Killing of brain tumor cells by hypoxia-responsive element mediated expression of BAX.
The presence of radioresistant hypoxic cells in human brain tumors limits the overall effectiveness of conventional fractionated radiation therapy. Tumor-specific therapies that target hypoxic cells are clearly needed. We have investigated the expression of suicide genes under hypoxia by a hypoxia-responsive element (HRE), which can be activated through hypoxia-inducible factor-1 (HIF-1). We transfected plasmids containing multiple copies of HRE into U-87 MG and U-251 MG-NCI human brain tumor cells and tested their ability to induce LacZ gene expression under anoxia. Gene expression under anoxia versus oxia was increased about 12-fold for U-87 MG cells and about fourfold for U-251 MG-NCI cells. At intermediate hypoxic conditions, increased LacZ gene expression in U-87 MG cells was induced by the plasmid that contained three HREs, but not by the plasmid with two HREs. Lastly, when we placed a suicide gene BAX under the control of HREs, cells transfected with the BAX plasmids were preferentially killed through apoptosis under anoxia. Our studies demonstrate that HRE-regulated gene expression is active in brain tumor cells, and that the amount of increased gene expression obtained is dependent on the cell line, the HRE copy number, and the degree of hypoxia.


Comparison of safety outcomes of anticancer drugs in Japanese and non-Japanese patients in multi-regional clinical trials: meta-analysis of safety profiles.
No report has assessed the differences in adverse event (AE) profiles of anticancer drugs for all types of cancers in clinical trials involving Japanese patients. This study aimed to compare the safety outcomes of anticancer drugs in Japanese and non-Japanese patients in multi-regional clinical trials (MRCTs), regardless of the type of cancer or drug. All new approvals of oncology drugs in Japan from January 2009 to December 2018 were searched through the Pharmaceuticals and Medical Devices Agency website. The odds ratio (OR) for comparing the incidence of AEs between Japanese and non-Japanese patients was estimated using the Mantel-Haenszel method with a random effect model. Sixty-six multi-regional phase 3 trials were identified involving 43,712 patients. Severe AE, AE leading to dose reduction, and AE leading to dose interruption were significantly more frequent in Japanese patients than in non-Japanese (odds ratios [ORs] were 1.32 (95% confidence interval (CI): 1.13-1.53), 1.97 (95% CI: 1.66-2.31), and 1.63 (95% CI: 1.43-1.86), respectively). Serious AEs (SAEs) and AEs leading to death were significantly less frequent in Japanese patients (OR: 0.70 (95% CI: 0.62-0.77) vs. 0.56 (95% CI: 0.44-0.67), respectively). There were no differences in AEs leading to study withdrawal. The incidence of most AEs was significantly higher in Japanese patients. In Japanese patients, the incidence of SAEs and AEs leading to death was low, but severe AEs, AEs leading to dose reductions, AEs leading to dose interruption, and individual adverse events were high.


Biliary atresia: early determination of prognosis.
Analyses of bile bilirubin during the first month after Kasai operations, and of the liver biopsies obtained at the time of initial surgery, were done in 67 patients with biliary atresia. Bilirubin excretion (milligrams per day) was determined as the product of the bile volume and its concentration. Operative liver biopsies were evaluated for fibrosis, bile duct proliferation, bile stasis, giant cell transformation, and parenchymal degeneration; the severity of each abnormality was graded on a scale of 0 to 4. A forward stepwise regression procedure using the Cox proportional hazards model identified the relationship between survival and covariants. Thirty-nine of 67 patients died. Of these, 38 excreted less than 6 mg of bilirubin per day during the first postoperative month. The other patient died of a coexisting anomaly. Nine other patients who excreted less than 6 mg of bilirubin per day are alive but are either jaundiced or awaiting transplantation. Nineteen patients who excreted greater than or equal to 6 mg of bilirubin per day are alive (mean follow-up, 61 months) with normal or near-normal liver function. The severity of liver fibrosis, bile duct proliferation, and bile stasis did not correlate with survival, whereas giant cell transformation and parenchymal degeneration were highly significant (P less than or equal to .000 and P less than or equal to .0003, respectively). Twenty-six infants with grade 1-4 giant cell transformation or grade 2-4 parenchymal degeneration had a mean survival of 11 months.(ABSTRACT TRUNCATED AT 250 WORDS)


Age structure and feeding of the neotropical grasshopper Cornops aquaticum (Bruner) (Orthoptera: Acrididae) on water hyacinth.
We aimed to evaluate the variation in the age structure of Cornops aquaticum (Bruner) population and its relation to the host plant biomass and the feeding of the different age classes of this grasshopper on the water hyacinth Eichhornia crassipes along 2 years, in a Paraná River floodplain lake (Chaco, Argentina). Individuals of C. aquaticum were captured with a 70-cm diameter sweep and separated in nymphs A (instars I and II), nymphs B (instars III to VI), adult females, and adult males; host plant biomass was sampled using a ring with a 0.30-m(2) diameter. Relative daily feeding of C. aquaticum population was calculated by multiplying the number of individuals captured per minute by the daily consumption by individual obtained in each age classes. We found that the age structure and the relative daily feeding of C. aquaticum varied between seasons and years. The highest values of grasshopper abundance, leaf biomass, and relative daily feeding of C. aquaticum population were observed in summer 2006. Plant biomass was directly correlated with nymph abundance and not correlated with adult abundance. Plant biomass available as refuge (leaves), food (laminas), and oviposition site (petioles) to C. aquaticum represented up to 62% of the total plant biomass. The results obtained in C. aquaticum show the importance of considering total plant biomass and plant biomass available for herbivores separately. Our study highlights the need to find an adequate method to estimate the density of C. aquaticum and other semiaquatic grasshoppers in the Paraná River floodplain involving different seasons, years, and water phases (rising and falling).


Extracting plants core genes responding to abiotic stresses by penalized matrix decomposition.
Sparse methods have a significant advantage to reduce the complexity of genes expression data and to make them more comprehensible and interpretable. In this paper, based on penalized matrix decomposition (PMD), a novel approach is proposed to extract plants core genes, i.e., the characteristic gene set, responding to abiotic stresses. Core genes can capture the changes of the samples. In other words, the features of samples can be caught by the core genes. The experimental results show that the proposed PMD-based method is efficient to extract the core genes closely related to the abiotic stresses.


The Trainee's Role in Patient Safety: Training Residents and Medical Students in Surgical Patient Safety.
"The focus on patient safety offers a new framework not only for delivering health care but also for training physicians. Medical school and surgical graduate medical education must transition to a more holistic approach by teaching technical and nontechnical skills. Formalized safety curricula can be developed by adopting recommended guidelines and content from national and international organizations, existing validated practices of training programs, frequent simulation exercises, and objective evaluation tools."


Immunological studies of human monoamine oxidases.
Antisera have been raised against monoamine oxidase preparations from human placenta and platelets. These antisera have been employed to characterize membrane-bound enzyme from a variety of human sources including liver, heart, and brain. The comparisons were based on a displacement radioimmunoassay system with soluble placental monoamine oxidase, previously labelled specifically with [3H]pargyline, as antigen. All forms of enzyme investigated demonstrated immunological cross reaction; however, the placental enzyme appeared to possess determinants not exhibited by the enzyme from the platelets or other tissues examined.


Ultrastructure of chemically defined neuron systems in the dorsal horn of the monkey. I. Substance P immunoreactivity.
The ultrastructural localization of substance P-like immunoreactivity (SPLI) in lamina I (marginal zone) and lamina IIO (outer substantia gelatinosa) of the dorsal horn of the macaque monkey was examined by the indirect antibody peroxidase-antiperoxidase method. SPLI was found in small unmyelinated and finely myelinated axons and a variety of terminal types. The majority of SPLI terminals contained a few to many large granular vesicles (mean diameter 90 nm) in addition to a population of small clear round vesicles. A very few terminals contained mainly small round vesicles. SPLI terminals were presynaptic in axosomatic, axodendritic and axospinous contacts forming, in all but the axosomatic junctions, asymmetrical synapses. Some axosomatic junctions were symmetrical. SPLI terminals also formed the center of glomeruli with unlabeled dendrites and dendritic spines; some of the unlabeled dendrites contained a few small scattered vesicles and large dense-core vesicles. In more complex formations 2 to 4 SPLI terminals were associated with one another and linked by desmosomal contacts. The individual terminals in the complexes or 'congregate terminals' were simple large granular vesicle containing terminals (LGV), LGV-central terminals of associated glomeruli, or terminals containing mainly small round vesicles. In the apical region of lamina I an unlabeled terminal was found occasionally in contact with an SPLI terminal, which in turn synapsed onto a dendrite. These contacts have some synaptic characteristics and the SPLI terminal was possibly postsynaptic. Most of the types of SPLI terminals resemble closely terminal types shown to be of primary afferent origin. These terminals which make direct contact with dorsal horn dendrites may be the morphological substrate for postsynaptic excitation of dorsal horn neurons by substance P. The contacts of unlabeled terminals with SPLI terminals may represent a morphological substrate by which other neurochemical substances such as enkephalin or serotonin may modulate the substance P effects on dorsal horn neuronal activity.


A bioartificial liver--state of the art.
End-stage liver disease is treated by liver transplantation, but donor organ shortages remain a serious problem. This has prompted the design of bioartificial liver devices to "bridge" patients until they either recover or receive a liver transplant. In these devices, patient plasma is circulated extracorporeally through a bioreactor that houses liver cells (hepatocytes) sandwiched between artificial plates or capillaries.


Are breast tumours innervated? Immunohistological investigations using antibodies against the neuronal marker protein gene product 9.5 (PGP 9.5) in benign and malignant breast lesions.
The aim of the present study was to assess the innervation pattern of benign and malignant breast lesions using the neuronal marker protein gene product (PGP) 9.5. An unlabelled antibody technique (using streptavidin biotin complex formation) was used on paraffin wax sections of tissues fixed in neutral buffered formalin. In 2/4 cases of chronic mastopathy, PGP 9.5 immunoreactivity was seen in relation to blood vessels and the ductal system. No immunoreactivity for PGP 9.5 was seen in the affected tissues of 9/10 cases of fibroadenomata. In 9/16 breast cancers, PGP 9.5-labelled perivascular nerve fibres were detected in connective tissue stroma supporting carcinoma tissue, though not in the immediate vicinity of such tumour tissue. Labelled nerve fibres were detected in large bundles at the periphery of tumours, possibly unrelated to the latter. Our results indicate that the newly formed blood vessels within a tumour are not innervated, though major blood vessels which supply the tumour are innervated.


Subcellular distribution of ubiquitin-protein conjugates in the hippocampus following transient ischemia.
Ubiquitin-protein conjugates in the hippocampus were analyzed by immunoblotting with a monoclonal anti-ubiquitin antibody. In the CA1 region, Triton X-100 insoluble ubiquitin-protein conjugates increased after 24 hr following 20 min of ischemia. When the total hippocampi were fractionated subcellularly, ubiquitin-protein conjugates increased in the particulate, especially in the mitochondrial fraction. The ubiquitin-protein conjugates were solubilized by SDS, or were partially solubilized by urea. The results indicate that insoluble ubiquitin-protein conjugates increase after ischemia.


Biologically active phorbol esters specifically alter affinity of epidermal growth factor membrane receptors.
TPA (12-O-tetradecanoyl-phorbol-13-acetate) reversibly inhibits the binding of (125)I-labelled epidermal growth factor (EGF) to treated mouse and human cells, but does not affect the binding of various other ligands to their membrane receptors. It alters the affinity of the receptors for EGF without changing the total number of available receptors per cell. Those phorbol esters which stimulate cell growth in culture and have tumour-promoting activity in vivo alter the EGF-receptor affinity, while the biologically inactive derivatives fail to change the affinity of EGF for its receptors.


Dosing and antipyretic efficacy of oral acetaminophen in children.
A standardized approach to dosing acetaminophen in pediatric populations was published in 1983. That review proposed specific weight-related dosing for infants and children weighing 6 through 95 lb and an age-based schedule for children aged <4 months through 11 years. Subsequent clinical studies evaluating these and alternative doses of acetaminophen supported the recommended 10-15-mg/kg dose. This article reviewed published and unpublished pediatric antipyretic data to provide a critical assessment of the 10-15-mg/kg oral dose and the current pediatric oral dosing schedules for acetaminophen. Published literature and unpublished clinical trials that evaluated the antipyretic efficacy of acetaminophen in children were reviewed. The PubMed database was searched using the term acetaminophen or paracetamol, with study criteria limited to randomized, controlled trials; oral dosing; patient age <12 years; and publication between 1982 and August 2012. All of the sponsor's unpublished antipyretic clinical studies completed between 1980 and August 2012 and involving at least 1 oral-formulation acetaminophen-only treatment arm were identified. Data from published literature containing sufficient detail to verify doses; dosing frequency; and, when necessary, estimates from figures, and from acetaminophen arms of the unpublished studies were analyzed. Thirteen unpublished trials enrolled 705 children to receive an oral dose of 10-15 mg/kg of acetaminophen. This dose resulted in a rapid onset of temperature reduction, with a maximum temperature decrement of ~3 hours following administration. Results from 40 published clinical trials in which 2332 children received oral acetaminophen for fever support these findings. The most common adverse events reported in any of the reported studies were gastrointestinal in nature and generally mild in intensity. Data support the recommended 10-15-mg/kg oral dose and demonstrate that the age and weight schedules for over-the-counter acetaminophen proposed in 1983 remain appropriate.


Occurrence of priority organic pollutants in Strymon river catchment, Greece: inland, transitional, and coastal waters.
Twenty-five sampling stations were selected in order to monitor persistent organic pollutants (polycyclic aromatic hydrocarbons (PAHs), organochlorine (OC) pesticides and total polychlorinated biphenyls (PCBs)) in surface water from Kerkini Lake, the Strymon River, its main tributaries and estuary in N. Aegean Sea during January to July, 2008, according to recent European Union (EU) guidelines. The data were divided among the high (January to April) and the low flow season (May to July). Generally, the values for organic pollutants were within the range reported worldwide for surface water. Elevated PAHs concentrations were observed compared with other places in Greece. Anthracene and benzo(a)pyrene exceeded maximum allowable concentration (MAC) of the relative EU guideline. Also, concentrations above MAC were observed for OCs, γ-HCH, and a-endosulfan. Despite the fact that it is banned since 1972, Aldrin was detected during the monitoring season (from limit of detection (LOD) to 15 ng L(-1)). Total PCB concentrations ranged from LOD to 162 ng L(-1). In addition, the load of organic pollutants was estimated in April (high flow) and June (low flow) in selected sampling stations. According to this estimation, napthalene, anthracene, and fluoranthene (PAHs), total dichlorodiphenyltrichloroethane (DDT), aldrin, and total PCBs had the highest load. Taking into account the relative EU guidelines concerning the pollutants studied, the water quality in the Strymon River catchment could be characterized as poor, which can lead to negative impacts to its biota.


A network of grassroots reserves protects tropical river fish diversity.
Intensive fisheries have reduced fish biodiversity and abundance in aquatic ecosystems worldwide1-3. 'No-take' marine reserves have become a cornerstone of marine ecosystem-based fisheries management4-6, and their benefits for adjacent fisheries are maximized when reserve design fosters synergies among nearby reserves7,8. The applicability of this marine reserve network paradigm to riverine biodiversity and inland fisheries remains largely untested. Here we show that reserves created by 23 separate communities in Thailand's Salween basin have markedly increased fish richness, density, and biomass relative to adjacent areas. Moreover, key correlates of the success of protected areas in marine ecosystems-particularly reserve size and enforcement-predict differences in ecological benefits among riverine reserves. Occupying a central position in the network confers additional gains, underscoring the importance of connectivity within dendritic river systems. The emergence of network-based benefits is remarkable given that these reserves are young (less than 25 years old) and arose without formal coordination. Freshwater ecosystems are under-represented among the world's protected areas9, and our findings suggest that networks of small, community-based reserves offer a generalizable model for protecting biodiversity and augmenting fisheries as the world's rivers face unprecedented pressures10,11.


In vivo inflammatory stimulation induces a transient change in the binding of thrombin to rat peritoneal macrophages.
The binding of 125I-labeled thrombin to rat peritoneal macrophages isolated 20 h after the ip injection of thioglycollate broth or lipopolysaccharide decreased to 20% of the value found in resident macrophages due to a decrease in the number of receptors. The binding returned to normal values within a week after the injection. The decline parallelled more or less the Vmax for the 5'-nucleotidase activity. This decrease in the binding of thrombin could not be explained by an immigration of monocytes into the peritoneal cavity, since the binding of 125I-labeled alpha 2-macroglobulin-trypsin complex increased 4.5-fold in the same cell population due to an increase in the number of receptors, and blood monocytes do not bind alpha 2-macroglobulin-trypsin complex. The increase in the binding of alpha 2-macroglobulin-protease complex parallelled an increase in the incorporation of glucosamine, although the latter did not increase to the same extent. Engulfment of plasma membrane after phagocytosis did not result in a decreased binding of thrombin, but preincubation at 37 degrees C with concanavalin A caused a minor reduction in the binding. There was a positive correlation between the binding of alpha 2-macroglobulin-trypsin complex and the fraction of polymorphonuclear leukocytes in the peritoneal exudate and a negative correlation between the binding of thrombin and the fraction of polymorphonuclear leukocytes in the exudate, when the inflammation was induced by a milder stimulus, sterile NaCl, indicating a common signal for the polymorphonuclear leukocyte chemotaxis and the macrophage differentiation.


Force Control Strategy of Five-Digit Precision Grasping With Aligned and Unaligned Configurations.
To investigate the digit force control during a five-digit precision grasp in aligned (AG) and unaligned grasping (UG) configurations. The effects of various cylindrical handles for tools on power grasp performance have been previously investigated. However, there is little information on force control strategy of precision grasp to fit various grasping configurations. Twenty healthy young adults were recruited to perform a lift-hold-lower task. The AG and UG configurations on a cylindrical simulator with force transducers were adjusted for each individual. The applied force and moment, the force variability during holding, and force correlations between thumb and each finger were measured. No differences in applied force, force correlation, repeatability, and variability were found between configurations. However, the moments applied in UG were significantly larger than those in AG. The force control during precision grasp did not change significantly across AG and UG except for the digit moment. The simulator is controlled efficiently with large moment during UG, which is thus the optimal configuration for precision grasping with a cylindrical handle. Further research should consider the effects of task type and handle design on force control, especially for individuals with hand disorders. To design the handle of specific tool, one should consider the appropriate configuration according to the task requirements of precision grasping to reduce the risk of accumulating extra loads on digits with a cylindrical handle.


Investigation on "Excimer" Formation Mechanism of Linear Oligofluorenes-Functionalized Anthracenes by Using Transient Absorption Spectroscopy.
We study the photophysical characters of two oligofluorenes-functionalized anthracenes molecules with different fluorine-vinylene (FV) units, which exhibits that "excimer" state appears in the solution after photoexcitation. The dynamic data shows that two mechanisms are responsible for the generation of "excimer". The fast one is controlled by the arene-arene interaction between molecules and the slow one is influenced by the diffusion motion of molecules. Increasing the number of FV units may suppress the DM-dependent "excimer" and enhance the yield of intrinsic fluorescence, which finally improves the fluorescence property of molecules in solution.


Supraspinal-selective TRPV1 desensitization induced by intracerebroventricular treatment with resiniferatoxin.
The transient receptor potential vanilloid type 1 (TRPV1) is a thermosensitive cation channel that triggers heat pain in the periphery. Long-term desensitization of TRPV1, which can be induced by excess amounts of agonists, has been a method for investigating the physiological relevance of TRPV1-containing neuronal circuits, and desensitization induced by various routes of administration, including systemic, intrathecal and intraganglionic, has been demonstrated in rodents. In the present study, we examined the effect of intracerebroventricular (i.c.v.) treatment with an ultrapotent TRPV1 agonist, resiniferatoxin (RTX), on nociception and the analgesic effect of acetaminophen, which is known to mediate the activation of central TRPV1. I.c.v. administration of RTX a week before the test did not affect the licking/biting response to intraplantar injection of RTX (RTX test), suggesting that such i.c.v. treatment spares the function of TRPV1 at the hindpaw. Mice that had been i.c.v.-administered RTX also exhibited normal nociceptive responses in the formalin test and the tail pressure test, but acetaminophen failed to induce analgesia in those mice in any of the tests. These results suggest that i.c.v. administration of RTX leads to brain-selective TRPV1 desensitization in mice.


Confined Synthesis of Amorphous Al2 O3 Framework Nanocomposites Based on the Oxygen-Potential Diagram as Sulfur Hosts for Catalytic Conversion.
Sulfur cathodes in Li-S batteries suffer significant volumetric expansion and lack of catalytic activity for polysulfide conversion. In this study, a confined self-reduction synthetic route is developed for preparing nanocomposites using diverse metal ions (Mn2+ , Co2+ , Ni2+ , and Zn2+ )-introduced Al-MIL-96 as precursors. The Ni2+ -introduced Al-MIL-96-derived nanocomposite contains a "hardness unit", amorphous aluminum oxide framework, to restrain the volumetric expansion, and a "softness unit", Ni nanocrystals, to improve the catalytic activity. The oxygen-potential diagram theoretically explains why Ni2+ is preferentially reduced. Postmortem microstructure characterization confirms the suppressive volume expansion. The in situ ultraviolet-visible measurements are performed to probe the catalytic activity of polysulfide conversion. This study provides a new perspective for designing nanocomposites with "hardness units" and "softness units" as sulfur or other catalyst hosts.


Occurrence of Inborn Errors of Metabolism in Newborns, Diagnosis and Prophylaxis.
Inborn errors of metabolism (IEM) are a heterogeneous group of rare genetic disorders that are generally transmitted as autosomal or X-linked recessive disorders. These defects arise due to mutations associated with specific gene(s), especially the ones associated with key metabolic enzymes. These enzymes or their product(s) are involved in various metabolic pathways, leading to the accumulation of intermediary metabolite(s), reflecting their toxic effects upon mutations. The diagnosis of these metabolic disorders is based on the biochemical analysis of the clinical manifestations produced and their molecular mechanism. Therefore, it is imperative to devise diagnostic tests with high sensitivity and specificity for early detection of IEM. Recent advances in biochemical and polymerase chain reaction-based genetic analysis along with pedigree and prenatal diagnosis can be life-saving in nature. The latest development in exome sequencing for rapid diagnosis and enzyme replacement therapy would facilitate the successful treatment of these metabolic disorders in the future. However, the longterm clinical implications of these genetic manipulations is still a matter of debate among intellectuals and requires further research.


Can the uterine cervix grow a moustache?
The exceptional founding of a sebaceous gland in the squamous epithelium of the uterine cervix are described. The possible embriologic origin are discussed.


Effects of methylxanthines on the fetus.
The role of caffeine and other methylxanthines in prematurity, intrauterine growth retardation, and the respiratory distress syndrome is currently under investigation. Their pharmacokinetics are discussed together with their newly discovered beneficial role as possible inhibitors of premature labor and accelerators of fetal lung maturation.


Characterization of volatile metabolites associated with confinement odour during the shelf-life of vacuum packed lamb meat under different storage conditions.
Confinement odour was investigated. Volatiles were extracted directly from the pack, using solid phase microextraction and analysed by gas chromatography-mass spectrometry. Sensory evaluation and microbiological analysis of the meat surface were also performed. Commercial samples of vacuum packed lamb legs (n=85), from two meat processing plants, were kept for 7weeks at -1.5°C then at different regimes of temperature (-1.5 to +4°C) until 11, 12 or 13weeks. Persistent odour was observed in 66% of samples, confinement odour in 24% and no odour in 11%. Volatiles associated with confinement odour (3-methyl-butanal, 3-hydroxy-2-butanone and sulphur dioxide) corresponded with end/sub products of glucose fermentation and catabolism of amino acids by bacteria (all bacteria naturally found in meat and do not represent a risk to health). Confinement odour could indicate a stage at which the environment for bacteria growth is becoming favourable for the production of volatiles with strong odours that are noticed by the consumer.


Lymphoproliferative and myeloproliferative disorders.
Nomenclature regarding neoplasia of the hematopoietic and lymphoid tissues in the horse is confusing. This article will clarify terminology, and discuss the individual lymphoproliferative and myeloproliferative disorders recognized in the horse. Diagnostic techniques that are useful in cases in which hematopoietic or lymphoid tissue neoplasia are suspected include histochemical staining profiles, bone marrow aspiration, and bone marrow biopsy.


The role of chest physiotherapy in mucus hypersecretion.
Review of the data shows that chest physiotherapy (CP) is effective in clearing secretions from the lungs of patients with copious secretions (i.e., daily sputum production in excess of 30 ml). Assessment of the various components of CP shows that percussion, vibratory shaking, and breathing exercises have little to offer. Although instructed cough, the back up mucus clearance mechanism, is effective in clearing secretions its effect is less than that achieved with the forced expiration technique (FET). Postural drainage (PD) is on the whole successful in helping to drain secretions in the lungs. The administration of a beta 2-agonist via a nebulizer prior to CP in addition to promoting bronchodilation may also help to alter the physical properties of secretions, rendering them more amenable for clearance by FET/cough. Technology regarding high-frequency oscillations (HFO) has yet to be improved and its efficacy in clearing excess secretions proven prior to its inclusion in a CP treatment regimen. Intermittent positive pressure breathing (IPPB) may have a role to play in those patients where conventional CP is not possible. Data with positive expiratory pressure (PEP) are encouraging and direct evaluation is awaited regarding its effect on mucus clearance. With our present knowledge it seems reasonable to recommend that physiotherapists concentrate their CP treatment on the following: administration of a nebulized beta 2-agonist followed by PD and FET.


Homemade plasmid Miniprep solutions for affordable research in low-fund laboratories.
As a consequence of Covid-19 pandemic, the basic lab consumables are in shortage, especially in the low-income countries. Thus, the main objective of the present study is to develop and evaluate homemade solution to isolate plasmid. To pursue this objective, RNase A was overexpressed in Bl21 DE3 cells (E. coli strain) and prepared as crude refolding reaction with proper activity. Also, lysis buffers, neutralization buffer, and washing buffers were prepared. The homemade miniprep kit showed successful isolation of the px48SpCas9 plasmid. The prepared plasmid purity was enough to be used successfully in PCR amplification. In addition, to get extra benefits from this study, seven primers were designed to match the plasmid backbone to produce DNA ladder (100-1500 bp). In conclusion, we were able to have attainable working solutions for plasmid miniprep and DNA ladder.


An endogenous sleep-inducing compound is a novel competitive inhibitor of fatty acid amide hydrolase.
2-Octyl gamma-bromoacetoacetate (O gamma Br), an endogenous compound originally isolated from human cerebrospinal fluid (CSF), has previously been demonstrated to increase REM sleep duration in cats. Based on the chemical structure of O gamma Br and its reported sleep-inducing effects, we synthesized O gamma Br along with chemically related analogs and tested these compounds as inhibitors of fatty acid amide hydrolase (FAAH), a brain enzyme that degrades neuromodulatory fatty acid amides. O gamma Br was found to competitively inhibit FAAH activity with IC50 and Ki values of 2.6 microM and 0.8 microM, respectively [for the (R)-enantiomer of O gamma Br (1)]. A set of synthetic analogs of O gamma Br was examined to define the structural features required for FAAH inhibition and inhibitor potencies were assessed at pH 9.0 (near the pH optimum of FAAH) and pH 7.0. Interestingly, at pH 7.0 the gamma-halo beta-keto ester inhibitors proved to be significantly more potent than the trifluoromethyl ketone of oleic acid, one of the most potent FAAH inhibitors described to date. This study supports the possibility that O gamma Br may be a physiological regulator of FAAH activity and fatty acid amide levels in vivo. Additionally, the characterization of gamma-halo beta-keto esters as powerful FAAH inhibitors near physiological pH may aid in future studies of the enzymology and biological properties of FAAH.


Clinical and radiological results of three different techniques for fixation of the olecranon osteotomy in the surgical treatment of distal humerus fractures: A retrospective comparison of plate-screw, tension band and intramedullary nail.
The aim of this study was to compare clinical and radiological results of three different techniques (plate-screw, tension band, and intramedullary nail) in the fixation of olecranon osteotomy in patients with intercondylar fracture of the distal humerus surgically treated by the olecranon osteotomy approach. Between January 2010 and December 2018, the study was initiated with 52 patients who underwent an olecranon osteotomy approach for an AO Type C distal humerus fracture. Thirty-seven patients (19 male, 18 female) who had regular control data and a final control examination were included in the study. In osteotomy fixation, we used tension band (K wire cerclage + Screw cerclage) in 20 patients, plate-screw in eight patients, and intramedullary nail fixation in nine patients. The functional evaluation included a measurement of the range of joint motion (flexion, extension, supination, pronation); MAYO elbow performance score; disabilities of the arm, shoulder, and hand (DASH) score; and comparative grip strength measurement, and a subjective pain assessment was performed using the visual analogue scale. The mean duration of follow-up was 44 (12-84) months. The mean time to union was 14 (7-32) weeks in patients that achieved union. The mean DASH score was 22 (0-72.7), the meanMAYOelbow performance score was 84 (35-100), and the mean VAS score was 3 (1-7). The mean grip strength was 32 (8-64) kgw in the treated extremity and 37 (17-70) kgw in the intact extremity. No statistically significant difference was detected between olecranon osteotomy fixation methods (tension band, plate-screw osteosynthesis, and intramedullary nailing) in terms of union time, DASH score, MAYO scale score, VAS score, extension, supination, and pronation (P > 0.05). The average grip strength (kgw) was lower than that in the uninvolved extremity, and this ratio was statistically significant (P = 0.04). The results of this study have shown that successful and comparable radiological and functional outcomes can be obtained by all the three different olecranon osteotomy techniques in the surgical treatment of intercondylar fracture of the distal humerus. Level III, Therapeutic Study.


Synthesis and evaluation of pseudopeptide analogues of a specific CXCR4 inhibitor, T140: the insertion of an (E)-alkene dipeptide isostere into the betaII'-turn moiety.
A 14-residue peptide, T140, strongly inhibits the T-cell line-tropic HIV-1 (X4-HIV-1) infection, since this peptide functions as a specific antagonist against a chemokine receptor, CXCR4. T140 takes an antiparallel beta-sheet structure with a type II' beta-turn. In the present paper, we have designed and synthesized several T140 analogues, in which an (E)-alkene dipeptide isostere was inserted into the type II' beta-turn moiety, as a bridging study to develop nonpeptidic CXCR4 inhibitors. It has been proven that the turn region of T140 can be replaced by the above surrogate with the maintenance of strong anti-HIV activity.


Sjogren's syndrome: an update on clinical, basic and diagnostic therapeutic aspects.
The 11th International Symposium for Sjogren's syndrome was held in Athens, Greece in September 2011. This symposia is part of a long series of meetings that have attempted to meet the needs of both scientists and physicians in improving the healthcare of their patients with Sjogren's syndrome. Sjogren's syndrome affects almost 0.5% of the general population and is second only to rheumatoid arthritis amongst the systemic autoimmune diseases. More importantly, it has daily implications for the millions of sufferers around the world. The goal of this meeting, which included nearly 200 abstracts and invited lectures, was to address the critical needs in the clinical practice of Sjogren's syndrome. This volume is a composite of select papers that were presented at this meeting and attempts to provide a critical overview of clinical and basic science. The volume includes a variety of themes and, importantly, raises issues that are still unresolved but which are important in our future diagnostic and therapeutic efforts.


A Targeted Mass Spectrometry-Based Metabolomics Approach toward the Understanding of Host Responses to Huanglongbing Disease.
Candidatus Liberibacter asiaticus (CLas) is the major culprit of Huanglongbing (HLB), the most destructive citrus disease worldwide. The polymerase chain reaction (PCR) is the most common method for detecting the presence of CLas in the tree. However, due to the uneven distribution of bacteria and a minimum bacterial titer requirement, an infected tree may test false negative. Thus, our current study profiled primary and secondary metabolites of CLas-free leaves harvested from a citrus undercover protection system (CUPS) to prevent a misjudgment of CLas infection. Functional enrichment analysis revealed several metabolic pathways significantly affected by CLas infection, mainly biosynthesis of amino acids and secondary metabolites. Comparisons of CLas-infected metabolite alterations among oranges, mandarins, and grapefruits revealed that host responses to CLas were different. The metabolite signature highlighted in this study will provide a fuller understanding of how CLas bacteria affect the biosynthesis of primary and secondary metabolites in different hosts.


Does inpatient hyperglycemia predict a worse outcome in COVID-19 intensive care unit patients?
We undertook this study to evaluate the association between hyperglycemia and outcomes in patients with coronavirus disease 2019 (COVID-19) admitted to the intensive care unit (ICU). We conducted a multicenter retrospective study involving all adults with COVID-19 admitted to the ICU between March and May 2020. Patients were divided into normoglycemic (average blood glucose <140 mg/dL) and hyperglycemic (average blood glucose ≥140 mg/dL) groups. Outcomes such as mortality, need and duration of mechanical ventilation, and length of hospital and ICU stays were measured. Among 495 patients, 58.4% were male with a median age of 68 years (interquartile range [IQR]: 58.00-77.00), and baseline average blood glucose was 186.6 (SD ± 130.8). Preexisting diabetes was present in 35.8% of the studied cohort. Combined ICU and hospital mortality rates were 23.8%; mortality and mechanical ventilation rates were significantly higher in the hyperglycemic group with 31.4% vs 16.6% (P = .001) and 50.0% vs 37.2% (P = .004), respectively. Age above 60 years (hazard ratio [HR] 3.21; 95% CI 1.78, 5.78) and hyperglycemia (HR 1.79; 95% CI 1.14, 2.82) were the only significant predictors of in-hospital mortality. Increased risk for hyperglycemia was found in patients with steroid use (odds ratio [OR] 1.521; 95% CI 1.054, 2.194), triglycerides ≥150 mg/dL (OR 1.62; 95% CI 1.109, 2.379), and African American race (OR 0.79; 95% CI 0.65, 0.95). Hyperglycemia in patients with COVID-19 is significantly associated with a prolonged ICU length of stay, higher need of mechanical ventilation, and increased risk of mortality in the critical care setting. Tighter blood glucose control (≤140 mg/dL) might improve outcomes in COVID-19 critically ill patients; evidence from ongoing clinical trials is needed.


Recombinant secreted haemagglutinin protects mice against a lethal challenge of influenza virus.
Balb/c mice were immunized with 2 x 2 micrograms of purified recombinant secreted haemagglutinin, derived from the A/Victoria/3/75 (H3N2) virus. In the first immunization, Ribi adjuvant was used, while for the booster injection a monophosphoryl lipid A/muramyl dipeptide combination was chosen. Mice immunized in this way were 90-100% protected against a challenge with 20 LD50 of mouse-adapted, homologous virus (strain X47). Bromelain-solubilized haemagglutinin gave only 70% protection under comparable conditions.


Distant metastases after definitive radiotherapy for squamous cell carcinoma of the head and neck.
To analyze parameters that influence the risk of distant metastases after definitive radiotherapy. Between 1983 and 1997, 873 patients were treated with definitive radiotherapy and had follow-up for 2 years or more. Univariate and multivariate analyses were performed to evaluate risk factors that might influence the risk of distant metastases. The 5-year distant metastasis-free survival rate was 86%. Univariate analyses revealed that the risk of distant metastases was significantly influenced by gender (p =.0092), primary site (p =.0023), T stage (p <.0001), N stage (p <.0001), overall stage (p <.0001), level of nodal metastases in the neck (p <.0001), histologic differentiation (p =.0096), control above the clavicles (p <.0001), and time to locoregional recurrence (p <.0001). Multivariate analysis of freedom from distant metastases revealed that gender (p =.0390), T stage (p <.0001), N stage (p =.0060), nodal level (p <.0001), and locoregional control (p <.0001) significantly influenced this end point. Multivariate analysis revealed that gender (p =.0049), T stage (p <.0001), N stage (p <.0001), and locoregional control (p <.0001) significantly influenced cause-specific survival. The risk of distant metastases after definitive radiotherapy is 14% at 5 years and is significantly influenced by gender, T stage, N stage, nodal level, and locoregional control.


Bcl-3 deficiency protects against dextran-sodium sulphate-induced colitis in the mouse.
Bcl-3 is a member of the IκB family of proteins and is an essential negative regulator of Toll-like receptor-induced responses. Recently, a single nucleotide polymorphism associated with reduced Bcl-3 gene expression has been identified as a potential risk factor for Crohn's disease. Here we report that in contrast to the predictions of single nucleotide polymorphism (SNP) analysis, patients with Crohn's disease and ulcerative colitis demonstrate elevated Bcl-3 mRNA expression relative to healthy individuals. To explore further the potential role of Bcl-3 in inflammatory bowel disease (IBD), we used the dextran-sodium sulphate (DSS)-induced model of colitis in Bcl-3(-/-) mice. We found that Bcl-3(-/-) mice were less sensitive to DSS-induced colitis compared to wild-type controls and demonstrated no significant weight loss following treatment. Histological analysis revealed similar levels of oedema and leucocyte infiltration between DSS-treated wild-type and Bcl-3(-/-) mice, but showed that Bcl-3(-/-) mice retained colonic tissue architecture which was absent in wild-type mice following DSS treatment. Analysis of the expression of the proinflammatory cytokines interleukin (IL)-1β, tumour necrosis factor (TNF)-α and IL-6 revealed no significant differences between DSS-treated Bcl-3(-/-) and wild-type mice. Analysis of intestinal epithelial cell proliferation revealed enhanced proliferation in Bcl-3(-/-) mice, which correlated with preserved tissue architecture. Our results reveal that Bcl-3 has an important role in regulating intestinal epithelial cell proliferation and sensitivity to DSS-induced colitis which is distinct from its role as a negative regulator of inflammation.


Release of somatostatin-like immunoreactivity from the lower gut.
The present study describes the effect of the instillation of a nutrient-containing hyperosmolar volume load into the lower ileum upon plasma levels of somatostatin-like immunoreactivity. The baseline levels of SLI in the mesenteric vein were significantly higher than those in the inferior vena cava and rose significantly in both veins in response to the nutrient load. SLI levels in the venous effluent of the pancreas and stomach did not rise. Mesenteric vein SLI was similar in molecular size to the SLI in the gastric and pancreatic veins and to synthetic somatostatin. The results suggest that the lower gut may contribute to the basal circulating SLI levels and that it is released from the lower gut in response to a nutrient-containing hyperosmolar volume load.


Familial hypertryptophanemia, tryptophanuria and indoleketonuria.
Two patients presenting with mental subnormality and severe emotional lability were examined for possible inborn errors of metabolism. Amino acid and organic acid analyses of their plasma and urine revealed a novel and grossly abnormal metabolism of tryptophan. Basal levels of plasma tryptophan were ten times that of controls and the 24-hour urinary excretion of tryptophan was up to 50 times greater than normal. Both patients excreted about 100 times the normal amounts of indoleacetic, indolelactic and indolepyruvic acids. Administration of oral antibiotic to one of the patients to eliminate intestinal bacteria did not result in any reduction in the excretion of the indoleic acids. The results are interpreted in terms of a possible congenital defect in the normal conversion of tryptophan to kynurenine.


Bromocriptine treatment for cocaine addiction: association with plasma prolactin levels.
Bromocriptine is a dopamine receptor agonist used with mixed success in the treatment of cocaine addiction. Variations in dopamine receptor sensitivity may help account for these differences. We evaluated this question in a 24-week outpatient controlled clinical trial in 70 cocaine-abusing (DSM-III) men (86% African-American, mean age 34 years, mean 39 months of regular cocaine use [predominantly smoked]). Subjects received 4 weeks of inpatient treatment. During the last 2 weeks they were inducted onto bromocriptine (maximum dose 2.5mg po tid) (n=35) or placebo (n=35). Plasma prolactin concentrations were assayed before and after the first bromocriptine dose (1.25mg po) as a measure of dopamine receptor sensitivity. After discharge, subjects continued on medication with weekly group counseling. Bromocriptine significantly suppressed prolactin concentrations (4.4 ng/ml decrease), while placebo did not (0.1 ng/ml decrease). Both groups decreased their cocaine use, with no significant group differences in retention in treatment or proportion of cocaine-positive urine samples. There was no significant association between basal plasma prolactin concentrations or prolactin response to first bromocriptine dose and either outcome measure. These data do not support the efficacy of bromocriptine treatment nor a role for prolactin concentration in predicting treatment response.


Genetic Control of Potassium Channels.
Approximately 80 genes in the human genome code for pore-forming subunits of potassium (K(+)) channels. Rare variants (mutations) in K(+) channel-encoding genes may cause heritable arrhythmia syndromes. Not all rare variants in K(+) channel-encoding genes are necessarily disease-causing mutations. Common variants in K(+) channel-encoding genes are increasingly recognized as modifiers of phenotype in heritable arrhythmia syndromes and in the general population. Although difficult, distinguishing pathogenic variants from benign variants is of utmost importance to avoid false designations of genetic variants as disease-causing mutations.


Polydopamine: surface coating, molecular imprinting, and electrochemistry-successful applications and future perspectives in (bio)analysis.
Dopamine oxidation and self-polymerization have recently attracted great interest arising from the versatile chemistry of this endogenous catecholamine. Particularly interesting are the applications of polydopamine for surface coating, molecular imprinting, and electrochemistry, which are reviewed here, covering the broad fields of medicine, materials science, and (bio)analytical chemistry. Nonetheless, the peculiar physicochemical properties of dopamine and polydopamine, due to the redox potential of the catechol moiety, are not fully exploited. We have confidence in increasing the applications of dopamine through a large variety of research approaches, including the use of naturally occurring or synthetic dopamine analogues and copolymers. Accordingly, our efforts in this direction are focused on proposing a role for polydopamine in quantitative applications, evaluating analytical performance, cost, reproducibility, and versatility of the methods developed, and also revisiting standard (bio)analytical platforms.


Eosinophils induces glioblastoma cell suppression and apoptosis - Roles of GM-CSF and cysteinyl-leukotrienes.
Glioblastoma is the most common and lethal primary brain tumor in adults. Despite the available cancer treatments, the recurrence of the tumor is high, and the survival rate is low. New approaches to antitumor therapies are needed. Eosinophils are prominent in allergic diseases and accumulate in several human brain tumors. Recently, the antitumor role of eosinophils has been targeted as eosinophils release several cytotoxic factors that induce cell impairment and death. Here we aim to evaluate the interaction of the eosinophil and glioblastoma cells, the mechanism involved in the potential killing of the glioblastoma cells by the eosinophils, and how allergy/asthma could confer a better glioblastoma prognosis. Eosinophils and serum from asthmatic and non-asthmatic donors were cultivated with different glioblastoma cell lines. Glioblastoma cells recruit eosinophils via GM-CSF signaling, activating and increasing eosinophil survivability and function on a GM-CSF-dependent manner. Eosinophils reduce glioblastoma cells metabolism, proliferation, and migration, via Fas/FasL. Cysteinyl-leukotrienes are accounted for the asthmatic serum enhancement of the glioblastoma cell migration and proliferation. Cysteinyl-leukotrienes enhance glioblastoma cell proliferation and migration, albeit activate eosinophils that suppress glioblastoma cells. Eosinophils have the potential to be key cells on glioblastoma therapeutics, as allergy and eosinophilia are correlated with a better glioblastoma prognosis. Eosinophils are elicited and attach to glioblastoma cells, where, by its cytotoxic function, via Fas/FasL, hind glioblastoma cell metabolism, proliferation, migration, and induce cell death.


The Clinical Utility of Next Generation Sequencing Results in a Community-Based Hereditary Cancer Risk Program.
Since the 2013 Supreme Court ruling on BRCA1/BRCA2 patenting, hereditary cancer gene panels now include BRCA1 and BRCA2, making these panels an option for first-tier testing. However, questions remain about the clinical utility and implications of these panels for medical management with inclusion of genes of unknown to moderate penetrance. To better understand how use of these panels affected our practice, we reviewed patients who underwent testing in our clinic from July 1, 2013 through May 23, 2014. Indications for testing included personal and/or family history of breast and/or ovarian cancer. A total of 136 patients underwent panel testing via a single commercial laboratory; 12 (8.8 %) patients were positive for a pathogenic or likely pathogenic mutation (four BRCA2 mutations, two TP53 mutations, one CDH1 mutation, two ATM mutations, and one patient each with a CHEK2, NBN, or PALB2 mutation). Of these positive patients, 100 % met the National Comprehensive Cancer Network (NCCN) guidelines for Hereditary Breast and Ovarian Cancer genetic testing (2.2014). Mutations in seven of twelve (58 %) patients led to changes in medical management; three of seven (43 %) had a non-BRCA1 or BRCA2 gene mutation. Our findings suggest that there is clinical utility of panels that include genes of unknown to moderate penetrance.


Splenic cysts: aspiration, sclerosis, or resection.
Percutaneous aspiration and tetracycline sclerosis is a safe but temporary therapy of large splenic cysts in children. Between 1985 and 1987, three girls with splenic cysts were seen. Their ages ranged from 5 to 14 years, and the cysts were from 8 to 16 cm in diameter. Despite their large size, all were asymptomatic and were discovered upon physical examination or ultrasound for unrelated conditions. All cysts were avascular by scan and had irregular crenated or smooth walls by ultrasound. Further investigation excluded infectious or parasitic causes. Each cyst was aspirated for diagnosis, and a pigtail catheter was inserted for drainage and sclerotherapy. All needle aspirations resulted in cyst collapse, but in one patient the pigtail catheter insertion was unsuccessful, and in the other two cases, multiple attempts of tetracycline sclerosis failed to obliterate the cysts. There were no other complications. Surgery for the recurrent splenic cysts was performed 3 months to 2 years following the percutaneous procedures. The two patients operated on with 3 months of aspiration underwent successful partial splenectomy and have normal splenic function by ultrasound scan, and absence of RBCs. The third patient had progression of the cystic disease throughout the spleen, and required splenectomy. Pathology confirmed multiseptate congenital mesothelial cysts in the first two patients and massive lymphangiomatosis in the third. In all three cases, percutaneous therapy was safe but did not result in long-term control. In one patient, the cystic disease progressed following sclerotherapy and may have influenced the need for complete splenectomy. Prior manipulation did not adversely affect the dissection and mobilization of the spleens.(ABSTRACT TRUNCATED AT 250 WORDS)


Detection of IgM and IgG antibodies to Chlamydophila pneumoniae in pediatric community-acquired lower respiratory tract infections.
Chlamydophila pneumoniae (C. pneumoniae) is an emerging infectious agent with a spectrum of clinical manifestations including lower and upper respiratory tract infections. To investigate the role of C. pneumoniae in community-acquired lower respiratory tract infections (LRTIs) in children using serological tests. Two hundred children, age 2 months to 12 years, hospitalized for community-acquired LRTIs were investigated for C. pneumoniae etiology. We investigated 200 children hospitalized for community-acquired LRTIs, using ELISA for detecting anti-C. pneumoniae IgM and IgG antibodies. The demographic, clinical and radiological findings for C. pneumoniae antibody positive and C. pneumoniae antibody negative cases were compared. Data analysis was performed by Chi-square test and Fisher's exact tests using Epi Info (2002). Clinical and radiological findings in both the groups were comparable. Serological evidence of C. pneumoniae infection was observed in 12 (6%) patients; specific IgM antibodies were detected in 11 (91.67%; specific IgG antibodies in 1 (8.33%) patients, while 4-fold rise in C. pneumoniae IgG antibody titers were noted in none of the patients. C. pneumoniae has a role in community-acquired LRTIs, even in children aged < 5 years. Serological detection using ELISA would enable pediatricians in better management of C. pneumoniae infections.


Dengue infections during pregnancy: a case series from Sri Lanka and review of the literature.
Dengue is a major public health problem in several countries. Few reports document presentations and outcomes of dengue during pregnancy. In many endemic countries as the average age of getting dengue infections is shifting upwards, dengue in pregnancy is likely to be encountered more frequently. Patterns of dengue in patients from different regions are needed if we are to draft evidence based management guidelines. To document clinical and laboratory findings in a cohort of hospitalised patients with dengue during pregnancy in Sri Lanka and compare their presentation and outcomes with previously published cases. Clinical, laboratory, maternal and fetal outcomes and demographic information were collected from patients with confirmed dengue infections during pregnancy treated in a Maternity Hospital in Sri Lanka from 1 January 2000 to 30 June 2004. The Medline database was searched to identify reports relating to dengue infection during pregnancy since 1965. Twenty-six patients [mean (S.D.) age: 29 (4.2) years] were studied. One (3.8%), 2 (7.7%) and 20 (77%) presented in the first, second and third trimesters of pregnancy, and 3 (11.5%) in the immediate post-partum period. Seventeen (65.3%) had primary and nine (34.7%) secondary dengue infections. Ten (38.5%) had DF, 6 (23.1%): DHF grade I and 10 (38.5%): DHF grade II. Five (19.2%) and three (11.5%) patients who first presented with cough/breathlessness or vaginal bleeding, were initially managed as having a pulmonary embolism or a primary obstetric cause for their vaginal bleeding. Bradycardia was noted in three of the four patients who had a cardiac arrhythmia. Seven (26.9%) needed admission to an ICU. Raised AST and ALT levels were seen in 81.2% and 43.7% of 16 patients in whom liver function tests results were available. No fetal malformations were seen in any of the babies born. The single patient who developed DHF in the first trimester had an abortion while having acute symptoms of dengue. Awareness of clinical and laboratory manifestations of dengue in pregnancy should allow its early recognition and institution of appropriate treatment. Reports on dengue in pregnancy from different regions should allow more evidence-based guidelines to be formulated for optimum evaluation and care of such patients. Our data contributes towards this goal.


LncRNA KCNA2-AS regulates spinal astrocyte activation through STAT3 to affect postherpetic neuralgia.
Postherpetic neuralgia (PHN) is the most common complication of herpes zoster, but the mechanism of PHN is still unclear. Activation of spinal astrocytes is involved in PHN. Our study aims to explore whether lncRNA KCNA2 antisense RNA (KCNA2-AS) regulates spinal astrocytes in PHN through signal transducers and activators of transcription 3 (STAT3). Varicella zoster virus (VZV)-infected CV-1 cells were injected into rats to construct a PHN model. Primary spinal cord astrocytes were activated using S-Nitrosoglutathione (GSNO). Glial fibrillary acidic protein (GFAP; marker of astrocyte activation), phosphorylated STAT3 (pSTAT3), and KCNA2-AS were analyzed by immunofluorescence and RNA fluorescence in situ hybridization. RNA pull-down and RNA immunoprecipitation were used to detect binding of KCNA2-AS to pSTAT3. KCNA2-AS was highly expressed in the spinal cord tissue of PHN model rats, and was positively correlated with GFAP expression. GFAP was significantly increased in GSNO-induced cells, but the knockdown of KCNA2-AS reversed this result. Meanwhile, pSTAT3 was significantly increased in GSNO-induced cells, but knockdown of KCNA2-AS reduced pSTAT3 within the nucleus while the total pSTAT3 did not change significantly. pSTAT3 bound to KCNA2-AS and this binding increased with GSNO treatment. Furthermore, knockdown of KCNA2-AS in PHN model rats relieved mechanical allodynia. Down-regulation of KCNA2-AS alleviates PHN partly by reducing the translocation of pSTAT3 cytoplasm to the nucleus and then inhibiting the activation of spinal astrocytes.


Shedding of Marburg Virus in Naturally Infected Egyptian Rousette Bats, South Africa, 2017.
We detected Marburg virus RNA in rectal swab samples from Egyptian rousette bats in South Africa in 2017. This finding signifies that fecal contamination of natural bat habitats is a potential source of infection for humans. Identified genetic sequences are closely related to Ravn virus, implying wider distribution of Marburg virus in Africa.


Echo-Guided Pericardiocentesis: When and How Should It Be Performed?
Abnormal accumulation of pericardial fluid is a common cardiac condition with different etiologies. Draining of the pericardial fluid (pericardiocentesis) is often indicated for diagnostic and therapeutic purposes and is performed in an elective or emergent setting. Echocardiography is the primary imaging method for diagnosing, localizing, and quantifying pericardial effusion as well as evaluating its hemodynamic effects, including the presence of cardiac tamponade. In this manuscript, we review the indications for pericardiocentesis and provide practical step-by-step guidance for echo-guided pericardiocentesis. Echo-guidance is an effective method to improve the safety and efficacy of pericardiocentesis. In experienced hands and with a stepwise approach, procedural outcomes are excellent, and complication rates are very low. Asymptomatic small idiopathic effusions have a benign course and can be left untreated. Prolonged drainage with an indwelling pericardial catheter is key for preventing fluid re-accumulation, and the use of colchicine to prevent fluid recurrence is encouraged whenever possible. Understanding how to evaluate the significance of a pericardial effusion as well as the procedural steps in the performance of a pericardiocentesis are essential for optimal outcomes in treating patients with pericardial effusions and tamponade.


Oscillatory cluster patterns in a homogeneous chemical system with global feedback
Oscillatory clusters are sets of domains in which nearly all elements in a given domain oscillate with the same amplitude and phase. They play an important role in understanding coupled neuron systems. In the simplest case, a system consists of two clusters that oscillate in antiphase and can each occupy multiple fixed spatial domains. Examples of cluster behaviour in extended chemical systems are rare, but have been shown to resemble standing waves, except that they lack a characteristic wavelength. Here we report the observation of so-called 'localized clusters'--periodic antiphase oscillations in one part of the medium, while the remainder appears uniform--in the Belousov-Zhabotinsky reaction-diffusion system with photochemical global feedback. We also observe standing clusters with fixed spatial domains that oscillate periodically in time and occupy the entire medium, and irregular clusters with no periodicity in either space or time, with standing clusters transforming into irregular clusters and then into localized clusters as the strength of the global negative feedback is gradually increased. By incorporating the effects of global feedback into a model of the reaction, we are able to simulate successfully the experimental data.


Organ-specific, integrated omics data-based study on the metabolic pathways of the medicinal plant Bletilla striata (Orchidaceae).
Bletilla striata is one of the important species belonging to the Bletilla genus of Orchidaceae. Since its extracts have an astringent effect on human tissues, B. striata is widely used for hemostasis and healing. Recently, some other beneficial effects have also been uncovered, such as antioxidation, antiinflammation, antifibrotic, and immunomodulatory activities. As a key step towards a thorough understanding on the medicinal ingredient production in B. striata, deciphering the regulatory codes of the metabolic pathways becomes a major task. In this study, three organs (roots, tubers and leaves) of B. striata were analyzed by integrating transcriptome sequencing and untargeted metabolic profiling data. Five different metabolic pathways, involved in polysaccharide, sterol, flavonoid, terpenoid and alkaloid biosynthesis, were investigated respectively. For each pathway, the expression patterns of the enzyme-coding genes and the accumulation levels of the metabolic intermediates were presented in an organ-specific way. Furthermore, the relationships between enzyme activities and the levels of the related metabolites were partially inferred. Within the biosynthetic pathways of polysaccharides and flavonoids, long-range phytochemical transportation was proposed for certain metabolic intermediates and/or the enzymes. The data presented by this work could strengthen the molecular basis for further studies on breeding and medicinal uses of B. striata.


A Beclin 1-targeting stapled peptide synergizes with erlotinib to potently inhibit proliferation of non-small-cell lung cancer cells.
Epidermal Growth Factor Receptor (EGFR) is a major drug target for non-small-cell lung carcinoma (NSCLC). Tyrosine Kinase Inhibitors (TKIs) like erlotinib are potent inhibitors of EGFR and have achieved impressive clinical success against NSCLC. However, NSCLC cells readily develop resistance to TKIs by acquiring mutations in EGFR or other oncogenes. Novel strategies to inhibit EGFR are needed to overcome this urgent problem of TKI resistance. Beclin 1 is an essential autophagy protein and is intimately involved in tumorigenesis and EGFR signaling. Here we present data to show that a Beclin 1-targeting stapled peptide Tat-SP4 can inhibit the EGFR signaling pathway by enhancing the Beclin 1-mediated endolysosomal degradation of EGFR. This inhibition mechanism is orthogonal to that employed by TKIs and is effective against either wild-type or mutant EGFR. Tat-SP4 alone showed moderate anti-proliferative efficacy in NSCLC cells but synergized with erlotinib to potently inhibit NSCLC proliferation. These results suggest that Beclin 1-targeting stapled peptides may be used in combination with TKIs to enhance their efficacy, particularly for NSCLC subtypes refractory to current regiments.


Sarcoidosis.
(1) Sarcoidosis may affect any portion of the visual system. (2) Eye involvement may occur prior to, concurrent with, or after systemic manifestations. (3) Ocular manifestations may be the only complaint. (4) No age group is exempt. (5) Diagnosis may require a high index of suspicion. (6) Ocular sarcoidosis may be more common than is usually appreciated. (7) Ocular manifestations may be more common in black and oriental patients with sarcoidosis than in whites.


PD-1/PD-L1 inhibitors in treatment-naïve, advanced non-small cell lung cancer patients with < 1% PD-L1 expression: a meta-analysis of randomized controlled trials.
PD-1/PD-L1 inhibitors prolong survival in treatment-naïve, locally advanced, and metastatic non-small cell lung cancer (NSCLC) with positive PD-L1 expression (> 1%/ > 50%). Recent evidence has suggested that tumors with < 1% PD-L1 expression may also be predictive of PD-1/PD-L1 inhibiting agents. Systematic review and meta-analysis were conducted of randomized controlled trials (RCTs) evaluating PD-1/PD-L1 inhibitors that have assessed tumors with < 1% PD-L1 expression (negative PD-L1 expression). PD-1/PD-L1 inhibitors-chemotherapy combinations (PC) were compared with histology-selected chemotherapy with respect to overall survival (OS) and progression-free survival (PFS). Twelve RCTs comprising 5410 participants (PD-1/PD-L1 inhibitors-chemotherapy: 3051; chemotherapy: 2359) met the inclusion criteria. Tumors with PD-L1 expression < 1% were evident in 38.9% of the pooled study population. A significant OS [hazard ratio (HR) 0.71 95% confidence interval (CI) 0.63-0.80, p < 0.00001] and PFS [HR 0.65 95% CI 0.58-0.72, p < 0.00001] benefit of PC was evident in tumors with negative PD-L1 expression. PD-1/PD-L1 inhibitors-chemotherapy combinations were more likely to achieve an objective response than chemotherapy [odds ratio, 1.86; 95% CI, 1.46-2.38, p < 0.00001]. Histologic subtypes and diagnostic assays did not modify the OS and PFS treatment effects for PC compared to chemotherapy. Tumors harboring < 1% PD-L1 expression are likely to benefit from PD-1/PD-L1 inhibitor-chemotherapy regimens in advanced NSCLC.


Interactions effects of nano-microplastics and heavy metals in hybrid snakehead (Channa maculata ♀ × Channa argus ♂).
The interaction between microplastics and contaminants has potentially generated new undefined risks on animals and ecosystems, and nano-microplastics are considered to be more harmful than microplastics. This experiment investigated the interactions and effects of nano-microplastics with heavy metals cadmium in hybrid snakehead. Different concentrations of nano-microplastics 80 nm (50 μg/L and 500 μg/L) and Cd (50 μg/L) were used for exposure, and four sampling points were set for 24 h, 48 h, 96 h and clear-48 h. Results indicated that the morphology of gill was altered under the influence of nano-microplastics and cadmium, and the damage was aggravated with time. Nano-microplastics and Cd can cause oxidative damage to fish liver partly by effect the activities of antioxidant enzyme, and significantly suppressed the expressions of genes related to the inflammation (IL-1β and TNF-α) and as well as significantly up-regulated the expression of genes HSP70 and SOD. Additionally, the mRNA levels of MT gene can be speculated that the heavy metal cadmium may accumulated in the body over time. And the concentration of heavy metals will also affect their accumulation in the body. Our study elucidated the nano-microplastics and Cd will increase the impact on environmental and organisms that the nano-microplastics contribute to the bioaccumulation of metals, which served as a new support for study the interaction between environmental contaminants.


Can proton pump inhibitors reduce rebleeding following Histoacryl sclerotherapy for gastric variceal hemorrhage?
To evaluate the efficacy of proton pump inhibitors (PPIs) in reducing rebleeding and bleeding-related death rates after endoscopic gastric variceal obliteration (GVO) using N-butyl-2-cyanoacrylate (NBC). This study enrolled 341 patients who were consecutively diagnosed with and treated for bleeding gastric varices. The patients were divided into PPI and non-PPI groups, and their endoscopic findings, initial hemostasis outcomes, rebleeding and bleeding-related death rates, and treatment-related complications were analyzed. The rate of initial hemostasis was 97.1%. rebleeding occurred in 2.2% of patients within 2 weeks, 3.9% of patients within 4 weeks, 18.9% of patients within 6 months, and 27.6% of patients within 12 months of the GVO procedure. A previous history of variceal bleeding (relative risk [RR], 1.955; 95% confidence interval [CI], 1.263 to 3.028; p = 0.003) and use of PPIs (RR, 0.554; 95% CI, 0.352 to 0.873; p = 0.011) were associated with rebleeding. Child-Pugh class C (RR, 10.914; 95% CI, 4.032 to 29.541; p < 0.001), failure of initial hemostasis (RR, 13.329; 95% CI, 2.795 to 63.556; p = 0.001), and the presence of red-colored concomitant esophageal varices (RR, 4.096; 95% CI, 1.320 to 12.713; p = 0.015) were associated with bleeding-related death. The prophylactic use of PPIs reduces rebleeding after GVO using NBC in patients with gastric variceal hemorrhage. However, prophylactic use of PPIs does not reduce bleeding-related death.


Chiral tertiary 2-furyl alcohols: diversified key intermediates to bioactive compounds. Their enantioselective synthesis via (2-furyl)aluminium addition to ketones catalyzed by a titanium catalyst of (S)-BINOL.
Novel asymmetric 2-furyl additions of (2-furyl)AlEt(2)(THF) to aromatic ketones and one alpha,beta-unsaturated ketone catalyzed by a titanium catalyst of 10-20 mol% (S)-BINOL are reported to furnish tertiary furyl alcohols in good to excellent enantioselectivities of 87-93% ee.


Purification, structure and properties of the respiratory nitrate reductase of Klebsiella aerogenes.
1. The respiratory nitrate reductase of Klebsiella aerogenes was solubilized from the bacterial membranes by deoxycholate and purified further by means of gel chromatography in the presence of deoxycholate, and anion-exchange chromatography. 2. Dependent on the isolation procedure two different homogeneous forms of the enzyme, having different subunit compositions, can be obtained. These forms are designated nitrate reductase I and nitrate reductase II. Both enzyme preparations are isolated as tetramers having sedimentation constants (s20,w) of 22.1 S and 21.7 S for nitrate reductase I and II, respectively. The nitrate reductase I tetramer has a molecular weight of about 106. 3. In the presence of deoxycholate both enzyme preparations dissociate reversibly into their respective monomeric forms. The monomeric form of nitrate reductase I has a molecular weight of about 260 000 and a sedimentation constant of 9.8 S. For nitrate reductase II these values are 180 000 and 8.5 S, respectively. 4. Nitrate reductase I consists of three different subunits, having molecular weights of 117 000; 57 000 and 52 000, which are present in a 1:1:2 molar ratio, respectively. Nitrate reductase II contains only the subunits with a molecular weight of 117 000 and 57 000 in a equimolar ratio. 5. Treatment at pH 9.5 in the presence of deoxycholate and 0.05 M NaCl or ageing removes the 52 000 Mr subunit from nitrate reductase I. This smallest subunit, in contrast to the other subunits, is a basic protein. 6. The 52 000 Mr subunit has no catalytic function in the intramolecular electron transfer from reduced benzylviologen to nitrate. However, it appears to have a structural function since nitrate reductase II, which lacks this subunit, is much more labile than nitrate reductase I. Inactivation of nitrate reductase II can be prevented by the presence of deoxycholate. 7. The spectrum of the enzyme resembles that of iron-sulfur proteins. No cytochromes or contaminating enzyme activities are present in the purified enzyme. Only reduced benzylviologen was found to be capable of acting as an electron donor. 8. p-Chlormercuribenzoate enhances the enzymatic activity at concentrations of 0.1 mM and lower. At higher p-chlormercuribenzoate concentrations the enzymatic activity is inhibited non-competitively with either nitrate or benzylviologen as a substrate. The inhibition is not counteracted by cysteine.


Early cases of SARS-CoV-2 infection in Uganda: epidemiology and lessons learned from risk-based testing approaches - March-April 2020.
On March 13, 2020, Uganda instituted COVID-19 symptom screening at its international airport, isolation and SARS-CoV-2 testing for symptomatic persons, and mandatory 14-day quarantine and testing of persons traveling through or from high-risk countries. On March 21, 2020, Uganda reported its first SARS-CoV-2 infection in a symptomatic traveler from Dubai. By April 12, 2020, 54 cases and 1257 contacts were identified. We describe the epidemiological, clinical, and transmission characteristics of these cases. A confirmed case was laboratory-confirmed SARS-CoV-2 infection during March 21-April 12, 2020 in a resident of or traveler to Uganda. We reviewed case-person files and interviewed case-persons at isolation centers. We identified infected contacts from contact tracing records. Mean case-person age was 35 (±16) years; 34 (63%) were male. Forty-five (83%) had recently traveled internationally ('imported cases'), five (9.3%) were known contacts of travelers, and four (7.4%) were community cases. Of the 45 imported cases, only one (2.2%) was symptomatic at entry. Among all case-persons, 29 (54%) were symptomatic at testing and five (9.3%) were pre-symptomatic. Among the 34 (63%) case-persons who were ever symptomatic, all had mild disease: 16 (47%) had fever, 13 (38%) reported headache, and 10 (29%) reported cough. Fifteen (28%) case-persons had underlying conditions, including three persons with HIV. An average of 31 contacts (range, 4-130) were identified per case-person. Five (10%) case-persons, all symptomatic, infected one contact each. The first 54 case-persons with SARS-CoV-2 infection in Uganda primarily comprised incoming air travelers with asymptomatic or mild disease. Disease would likely not have been detected in these persons without the targeted testing interventions implemented in Uganda. Transmission was low among symptomatic persons and nonexistent from asymptomatic persons. Routine, systematic screening of travelers and at-risk persons, and thorough contact tracing will be needed for Uganda to maintain epidemic control.


Molecular dynamics simulations of the interaction of glucose with imidazole in aqueous solution.
Molecular dynamics simulations were carried out on a concentrated system of β-d-glucopyranose and imidazole molecules in a periodic box of water at 298K. The purpose of the simulations was to determine whether or not there was any tendency for these two solutes to associate in an aqueous environment, as has previously been observed for other planar functional groups from amino acid side chains, such as the indole group of tryptophan or the phenolic group of tyrosine. A weak stacking interaction between β-d-glucopyranose, as a model for cellulose, and imidazole was indeed observed, with an energy of ∼0.25kcal/mol per pair, less than kT. Somewhat surprisingly, considerable imidazole self-association into small aggregates (dimers and trimers) was also observed, with binding energies of ∼0.4kcal/mol per pair, although still less than kT. Considerable non-stacked interactions between glucose and imidazole through hydrogen bonding were also found. These hydrogen bonds primarily involved the N3 atom of imidazole, because the N-H group of N1 was insufficiently polar to compete for water hydrogen bond partners.


Evaluating Health-Related Quality of Life and School Attendance in a Multidisciplinary School Program for Youth with Significant Medical Needs.
Many youth with significant medical needs have difficulty attending traditional schools due to academic, physical, and psychosocial challenges. To meet the needs of these youth and prepare them for a successful transition to a traditional school, a multidisciplinary school program (MSP) provides support in these three domains. The aims of this program evaluation are to describe the operation of the novel MSP, characterize participants, and determine the impact of participation as related to school attendance and health-related quality of life (HRQoL). Attendance in the MSP was significantly higher than school attendance estimates provided by caregivers prior to participation in the program. Youth reported significant improvement in physical functioning and total HRQoL. Caregivers reported significant improvement in academic functioning and total HRQoL of youth. The MSP represents a unique educational model for youth with significant medical issues that also provides physical and psychosocial support. Initial findings highlight the potential positive impact of this model for this population of youth.


New-born infants with severe hyaline membrane disease: radiological evaluation during high frequency oscillatory versus conventional ventilation.
The aim of our study was to determine the impact of treatment with exogenous surfactant (ES) and high frequency oscillatory ventilation (HFOV) on the radiological appearance and clinical course of hyaline membrane disease (HMD) in new-born infants. New-born infants (18) (median weight, 1010 g) with severe HMD (stages 3.5 and 4) who were treated with ES and HFOV were matched by birth weight and severity of disease with 18 new-born infants treated with ES and conventional mechanical ventilation (CV). Chest radiograms taken on days 1, 2/3, 4/5, 7, 14 and 28 were analyzed to check for the severity of generalized parenchymal opacities (GPO), local opacifications, pulmonary interstitial emphysema (PIE), gross air leak, general and localized overinflation, bronchopulmonary dysplasia (BPD) and clinical variables such as survival rates, duration of mechanical ventilation, mean airway pressure and inspired oxygen concentration. At 4 weeks of age, new-born infants treated by HFOV had less severe GPO (median degree 1.5 vs. 3), less PIE (1 vs. 7 patients) and fewer signs of BPD (median BPD degree 1.5 vs. 2.6). The incidence of pneumothorax and of local opacifications were similar in both groups. New-born infants on HFOV had a lower mortality rate (5 vs. 13), needed fewer days of mechanical ventilation (median 15 vs. 23 days) and lower inspiratory oxygen concentrations (median FiO2 0.38 vs. 0.64). In new-born infants with HMD, treatment with ES and HFOV resulted in a favourable radiological and clinical outcome as compared to treatment with ES and CV.


Assessment of Patient-Specific Human Leukocyte Antigen Genomic Loss at Relapse After Antithymocyte Globulin-Based T-Cell-Replete Haploidentical Hematopoietic Stem Cell Transplant.
Patient-specific human leukocyte antigen (HLA) genomic loss (HLA loss) is one of the reputed mechanisms of leukemia immune escape and relapse after haploidentical hematopoietic stem cell transplant (HSCT). However, clinical characteristics and prognosis of this distinct relapse type in the setting of haploidentical HSCT based on antithymocyte globulin (ATG) T-cell-replete conditioning are still unknown, especially for patients with lymphoid leukemia. To identify the incidence of and patient characteristics associated with HLA loss at hematologic cancer relapse after ATG-based haploidentical HSCT and to assess overall survival after HLA loss at relapse. This retrospective and multicenter case series study used data from medical records to identify patients who experienced relapse of hematologic cancer after receipt of ATG-based haploidentical HSCT. The study included 788 consecutive patients aged 8 to 70 years with lymphoid or myeloid leukemia who received ATG T-cell-replete haploidentical HSCT at the Zhejiang Cooperative Group for Blood and Marrow Transplantation between May 1, 2012, and May 31, 2021. Relapse after ATG-based haploidentical HSCT. Incidence, risk factors, and postrelapse overall survival among patients with HLA loss at hematologic cancer relapse after receipt of haploidentical HSCT. Logistic regression analysis was used to identify characteristics associated with the likelihood of HLA loss, and Kaplan-Meier and Cox regression analyses were performed to evaluate postrelapse survival. A total of 788 patients who received haploidentical HSCT for hematologic cancer were identified, 180 of whom experienced relapse after HSCT. Of those, 106 evaluable patients (median age, 30.9 years [range, 8.3-64.6 years]; 54 female [50.9%] and 52 male [49.1%]) were screened for HLA loss, which was detected in 54 patients (50.9%). Patients with HLA loss experienced relapse later than those without HLA loss (lymphoid group: median, 323 days [range, 98-2056 days] vs 151 days [range, 57-2544 days]; P = .01; myeloid group: median, 321 days [range, 55-1574 days] vs 223 days [range, 68-546 days]; P = .03). Among patients with lymphoid leukemia, those with minimal residual disease positivity before hematologic relapse (odds ratio [OR], 28.47; 95% CI, 1.99-407.98; P = .01), those with chronic graft-vs-host disease (OR, 27.68; 95% CI, 1.40-546.88; P = .03), and those with more than 180 days between HSCT and relapse (OR, 6.91; 95% CI, 1.32-36.22; P = .02) were more likely to lose unshared HLA at relapse, whereas male patients (OR, 0.03; 95% CI, 0.003-0.32; P = .04) were more likely to preserve their HLA genome at relapse. Patients with myeloid leukemia had different factors associated with HLA loss, including underweight status (OR, 0.10; 95% CI, 0.02-0.60; P = .01) and acute graft-vs-host disease (OR, 4.84; 95% CI, 1.14-20.53; P = .03). The receipt of preemptive donor lymphocyte infusion among patients with minimal residual disease recurrence did not postpone hematologic cancer relapse in those with HLA loss (median, 322 days [range, 204-1030 days]) compared with no receipt of donor lymphocyte infusion (median, 340 days [range, 215 days to not available]; P > .99). In this study, HLA loss at leukemia relapse occurred frequently after receipt of ATG-based haploidentical HSCT. The identification of risk factors associated with HLA loss would help to prompt screening, avoid potentially harmful infusions of donor T cells, and develop alternative therapeutic strategies.


Trapping of wide range mass-to-charge ions and dependence on matrix amount in internal source MALDI-FTMS.
Internal ionization source MALDI-FTMS shows clear variation of number average molecular weight (M(n)) for an equimolar mixture of four PEG polymers (PEG 2000, PEG 4000, PEG 6000, PEG 8000) when the 2,5-dihydroxy benzoic acid (DHB) matrix to PEG ratio is varied or the laser power is changed. As the matrix to analyte ratio is increased, the analyte signal of higher molecular mass ions increases and the signal of lower mass ions decreases. Laser power dependence studies show a similar trend. Possible explanations for these observations are discussed.


Kisspeptin and its receptor: new gatekeepers of puberty.
The recent finding that the hormone kisspeptin plays a pivotal role in the onset of puberty is one of the biggest discoveries in human reproductive biology in 30 years. Mutations in the receptor for kisspeptin cause humans and mice to fail to reach puberty and to be sterile. It is the first time since the identification of gonadotrophin-releasing hormone that a single gene is found to have such a dramatic effect on reproduction. This discovery opens new possibilities in the treatment of reproductive disorders such as delayed or advanced puberty, infertility and sex hormone-dependent cancers.


Comparison of in-house biotin-avidin tetanus IgG enzyme-linked-immunosorbent assay (ELISA) with gold standard in vivo mouse neutralization test for the detection of low level antibodies.
Detection of anti-tetanus antibody levels is necessary for both determination of the immune status of individuals and also for planning preventive measures. ELISA is the preferred test among in vitro tests however it can be affected by the cross reacting antibodies. A previously developed in-house ELISA test was found not reliable for the antibody levels ≤1.0IU/ml. A new method was developed to detect low antibody levels correctly. The aim of the present study was to compare the results of the newly developed in-house biotin-avidin tetanus IgG ELISA test with the in vivo mouse neutralization test, for the antibody levels ≤1.0IU/ml. A total of 54 serum samples with the antibody levels of three different levels, =0.01IU/ml, 0.01-0.1IU/ml, 0.1-1IU/ml, which were detected by in vivo mouse neutralization test were studied by the newly developed in-house biotin-avidin tetanus IgG ELISA test. Test was validated by using five different concentrations (0.01IU/ml, 0.06IU/ml, 0.2IU/ml, 0.5IU/ml, 1.0IU/ml). A statistically significant correlation (r2=0.9967 p=0,001) between in vivo mouse neutralization test and in-house biotin-avidin tetanus IgG ELISA test, was observed. For the tested concentrations intra-assay, inter-assay, accuracy, sensitivity, specificity and coefficients of variations were determined as ≤15%. In-house biotin-avidin tetanus IgG ELISA test can be an alternative method to in vivo mouse neutralization method for the detection of levels ≤1.0IU/ml. By using in-house biotin-avidin tetanus IgG ELISA test, individuals with non protective levels, will be reliably detected.


Contrast-enhanced magnetic resonance angiography (CE-MRA) of intra- and extra-cranial vessels in dogs.
Until recently intra- and extra-cranial vessels in dogs have been imaged using conventional X-ray or digital subtraction angiography. As both of these methods are surgically demanding and involve the use of complex equipment with potential risks for the animal, they are now mainly used in experimental work. In this study, a method for delineating the intra- and extra-cranial vessels of dogs using contrast-enhanced magnetic resonance angiography (CE-MRA) has been examined. The MR images were acquired on a 1.5-T scanner after a bolus injection of gadolinium contrast medium. CE-MRA showed most of the major intra- and extra-cranial arteries and veins as well as most venous sinuses and plexuses of the brain. Difficulties may arise in differentiating between minor arterial and venous vessels as well as in the measurement of vessel diameter. More frequent use of this diagnostic method in clinical cases will indicate whether MRA of the head and brain is of as much use in veterinary medicine as it has become in human medicine. There is certainly potential value in detecting rare conditions in animals such as aneurysms and vascular malformations, as well as in examining vessels feeding intracranial masses and the investigation of cerebrovascular accidents.


Genotoxicity evaluation of kaurenoic acid, a bioactive diterpenoid present in Copaiba oil.
Copaiba oil extracted from the Amazon traditional medicinal plant Copaifera langsdorffii is rich in kaurenoic acid (ent-kaur-16-en-19-oic acid), a diterpene that has been shown to exert anti-inflammatory, hypotensive, and diuretic effects in vivo and antimicrobial, smooth muscle relaxant and cytotoxic actions in vitro. This study evaluated its potential genotoxicity against Chinese hamster lung fibroblast (V79) cells in vitro, using the Comet and the micronucleus assays. Kaurenoic acid was tested at concentrations of 2.5, 5,10, 30 and 60 microg/mL. The positive control was the methylmethanesulfonate (MMS). The duration of the treatment of V79 cells with these agents was 3h. The results showed that unlike MMS, kaurenoic acid (2.5, 5, and 10 microg/mL) failed to induce significantly elevated cell DNA damage or the micronucleus frequencies in the studied tests. However, exposure of V79 cells to higher concentrations of kaurenoic acid (30 and 60 microg/mL) caused significant increases in cell damage index and frequency. The data obtained provide support to the view that the diterpene kaurenoic acid induces genotoxicity.


Neuroprotective effect of paeoniflorin in the mouse model of Parkinson's disease through α-synuclein/protein kinase C δ subtype signaling pathway.
Paeoniflorin, an active component of Radix Paeoniae Alba, has a neuroprotective effect in Parkinson's animal models. However, its mechanism of action remains to be determined. In this study, we hypothesized that the neuroprotective effect of paeoniflorin occurs through the α-synuclein/protein kinase C δ subtype (PKC-δ) signaling pathway. We tested our hypothesis in the 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP)-induced mouse model of Parkinson's disease. We evaluated the effects of paeoniflorin on the expression levels of signal components of the α-synuclein/PKC-δ pathway, cellular apoptosis and motor performance. Our results demonstrated that paeoniflorin restored the motor performance impairment caused by MPTP, inhibited apoptosis, and protected the ultrastructure of neurons. Paeoniflorin treatment also resulted in the dose-dependent upregulation of an antiapoptotic protein, B-cell lymphoma-2, at the mRNA and protein levels, similar to the effects of the positive control, selegiline. In contrast, paeoniflorin treatment downregulated the expression of pro-apoptotic proteins BCL2-Associated X2, α-synuclein, and PKC-δ at the mRNA and protein levels, as well as the level of the activated form of nuclear factor kappa B (p-NF-κB p65). Thus, our results showed that paeoniflorin exerts its neuroprotective effect by regulating the α-synuclein/PKC-δ signaling pathway to reduce neuronal apoptosis.


Effect of Swell1 on regulating chondrocyte hypertrophy during the condylar osteochondral development process in mice.
Chondrocyte hypertrophy is a significant factor in cartilage development, yet the molecular mechanism for cell volume expand during the process is remains unclear. In the present study, the relationship between Swell1, a cell volume regulated anion channel, and chondrocyte hypertrophy was explored. The results reveal that the spatiotemporal expression of Swell1 was similar with the development process of hypertrophic chondrocytes in condyles. Through Col10a1 mediated knock out of Swell1 in hypertrophy chondrocytes, we found that there are less obvious boundary between different condylar cartilage layers in which increased hypertrophic chondrocytes were scattered in all three cartilage layers. The cortical bone mass and bone mineral density in the subchondral bone significantly increased. Additionally, knock out of Swell1 could increase the expression of OCN in the femur condyle. Based on the aforementioned findings, a conclusion could be drawn that Swell1 is a significant factor in chondrocyte hypertrophy during the condylar osteochondral development process, and there was some difference between the mandibular and femur condyles, which will provide some new clues for understanding the development of cartilage and related diseases.


Effects of simulation on emergency vehicle drivers' competency in training.
The number of accidents involving emergency vehicles is a major concern for emergency service providers. This study assessed the effectiveness of adding a driving simulator to a traditional training program. The sample consisted of students attending the National Emergency Medical Services Academy in Lafayette, Louisiana. Each participant self-scheduled two days in which he or she would attend the driving portion of the training class. A total of 52 participants were in the control group and 50 participants were in the treatment group. In addition to the classroom training, the treatment group used a driving simulator prior to driving on the competency course. The treatment group took significantly less time to drive through the competency course on the first run than did the control group. It also acquired significantly fewer penalty points on the first run and required significantly fewer runs to complete the course. Simulators can be effective training tools for teaching emergency vehicle drivers.


Change perception using visual transients: object substitution and deletion.
In three experiments we studied change detection and identification when no extraneous transients were present in the image at the time of change. Each image consisted of 12 different objects, sorted by color into three different levels of probability of change. In Experiment 1, change of one object was detected and identified frequently in objects having the highest probability of change (central interest), which we hypothesize were mainly visited by attention. Changes in other objects with a lower probability of change (marginal interest), however, although detected efficiently were unlikely to be identified. Identification improved for less attended objects if the changed stimulus simply disappeared, allowing visual persistence to hold information about the object until attention could be shifted to it (Experiment 2). Contrary to previous findings showing that response times (RTs) for luminance change detection in a multi-element display are not altered by attention, we found changes in objects of central interest to be detected faster than in objects of marginal interest when objects' identity was to be held in working memory. However, no differences in RTs emerged in the same change detection task when objects' identity was not stored in working memory (Experiment 3).


Characteristics of the Novel Potassium-Competitive Acid Blocker Vonoprazan Fumarate (TAK-438).
Proton pump inhibitors (PPIs) are widely prescribed as first-line therapy for the treatment of acid-related diseases, such as peptic ulcers and gastro-esophageal reflux disease, and for the eradication of Helicobacter pylori. However, the therapeutic efficacy of conventional PPIs is considered limited because: (1) they are unstable under acidic conditions and require an enteric-coated formulation in clinical use; (2) they show high interindividual variability in pharmacokinetics due to genetic polymorphisms of cytochrome P450 (CYP) 2C19 metabolism; (3) they have a relatively slow onset of pharmacological action and may require several doses to achieve optimal acid suppression and symptom relief; and (4) they often do not provide stable suppression of gastric acid secretion over 24 h. Vonoprazan fumarate (TAK-438, hereinafter referred to as "vonoprazan") is a new potassium-competitive acid blocker (P-CAB) developed to resolve the above limitations of conventional PPIs. Various physicochemical data have shown that vonoprazan has a high solubility and stability over a broad pH range in aqueous conditions. In addition, vonoprazan has a more potent and longer-lasting acid suppression effect than the conventional PPI, lansoprazole. Preclinical pharmacokinetic studies have shown that vonoprazan is accumulated and retained in the stomach for more than 24 h, even after it is eliminated from the plasma. From these findings, we propose that vonoprazan, which possesses a novel mode of action, can improve on the outcomes seen with conventional PPI-based treatments for acid-related diseases. This review project, including the publication of this article, was funded by Takeda Pharmaceutical Company Limited.


Moving On from the (Unimaginable) Year of the Nurse and the Midwife.
What can we take away from this year?


Mitral annuloplasty with the interatrial groove-left atrial dome approach in a patient with Marfan syndrome.
The choice of mitral valve surgical approach has always been a difficult problem in patients with small left atrium. We report a case of a patient with Marfan syndrome who underwent the David operation and mitral annuloplasty. The patient had a small left atrium, so we severed the superior vena cava and opened the interatrial groove and left atrial dome. This method allows for excellent exposure of the mitral valve and subvalvular apparatus, enabling a successful operation. The interatrial groove-left atrial dome approach provides an option for patients with a small left atrium undergoing mitral valve surgery.