Pacemaker architecture: a pacemaker with an attached computer or a computer with an attached pacemaker.
Microprocessors have a major impact on cardiac pacemaker technology. The development of adaptive systems capable of responding to physiologic variables will ultimately improve the care of the individual patient. However, in order to minimize power drain, it will be necessary to incorporate certain functions into the hardware outside the microprocessor. In addition, specifically designed program languages may indeed become necessary.


Defined megadalton hyaluronan polymer standards.
The utility of polymer standards for the calibration of average molecular mass estimates often is limited by the polydispersity--the breadth of the size distribution--of the standard. Here monodisperse synthetic hyaluronan (or hyaluronic acid [HA]) complexes in the approximately 1- to 8-megadalton (MDa) range were prepared in two steps. First, synchronized stoichiometrically controlled in vitro reactions yielded linear narrow size distribution biotinylated HA chains. Second, streptavidin protein was added at substoichiometric levels to prepare a series of complexes with one, two, three, or four HA chains per streptavidin molecule. The dendritic-like molecules approximate the mobility of natural linear HA chains on agarose gels, making the complexes useful as defined size standards for high-molecular weight HA preparations.


Combining PD-L1 blockade with immunogenic cell death induced by AIE photosensitizer to improve antitumor immunity.
Immunogenic cell death (ICD) is considered an effective death mode to trigger immune response. However, the currently available efficient ICD inducers are quite limited. Endoplasmic reticulum (ER) stress is known as the precursor of ICD, which can be directly triggered by reactive oxygen species in situ. Herein, a novel photosensitizer (α-Th-TPA-PIO) based on phosphindole oxide, featuring aggregation-induced emission (AIE) is designed and prepared, which possesses good ability of hydroxyl radicals (HO•) generation. Besides, α-Th-TPA-PIO can selectively accumulate in ER and trigger ER stress under white light irradiation, further leading to effective ICD. Combining with anti-programmed death-ligand 1 (anti-PD-L1), the synergistic effect of photodynamic therapy (PDT) and immune checkpoint blockade can achieve a significantly enhanced inhibition effect on the growth of tumors and simultaneously provoke a systemic antitumor immune response. Notably, by adopting this therapeutic strategy to bilateral and metastatic tumor models, the growth of both primary and distant subcutaneous tumors can be successfully suppressed, and metastatic tumor can also be inhibited to some degree. Taken together, this work not only provides a novel ICD photoinducer based on PDT, but also brings about a useful immunomodulatory strategy to realize superior antitumor effect.


Modulation of heat shock protein response in SH-SY5Y by mobile phone microwaves.
To investigate putative biological damage caused by GSM mobile phone frequencies by assessing electromagnetic fields during mobile phone working. Neuron-like cells, obtained by retinoic-acid-induced differentiation of human neuroblastoma SH-SY5Y cells, were exposed for 2 h and 4 h to microwaves at 1800 MHz frequency bands. Cell stress response was evaluated by MTT assay as well as changes in the heat shock protein expression (Hsp20, Hsp27 and Hsp70) and caspase-3 activity levels, as biomarkers of apoptotic pathway. Under our experimental conditions, neither cell viability nor Hsp27 expression nor caspase-3 activity was significantly changed. Interestingly, a significant decrease in Hsp20 expression was observed at both times of exposure, whereas Hsp70 levels were significantly increased only after 4 h exposure. The modulation of the expression of Hsps in neuronal cells can be an early response to radiofrequency microwaves.


Predictors of outcomes in children with Central Nervous System tuberculosis.
Central Nervous system tuberculosis (CNS-Tb) is the most lethal form of extra-pulmonary tuberculosis in children. The lack of markers of outcome provides little information on the efficacy of the current treatment protocols for CNS-Tb and thus results in a higher mortality rate than other extrapulmonary manifestations of tuberculosis. This study aims to identify significant factors that will reliably predict the outcomes at discharge in children admitted with CNS-Tb. This is a prospective observational study in children with neurotuberculosis admitted at a tertiary care hospital. Clinical presentations at the time of admission were studied. Outcomes at the end of in-patient care (completely cured, survival with some/severe disability or death) were correlated with clinical, laboratory, microbiological, and radiological parameters. Univariate and multivariate analyses were applied to study the parameters and a p-value ≤ 0.05 with a confidence interval (CI) of 95% was considered as statistically significant. The study included 100 children between 4 months and 12 years of age with a mean of 5.84 (±3.5) years. At discharge, 55% of children recovered completely, 20% had some or severe disability and 25% died. On multivariate analysis, high CSF protein (p = 0.050) and drug resistance (p = 0.034) were highly associated with fatality. Meningeal enhancements with basal exudates (p = 0.021) and CSF lymphocyte count >90% were highly associated with survival with disability. Stage I disease at presentation (p < 0.0001) was the only variable associated with complete recovery. Reliable prognostic markers for CNS-Tb can aid in predicting the efficacy of the current treatment and the anticipated outcome in the children with this disease. This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.


Differences in redox and kinetic properties between NAD-dependent and O2-dependent types of rat liver xanthine dehydrogenase.
Reductive titrations of a NAD-dependent type (type-D) and an O2-dependent type (type-O) of rat liver xanthine dehydrogenase showed that only the type-D enzyme formed a pronounced stable FAD semiquinone (FADH*). The FAD semiquinone was less stabilized in the presence of NAD. The Vmax value for xanthine-NAD activity of type-D enzyme was close to that for xanthine-O2 activity of type-O enzyme, while the Vmax value for xanthine-O2 activity of type-D enzyme was about one-fourth of that of type-O enzyme. The Km value for O2 of type-D enzyme was about five times as large as that of type-O enzyme. The absorbance spectrum of type-D enzyme during turnover with xanthine and O2 as substrates showed a considerable amount of FADH* formation, but that with xanthine and NAD as substrates showed only a negligible one. Low xanthine-O2 activity of type-D enzyme, as compared with that of type-O enzyme, seems to be explained by the conformational change occurring in conversion from type-O to type-D enzyme, which results in different reactivity of FAD to molecular oxygen and a higher fraction of FADH* during turnover. The binding of NAD may possibly increase the fraction of FADH2, resulting in a Vmax value of xanthine-NAD activity almost as high as that of xanthine-O2 activity of type-O enzyme.


Observational study of surgical resection in small non-functional pancreatic neuroendocrine tumors: AS SEER-based study.
The potential benefits of surgical resection for small non-functional pancreatic neuroendocrine tumors (NF-PNETs) in terms of survival remain uncertain. This study aimed to evaluate the impact of surgical treatment on patients with NF-PNETs. Using SEER data, we identified 1102 patients from 2004 to 2015 with well and moderately differentiated pancreatic neuroendocrine tumors (PNETs). The associations between continuous variables and receipt of surgery were assessed using Wilcoxon rank-sum tests. Kaplan-Meier survival curves for OS were compared using the log-rank test. We compared outcomes in patients who received surgical resection with those in patients who did not, using a univariable Cox model with inverse probability weighting according to the propensity score and propensity-score matching. Among the cohort of 1102 patients, a majority of 965 individuals (87%) underwent surgical intervention. Upon conducting univariate analysis, we observed that surgical treatment significantly prolonged patients' survival [HR = 0.41, 95% CI [0.26-0.65] P < 0.001]. However, the old [HR = 3.27, 95% CI (2.24-4.76), P 0.001], male gender [HR = 1.82, 95% CI (1.23-2.68), P = 0.003], and moderately well-differentiated factors [HR = 1.71, 95% CI (1.04-2.80), P = 0.034] were found to potentially decrease patients' survival time. In the multivariate analysis, male gender [HR = 1.73, 95% CI (1.15-2.61), P = 0.009] and the old factor [HR = 3.52, 95% CI (2.33-5.31), P < 0.001] emerged as influential predictors with higher hazard ratios. Notably, surgical treatment remained a significant factor associated with improved overall survival [HR = 0.53, 95% CI (0.33-0.84), P = 0.007]. Propensity-score matching and inverse probability weighting were employed as analytical techniques. The univariate analysis results showed favorable outcomes in the weight group [HR = 0.48, 95% CI (0.29-0.78), P = 0.003] and matched group [HR = 0.44, 95% CI (0.22-0.85), P = 0.015], respectively. Survival analysis further confirmed that surgical treatment contributed to increased overall survival (log rank, P < 0.05) in both the matching and weight groups. Patients diagnosed with small, non-functioning pancreatic neuroendocrine tumors who undergo surgical intervention exhibit improved overall survival (OS) outcomes. Therefore, surgery is strongly recommended for this patient population.


Serum testosterone levels and mortality in men with CKD stages 3-4.
Hypogonadism in men (total testosterone <350 ng/dL) is associated with higher risk of cardiovascular disease and mortality in men on dialysis therapy. We evaluated the association of hypogonadism with all-cause mortality in men with non-dialysis-dependent chronic kidney disease (CKD). Retrospective, cohort study. 2,419 men with CKD stages 3-4 (estimated glomerular filtration rate, 15-59 mL/min/1.73 m2) who had total testosterone measured for cause between January 1, 2005, and October 31, 2011, at a tertiary-care center in Cleveland, OH. Total testosterone measured using an immunoassay measurement in 3 forms: (1) categorized as low or testosterone replacement therapy versus normal, (2) continuous log testosterone, and (3) quintiles (100-226, 227-305, 306-392, 393-511, and 512-3,153 ng/dL). Factors associated with low total testosterone level and the association between low total testosterone level and all-cause mortality were evaluated using logistic regression, Cox proportional hazard models, and Kaplan-Meier survival curves. Hypogonadism was found in 1,288 of 2,419 (53%) men. In a multivariable logistic regression analysis, African American ethnicity and higher estimated glomerular filtration rate were associated with lower odds of having hypogonadism. Diabetes and higher body mass index were associated with higher odds of having hypogonadism. 357 of 2,419 (15%) patients died during a median follow-up of 2.3 years. In the multivariate Cox model, testosterone level <350 ng/dL or testosterone replacement therapy was not associated with mortality. In a multivariable model also adjusted for testosterone supplementation, higher log testosterone was associated with significantly lower mortality (HR per 1 log unit, 0.70; 95% CI, 0.55-0.89). When compared to the highest quintile, the second lowest quintile of testosterone was associated with higher mortality (HR, 1.53; 95% CI, 1.09-2.16). Single-center study, timing of testosterone testing, lack of adjustment for proteinuria, and sampling bias. Low total testosterone level may be associated with higher mortality in men with CKD stages 3-4, but more studies are needed.


Disengagement of general practitioners in cervical cancer screening.
In the absence of organized cervical cancer screening (CCS) programs, gynecologists remain principal actors in obtaining a Pap smear, followed by general practitioners (GPs). In France, with the growing scarcity of gynecologists and social inequalities in access to opportunistic screening, GPs are valuable resources for women's gynecologic follow-up. We aimed to investigate the characteristics of GPs who do not perform CCS, analyzing the effect of GPs' sex and their evolution over time. On the basis of data from three cross-sectional surveys conducted among representative samples of French GPs in 1998, 2002, and 2009 (n=5199), we constructed univariate and multivariate logistic mixed models (level 2: county, level 1: GP) with random intercept stratified on GPs' sex to investigate the characteristics of the GPs associated with no practice of CCS ever. Almost one-third of all GPs did not perform CCS ever and it increased with time. Male GPs were always more likely not to perform it (odds ratio=0.50, 95% confidence interval=0.42-0.59). The percentage of GPs not performing CCS increased more markedly among male than among female GPs, and increased more among the youngest age group. Increasingly fewer GPs engage in CCS when the growing scarcity of medical gynecologists calls for more participation. Female GPs remain significantly more active in CCS than male GPs. The participation in CCS is determined differently according to the practitioner's sex.


Macrophage-Biomimetic Nanoparticles Ameliorate Ulcerative Colitis through Reducing Inflammatory Factors Expression.
Inflammatory mediator S100A9 is dramatically elevated in ulcerative colitis and correlates with disease severity. S100A9 is a potential molecule to target for the treatment of colitis, but to date, there is no effective targeting method. The aim of this study was to develop a safe and effective nano-delivery system targeting S100A9 and to evaluate its therapeutic efficacy in ulcerative colitis mouse model. We designed an oral nano-delivery system using poly (lactic acid-glycolic acid) (PLGA)-loaded S100A9 inhibitor tasquinimod to synthesize PLGA-TAS nanoparticles. TLR4-overexpressing macrophage membranes (MMs) were used to wrap the nanoparticles to make MM-PLGA-TAS, which allowed the nanoparticles to acquire the ability to specifically enrich the colitis region. MM-PLGA-TAS was endocytosed by inflammatory phenotype RAW264.7 cells in vitro and can efficiently enrich in inflamed mouse colitis tissue in vivo. A chemically induced ulcerative colitis mouse model was used to evaluate the therapeutic effect of oral MM-PLGA-TAS. MM-PLGA-TAS significantly alleviated the symptoms of ulcerative colitis, and mechanically, MM-PLGA-TAS achieved immunomodulatory and suppressive effects by reducing S100a9 and other cytokines in the colitis region. We describe a convenient, orally targeted colitis drug delivery system that cures the disease in ulcerative colitis mice. This system substantially increases drug accumulation in inflamed colonic tissue, reduces the risk of systemic exposure, and is a promising therapeutic approach against ulcerative colitis.


Integrated microRNA‑gene analysis of coronary artery disease based on miRNA and gene expression profiles.
The present study aimed to investigate the key genes and microRNAs (miRNA/miRs) associated with coronary artery disease (CAD) progression. The gene expression profile of GSE20680 and GSE12288, and the miRNA expression profile of GSE28858 were downloaded from the gene expression omnibus database. The differentially expressed genes (DEGs) in GSE20680 and GSE12288, and the differentially expressed miRNAs in GSE28858 were screened using the limma package in R software. Common DEGs between GSE20680 and GSE12288 were selected. Functions and pathways of DEGs and miRNAs were enriched using the DAVID tool from the GO and KEGG databases. The regulatory network of miRNA and selected CAD‑associated DEGs was constructed. A total of 270 DEGs (167 upregulated and 103 downregulated) based on the GSE20680 dataset, and 2,268 DEGs (534 upregulated and 1,734 downregulated) based on the GSE12288 dataset, were screened. For the differentially expressed miRNAs, 214 were identified (102 upregulated and 112 downregulated) in CAD samples and were screened. Interferon regulatory factor 2 (IRF2) and cell death‑inducing DFFA‑like effector b (CIDEB), which are regulated by signal transducer and activator of transcription 3 and myc‑associated factor X, were identified as common DEGs for CAD. miR‑455‑5p, miR‑455‑3p and miR‑1257, which are involved in the major histocompatibility complex (MHC)protein assembly pathway and peptide antigen assembly with MHC class I protein complex pathway, may regulate various miRNAs and target genes, including pro‑opiomelancortin (POMC), toll‑like receptor 4 (TLR4), interleukin 10 (IL10), activating transcription factor 6 (ATF6) and calreticulin (CALR). The current study identified IRF2 and CIDEB as crucial genes, and miRNA‑455‑5p, miRNA‑455‑3p and miR‑1257 along with their target genes POMC, TLR4 and CALR, as miRNAs involved in CAD progression. Thus, the present study may provide a basis for future research into the progression mechanism of CAD.


The FIB-4 index is a significant prognostic factor in patients with non-B non-C hepatocellular carcinoma after curative surgery.
The aspartate aminotransferase to platelet ratio index (APRI) and fibrosis-4 (FIB-4) index were developed as a non-invasive parameter for predicting liver fibrosis. This study aimed to validate the APRI and FIB-4 indexes in patients treated with curative therapy for non-B non-C (NBNC) hepatocellular carcinoma (HCC). Accumulated database comprising 399 patients who underwent hepatectomy was reviewed retrospectively. Analyses were performed to evaluate whether the APRI and FIB-4 indexes are predictors of liver cirrhosis and/or the prognosis in patients with NBNC-HCC. Forty-seven patients with NBNC-HCC who underwent curative radiofrequency ablation therapy (RFA) in the same period were enrolled as the validation set. The APRI and FIB-4 indexes were significantly higher in the cirrhosis group than in the no-cirrhosis group (P = 0.001 and P < 0.001, respectively). A receiver operating characteristic curve analysis showed that the FIB-4 index was more accurate in predicting background liver cirrhosis than the APRI. According to a multivariate analysis, an FIB-4 index larger than 2.7 (hazard ratio 2.11 and 2.21, 95 % confidence interval 1.06-4.18 and 1.38-3.54, P = 0.033 and P = 0.001) remained significant independent predictors of overall and recurrence-free survival, respectively. The present findings showed that the FIB-4 index is a significant predictor of background liver cirrhosis and the prognosis after curative resection for NBNB-HCC.


Serum glycoproteins in the liver diseases. V. Desialylated glycoproteins in chronic hepatitis.
Serum desialylated glycoprotein level was tested for chronic hepatitic patients. The level was significantly elevated in patients with chronic aggressive hepatitis but not in chronic persistent hepatitis comparing to normal subjects. In chronic aggressive hepatitis, severe type (2B), serum desialylated glycoprotein levels were significantly enhanced but not in moderate type (2A) when compared to chronic persistent hepatitis. Sera taken serially from patients with chronic aggressive hepatitis, severe type (2B), demonstrated a slight correlation between circulating desialylated glycoprotein level and serum glutamic-pyruvic transaminase activity.


Effect of gas components on the degradation mechanism of o-dichlorobenzene by non-thermal plasma technology with single dielectric barrier discharge.
In this paper, the degradation of o-DCB under different gas-phase parameter conditions was investigated using the SDBD-NTP system. The results showed that the increase in initial and oxygen concentrations had opposite effects on the degradation of o-DCB. Among them, the increase of oxygen concentration promoted the degradation of o-DCB. Relative humidity promoted and then inhibited the degradation of o-DCB. The highest degradation efficiency of o-DCB was achieved at RH = 15%, reaching 91% at 29W. In the study of by-products, it was found that O3 and NOx were the main inorganic by-products, and that different oxygen levels and relative humidity conditions had a large effect on the production of O3 and NOx. In all of them, the concentration of O3 decreased with the increase of input power. NOx increased with increasing oxygen concentration, but the increase in relative humidity inhibited the production of NO and N2O and promoted the conversion of NO2. A study of organic by-products revealed this. In the absence of oxygen, a higher number of benzene products appeared. Whereas, with the addition of oxygen, only in the by-products under conditions where no relative humidity was introduced, benzene ring products were predominantly present in the by-products. However, when RH was added, n-hexane was found to be present in the by-products. This may be because the introduction of OH• favors the destruction of the benzene ring. Finally, the possible reaction pathways and reaction mechanisms of o-DCB under different gas-phase parameters are given. It provides a reference for future related scientific research as well as scientific problems in practical applications.


Insulin action is normalized in newly diagnosed type I diabetic patients after three months of insulin treatment.
We studied insulin action at submaximal and maximal insulin levels in seven newly diagnosed type I (insulin-dependent) diabetic patients after 2 weeks (t1/2) and after 3 months (t3) of insulin treatment, and in seven control subjects. Insulin action was determined with a sequential euglycemic (5.0 mmol/L) glucose clamp technique using insulin infusion rates of 0.5, 1.0, 2.0, and 5.0 mU.kg-1.min-1 in four periods of two hours each. The final 30 minutes of each infusion period (referred to as steady-state) were taken for the assessment of insulin action. Steady-state insulin levels were similar in the diabetic patients at t1/2 and t3, and in control subjects. During the first and second infusion periods, steady-state glucose infusion rates (SSGIR) were lower at t1/2 than at t3 (12.2 +/- 1.7 v 18.8 +/- 2.4, P less than .05, and 34.3 +/- 3.8 v 47.6 +/- 2.5 mumol.kg-1.min-1, P less than .02, respectively), and were lower at t1/2 compared to controls (12.2 +/- 1.7 v 22.4 +/- 2.3, P less than .01, and 34.3 +/- 3.8 v 47.3 +/- 2.8 mumol.kg-1.min-1, P less than .02). No differences were found during the third and fourth infusion periods between t1/2 and t3, or t1/2 and controls. When these data were used to construct dose-response curves, insulin action was decreased in the diabetic patients at t1/2 at submaximal insulin levels (shift to the right), while insulin responsiveness was unchanged. This finding may be regarded as a still-present manifestation of the metabolic derangement at the onset of the disease.(ABSTRACT TRUNCATED AT 250 WORDS)


Spectral characterization, thermochemical studies, periodic SAPT calculations and detailed quantum mechanical profiling various physico-chemical properties of 3,4-dichlorodiuron.
A set of experimental and computational techniques have been applied for the understanding of fundamental spectroscopic and reactive properties of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (diuron) compound. Experimental techniques employed in this study encompassed spectroscopic characterization via IR and Raman approaches, while optical properties were studied by measurements of UV/Vis spectra. The thermogravimetric analysis was also studied in order to analyze the stability of diuron. Aside from the determination of reactive properties, DFT calculations on isolated molecules were also used to thoroughly visualize and analyze spectroscopic properties such as IR and UV/Vis. MD simulations were used in order to understand interactions with water, while periodic DFT calculations were used in order to analyze band structure and density of states of the diuron crystal structure. Since the crystal structure of diuron is known, it was used in order to extract the relevant molecular pairs and investigate interactions between them by DFT and symmetry adapted perturbation theory approaches (SAPT).


Left atrial function index predicts poor outcome in STEMI patients treated with percutaneous coronary intervention.
The prognostic value of the left atrial function index (LAFI) in acute ST segment elevation myocardial infarction (STEMI) patients treated with percutaneous coronary intervention (PCI) is unknown. This study sought to determine whether the LAFI predicts prognosis in STEMI patients treated with PCI. Patients with newly diagnosed STEMI who were treated with PCI in Hunan Provincial People's Hospital from March 2020 to October 2020 were prospectively enrolled. All patients underwent transthoracic echocardiography at baseline and follow-up. The endpoint events included rehospitalization due to unstable angina, nonfatal myocardial infarction, rehospitalization due to heart failure and cardiovascular death. A total of 156 STEMI patients treated with PCI were studied with a median follow-up of 14 months. Forty-eight patients had endpoint events. The LAFI had the highest area under the receiver operating characteristic curve (AUC) predicting the endpoint events, with an AUC of 0.90 (95% CI 0.84-0.94). Multivariate Cox analysis demonstrated that only the LAFI (HR: 0.91, 95% CI 0.87-0.96, P < 0.0001) was independently predictive of endpoint events. Kaplan‒Meier survival curves showed that patients with an LAFI ≤ 42.25 cm/cc/m2 had more events than patients with an LAFI > 42.25 cm/cc/m2 (HR: 19.15, 95% CI 8.90-41.21, P < 0.001). The LAFI is a strong and independent predictor of events in STEMI patients treated with PCI.


Spatial and temporal localization of FGF-2 and VEGF in healing tooth extraction sockets in a rabbit model.
The purpose of this study was to test the hypothesis that spatial and temporal localization of growth factors FGF-2 and VEGF in a rabbit tooth extraction socket model correlate with the histologic events of healing. Twenty-four male New Zealand white rabbits divided into 8 groups of 3 were used in the study. Incisor teeth were extracted from both jaws and the healing extraction socket with surrounding jaw bone was harvested at 48 hours, 4 days, 1, 2, 4, 8, 12, and 16 weeks. Tissues were fixed, decalcified, and processed for hematoxylin-eosin and immunohistochemical staining. The sections were stained to detect FGF-2 and VEGF. The stained sections were then imaged and an automated computer program was used to detect the brown diaminobenzidine stain that represented the growth factors of interest. Data was obtained in the form of percentage area and intensity of stain and analyzed using the analysis of variance (ANOVA - Tukey Kramer and Scheffe's post-test). Spatial and temporal differences in localization of FGF-2 and VEGF were observed across all time frames in both jaws. Statistically significant differences in percentage area and intensity of brown diaminobenzidine stain were seen temporally between FGF-2 and VEGF (P < .05). The results of this study showed positive correlation of histologic events to spatial and temporal localization of FGF-2 and VEGF in a rabbit tooth extraction model.


Epigenome Maintenance in Response to DNA Damage.
Organism viability relies on the stable maintenance of specific chromatin landscapes, established during development, that shape cell functions and identities by driving distinct gene expression programs. Yet epigenome maintenance is challenged during transcription, replication, and repair of DNA damage, all of which elicit dynamic changes in chromatin organization. Here, we review recent advances that have shed light on the specialized mechanisms contributing to the restoration of epigenome structure and function after DNA damage in the mammalian cell nucleus. By drawing a parallel with epigenome maintenance during replication, we explore emerging concepts and highlight open issues in this rapidly growing field. In particular, we present our current knowledge of molecular players that support the coordinated maintenance of genome and epigenome integrity in response to DNA damage, and we highlight how nuclear organization impacts genome stability. Finally, we discuss possible functional implications of epigenome plasticity in response to genotoxic stress.


The recovery of mammalian cells treated with methyl methanesulfonate, nitrogen mustard or UV light. I. The effect of alkylation products on DNA replication.
CHO cells were synchronized in G1 phase and treated with MMS or HN2. The subsequent rate of DNA replication was found to be reduced in a dose-dependent manner. In addition, 2 X 10(-3 M and 3 X 10(-3) M MMS resulted in a 3--4 h delay prior to the initiation of S phase. If the cells were held for 8 h in hydroxyurea after MMS treatment, no subsequent lag in DNA synthesis was seen after removal of the hydroxyurea. The entry of confluent cells into S phase was found to be delayed 7 h upon trypsinizing and replating. Treatment of these cells with MMS resulted in a reduced rate of DNA replication, but no further delay in its initiation. Repair replication was found to continue at a constant rate for at least 12 h following MMS treatment of cells under all of these conditions. At the concentrations used in these experiments MMS severely inhibited the rate of protein synthesis, but HN2 had little effect. By comparing both the kinetics of repair replication and recovery of protein synthesis with the rate of DNA replication, it was concluded that the initial, severe reduction in rate following MMS treatment was probably due to an inhibition of protein synthesis.


Mode of action and antifungal properties of two cold-adapted chitinases.
The mode of action of two chitinases from the Antarctic Arthrobacter sp. strain TAD20 on N-acetyl-chitooligomers and chitin polymers has been elucidated. Identification of the length of chitin oligomers following enzymatic hydrolysis was verified by using HPLC-based analysis. It was observed that the length of the oligomer is important for enzyme action. The enzymes cannot effectively hydrolyze chitin oligomers with a degree of polymerization lower than four. ArChiA is an endochitinase which hydrolyzes chitin substrates randomly, whereas ArChiB is an exochitinase which degrades chitin chains and N-acetyl-chitooligomers from the nonreducing end, releasing N- N'-diacetyl-chitobiose. ArChiB (100 microg/ml) inhibited spore germination and hyphal elongation of the phytopathogenic fungus Botrytis cinerea by 15% and 30%, respectively. A more pronounced effect was observed with ArChiA (100 microg/ml) resulting in 70% inhibition of spore germination and 60% inhibition of germ tube elongation. A slight additive effect was observed, when the two enzymes were used in combination, only on the inhibition of germ tube elongation.


Australia's potentially most venomous snake: Parademansia microlepidotus.
Studies on the venom of the small-scaled or fierce snake, Parademansia microlepidotus, suggested that its combination of high toxicity and relatively large venom yield make it the most venomous snake in Australia. The species is distinct from the taipan, but its venom is neutralized by taipan antivenom but not other monospecific antivenoms.


Proteasome inhibition and aggresome formation in sporadic inclusion-body myositis and in amyloid-beta precursor protein-overexpressing cultured human muscle fibers.
The 26S proteasome system is involved in eliminating various proteins, including ubiquitinated misfolded/unfolded proteins, and its inhibition results in cellular accumulation of protein aggregates. Intramuscle-fiber ubiquitinated multiprotein-aggregates are characteristic of sporadic inclusion-body myositis (s-IBM) muscle fibers. Two major types of aggregates exist, containing either amyloid-beta (Abeta) or phosphorylated tau (p-tau). We have now asked whether abnormalities of the 26S proteasome contribute to s-IBM pathogenesis and whether the multiprotein aggregates have features of aggresomes. Using cultured human muscle fibers we also studied the effect of amyloid-beta precursor protein (AbetaPP) overexpression on proteasome function and the influence of proteasome inhibition on aggresome formation. We report that in s-IBM muscle biopsies 26S proteasome subunits were immunodetected in the gamma-tubulin-associated aggresomes, which also contained Abeta, p-tau, ubiquitin, and HSP70. In addition, a) expression of proteasome subunits was greatly increased, b) the 20Salpha proteasome subunit co-immunoprecipitated with AbetaPP/Abeta, and c) the three major proteasomal proteolytic activities were reduced. In cultured muscle fibers, AbetaPP-overexpressing fibers displayed diminished proteasomal proteolytic activities, and addition of proteasome inhibitor strikingly increased aggresome formation. Accordingly, proteasome dysfunction in s-IBM muscle fibers may play a role in accumulation of misfolded, potentially cytotoxic proteins and may be induced by increased intracellular AbetaPP/Abeta.


Genetic variants in autophagy-related genes and granuloma formation in a cohort of surgically treated Crohn's disease patients.
Granulomas are a characteristic microscopic finding in Crohn's disease. Their clinical significance is controversial and their pathogenesis is unknown, but impaired processing of bacterial components has been suggested. Autophagy is a fundamental process involved in the elimination of intracellular bacteria. Genetic variants in autophagy genes IRGM and ATG16L1 have been associated with susceptibility to Crohn's disease. We therefore investigated whether variants in autophagy genes contribute to granuloma formation. Surgical specimens from 464 clinically well-documented Crohn's patients were reviewed and scored for the presence and distribution of granulomas. All patients were genotyped for the CD-associated SNPs in ATG16L1 and IRGM as well as for 77 haplotype tagging SNPs in 13 additional autophagy genes. Granulomas were found in 75% of the patients. Their frequency increased with more distal involvement of the GI tract. Granuloma positive patients were significantly younger at the time of diagnosis and surgery, and were more likely to smoke. We identified associations between granulomas and autophagy gene variants ATG4A (rs5973822), FNBP1L (rs17109951) and ATG4D (rs7248026; rs2304165; rs10439163). These findings suggest that granuloma formation is a marker of a more aggressive disease course, and that variants in autophagy genes ATG4A, ATG2A, FNBP1L and ATG4D, may contribute to granuloma formation.


Targeting Cancer Metabolism and Cell Cycle by Plant-Derived Compounds.
Cancer has an increasing death rate over the world population without discriminating between the industrial and developing countries. Complexity of cancer such as ability of cancer cells to develop resistance to drugs or differential behavior of sub-types and different responses from the patients indicate the continuous need for research and development of new anticancer drugs, new formulations of drug combinations and treatment strategies. Not too surprisingly nature itself, is often the largest territorial reservoir as a source for this type of research and development. Speaking of plant variety, more than 1000 plants have already been identified to produce agents with anticancer activities. In this review, a panel of plant derived anti-cancer agents will be reiterated in terms of their mechanism of action in treatment of disease.


Comparison of different signal peptides for secretion of heterologous proteins in fission yeast.
In the fission yeast Schizosaccharomyces pombe, there are relatively few signal peptides available and most reports of their activity have not been comparative. Using sequence information from the S. pombe genome database we have identified three putative signal peptides, designated Cpy, Amy and Dpp, and compared their ability to support secretion of green fluorescent protein (GFP). In the comparison we also included the two well-described secretion signals derived from the precursors of, respectively, the Saccharomyces cerevisiae alpha-factor and the S. pombe P-factor. The capability of the tested signal peptides to direct secretion of GFP varied greatly. The alpha-factor signal did not confer secretion to GFP and all the produced GFP was trapped intracellular. In contrast, the Cpy signal peptide supported efficient secretion of GFP with yields approximating 10 mg/L. We also found that the use of an attenuated version of the S. cerevisiae URA3 marker substantially increases vector copy number and expression yield in fission yeast.


Locally Addressable Energy Efficient Actuation of Magnetic Soft Actuator Array Systems.
Advances in magnetoresponsive composites and (electro-)magnetic actuators have led to development of magnetic soft machines (MSMs) as building blocks for small-scale robotic devices. Near-field MSMs offer energy efficiency and compactness by bringing the field source and effectors in close proximity. Current challenges of near-field MSM are limited programmability of effector motion, dimensionality, ability to perform collaborative tasks, and structural flexibility. Herein, a new class of near-field MSMs is demonstrated that combines microscale thickness flexible planar coils with magnetoresponsive polymer effectors. Ultrathin manufacturing and magnetic programming of effectors is used to tailor their response to the nonhomogeneous near-field distribution on the coil surface. The MSMs are demonstrated to lift, tilt, pull, or grasp in close proximity to each other. These ultrathin (80 µm) and lightweight (100 gm-2 ) MSMs can operate at high frequency (25 Hz) and low energy consumption (0.5 W), required for the use of MSMs in portable electronics.


The azygos defibrillator lead for elevated defibrillation thresholds: implant technique, lead stability, and patient series.
Conventional insertion of implantable cardioverter-defibrillator (ICD) includes an evaluation of the defibrillation threshold (DFT). Implanting an ancillary defibrillation lead in the azygos vein has been introduced as a therapeutic option in patients with "high" DFT. This study reports the efficacy and stability of azygos defibrillation coils implanted for elevated DFTs. This is a retrospective review of seven consecutive patients with right and left pectoral, single- and dual-chamber, and biventricular ICDs and elevated DFTs, in whom an azygos defibrillation coil was introduced. Addition of an azygos defibrillator lead achieved a satisfactory safety margin during single energy defibrillation efficacy testing in four out of seven patients, with success at maximum device output in two patients. No satisfactory safety margin was achieved in the remaining patient, despite the further addition of a subcutaneous defibrillation coil. No change in lead position was observed over a mean radiographic follow-up of 8 months. No complications were noted during a mean follow-up of 14 months, including no deaths, and no ICD shocks. Implanting a defibrillation coil into the azygos vein is feasible and safe. In a majority of patients with failed defibrillation efficacy testing, adding an azygos coil achieves success on repeat testing. Therefore, this technique is one option for lowering the defibrillation threshold in patients who fail DFT testing of their ICD.


Nonadherence to anti-tuberculosis treatment, reasons and associated factors among pulmonary tuberculosis patients in the communities in Indonesia.
Tuberculosis (TB) is the world's major public health problem. We assessed the proportion, reasons, and associated factors for anti-TB treatment nonadherence in the communities in Indonesia. This national coverage cross-sectional survey was conducted from 2013 to 2014 with stratified multi-stage cluster sampling. Based on the region and rural-urban location. The 156 clusters were distributed in 136 districts/cities throughout 33 provinces, divided into three areas. An eligible population of age ≥15 was interviewed to find TB symptoms and screened with a thorax x-ray. Those whose filtered result detected positive followed an assessment of Sputum microscopy, LJ culture, and Xpert MTB/RIF. Census officers asked all participants about their history of TB and their treatment-defined Nonadherence as discontinuation of anti-tuberculosis treatment for <6 months. Data were analyzed using STATA 14.0 (College Station, TX, USA). Nonadherence to anti-TB treatment proportion was 27.24%. Multivariate analysis identified behavioral factors significantly associated with anti-TB treatment nonadherence, such as smoking (OR = 1.78, 95% CI (1.47-2.16)); place of first treatment received: government hospital (OR = 1.45, 95% CI:1.06-1.99); private hospital (OR = 1.93, 95% CI: 1.38-2.72); private practitioner (OR = 2.24, 95% CI: 1.56-3.23); socio-demographic and TB status included region: Sumatera (OR = 1.44, 95% CI: 1.05-1.98); other areas (OR = 1.84, 95% CI: 1.30-2.61); low level of education (OR = 1.60, 95% CI: 1.27-2.03); and current TB positive status (OR = 2.17, 95% CI: 1.26-3.73). Nonadherence to anti-TB drugs was highly related to the personal perception of the respondents, despite smoking, current TB status, a place for the first treatment, education, and region. The position of the first TB treatment at the private practitioner was significantly associated with the risk of Nonadherence to treatment.


Tumour reducing and anticarcinogenic activity of selected spices.
Tumour reducing activity of extracts of eight commonly used spices in India were studied in mice transplanted intraperitoneally with Ehrlich ascites tumour. Oral administration of extracts of black pepper, asafoetida, pippali and garlic could increase the percentage of life span in these mice by 64.7%, 52.9%, 47% and 41.1%, respectively. However intraperitoneal administration of spice extracts did not produce any significant reduction in tumour growth except for sesame (38.8%). Garlic extract and asafoetida extracts also inhibited two stage chemical carcinogenesis induced by 7,12 dimethyl benzanthracene and croton oil on mice skin with significant reduction in papiloma formation. These results indicate the potential use of spices as anti-cancer agents as well as anti-tumour promoters.


Relation between chemotherapy dose, oestrogen receptor expression, and body-mass index.
Clinicians often reduce chemotherapy doses when treating obese patients because of concerns about overdosing. We assessed dose-response according to body-mass index (BMI) and oestrogen receptor (ER) expression of the primary tumour in premenopausal patients with node-positive breast cancer treated with classical CMF (cyclophosphamide, methotrexate, and 5-fluorouracil). Obese patients were significantly more likely to receive a lower chemotherapy dose (<85% of expected dose) for the first course than were those with normal or intermediate BMI (39%vs 16%, p<0.0001). For obese patients and for the total population, reducing the dose of chemotherapy was associated with a significantly worse outcome for the ER-negative cohort (total population hazards ratio 85%vs <85% 0.68 [95% CI 0.54-0.86] for disease free survival; 0.72 [0.56-0.94] for overall survival) but not for the ER-positive cohort (1.16 [0.97-1.40] for disease-free survival; 1.16 [0.94-1.44] for overall survival) [interaction p values=0.0001 for disease-free survival and 0.0019 for overall survival]. Our findings suggest that for women with ER-absent or ER-low tumours, reduction in chemotherapy dose should be avoided.


Cloning and sequencing of the F gene of live attenuated Urabe Am9 mumps virus.
A cDNA clone encoding the entire F gene of the live attenuated mumps virus (MpsV) strain, Urabe Am9, has been isolated and sequenced. The F gene sequence shows significant homology with the one reported for the wild-type MpsV Miyahara strain.


Low prevalence of Helicobacter pylori infection in patients with gastric cancer.
The Helicobacter pylori (Hp) infection is a well-known carcinogenic factor. There are only limited data on the presence of Hp infection in gastric cancer in Romania. Cross-sectional study of the prevalence of Hp infection in gastric cancer. All cases of gastric cancer confirmed by gastroscopy in an endoscopy unit with national referrals were analysed. The Hp status was checked. The Hp + cases were studied compared with the Hp - cases. From 6768 gastroscopies 84 cases with gastric cancer were detected. Among these in 40 the Hp status was determined. 37 cases were carcinoma, 1 lymphoma, 2 cases without microscopical aspect of malignancy. In 8 cases (20%) Hp was found +. The majority of the patients with gastric cancer did not use PPIs. There were no demographic life style differences between the patients with gastric cancer who were Hp + and Hp -. Prevalence of the Hp infection in gastric cancer is lower than that reported in the literature. The investigation of Hp in gastric cancer should be done routinely. These data represent the starting point for a prospective study.


Surgical alternative to trabeculectomy.
Non-penetrating glaucoma surgery, represented by deep sclerectomy and viscocanalostomy, is an effective method to lower intraocular pressure (IOP) in glaucomatous patients. Both procedures reduce IOP by allowing aqueous humor drainage without opening the anterior chamber. Deep sclerectomy, similar to trabeculectomy, provides aqueous external filtration in the subconjunctival space. This technique, with the adjunctive use of implants, antimetabolites, and goniopuncture, may provide final IOP comparable to those obtained with trabeculectomy, but with less complications. Viscocanalostomy is less dependent on external filtration since it increases trabecular aqueous outflow facility by micro-disrupting Schlemm's canal walls and juxtacanalicular trabecular meshwork. This technique is very safe, but it provides higher final IOPs compared to trabeculectomy. Non-penetrating surgery should be therefore considered a surgical alternative to trabeculectomy in specific clinical cases.


Persistent eosinophilia among 21 historic cases of Strongyloides stercoralis infection: a case for routine follow-up and repeat treatment.
Strongyloidasis affects more than 100 million people worldwide and causes persisting infection. We retrospectively reviewed 21 cases of parasitologically confirmed Strongyloides stercoralis infection at our centre. We found four individuals who had eosinophilia (>0.5x109/litre) more than 6 mo after treatment. This may represent ongoing or relapsed infection. Our data support the need for continued follow-up of treated individuals, particularly those with abnormal host defences who are at risk of severe forms of the disease.


Apparent desensitization of NMDA responses in Xenopus oocytes involves calcium-dependent chloride current.
N-Methyl-D-aspartate (NMDA) receptors were expressed and studied in Xenopus oocytes injected with rat brain RNA. NMDA application elicits a rapid inward current that decays in several seconds to a relatively stable level. This decay is reportedly due to desensitization. However, we found the early transient component could be evoked more than once during a single application of NMDA, suggesting that the receptor did not actually desensitize. Removal of external Ca2+, replacement of Ca2+ with Ba2+, or intracellular injection of EGTA abolished the transient component. Furthermore, a variety of Cl- channel blockers nearly eliminated the transient component and inhibited the plateau current as well. We propose that a significant portion of the NMDA current recorded in oocytes is carried by a transient inward Cl- current triggered by Ca2+ influx through the NMDA receptor/channel.


Efficacy, safety and immunogenicity of a human rotavirus vaccine (RIX4414) in Hong Kong children up to three years of age: a randomized, controlled trial.
A phase III, double-blind, randomized, controlled trial was conducted in Hong Kong to evaluate the efficacy, safety and immunogenicity of a human rotavirus vaccine, RIX4414 (Rotarix) against severe rotavirus gastroenteritis in children up to three years of age. Healthy infants aged 6-12 weeks were enrolled between 08-December-2003 and 31-August-2005 and received two oral doses of either RIX4414 vaccine (N=1513) or placebo (N=1512) given 2 months apart. Vaccine efficacy was assessed from two weeks post-Dose 2 until the children were two and three years of age. Anti-rotavirus IgA seroconversion rate was calculated pre-vaccination and 1-2 months post-Dose 2 using ELISA (cut-off=20 U/mL) for 100 infants. Safety was assessed until the children were two years of age; serious adverse events (SAEs) were recorded throughout the study period. In children aged two and three years of life, vaccine efficacy against severe rotavirus gastroenteritis was 95.6% (95% CI: 73.1%-99.9%) and 96.1% (95% CI: 76.5%-99.9%), respectively. The seroconversion rate 1-2 months after the second dose of RIX4414 was 97.5% (95% CI: 86.8%-99.9%). At least one SAE was recorded in 439 and 477 infants who were administered RIX4414 and placebo, respectively (p-value=0.130). Six intussusception cases were reported (RIX4414=4; placebo=2) and none was assessed to be vaccine-related. RIX4414 was efficacious, immunogenic and safe in the prevention of rotavirus gastroenteritis for at least two years post-vaccination in Hong Kong children.


Effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin on trace elements, inflammation and viral clearance in the myocardium during coxsackievirus B3 infection in mice.
A myocarditic coxsackievirus B3 (CB3) infection in adult male A/J mice was used to investigate the effects of 2,3,7,8-tetrachlorodibenzo-para-dioxin (TCDD) exposure on mortality and on inflammatory lesion, virus and trace element contents of the heart. The mice were injected with four weekly intraperitoneal (i.p.) injections of TCDD (a loading dose of 5 microg/kg followed by three maintenance doses of 1.4 microg/kg). To reach a steady-state body burden of TCDD the mice were allowed a 90-day recovery period before infection with CB3 virus. TCDD increased the infection-induced mortality rate, whereas in TCDD-exposed mice, heart lesions at day 7 after the virus inoculation (median value 0.67% of the tissue section area; interquartile range 0.28; not statistically significant) were one-third of that in non-exposed infected mice (2.07% of the tissue section area; interquartile range 3.06). The size of the inflammatory heart lesion correlated to the amount of virus (r(s) = 0.829, P < 0.01) as well as to the calcium (Ca: r(s) = 0.725, P < 0.01) and the magnesium (Mg: r(s) = -0.615, P < 0.05) contents. In TCDD-exposed mice in situ hybridisation of viral RNA in the myocardium at day 7 showed a tendency to decreased amounts of virus, as well as a less pronounced increase in myocardial Ca content, both supporting a milder myocardial disease after TCDD exposure. No effect of TCDD exposure was seen on the zinc (Zn) or selenium (Se) levels in the myocardium. In conclusion, although TCDD seemed to have a limiting effect on viral replication and the development of the inflammatory lesion in the myocardium, mortality was increased by TCDD in this infection model. However, TCDD had no significant effects on the selected trace elements that could be of importance for the severity of the inflammatory lesion (Ca, Se), for the local host response activation (Zn) or for the development of myocardial disease complications (Mg). Accordingly, the increased mortality may be a result of an infection-induced increase in TCDD toxicity to vital organs other than the heart, and/or a TCDD-induced change in the tissue affinity and virulence of the virus, possibly causing involvement of other target organs in the infectious process and changed pathogenesis.


Riboflavin and ultraviolet light: impact on dengue virus infectivity.
Dengue viruses (DENV 1-4) are emerging across the world, and these viruses pose a risk to transfusion safety. Pathogen inactivation may be an alternative approach for managing the risk of DENV transfusion transmission. This study aimed to investigate the ability of riboflavin and UV light to inactivate DENV 1-4 in platelet concentrates. DENV 1-4 were spiked into buffy coat-derived platelet concentrates in additive solution (SSP+) before being treated with riboflavin and UV light. Infectious virus was quantified pre- and posttreatment, and the reduction in viral infectivity was calculated. All four DENV serotypes were modestly reduced after treatment. The greatest amount of reduction in infectivity was observed for DENV-4 (1·81 log reduction) followed by DENV-3 (1·71 log reduction), DENV-2 (1·45 log reduction) and then DENV-1 (1·28 log reduction). Our study demonstrates that DENV 1-4 titres are modestly reduced following treatment with riboflavin and UV light. With the increasing number of transfusion-transmitted cases of DENV around the globe, and the increasing incidence and geographical distribution of DENV, additional approaches for maintaining blood safety may be required in the future.


Electroencephalography for predicting antidepressant treatment success: A systematic review and meta-analysis.
Patients suffering from major depressive disorder (MDD) regularly experience non-response to treatment for their depressive episode. Personalized clinical decision making could shorten depressive episodes and reduce patient suffering. Although no clinical tools are currently available, machine learning analysis of electroencephalography (EEG) shows promise in treatment response prediction. With a systematic review and meta-analysis, we evaluated the accuracy of EEG for individual patient response prediction. Importantly, we included only prediction studies that used cross-validation. We used a bivariate model to calculate prediction success, as expressed by area-under the curve, sensitivity and specificity. Furthermore, we analyzed prediction success for separate antidepressant interventions. 15 studies with 12 individual patient samples and a total of 479 patients were included. Research methods varied considerably between studies. Meta-analysis of results from this heterogeneous set of studies resulted in an area under the curve of 0.91, a sensitivity of 83 % (95 % CI 74-89 %), and a specificity of 86 % (95 % CI 81-90 %). Classification performance did not significantly differ between treatments. Although studies were all internally validated, no externally validated studies have been reported. We found substantial risk of bias caused by methodological shortcomings such as non-independent feature selection, though performance of non-biased studies was comparable. Sample sizes were relatively small and no study used external validation, increasing the risk of overestimation of accuracy. Electroencephalography can predict the response to antidepressant treatment with high accuracy. However, future studies with more rigorous validation are needed to produce a clinical tool to guide interventions in MDD. CRD42021268169.


Children using a unilateral cochlear implant and contralateral hearing aid: bimodal hearing outcomes when one ear is outside the UK (NICE 2009) audiological criteria for cochlear implantation - a single site case-control study.
In the new revised National Institute for Health & Care Excellence (NICE, TA566, 2019) guidelines for cochlear implantation (CI) have clearly stipulated that the hearing loss must be bilateral. Prior to this revision, children and young people (CYP) with asymmetrical thresholds have been considered for unilateral CI when one ear was in audiological criteria. Children with asymmetrical hearing loss represent an important cohort of potential CI candidates, who will continue to be prevented from benefiting from CI unless evidence is produced to support implantation and maximise subsequent benefit.The aim of this study is to evaluate the 'real-life' hearing performance in a group of children who have received a unilateral CI and who have hearing thresholds in the contralateral ear that are outside the current UK NICE 2019 audiological criteria for CI. The contralateral ear will be aided using a conventional hearing aid (HA). The outcomes from this 'bimodal' group will be compared with a group of children who have received bilateral CI, and a group of children using bilateral HA, to extend the current knowledge about the different performance levels between bilateral CI, bilateral HA and bimodal hearing in CYP. Thirty CYP aged 6-17 years old, 10 bimodal users, 10 bilateral HA users and 10 bilateral cochlear implant users will be subjected to a test battery consisting of: (1) spatial release from masking, (2) complex pitch direction discrimination, (3) melodic identification, (4) perception of prosodic features in speech and (5) TEN test. Subjects will be tested in their optimal device modality. Standard demographic and hearing health information will be collected. In the absence of comparable published data to power the study, sample size was determined on pragmatic grounds. Tests are exploratory and for hypothesis generating purposes. Therefore, the standard criterion of p<0.05 will be used. This has been approved by the Health Research Authority and NHS REC within the UK (22/EM/0104). Industry funding was secured via a competitive researcher-led grant application process. Trial results will be subject to publication according to the definition of the outcome presented in this protocol.


Association of MC4R (rs17782313) with diabetes and cardiovascular disease in Korean men and women.
Diabetes is mostly assessed by the fasting glucose level. Several studies reported that serum fasting glucose levels and cardiovascular disease are associated with MC4R. A total of 4294 subjects participated in this study. There were 1810 subjects with cardiovascular disease among the 4294 subjects. We used multivariate linear regression models and multiple logistic regression analysis. Individuals with the TC/CC genotype had a 1.29-fold higher risk of diabetes than did those with the TT genotype when adjusting for age, sex, and BMI (OR, 1.29; 95% CI, 1.04-1.60). For healthy subjects, the association was significant in women (OR, 1.99; 95% CI, 1.01-3.93). Men with the TC/CC genotype had a 1.21-fold higher risk of cardiovascular disease than did those with the TT genotype when adjusting for age, sex, and BMI (OR, 1.21; 95% CI, 1.04-1.41). The relationship between MC4R and cardiovascular disease was stronger in lean men (OR, 1.40; 95% CI, 1.12-1.74, p = 0.0028) than in overweight men. This study suggests that the rs17782313 SNP in MC4R is related to diabetes and the SNP is also associated with cardiovascular disease in lean men.


Decidualization modulates a signal transduction system via protease-activated receptor-1 in endometrial stromal cells.
Decidual cells are thought to be involved in the maintenance of pregnancy. We conducted this study to evaluate the cellular function of endometrial stromal cells (ESCs) transitioning to decidualization. Normal endometrial specimens were obtained from premenopausal patients who had undergone hysterectomies for subserosal leiomyomas. Decidualization of the ESCs (DSCs) was induced by incubating subconfluent cells in media containing medroxyprogesterone acetate and dibutyryl-cyclic adenosine monophosphate. We first analyzed the expression profile of protease-activated receptor-1 (PAR-1) between ESCs and DSCs. To investigate the intracellular signal transduction system in the DSCs, we incubated cells with thrombin receptor activator peptide 6 (TRAP-6). The levels of IL-8, monocyte chemo-attractant protein-1, matrix metalloproteinase (MMP)-1, and vascular endothelial growth factor in the culture medium were measured by enzyme-linked immunosorbent assays. The activation of the MAP kinase signaling pathway was detected by a Western blot analysis. The activation was evaluated for the expression of p21. PAR-1 receptor expression is upregulated in DSCs. The productions of chemokine and MMP-1 increased in the DSCs with the addition of TRAP-6. The activity of both the ERK-1 and ERK-2 isoforms was increased by 5-15 minute after TRAP-6 treatment. p70 S6 kinase showed the strongest expression after 1 hour. p21 was strongly observed in ESCs compared to the DSCs. Our results suggest that cell function is changed by decidualization in association with increasing PAR-1 expression. The upregulation of PAR-1 may have some influence on pregnancy in the decidua.


Nanomechanical clues from morphologically normal cervical squamous cells could improve cervical cancer screening.
Applying an atomic force microscope, we performed a nanomechanical analysis of morphologically normal cervical squamous cells (MNSCs) which are commonly used in cervical screening. Results showed that nanomechanical parameters of MNSCs correlate well with cervical malignancy, and may have potential in cancer screening to provide early diagnosis.


Pulmonary clearance and toxicity of zinc oxide instilled into the rat lung.
Pulmonary toxicity of zinc oxide (ZnO) was evaluated by investigating the metabolic fate and inflammatory potency of ZnO instilled into the rat lung. Groups of three rats received single intratracheal instillations of ZnO suspension at a dose of 100 micrograms Zn/rat in the time-course experiment or received 20, 50, 100, 200, 500 and 1000 micrograms Zn/rat and were killed 2 days after treatment in the dose-response experiment. It was suggested that ZnO particles were solubilized in the bronchoalveolar milieu and cleared from the lung with a half-life of 14 h. Metallothionein (MT) was induced with a peak at 2 days. The content of MT was proportional to the dose of ZnO, but contributed little for the accumulation of Zn in the lung. A dose of 20 micrograms Zn/rat was sufficient to develop maximum responses for beta-glucuronidase activity and surfactant content in the bronchoalveolar lavage fluid. Moreover, the activity of lactate dehydrogenase and protein content in the lavage fluid increased significantly at 20 micrograms Zn/rat. These results suggest that the recommended ZnO concentration in the work-place atmosphere of 5 mg/m3 might not be adequate.


Phenol degradation by Fenton-like process.
The main objective of this study was to investigate the optimum conditions for the Fenton-like process on phenol degradation, using Mn(2+) as a supporting catalyst in the Fenton reaction. The effect of the independent factors [H2O2], [Fe(2+)], [Mn(2+)] and t (reaction time) was evaluated on the efficiency of phenol degradation at two pHs (3 and 5). The experimental arrangement adopted was the Box-Behnken delineation, with the phenol concentration after the treatments suggested as response variable. At less acidic pH (5), regardless of [Mn(2+)], it was observed that the conventional Fenton process was the most efficient alternative, considering the optimum condition: 2.65 mmol L(-1) for [H2O2], 0.36 mmol L(-1) for [Fe(2+)], and 90 min for t. It was observed that the addition of Mn(2+) helped the phenol degradation at more acidic pH (3), obtaining the optimum condition: 6.17 mmol L(-1) for [H2O2], 0.36 mmol L(-1) for [Fe(2+)], 1.09 mmol L(-1) for [Mn(2+)], and 90 min for t.


Atrial fibrillation/flutter in myopathies.
With improved screening of patients with primary and secondary myopathies and more comprehensive investigations it turns out that an increasing number of patients with myopathies develops cardiac disease (cardiac involvement), before or after onset of the neuromuscular abnormalities. Cardiac involvement in myopathies manifests within the myocardium or the cardiac conduction system with impulse generation or conduction disturbances. An increasingly recognized rhythm abnormality in these patients is atrial fibrillation/flutter (AFI/AFL), which carries an increased risk for stroke embolism and represents an absolute indication for oral anticoagulation (OAC). Primary myopathies, in which AFI/AFL has been described so far include dystrophinopathies, Emery-Dreifuss muscular dystrophy, facio-scapulo-humeral muscular dystrophy, limb girdle muscular dystrophies, congenital myopathies, myofibrillar myopathies, myotonic dystrophies, glycogenoses, mitochondrial disorders, Barth syndrome, McLeod syndrome, and non-specific myopathies. Secondary myopathies, in which AFI/AFL has been described comprise polymyositis, dermatomyositis, colchicine-induced myopathy, and hyperthyroid myopathy. Myopathies most commonly associated with AFI/AFL are myotonic dystrophy and Emery-Dreifuss muscular dystrophy. Information about the frequency of stroke/embolism in these patients is rudimentary but there are indications that it is not increased in these patients. Only a few patients with AFI/AFL receive OAC to prevent from stroke/embolism. Patients with myopathy and AFI/AFL require thorough surveillance. If additional cardiovascular risk factors develop, OAC should be considered as in patients with other causes of AFI/AFL.


Multiquanta flux jumps in superconducting fractal.
We study the magnetic field response of millimeter scale fractal Sierpinski gaskets (SG) assembled of superconducting equilateral triangular patches. Directly imaged quantitative induction maps reveal hierarchical periodic filling of enclosed void areas with multiquanta magnetic flux, which jumps inside the voids in repeating bundles of individual flux quanta Φ0. The number Ns of entering flux quanta in different triangular voids of the SG is proportional to the linear size s of the void, while the field periodicity of flux jumps varies as 1/s. We explain this behavior by modeling the triangular voids in the SG with effective superconducting rings and by calculating their response following the London analysis of persistent currents, Js, induced by the applied field Ha and by the entering flux. With changing Ha, Js reaches a critical value in the vertex joints that connect the triangular superconducting patches and allows the giant flux jumps into the SG voids through phase slips or multiple Abrikosov vortex transfer across the vertices. The unique flux behavior in superconducting SG patterns, may be used to design tunable low-loss resonators with multi-line high-frequency spectrum for microwave technologies.


The association between body lead levels and childhood rickets: A meta-analysis based on Chinese cohort.
China has serious lead pollution and a high incidence of childhood rickets. High lead levels have been reported in childhood rickets, but the results were inconsistent.To evaluate the association between body lead levels and childhood rickets.After a systematic literature search, we identified 15 studies determining body lead levels between rickets children and healthy controls, and 4 studies focusing on the cases of different disease severity. Standard mean differences (SMD) and the corresponding 95% confidence intervals (CI) were pooled to compare the lead levels between different groups.Sixteen case-control studies were included with a total of 5082 cases and 6054 controls. Compared with healthy controls, the body lead levels in rickets children were significantly higher (SMD (95%CI): 0.67 (0.41-0.93)), and subgroup analyses showed consistent results. The cases with moderate-to-severe disease activity also had a significantly higher lead level than mild-to-moderate cases (SMD (95%CI): 0.64 (0.31-0.97)).This meta-analysis suggested an association between body lead levels and childhood rickets, and lead exposure might be a risk factor for rickets.


Nonlinear analysis of dynamic binding in affinity capillary electrophoresis demonstrated for inclusion complexes of beta-cyclodextrin.
The stability of the molecular host-guest inclusion complexes of beta-cyclodextrin with benzoate and four different hydroxybenzoates is investigated. For the measurement of the binding constants an experimental method is devised that is based on affinity capillary electrophoresis (ACE) with indirect UV absorbance detection. We derive an explicit equation for effective mobilities in ACE experiments without violation of rigorous mass balance. This equation is employed in the nonlinear least-squares analyses of the experimental data yielding binding constants of 48+/-2 M(-1) for benzoate, 299+/-38 M(-1) for 2-hydroxybenzoate, 37+/-1 M(-1) for 3-hydroxybenzoate, 228+/-9 M(-1) for 4-hydroxybenzoate, and 895+/-110 M(-1) in the case of 2,4-dihydroxybenzoate.


Elongation inhibition by DRB sensitivity-inducing factor is regulated by the A20 promoter via a novel negative element and NF-kappaB.
A20 is an immediate-early NF-kappaB target gene. Prior to NF-kappaB stimulation, the A20 promoter is bound by the polymerase II machinery to allow rapid transcription activation. Here we show that the basal A20 transcription is repressed at the level of elongation in a promoter-specific fashion. Immunodepletion in vitro and RNA interference in cultured cells suggest that the basal elongation inhibition is conferred by DRB sensitivity-inducing factor (DSIF). We have identified a negative upstream promoter element called ELIE that controls DSIF activity. Remarkably, following NF-kappaB stimulation, inhibition of the A20 promoter by DSIF persists, but it is now regulated by NF-kappaB rather than ELIE. Similar regulation by DSIF is shown for another NF-kappaB-responsive gene, the IkappaBalpha gene. These findings reveal an intimate and dynamic relationship between DSIF inhibition of elongation and promoter-bound transcription factors. The potential significance of the differential regulation of DSIF activity by cis-acting elements is discussed.


DNA visualization in single molecule studies carried out with optical tweezers: Covalent versus non-covalent attachment of fluorophores.
In this study, we investigated the use of the covalent attachment of fluorescent dyes to double-stranded DNA (dsDNA) stretched between particles using optical tweezers (OT) and compared the mechanical properties of the covalently-functionalized chain to that of unmodified DNA and to DNA bound to a previously uncharacterized groove-binder, SYBR-gold. Modified DNA species were obtained by covalently linking azide-functionalized organic fluorophores onto the backbone of DNA chains via the alkyne moieties of modified bases that were incorporated during PCR. These DNA molecules were then constructed into dumbbells by attaching polystyrene particles to the respective chain ends via biotin or digoxigenin handles that had been pre-attached to the PCR primers which formed the ends of the synthesized molecule. Using the optical tweezers, the DNA was stretched by separating the two optically trapped polystyrene particles. Displacements of the particles were measured in 3D using an interpolation-based normalized cross-correlation method and force-extension curves were calculated and fitted to the worm-like chain model to parameterize the mechanical properties of the DNA. Results showed that both the contour and persistence length of the covalently-modified dsDNAs were indistinguishable from that of the unmodified dsDNA, whereas SYBR-gold binding perturbed the contour length of the chain in a force-dependent manner.


Mirror therapy in lower limb amputees--a look beyond primary motor cortex reorganization.
Phantom pain in upper limb amputees is associated with the extent of reorganization in the primary sensorimotor cortex. Mirror visual feedback therapy has been shown to improve phantom pain. We investigated the extent of cortical reorganization in lower limb amputees and changes in neural activity induced by mirror therapy. Eight lower limb amputees underwent 12 sessions of MVFT and functional magnetic resonance imaging (fMRI) of the brain before the first and after the last MVFT session. FMRI sessions consisted of two runs in which subjects were instructed to perform repetitive movement of the healthy and phantom ankle. Before MVFT, the mean phantom pain intensity was 4.6 ± 3.1 on a visual analog scale and decreased to 1.8 ± 1.7 (p = 0.04). We did not observe a consistent pattern of cortical activation in primary sensorimotor areas during phantom limb movements. Following MVFT, increased activity was obtained in the right orbitofrontal cortex during phantom ankle movements. Comparison of cortical activity during movements of the phantom ankle and the intact ankle showed significantly higher activity in the left inferior frontal cortex (pars triangularis). These results question the known association between phantom pain and primary sensorimotor reorganization and propose reorganizational changes involving multiple cortical areas in lower limb amputees. Finally, reduction of phantom pain after mirror visual feedback therapy was associated with increased prefrontal cortical activity during phantom ankle movements.


Sexual Activity After Bariatric Surgery: A Prospective Monocentric Study Using the PISQ-IR Questionnaire.
Obesity can lead to pelvic floor disorders, interfering with women's sexual life; Prolapse/Incontinence Sexual Questionnaire-International Urogynecology Association-Revised (PISQ-IR) is a new instrument to measure sexual life in women with pelvic floor disorders. To assess the utility of using PISQ-IR in morbidly obese women undergoing bariatric surgery and to show the improvement of bariatric surgery on sexuality. This prospective monocentric study included all women who underwent bariatric surgery from June 2016-May 2017. Sleeve gastrectomy (SG) or Roux-en-Y gastric bypass (RYGB) was performed by 1 surgeon, and data were collected by 1 researcher. Demographics, medical history, number of deliveries, and type of bariatric surgery (SG or RYGB) were collected at baseline. At the 1-year follow-up consultation, postoperative complications, percentage of total body weight loss (TWL) and percentage excess weight loss were assessed. Questionnaires were given at baseline and at 1-year follow-up. Postoperative complications and total weight loss were measured at the 1-year follow-up. Sexual activity, using the PISQ-IR, and pelvic organ prolapse, urinary incontinence, and anal incontinence, using the urinary symptom profile and PFDI (Pelvic Floor Distress Inventory) 20, were compared before surgery and at 1-year follow-up. 72 patients were included at baseline, 54 (75%) were considered for final analysis (30 RYGB and 24 SG), and 51 patients were considered for PISQ-IR. The mean preoperative body mass index was 41 ± 5.4 kg/m2, and mean age was 43 ± 11.8 years. Both procedures induced an important weight loss (mean TWL of 33%) and had a similar percentage of TWL for each procedure: 31 (15-46) for SG vs 34 (9-51) for RYGB. 9 patients (17 [6%]) became sexually active 1 year after surgery. For the sexually inactive population after 1 year, there was no differences in the PISQ-IR in the follow-up. When body mass index loss was >13 kg/m2, a higher percentage of the sexually active population improved their different scores, and there were significant results for both global quality of life and desire (P = .026 and .046). The other questionnaires showed a significant decrease in urinary incontinence symptoms (P < .001) associated with weight loss, whereas there was no difference in terms of pelvic organ prolapse or anal incontinence. PISQ-IR is a useful instrument to measure sexual function regarding pelvic floor disorders. Bariatric surgery improves sexual activity in obese women 1 year after surgery. Treacy PJ, Mazoyer C, Falagario U, et al. Sexual Activity After Bariatric Surgery: A Prospective Monocentric Study Using the PISQ-IR Questionnaire. J Sex Med 2019;16:1930-1937.


Interaction of tubulin with phospholipid vesicles. I. Association with vesicles at the phase transition.
Purified soluble tubulin will interact with highly sonicated small unilamellar vesicles of dipalmitoyl phosphatidylcholine. This results in the leakage of a fluorescent dye, carboxyfluorescein, from the internal aqueous space of the vesicles at the lipid phase transition. The amount of dye leaked from the vesicles is determined by the tubulin to vesicle ratio and partial dye leakage reflects the uniform leakage of carboxyfluorescein from all of the vesicles. Once tubulin interacts with dipalmitoyl phosphatidylcholine vesicles at the phase transition, it is unavailable to interact with additional vesicles. This interaction represents the formation of stable tubulin-vesicle recombinants as shown by density gradient centrifugation. Such recombinants are only formed by interacting the protein with the vesicles at the lipid phase transition temperature. The interaction is insensitive to ionic strength (0.001 to 4 M NaCl) and the recombinants remain stable for days.


Enzymatic properties of starfish phospholipase A2 and its application.
Industrial phospholipase A2 (PLA2) mainly produced from porcine pancreas is used for production of lysolecithin which is well known as an excellent natural emulsifier for food, cosmetic, and pharmaceutical industries. Since the outbreak of bovine spongiform encephalopathy (BSE) or religious tradition, it is hoped that the new sources of PLA2, as well as other enzymes and proteins, will be developed instead of mammal. From these backgrounds, we studied for PLA2 from marine organisms and found that starfish Asterina pectinifera PLA2 possesses extremely high activity and characteristic polar-group specificity comparing with commercially available PLA2 from porcine pancreas. Therefore, it was suggested that the starfish A. pectinifera would be a potential source of PLA2, and the PLA2 can be utilized as alternative enzyme of mammalian PLA2.


Yams (Dioscorea spp.) from the South Pacific Islands contain many novel badnaviruses: implications for international movement of yam germplasm.
Yam (Dioscorea spp.) samples (n = 690) from seven South Pacific Islands were screened for badnavirus infection by ELISA using two antisera to African badnaviruses. Positive readings were obtained for 26.4-34.6% of samples representing both known (D. bulbifera, D. nummularia and D. pentaphylla) and unreported host species (D. alata, D. esculenta, D. rotundata and D. trifida) in this region. Total DNAs were extracted from 25 ELISA-positive plants and 4 ELISA-negative controls and subjected to PCR amplification with badnavirus-specific primers targeting the reverse transcriptase (RT)-RNaseH genes. All 29 samples yielded the expected size PCR-product for badnaviruses, which were cloned and sequenced. Phylogenetic analyses of the resulting 45 partial (500-527 bp) RT-RNaseH sequences revealed 11 new sequence groups with <79% nucleotide identity to each other or any EMBL sequence. Three sequences (two groups) were highly divergent to the other nine new South Pacific yam badnavirus groups (47.9-57.2% identity) and probably represent either new Caulimoviridae genera or endogenous pararetrovirus sequences. Some sequence groups appeared specific to particular Dioscorea host species. Four 99.9% identical RT-RNaseH sequences possessing nine amino acid deletions from D. esculenta from three islands represent a putative integrated sequence group. The distribution of sequence groups across the islands indicates that badnaviruses have spread extensively between islands and continents through infected germplasm.


Inhibitory effect of fluoxetine on lymphoma growth through the modulation of antitumor T-cell response by serotonin-dependent and independent mechanisms.
Fluoxetine, a selective serotonin reuptake inhibitor, is widely used for the treatment of depressive symptoms of cancer patients. However, there are contradictory evidences about its effects on immunity and cancer. Thus, we studied the effects of fluoxetine on tumor growth and on antitumoral T-cell-mediated immunity. In vivo chronic fluoxetine treatment inhibited tumor growth, and increased latency of appearance of solid tumors and survival of mice. Fluoxetine administration also increased mitogen-induced T-cell proliferation and Tumor Necrosis Factor-alpha (TNF-alpha) and Interferon-gamma (IFN-gamma) expression, without altering CD4(+)/CD8(+) ratio. In vitro, fluoxetine did not affect tumor cells proliferation, but it exerted a direct effect on T lymphocytes. Both fluoxetine and serotonin stimulated proliferation induced by a suboptimal mitogen concentration but inhibited proliferation at the optimal one. When both drugs were combined the results indicated that the effects of fluoxetine are in part independent of its ability to elevate serotonin extracellular levels. Finally, continue fluoxetine administration in nude mice - devoid of T lymphocytes - did not modify tumor progression, thus supporting the hypothesis of an immuno-modulatory effect of this drug on T cells that drives tumor growth control. These findings indicate, for the first time, that fluoxetine inhibits tumor growth through modulation of T-cell-mediated immunity by the already known serotonin-dependent pathway and by a novel independent mechanism.


Asians and Asian Americans' social media use for coping with discrimination: A mixed-methods study of well-being implications.
Although social media has the potential to serve as a coping tool, it is unclear how Asians and Asian Americans use social media to cope with discrimination, and how social media coping is associated with these users' psycho-emotional well-being. Drawing on the Multidimensional Model of Social Media Use, this mixed-methods study examined the well-being implications of three social media coping activities among Asians and Asian Americans. A total of 931 Asians and Asian Americans in the U.S. between the ages of 18 and 93 (M = 46.49, SD = 16.58; 49.2% female) completed an anonymous online survey. Twenty-three participants (12 females) between the ages of 19 and 70 joined seven focus group interviews. Survey results showed that messaging was related to greater race-based traumatic stress (RBTS), although it also had a weak indirect relationship with positive emotions through social support. Posting and commenting had an indirect relationship with both lower RBTS and greater positive emotions via social support. Reading and browsing was associated with greater RBTS (directly) and positive emotions (both directly and indirectly through social support). Focus group data revealed how the three activities contributed to perceived social support and why these activities were associated with promising and/or concerning well-being outcomes.


Clinical and pathological characteristics of mycobacterial tenosynovitis and arthritis.
To investigate the clinical characteristics and pathological features of patients with mycobacterial tenosynovitis and arthritis. All patients with tenosynovitis and arthritis caused by Mycobacterium tuberculosis (MTB) and nontuberculous mycobacteria (NTM) who were treated at a medical center in Taiwan from 2001 to 2010 were analyzed. Thirty-two patients with mycobacterial tenosynovitis and arthritis were identified. MTB was isolated exclusively from patients with arthritis of large joints (n = 11), while NTM were isolated from patients with arthritis of large joints (n = 4) and from those with tenosynovitis (n = 17). Among patients with tenosynovitis due to NTM, the most commonly found NTM were M. marinum (n = 7), M. intracellulare (n = 5), and M. abscessus (sensu stricto) (n = 2). Six of the seven patients with tenosynovitis due to M. marinum had suffered fishing-related injuries to the hands. All four patients with NTM arthritis had recurrent septic arthritis after surgery. NTM were isolated once from the debrided tissue specimens in three of these patients; the other patient died of systemic infection caused by M. intracellulare and multiple bacterial pathogens. Mycobacterial tenosynovitis should be considered in patients who present with indolent symptoms of chronic tenosynovitis. Complete clinical information, including history of trauma or joint replacement surgery and underlying systemic disease, is helpful in establishing an early diagnosis of the disease.


[Relationship between the prognosis of retinal detachment in emmetropic eyes and vitreous changes].
To study the relationship between surgical prognosis of retinal detachment (RD) in emmetropic or myopic eyes and vitreous changes. Postoperatively, 1,294 eyes of 1,262 patients with rhegmatogenous RD including 243 emmetropic and 1,031 myopic eyes were retrospectively studied. The affects of proliferative vitreoretinopathy (PVR), posterior vitreous detachment (PVD), vitreous liquefaction and concentration on the surgical prognosis were evaluated. The cure rate in emmetropic group (88.5%) was lower than that in myopic group (93.8%, P < 0.01). Also, the cure rate by initial surgery in emmetropic group (65.4%) was lower than that in myopic group (76.4%, P < 0.01). In addition, the occurrences of PVD (39.6%) and liquefaction (76.1%) in emmetropic group were lower than that in myopic group (51.7%, and 81.8% respectively, P < 0.05). However, focal vitreous concentration in emmetropic group (56.8%) was much higher than that in myopic group (19.6%, P < 0.01). The surgical prognosis of RD especially by initial operation is worse in emmetropic group than that in myopic group. The lower occurrences of PVD and liquefaction but higher occurrence of focal vitreous concentration in emmetropic eyes may have impact on the prognosis. It is possible that the vitreous characteristics in emmetropic eyes with RD have relationship to the occurrence and development of PVR that has the influence on the prognosis to some extent.


Spatial patterns of mitochondrial and nuclear gene pools in chamois (Rupicapra r. rupicapra) from the Eastern Alps.
We have assessed the variability of maternally (mtDNA) and biparentally (allozymes) inherited genes of 443 chamois (Rupicapra r. rupicapra) from 19 regional samples in the Eastern Alps, to estimate the degree and patterns of spatial gene pool differentiation, and their possible causes. Based on a total mtDNA-RFLP approach with 16 hexanucleotide-recognizing restriction endonucleases, we found marked substructuring of the maternal gene pool into four phylogeographic groups. A hierarchical AMOVA revealed that 67.09% of the variance was partitioned among these four mtDNA-phylogroups, whereas only 8.04% were because of partitioning among regional samples within the populations, and 24.86% due to partitioning among individuals within regional samples. We interpreted this spatial pattern of mtDNA variability as a result of immigration of chamois from different Pleistocene refugia surrounding the Alps after the withdrawal of glaciers, rather than from topographic barriers to gene flow, such as Alpine valleys, extended glaciers or woodlands. However, this striking geographical structuring of the maternal genome was not paralleled by allelic variation at 33 allozyme loci, which were used as nuclear DNA markers. Wright's hierarchical F-statistics revealed that only < or =0.45% of the explained allozymic diversity was because of partitioning among the four mtDNA-phylogroups. We conclude that this discordance of spatial patterns of nuclear and mtDNA gene pools results from a phylogeographic background and sex-specific dispersal, with higher levels of philopatry in females.


Discovery in toxicology: mediation by gene expression array technology.
Toxicogenomics is a term that represents the merging of toxicology with novel genomics techniques. Data generated in the new-age era of toxicology is relatively complex, requires new bioinformatics tools for adequate interpretation, and allows for the rapid generation of testable hypotheses. Hazard identification and risk assessment processes will advance from the use of genomics techniques, which will lead to greater understanding of mechanism(s) of action of toxicants, development of novel biomarkers of exposure and effect, and better identification of sensitive subpopulations.


Signal processing properties of fast spiking interneurons.
Fast spiking interneurons receive excitatory synaptic inputs from pyramidal cells and a relevant problem is to understand how these cells readout this information. Here this topic is investigated theoretically by using a biophysical modeling of a pair of coupled fast spiking interneuron models. The model predicts, in agreement with the experimental findings, that these cells are capable of transmitting pre-synaptic signals with high temporal precision and transferring high frequency inputs while preserving their relative timing. Moreover, it is shown that a pair of fast spiking interneurons, coupled through both inhibitory and electrical synapses, behaves as a coincidence detector. Lastly, to understand the mechanisms underlying these phenomena, a theoretical analysis is carried out by using a simpler modeling approach.


EEG diagnoses of neonatal seizures: clinical correlations and outcome.
Electroencephalographic seizures were evaluated in 112 neonates. The first portion of the study involved 80 neonates with clinically identified abnormal movements, 8 of whom (10%) had electroencephalographic evidence of seizures coincident with this activity. Patients with abnormal movements (90%) had no concurrent electrical seizures. In the second part of the study, 40 infants who had electrical seizures were investigated. Eight of these infants had been identified during the first part of the study. Two-thirds of the patients (25) were premature. Sixteen patients (40%) died; 90% had brain lesions documented by computed tomography and/or postmortem study. Cerebral infarction and intraventricular hemorrhage were the most common lesions. One-third of the survivors (8 of 24 patients) were normal at a mean age of 3 years, while two-thirds had significant neurologic and developmental abnormalities. Neonatal seizures often are subtle, not associated with observable clinical expression, and associated with adverse development. Electroencephalographic confirmation is important in the evaluation of neonatal seizures.


Synthesis of polymeric derivatives of isoniazid: characterization and in vitro release from a water-soluble adduct with polysuccinimide.
Coupling of isoniazid with polysuccinimide afforded a water-insoluble polymeric pro-drug; by reaction with ethanolamine it was chemically transformed in a water-soluble adduct. The in vitro release of isoniazid from the drug-polymer adduct was studied by using an artificial stomach wall lipid membrane. The transfer rate constant from simulated gastric juice to simulated plasma was defined and compared with that of an equivalent dose of pure drug.


Regulatory role of proteasome in determination of platelet life span.
Limit of platelet life span (8-10 days) is determined by the activity of a putative "internal clock" composed of Bcl-2 family proteins, whereas the role of other molecular players in this process remains obscure. Here, we sought to establish a central role of proteasome in platelet life span regulation. Administration of mice with inhibitors of proteasome peptidase activity induced significant thrombocytopenia. This was associated with enhanced clearance of biotin-labeled platelets from circulation and reduction in average platelet half-life from 66 to 37 h. Cells pretreated in vitro with proteasome inhibitors exhibited augmented annexin V binding and a drop in mitochondrial transmembrane potential indicative of apoptotic cell death and decreased platelet life span. These cells were preferentially phagocytosed by monocyte-derived macrophages, thus linking proteasome activity with platelet survival. The decisive role of proteasome in this process was underscored from enhanced expression of conformationally active Bax in platelets with attenuated proteasome activity, which was consistent with pro-apoptotic phenotype of these cells. The present study establishes a critical role of proteasome in delimiting platelet life span ostensibly through constitutive elimination of the conformationally active Bax. These findings bear potential implications in clinical settings where proteasome peptidase activities are therapeutically targeted.


Bradykinin-induced translocation of protein kinases C in neuroblastoma NCB-20 cell: dependence on 1,2-diacylglycerol content and free calcium.
Bradykinin (BK)-induced production of 1,2-diacylglycerol (1,2-DG) and translocation of protein kinases C (PKCs) were examined in neuroblastoma-derived hybrid NCB-20 cells. Mass analysis of 1,2-DG exhibited a biphasic increase by 1 microM BK stimulation: the first transient phase and the second broad sustained phase. Among three subspecies of PKC expressed in these cells, types II and III were observed to translocate from cytosol to membrane in response to BK as well as PBt2 by Western blotting analysis. Type II translocated more rapidly and distinctly than type III. However, after treatment with quin 2/AM, the second phase of 1,2-DG formation completely disappeared and PKCs translocation by BK or PBt2 was completely abolished. BK-induced IP3 (1,4,5) formation was temporally consistent with the first transient phase of 1,2-DG formation. These findings suggest that PKCs translocation by BK stimulation is caused by 1,2-DG produced not only via phosphoinositide metabolism, but via other phospholipid breakdown which is Ca2+-dependent.


Principles for the Prevention of Medication-Induced Nephrotoxicity.
Medications are a common cause of injury to the kidney and can contribute to the increased progression of disease, poorer outcomes, and increased health care costs. Improved prescribing practices can decrease the risk for the development of acute kidney injury and the progression to end-stage kidney disease. KDIGO Clinical Practice Guidelines recommend the use of caution when prescribing potentially nephrotoxic medications for patients with kidney disease. More than 50-72% of individuals across all stages of kidney disease utilized potentially nephrotoxic medications contributing to poorer outcomes. Annually, 1.5 million adverse drug events causing medication-induced nephrotoxicity occur in the US. Medication-induced nephrotoxicity accounts for 14-26% of cases of AKI in adults and 16% of hospitalized children. It is imperative that nurses and all health care providers are practicing nephrotoxic stewardship to prevent medication-induced nephrotoxicity.


Transcriptional analysis of the ostA/imp gene involved in organic solvent sensitivity in Escherichia coli.
Transcriptional initiation sites of the ostA gene involved in organic solvent sensitivity in Escherichia coli were found by primer extension analysis. Two transcriptional initiation sites were newly identified at -133 and -48 nucleotides from the initiation codon of ostA, but the previously reported sigmaE-dependent one at -227 could not be detected. No heat-inducible expression of ostA was observed by Northern blotting analysis, indicating that the contribution of sigmaE-dependent transcription was very small if any. SigmaD-dependent promoter-like sequences were found just upstream of the newly identified transcriptional initiation sites by computer-aided analysis. Deletion analysis of ostA-lacZ fusions demonstrated that these two promoters contributed almost equally to the constitutive expression of the ostA gene.


Protein phosphorylation and activation of human platelets by sodium fluoride.
The ability of sodium fluoride (NaF) and thrombin to stimulate aggregation and protein phosphorylation in intact human platelets was measured and compared. When platelets were stimulated by NaF, phosphorylation of the 20 KDa protein was transient and after 5-10 min returned to the same level as that of unstimulated cells. On the other hand, 47 KDa protein was slowly phosphorylated without obvious dephosphorylation. The slow activation of the 47 KDa protein phosphorylation correlated well with the time required for the aggregation and secretion. Phosphoamino acid analysis showed that the phosphorylated amino acids of the 47 KDa protein from platelets activated by NaF and thrombin were slightly different. These results suggest that different stimuli may lead to the same protein phosphorylation by different biochemical mechanisms of action.


Hyperammonemia Syndrome After Lung Transplantation: A Double-Hit Fatal Syndrome. A Case Report.
Hyperammonemia after lung transplantation is a rare but potentially fatal condition. A 59-year-old male patient affected by pulmonary fibrosis underwent an uncomplicated bilateral lung transplant. Fourteen days after the procedure, the patient developed severe encephalopathy caused by elevated serum ammonia levels. Ureaplasma parvum and Mycoplasma hominis were found on bronchial aspirate and urinary samples as well as on pharyngeal and rectal swabs. Despite the initiation of multimodal therapy, brain damage due to hyperosmolarity was so extensive to evolve into brain death. The autopsy revealed glutamine synthetase hypo-expression in the hepatic tissue. The pathophysiology of hyperammonemia syndrome in lung transplant recipients remains unclear. Previous studies have described the presence of disorders of glutamine synthetase, while others considered the infection with urea-splitting microorganisms as a cause of hyperammonemia syndrome. Our report describes the case of a patient who developed hyperammonemia after a lung transplant in which both the aforementioned etiologies were documented. A high level of clinical suspicion for hyperammonemia syndrome should be maintained in lung transplant recipients. Timely recognition and treatment are critical to prevent the potentially dreadful evolution of this severe complication.