Hydrogen sulfide prevents formaldehyde-induced neurotoxicity to PC12 cells by attenuation of mitochondrial dysfunction and pro-apoptotic potential.
Hydrogen sulfide (H(2)S) has been shown to act as a neuroprotectant and antioxidant. Numerous studies have demonstrated that exposure to formaldehyde (FA) causes neuronal damage and that oxidative stress is one of the most critical effects of FA exposure. Accumulation of FA is involved in the pathogenesis of Alzheimer's disease (AD). The aim of present study is to explore the inhibitory effects of H(2)S on FA-induced cytotoxicity and apoptosis and the molecular mechanisms underlying in PC12 cells. We show that sodium hydrosulfide (NaHS), a H(2)S donor, protects PC12 cells against FA-mediated cytotoxicity and apoptosis and that NaHS preserves the function of mitochondria by preventing FA-induced loss of mitochondrial membrane potential and release of cytochrome c in PC12 cells. Furthermore, NaHS blocks FA-exerted accumulation of intracellular reactive oxygen species (ROS), down-regulation of Bcl-2 expression, and up-regulation of Bax expression. These results indicate that H(2)S protects neuronal cells against neurotoxicity of FA by preserving mitochondrial function through attenuation of ROS accumulation, up-regulation of Bcl-2 level, and down-regulation of Bax expression. Our study suggests a promising future of H(2)S-based preventions and therapies for neuronal damage after FA exposure.


Evidence for macrophage-mediated myelin disruption in an animal model for Charcot-Marie-Tooth neuropathy type 1A.
Charcot-Marie-Tooth neuropathy type 1A (CMT 1 A) is the most common inherited neuropathy in humans and is mostly caused by a 1.5-Mb tandem duplication of chromosome 17 comprising the gene for the peripheral myelin protein 22-kDa (PMP 22). Although there are numerous studies on the functional role of PMP 22, the mechanisms of myelin degeneration under PMP 22-overexpression conditions have not yet been fully understood. We have shown previously that in mouse mutants hetero- or homozygously deficient for two other myelin components, P0 and C x 32, respectively, immune cells contribute to the demyelinating neuropathy. To test this possibility for PMP 22 overexpression, we investigated a putative mouse model for CMT 1 A, i.e., the mouse strain C 6 1 mildly overexpressing human PMP 22 in peripheral nerves. Electron microscopic and electrophysiologic investigations revealed that this mouse strain develops pathologic features similar to those found in CMT 1 A patients. A novel finding, however, was the upregulation of CD8- and F4/80-positive lymphocytes and macrophages, respectively, in peripheral nerves. The observation that macrophages enter endoneurial tubes of the mutants and obviously phagocytose morphologically normal myelin strongly suggests that the myelin degeneration is mediated at least partially by these phagocytic cells. By gene array technology and quantitative RT-PCR of peripheral nerve homogenates from PMP 22 mutants, monocyte chemoattractant protein-1 (MCP-1; cc l2) could be identified as a putative factor to attract or activate macrophages that attack myelin sheaths in this model of CMT 1 A.


Moral distress among critical care nurses when excecuting do-not-resuscitate (DNR) orders in a public critical care unit in Gauteng.
A critical care unit admits on a daily basis patients who are critically ill or injured. The condition of these patients' may deteriorate to a point where the medical practitioner may prescribe or decide on a 'do not resuscitate' (DNR) order which must be executed by a professional nurse, leading to moral distress which may manifest as poor teamwork, depression or absenteeism. To explore and describe factors contributing to moral distress of critical care nurses executing DNR orders. The explorative descriptive qualitative design was selected to answer the research questions posed. Critical care nurses of a selected public hospital in Gauteng Province were selected via purposive sampling to participate in the study, and data were collected through semi-structured interviews. A shift leader assisted with selection of participants who met the eligibility criteria. The mean age of the participants was 36 years; most of them had more than five years' critical care nursing experience. Twelve critical care nurses were interviewed when data saturation was reached. Thereafter two more interviews were conducted to confirm data saturation. A total of 14 interviews were conducted. Tesch's eight-step method was utilised for data analysis. The findings were classified under three main themes: moral distress, communication of DNR orders and unavailability of psychological support for nurses. The findings revealed that execution of DNR orders is a contributory factor for moral distress in critical care nurses. National guidelines and/or legal frameworks are required to regulate processes pertaining to the execution of DNR orders. The study further demonstrated the need for unit-based ethical platforms and debriefing sessions for critical care nurses. The main contribution of this study was to explore and describe the factors contributing to Moral distress when executing a DNR order. This study raised awareness amongst healthcare providers on the factors contributing to moral distress amongst critical care nurses. This study highlighted the importance of developing national guidelines and legal frameworks pertaining to execution of DNR orders. This study alluded to the value of initiating debriefing sessions for critical care nurses involved in the execution of DNR orders.


Effectiveness and Safety of Direct Oral Anticoagulants in an Asian Population with Atrial Fibrillation Undergoing Dialysis: A Population-Based Cohort Study and Meta-Analysis.
Whether direct oral anticoagulants (DOACs) are more effective and safer than warfarin among Asian patients with non-valvular atrial fibrillation (NVAF) undergoing dialysis remains unclear. We first compared the risks of ischemic stroke/systemic embolism (IS/SE) and major bleeding associated with DOACs compared with warfarin, in NVAF Asians undergoing dialysis using the Taiwan National Health Insurance Research Database (NHIRD) (Aim 1). Next, we searched PubMed and Medline from January 1, 2010 until January 31, 2020, to perform a systematic review and meta-analysis of all observational real-world studies comparing DOACs with warfarin specifically focused on NVAF patients with stage 4 or 5 chronic kidney disease undergoing dialysis (Aim 2). Finally, we tested the hypothesis whether AF patients undergoing dialysis treated with OACs (warfarin and DOACs) would be associated with lower risk of adverse clinical outcomes as compared to those without OACs using the Taiwan NHIRD (Aim 3). From June 1, 2012, to December 31, 2017, a total of 3237 and 9263 NVAF patients comorbid with ESRD receiving oral anticoagulant (OACs) (490 on DOAC, 2747 on warfarin) or no OACs, respectively, were enrolled. Propensity score matching was used to balance covariates across the study groups. For the comparison of DOAC vs. warfarin (Aim 1), DOACs had comparable risks of IS/SE and major bleeding to warfarin in our present cohort. From the original 85 results retrieved, nine studies (including our study) with a total of 6490 and 22,494 patients treated with DOACs and warfarin were included in the meta-analysis, respectively. There were 5343 (82%) and 20,337 (90%) patients treated with DOACs and warfarin undergoing dialysis, respectively. The pooled meta-analysis also indicated no difference of the effectiveness (HR:0.90; [95%CI:0.74-1.10]; P = 0.32) and safety outcomes (HR:0.75; [95%CI:0.54-1.05]; P = 0.09) between DOACs and warfarin (Aim 2). For the comparison of OAC (+) vs. OAC (-) (Aim 3), OAC-treatment was associated with a higher risk of IS/SE (hazard ratio (HR):1.54; [95% confidential interval (CI):1.29-1.84];P < 0.0001) and comparable risk of major bleeding compared to those without OAC treatment. DOACs did not provide benefit over warfarin regarding effectiveness and safety in AF patients undergoing dialysis. The use of OAC was not associated with a lower risk of IS/SE in ESRD AF patients when compared to those without OAC use.


The prophylactic use of cyclooxygenase inhibitors in recurrent herpes simplex infections.
In an open study, continuous oral therapy with cyclooxygenase inhibitors (ibuprofen or indomethacin) reduced the incidence and frequency of recurrent eruptions in nine of 16 patients with herpes simplex virus infection. Retrospective analysis of seven cases indicated that natural killer cell enhancement assays might allow us to predict the clinical outcome of such treatment.


Radiotherapy for primary lung cancer.
Herein are presented the recommendations from the Société française de radiothérapie oncologique regarding indications and modalities of lung cancer radiotherapy. The recommendations for delineation of the target volumes and organs at risk are detailed.


Correlated histological and morphometric study of kidney and adrenal gland from Guinea pig exposed to hyperbaric-hyperoxic environment.
Our study revealed morphological changes in the two organs of Guinea pig kidney and suprarenal gland exposed to hyperbaric-hyperoxic environment. Proceeding from these data and knowing that in hyperbaric-hyperoxic environment the production of free oxygen radicals is increased, while the afferent arterioles undergo vasoconstriction, with direct implications on the cellular metabolism, our study puts forth the survey of the influence of hyperoxic environment on kidneys and adrenal gland - complex organs which are exposed to stress.


Characterization of migration parameters on peripheral and central nervous system T cells following treatment of experimental allergic encephalomyelitis with CRYAB.
CRYAB, a small heat shock protein, was previously shown to decrease neuroinflammation in experimental allergic encephalomyelitis (EAE). We investigated whether the expression of cell adhesion molecules and chemokine receptors on peripheral and spinal cord T cells, that could possibly affect their migration to the central nervous system, was altered following EAE CRYAB treatment. Less LFA-1+ lymphocytes and lower levels of iTAC, MCP-5 and MIG were observed in spinal cords of CRYAB-injected EAE animals. In addition, fewer blood T cells expressed CCR6, CXCR4 and CCR7 and in vivo-derived CRYAB EAE CD4+ lymphocytes were less migratory towards a MIP-3alpha gradient in vitro.


Effect of neonatal treatment with capsaicin on carrageenan-induced paw oedema in the rat.
The time course of the paw oedema induced by the subplantar injection of carrageenan was studied in rats treated neonatally with capsaicin and in their vehicle-treated controls. In the capsaicin-treated rats, which show a permanent deficit of unmyelinated primary sensory neurones, carrageenan produced an oedema which was larger and lasted longer than in the vehicle-treated rats. Pretreatment with the histamine liberator compound 48/80 reduced the carrageenan-induced paw oedema only in the capsaicin-treated rats whereas pretreatment with indomethacin reduced it in both groups of rats. The increased and prolonged inflammatory response to carrageenan in capsaicin-treated rats may be explained by an enhanced release of histamine from mast cells and may also reflect a 'trophic disorder of the denervated skin'.


Uncoupling of VEGF with NO as a mechanism for diabetic nephropathy.
VEGF plays protective roles on a variety of non-diabetic renal diseases. However, in diabetes VEGF exhibits deleterious roles to mediate the development/progression of diabetic nephropathy in spite of high VEGF. The protective role of VEGF could be predominantly dependent on its ability to stimulate nitric oxide production in endothelial cell in non-diabetic renal disease. However, it has been known that nitric oxide bioavailability is reduced in diabetes, indicating that diabetic status does not allow high VEGF to lead to an increase in NO bioavailability. As a result, VEGF could engage to NO-independent pathway, and cause deleterious effects on vascular system. Thus, we have hypothesized that uncoupling of VEGF with endothelial NO can be a mechanism by which VEGF causes diabetic nephropathy. We found that diabetic eNOS knockout (KO) mice exhibit masangiolysis, glomerular capillary microaneurysm, Kimmelstiel-Wilson-like nodular lesions, abnormal angiogenesis and a marked macrophage infiltration in addition to mesangial expansion and thickening of GBM, all of that resemble human diabetic nephropathy. Interestingly these lesions were associated with an increase in renal VEGF expression, suggesting uncoupling of VEGF with endothelial NO could be a mechanism. Compatibly, our in vitro experiments demonstrated that VEGF-induced endothelial cell proliferation was enhanced by NO blocking (with LNAME) and suppressed by exogenous NO administration whereas macrophage migration in response to VEGF was inhibited by exogenous NO, suggesting that uncoupling condition could cause abnormal angiogenesis and macrophage infiltration.


Cladistic analysis of anuran POMC sequences.
Procedures for performing cladistic analyses can provide powerful tools for understanding the evolution of neuropeptide and polypeptide hormone coding genes. These analyses can be done on either amino acid data sets or nucleotide data sets and can utilize several different algorithms that are dependent on distinct sets of operating assumptions and constraints. In some cases, the results of these analyses can be used to gauge phylogenetic relationships between taxa. Selecting the proper cladistic analysis strategy is dependent on the taxonomic level of analysis and the rate of evolution within the orthologous genes being evaluated. For example, previous studies have shown that the amino acid sequence of proopiomelanocortin (POMC), the common precursor for the melanocortins and beta-endorphin, can be used to resolve phylogenetic relationships at the class and order level. This study tested the hypothesis that POMC sequences could be used to resolve phylogenetic relationships at the family taxonomic level. Cladistic analyses were performed on amphibian POMC sequences characterized from the marine toad, Bufo marinus (family Bufonidae; this study), the spadefoot toad, Spea multiplicatus (family Pelobatidae), the African clawed frog, Xenopus laevis (family Pipidae) and the laughing frog, Rana ridibunda (family Ranidae). In these analyses the sequence of Australian lungfish POMC was used as the outgroup. The analyses were done at the amino acid level using the maximum parsimony algorithm and at the nucleotide level using the maximum likelihood algorithm. For the anuran POMC genes, analysis at the nucleotide level using the maximum likelihood algorithm generated a cladogram with higher bootstrap values than the maximum parsimony analysis of the POMC amino acid data set. For anuran POMC sequences, analysis of nucleotide sequences using the maximum likelihood algorithm would appear to be the preferred strategy for resolving phylogenetic relationships at the family taxonomic level.


Querying Event Sequences by Exact Match or Similarity Search: Design and Empirical Evaluation.
Specifying event sequence queries is challenging even for skilled computer professionals familiar with SQL. Most graphical user interfaces for database search use an exact match approach, which is often effective, but near misses may also be of interest. We describe a new similarity search interface, in which users specify a query by simply placing events on a blank timeline and retrieve a similarity-ranked list of results. Behind this user interface is a new similarity measure for event sequences which the users can customize by four decision criteria, enabling them to adjust the impact of missing, extra, or swapped events or the impact of time shifts. We describe a use case with Electronic Health Records based on our ongoing collaboration with hospital physicians. A controlled experiment with 18 participants compared exact match and similarity search interfaces. We report on the advantages and disadvantages of each interface and suggest a hybrid interface combining the best of both.


Presence of renin in primary neuronal and glial cells from rat brain.
Immunocytochemical and biochemical techniques have been utilized in the present study to characterize renin in brain cell cultures. With the use of renin-specific antibody, positive renin staining was seen in neuronal and in astrocytic glial cells using the peroxidase-antiperoxidase method. Renin concentration was pH-dependent with highest concentrations at 5.5, decreasing from pH 6.0 to 6.5. At pH 7.4 no renin was detectable in either glial or neuronal cells. The contribution of cathepsin D to the measured renin was about 10% at pH 5.5; 7% at pH 6.0 and 3% at pH 6.5. Comparison of glial with neuronal cells from WKY rats revealed significantly elevated renin at pH 5.5 in glial cells. No difference was seen between glial and neuronal renin levels in WKY rats at pH 6.0 and 6.5. At pH 5.5 and 6.0 renin was significantly increased in neuronal cells of SHR compared to WKY, whereas at pH 6.5 no difference was observed. The renin concentration in cells kept for 2 days in serum-free medium did not differ from those measured in cells kept in serum-containing medium. The generated peptide was identified as [Ile5]Angiotensin I on reversed-phase HPLC.


Global, regional and national availability of essential medicines for children, 2009-2020: a systematic review and meta-analysis.
Access to essential medicines is a vital component of universal health coverage. The low availability of essential medicines for children (EMC) has led the World Health Organization (WHO) to issue a number of resolutions calling on member states on its improvement. But its global progress has been unclear. We aimed to systematically evaluate the progress of availability of EMC over the past decade across economic regions and countries. We searched eight databases from inception to December 2021 and reference lists to identify included studies. Two reviewers independently conducted literature screening, data extraction and quality evaluation. This study was registered with PROSPERO, CRD42022314003. Overall, 22 cross-sectional studies covering 17 countries, 4 income groups were included. Globally, the average availability rates of EMC were 39.0% (95%CI: 35.5-42.5%) in 2009-2015 and 43.1% (95%CI: 40.1-46.2%) in 2016-2020. Based on the World Bank classification of economic regions, income was not proportional to availability. Nationally, the availability rate of EMC was reasonable and high (> 50%) in only 4 countries, and low or very low for the rest 13 countries. The availability rates of EMC in primary healthcare centers had increased, while that for other levels of hospitals slightly declined. The availability of original medicines decreased while that of generic medicines was stable. All drug categories had not achieved the high availability rate. The availability rate of EMC was low globally, with slight increase in the last decade. Continuous monitoring and timely reporting of the availability of EMC are also needed to facilitate targets setting and inform relevant policy making.


Preparative density gradient centrifugation of RNA and DNA in cesium sulfate-urea mixture.
A new procedure is described for a large scale separation and purification of unfixed DNA and RNA from a mixture of partially extracted nucleic acids and lysates of subcellular fractions by centrifugation to equilibrium in cesium sulfate-urea mixture. Optimum conditions are described for the separation and quantative recovery of both RNA and DNA in a pure form. The procedure allows determination of peak buoyant densities of 4-5s RNA, 7-11s mRNA and total cytoplasmic RNA. The procedure also allows fractionation of small molecular weight classes of cytoplasmic RNAs from the 18s and 28s rRNAs.


Prenatal diagnosis of citrullinemia type 1: a Chinese family with a novel mutation of the ASS1 gene.
Argininosuccinate synthetase deficiency (citrullinemia type 1) is a rare autosomal recessive disorder of the urea cycle characterized by elevated concentrations of citrulline, ammonia, and orotic acid, manifesting with acute hyperammonemic crises, usually early in life, with concurrent neurologic deterioration. Only a few cases of citrullinemia type 1 have been documented from mainland China. Prenatal diagnosis has not been performed. A Chinese family affected by citrullinemia type 1 was studied. The proband, a girl, was the second child born to a non-consanguineous couple. Her elder brother died at 19months due to coma and liver dysfunction of unknown cause. The proband was admitted because of severe mental retardation and lethargy at the age of 15months. Initial laboratory results revealed hyperammonaemia, hypercitrullinemia (928.771μmol/L, normal 5.0-25.0μmol/L) and orotic aciduria, supporting the diagnosis of citrullinemia type 1. Subsequently, the mother presented at 15weeks of pregnancy seeking for genetic counseling and prenatal diagnosis. ASS1 gene in the blood leukocytes of the family members and amniocytes was performed by direct sequencing. On the ASS1 gene of the proband, a novel mutation, T1009C (C337R), and a previously reported mutation G847A (E283K) were identified. Each parent carries one of two mutations. G847A and T1009C mutations were detected in amniocytes, as same as the proband of the family. The result revealed that the fetus was affected by argininosuccinate synthetase deficiency. The parents chose to have a termination of the pregnancy. Prenatal diagnosis for citrullinemia type 1 was performed in a Chinese family using gene analysis. T1009C (C337R), a novel mutation of ASS1, was identified.


Prediction of mixture toxicity with its total hydrophobicity.
Based on the C18 Empore disk/water partition coefficient of a mixture, quantitative structure-activity relationships (QSARs) are presented, which are used to predict the toxicity of mixed halogenated benzenes to P. phosphoreum. The predicted toxicity of 10 other related mixtures based on the QSAR model, agree well with the observed data with r2 = 0.973, SE = 0.113 and F = 287.785 at a level of significance P < 0.0001. The joint effect of these chemicals is simple similar action and the toxicity of the mixtures can be predicted from total hydrophobicity and is independent of hydrophobicity of the components or the ratio of the individual chemicals.


Microdissection, mRNA amplification and microarray: a study of pleural mesothelial and malignant mesothelioma cells.
The studies of molecular alterations in tumor cells with microarrays are often hampered by inherent tissue heterogeneity. The emergence of Laser Capture Microdissection (LCM) allowed us to overcome this challenge since it gives selective access to cancer cells that are isolated from their native tissue environment. In this report, we microdissected mesothelial cells and malignant mesothelioma cells of ex vivo resected specimens using LCM. Amplified RNA from mesothelial and mesothelioma microdissected cells allowed us to measure global gene expression with 10 K-microarrays in four independent experiments. We screened 9850 annotated human genes, 1275 of which have satisfied our data analysis requirements. They included 302 overexpressed genes and 160 downregulated genes in mesothelioma microdissected cells as compared to mesothelial microdissected cells. Among them, the expression levels of eight genes, namely BF, FTL, IGFBP7, RARRES1, RARRES2, RBP1, SAT, and TXN according to HUGO nomenclature, were increased, whereas six: ALOX5AP, CLNS1A, EIF4A2, ELK3, REQ and SYPL, were found to be underexpressed in mesothelioma microdissected cells. The ferritin light polypeptide (FTL) gene overexpression was confirmed by real time quantitative PCR. Our approach allowed a comprehensive in situ examination of mesothelioma and provided an accurate way to find new marker genes that may be useful for diagnosis and treatment of malignant pleural mesothelioma.


Genetic analysis of two Taiwanese bluetongue viruses.
BTV2/KM/2003 and BTV12/PT/2003 are the first identified bluetongue viruses in Taiwan. The prototype virus BTV2/KM/2003 was previously characterized in various respects as low virulent. In the present study, nucleotide sequences of the ten genome segments and their coding regions of the Taiwan strains were determined and analyzed. The two strains had >96.8% nucleotide and >97.9% deduced amino acid identities to each other, except for the VP2 genes. Their genome sequences, except for NS1 and VP2 genes, clustered overall in the Asian lineage, and were closely related to strains from China, India, Indonesia, and Japan. The phylogenetic trees and nucleotide identities of six BTV genes were suggestive of the geographical origin of the bluetongue virus strains analyzed, with a few exceptions. To examine which genes better distinguished strains from different origins (topography), the distribution of and the levels of differences in nucleotide identities were analyzed, revealing that VP3, NS2, and NS3 genes were more suitable for topotyping of BTVs. Analysis of ratios of non-synonymous/synonymous substitutions (dN/dS values) between putative ancestry and their descendant strains suggested that most BTV genes evolved under a negative selection, whereas the VP7 gene evolved under positive selection, and its non-synonymous substitutions accumulated more rapidly in strains from the Mediterranean region.


Phytochrome A regulates the intracellular distribution of phototropin 1-green fluorescent protein in Arabidopsis thaliana.
It has been known for decades that red light pretreatment has complex effects on subsequent phototropic sensitivity of etiolated seedlings. Here, we demonstrate that brief pulses of red light given 2 h prior to phototropic induction by low fluence rates of blue light prevent the blue light-induced loss of green fluorescent protein-tagged phototropin 1 (PHOT1-GFP) from the plasma membrane of cortical cells of transgenic seedlings of Arabidopsis thaliana expressing PHOT1-GFP in a phot1-5 null mutant background. This red light effect is mediated by phytochrome A and requires approximately 2 h in the dark at room temperature to go to completion. It is fully far red reversible and shows escape from photoreversibility following 30 min of subsequent darkness. Red light-induced inhibition of blue light-inducible changes in the subcellular distribution of PHOT1-GFP is only observed in rapidly elongating regions of the hypocotyl. It is absent in hook tissues and in mature cells below the elongation zone. We hypothesize that red light-induced retention of the PHOT1-GFP on the plasma membrane may account for the red light-induced increase in phototropic sensitivity to low fluence rates of blue light.


Prognostic Value of the Number of Retrieved Lymph Nodes in Pathological Nx or N0 Classical Papillary Thyroid Carcinoma.
This study evaluated the clinical implications of the number of retrieved central lymph nodes (LN) for the recurrence and recurrence-free survival (RFS) outcomes in patients with pathological Nx (pNx) or N0 classical papillary thyroid carcinoma (PTC). In total, 464 patients were enrolled following total thyroidectomy with central LN dissection. The risk factors related to recurrence and RFS were evaluated and compared between these groups. Age, primary tumor size, and number of retrieved central LNs were independent risk factors for recurrence according to multivariate analysis (p < 0.05). The cut-off value for the number of retrieved central LNs related to recurrence was 4.5. Group 2 (pN0; ≥5 nodes) demonstrated a significantly higher proportion of patients with an ablation-stimulated thyroglobulin (sTg) level <2.0 ng/mL (84.9 vs 61.1 %; p < 0.050) and control sTg level <1.0 ng/mL (92.1 vs 79.6 %; p < 0.050) in comparison with patients in group 1 (pNx or pN0; 1-4 nodes). Perioperative complication rates were comparable between groups. The number of retrieved central LNs is an independent risk factor for recurrence, even among patients with pNx or pN0 classical PTC. A thorough central LN dissection may therefore improve the long-term RFS rate.


Amplification of chloramphenicol resistance transposons carried by phage P1Cm in Escherichia coli.
We have characterized a number of P1Cm phages which contain the resistance genes to chloramphenicol and fusidic acid as IS1-flanked Cm transposons. Restriction cleavage and electron microscopic analysis showed that these Cm transposons were carried as monomers (M) or tandem dimers (D). Lysogens of P1Cm (D) are more resistant to chloramphenicol than those of its P1Cm (M) presumably as a result of an increased gene dosage. Amplification of the Cm transposons to tandem multimers was frequently observed in P1Cm (D) lysogens grown in the presence of high concentrations of chloramphenicol or fusidic acid and was also detected in P1Cm (M) lysogens. The degree of amplification varied in different clones which suggests that cells containing spontaneously amplified Cm transposons were selected by high doses of the antibiotics. The dimeric as well as the amplified Cm transposons carried in P1Cm lysogens grown in the absence of chloramphenicol displayed considerable stability. Mechanisms for the amplification of the IS1-flanked transposons are discussed.


Quantitative determination of propildazine in rat plasma by gas-liquid chromatography.
A gas-liquid chromatographic method for the evaluation of the new anti-hypertensive drug propildazine (ISF 2123) in rat plasma is described. The procedure involves separation of the drug from plasma by cation-exchange chromatography, subsequent acylation of the dried eluate with heptafluorobutyric anhydride and quantitation with electron-capture detection. Propildazine can be determined in concentrations down to ca. 0.4 microgram/ml.


Secondary immunoglobulin A nephropathy with gross hematuria leading to rapidly progressive glomerulonephritis following severe acute respiratory syndrome coronavirus 2 vaccination: a case report.
The outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has been followed by many reports of the development and relapse of autoimmune diseases associated with SARS-CoV-2 vaccination. Some of these reports have involved relapse or onset of immunoglobulin A (IgA) nephropathy following SARS-CoV-2 vaccination. Here, we report on a patient with IgA nephropathy who presented with gross hematuria and rapidly progressive glomerulonephritis following SARS-CoV-2 vaccination. A 63-year-old male patient with a history of habitual tonsillitis underwent bilateral tonsillectomy. He had a history of alcoholic cirrhosis of the liver and microscopic hematuria and proteinuria were indicated during a health checkup 2 years before hospital admission. He developed hematuria after the SARS-CoV-2 vaccination, which led to rapidly progressive glomerulonephritis, for which he was hospitalized. A renal biopsy led to the diagnosis of IgA nephropathy. Although pulse steroid therapy during his condition resulted in hepatic encephalopathy, three courses combined with mizoribine improved his renal function. SARS-CoV-2 mRNA vaccines activate T cells, which are involved in the pathophysiology of IgA nephropathy. Therefore, this case suggests that the exacerbation of IgA nephropathy by the vaccine favors the vasculitis aspect of the disease.


Experience of Comamonas acidovorans keratitis with delayed onset and treatment response in immunocompromised cornea.
To report 2 cases of Comamonas acidovorans keratitis in immunocompromised cornea. A complete review of the medical records of the two cases of Comamonas acidovorans keratitis. We found some similarities in clinical courses of two cases. Both of them showed development of keratitis during the management with corticosteroids, delayed onset, slow response to antibiotics, and relatively less affected corneal epithelium. Comamonas acidovorans is known as a less virulent organism. However it can cause an indolent infection that responds slowly even to adequate antibiotics therapy in immunocompromised corneas.


Expression of GnIH and its effects on follicle development and steroidogenesis in quail ovaries under different photoperiods.
Photoperiod is an important environmental factor that influence seasonal reproduction behavior in bird and GnIH can play a function in this process through the reproductive axis, and some studies suggest that GnIH may have a direct role at the gonadal level. To investigate the expression of GnIH and its effects on follicle development and steroidogenesis in quail ovaries under different photoperiods, 72 healthy laying quails of 8-wk-old were randomly divided into long day (LD) group [16 light (L): 8 dark (D)] (n = 36) and short day (SD) group (8L:16D) (n = 36). Samples were collected from each group on d1, d11, d22, and d36 of the experiment. The result showed that short day treatment upregulated the level of GnIH in the gonads (P < 0.05), decreased the expression level of CYP19A1,3β-HSD, StAR, LHR, and FSHR and increased the expression level of AMH, AMHR2, GDF9, and BMP15 to inhibit follicle development and ovulation, thus affecting the egg production performance of quails. In vitro culture of quail granulosa cells and treatment with different concentrations of GnIH (0, 1, 10, and 100 ng/mL) for 24 h. Result showed that GnIH inhibited the levels of FSHR, LHR, and steroid synthesis pathways in granulosa cells, upregulated the levels of AMHR2, GDF9, and BMP15. The results suggest that the inhibition of follicle development and reduced egg production in quail by short day treatment is due to GnIH acting at the gonadal level, and GnIH affected the steroid synthesis by inhibiting gonadotropin receptors.


Arrhythmogenic effects of endothelin-1 under conditions of NO-synthase blockade with L-NAME in NMRI mice.
Arrythmogenic effects of endothelin-1 were studied in NMRI mice under conditions of NO-synthase blockade with N omega-nitro-L-arginine methyl ester. Intravenous injection of endothelin-1 increased heart rate variability in awake mice. NO-synthase blockade potentiated the arrythmogenic effects of endothelin-1. In narcotized animals the arrythmogenic effect of endothelin-1 was not observed and was considerably weakened under conditions of NO-synthase blockade. Arrhythmia was paralleled by atrioventricular block and lengthening of the ST segment.


Central domain of the human cardiac muscle ryanodine receptor does not mediate interaction with FKBP12.6.
The immunophilin, FK506-binding protein (FKBP12), is an essential component of the ryanodine receptor channel complex of skeletal muscle (RyR1) and modulates intracellular calcium signaling from the endoplasmic reticulum. The cardiac muscle RyR isoform (RyR2) specifically associates with a distinct FKBP isoform, FKBP12.6. Previous studies have led to the proposal that the central domain of RyR1 exclusively mediates the interaction with FKBP12. To characterize the topography of the FKBP12.6 binding site on the human cardiac RyR2, we have applied complementary protein-protein interaction methods using both in vivoyeast two-hybrid analysis and in vitroimmunoprecipitation experiments. Our results indicate an absence of interaction of FKBP12/12.6 with fragments containing the central domain of either RyR1, RyR2, or RyR3. Furthermore, no interaction was detected between FKBP12.6 with a series of overlapping fragments encompassing the entire RyR2, either individually or in multiple combination. We also found that a distinct, alternatively spliced variant of FKBP12.6 was unable to interact with RyR. In contrast, we successfully demonstrated a robust association between the cytoplasmic domain of transforming growth factor-beta receptor type I and both FKBP12 and FKBP12.6 in parallel positive control experiments, as well as between native RyR2 and FKBP12.6. These results suggest that the specific interaction of FKBP12.6 with RyR2, and generally of FKBPs with any RyR isoform, is not readily reconstituted by peptide fragments corresponding to central RyR domains. Further structural analysis will be necessary to unravel this intricate signaling system and the current model of FKBP12-RyR interaction via a single, central RyR epitope may therefore require revision.


Conservative laparoscopy in the treatment of posttraumatic splenic laceration using microfiber hemostatic collagen: three case histories.
Three cases of posttraumatic splenic laceration in young patients are reported. In each case, hemostasis of the lesion was obtained by the use of microfiber collagen in emergency laparoscopy. In two cases, complete hemostasis was achieved and the entire organ was saved. In the third case, splenectomy proved necessary after renewed bleeding occurred.


Immunohistochemical Markers as Predictors of Histopathologic Response and Prognosis in Rectal Cancer Treated with Preoperative Adjuvant Therapy: State of the Art.
We explain the state of the art of the immunohistochemical markers of response in rectal cancers treated with neoadjuvant medical therapies and its implication with prognosis. Neoadjuvant chemoradiotherapy is widely used to improve the outcome of patients with locally advanced rectal cancer, and the evaluation of the effects of medical therapy is to date based on histomorphological examination by applying four grading systems of response to therapy (tumor regression grade (TRG)). The need to identify immunohistochemical markers that could ensure a better assessment of response and possibly provide additional prognostic information has emerged. We identified p53, p27kip1, Ki67, matrix metalloprotease-9, survivin, Ki67 proliferative index, CD133, COX2, CD44v6, thymidylate synthase, thymidine phosphorylase, and dihydropyrimidine dehydrogenase as the most common markers studied in literature to date, and we explained their prognostic potential and their implications in the evaluation of the response to preoperative therapies in rectal cancers.


Development of massive multilevel molecular dynamics simulation program, Platypus (PLATform for dYnamic Protein Unified Simulation), for the elucidation of protein functions.
A massively parallel program for quantum mechanical-molecular mechanical (QM/MM) molecular dynamics simulation, called Platypus (PLATform for dYnamic Protein Unified Simulation), was developed to elucidate protein functions. The speedup and the parallelization ratio of Platypus in the QM and QM/MM calculations were assessed for a bacteriochlorophyll dimer in the photosynthetic reaction center (DIMER) on the K computer, a massively parallel computer achieving 10 PetaFLOPs with 705,024 cores. Platypus exhibited the increase in speedup up to 20,000 core processors at the HF/cc-pVDZ and B3LYP/cc-pVDZ, and up to 10,000 core processors by the CASCI(16,16)/6-31G** calculations. We also performed excited QM/MM-MD simulations on the chromophore of Sirius (SIRIUS) in water. Sirius is a pH-insensitive and photo-stable ultramarine fluorescent protein. Platypus accelerated on-the-fly excited-state QM/MM-MD simulations for SIRIUS in water, using over 4000 core processors. In addition, it also succeeded in 50-ps (200,000-step) on-the-fly excited-state QM/MM-MD simulations for the SIRIUS in water.


[National registry on cardiac electrophysiology (2010 and 2011)].
Based on a survey sent to Portuguese centers that perform diagnostic and interventional electrophysiology and/or implant cardioverter-defibrillators (ICDs), the authors analyze the number and type of procedures performed during 2010 and 2011 and compare these data with previous years. In 2011, a total of 2533 diagnostic electrophysiologic procedures were performed, which were followed by ablation in 2013 cases, a steady increase over previous years. The largest share of this increase compared to 2010 was in atrial fibrillation, which is now the second most frequent indication for ablation, after atrioventricular nodal reentrant tachycardia. The total number of ICDs implanted in 2011 was 1084, of which 339 were biventricular (BiV) cardiac resynchronization devices (BiV ICDs). This represents an increase in the total number relative to previous years, 2011 being the first year in which the rate of new ICD implantations in Portugal exceeded 100 per million population. However, compared to 2010, the number of BiV ICDs implanted decreased, despite the recent publication of updated European guidelines on device therapy in heart failure, which clarified and expanded the indications for implantation of these devices. Some comments are made on the current status of cardiac electrophysiology in Portugal and on factors that may influence its development in the coming years.


Small intestine and ovarian metastasis in a patient with a history of cardiac fibrosacoma.
Metastatic tumors involving the small bowel are much more common than primary neoplasms. The most common metastases to the small intestine are those arising from other intra-abdominal organs. Metastases from extra-abdominal tumors are rare but may be found in patients with adenocarcinoma of the breast and carcinoma of the lung. Cutaneous melanoma is the most common extra-abdominal source involving the small intestine, with involvement of the small intestine noted in more than half of the patients dying from malignant melanoma [1]. While intestinal metastasis from sarcoma has been described, this is an extremely rare occurrence especially from a rare malignant sarcoma of cardiac origin. The dismal prognosis of cardiac sarcomas results from extensive local invasion at presentation or distant metastasis. Metastasis to the small bowel may cause obstruction, bleeding, or intussusception in which the diagnosis may be delayed because of rarity of the condition and mild and vague abdominal symptoms at early presentation. In this report, a 35 year old woman a known case of cardiac fibrosarcoma was admitted to the emergency ward with abdominal pain and distention, bloody diarrhea, and recurrent nausea and vomiting. Jejuno-jejunal invagination was diagnosed at laparotomy along with tumoral involvement of the left ovary. Histopathological study showed that there was a fibrosarcoma compatible with the earlier diagnosis of primary cardiac tumor. We have described some aspects of diagnosis and treatment of this rare cause of intestinal intussusception.


Rapid turnover of actin in dendritic spines and its regulation by activity.
Dendritic spines are motile structures that contain high concentrations of filamentous actin. Using hippocampal neurons expressing fluorescent actin and the method of fluorescence recovery after photobleaching, we found that 85 +/- 2% of actin in the spine was dynamic, with a turnover time of 44.2 +/- 4.0 s. The rapid turnover is not compatible with current models invoking a large population of stable filaments and static coupling of filaments to postsynaptic components. Low-frequency stimulation known to induce long-term depression in these neurons stabilized nearly half the dynamic actin in the spine. This effect depended on the activation of N-methyl-D-aspartate (NMDA) receptors and the influx of calcium. In neurons from mice lacking gelsolin, a calcium-dependent actin-binding protein, activity-dependent stabilization of actin was impaired. Our studies provide new information on the kinetics of actin turnover in spines, its regulation by neural activity and the mechanisms involved in this regulation.


Gabapentin reverses mechanical allodynia induced by sciatic nerve ischemia and formalin-induced nociception in mice.
The anticonvulsant drug gabapentin has been demonstrated to alleviate symptoms of painful diabetic neuropathy as well as other types of neuropathic pain. The aim of the present study was to investigate the effect of gabapentin in a recently developed mouse model of peripheral neuropathy. This model is based on a photochemical ischemic lesion of the sciatic nerve generated by laser-induced activation of the photosensitizing dye erythrosin B. Following laser irradiation of the sciatic nerve for 2, 5, or 10 min, tactile allodynia was observed during at least 3 weeks. The degree of allodynia was most marked following 10 min of irradiation. Subcutaneous administration of gabapentin [175-300 micromol/kg ( approximately 30-51 mg/kg), cumulative doses, at 1-h intervals] significantly reversed tactile allodynia induced by 10-min laser irradiation. The maximal dose of gabapentin increased the withdrawal threshold from approximately 0.55 to approximately 1.85 g (i.e., about 77% of the threshold in normal animals, approximately 2.4 g). Gabapentin did not affect the tactile withdrawal threshold in intact animals. A dose of gabapentin (100 micromol/kg, sc) that had no effect on allodynia was found to significantly reduce the pain behavior during phase 2 of the formalin test. The present study demonstrates that systemic administration of gabapentin suppresses both allodynia induced by an ischemic lesion of the sciatic nerve and pain behavior in the formalin test.


Occurrences and removal of pharmaceutical and personal care products from aquatic systems using advanced treatment- A review.
Pharmaceuticals, personal care items, steroid hormones, and agrochemicals are among the synthetic and indigenous products that make up micropollutants, also known as emerging contaminants. Pharmaceutical and personal care products (PPPs) are a class of developing micropollutants that can harm living organisms even at low concentrations. Many are detected in surface water and wastewater from the treatment process, with quantities ranging from ng L-1 to gL-1; however, residual PPPs at dangerously high levels have indeed recently been recognized in the ecosystem. Residential sewage treatment plant (STP) dump the largest majority of these pollutants into the environment on a regular basis. As a result of its robust structure, it has a longer lifespan in the environment. This review article discusses how surface water pollutants such pesticides, petroleum hydrocarbons, and perfluorinated compounds affect water quality, as well as the most cost-effective adsorbents for removing these PPPs. The goal of this study is to provide information about the origins of PPP, as well as diagnostic procedures and treatment options. Research on developing contaminants is also aimed at evaluating the efficacy and affordability of adsorption.


Deterministic secure quantum communication using a single d-level system.
Deterministic secure quantum communication (DSQC) can transmit secret messages between two parties without first generating a shared secret key. Compared with quantum key distribution (QKD), DSQC avoids the waste of qubits arising from basis reconciliation and thus reaches higher efficiency. In this paper, based on data block transmission and order rearrangement technologies, we propose a DSQC protocol. It utilizes a set of single d-level systems as message carriers, which are used to directly encode the secret message in one communication process. Theoretical analysis shows that these employed technologies guarantee the security, and the use of a higher dimensional quantum system makes our protocol achieve higher security and efficiency. Since only quantum memory is required for implementation, our protocol is feasible with current technologies. Furthermore, Trojan horse attack (THA) is taken into account in our protocol. We give a THA model and show that THA significantly increases the multi-photon rate and can thus be detected.


Modulation of the glycine response by Ca2+-permeable AMPA receptors in rat spinal neurones.
1. In acutely isolated rat sacral dorsal commisural nucleus (SDCN) neurones, application of kainate (KA) reversibly potentiated glycine-evoked Cl- currents (IGly) in a concentration-dependent manner. 2. The cellular events underlying the interaction between non-NMDA receptors and glycine receptors were studied by using nystatin-perforated patch and cell-attached single-channel recording modes. 3. The action of KA was not accompanied by a shift in the reversal potential for IGly. In dose-response curves, KA potentiated IGly without significantly changing glycine binding affinity. 4. GYKI 52466 blocked while NS-102 had no effect on the KA-induced potentiation of IGly. 5. The potentiation was reduced when KA was applied in a Ca2+-free extracellular solution or in the presence of BAPTA AM, and was independent of the activation of voltage-dependent Ca2+ channels. 6. Pretreatment with KN-62, a selective Ca2+-calmodulin-dependent protein kinase II (CaMKII) inhibitor, abolished the action of KA. Inhibition of calcineurin converted the KA-induced potentiation to a sustained one. 7. Single-channel recordings revealed that KA decreased the mean closing time of glycine-gated single-channel activity, resulting in an increase in the probability of channel opening. 8. It is proposed that Ca2+ entry through AMPA receptors modulates the glycine receptor function via coactivation of CaMKII and calcineurin in SDCN neurones. This interaction may provide a new postsynaptic mechanism for control of inhibitory synaptic signalling and represent one of the important regulatory mechanisms of spinal nociception.


In vivo functioning of creatine phosphokinase in human forearm muscle, studied by 31P NMR saturation transfer.
31P nuclear magnetic resonance (NMR) saturation transfer has been used to measure enzymatic flux through the creatine phosphokinase reaction in the direction of ATP synthesis in the human forearm muscle flexor digitorum superficialis. Modification of the ratio method for measurement of spin-lattice relaxation (R. Freeman, H.D.W. Hill, and R. Kaptein, J. Magn. Reson. 7, 82(1972]was tested and used to appreciably shorten the duration of the measurement. Under conditions of steady state work intracellular pH decreased slightly by 0.06 units and the spin-lattice relaxation time of phosphocreatine in muscle was unchanged, while flux from phosphocreatine to ATP was 64 +/- 10% of the resting value. This is contrary to the increase in flux of 155% predicted from previous saturation transfer studies carried out in vitro on rabbit skeletal muscle creatine phosphokinase using metabolite concentrations to mimic those in vivo (E.A. Shoubridge, J.L. Bland, and G.K. Radda, Biochim. Biophys. Acta 805, 72 (1984]. This discrepancy could be accounted for by an underestimation of the ADP concentrations to which the enzyme is exposed due to inaccurate assumptions about the total metabolite concentrations, or possibly by compartmentation of creatine phosphokinase and its reactants.


The effect of a mindful restaurant eating intervention on weight management in women.
To evaluate the effect of a Mindful Restaurant Eating intervention on weight management. Randomized control trial. Greater metropolitan area of Austin, Texas. Women (n = 35) 40-59 years old who eat out at least 3 times per week. The intervention, using 6 weekly 2-hour, small group sessions, focused on reducing calorie and fat intake when eating out through education, behavior change strategies, and mindful eating meditations. Weight, waist circumference, self-reported daily calorie and fat intake, self-reported calories and fat consumed when eating out, emotional eating, diet related self-efficacy, and barriers to weight management when eating out. General linear models examined change from baseline to final endpoint to determine differences in outcomes between the intervention and control group. Participants in the intervention group lost significantly more weight (P =.03), had lower average daily caloric (P = .002) and fat intake (P = .001), had increased diet-related self-efficacy (P = .02), and had fewer barriers to weight management when eating out (P = .001). Mindful Restaurant Eating intervention was effective in promoting weight management in perimenopausal women.


Anatomical and electrophysiological evidence for a projection from the medial preoptic area to the 'mesencephalic and subthalamic locomotor regions' in the rat.
There is considerable physiological evidence indicating that the medial preoptic area plays an important role in neural circuits mediating ingestive, thermoregulatory, and reproductive behaviors, all of which involve foraging. The current series of anatomical and electrophysiological experiments was therefore designed to characterize a direct projection from the medial preoptic area to a region in the zona incerta just dorsal to the subthalamic nucleus, which appears to lie within the 'subthalamic locomotor region', and to the pedunculopontine nucleus, which lies within the 'mesencephalic locomotor region'. First, implants of the fluorescent tracer True blue were placed in the pedunculopontine nucleus, and retrogradely labeled neurons were consistently found in dorsal regions of the medial preoptic nucleus, anteroventral preoptic nucleus, rostral tip of the medial preoptic area, lateral parts of the medial preoptic area, and median preoptic nucleus. Second, combined retrograde-immunostaining experiments indicated that a small number of galanin-stained neurons in the rostral tip of the medial preoptic area project to the pedunculopontine nucleus, whereas in nearby regions some galanin- or neurotensin-stained neurons in the lateral preoptic area, and some neurotensin-stained neurons in the substriatal gray appear to project to the pedunculopontine nucleus, as do some neurotensin- or corticotropin releasing factor (CRF)-stained cells in the bed nucleus of the stria terminalis. Third, injections of the anterograde tracer Phaseolus vulgaris leukoagglutinin (PHA-L) into various parts of the medial preoptic area all labeled axons with terminal boutons in the caudal zona incerta and pedunculopontine nucleus. Fourth, single-pulse stimuli were delivered to the zona incerta and pedunculopontine nucleus and the location of antidromically activated neurons in the medial preoptic area was mapped using extracellular recordings. Somewhat less than one-third of the cells recorded from in the medial preoptic area were antidromically activated from either site and some 14% were influenced from both sites. The application of a reciprocal collision test to a small number of neurons suggested that at least some neurons in the medial preoptic area may send collaterals to both sites. And fifth, injections of procaine into the zona incerta were shown to block the antidromic activation of medial preoptic neurons by single-pulse stimulation of the pedunculopontine nucleus.(ABSTRACT TRUNCATED AT 400 WORDS)


The effect of adding fish oil to parenteral nutrition on hepatic mononuclear cell function and survival after intraportal bacterial challenge in mice.
Parenteral nutrition (PN) is indispensable for meeting caloric and substrate needs of patients who cannot receive adequate amounts of enteral nutrition; however, PN impairs hepatic immunity. We examined the effects of ω-3 and -6 polyunsaturated fatty acids, added individually to fat-free PN, on hepatic immunity in a murine model. We focused on serum liver enzymes, cytokine production, histopathology, and the outcomes after intraportal bacterial challenge. Male Institute of Cancer Research mice were randomized into 4 groups; ad libitum chow (CHOW), fat-free PN (FF-PN), PN + fish oil (FO-PN), or PN + safflower oil (SO-PN). After the mice had been fed for 5 days, hepatic mononuclear cells (MNCs) were isolated. The number of MNCs was counted and cytokine production (tumor necrosis factor [TNF]-α and interleukin [IL]-10) by hepatic MNCs in response to lipopolysaccharide (LPS) was measured. Blood samples were analyzed for hepatobiliary biochemical parameters. Moreover, 1.0 × 10(7) pseudomonas aeruginosa were delivered by intraportal injection. Survival and histology were examined. Hepatic MNC numbers were significantly less in the FO-PN and FF-PN than in the CHOW group, whereas the SO-PN group showed moderate recovery of hepatic MNC numbers. The CHOW, FO-PN, and SO-PN groups showed LPS dose-dependent increases in TNF-α levels. These increases were blunted in the FF-PN group. IL-10 levels were increased LPS dose-dependently in the CHOW and FO-PN groups, but no marked changes were observed with LPS stimulation in the SO-PN and FF-PN groups. Plasma levels of aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase were significantly greater in the FF-PN than in the FO- and SO-PN and CHOW groups. The FO-PN group showed significantly improved survival compared with the SO-PN and FF-PN groups, showing essentially no morphologic hepatic abnormalities. Addition of fish oil to PN was advantageous in terms of reversing PN-induced deterioration of hepatic immunity, as reflected by altered cytokine production. Fish oil administration was also useful for preventing PN-induced hepatobiliary dysfunction. These changes seem to result in better survival and to protect against severe tissue damage after intraportal bacterial challenge. This therapy may have the potential to ameliorate PN-induced impairment of host immunity and thereby decrease morbidity and mortality.


T helper and B cell escape mutations within the HBc gene in patients with asymptomatic HBV infection: a study from the South-Eastern region of Iran.
Escape mutations potentially allow viruses to avoid detection and clearance by the host immune system and may represent a mechanism through which infections may persist in some patients. The association of the mutations in the HBcAg gene with Hepatitis B asymptomatic carriers (ASC) has not been studied adequately. The current study was aimed to investigate HBcAg18-27 CTL epitope mutations in ASC patients in the South-Eastern region of Iran. 100 ASC patients were selected for this study and screened for HLA-A2 using flow cytometry. HBV-DNA was extracted from the HLA-A2 positive patients and the HBc gene was amplified using PCR. Direct double sequencing was performed to analyse mutations in the HBc gene of HBV isolates from patients with ASC. Overall, 25 (25%) of individuals were HLA-A2 positive. Direct double sequencing indicated no mutations in the HBcAg18-27 epitope. However, four mutations within the T helper and three mutations within the B cell epitopes of ASC patients were identified. The lack of mutations within the HBcAg18-27 epitope suggests that the antigenicity of this region is not altered in HBV isolates of our patients and therefore antigen presentation would occur normally to the patient's immune system through HLA-A2. However, in the course of this study we revealed some novel mutations within the T helper and B cell epitopes that may affect the efficiencies of immune response of ASC patients against these novel HBV epitopes.


The Use of Deep Water Berths and the Effect of Noise on Bottlenose Dolphins in the Shannon Estuary cSAC.
The Shannon Estuary on the west coast of Ireland is one of Europe's premier deepwater berths catering for ships up to 200,000 deadweight tonnage. It is also Ireland's only designated candidate special area of conservation for bottlenose dolphins under the EU Habitats Directive. Long-term static acoustic monitoring was carried out at a number of intensive shipping sites. In 2012, noise monitoring took place over a 6-month period (at 1 site) as part of Ireland's requirements under the Marine Strategy Framework Directive (MSFD). This is the first assessment of the potential effect of vessel traffic on the behavior of this discrete dolphin population.


Cardiovascular benefits of SGLT2 inhibitors in type 2 diabetes, interaction with metformin and role of erythrocytosis: a self-controlled case series study.
Sodium-glucose cotransporter-2 inhibitors (SGLT2i) have proven cardiovascular benefits in patients with type 2 diabetes (T2D). This self-controlled case series study aims to evaluate whether metformin use and SGLT2i-associated erythrocytosis influence its cardiovascular benefits. T2D patients with metformin and/or SGLT2i prescriptions between 2015 and 2020 were identified from the Hong Kong population. Study outcomes were composite cardiovascular diseases (CVD), coronary heart disease (CHD), hospitalisation for heart failure (HHF), stroke, and erythrocytosis. Risk periods were patient-time divided into four mutually exclusive windows: (i) 'baseline period' of metformin use without SGLT2i; (ii) pre-SGLT2i period; (iii) exposure to SGLT2i without metformin; and (iv) exposure to the drug combination. Another SCCS model was applied to evaluate the association between erythrocytosis and cardiovascular outcomes regarding SGLT2i exposure. Four mutually exclusive risk periods included (i) SGLT2i exposure with erythrocytosis; (ii) SGLT2i exposure without erythrocytosis; (iii) absence of SGLT2i exposure with erythrocytosis; and (iv) absence of SGLT2i exposure without erythrocytosis. Incidence rate ratios (IRR) of events at different risk periods were estimated using conditional Poisson regression model. Among 20,861 patients with metformin and/or SGLT2i prescriptions, 2575 and 1700 patients with events of composite CVD and erythrocytosis were identified, respectively. Compared to metformin use without SGLT2i, SGLT2i initiation was associated with lower risks of composite CVD, CHD, and HHF-regardless of the presence (CVD: IRR = 0.43, 95% CI 0.37-0.51; CHD: IRR = 0.44, 95% CI 0.37-0.53; HHF: IRR = 0.29, 95% CI 0.22-0.40; all p < 0.001) and absence of concomitant metformin (CVD: IRR = 0.31, 95% CI 0.20-0.48; CHD: IRR = 0.38, 95% CI 0.25-0.59; HHF: IRR = 0.17, 95% CI 0.09-0.31; all p < 0.001); while SGLT2i was neutral on stroke risk. Compared to metformin-SGLT2i combination, exposure to SGLT2i alone was associated with comparable risks of all cardiovascular outcomes (all p > 0.05). Incidence rates of erythrocytosis at baseline, SGLT2i without and with metformin use periods were 0.75, 3.06 and 3.27 per 100 person-years, respectively. SGLT2i users who developed erythrocytosis had lower risk of HHF (IRR = 0.38, 95% CI 0.14-0.99, p = 0.049) than those who did not. Our real-world data suggested that SGLT2i-associated cardiovascular benefits were not attenuated by metformin use. Further studies will delineate the role of erythrocytosis as a surrogate marker of SGLT2i-associated cardiovascular benefit in reducing HHF.


Kaposi's sarcoma in Morocco: a pathological study with immunostaining for human herpesvirus-8 LNA-1.
The Maghreb region is considered to be an endemic area for classical Kaposi's sarcoma (KS) but the few published reports have focused on clinical descriptions. We report here the clinical, virological and pathological features of 26 KS cases, including 17 classic forms, eight epidemic forms and one case of post-transplant KS. Evidence of human herpesvirus (HHV)-8 infection was detected by serology, immunohistochemistry and polymerase chain reaction. Antibodies directed against HHV-8 latent nuclear antigen (LNA-1/ORF73) were present in sera from all 26 KS cases and the ORF26 HHV-8 gene fragment was amplified from the tumour DNA of all patients. KS lesions that met the classical criteria for KS (spindle cells, positive endothelial markers) and were immunostained with the anti-LNA-1 marker, demonstrated typical granular intranuclear labelling of the spindle cells in all but one case of KS, in which very few spindle cells were present. Furthermore, whereas LNA-1 staining was largely confined to spindle cells in the late nodular stage, it was also observed in endothelial cells forming the walls of slit-like vessels in early patch-stage lesions. This study confirms that Morocco is an endemic area for classical KS, which remains the most frequent type of KS in this country, and emphasises the value of the LNA-1 marker for the early diagnosis of KS lesions.


Msx1 cooperates with Runx1 for inhibiting myoblast differentiation.
Myogenesis is an important and complicated biological process, especially during the process of embryonic development. The homeoprotein Msx1 is a crucial transcriptional repressor of myogenesis and maintains myogenic precursor cells in an undifferentiated, proliferative state. However, the molecular mechanism through which Msx1 coordinates myogenesis remains to be elucidated. Here, we determine the interacting partner proteins of Msx1 in myoblast cells by a proteomic screening method. Msx1 is found to interact with 55 proteins, among which our data demonstrate that the cooperation of Runt-related transcription factor 1 (Runx1) with Msx1 is required for myoblast cell differentiation. Our findings provide important insights into the mechanistic roles of Msx1 in myoblast cell differentiation, and lays foundation for the myogenic differentiation process.


Different granule cell populations innervate superficial and deep regions of the external plexiform layer in rat olfactory bulb.
Studies on the morphological organization of the main olfactory bulb have indicated that there are subpopulations of granule cells with different dendritic patterns in the external plexiform layer (EPL). Small, extracellular injections of horseradish peroxidase (HRP) were made iontophoretically into superficial and deep parts of the EPL and the granule cell layer (GCL) in adult rats. Superficial EPL injections principally labeled superficial granule cell somata, whereas deep EPL injections labeled both superficial and deep granule cell somata. Injections in the superficial GCL labeled granule cell dendritic processes extending across the entire EPL. However, deep GCL injections labeled few granule cell dendrites in the superficial EPL, but labeled many such processes in the deep EPL. These results were the same in material processed with the Hanker-Yates procedure, where the morphology of individual neurons could be studied, and in the more sensitive tetramethyl benzidine procedure. Serial reconstructions of individual granule cells were made from both HRP and Golgi-Kopsch material. The distal dendrites of deep granule cells reached only as far as the deep EPL, where they branched extensively and had many dendritic spines. The dendrites of superficial granule cells, however, reached the most superficial part of the EPL where they ramified most extensively. The superficial granule cells typically had a higher spine density in the superficial part of the EPL than in the deep part. On the basis of these results, we conclude that the superficial granule cells predominantly innervate the superficial EPL and that the deep granule cells exclusively influences on the bulbar output neurons, the mitral and tufted cells, through reciprocal dendrodendritic synapses. Since the secondary dendrites of the tufted cells ramify in the superficial EPL and the dentrites of most mitral cells ramify in deep EPL, the superficial and deep granule cells may preferentially modulate the responses of tufted and mitral cells, respectively.


Evaluation of Automated Volumetric Cartilage Quantification for Hip Preservation Surgery.
Automating the process of femoroacetabular cartilage identification from magnetic resonance imaging (MRI) images has important implications to guiding clinical care by providing a temporal metric that allows for optimizing the timing for joint preservation surgery. In this paper, we evaluate a new automated cartilage segmentation method using a time trial, segmented volume comparison, overlap metrics, and Euclidean distance mapping. We report interrater overlap metrics using the true fast imaging with steady-state precession MRI sequence of 0.874, 0.546, and 0.704 for the total overlap, union overlap, and mean overlap, respectively. This method was 3.28× faster than manual segmentation. This technique provides clinicians with volumetric cartilage information that is useful for optimizing the timing for joint preservation procedures.


Longitudinal study comparing the dynamics of Clostridium difficile in conventional and antimicrobial free pigs at farm and slaughter.
Clostridium difficile is the leading cause of nosocomial diarrhea in humans and a major cause of enteritis in neonatal piglets, foals and calves. The aim of this longitudinal study was to determine and compare the prevalence, antimicrobial susceptibility, and toxinotype profiles of C. difficile isolated from pigs and their environment in the indoor conventional and outdoor antimicrobial free (ABF) production systems. Ten conventional and eight ABF cohorts of 35 pigs each and their environment were sampled at different stages of production at farm and slaughter. C. difficile prevalence in pigs was highest at the farrowing stage in both conventional (34%, 120/350) and ABF (23%, 56/244) systems, and decreased with age. This reduction in C. difficile prevalence in pigs at later stages of production mirrored the decreased prevalence in the farm environment. At slaughter, C. difficile was isolated at a low frequency from the carcasses and processing environment in both production systems. All but three isolates were resistant to ciprofloxacin (99%, 505/508), while 1.0% (5/508) and 6.0% (23/508) of isolates exhibited resistance to tetracycline and erythromycin, respectively. Toxinotype V (tcdA(+)tcdB(+)) was the predominant strain identified in both systems (conventional: 94%, 376/401; ABF: 82%, 88/107), while the rest were toxinotype XIII (tcdA(+)tcdB(+)). To conclude, we isolated antimicrobial resistant C. difficile regardless of antimicrobial use on the farm. Based on the phenotypic and genotypic similarity of C. difficile isolated in this study, we conclude that the unique production practices employed in conventional and ABF production systems have no impact on the pathogen population.


Antibody neutralization of retargeted measles viruses.
The measles virus (MV) vaccine lineage is a promising oncolytic but prior exposure to the measles vaccine or wild-type MV strains limits treatment utility due to the presence of anti-measles antibodies. MV entry can be redirected by displaying a polypeptide ligand on the Hemagglutinin (H) C-terminus. We hypothesized that retargeted MV would escape neutralization by monoclonal antibodies (mAbs) recognizing the H receptor-binding surface and be less susceptible to neutralization by human antisera. Using chimeric H proteins, with and without mutations that ablate MV receptor binding, we show that retargeted MVs escape mAbs that target the H receptor-binding surface by virtue of mutations that ablate infection via SLAM and CD46. However, C-terminally displayed domains do not mediate virus entry in the presence of human antibodies that bind to the underlying H domain. In conclusion, utility of retargeted oncolytic measles viruses does not extend to evasion of human serum neutralization.


The structure, function and expression analysis of the nodulin 26-like intrinsic protein subfamily of plant aquaporins in tomato.
The nodulin 26-like intrinsic protein (NIP) family belonging to a group of aquaporin proteins is unique to plants. NIPs have a wide of transport activities and are involved in developmental processes and stress tolerance. The well reported Lsi1 and Lsi6 belonging to NIP III were characterized as Si transporters. However, except Lsi1 and Lsi6, most NIPs remain unknown. Here, we identified 43 putative aquaporins in tomato. We found there are 12 NIPs, including 8 NIP I proteins, 3 NIP II proteins, and 1 NIP III protein among the 43 aquaporins. Also, there are two Si efflux transporters SlLsi2-1 and SlLsi2-2 identified by using Lsi2 proteins from other species. By analysing the phylogenetic relationships, conserved residues and expression patterns, we propose that three NIP I members (SlNIP-2, SlNIP-3 and SlNIP-11) may transport water, ammonia, urea, and boric acid, and contribute to pollen development. Three NIP II proteins (SlNIP-7, SlNIP-9 and SlNIP-12) may be boric acid facilitators, and affect plant growth and anther development. Overall, the study provides valuable candidates of Si transporters and other NIP proteins to further explore their roles in uptake and transport for silicon, boron, and other substrates in tomato.


The subcoronal meatus.
There has been much recent enthusiasm for repair of subcoronal hypospadias, which is due to the positive experience with meatal advancement, glanuloplasty and its excellent cosmetic results, extremely low complication rate and brief hospitalization requirement. Although we have found the meatal advancement, glanuloplasty to be an extremely reliable procedure in the majority of boys with subcoronal hypospadias and no fibrous chordee, we have encountered 3 variants, patulous, fibrotic and hypoplastic, of the subcoronal meatus, which are not amenable to meatal advancement, glanuloplasty. We describe a logical approach to the assessment of the subcoronal meatus with use of a vascularized meatal-based flap for hypospadias repair when meatal advancement, glanuloplasty is not applicable.


Hierarchical nanocomposites with an N-doped carbon shell and bimetal core: Novel enzyme nanocarriers for electrochemical pesticide detection.
Core-shell structured nanocomposites (named PtPd@NCS) with N-doped carbon shell and bimetal core (Pt and Pd) were fabricated through a facile strategy for the first time. The PtPd@NCS nanocomposites were obtained through reduction of K2PtCl4, H2PtCl6 and Na2PdCl4 species, self-polymerization of dopamine (DA) and co-assembly of Pluronic F127 using a one-pot approach. DA serves as a reductant, as well as a carbon and nitrogen source. The core-shell structure of the PtPd@NCS nanocomposites was characterized and the result indicated that Pt-Pd nanoparticle core with a diameter of approximately 15 nm was encased in the N-doped carbon shells with a thickness of approximately 35 nm. The PtPd@NCS nanocomposites were used as an electrode material to prepare acetylcholinesterase (AChE) biosensors for detecting organophosphate pesticides. The obtained AChE biosensor exhibited a linear range of 1 × 10-14 to 1 × 10-10 M and 1 × 10-9 to 1 × 10-5 M within the detection limit of 7.9 × 10-15 M for malathion, 1 × 10-13 to 1 × 10-6 within the detection limit of 7.1 × 10-14 M for chlopyrifos, and 1 × 10-14 to 1 × 10-11 M and 1 × 10-10 to 1 × 10-5 M within the detection limit of 8.6 × 10-15 M for parathion methyl. The proposed biosensor also exhibited high selectivity, reproducibility and stability. The AChE biosensor was also applied in real samples for detecting organophosphate pesticides and exhibited acceptable recovery. This work demonstrated that the PtPd@NCS had great potential in constructing biosensors to detect organophosphate pesticides and other analytes.


Stimulation of 86Rb+ and 32Pi movements in 3T3 cells by prostaglandins and phorbol esters.
The potent tumor promoter tetradecanoyl phorbol acetate (TPA) induces early changes in ion movements analogous to those induced by prostaglandins E1 and F 2alpha. Among the earliest changes induced by TPA is a significant increase in 32Pi incorporation within 15 minutes incubation of TPA (10(-8)-10(-6) M) with post-confluent Swiss 3T3 mouse embryonic fibroblasts. Similarly, the active phorbol ester homolog 4-beta-OH phorbol didecanoate but not the inactive stereoisomeric 4-alpha-OH phorbol didecanoate stimulated 32Pi incorporation. Also, TPA at the above concentrations stimulated 86Rb+ influx shortly after administration. Both fluxes were ouabain-sensitive in accord with the idea that an early effect of TPA is to alter (Na+ + K+)-ATPase activity. Further, prostaglandin E1 (10(-7)-10(-6) M) and prostaglandin F 2alpha (3 X 10(-9)-10(-7) M) caused a similar stimulation of 86Rb+ and 32Pi uptake. The finding that water-soluble prostaglandin F 2alpha also exhibited stimulatory effects indicated that those hormone-induced responses are not mediated by solvent interactions. The similar responses of phorbol esters and prostaglandin derivatives suggests that phorbol esters and prostaglandin derivatives may act at common membrane sites. The finding that stimulatory effects were observed at discrete times in the logarithmic phase of growth suggests that the activation of membrane receptors may be cell-cycle dependent.


Colchicine as an investigative tool in neurobiology.
Neurotoxicants are being used with increasing frequency in neurobiology as investigative tools to study the structure and function of the nervous system. Colchicine administered directly into the hippocampus produces preferential destruction of dentate gyrus granule cells and mossy fibers and affects conditioned behavior. Studies from our laboratory have demonstrated the dose- and time-dependent loss of dentate gyrus granule cells and stimulation of motor activity following intradentate administration of colchicine. Preferential degeneration of pyramidal cells and other morphological changes observable at the light microscopic level were not seen in colchicine-treated rats. Other studies showed that intradentate colchicine produces specific damage to granule cells in both the dorsal and ventral hippocampus and that this damage is associated with depletion of dynorphin, a neuropeptide preferentially localized in the granule cells and mossy fibers of the hippocampus. Finally, it was noted that there are compensatory changes in the nervous system following treatment with colchicine and that the behavioral effects of colchicine can be modified by factors such as handling. One potentially important compensatory change occurring after intradentate colchicine is an alteration in cholinergic function. Pharmacological studies suggest that colchicine-treated rats may be less sensitive to the behavioral effects of scopolamine. These experiments support the conclusion that given at the appropriate dose into the hippocampus, colchicine may be a useful investigative tool to study the function of the dentate gyrus granule cells and mossy fibers, as well as the compensatory changes in the nervous system that follow chemical-induced neurodegeneration.


Robust motion correction for myocardial T1 and extracellular volume mapping by principle component analysis-based groupwise image registration.
Myocardial tissue characterization by MR T1 and extracellular volume (ECV) mapping has demonstrated clinical value. The modified Look-Locker inversion recovery (MOLLI) sequence is a standard mapping technique, but its quality can be negatively affected by motion. To develop a robust motion correction method for T1 and ECV mapping. Retrospective analysis of clinical data. Fifty patients who were referred to cardiac MR exam for T1 mapping. 3.0T cardiac MRI with precontrast and postcontrast MOLLI acquisition of the left ventricle (LV). A groupwise registration method based on principle component analysis (PCA) was developed to register all MOLLI frames simultaneously. The resulting T1 and ECV maps were compared to those from the original and motion-corrected MOLLI with pairwise registration, in terms of standard deviation (SD) error. Paired variables were compared using the Wilcoxon signed-rank test. The groupwise registration method demonstrated improved registration performance compared to pairwise registration, with the T1 SD error reduced from 31 ± 20 msec to 26 ± 15 msec (P < 0.05), and ECV SD error reduced from 4.1 ± 3.6% to 2.8 ± 2.0% (P < 0.05). In LV segmental analysis, the performance was particularly improved in lateral segments, which are most affected by motion. The running time of groupwise registration was significantly shorter than that of the pairwise registration, 17.5 ± 3.0 seconds compared to 43.5 ± 2.2 seconds (P < 0.05). We developed an automatic, robust motion correction method for myocardial T1 and ECV mapping based on a new groupwise registration scheme. The method led to lower mapping error compared to the conventional pairwise registration method in reduced execution time. 3 Technical Efficacy: Stage 1 J. Magn. Reson. Imaging 2018;47:1397-1405.


External validation of Cardiac disease, Hypertension, and Logarithmic Left anterior descending coronary artery radiation dose (CHyLL) for predicting major adverse cardiac events after lung cancer radiotherapy.
Major adverse cardiac events(MACE) are prevalent in patients with locally advanced-non-small cell lung cancer(LA-NSCLC) following radiotherapy(RT). The CHyLL model, incorporating coronary heart disease(CHD),Hypertension(HTN),Logarithmic LADV15 was developed and internally-validated to predict MACE among LA-NSCLC patients. We sought to externally validate CHyLL to predict MACE in an independent LA-NSCLC cohort. Patients with LA-NSCLC treated with RT were included. CHyLL score was calculated:5.51CHD + 1.28HTN + 1.48ln(LADV15 + 1)-1.36CHD*ln(LADV15 + 1). CHyLL performance in predicting MACE was assessed and compared to mean heart dose(MHD) using Cox-proportional hazard(PH) analyses and Harrel's concordance(C)-indices. MACE and overall survival(OS) among low-vs high-risk groups(CHyLL < 5 vs ≥ 5) were compared. In the external validation cohort(N = 102), the median age was 71 years and 55% were females. Most(n = 74,73%), had clinical Stage III disease and 35(34%) underwent surgery. CHyLL demonstrated good MACE prediction with C-index of 0.73(95% Confidence Interval(CI):0.58-0.89), while MHD did not (C-index = 0.46 (95% CI:0.30-0.62)). Per CHyLL, 32(31%) and 70(69%) patients were considered low-and high-risk for MACE, respectively. CHyLL consistently identified lower MACE rates in the low-vs high-risk group(log-rank p = 0.108):0 vs 8%(12 months),5 vs 16%(24 months),5 vs 16%(36 months),and 5 vs 19%(48 months) post-RT. In the pooled internal and external validation cohort(N = 303), MACE rates in low-vs high-risk groups were statistically significantly different(log-rank p = 0.01):1 vs 6%(12 months),3 vs 12%(24 months),6 vs 19%(36 months),and 6 vs 21%(48 months). CHyLL was externally validated and superior to MHD in predicting MACE. CHyLL has the potential to identify high-risk patients who may benefit from cardio-oncology optimization and to estimate personalized LADV15 constraints based on cardiac risk factors and acceptable MACE thresholds.


Analysis of salt-inducible genes in barley roots by differential display.
To obtain insight into the comprehensive molecular characteristics related to the mechanisms of salt tolerance, we performed a large-scale screening of salt-inducible genes in barley roots by differential display. A comparative analysis of gene expression between control and salt-stressed conditions led to the detection of 218 cDNA clones induced by salt. Sequence analysis and database searching revealed that 133 cDNA clones have homology to known proteins. Twenty-four salt-inducible clones were identified as genes for signal transduction (e.g., phosphatidylinositol-4-phosphate-5-kinase, mitogen-activated protein kinase, transcription factor, receptor protein kinase, and protein phosphatase 2A). We also detected clones encoding glutathione reductase, thioredoxin-like protein, trehalose-6-phosphate synthetase, and heat shock proteins in the category of typical stress tolerance. Furthermore, we have obtained genes encoding membrane transporters, members of the P450 family, enzymes involved in RNA metabolism or function, and enzymes of sugar or amino acid metabolism. It must be noted that most genes were expressed strongly in roots, but only rarely or weakly in leaves. In addition, some clones were newly found as salt-inducible genes encoding SCARECROW, splicing factor and apoptosis protein. In this research, it was shown that differential display is a powerful tool for a large-scale cloning of cDNAs induced by salt and these results are very useful for understanding the mechanisms of plant salt tolerance.


Predictive significance for sudden death of microvolt-level T wave alternans in New York Heart Association class II congestive heart failure patients: a prospective study.
Sudden cardiac death (SDC) is responsible for approximately 60-70% of deaths in New York Heart Association (NYHA) class II congestive heart failure (CHF) patients. Recently, microvolt-level T wave alternans has been proposed as a new noninvasive tool to identify CHF patients at risk for SCD and ventricular tachycardia/fibrillation (VT/VF). To determine the prognostic value of MTWA in NYHA class II patients. Among 181 consecutive CHF patients with ischemic and nonischemic cardiomyopathy, 73 patients in NYHA class II with left ventricular ejection fraction <45% were selected and prospectively investigated. MTWA was determined during bicycle exercise testing. The study end point was defined as SCD, documented sustained VT/VF and appropriate implantable cardioverter defibrillator (ICD) shock. MTWA was positive in 30 (41%) patients, negative in 26(36%) patients and indeterminate in 17 (23%) patients. During an average follow-up of 17.1+/-7.4 months, seven patients had an arrhythmic event in the MTWA positive group, whereas one and no events occurred in the indeterminate and negative group, respectively. From Kaplan-Meier univariate analysis and multivariate Cox analysis, MTWA was a significant arrhythmic risk stratifier (p=0.01 and p=0.03, respectively). Sensitivity, specificity, negative and positive predictive values of MTWA were 100%, 53%, 100% and 24%, respectively. Our data suggest that MTWA is a promising predictor of arrhythmic events in NYHA class II CHF patients.


A method to assess the default EEG macrostate and its reactivity to stimulation.
The default mode network (DMN) is deactivated by stimulation. We aimed to assess the DMN reactivity impairment by routine EEG recordings in stroke patients with impaired consciousness. Binocular light flashes were delivered at 1 Hz in 1-minute epochs, following a 1-minute baseline (PRE). The EEG was decomposed in a series of binary oscillatory macrostates by topographic spectral clustering. The most deactivated macrostate was labeled the default EEG macrostate (DEM). Its reactivity (DER) was quantified as the decrease in DEM occurrence probability during stimulation. A normalized DER index (DERI) was calculated as DER/PRE. The measures were compared between 14 healthy controls and 32 comatose patients under EEG monitoring following an acute stroke. The DEM was mapped to the posterior DMN hubs. In the patients, these DEM source dipoles were 3-4 times less frequent and were associated with an increased theta activity. Even in a reduced 6-channel montage, a DER below 6.26% corresponding to a DERI below 0.25 could discriminate the patients with sensitivity and specificity well above 80%. The method detected the DMN impairment in post-stroke coma patients. The DEM and its reactivity to stimulation could be useful to monitor the DMN function at bedside.


Characteristics and outcomes of heart failure-related hospitalization in adults with congenital heart disease.
Heart failure (HF) is the main cause of death in adult congenital heart disease (ACHD). We aimed to characterize HF-related hospitalization of patients with ACHD, and to determine HF risk factors and prognosis in this population. We prospectively included 471 patients with ACHD admitted to our unit over 24 months. Clinical and biological data and HF management were recorded. Major cardiovascular events were recorded for ACHD with HF. HF was the main reason for hospitalization in 13% of cases (76/583 hospitalizations). Patients with HF were significantly older (median age 44±14 years vs. 37±15 years; P<0.01), with more complex congenital heart disease (P=0.04). In the multivariable analysis, pulmonary arterial hypertension (odds ratio [OR] 6.2, 95% confidence interval [CI] 3.5-10.7), history of HF (OR 9.8, 95% CI 5.7-16.8) and history of atrial arrhythmia (OR 3.6, 95% CI 2.2-5.9) were significant risk factors for HF-related admissions (P<0.001). The mean hospital stay of patients with HF was longer (12.2 vs. 6.9 days; P<0.01), and 25% of patients required intensive care. Overall, 11/55 (20%) patients with HF died, 10/55 (18%) were readmitted for HF, and 6/55 (11%) had heart transplantation during the median follow-up of 18 months (95% CI 14-20 months). The risk of cardiovascular events was 19-fold higher after HF-related hospitalization. HF is emerging as a leading cause of morbidity and mortality in the ACHD population. Earlier diagnosis and more active management are required to improve outcomes of HF in ACHD.


Advances of circRNA-miRNA-mRNA regulatory network in cerebral ischemia/reperfusion injury.
Ischemic stroke (IS) is the most common type of stroke, and its pathophysiological process is more complex. In recent years, the key regulatory roles of non-coding RNA (miRNA, circRNA) and mRNA in the development of IS have attracted more attention. In the process of cerebral ischemia/reperfusion injury, circRNA can regulate nerves, blood vessels and immune system through miRNA/mRNA axis, so as to affect the neurovascular unit of IS. The combination of these noncoding RNAs and mRNAs can be used as non-invasive biomarkers and therapeutic tools for IS diagnosis, prognosis and brain injury. Therefore, it is very important to study the potential molecular mechanism, activation pathway and treatment methods of circRNA/miRNA/mRNA network in IS. This review will focus on the latest progress of circRNA/miRNA/mRNA regulatory network, we have also included some circRNAs, which does not mediate through a miRNA, so we also include circRNA -mRNA network. And explore the application prospect of these RNAs as potential therapeutic targets in the prevention and treatment of IS.


Effects of sulfur-based hemostatic agents and gingival retraction cords handled with latex gloves on the polymerization of polyvinyl siloxane impression materials.
This study investigated the possible interactions between three addition silicone materials (Express®, Aquasil Ultra® and Adsil®), three hemostatic agents (ferric sulfate, StatGel FS®; aluminum sulfate, GelCord®; and aluminum chloride, Hemostop®) and gingival retraction cords previously handled with latex gloves to determine whether direct contact with medicaments or indirect contamination by latex in conditions similar to those found in clinical practice inhibit or affect the setting of the impression materials. A portable device for the simultaneous test of several specimens was specifically developed for this study. Polymerization inhibition was analyzed by examination of the impressions and the molded surface. Ten trials were performed for each addition silicone material used in the study, at a total of 240 study samples. All the samples tested (N=240) were nonreactive regardless of the type of combination used. Aluminum sulfate, ferric sulfate and aluminum chloride hemostatic solutions did not show any inhibitory potential on the addition silicone samples under study, and there were no changes in polymerization as a result of contact between addition silicone and retraction cords handled with latex gloves.


Invited review: Antimicrobial resistance in bovine mastitis pathogens: A review of genetic determinants and prevalence of resistance in European countries.
Antimicrobial resistance is an urgent and growing problem worldwide, both for human and animal health. In the animal health sector actions have been taken as concerns grow regarding the development and spread of antimicrobial resistance. Mastitis is the most common infection in dairy cattle. We aimed to summarize the genetic determinants found in staphylococci, streptococci, and Enterobacteriaceae isolated from mastitic milk samples and provide a comparison of percentage resistance to a variety of antimicrobials in European countries.


Role of milk-derived IgG in passive maternal protection of neonatal ferrets against influenza.
Neonatal ferrets are protected against infection with influenza virus by colostral and milk-derived anti-influenza virus IgG after suckling on an immune mother. The levels of IgG elicited and then transmitted to neonates were similar when mothers were immunized by either live infection or killed vaccines. Maternal anti-influenza virus IgA and IgM appears not to cross the neonatal gut epithelium although both are present in maternal serum and milk.


Biofilm on ventriculo-peritoneal shunt tubing as a cause of treatment failure in coccidioidal meningitis.
We describe a case of recurrent coccidioidal meningitis in which a fungal biofilm on the tip of ventriculo-peritoneal shunt tubing was likely responsible for a 4-year persistence of Coccidioides immitis, despite the patient's taking an adequate dosage of fluconazole. Fungal biofilms should be considered as a cause for treatment failure and fungal persistence, especially when artificial prostheses or indwelling catheters are present.


Arthroscopic double-button Latarjet osteolysis and remodeling at 1-year follow-up.
The aim of this study was to evaluate the osteolysis rate, graft remodeling, and risk factors for osteolysis at the 1-year threshold after an arthroscopic Latarjet procedure with double-button fixation. In this multicenter, retrospective study, postoperative computed tomography scans obtained after an arthroscopic Latarjet procedure with double-button fixation to treat anterior shoulder instability were analyzed at 15 days and at 3, 6, and 12 months. Graft volume, dimensions, and morphologic remodeling were analyzed. Twenty-seven patients were included (mean age, 26 years). At 1 year, osteolysis occurred in 19 of 27 patients (70%). The volume initially decreased until 6 months' follow-up (-35%; range, -75% to +26%) and then increased until our last follow-up. At 1 year, the graft volume decreased by 17% (range, -61% to +56%) compared with the immediate postoperative volume. In multivariate analysis, the rate of osteolysis was inversely associated with an unhealed graft at 3 months (P = .02; β coefficient = -44.50 [95% confidence interval, -81 to -8]). The maximal height of the graft significantly grew 0.2 cm (range, -55 to +124 mm) (P = .015). In the sagittal plane, osteolysis occurred in the superior part in 100% of patients (27 of 27) whereas bone formation occurred in the inferior part. In the axial plane, osteolysis occurred in the anterior part whereas bone formation occurred in the posterior part. In the articular part, the observed remodeling was aimed to obtain a new anatomic and congruent glenoid. At 1 year after an arthroscopic Latarjet procedure with double-button fixation, osteolysis occurred in 70% of patients. The rate of osteolysis was 17% of the initial volume. Osteolysis occurred mainly during the first 6 months, in the anterior and superior parts. Remodeling led to a circular anatomic glenoid. This osteolysis did not cause any recurrence of instability or require revision surgery.


Prognostic and predictive impacts of tumor-infiltrating lymphocytes differ between Triple-negative and HER2-positive breast cancers treated with standard systemic therapies.
Tumor-infiltrating lymphocytes (TILs) have potential value for stratifying the treatment of breast cancer (BC), though their precise use remains unclear. We aimed to investigate the utility of TILs using an alternative approach in different settings. We reviewed patients with triple-negative (TN) or human epithelial growth factor receptor 2 (HER2)-positive invasive ductal carcinomas from a single institutional cohort and classified archived hematoxylin-eosin-stained samples in terms of TIL score as low (<10 %), intermediate, and high (>50 %). The prognostic and predictive values of TILs were analyzed retrospectively in both adjuvant and neo-adjuvant settings. In the adjuvant setting, the presence of TILs at primary surgery was a significant favorable prognostic factor among 154 TNBCs [relapse-free survival (RFS): p = 0.015], but not among 183 HER2+ BCs (RFS: p = 0.097). The TNBC low-TIL group tended to relapse earlier. In the neo-adjuvant setting, detection of TILs on biopsy before primary systemic therapy was associated with the ratio of patients achieving pathological complete response among 48 TNBCs (p = 0.024), but not among 58 HER2+ BCs (p = 0.30). The presence of TILs in surgical specimens after systemic therapy had prognostic value in HER2+ BC (RFS: p = 0.007). The impact of TILs differs between patients with TN and HER2+ BC treated with standard therapies. Our three-grade scale for TILs may contribute to our understanding of the importance of the tumor microenvironment in routine practice. TILs after primary systemic therapy may be useful for the further stratification of treatment of HER2+ BC.


Electromagnetic irradiation may be a new approach to therapy for peri-implantitis.
Peri-implantitis can lead to bone destruction around a dental implant through inflammation and immune reactions caused by bacteria adhering to the surface of the implant abutment. Electromagnetic irradiation can inhibit bacterial growth, increase bone formation, decrease bone resorption and reduce the inflammatory response. Our hypothesis is that electromagnetic irradiation may be a new treatment approach for peri-implantitis and may simultaneously maintain bone mass around the dental implant. The results would be more significant when combined with other agents, because the effect of some antibiotics and anti-inflammatory drugs is strengthened by electromagnetic irradiation. This non-invasive therapy is expected to be conducted in a convenient manner, and even by patients at home, thereby facilitating the prevention and treatment of peri-implantitis.


The utility of Plasmodium berghei as a rodent model for anti-merozoite malaria vaccine assessment.
Rodent malaria species Plasmodium yoelii and P. chabaudi have been widely used to validate vaccine approaches targeting blood-stage merozoite antigens. However, increasing data suggest the P. berghei rodent malaria may be able to circumvent vaccine-induced anti-merozoite responses. Here we confirm a failure to protect against P. berghei, despite successful antibody induction against leading merozoite antigens using protein-in-adjuvant or viral vectored vaccine delivery. No subunit vaccine approach showed efficacy in mice following immunization and challenge with the wild-type P. berghei strains ANKA or NK65, or against a chimeric parasite line encoding a merozoite antigen from P. falciparum. Protection was not improved in knockout mice lacking the inhibitory Fc receptor CD32b, nor against a Δsmac P. berghei parasite line with a non-sequestering phenotype. An improved understanding of the mechanisms responsible for protection, or failure of protection, against P. berghei merozoites could guide the development of an efficacious vaccine against P. falciparum.