interactions as a function of DL plays a central role in dictating the observed release behavior. Some surfactants were found to promote the formation of drug-rich colloids at considerably higher DLs, providing a formulation strategy to increase the LoC.


Immunolocalization of secretory protein-I or chromogranin A in amphibian urinary bladder granular cell granules.
The presence of secretory protein-I (SP-I) or chromogranin A (CGA) in granules isolated from the granular cells of the amphibian urinary bladder epithelium was investigated using ultraimmunohistochemistry. Granules were isolated by cell fractionation using Percoll density gradients. SP-I was isolated and purified from bovine parathyroid glands. Antibodies were raised in rabbits and purified by affinity chromatography. Ultraimmunocytochemistry, employing the avidin-biotin-peroxidase (ABC-complex) procedure, was used to localize SP-I on thin sections of isolated granules. About 27% of the granules from control (-ADH) cells were SP-I+, while 51% of the granules fractionated from hormone treated (+ADH) cells were positive for this protein (p less than 0.0001). Accordingly, granules from ADH-treated cells also showed a significant (p less than 0.0001) increase in total protein.


Encapsulation of cannabinoid drugs in nanostructured lipid carriers.
This study describes the development and optimization of a method to encapsulate the potent and expensive cannabinoids drugs in nanostructured lipid carriers; namely, URB597, AM251 and rimonabant have been considered. NLC production by melt and ultrasonication protocol has been specifically designed to optimize nanoparticle recovery and drug encapsulation efficiency. Special care has been devoted to the modality of oil and water phase emulsification and the entire production has been studied and discussed. NLC recovery, morphology, dimensional distribution and encapsulation efficiency are presented.


Structural determinants of MALT1 protease activity.
The formation of the CBM (CARD11-BCL10-MALT1) complex is pivotal for antigen-receptor-mediated activation of the transcription factor NF-κB. Signaling is dependent on MALT1 (mucosa-associated lymphoid tissue lymphoma translocation protein 1), which not only acts as a scaffolding protein but also possesses proteolytic activity mediated by its caspase-like domain. It remained unclear how the CBM activates MALT1. Here, we provide biochemical and structural evidence that MALT1 activation is dependent on its dimerization and show that mutations at the dimer interface abrogate activity in cells. The unliganded protease presents itself in a dimeric yet inactive state and undergoes substantial conformational changes upon substrate binding. These structural changes also affect the conformation of the C-terminal Ig-like domain, a domain that is required for MALT1 activity. Binding to the active site is coupled to a relative movement of caspase and Ig-like domains. MALT1 binding partners thus may have the potential of tuning MALT1 protease activity without binding directly to the caspase domain.


Human T-cell lymphotropic virus type I and II in transfusion medicine.
As a consequence of migrating populations, IV drug use and, to a lesser extent, blood transfusions, endemic HTLV-I and HTLV-II infections have spread to nonendemic geographic regions. Although the risk that a person infected with HTLV-I will develop significant disease--even over a lifetime--is estimated to be relatively low, our awareness of the serious diseases associated with other retroviruses requires a cautious approach to blood transfusion. Reports from Japan and the United States indicate that programs testing donated blood and excluding units with HTLV-I antibodies have been highly successful in interrupting the spread of HTLV-I by transfusions. One unanticipated outcome of testing large numbers of people in the United States for HTLV-I antibodies has been recognition of the relatively high prevalence of HTLV-II infection, particularly among IV drug users. The long-term effects of HTLV-II infection are also unknown. Until the natural history and clinical consequences of HTLV-II infection are clearly understood, it is only prudent that blood donated by persons identified to be HTLV-II carriers also be excluded.


Behavioral and biochemical effects of intra-accumbens dopaminergic grafts.
6-Hydroxydopamine (6-OHDA) lesions in the ventral tegmental area (VTA) in rats prevented the hyperactivity response to methamphetamine in an open field. Transplantation of mesencephalic dopaminergic cells, obtained from rat embryos, into the nucleus accumbens (NAC) of 6-OHDA-lesioned animals restored the hyperactivity 4 weeks after grafting. By microdialysis of the NAC in freely moving rats no significant differences in baseline concentration of dopamine (DA) among the 3 groups (control, lesioned, grafted) were observed. However, after methamphetamine administration, DA increased significantly during the first 80 min in control animals, during the first 40 min in grafted animals, but did not increase in lesioned animals. On the other hand baseline concentrations of dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) decreased to one sixth to one third of those of controls in 6-OHDA-lesioned animals, and they did not respond to methamphetamine. After grafting, however, DOPAC and HVA restored to control levels and responded to methamphetamine with decreases as was observed in control animals. Data suggest that grafts not only restore the ability to release DA but also improve DA metabolism in the NAC. This might be a reason for recovery of locomotor activity.


The effect of NPWT in wound healing and bacterial count on deep dermal burn injury model: An experimental study.
Sepsis is one of the main causes in burn victim's mortality. The use of negative pressure wound therapy (NPWT) provides an ideal environment to accelerate wound healing. We compare the use of normal saline (NS), intermittent NPWT, continuous NPWT and silver sulfadiazine in wound healing process. This study involved 6 Yorkshire pigs; each pig was induced with 20 burns on the flank area. Burns were divided into 4 treatment groups: NS gauze, intermittent NPWT, continuous NPWT, and silver sulfadiazine dressing. Burns were evaluated on day 1,3,7,14, and 21 for its morphology and bacterial colonization and on day 14 and 21 for the remaining burn surface area. Wound that received NPWT therapy appeared better in both granulation and crust formation. Remaining burn surface area (mm2) on day 14 in NS group, intermittent NPWT, continuous NPWT, and silver sulfadiazine were 107.43 ± 83.43, 178.07 ± 74.83, 146.10 ± 69.1, 126.03 ± 83.22, respectively(p = 0.457); on day 21 in NS group, intermittent NPWT, continuous NPWT, and silver sulfadiazine were 13.16 ± 16.86, 59.49 ± 20.72, 54.79 ± 46.59, 48.95 ± 39.84, respectively(p=0.169). There were no significant differences in each treatment group bacterial colonization(p>0.05). There were no significant correlation between bacterial colonization and remaining burn surface area (p>0.05). While morphologically, the wound in NPWT treatment groups appeared better in granulation and crust formation, the remaining wound surface area and the number of bacterial colonization were not significantly difference compared to standard therapy (silver sulfadiazine and NS gauze). There were no significant correlation between the amount of bacterial colonization and remaining wound surface area on every treatment group.


Relationship between lipolysis, cyclic AMP, and fat-cell size in human adipose tissue during fasting and in diabetes mellitus.
The in vitro relationship between fat-cell size, glycerol release, and peak concentration of cyclic AMP was investigated in human adipose tissue obtained from 25 obese nondiabetic patients before and after a 7-day fast and from 23 patients with untreated diabetes mellitus. In the obese nondiabetic patients there was a linear correlation between fat-cell size and cyclic AMP concentration, and fat-cell size and the rate of lipolysis. This was found both in nonfasting and fasting nondiabetic patients. However, in diabetes mellitus, there was only a relationship between cell size and cyclic AMP concentration. The alpha-adrenergic and beta-adrenergic activity in human adipose tissue was assessed by comparing the effect of isoprenaline and noradrenaline on the cyclic AMP concentration. The activity of both receptors was found to be increased in fasting obese patients and in diabetics. In both conditions the alpha-adrenergic response to catecholamines predominated in small fat cells, whereas in large ones the beta response predominated. The results suggest that during fasting and in diabetes mellitus there is a correlation between fat-cell size and the responsiveness of the adrenergic receptors. Thus, catecholamines may be involved in regulating the fat-cell volume. The view is expressed that the abnormal catecholamine-induced lipolysis is solely due to changes at the level of the adrenergic receptors during fasting, whereas in diabetes mellitus the sequentional activation of lipolysis is disturbed at deeper sites as well.


Antler stem cell-conditioned medium stimulates regenerative wound healing in rats.
When the deer antler is cast, it leaves a cutaneous wound that can achieve scarless healing due to the presence of antler stem cells (ASCs). This provides an opportunity to study regenerative wound healing. In this study, we investigated the therapeutic effects and mechanism of antler stem cell-conditioned medium (ASC-CM) on cutaneous wound healing in rats. In vitro, we investigated the effects of the ASC-CM on proliferation of HUVEC and NIH-3T3 cell lines. In vivo, we evaluated the effects of ASC-CM on cutaneous wound healing using full-thickness skin punch-cut wounds in rats. The results showed that ASC-CM significantly stimulated proliferation of the HUVEC and NIH-3T3 cells in vitro. In vivo, completion of healing of the rat wounds treated with ASC-CM was on day 16 (± 3 days), 9 days (± 2 days) earlier than the control group (DMEM); the area of the wounds treated with ASC-CM was significantly smaller (p < 0.05) than the two control groups. Further molecular characterization showed that the ratios of Col3A1/Col1A2, TGF-β3/TGF-β1, MMP1/TIMP1, and MMP3/TIMP1 significantly increased (p < 0.01) in the healed tissue in the ASC-CM group. In conclusion, ASC-CM effectively accelerated the wound closure rate and enhanced the quality of healing, which might be through transforming wound dermal fibroblasts into the fetal counterparts. Therefore, the ASC-CM may have potential to be developed as a novel cell-free therapeutic for scarless wound healing.


[Occurrence and control of Loxostege stieticatis on host plant Haloxylon ammodendron to Cistanche deserticola].
The investigation of the occurrence and pesticide experiments of Loxostege stieticatis on Haloxylon ammodendron showed that Loxostege stieticatis is an explosive pest; it can be controlled with lower toxic pesticides such as deltametrin; the innocuous comprehension measure should be a good choice to control in field.


Isosteviol acts on potassium channels to relax isolated aortic strips of Wistar rat.
Isosteviol is a derivative of stevioside, a constituent of Stevia rebaudiana, which is commonly used as a noncaloric sugar substitute in Japan and Brazil. In the present study, the role of potassium channels in the vasodilator effect of isosteviol was investigated using potassium channel blockers on isosteviol-induced relaxation of isolated aortic rings prepared from Wistar rats. Isosteviol dose-dependently relaxed the vasopressin (10(-8) M)-induced vasoconstriction in isolated aortic rings with or without endothelium. However, in the presence of potassium chloride (3x10(-2) M), the vasodilator effect of isosteviol on arterial strips disappeared. Only the inhibitors specific for the ATP-sensitive potassium (K(ATP)) channel or small conductance calcium-activated potassium (SK(Ca)) channel inhibited the vasodilator effect of isosteviol in isolated aortic rings contracted with 10(-8) M vasopressin. Also; since the isosteviol-induced relaxation was unchanged by methylene blue, a role of nitric oxide and/or endothelium in the vasodilatation produced by isosteviol could be ruled out. The obtained results indicated that vasodilatation induced by isosteviol is related to the opening of SK(Ca) and K(ATP) channels.


A machine learning-based procedure for leveraging clickstream data to investigate early predictability of failure on interactive tasks.
Early detection of risk of failure on interactive tasks comes with great potential for better understanding how examinees differ in their initial behavior as well as for adaptively tailoring interactive tasks to examinees' competence levels. Drawing on procedures originating in shopper intent prediction on e-commerce platforms, we introduce and showcase a machine learning-based procedure that leverages early-window clickstream data for systematically investigating early predictability of behavioral outcomes on interactive tasks. We derive features related to the occurrence, frequency, sequentiality, and timing of performed actions from early-window clickstreams and use extreme gradient boosting for classification. Multiple measures are suggested to evaluate the quality and utility of early predictions. The procedure is outlined by investigating early predictability of failure on two PIAAC 2012 Problem Solving in Technology Rich Environments (PSTRE) tasks. We investigated early windows of varying size in terms of time and in terms of actions. We achieved good prediction performance at stages where examinees had, on average, at least two thirds of their solution process ahead of them, and the vast majority of examinees who failed could potentially be detected to be at risk before completing the task. In-depth analyses revealed different features to be indicative of success and failure at different stages of the solution process, thereby highlighting the potential of the applied procedure for gaining a finer-grained understanding of the trajectories of behavioral patterns on interactive tasks.


Thoracoscopic right middle lobar bronchus reimplantation for iatrogenic injury in infant: is it a good option?
We report the first case in the literature of thoracoscopic bronchial reimplantation in 5 month-old boy. The child was born with congenital cystic adenomatoid malformation of the right lower lobe associated with lower intralobar pulmonary sequestration diagnosed prenatally. An iatrogenic middle lobar bronchus injury was detected per-operatively during thoracoscopic lobectomy. Management and follow-up were exposed.


Positive predictive value of CD200 positivity in the differential diagnosis of chronic lymphocytic leukemia.
The role of CD200 in the differential diagnosis of chronic lymphocytic leukemia (CLL) and classical mantle cell lymphoma (MCL) is well established. Its role in the differential diagnosis of CLL and other lymphoproliferative disorders (LPD) is less clear, in particular its positive predictive value (PPV). We conducted a systematic review of the use of CD200 in the differential diagnosis of CLL, MCL, and other predominantly leukemic, typically CD103-negative LPD. With the results, we then derived a curve to determine the PPV based on the prevalence of the disorders included in the differential diagnosis. Of 43 publications screened, 27 were included in the systematic review (5,764 patients). The median CD200 positivity rate in all studies and the percentage of CD200-positive (pooled) patients was 100% and 95% (3,061/3,208) in CLL, 4 and 8% (86/1112) in MCL and 56 and 62% (425/689) in other LPD. CD200 is suboptimal for the differential diagnosis of CLL and disorders other than nodal MCL.


Omeprazole and ranitidine, antisecretagogues with different modes of action, are equally effective in causing hyperplasia of enterochromaffin-like cells in rat stomach.
Female rats were treated for 28 days with high doses of the gastric acid secretion inhibitors omeprazole and ranitidine. Omeprazole, which is long-acting, was given orally once daily. Ranitidine, which is short-acting, was given by continuous infusion (via osmotic minipumps, implanted subcutaneously). The aim was to produce a similar degree of acid inhibition with the two drugs. The inhibition of acid secretion over the day and night was more pronounced in the omeprazole-treated rats (maximal inhibition 100%, minimum 85%) than in those receiving ranitidine (mean 70%). In both groups, there was a great increase in plasma gastrin, somewhat greater after omeprazole than after ranitidine. The gastrin concentration in the antrum was almost doubled by both treatments and there was a moderate increase in the number of antral gastrin cells in the omeprazole-treated rats. The number of enterochromaffin-like (ECL) cells (per visual field) increased in the oxyntic mucosa to the same extent (greater than 100%) in the ranitidine- and omeprazole-treated rats. Apart from the gastrin cells in the antrum and the ECL cells in the corpus no other gastric endocrine cell type seemed to respond to treatments with antisecretagogues. We conclude that, regardless of the type of antisecretagogue used, effective and long-term suppression of gastric acid secretion results in sustained hypergastrinemia and increased number of ECL cells. Conceivably therefore, the ECL cell hyperplasia reflects the trophic effect of gastrin.


Interaction of anticancer herbal drug berberine with DNA immobilized on the glassy carbon electrode.
The interaction of anticancer herbal drug berberine with double-strand DNA (dsDNA) and single-strand DNA (ssDNA) in solution, dsDNA immobilized on the glassy carbon electrode prepared by Langmuir-Blodgett technique, were investigated by electrochemical techniques (cyclic voltammetry, differential pulse voltammetry) and UV spectroscopy. The presence of DNA results in a decrease of the currents and a negative shift of the electrode potentials from the DPV curves of berberine, indicating the dominance of electrostatic interactions. The spectroscopy data confirmed that the predominant interaction between berberine and DNA is electrostatic. The binding of berberine with DNA, when analyzed in terms of the cooperative Hill model, yields the binding constant K(a)=2.2(+/-0.2)x10(4) M(-1), corresponding to the dissociation equilibrium constant K(d)=4.6(+/-0.3)x10(-5) M, which in the range of the applied concentrations of DNA (bp) and berberine, and a Hill coefficient m=1.82(+/-0.08) in Britton-Robinson buffer solution (0.05 M, pH 5.72) at T=298 K (25 degrees C). Apparently, at least two molecules of berberine have to bind as a couple to cause, e.g., the "elementary event" of current change. The results are suggestive for further fruitful applications of this anticancer herbal drug and DNA-modified electrodes.


Evaluating the effectiveness of machine learning techniques in forecasting the severity of traffic accidents.
Traffic accidents pose a significant public safety concern, leading to numerous injuries and fatalities worldwide. Predicting the severity of these accidents is crucial for developing effective road safety measures and reducing casualties. This paper proposes an analytic framework that utilizes machine learning models, including Naive Bayes, Random Forest, Logistic Regression, and Artificial Neural Networks, to predict the severity of traffic accidents based on contributing factors. This study analyzed ten years of UK traffic accident data (2005-2014, N = 2,047,256) to develop and compare different ML models. Results show that the proposed Random Forest and Logistic Regression models achieved an 87% overall prediction accuracy, outperforming Naive Bayes (80%) and Artificial Neural Networks (80%). By employing Random Forest-based feature importance analysis, the study identified Engine Capacity, Age of the vehicle, make of vehicle, Age of the driver, vehicle manoeuvre, daytime, and 1st road class as the most sensitive variables influencing traffic accident severity prediction. Additionally, the suggested RF model outperformed most existing models, attaining a remarkable overall accuracy and superior predictive performance across various injury severity classes. The findings have significant implications for developing efficient road safety measures and enhancing the current traffic safety system. The proposed framework and models can be adapted to various datasets to achieve accurate and effective predictions of traffic accident severity, serving as a valuable reference for implementing traffic accident management and control measures. Future research could extend the proposed framework to datasets containing Casualty Accident information to further improve the accuracy of injury severity prediction.


Knowledge, attitudes and beliefs towards e-cigarettes among e-cigarette users and stop smoking advisors in South East England: a qualitative study.
Aim To explore how e-cigarettes are perceived by a group of e-cigarette users and a group of Stop Smoking Advisors (SSAs), what are the risks and benefits they associate with e-cigarettes and how do these understandings shape participants' attitude towards e-cigarettes? Face-to-face and phone interviews were conducted with 15 e-cigarette users and 13 SSAs in South East England between 2014 and 2015. Transcribed data were analysed inductively through thematic analysis. Findings E-cigarettes were used as a therapeutic aid to stop or cut down smoking and as a smoking substitute. A prominent theme is the uncertainty e-cigarettes have generated. This included ambiguity of e-cigarettes' status and efficacy, and ambiguity of e-cigarettes' physical and social risks. Different attitudes towards e-cigarettes were identified. E-cigarettes' benefits and risks should be continuously evaluated, put into perspective and circulated to avoid ambiguity. Stop smoking services need to recognise the benefits that can be gained by using e-cigarettes as a harm reduction tool.


Analysis of human nasal mucous glycoproteins.
Human nasal turbinates were cultured in the presence of 3H-glucosamine, which is incorporated into nasal mucous glycoproteins. Nasal mucous glycoprotein was then characterized biochemically, and the effects of various neurohormones and immunologic stimulation on mucous glycoprotein release were analyzed. Fractionation of nasal mucous glycoprotein by gel filtration chromatography revealed a molecular size range of 2 to 200 X 10(5) (as judged by protein markers) but displayed a single, acidic charge, as reflected both in a narrow elution pattern from DEAE-cellulose and a sharp isoelectric focusing point of 2.6. Highly enriched nasal mucous glycoprotein preparations consisted of 80 per cent carbohydrate and 20 per cent protein (by weight) and included enzymatically cleavable carbohydrate side chains with molecular weights of 1,600 to 1,800. Thus, nasal mucous glycoproteins are a family of molecules that express uniform acidic charge characteristics and a wide range of molecular sizes. Cholinergic stimulation of atropine-inhibitable muscarinic receptors increased nasal mucous glycoprotein release in a dose-related manner, as did alpha-adrenergic stimulation. However, beta-adrenergic stimulation did not affect mucous glycoprotein release. Immunologic stimulation of nasal mast cells by either reversed anaphylaxis or antigen challenge after passive sensitization caused both histamine release and increased mucous glycoprotein release. Thus, nasal turbinates provide an accessible source of tissue for the analysis of nasal mucus secretion and mast cell degranulation and may provide a model for the study of pharmacologic approaches to the universally experienced discomfort of rhinorrhea.


Bacterial community dynamics and metabolic functions prediction in white button mushroom (Agaricus bisporus) during storage.
White button mushroom (Agaricus bisporus) is rich in nutritional value, but it is easily infected by microorganisms during storage, which leads to spoilage and shortens the storage time. In this paper, A. bisporus at different storage times was sequenced by Illumina Novaseq 6000 platform. QIIME2 and PICRUSt2 were used to analyze the changes of bacterial community diversity and predict metabolic functions during storage of A. bisporus. Then, the pathogenic bacteria were isolated and identified from the spoilt samples of A. bisporus with black spot. The results showed that the bacterial species richness of A. bisporus surface gradually decreased. 2,291 ASVs were finally obtained through DADA2 denoising, belonging to 27 phyla, 60 classes, 154 orders, 255 families and 484 genera. The abundance of Pseudomonas on the surface of fresh A. bisporus sample was 22.8%, which increased to 68.7% after 6 days of storage. The abundance significantly increased and became a dominant spoilage bacterium. In addition, A total of 46 secondary metabolic pathways belonging to 6 categories of primary biological metabolic pathways were predicted during storage of A. bisporus, and metabolism (71.8%) was the main functional pathway. Co-occurrence network analysis revealed that the dominant bacterium Pseudomonas was positively correlated with 13 functional pathways (level 3). A total of 5 strains were isolated and purified from diseased A. bisporus surface. The test of pathogenicity showed that Pseudomonas tolaasii caused serious spoilage of A. bisporus. The study provided a theoretical basis for the development of antibacterial materials to reduce related diseases and prolong the storage time of A. bisporus.


Intracellular cytokine detection by flow cytometry in pigs: fixation, permeabilization and cell surface staining.
Intracellular flow cytometry is a method of cytokine detection that allows simultaneous detection of intracellular cytokines and cell surface markers. This important method is not extensively used in pigs, in particular due to the inaccessibility of proper methodological protocols modifying comprehensive human protocols. The aim of this study was to find the best procedure for fixation and permeabilization of porcine blood leukocytes and simultaneous cell surface staining. Permeabilization with commercial kits gave better results in most of the chosen parameters compared with combinations of different concentrations of paraformaldehyde and saponin. Among the commercial kits tested, the best results were obtained with the IntraStain kit. Cell surface markers were detected on cells stimulated for cytokine production by antibodies anti-CD14 (clone MIL-2), anti-SWC3, anti-CD4 and anti-CD8 except anti-CD14 (clone Tük4). While anti-CD8 (clone MIL-12) must be used for staining of unfixed cells, the other antibodies recognize fixed and/or permeabilized cells. Moreover, anti-SWC3 and anti-CD14 (clone MIL-2) antibodies can stain cells during the permeabilization step. These modifications of the cell surface staining protocol allow the researcher to speed up the procedure of intracellular cytokine staining or to combine cell surface staining and intracellular cytokine staining. The present study can serve as a particular protocol of intracellular cytokine detection and as a suggestion for optimization of the fixation, permeabilization and cell surface staining procedure in any laboratory.


A utility analysis of drinking.
In a study applying a utility analysis to drinking behavior, a questionnaire on beliefs about the effects of alcohol and the desirability of those effects was administered to a sample of college students and a sample drawn from the population. Measures of the expected utility of drinking were calculated. Utility scores were positively related to drinking habits, due to heavier drinkers' increased expectations of positive consequences of drinking and more positive evaluation of all consequences. It is suggested that studies on expectancies about drinking should include not only measures of behavioral expectations but measures of the desirability of alcohol effects.


Electro-oculography in bilateral optic neuropathy.
Effect of bilateral, optic neuropathy on the function of retinal pigment epithelium has not been investigated extensively to date. This study aimed to determine the effect of bilateral, optic neuropathy on light peak:dark trough ratio, light peak and dark trough values of electro-oculography. Thirty-seven patients with a clinical diagnosis of bilateral optic neuropathy and 40 control subjects were recruited in this observational, cross-sectional study, carried out at the Neurophysiology unit, Teaching Hospital, Peradeniya, Sri Lanka. Pattern reversal visual evoked potentials, pattern electroretinography and electro-oculography were recorded in all of them. Twenty-four patients (64.9%) had reduced light peak:dark trough ratio values. The median light peak:dark trough ratio ± inter quartile range of the patient group (1.50 ± 0.4) was significantly lower than that of the controls (1.80 ± 0.2), (p < 0.001). Median dark trough value was significantly higher in patients compared to the control value. These changes may be due to higher conductance of ion channels in the retinal pigment epithelium, sub retinal space creation, changes in choroidal circulation or secondary to photoreceptor abnormalities in the macular region. These results indicate that retinal pigment epithelium might be affected in bilateral optic neuropathy.


Epigenetic age acceleration of early stage hepatocellular carcinoma tightly associated with hepatitis B virus load, immunoactivation, and improved survival.
Properly stratifying high-risk individuals with early stage hepatocellular carcinoma (HCC) is essential to identify patients in which the potentially therapies can be offered. To this context, we systematically investigated the prognostic value of epigenetic clock with early stage HCC as well as the association with other molecular characteristics. We computed DNA methylation (DNAm) age of 256 early stage HCC patients and 50 normal samples from TCGA by Horvath clock model. The characteristics of two DNAm age subgroups were differentiated regarding HBV expression, pathway activity, epigenomic, and genomic alteration. Cox regression and restricted cubic spline (RCS) analysis were utilized to evaluate the prognostic value of epigenetic acceleration. DNAm age was significantly associated with chronological age in normal tissue but largely disrupted in tumors (P< .001), and showed significant negative correlation with HBV expression (P< .05). We identified two DNAm age groups (DNAmAge-ACC and DNAmAge-DEC), and the former presented with an immunoactive phenotype (all FDRs<0.05 in enrichment analysis), CpG island hypermethylation (P< .001), and lower mutation burden (P= .018). Every 10-year increase in DNAm age was associated with a 18% decrease in fatality after adjustment for major clinical variables; DNAmAge-ACC had 50% lower mortality risk than DNAmAge-DEC (HR: 0.50, 95% CI: 0.27-0.94, P= .03). RCS revealed the fatality risk significantly decreased as epigenetic age accelerated (P = .04). Conclusions. In summary, we highlighted the prognostic value of epigenetic age acceleration for early stage HCC; better prognosis, relatively lower HBV load, and higher enrichment of immune signatures were tightly associated with epigenetic age accelerated tumors.


Autoradiography of hypothalamic regions after acute hyperosmolarity in mice.
Many regions around the third ventricle of the brain are thought to participate in the regulation of water intake. In this study we have visualized in acutely hyperosmolar mice the hypothalamic regions by using autoradiography and [14C]deoxyglucose as marker. By applying a recently published, inexpensive, calibrated photographic method for the analysis of the autoradiographs, we can show that the medial parts of the hypothalamus near the third ventricle increased their uptake of deoxyglucose in comparison with the lateral parts. Densitometric measurements confirmed the results. The results also indicate that parts of the medial hypothalamus, even those that are more caudal than the paraventricular nuclei, react to hyperosmolarity.


Familial erythrophagocytic lymphohistiocytosis: surface marker analysis using monoclonal antibodies.
Surface marker analysis of atypical cells, present at elevated levels in the peripheral blood of a 2-yr-old patient with familial erythrophagocytic lymphohistiocytosis (FEL), was performed with monoclonal antibodies. The atypical cells expressed T11, T8, Ia and LeuM2 antigens, markers of suppressor/cytotoxic T lymphocyte and monocyte. The monoclonal antibody-defined surface markers were thus distinguishable from those of other lymphoproliferative disorders clinically resembling FEL.


First Report of New Delhi Metallo-β-Lactamase-6 (NDM-6) in a Clinical Acinetobacter baumannii Isolate From Northern Spain.
The objective of this study was the phenotypic and genotypic characterization of a carbapenem resistant Acinetobacter baumannii (CRAB) isolate. The isolate, recovered in Northern Spain in 2019, was identified by MALDI-TOF to the species level. Antimicrobial susceptibility testing was performed using the Phoenix BD NMIC-502 Panel, E-test, and broth microdilution methods. The presence of a metallo-β-lactamase (MBL) was verified by PCR and immunochromatographic assays. The genetic location of the MBL was confirmed using S1-pulsed-field gel electrophoresis (S1-PFGE) followed by Southern blot hybridization. Whole genome sequencing (WGS) was completed using the Miseq and MinION platforms, followed by core-genome MLST (cgMLST) and seven-locus MLST analysis. The CRAB was assigned ST85 (Pasteur scheme) and ST957 (Oxford scheme) representing international clone (IC) 9 and harbored the intrinsic β-lactamase OXA-94 with ISAba1 upstream of it, and the MBL bla NDM-6. Hybridization experiments revealed that the bla NDM-6 was encoded on the chromosome. Using WGS the bla NDM-6 environment could be identified arranged in the following order: ISAba14, aphA6, ISAba125, bla NDM-6, ble MBL, trpF, dsbC, cutA, and ISAba14. Downstream, a 10,462 bp duplication was identified, including a second copy of bla NDM-6 in the following genetic composition: ISAba125, bla NDM-6, ble MBL, trpF, dsbC, cutA, and ISAba14. To our knowledge, this is the first description of bla NDM-6 in A. baumannii. The MBL was present in two copies in the chromosome in a new genetic environment associated with IS elements highlighting the contribution of mobile genetic elements in the dissemination of this gene.


Issues and controversies in the management of cleft palate.
Cleft palate management is complex. There is no current agreement on the appropriate treatment strategy. Extensive disagreement on the pathophysiology, timing of intervention, and techniques of surgical repair have added to the confusion. To provide a comprehensive guide to the management of cleft palate is difficult. However, several main points should be emphasized. Normal speech should be the most important consideration in the therapeutic plan. Growth disturbance should be minimized, but not at the expense of speech impairment, because the facial distortion can be satisfactorily managed with further surgery, whereas speech impairment can often be irreversible. We believe repair of cleft palate to establish a competent velopharyngeal sphincter should be completed from 6 to 12 months of age. This is done early enough to minimize the development of an often irreversible pathologic compensatory speech pattern, but late enough not to increase significantly the surgical risk to the infant. Surgical interventions should be designed to cause minimal disruption of the palate, to decrease the severity of subsequent growth problems. There is a need for well-controlled, prospective studies to establish the validity of the widely different claims of superior results from various techniques. We believe strongly that cleft patients should be managed in a center with a multidisciplinary team. The benefits of these teams have been elaborated. Cleft palate embodies one of the major tenets of plastic surgery, the achievement of an aesthetic result with minimal interference with function. Cleft palate remains a significant and interesting challenge for current and future plastic surgeons.


Modeling the growth of diverse microorganisms during feast-famine enrichment.
Polyhydroxyalkanoates (PHAs) are biodegradable polymers that can decrease the severe environmental pollution of petroleum plastics. PHA production by mixed microbial communities has been extensively studied to lower the high PHA prices. However, the competition between distinct microbial communities during the enrichment of PHA accumulators in mixed cultures has not been widely investigated. Thus, in this work, we developed a mathematical model for the competition between PHA accumulators and non-PHA accumulators in the feast-famine enrichment strategy. The developed model successfully simulated published lab-scale experimental data for Plasticicumulans acidivorans, a well-studied PHA accumulator that can store PHA up to 90% of the cell weight. The growth kinetics for both PHA and non-PHA accumulators were estimated and compared to the values in the literature. The uncertainties in the model kinetics were studied by expanding the model to include additional sub-biomass components for each heterotrophic group. As a result, the microbial diversity of microbial communities was observed to influence the enrichment of PHA accumulators in mixed cultures. Additionally, the calibrated model was applied to investigate the cultivation conditions, such as cycle lengths, carbon-to-nitrogen ratio, and solids retention time for successful P. acidivorans enrichment in mixed cultures. The developed model can be applied to control the cultivation and enrichment of PHA accumulators in large-scale PHA production systems. PRACTITIONER POINTS: A new model for the enrichment of PHA accumulators was developed. The model can simulate PHA accumulation by enriched cultures. The model was calibrated and validated for Plasticicumulans acidivorans. The impact of microbial diversity on enriching PHA accumulators was investigated. Short cycles (<12 h) and SRT (<10 d) are suggested for successful enrichment.


Trichloroacetic acid (TCA) precipitability of 125I in the blood of mice fed 125I.
A recent study showed that after feeding 125I-labelled proteins or free 125I to mice, as much as 40% of total radioactivity in the circulation precipitated upon mixing whole blood with 10% trichloroacetic acid. We examined this potential limitation to the use of radiolabelled tracers for studies on intestinal digestion of proteins and protein uptake, and identified its mechanism. BALB/c mice were gavage-fed or injected intravenously (i.v.) with Na125I. Blood obtained at 15 min was added directly to 10% trichloroacetic acid (TCA) or was processed to obtain serum or plasma. On mixing with 10% TCA, 25-33% of the radioactivity in whole blood was precipitated; less than 2% of the radioactivity in plasma or serum was precipitated. In vitro studies identified hemoglobin as the primary carrier protein participating in this reaction. If hemoglobin was replaced by methemoglobin or cyanomethemoglobin, then the reaction with 125I did not occur, suggesting that iron in the heme group may be the site for 125I binding and that iron must be in its reduced or ferrous form (Fe2+). The administration of non-radioactive NaI in vivo or its addition to reaction mixtures in vitro completely inhibited the precipitation of 125I by hemoglobin in the presence of TCA. Thus the addition of non-radioactive iodide to TCA stock solutions may effectively prevent non-specific binding of 125I to free hemoglobin released unintentionally during venipuncture or at other stages of blood processing.


Lysosomal enzymes in tear fluids from patients with Terrien's marginal corneal degeneration.
We examined biochemically the lysosomal enzyme activities in tear fluids from patients with mild myopia, senile cataract, and Terrien's marginal corneal degeneration. Tear acid phosphatase activities in Terrien's degeneration were almost the same as those in mild myopia and senile cataract, while those of N-acetyl-beta-D-glucosaminidase in Terrien's degeneration were higher. The high activity of tear N-acetyl-beta-D-glucosaminidase may be derived from the lacrimal gland and infiltrate histiocytelike cells in Terrien's marginal corneal degeneration.


CD studies on the reversed heme orientation in monomeric Glycera dibranchiata hemoglobins.
Circular dichroism spectra of three monomeric components of Glycera dibranchiata hemoglobins are reported. Contrary to what is found for most hemoglobins and myoglobins, G. dibranchiata hemoglobins display largely negative dichroic spectra in the Soret region. Independent NMR measurements have shown that the same monomeric hemoglobin components contain the heme moiety predominantly (greater than 85%) oriented in a reversed way with respect to the orientation which occurs in most hemoglobins and myoglobins. On the basis of these independent NMR studies, and also of previous data on other invertebrate hemoproteins, a correlation appears evident between reversed heme orientation in hemoglobins and negative ellipticity in the Soret CD spectrum. This represents a simple tool to evaluate this aspect of heme asymmetric environment.


The role of the septal nuclei in the release of acetyl-choline from the rabbit cerebral cortex and dorsal hippocampus and the effect of atropine.
Acetylcholine (ACh) was collected from the alvear surface of the dorsal hippocampus and cerebral cortex in chloralose-urethane anaesthetized or unanaesthetized rabbits. With anaesthesia, the resting release of ACh from the hippocampus was greater than that from the cortex. Wthout anaesthesia, the resting release from both areas was much higher and very similar. The addition of atropine sulphate (1 microgram/ml) to the collecting fluid or the administration of Artane (2 mg/kg i.v.) increased resting ACh release from both the hippocampus and cortex to similar output levels. Atropine also increased ACh release due to stimulation of the medial septum (MS) or mesencephalic reticular formation (MRF). Removal of the septum abolished the effect of atropine on resting ACh release and on release evoked by MRF stimulation from both the hippocampus and cortex. The data indicate that the septum is an essential pathway for cholinergic fibres ascending to the cerebral cortex and hippocampus. They also demonstrate that the septal cholinergic fibres must be intact and active for atropine to increase ACh release from their terminals.


Infusion of hypertonic saline (7.5% NaCl) causes minor immunological changes in normovolaemic women.
Haemorrhagic shock is treated effectively by infusion of hypertonic saline/colloid solutions. Furthermore, previous studies found hypertonicity to affect immune responses in animals and in human blood cell cultures. It is unknown, however, whether hypertonic saline infusion affects immune responses in humans. In a randomized double-blind study, we infused 4 ml kg-1 of 7.5% NaCl or 0.9% NaCl over 10 min in 20 fasting women before hysterectomy. We collected peripheral blood at baseline, 30, and 120 min after start of the infusion and before surgery for the determination of leucocyte and differential count; lymphocyte subtypes; neutrophil chemotaxis; elastase concentration; and the cytokine's tumour necrosis factor-alpha (TNFalpha), interleukin (IL)-1beta, IL-6, IL-8, IL-1 receptor antagonist (IL-1ra), and IL-10. Phytohaemagglutinin (PHA)-induced lymphocyte proliferation and natural killer cell cytotoxicity were measured before and 120 min after infusion. Compared with normal saline, infusion of hypertonic saline temporarily increased the number of B cells in peripheral blood (P < 0.01); increased the concentration of plasma elastase, a marker of neutrophil degranulation (P < 0.05); and decreased the number of circulating neutrophils (P < 0.001). No other effects were detected in the measured immunological parameters. The immunological consequences of hypertonic saline infusion seem to be modest and are unlikely to cause any clinical effects, at least in normovolaemic women.


A systematic study of independently-tuned room-specific PBS beam model in a beam-matched multiroom proton therapy system.
In the existing application of beam-matched multiroom proton therapy system, the model based on the commissioning data from the leading treatment room was used as the shared model. The purpose of this study is to investigate the ability of independently-tuned room-specific beam models of beam-matched gantries to reproduce the agreement between gantries' performance when considering the errors introduced by the modeling process. Raw measurements of two gantries' dosimetric characteristics were quantitatively compared to ensure their agreement after initially beam-matched. Two gantries' beam model parameters, as well as the model-based computed dosimetric characteristics, were analyzed to study the introduced errors and gantries' post-modeling consistency. We forced two gantries to share the same beam model. The model-sharing patient-specific quality assurance (QA) tasks were retrospectively performed with 36 cancer patients to study the clinical impact of beam model discrepancies. Intra-gantry comparisons demonstrate that the modeling process introduced the errors to a certain extent indeed, which made the model-based reproduced results deviate from the raw measurements. Among them, the deviation introduced to the IDD curves was generally larger than that to the beam spots during modeling. Cross-gantry comparisons show that, from the beam model perspective, the introduced deviations deteriorated the high agreement of the dosimetric characteristics originally shown between two beam-matched gantries, but the cross-gantry discrepancy was still within the clinically acceptable tolerance. In model-sharing patient-specific QA, for the particular gantry, the beam model usage for intensity-modulated proton therapy (IMPT) QA plan generation had no significant effect on the actual delivering performance. All reached a high level of 95.0% passing rate with a 3 mm/3% criterion. It was preliminary recognized that among beam-matched gantries, the independently-tuned room-specific beam model from any gantry is reasonable to be chosen as the shared beam model without affecting the treatment efficacy.


Effect of normobaric hyperoxia on antioxidant defenses of HeLa and CHO cells.
The effect of increased intracellular oxygen activation on cellular antioxidant defenses in CHO and HeLa cells was studied. In both cell types, hyperoxic exposure (up to 4 days, 600-700 mm Hg O2) and in CHO cells menadione (up to 3 days, 15 microM) failed to affect the enzymatic antioxidant defenses Mn-containing superoxide dismutase (Mn-SOD), CuZn-SOD, catalase and glutathione peroxidase. The markedly increased antioxidant enzyme activities observed in a recently obtained oxygen-tolerant CHO variant persisted under normoxia. These data suggest that the synthesis of antioxidant enzymes is constitutive. Glutathione levels of HeLa cells did not respond to hyperoxia whereas in CHO cells hyperoxia and menadione exposure resulted in a 2- and 7-fold increase in glutathione contents, respectively. However, considering the large variations in glutathione contents observed under normal culture conditions, it is uncertain whether this increase is to be considered as a true adaptive response.


Evaluating the efficacy of statins and ACE-inhibitors in reducing gastrointestinal toxicity in patients receiving radiotherapy for pelvic malignancies.
3-Hydroxy-methylglutaryl coenzyme-a reductase inhibitors (statins) improve survival following pelvic irradiation for cancer. Large studies suggest that patients with hypertension may have reduced gastrointestinal (GI) toxicity. Animal data suggest that statins and ACE inhibitors (ACEi) may protect against normal tissue injury. Their efficacy in humans has not been reported. To evaluate the impact of statins and ACEi on normal tissue toxicity during radical pelvic radiotherapy. GI symptomatology was recorded prospectively before radiotherapy, weekly during treatment and 1 year later using the inflammatory bowel disease questionnaire-bowel (IBDQ-B) subset. Cumulative acute toxicity (IBDQ-B AUC) and worst score were determined. Dose, brand and duration of statin and/or ACEi usage were obtained from General Practitioners. Of 308 patients recruited, 237 had evaluable acute drug and toxicity data and 164 had data at 1year. Acutely, 38 patients (16%) were taking statins, 39 patients (16.5%) were taking ACEi and 18 patients (7.6%) were taking statin+ACEi. Mean changes in acute scores were 7.3 points (non-statin users), 7.3 (non-ACEi users) and 7.0 (non-statin+ACEi users) compared to 4.8 points (statin users), 5.0 points (ACEi users) and 4.9 points (statin+ACEi users). Statin use (p=0.04) and combined statin+ACEi use (p=0.008) were associated with reduced acute IBDQ-B AUC after controlling for baseline scores (ANOVA). At 1 year, users maintained higher IBDQ-B scores than non-users in all user subgroups. Use of statin or statin+ACEi medication during radical pelvic radiotherapy significantly reduces acute gastrointestinal symptoms scores and also appears to provide longer-term sustained protection.


Manipulating the avian epiblast and epiblast-derived stem cells.
Compared to eutherian mammals, birds retain a primitive form of epiblast development. Molecular studies of the avian epiblast can provide valuable insight for mammalian epiblast research. Here, we introduce several basic techniques in handling epiblast-stage embryos of the chick, the major model organism for avian developmental biology studies. We describe how to collect embryos for RNA extraction and gene expression analysis, to set up ex ovo New culture for overexpression, bead graft and small molecule-based inhibitor studies, and to carry out whole-mount RNA in situ hybridization analysis. We introduce a novel and simple method for molecular perturbation of the epiblast differentiation in ovo. We also describe how to perform primary chicken epiblast cell culture, to establish stable epiblast stem cell (Epi-SC) lines, and to assay for pluripotency in primary epiblast cells and Epi-SCs.


MitoQ protects against high glucose-induced brain microvascular endothelial cells injury via the Nrf2/HO-1 pathway.
Brain microvascular endothelial cells (BMECs) dysfunction is related to the pathogenesis of neurovascular complication of diabetes mellitus that adversely lead to various CNS disorders. Mitoquinone (MitoQ) is a mitochondria targeted antioxidant that exerts multiple protective effects in many oxidative damage-related diseases. In this study, we determined the protective effects of MitoQ on high glucose (HG)-induced BMECs injury and investigated the underlying mechanism. We found that HG significantly reduced the expression of Nrf2 and HO-1, decreased mitochondrial membrane potential, increased intracellular and mitochondrial reactive oxygen species (ROS) generation, induced cytoskeletal damage and apoptosis in BMECs. In addition, Mito tempol, a mitochondrial ROS scavenger, significantly reduced HG-induced mitochondrial ROS production and attenuated cytoskeletal damage and cell apoptosis, suggesting MtROS production was involved in HG-induced BMECs injury. Moreover, we found that MitoQ treatment significantly upregulated the expression of Nrf2 and HO-1 in HG-induced BMECs, which is accompanied by improved mitochondrial membrane potential and decreased MtROS production. Meanwhile, MitoQ treatment also remarkably attenuated HG-induced cytoskeletal damage and cell apoptosis in BMECs. However, inhibitor of Nrf2 with ML385 impaired the protective effects of MitoQ in HG-induced BMECs. In conclusion, our results suggest that MitoQ exerts protective effect on HG-induced BMECs injury via activating Nrf2/HO-1 pathway.


Auditory brainstem implants in NF2 patients: results and review of the literature.
Neurofibromatosis Type 2 (NF2) patients have multiple central nervous system tumors and, specifically, bilateral vestibular schwannomas (VSs) causing bilateral deafness. If the cochlear nerve is not preserved during tumor removal, the only hearing rehabilitation in these patients could be via an auditory brainstem implant (ABI). Retrospective case study and literature review. Tertiary referral cranial base center. In 24 NF2 patients, 25 ABIs were placed in the lateral recess of the fourth ventricle after VS surgery via a translabyrinthine approach. In this series, a large range of results are observed: from open speech and use of the telephone to no ABI use, because of the poor sound identification ability. Of the 24 patients, 19 use their ABI on a daily basis, 4 are nonusers, and 1 died of NF2 progression. A multivariate analysis did not reveal a good predictor for ABI outcome. In literature, the results of ABI in NF2 are difficult to compare, and the overall outcome was poor compared with cochlear implantation results. Auditory brainstem implantation in NF2 patients directly after tumor removal is a safe procedure and the best means of hearing rehabilitation if the cochlear nerve is not preserved. The results in NF2 cases in the literature and these series are poor compared with cochlear implantation. If a cochlear implant is possible, it has the preference over an ABI, also in NF2. Nevertheless, the majority of the patients have benefit of the ABI during daily life particularly in combination with lip reading.


A combined extract of Cinnamomi Ramulus, Anemarrhenae Rhizoma and Alpiniae Officinari Rhizoma suppresses production of nitric oxide by inhibiting NF-kappaB activation in RAW 264.7 cells.
An herbal mixture prepared with Cinnamomi Ramulus, Anemarrhenae Rhizoma and Alpiniae Officinari Rhizoma (CAA) is used in oriental medicine for treating several ailments. The purpose of this study was to determine the mechanisms by which CAA elicits an antiinflammatory effect on nitric oxide (NO) production in the mouse macrophage cell line RAW 264.7 cells. The results indicated that lipopolysaccharide (LPS)-induced NO production was inhibited by CAA in a dose-dependent manner. Western blotting and RT-PCR analysis demonstrated that CAA decreased LPS-induced inducible nitric oxide synthase (iNOS) protein and gene expression in RAW 264.7 cells. Furthermore, CAA inhibited the LPS-induced DNA binding activity of nuclear factor-kappa B (NF-kappaB) and this effect was mediated through inhibiting the degradation of inhibitory factor-kappaBalpha (IkappaBalpha). Therefore, the results demonstrate that CAA inhibits LPS-induced production of NO and expression of iNOS by blocking NF-kappaB activation. CAA might be a potential therapeutic candidate for treating inflammatory diseases such as arthritis.


Retraction: Tumor Microenvironment in Oral Cancer Following Neoadjuvant Pembrolizumab: Preliminary Analysis of the Histopathologic Findings.
[This retracts the article DOI: 10.3389/froh.2021.653104.].


Erratum to "Carotid computed tomography angiography after cobalt-based alloy carotid artery stenting using ultra-high-resolution computed tomography with model-based iterative reconstruction" [Radiol Case Rep 2021;16:3721-8].
[This corrects the article DOI: 10.1016/j.radcr.2021.09.003.].


Interaction of zinc with ionotropic and metabotropic glutamate receptors in rat hippocampal slices.
The actions of zinc on ionotropic and metabotropic glutamate receptors were studied using intracellular recording in acutely prepared adult rat hippocampal slices and in organotypic hippocampal slice cultures. In control Krebs, glutamate and non-N-methyl-D-aspartate (NMDA) agonist-induced responses were enhanced by zinc (25-300 microM). However, under conditions favouring NMDA receptor activation, zinc inhibited glutamate- and NMDA-induced responses. Metabotropic glutamate receptor-mediated responses activated in cultured slices by 1-amino-cyclopentane-1,3-dicarboxylate (1S,3R-ACPD) or by quisqualate, were reversibly inhibited by zinc (200 microM). These results indicate that zinc can inhibit responses induced by activation of metabotropic glutamate receptors and reaffirm that zinc has a differential effect on NMDA and non-NMDA receptors.


Voltage-dependent ion channel formation by rigid rod-shaped polyols in planar lipid bilayers.
In this Letter, we describe the appearance of large, voltage-dependent currents in BLM induced by rigid rod-shaped polyols that function without charge and permanent dipole moment. The capacity of these symmetrical, nonpeptide models to form either short-living nanopores or small ion channels is shown to depend critically on the length of rigid-rod scaffold as well as the nature of the lateral side chains.


Identification in chickens of an evolutionarily conserved cellular ets-2 gene (c-ets-2) encoding nuclear proteins related to the products of the c-ets proto-oncogene.
In chicken cells, we previously identified a set of proteins (p58-64) structurally related to, but distinct from, the products encoded by the c-ets proto-oncogene. We report here the isolation and nucleotide sequence of a cDNA encoding nuclear products of mol. wt 58, 60, 62 and 64 kd, indistinguishable from those detected in chicken cells. The p60 and p64 species appear to represent phosphorylated versions on serine and threonine residues of p58 and p62. The homology of p58-64 to other ets-related proteins, including the v-ets encoded domain of the transforming protein of avian leukemia virus E26 and p54c-ets, the translation product of the chicken (Ck) c-ets gene, is confined to two regions of 175 and 96 amino acid residues localized respectively at the carboxy-terminal domain and close to the amino-terminal domain of these molecules. This cDNA corresponds to a gene localized in a locus distinct from that of c-ets which is transcribed as a 4.0-kb RNA species in most chicken tissues. We also identified the human (Hu) c-ets-2-encoded products as two proteins of 60 and 62 kd, highly related to chicken p58-64. This, together with the fact that the amino acid sequence of the cDNA encoding p58-64 is 95% identical to the reported partial sequence of a Hu-c-ets-2 cDNA, indicates that p58-64 are the translation products of the Ck-c-ets-2 gene.


Effect of simvastatin in patients with aneurysmal subarachnoid hemorrhage: A systematic review and meta-analysis.
Simvastatin might be beneficial to the patients with aneurysmal subarachnoid hemorrhage. However, the results remained controversial. We conducted a systematic review and meta-analysis to explore the efficacy of simvastatin for aneurysmal subarachnoid hemorrhage. PubMed, EMbase, Web of science, EBSCO, and Cochrane library databases were systematically searched. Randomized controlled trials (RCTs) assessing the effect of simvastatin versus placebo on aneurysmal subarachnoid hemorrhage were included. Two investigators independently searched articles, extracted data, and assessed the quality of included studies. The primary outcomes were delayed ischaemic deficit and delayed cerebral infarction. Meta-analysis was performed using the random-effect model. Six RCTs involving 1053 patients were included in the meta-analysis. Overall, compared with control intervention, simvastatin intervention had no influence on delayed ischaemic deficit (RR=0.99; 95% CI=0.78 to 1.27; P=0.96), delayed cerebral infarction (RR=1.17; 95% CI=0.60 to 2.29; P=0.65), mRS≤2 (RR=0.97; 95% CI=0.87 to 1.09; P=0.61), vasospasm (RR=0.79; 95% CI=0.49 to 1.29; P=0.35), ICU stay (Std. mean difference=0.04; 95% CI=-0.54 to 0.63; P=0.88), hospital stay (Std. mean difference=0.01; 95% CI=-0.13 to 0.14; P=0.90) and mortality (RR=0.71; 95% CI=0.25 to 2.05; P=0.53) after aneurysmal subarachnoid hemorrhage. Compared to control intervention, simvastatin intervention was found to have no influence on delayed ischaemic deficit, delayed cerebral infarction, mRS≤2, vasospasm, ICU stay, hospital stay, and mortality in patients with acute aneurysmal subarachnoid hemorrhage.


Primary temporary AO plate reconstruction of the mandible.
This study assessed the incidence of complications and revisions after primary temporary AO plate reconstructions of the mandible performed from 1971 through 1996. In a retrospective record review, the data of 51 patients undergoing primary temporary AO plate reconstructions after composite mandibular resection were analyzed according to age, gender, date of reconstruction, anatomic location of reconstruction, use of additional radiotherapy and/or flap surgery, and incidence of associated complications and revisions. Associated complications and revisions accompanying primary temporary AO plate reconstructions were evaluated through use of the incidence rates of associated complications (IACs) and revisions (IARs), which are reported as the numbers of associated complications and revisions, respectively, per 100 patients. Of the entire sample, the highest incidence rate of associated complications was associated with infection (33.5); this was followed by the rates for plate exposure (27.9) and plate fracture (10.7). With an overall incidence rate of 38.3 revisions per 100 patients, the corresponding site-related incidence rates of associated revisions were found to be 55.0 for reconstructions of the anterior mandible crossing the midline, 37.1 for those of the body segment of the mandible, and 31.1 for reconstructions involving the ramus and/or condyle area of the mandible. Revision rates were observed to be significantly higher in irradiated (53.6 vs 31.5) and non-flap-added reconstructions (43.2 vs 24.8). Analysis of treatment group-related incidences revealed irradiated non-flap-added reconstructions to be associated with the highest failure rates, whereas additional flap surgery resulted in a significant reduction in complications (50.0 vs 108.0) and revisions (20.0 vs 65.7). This study showed primary temporary AO plate reconstructions after composite mandibular resection to be associated with a high rate of complications and revisions. The results emphasize the need to relate outcome measures to site-related and treatment-related parameters.


High-resolution genotyping of 58 STRs in 635 Northern Han Chinese with MiSeq FGx ® Forensic Genomics System.
Sequence polymorphisms were characterized at 27 autosomal STRs (A-STRs), 7 X chromosomal STRs (X-STRs), and 24 Y chromosomal STRs (Y-STRs) in 635 Northern Han Chinese with the ForenSeq DNA Signature Prep Kit on the MiSeq FGx Forensic Genomics System. Since repeat region (RR) and flanking region (FR) variation can be detected by massively parallel sequencing (MPS), the increase in the number of unique alleles and the average of gene diversity was 78.18% and 3.51% between sequence and length, respectively. A total of 74 novel RR variants were identified at 33 STRs compared with STRSeq and previous studies, and 13 FR variants (rs1770275883, rs2053373277, rs2082557941, rs1925525766, rs1926380862, rs1569322793, rs2051848492, rs2051848696, rs2016239814, rs2053269960, rs2044518192, rs2044536444, and rs2089968964) were first submitted to dbSNP. Also, 99.94% of alleles were concordant between the ForenSeq DNA Signature Prep Kit and commercial CE kits. Discordance resulted from the low performance at D22S1045 and occasionally at DYS392, flanking region deletions at D7S820 and DXS10074, and the strict alignment algorithm at DXS7132. Null alleles at DYS505 and DYS448 and multialleles at DYS387S1a/b, DYS385a/b, DYS448, DYS505, DXS7132, and HPRTB were validated with other MPS and CE kits. Thus, a high-resolution sequence-based (SB) and length-based (LB) allele frequencies dataset from Northern Han Chinese has been established already. As expected, forensic parameters increased significantly on combined power of discrimination (PD) and combined power of exclusion (PE) at A-STRs, mildly on combined PD and combined mean exclusion chance (MEC) at X-STRs, and barely on discrimination capacity (DC) at Y-STRs. Additionally, MiSeq FGx quality metrics and MPS performance were evaluated in this study, which presented the high-quality of the dataset at 20 consecutive runs, such as ≥ 60% bases with a quality score of 20 or higher (%≥ Q20), > 60% of effective reads, > 2000 × of depth of coverage (DoC), ≥ 60% of allele coverage ratio (ACR) or heterozygote balance, ≥ 70% of inter-locus balance, and ≤ 0.4 of the absolute value of observed minus expected heterozygosity (|Hexp - Hobs|). In conclusion, MiSeq FGx can help us generate a high-resolution and high-quality dataset for human identification and population genetic studies.


The specificity of the lymphocyte transformation test in a patient with hypersensitivity reactions to pyrazolone compounds. A 10-week follow-up study before and after rechallenge.
To evaluate the specificity of the lymphocyte transformation test (LTT) in the diagnosis of drug allergy we studied over 71 days an atopic woman with a past history of frequent adverse reactions to pyrazolone drugs. Rechallenge with the incriminated substances aminophenazone (aminopyrine) and propyphenazone was carried out on Days 11 and 31 respectively. An immediate type of hypersensitivity reaction was seen after 100 mg aminophenazone, while 100 mg of propyphanozone led to a serum sickness-like syndrome. We found two specifically sensitized lymphocyte populations using either the pure substance or sera containing metabolite in cell cultures. Stimulatory responses with indices ranging between 3 and 6 were seen 3-4 days after exposure, and the tests remained positive in both instances for 3-4 weeks. Specific sensitization was proven by positive skin tests and by a small but distinct lymphocyte proliferative response before challenge. Several lymphocyte function tests were performed over a period of 53 days and revealed a large fall in pokeweed mitogen-induced immunoglobulin synthesis and an increase in suppressor cell activity after rechallenge with aminophenazone. We conclude that the proliferative response observed in the presence of the offending drug is due to the activation of T memory cells and therefore highly suggestive of a true allergic reaction.


Mitral annulus calcification and sudden death.
Mitral valve annulus calcification is a degenerative cardiac condition often found at autopsy in the elderly. While usually considered incidental to the cause of death, we report two cases where mitral valve annulus calcification with valve stenosis was associated with sudden death. Case 1: a 61-year-old female who had underlying atherosclerosis and hypertension collapsed at home. At autopsy there was marked mitral valve annulus calcification with valve stenosis and cardiomegaly. Case 2: a previously well 74-year-old female collapsed in a toilet. At autopsy there was marked calcification of the mitral valve annulus with valve stenosis. In both cases death was attributed to the effects of the calcified mitral valve annulus. Although such calcification may be unrelated to the terminal lethal mechanism, the association with left atrial enlargement, atrial fibrillation, mitral regurgitation, mitral stenosis, bacterial endocarditis, ischaemic and thromboembolic stroke, myocardial infarction, and arrhythmias, means that it should not be overlooked in the differential diagnosis in cases of sudden and unexpected death.


Quantifying and localizing actin-free barbed ends in neutrophils.
We describe here a permeablization method that retains coupling between N-formylmethionyl-leucyl-phenylalanine (fMLP) receptor stimulation and barbed-end actin nucleation in neutrophils. Using fluorescently-tagged actin monomers, we are able to quantify and localize actin-free barbed ends generated downstream of chemoattractant receptors. Partial permeabilization of the neutrophils with the mild detergent n-octyl-beta-glucopyranoside maintains signaling from membrane receptor to the actin cytoskeleton while allowing for the introduction of inhibitors and activators of signal transduction pathways implicated in regulating actin cytoskeleton dynamics. This is a useful assay for studying signal transduction to the actin cytoskeleton in neutrophils.


Invasive Brain Machine Interface System.
Because of high spatial-temporal resolution of neural signals obtained by invasive recording, the invasive brain-machine interfaces (BMI) have achieved great progress in the past two decades. With success in animal research, BMI technology is transferring to clinical trials for helping paralyzed people to restore their lost motor functions. This chapter gives a brief review of BMI development from animal experiments to human clinical studies in the following aspects: (1) BMIs based on rodent animals; (2) BMI based on non-human primates; and (3) pilot BMIs studies in clinical trials. In the end, the chapter concludes with a summary of potential opportunities and future challenges in BMI technology.


Cellasys #8: A Microphysiometric Assay for Real-Time Cell Analysis Within 24 Hours.
Traditional biochemical assays present a vital toolbox to identify the effects of a test substance on cells. However, current assays are single-point measurements, only reveal one parameter at a time, and introduce potential interferences with labels and fluorescent lights. We have addressed these limitations by introducing the cellasys #8 test which is a microphysiometric assay for real-time cell analysis. Within 24 h, the cellasys #8 test is able to identify not only the effect of a test substance but also measure recovery effects. Due to the multi-parametric read-out, the test provides insights into metabolic as well as morphological changes in real-time. The following protocol provides a detailed introduction to the materials as well as a step-by-step description to support scientists with protocol adoption. The automated and standardized assay opens up manifold new application areas for scientists to study biological mechanisms, develop new therapeutic approaches, and validate serum-free media formulation.


[Painful tibial swelling in a young patient. Findings of radiology and radionuclide scanning. Anatomopathological correlation].
We report on the case of a young man with a longstanding tumor in the left proximal tibia, in whom we had the opportunity to correlate both radiologic (plain films plus MRI) and bone scintigraphic findings with those of biopsy and clinical follow-up. The final diagnosis was bone fibrous dysplasia. The correlation between noninvasive diagnostic findings is discussed, and the convenience of using such test combination is also stressed in order to endorse a noninvasive approach to such patients, given the low malignant potential, the exception being those patients in whom there is a definite fractures risk.


[Clinical value of studying strength-duration curves using intramuscular (needle) electrodes in patients with lumbosacral radicular syndromes].
The paper deals with the results of a force-duration curve comparative study which was accomplished-in 20 patients with the aid of usual skin and acupunctural point electrodes. The spicular needle introduced painlessly into the muscles studied, served as an active electrode connected by a fine cord to a clamp of the electric pulser. In order to determine the force-duration curve, 10 rectangular impulses of a different length at a constant frequency of 1 hz were used. The results demonstrated that the determination of the force-duration curve with the aid of acupuncture is a more sensitive and precise method of study inasmuch as it permits to receive genuine values of the current in the test area.


Three-dimensional morphometric comparison of normal and apoptotic endothelial cells based on laser scanning confocal microscopy observation.
Three-dimensional (3D) morphometric analysis of cellular and subcellular structures provides an effective method for spatial cell biology. Here, 3D cellular and nuclear morphologies are reconstructed to quantify and compare morphometric differences between normal and apoptotic endothelial cells. Human umbilical vein endothelial cells (HUVECs) are treated with 60 μM H2 O2 to get apoptotic cell model and then a series of sectional images are acquired from laser scanning confocal microscopy. The 3D cell model containing plasma membrane and cell nucleus is reconstructed and fused utilizing three sequential softwares or packages (Mimics, Geomagic, and VTK). The results reveal that H2 O2 can induce apoptosis effectively by regulating the activity of apoptosis-related biomolecules, including pro-apoptotic factors p53 and Bax, and anti-apoptotic factor Bcl-2. Compared with the normal HUVECs, the apoptotic cells exhibit significant 3D morphometric parameters (height, volume and nucleus-to-cytoplasm ratio) variation. The present research provides a new perspective on comparative quantitative analysis associated with cell apoptosis and points to the value of LSCM as an objective tool for 3D cell reconstruction.


Irisin attenuates sepsis-induced cardiac dysfunction by attenuating inflammation-induced pyroptosis through a mitochondrial ubiquitin ligase-dependent mechanism.
Sepsis-induced cardiac dysfunction is a leading cause of mortality in intensive care units. However, the molecular mechanisms underlying septic cardiomyopathy remain elusive. Irisin is a cleaved product of fibronectin type III domain-containing protein 5 (FNDC5) that protects the heart from ischemia/reperfusion injury through upregulation of mitochondrial ubiquitin ligase (MITOL). Gasdermin D (GSDMD)-dependent pyroptosis plays a pivotal role in septic cardiomyopathy by regulating mitochondrial homeostasis. However, whether irisin can regulate MITOL to inhibit GSDMD-dependent pyroptosis in septic cardiomyopathy is yet to be investigated. Thus, this study was designed to explore the role of irisin in septic cardiomyopathy and its underlying molecular mechanisms. Our results demonstrate that irisin improves cardiac function against sepsis-induced cardiac dysfunction by reducing cardiac inflammation and myocardial pyroptosis. Using MITOL siRNA in vitro, the results revealed that the protective role of irisin against lipopolysaccharide (LPS)-induced cell injury was mediated by MITOL activation and the resulting inhibition of GSDMD-dependent pyroptosis. Moreover, irisin alleviated LPS-induced H9c2 cell injury by suppressing IL-1β expression and reducing serum LDH and CK-MB concentrations in a MITOL/GSDMD-dependent manner. Collectively, our data suggest that irisin treatment ameliorates cardiac dysfunction in septic cardiomyopathy by activating MITOL and inhibiting GSDMD-dependent pyroptosis. These findings highlight the clinical relevance and therapeutic potential of irisin and MITOL for the management of sepsis-induced cardiac dysfunction.


Vortioxetine Effects on Sleep Architecture in Treatment of Comorbid Depression in Adolescent Patient with Narcolepsy and Cataplexy: Case Report.
This report presents a rare case of adolescent patient treated by novel antidepressant vortioxetine for depressive disorder comorbid to narcolepsy type 1 (NT1) and newly diagnosed REM behavior disorder (RBD) and describes the overall clinical improvement of the conditions. Additionally, we discuss effect of vortioxetine on sleep architecture by evaluating objective polysomnographic studies before and on the treatment. We propose a possible efficacy of this multimodal serotoninergic agent in treatment of RBD associated with NT1.


Peak physiologic responses to arm and leg ergometry in male and female patients with airflow obstruction.
To investigate differences in work capacity for the arms and legs in patients with moderate-to-severe COPD. Cross-sectional investigation. One hundred twenty-four patients (90 men and 34 women) aged 45 to 81 years with moderate-to-very severe COPD. FEV(1) ranged from 0.70 to 2.79 L/min (FVC, 1.73 to 5.77 L; FEV(1)/FVC, 24 to 69%). All patients were in stable condition at the time of testing and receiving a stable drug regime. Each patient completed a demographic and medical history questionnaire, pulmonary function studies (spirometry, lung volumes, and diffusion capacity), peak exercise ergometry with gas exchange for the arms and legs; they also rated their subjective assessment of perceived dyspnea and extremity fatigue using Borg scores during exercise. Patients were of comparable age, with men taller and heavier than women. Smoking history was significantly less for women (47.9 pack-years vs 66.6 pack-years for men) even though each group presented with equivalent age (p > 0.05). Women were less obstructed than men, with FEV(1)/FVC (mean +/- SD) of 46.5 +/- 10.9% vs 40.2 +/- 9.3%, respectively. Ventilatory limitation during exercise was noted for all patients studied. Peak work capacity was greater for men, and leg peak responses were greater than arm values for each gender. As airway obstruction increased, work capacity became more limited. Peak arm work achieved was 38.9 +/- 19.6 W, oxygen uptake (VO(2)) was 903.9 +/- 263.5 mL/min, and minute ventilation (VE) was 33.7 +/- 9.5 L. Peak leg work value was 62.9 +/- 24.8 W, VO(2) was 1,091.4 +/- 321.5 mL/min, and VE was 39.3 +/- 12.0 L. Hence, arm values were 62%, 83%, and 85% of the measured leg values, respectively. Dyspnea and extremity effort scores were similar for men and women, and for arms and legs. Regression analysis was used to derive prediction equations for arm work from measured leg ergometry testing. For watts of work, a three-variable model emerged explaining 66% of the variance; VO(2) yielded a four-variable model with 80% of the variance explained; and VE yielded a three-variable model explaining 72% of the variance. Arm work is reduced by 38% that of the legs, while more modest reductions are noted for VO(2) and VE, suggesting greater mechanical efficiency for leg work as compared to arm work. These data also suggest greater metabolic demand for respiratory muscles and arm ergometry. Dyspnea and extremity Borg scores were equivalent for each modality and level of airway obstruction studied, suggesting that perception plays an important role in limiting exercise, and that a threshold for termination of exercise may exist. Further, peak leg ergometry results can be used with pulmonary function indexes to predict peak arm workload in watts, VO(2), and VE. These data may be used to assist the clinician in prescribing rehabilitation or estimating arm exercise ability when arm testing is unavailable.


Effect of methionine and nitrous oxide on homocysteine export and remethylation in fibroblasts from cystathionine synthase-deficient, cb1G, and cb1E patients.
We investigated the nitrous oxide-induced inactivation of methionine synthase and the concurrent homocysteine (Hcy) export in mutant fibroblasts with defects in the homocysteine catabolizing enzyme, cystathionine beta-synthase, or in methionine synthase, which carries out homocysteine remethylation. The fibroblasts were incubated in various concentrations of methionine to create conditions favoring methionine conservation or catabolism. In cystathionine beta-synthase-deficient cells, high medium methionine partly protected the enzyme against inactivation, as previously found in normal fibroblasts. The Hcy export rate at low methionine levels was low (0.2-0.6 nmol/h/10(6) cells), and increased 2-3-fold at high methionine levels. Nitrous oxide enhanced Hcy export rate at low methionine, so that in the presence of nitrous oxide, the Hcy export became less dependent of methionine. In cb1G cells, the enzyme inactivation was moderate and independent of medium methionine. The Hcy export rate was intermediate (0.5-0.8 nmol/h/10(6) cells) at low methionine levels, and increased moderately (< 2-fold) at high methionine levels or following nitrous oxide exposure. In cb1E mutants, the enzyme activity was not affected by nitrous oxide, and the Hcy export was high (0.8-1.6 nmol/h/10(6) cells) and independent of methionine and nitrous oxide. These data suggest that Hcy remethylation and cystathionine beta-synthase activity are major determinants of Hcy export at low and high methionine, respectively. The low susceptibility of methionine synthase to nitrous oxide in the presence of high methionine or in cb1G or cb1E mutants is probably related to low catalytic turnover.


Tris-heteroleptic ruthenium(II) polypyridyl complexes: Synthesis, structural characterization, photophysical, electrochemistry and biological properties.
Three water-soluble tris-heteroleptic ruthenium(II) polypyridyl complexes [Ru(bpy)(phen)(bpg)]2+ (1), [Ru(bpy)(dppz)(bpg)]2+ (2), and [Ru(phen)(dppz)(bpg)]2+ (3) (where bpy = 2,2'-bipyridine, phen = 1,10-phenanthroline, dppz = dipyrido[3,2-a:2',3'-c] phenazine, bpg = 4b,5,7,7a-tetrahydro-4b,7a-epiminomethanoimino-6H-imidazo[4,5-f] [1,10] phenanthroline-6,13-dione) have been synthesized and characterized. Molecular structures of complexes 1 and 3 are confirmed by single crystal X-ray structure determination. Interaction of complexes 1-3 with DNA is explored by various spectroscopic techniques. The complexes 1-3 show solvent dependent photophysical properties. Complexes 2 and 3 show extensive "molecular light switch" effect for DNA. The complexes 1-3 are low toxic towards HeLa (human cervical cancer) and HL-60 (human promyelocytic leukemia) cell lines. Further, the cellular uptake of complexes 2 and 3 by cells shows that complexes mainly localised on the nucleus of the cells.


Scaling Up Public Health Interventions: Engaging Partners Across Multiple Levels.
Advancing the science of intervention scale-up is essential to increasing the impact of effective interventions at the regional and national levels. In contrast with work in high-income countries (HICs), where scale-up research has been limited, researchers in low- and middle-income countries (LMICs) have conducted numerous studies on the regional and national scale-up of interventions. In this article, we review the state of the science on intervention scale-up in both HICs and LMICs. We provide an introduction to the elements of scale-up followed by a description of the scale-up process, with an illustrative case study from our own research. We then present findings from a scoping review comparing scale-up studies in LMIC and HIC settings. We conclude with lessons learned and recommendations for improving scale-up research.


Doctor knows best? Provider bias in the context of contraceptive counseling in the United States.
The study examined how clinicians described their patients in relation to their practices of contraceptive counseling. This qualitative study involved individual interviews with 15 clinicians working in obstetrics and gynecology in South Carolina about their approaches to contraceptive counseling. We analyzed the data using a combination of deductive and inductive approaches. Clinicians attributed challenges of working with diverse patient populations to patients' race/ethnicity, socioeconomic status, and age. Clinicians often interpreted patient concerns about or refusal to use effective contraceptive methods as a problem with patients themselves. When clinicians described patients in disparaging ways, they often focused on adolescent patients. Bias informed by structural inequalities and power relations influences how clinicians perceive their patients and approach counseling them about contraception. Such practices may limit patients' informed decision-making and autonomy regarding initiating or continuing contraceptive use. Greater attention to redressing structural inequalities and power relations that inform provider bias in the context of contraceptive counseling is needed to ensure patients receive person-centered healthcare free from prejudice and discrimination. Fostering structural competency among clinicians may improve provider-patient interactions and support patients' reproductive autonomy.


Fertilizer stabilizers reduce nitrous oxide emissions from agricultural soil by targeting microbial nitrogen transformations.
Nitrous oxide (N2O) is a pollutant released from agriculture soils following N fertilizer application. N stabilizers, such as N-(n-butyl) thiophosphoric triamide (NBPT) and 3,4-dimethylpyrazole phosphate (DMPP) could mitigate these N2O emissions when applied with fertilizer. Here, field experiments were conducted to investigate the microbial mechanisms by which NBPT and DMPP mitigate N2O emissions following urea application. We determined dynamic N2O emissions and inorganic N concentrations for two wheat seasons and combined this with metagenomic sequencing. Application of NBPT, DMPP, and both NBPT and DMPP together with urea decreased mean N2O accumulative emissions by 77.8, 91.4 and 90.7%, respectively, compared with urea application alone, mainly via repressing the increase in NO2- concentration after N fertilization. Sequencing results indicated that urea application enriched microorganisms that were positively correlated with N2O production, whereas N stabilizers enriched microorganisms that were negatively correlated with N2O production. Furthermore, compared to urea application alone, NBPT with urea reduced the abundances of genes related to denitrification, including napA/nasA, nirS/nirK, and norBC, resulting in a higher soil NO3- pool. Conversely, DMPP application, either alone or together with NBPT, decreased the abundance of genes involved in ammonia oxidation and denitrification, including amoCAB, hao, napA/nasA, nirS/nirK, and norBC, and maintained a greater soil NH4+ pool. Both N stabilizers resulted in similar abundances of nirABD-which is related to NO2- reducers-as when no N fertilizer was applied, which could prevent NO2- accumulation, consequently mitigating N2O emissions. These findings suggest that the high effectiveness of N stabilizers on mitigating N2O emissions could be attributed to changes to soil microbial communities and N-cycling functional genes to control the by-product or intermediate products of microbial N-cycling processes in agricultural soils.


Changes in health-related quality of life are associated with patient satisfaction following total hip replacement: an analysis of 69,083 patients in the Swedish Hip Arthroplasty Register.
Background and purpose - Total hip replacement (THR) aims mainly to improve quality of life via restoration of hip function and provision of pain relief. This study sought to assess whether improvements in quality of life between the preoperative and 1-year postoperative period were associated with patient satisfactionPatients and methods - Data were extracted for 69,083 THR operations with complete data reported to the Swedish Hip Arthroplasty Register (SHAR) between 2008 and 2015. Health-related quality of life and patient satisfaction were captured using the Euro-Qol-5D (EQ-5D) and visual analogue scale (VAS), respectively. Multivariable analysis was performed to assess associations between the changes in pre- and postoperative EQ5D and patient satisfaction.Results - In patients reporting severe or moderate problems with mobility preoperatively, improvement to no problems was associated with numerically higher patient satisfaction (coefficient -18 [95% CI -22 to -14] and -18 [-18 to -17]). Improvement in the self-care dimension from severe or moderate problems to no problems was associated with numerically higher patient satisfaction (-15 [-16 to -14] and -13 [-15 to -11]). Improvement from severe problems with the ability to perform usual activities to no problems was associated with numerically higher patient satisfaction (-18 [-19 to -17]). This association was also found for improvement in pain/discomfort and anxiety/depression (-16 [-17 to -15] and -15 [-16 to -14]).Interpretation - Our results indicate that satisfaction with the operated hip is a valid patient-reported outcome reflecting the changes in different EQ-5D dimensions and should be included in the follow-up of patients after THR surgery.


E-Commerce Policy and International Business.
The rise of digitization and information and communication technologies (ICT) is playing a vital role in facilitating global trade and business activities and in overcoming cross-border transaction costs. In so doing, it offers firms significant benefits and opportunities to compete on a global scale, as witnessed during the ongoing COVID-19 pandemic. The growth and widespread diffusion of internet-enabled technologies and platforms have created numerous opportunities for firms to provide products and services across both developed and developing markets. Yet, limited research has been conducted in the international business domain to explore the rise of ecommerce and its implications for international business scholarship. In this focused issue, we present an examination of the role played by e-commerce in international business, paying particular attention to the policy aspect of e-commerce and issuing a call for a greater integration of e-commerce policy in international business research.


Tracing all patients who received insured dialysis treatment in Japan and the present situation of their number of deaths.
The survival rate of chronic dialysis patients in Japan remains the highest worldwide, so there is value in presenting Japan's situation internationally. We examined whether aggregate figures on dialysis patients in the National Database of Health Insurance Claims and Special Health Checkups of Japan (NDB), which contains data on insured procedures of approximately 100 million Japanese residents, complement corresponding figures in the Japanese Society for Dialysis Therapy Renal Data Registry (JRDR). Subjects were patients with medical fee points for dialysis recorded in the NDB during 2014-2018. We analyzed annual numbers of dialysis cases, newly initiated dialysis cases- and deaths. Compared with the JRDR, the NDB had about 6-7% fewer dialysis cases but a similar number of newly initiated dialysis cases. In the NDB, the number of deaths was about 6-10% lower, and the number of hemodialysis cases was lower, while that of peritoneal dialysis cases was higher. The cumulative survival rate at dialysis initiation was approximately 6 percentage points lower in the NDB than in the JRDR, indicating that some patients die at dialysis initiation. Cumulative survival rate by age group was roughly the same between the NDB and JRDR in both sexes. The use of the NDB enabled us to aggregate data of dialysis patients. With the definition of dialysis patients used in this study, analyses of concomitant medications, comorbidities, surgeries, and therapies will become possible, which will be useful in many future studies.


Bismuth release from endodontic materials: in vivo analysis using Wistar rats.
Calcium silicate-based materials are used to block the communication between the root canal and the periodontal ligament space. This brings the materials into contact with tissues and the potential for local and systemic elemental release and movement. The aim of the study was to evaluate the elemental release of bismuth from ProRoot MTA in contact with connective tissues after 30 and 180 days as well as any accumulation in peripheral organs using an animal model. Tricalcium silicate and hydroxyapatite containing 20% bismuth oxide (HAp-Bi) were used as controls. The null hypothesis was that bismuth migrates from tricalcium silicate-based materials when associated with silicon. The materials were examined using scanning electron microscopy, energy dispersive spectroscopy (SEM/EDS) and X-ray diffraction prior to implantation as well as using SEM/EDS, micro X-ray fluorescence and Raman spectroscopy after implantation to assess elemental presence in surrounding tissues. Histological analysis was used to evaluate the changes in tissue architecture and inductively coupled plasma mass spectrometry (ICP-MS) was used to investigate the elemental deposition. For the systemic investigation, routine blood analysis was performed and organs were obtained to evaluate the presence of bismuth and silicon using ICP-MS after acid digestion. In the histological analysis of the implantation sites, macrophages and multinucleated giant cells could be observed after 30 days which after 180 days became a chronic infiltrate; although, no major differences were identified in red and white blood cell analyses and biochemical tests. Implantation altered the materials as observed in the Raman analysis and bismuth was detected both locally and within kidney samples after both periods of analysis, indicating the potential for accumulation of bismuth in this organ. Smaller amounts of bismuth than observed in the kidney were also detected in blood, liver and brain for the ProRoot MTA and HAp-Bi after 180 days. Bismuth was released from the ProRoot MTA locally and was detected systemically and in samples without silicon; thus, the null hypothesis was rejected. The bismuth release demonstrated that this element accumulated both locally and systemically, mainly in the kidneys in comparison with brain and liver regardless of the material base.


Application of feeding guiders to improve the powder distribution in the two scales of roller compactors.
Roller compaction is a continuous dry granulation process, where the powder is compressed between two counter-rotating rollers and compacted into ribbons. The quality and homogeneity of the granulate is determined by the uniformity and porosity of the ribbon, which depends on the feeding process of the primary powder to the rollers, the flow properties of the primary powder and process parameters such as roller forces. Previous work was conducted to improve the powder flow and distribution in the feeding zone by developing new feeding guiders, which are located in the feeding zone close to the rollers on the lab-scale roller compactor Alexanderwerk WP120 Pharma (Yu et al., 2018). These new feeding guiders were used to reduce the amount of powder that is delivered to the centre of the rollers and increase the amount of powder that is delivered to the sides of the rollers, in comparison to the original feeding guiders. This modified concept using new feeding guiders has been applied to the large-scale roller compactor Alexanderwerk WP200 Pharma in the present work. In order to evaluate the homogeneity of the ribbon properties across the ribbon width, the temperature profile and porosity distribution across the ribbon width were measured. The new feeding guiders resulted in ribbons being produced with a more uniform temperature profile and porosity distribution across the ribbon width when using the small and large scale roller compactors at different process parameters.


Chronic pre-treatment with nicotine enhances nicotine-evoked striatal dopamine release and alpha6 and beta3 nicotinic acetylcholine receptor subunit mRNA in the substantia nigra pars compacta of the rat.
Whilst local intrastriatal infusion of nicotine consistently elicits striatal dopamine release, systemic administration often fails to do so. Since chronic nicotine administration is known to result in desensitisation-induced upregulation of nicotinic acetylcholine receptors (nAChRs), the present study investigated whether chronic pre-treatment could enhance the response to systemic nicotine and, if so, whether increases in specific nAChR subunit mRNA levels in the substantia nigra pars compacta (SNc) may underlie this effect. In vivo microdialysis studies in male Sprague-Dawley rats revealed that following 4 days pre-treatment with nicotine (0.8 mg kg(-1)s.c.), local intrastriatal nicotine infusion (3 mM) elicited significantly higher dopamine efflux compared to vehicle pre-treated controls (peak release: 1273 +/- 199% basal versus 731 +/- 113% basal), whereas systemic nicotine challenge (0.8 mg kg(-1)s.c.) elicited no response. In contrast, following 8 days pre-treatment with nicotine (0.8 mg kg(-1)s.c.), systemic nicotine challenge (0.8 mg kg(-1)s.c.) now produced significantly higher dopamine efflux than that of vehicle pre-treated controls (147 +/- 30% basal versus 91 +/- 5% basal). Eight days pre-treatment with nicotine also significantly elevated the levels of alpha6 (approximately 55%) and beta3 (approximately 43%) nAChR subunit mRNA in the SNc, suggesting that up-regulation of these nAChR subunit genes in the nigrostriatal tract may contribute to the enhanced nicotine-evoked striatal dopamine release.


In vitro proliferation of aortic smooth muscle cells from spontaneously hypertensive and normotensive rats.
The characteristics and proliferation of aortic smooth muscle cells (SMC) from spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto (WKY) rats were studied in culture. Smooth muscle cells were isolated from the tunica media of the thoracic aorta by an explant method. Immunofluorescence microscopy showed that 93-95 per cent of cells were positively labelled with antibodies raised against smooth muscle actin, indicating that these were smooth muscle cells. The proliferative activity was compared between aortic smooth muscle cells from hypertensive and normotensive rats in culture by thymidine incorporation and cell number determinations. The results demonstrate that aortic smooth muscle cells from hypertensive rats grew faster than those from normotensive rats in culture. The increased proliferative activity of cultured aortic smooth muscle cells from hypertensive rats was detectable even when they were cultured in a chemically defined serum-free medium. These data have shown that an increased proliferative activity of aortic smooth muscle cells from hypertensive rats can occur in culture conditions without the influence of arterial pressure or other stimuli as in intact animals. The mechanisms underlying the accelerated proliferative activity of aortic smooth muscle cells from genetically hypertensive rats in vitro remain to be determined.