A Randomized Controlled Single-Blind Study Demonstrating Superiority of Amniotic Suspension Allograft Injection Over Hyaluronic Acid and Saline Control for Modification of Knee Osteoarthritis Symptoms.
Placental-derived tissues are a known source of anti-inflammatory and immune modulating factors. Published pilot data on amniotic suspension allograft (ASA) for the treatment of osteoarthritis (OA) demonstrated safety and trends for improved pain and function. A multicenter randomized controlled trial was designed to evaluate the efficacy of symptom modulation with ASA compared with saline and hyaluronic acid (HA) in subjects with knee OA. A total of 200 subjects were randomized 1:1:1 to ASA, HA, or saline, with subjects blinded to their allocation. Changes from baseline of patient-reported outcomes (PROs)-EQ-5D-5L, Knee Osteoarthritis Outcome Score (KOOS), visual analog scale (VAS), Tegner, and Single Assessment Numerical Evaluation (SANE)-were compared between groups. Patients reporting unacceptable pain at 3 months were considered treatment failures and withdrawn from the study. Statistical analysis was completed by comparing changes in PROs from baseline to 3 and 6 months for all groups. Comparison of demographics between treatment groups showed no significant differences between groups. Patients reporting unacceptable pain at 3 months in each group were ASA (13.2%), HA (68.8%), and saline (75%). Patients receiving ASA demonstrated significantly greater improvements from baseline for overall pain (VAS), KOOS pain, and KOOS-activities of daily living scores compared with those in the HA group (3 months) and both groups (6 months). ASA patients had significantly greater improvements in KOOS symptom scores compared with HA and saline at 3 and 6 months, respectively. OMERACT-OARSI responder rates for ASA, HA, and saline groups were 69.1, 39.1, and 42.6%, respectively (p = 0.0007). Subjects receiving ASA treatment showed greater improvements in PROs and fewer patients reported unacceptable pain compared with HA and saline. The evidence presented in this Level I Randomized Controlled Trial suggests that ASA injection is an effective treatment for the nonoperative management of symptomatic knee OA.


Methods for Evaluating Emotions Evoked by Food Experiences: A Literature Review.
Besides sensory characteristics of food, food-evoked emotion is a crucial factor in predicting consumer's food preference and therefore in developing new products. Many measures have been developed to assess food-evoked emotions. The aim of this literature review is (i) to give an exhaustive overview of measures used in current research and (ii) to categorize these methods along measurement level (physiological, behavioral, and cognitive) and emotional processing level (unconscious sensory, perceptual/early cognitive, and conscious/decision making) level. This 3 × 3 categorization may help researchers to compile a set of complementary measures ("toolbox") for their studies. We included 101 peer-reviewed articles that evaluate consumer's emotions and were published between 1997 and 2016, providing us with 59 different measures. More than 60% of these measures are based on self-reported, subjective ratings and questionnaires (cognitive measurement level) and assess the conscious/decision-making level of emotional processing. This multitude of measures and their overrepresentation in a single category hinders the comparison of results across studies and building a complete multi-faceted picture of food-evoked emotions. We recommend (1) to use widely applied, validated measures only, (2) to refrain from using (highly correlated) measures from the same category but use measures from different categories instead, preferably covering all three emotional processing levels, and (3) to acquire and share simultaneously collected physiological, behavioral, and cognitive datasets to improve the predictive power of food choice and other models.


Intravenous digital subtraction angiography of arteriosclerotic vertebrobasilar disease.
Intravenous digital subtraction angiography (DSA) was performed in 111 patients with vertebrobasilar ischemia. Ninety percent of the vertebral images were of diagnostic quality; 23% of the basilar images were good quality and 53% fair quality; and 58% of the posterior cerebral images were poor. Compared with selective film arteriography in 23 patients, DSA tended to underestimate the degree of atheromatous disease. Segments of the basilar artery were often poorly seen, which could result in false-negative errors. DSA can provide a general assessment of atheromatous disease of the brachiocephalic vessels, including the vertebral and carotid arteries, and in many cases can exclude occlusion or critical stenosis of the vertebrobasilar system. However, it does not adequately image the posterior cerebral or cerebellar artery.


[Expression of malignancy traits in the interspecific somatic hybrids of tumor and normal cells].
Tumorigenicity and anchorage independence in two types of the interspecies hybrids of the tumor and normal mammalian cells were studied. One hybrid type was derived from fusion of spontaneously transformed Chinese hamster and normal mouse cells; the second type was obtained by fusion of SV40-transformed Djungarian hamster and the same mouse cells. The tumorigenicity in the athymic nude mice was suppressed in the first type of hybrids. The hybrid clones derived from fusion of SV40-transformed and normal cells could form tumor in nude mice. Testing of hybrid clones for their ability to form colonies in soft agar showed that all hybrids grew well in the medium, similar to tumor parental cells. These data suggest that malignancy and anchorage independence are under separate genetic control. The influence of the origin of the tumor parental cells (spontaneous or SV40-virus transformation) on the expression of the malignancy in hybrids of the tumor and normal cells is discussed.


Active elasticity drives the formation of periodic beading in damaged axons.
In several pathological conditions, such as coronavirus infections, multiple sclerosis, Alzheimer's and Parkinson's diseases, the physiological shape of axons is altered and a periodic sequence of bulges appears. Experimental evidences suggest that such morphological changes are caused by the disruption of the microtubules composing the cytoskeleton of the axon. In this paper, we develop a mathematical model of damaged axons based on the theory of continuum mechanics and nonlinear elasticity. The axon is described as a cylinder composed of an inner passive part, called axoplasm, and an outer active cortex, composed mainly of F-actin and able to contract thanks to myosin-II motors. Through a linear stability analysis we show that, as the shear modulus of the axoplasm diminishes due to the disruption of the cytoskeleton, the active contraction of the cortex makes the cylindrical configuration unstable to axisymmetric perturbations, leading to a beading pattern. Finally, the nonlinear evolution of the bifurcated branches is investigated through finite element simulations.


Risk factors and outcomes of IgA nephropathy recurrence after kidney transplantation: a systematic review and meta-analysis.
IgA nephropathy may recur in patients receiving kidney transplantation due to IgA nephropathy induced renal failure. The risk factors for recurrence are still at issue. The aim of this study was to conduct a systematic review and meta-analysis to assess risk factors and outcomes for IgA nephropathy recurrence. We used PubMed, EMBASE, Cochrane Library, Web of Science, Scopus, CNKI, WanFang, VIP and CBM to search for relevant studies published in English and Chinese. Cohort or case-control studies reporting risk factors or outcomes for IgA nephropathy recurrence were included. Fifty-eight studies were included. Compare to no recurrence group, those with IgAN recurrence had younger age (mean difference [MD]=-4.27 years; risk ratio [RR]=0.96), younger donor age (MD=-2.19 years), shorter time from IgA nephropathy diagnosis to end stage renal disease (MD=-1.84 years; RR=0.94), shorter time on dialysis (MD=-3.14 months), lower human leukocyte-antigen (HLA) mismatches (MD=-0.11) and HLA-DR mismatches (MD=-0.13). HLA-B46 antigen (RR=0.39), anti-IL-2-R antibodies induction (RR=0.68), mycophenolate mofetil (RR=0.69), and pretransplant tonsillectomy (RR=0.43) were associated with less IgAN recurrence. Of note, male recipient gender (RR=1.17), related donor (RR=1.53), retransplantation (RR=1.43), hemodialysis (RR=1.68), no induction therapy (RR=1.73), mTOR inhibitor (RR=1.51), angiotensin-converting enzyme inhibitors or angiotensin-receptor blockers (RR=1.63) were risk factors for IgAN recurrence. Recurrence increased the risk of graft loss (RR=2.19). This study summarized the risk factors for recurrence of IgA nephropathy after kidney transplantation. Well-designed prospective studies are warranted for validation. https://www.crd.york.ac.uk/PROSPERO/display_record.php?RecordID=377480, identifier CRD42022377480.


Does Fluoroscopy Induce DNA Oxidative Damage in Patients Undergoing Catheter Ablation?
As no studies before now have thoroughly investigated the risk associated with the exposure to low-dose ionizing radiations in patients undergoing catheter ablation (CA), we aimed to evaluate the oxidative and DNA damage in 59 CA patients (10 of whom received N-acetylcysteine (NAC) before the procedure). A burst of oxidized/reduced glutathione ratio was observed 3 hours after procedure that was diminished by NAC administration. 8-Hydroxy-2'-deoxyguanosine (8-OHdG) concentrations, index of oxidative DNA damage, showed a peak 24 hours after CA. A positive correlation between 8-OHdG peak and fluoroscopy time and a negative correlation among 8-OHdG decrease (from the peak to 48 hours after CA) and all procedure parameters were found. Furthermore, DNA tail percentages (which reflect the extent and the number of DNA strand breaks) positively correlated with 8-OHdG concentrations. This study evaluates for the first time the kinetic of oxidative damage in patients undergoing CA procedure. Our findings raise the question of whether 8-OHdG can be used as a circulating biomarker of DNA oxidative damage induced by low-dose ionizing radiations in different clinical settings. Antioxid. Redox Signal. 28, 1137-1143.


The protective effect of low-dose methotrexate on ischemia-reperfusion injury of the rabbit spinal cord.
Methotrexate was developed as a cytostatic agent, but at low doses, it has shown potent anti-inflammatory activity. Previous studies have demonstrated that the anti-inflammatory effects of methotrexate are primarily mediated by the release of adenosine. In this study, we hypothesized that low-dose methotrexate has protective effects in spinal cord ischemia-reperfusion injury. Rabbits were randomized into the following four groups of eight animals each: group 1 (control), group 2 (ischemia), group 3 (methylprednisolone) and group 4 (methotrexate). In the control group only a laparotomy was performed. In all the other groups, the spinal cord ischemia model was created by the occlusion of the aorta just caudal to the renal artery. Neurological evaluation was performed with the Tarlov scoring system. Levels of myeloperoxidase, malondialdehyde and catalase were analyzed, as were the activities of xanthine oxidase and caspase-3. Histopathological and ultrastructural evaluations were also performed. After ischemia-reperfusion injury, increases were found in the serum and tissue myeloperoxidase levels, tissue malondialdehyde levels, xanthine oxidase activity and caspase-3 activity. In contrast, both serum and tissue catalase levels were decreased. After the administration of a low-dose of methotrexate, decreases were observed in the serum and tissue myeloperoxidase levels, tissue malondialdehyde levels, xanthine oxidase activity and caspase-3 activity. In contrast, both the serum and tissue catalase levels were increased. Furthermore, low-dose methotrexate treatment showed improved results concerning the histopathological scores, the ultrastructural score and the Tarlov scores. Our results revealed that low-dose methotrexate exhibits meaningful neuroprotective activity following ischemia-reperfusion injury of the spinal cord.


Ileal resection: effect of cimetidine and taurine on intrajejunal bile acid precipitation and lipid solubilisation.
We have investigated whether pH-dependent bile acid precipitation limits lipid solubilisation after ileal resection, and whether treatment with cimetidine, or taurine improves solubilisation. Nine ileal resection patients were treated with placebo, cimetidine and taurine in random order for two weeks each. Upper jejunal content was aspirated and pooled according to pH for three hours after a standard Lundh test meal. On placebo, 50% of the bile acids were precipitated at pH less than 5, compared with only 26% at pH greater than 6, whilst aqueous-phase lipid concentration tended to be lower at pH less than 5 than at pH greater than 6 (5.1 vs 8.2 mmol/l). On cimetidine mean pH rose, particularly during the third hour (6.6 vs 5.8, p less than 0.05), associated with a reduction in bile acid precipitation (13.9 vs 33.1%, p less than 0.05), and an increase in aqueous-phase lipid concentration (10.4 vs 6.6 mmol/l, p less than 0.05). On taurine, the proportion of taurine conjugated bile acids increased (67 vs 22%, p less than 0.01), but there was no significant change in bile acid precipitation or lipid solubilisation. Lower jejunal samples were aspirated similarly from five of these patients on no treatment, and all were at pH greater than 6; apparent 'precipitation' was reduced (16.4 vs 28.1%), but lipid solubilisation did not improve. These findings suggest that pH dependent bile acid precipitation can limit lipid solubilisation within the jejunum after ileal resection, and that these effects can be reduced by cimetidine but not by taurine. Cimetidine may have a role in ileal resection patients with severe steatorrhoea unresponsive to dietary fat restriction.


Procedural outcomes associated with transvenous lead extraction in patients with abandoned leads: an ESC-EHRA ELECTRa (European Lead Extraction ConTRolled) Registry Sub-Analysis.
The decision to abandon or extract superfluous leads remains controversial. We sought to compare procedural outcome of patients with and without abandoned leads undergoing transvenous lead extraction (TLE). An analysis of the ESC-EHRA European Lead Extraction ConTRolled ELECTRa registry was conducted. Patients were stratified into two groups based on the presence (Group 1) or absence (Group 2) of abandoned leads at the time for extraction. Out of 3508 TLE procedures, 422 patients (12.0%) had abandoned leads (Group 1). Group 1 patients were older and more likely to have implantable cardioverter-defibrillator devices, infection indication (78.8% vs. 49.8%), and vegetations (24.6% vs. 15.3%). Oldest lead dwelling time was longer in Group 1 (10.9 vs. 6.3 years) as was the number of extracted leads per patient (3.2 vs. 1.7). Manual traction failure (94.5% vs. 78.8%), powered sheath use (50.7% vs. 28.4%), and femoral approach were higher in Group 1 (P < 0.0001). Procedural success rate and clinical success (89.8% vs. 96.6%, P < 0.0001) were lower in Group 1. Major complication including deaths (5.5% vs. 2.3%, P = 0.0007) and procedure related major complications (3.3% vs. 1.4%, P = 0.0123) were higher in Group 1. The presence of abandoned leads at the time of TLE was an independent predictor of clinical failure [odds ratio (OR) 2.31, confidence interval (CI) 1.57-3.40] and complications [OR 1.69, CI 1.22-2.35]. receiver-operating characteristic curve analysis showed a dwell time threshold of 9 years for radiological failure and major complications. Previously abandoned leads at the time of TLE were associated with increased procedural complexity, clinical failure, and major complication, which may have important implications for future studies regarding managing of lead failures.


[para-Aminobenzoic acid as a sole source of carbon and energy for Pseudomonas desmoliticum].
A strain of Psuedomonas desmoliticum has been isolated from soil. It utilizes, as a source of carbon and energy, p-aminobenozic (PABA), p-fluorobenzoic acids, and some other aromatic compounds. The strain has been isolated by inoculating soil suspensions onto Petri plates with a solid mineral medium containing 0.1% PABA as a carbon source. The preparatory metabolism of PABA was studied in this work; p-hydroxybenzoic and protocatechuic acids were found to be its intermediate products. Enzyme systems catalysing oxidation of aromatic compounds and glucose are inducible.


Towards a database for genotype-phenotype association research: mining data from encyclopaedia.
To associate phenotypic characteristics of an organism to molecules encoded by its genome, there is a need for well-structured genotype and phenotype data. We use a novel method for extracting data on phenotype and genotype characteristics of microorganisms from text. As a resource, we use an encyclopedia of microorganisms, which holds phenotypic and genotypic data and create a structured, flexible data resource, which can be exported to a range of database formats, containing genotype and phenotype data for 2412 species and 873 genera of microbes. This data source has great potential as a resource for future biological research on genotype-phenotype associations. In this paper, we focus on describing the structure and content of the resulting database and on evaluating the method used for extracting the data. We conclude that the resulting database can be used as a reliable complementary resource for research into genotype-phenotype association.


The probiotic supplementation role in improving the immune system among people with ulcerative colitis: a narrative review.
The purpose of this paper is to summarize the current evidence on probiotics' uses as an adjuvant for ulcerative colitis (UC) and provide an understanding of the effect of probiotics supplement on the immune system and inflammatory responses among UC patients and subsequent therapeutic benefits. A narrative review of all the relevant published papers known to the author was conducted. UC is a chronic inflammatory bowel disease (IBD) that results in inflammation and ulceration of the colon and rectum. The primary symptoms of active disease are diarrhea, abdominal pain, and rectal bleeding. About 70% of the human immune system (mucosal-associated lymphoid tissue) originates in the intestine. Probiotics are live microorganisms that help in stabilizing the gut microbiota (nonimmunologic gut defense), restores normal flora, and enhance the humoral immune system. Probiotics especially Bifidobacterium, Saccharomyces boulardii, and lactic acid-producing bacteria have been used as an adjunct therapy for treating UC to ameliorate disease-related symptoms and reduce relapse rate. Probiotics, in general, modulate the immune system through their ability to enhance the mucosal barrier function, or through their interaction with the local immune system to enhance regulatory T cell responses, decrease the pro-inflammatory cytokines such as tumor necrosis factor alpha and interleukin 1 beta and increase anti-inflammatory factor interleukin 10. More studies are needed to explore the properties of the various probiotic bacterial strains, their different uses, as well as the dosage of probiotics and duration for treating different disorders. Further clinical investigations on mechanisms of action and how probiotics modulate the immune system may lead to further advances in managing IBD.


Carbon ion beam is more effective to induce cell death in sphere-type A172 human glioblastoma cells compared with X-rays.
To obtain human glioblastoma cells A172 expressing stem cell-related protein and comparison of radiosensitivity in these cells with X-rays and carbon beam. Human monolayer-type A172 glioblastoma cells were maintained in normal medium with 10% bovine serum. In order to obtain sphere-type A172 cells the medium was replaced with serum-free medium supplemented with growth factors. Both types of A172 cells were irradiated with either X-rays or carbon ion beams and their radiosensitivity was evaluated. Serum-free medium induced expression of stem cell-related proteins in A172 cells along with the neurosphere-like appearance. These sphere-type cells were found resistant to both X-rays and carbon ion beams. Phosphorylation of histone H2A family member X persisted for a longer period in the cells exposed to carbon ion beams than in those exposed to X-rays and it disappeared quicker in the sphere type than in the monolayer type. Relative radioresistance of the sphere type cells was smaller for carbon ion beams than for X-rays. We demonstrated that glioblastoma A172 cells with induced stem cell-related proteins turned resistant to irradiation. Accelerated heavy ion particles may have advantage over X-rays in overcoming the tumor resistance due to cell stemness.


Cell system ontology: representation for modeling, visualizing, and simulating biological pathways.
With the rapidly accumulating knowledge of biological entities and networks, there is a growing need for a general framework to understand this information at a system level. In order to understand life as system, a formal description of system dynamics with semantic validation will be necessary. Within the context of biological pathways, several formats have been proposed, e.g., SBML, CellML, and BioPAX. Unfortunately, these formats lack the formal definitions of each term or fail to capture the system dynamics behavior. Thus, we have developed a new system dynamics centered ontology called Cell System Ontology (CSO). As an exchange format, the ontology is implemented in the Web Ontology Language (OWL), which enables semantic validation and automatic reasoning to check the consistency of biological pathway models. The features of CSO are as follows: (1) manipulation of different levels of granularity and abstraction of pathways, e.g., metabolic pathways, regulatory pathways, signal transduction pathways, and cell-cell interactions; (2) capture of both quantitative and qualitative aspects of biological function by using hybrid functional Petri net with extension (HFPNe); and (3) encoding of biological pathway data related to visualization and simulation, as well as modeling. The new ontology also predefines mature core vocabulary, which will be necessary for creating models with system dynamics. In addition, each of the core terms has at least one standard icon for easy modeling and accelerating the exchangeability among applications. In order to demonstrate the potential of CSO-based pathway modeling, visualization, and simulation, we present an HFPNe model for the ASEL and ASER regulatory networks in Caenorhabditis elegans.


New-onset hyperglycemia immediately after liver transplantation: A national survey from China Liver Transplant Registry.
New-onset hyperglycemia (NOH) is a common phenomenon after liver transplantation (LT), but its impact on clinical outcomes has not yet been fully assessed. We aimed to evaluate the etiology and prognosis of NOH within 1 month after LT. The data of 3339 adult patients who underwent primary LT from donation after citizen death between January 2010 and June 2016 were extracted from China Liver Transplant Registry database and analyzed. NOH was defined as fasting blood glucose ≥7.0 mmol/L confirmed on at least two occasions within the first post-transplant month with or without hypoglycemic agent. Of 3339 liver recipients, 1416 (42.4%) developed NOH. Recipients with NOH had higher incidence of post-transplant complications such as graft and kidney failure, infection, biliary stricture, cholangitis, and tumor recurrence in a glucose concentration-dependent manner as compared to non-NOH recipients (P < 0.05). The independent risk factors of NOH were donor warm ischemic time >10 min, cold ischemic time >10 h, anhepatic time >60 min, recipient model for end-stage liver disease score >30, moderate ascites and corticosteroid usage (P < 0.05). Liver enzymes (alanine aminotransferase and gamma-glutamyltranspeptidase) on post-transplant day 7 significantly correlated with NOH (P < 0.001). NOH leads to increased morbidity and mortality in liver recipients. Close surveillance and tight control of blood glucose are desiderated immediately following LT particularly in those with delayed graft function and receiving corticosteroid. Strategic targeting graft ischemic injury may help maintain glucose homeostasis.


Polybenzofulvene derivatives bearing dynamic binding sites as potential anticancer drug delivery systems.
In order to obtain new advanced functional materials capable of recognizing drug molecules, the polybenzofulvene backbone of molecular brush poly-6-MOEG-9-TM-BF3k has been functionalized with a "synthetic dynamic receptor" composed of two 1-adamantylurea moieties linked together by means of a dipropyleneamino bridge as in Meijer's bis(adamantylurea) pincer (BAUP). This functional material, bearing synthetic receptors potentially capable of recognizing/loading and then delivering drug molecules, was used to prepare colloidal drug delivery systems (by means of soft interaction with BAUP) for delivering the model anti-cancer drug doxorubicin (DOXO). The resulting nanostructured drug delivery systems containing the physically loaded drug were characterized in terms of drug loading and release, dimensions and zeta potential, and in vitro cell activity and uptake on two different cell lines (i.e. the human bronchial epithelial 16HBE and the human colon cancer HCT116). On normal cells, free DOXO was found to be more cytotoxic than DOXO-loaded nanogels at the higher tested concentration and, only on cancer cells, DOXO-loaded nanogels show similar or slightly higher cytotoxicity values than free DOXO, suggesting potential advantages in the treatment of cancer. These results were supported by fluorescence microscopy studies, which suggested that DOXO-loaded nanogels provide an extracellular reservoir of the drug, which is gradually released and internalized within the cells.


Femoral mechanics in the lesser bushbaby (Galago senegalensis): structural adaptations to leaping in primates.
One method used to examine the relationship between behavioral strategies and anatomical adaptation is to study the results of mechanical stress associated with a given behavior and compare this with skeletal adaptations to other behaviors. This comparative approach is appropriate for highlighting combinations of features that are specializations to specific types of behavior. The purpose of this paper is to compare femoral mechanics in Galago senegalensis with previously collected data for macaques and humans as a basis for discussing structural adaptations in the primate hindlimb to leaping. The stiffness and load carrying capabilities of the femoral diaphyses of 27 G. senegalensis were analyzed using the SCADS computer program. The data suggest that the galago femur is well adapted to sustain large sagittal plane compressive loads rather than large bending loads. The straightness of the femoral shaft and large midshaft area moments of inertia prevent buckling from these large compressive loads. Calculations indicate that the ratio of critical buckling load to body weight in galago is 31 times that in macaques and 55 times that in humans. The femur of this saltatory primate is morphologically adapted to resist buckling when subjected to large compressive loads, while those of macaques and humans are better adapted to resist bending moments caused by ground reaction forces acting on the extended limb. The differences between galago on the one hand and macaques and humans on the other suggest that relatively smaller moments about the hip and relatively larger moments about the knee accompany more quadrupedal and bipedal walking, while habitual leaping is associated with relatively larger moments about the hip. These data reinforce the apparent similarity of the mechanical effects of quadrupedal and bipedal locomotion on the femur and dissimilarity with femoral mechanics in habitually saltatory primates.


Nucleotide sequence analysis and overexpression of the gene encoding a type III chloramphenicol acetyltransferase.
The gene catIII, encoding a type III enterobacterial chloramphenicol acetyltransferase, was cloned from the transmissible plasmid R387 into pBR322 and bacteriophage M13 mp8. Nucleotide sequence analysis of 1160 bp of DNA identified an open reading frame encoding a protein of 213 amino acid residues and a calculated molecular mass of 24965 Da. The predicted N-terminal sequence is identical with that determined by Edman degradation of chloramphenicol acetyltransferase purified from Escherichia coli harbouring R387. Sequences equivalent to the consensus motifs for initiation and rho-factor-independent termination of transcription in E. coli occur 5' and 3' to the catIII open reading frame. In contrast with the catI gene, present on transposon Tn9 and many enterobacterial plasmids, expression of catIII is not subject to cyclic AMP-mediated catabolite repression in vivo and there is no sequence in the 5' non-coding DNA that resembles that deduced as the consensus for the binding of cyclic AMP receptor protein. Unique restriction-endonuclease cleavage sites were introduced adjacent to the catIII reading frame by using oligonucleotide-directed mutagenesis to facilitate insertion into E. coli expression vectors. Fully active chloramphenicol acetyltransferase represents 30-50% of the soluble protein component of cell-free extracts of E. coli containing the appropriate plasmids.


Exploring the Molecular Interactions of Symmetrical and Unsymmetrical Selenoglycosides with Human Galectin-1 and Galectin-3.
Galectins (Gals) are small cytosolic proteins that bind β-galactoside residues via their evolutionarily conserved carbohydrate recognition domain. Their dysregulation has been shown to be associated with many diseases. Consequently, targeting galectins for clinical applications has become increasingly relevant to develop tailored inhibitors selectively for one galectin. Accordingly, binding studies providing the molecular details of the interaction between galectin and inhibitor may be useful for the rational design of potent and selective antagonists. Gal-1 and Gal-3 are among the best-studied galectins, mainly for their roles in cancer progression; therefore, the molecular details of their interaction with inhibitors are demanded. This work gains more value by focusing on the interaction between Gal-1 and Gal-3 with the selenylated analogue of the Gal inhibitor thiodigalactose, characterized by a selenoglycoside bond (SeDG), and with unsymmetrical diglycosyl selenides (unsym(Se). Gal-1 and Gal-3 were produced heterologously and biophysically characterized. Interaction studies were performed by ITC, NMR spectroscopy, and MD simulation, and thermodynamic values were discussed and integrated with spectroscopic and computational results. The 3D complexes involving SeDG when interacting with Gal-1 and Gal-3 were depicted. Overall, the collected results will help identify hot spots for the design of new, better performing, and more specific Gal inhibitors.


Asexual Female Gametogenesis Involves Contact with a Sexually-Fated Megaspore in Apomictic Hieracium.
Apomixis results in asexual seed formation where progeny are identical to the maternal plant. In ovules of apomictic species of the Hieracium subgenus Pilosella, meiosis of the megaspore mother cell generates four megaspores. Aposporous initial (AI) cells form during meiosis in most ovules. The sexual pathway terminates during functional megaspore (FM) differentiation, when an enlarged AI undergoes mitosis to form an aposporous female gametophyte. Then, the mitotically programmed FM dies along with the three other megaspores by unknown mechanisms. Transcriptomes of laser-dissected AIs, ovule cells, and ovaries from apomicts and AI-deficient mutants were analyzed to understand the pathways involved. The steps leading to AI mitosis and sexual pathway termination were determined using antibodies against arabinogalactan protein epitopes found to mark both sexual and aposporous female gametophyte lineages at inception. At most, four AIs differentiated near developing megaspores. The first expanding AI cell to contact the FM formed a functional AI that underwent mitosis soon after megaspore degeneration. Transcriptome analyses indicated that the enlarged, laser-captured AIs were arrested in the S/G2 phase of the cell cycle and were metabolically active. Further comparisons with AI-deficient mutants showed that AIs were enriched in transcripts encoding homologs of genes involved in, and potentially antagonistic to, known FM specification pathways. We propose that AI and FM cell contact provides cues required for AI mitosis and megaspore degeneration. Specific candidates to further interrogate AI-FM interactions were identified here and include Hieracium arabinogalactan protein family genes.


Theory of molecular excitation and relaxation near a plasmonic device.
The new optical concepts currently developed in the research field of plasmonics can have significant practical applications for integrated optical device miniaturization as well as for molecular sensing applications. Particularly, these new devices can offer interesting opportunities for optical addressing of quantum systems. In this article, we develop a realistic model able to explore the various functionalities of a plasmon device connected to a single fluorescing molecule. We show that this theoretical method provides a useful framework to understand how quantum and plasmonic entities interact in a small area. Thus, the fluorescence signal evolution from excitation control to relaxation control depending on the incident light power is clearly observed.


Peak Fat Oxidation Rate Is Closely Associated With Plasma Free Fatty Acid Concentrations in Women; Similar to Men.
Introduction: In men, whole body peak fat oxidation (PFO) determined by a graded exercise test is closely tied to plasma free fatty acid (FFA) availability. Men and women exhibit divergent metabolic responses to fasting and exercise, and it remains unknown how the combined fasting and exercise affect substrate utilization in women. We aimed to investigate this, hypothesizing that increased plasma FFA concentrations in women caused by fasting and repeated exercise will increase PFO during exercise. Then, that PFO would be higher in women compared with men (data from a previous study). Methods: On two separate days, 11 young endurance-trained women were investigated, either after an overnight fast (Fast) or 3.5 h after a standardized meal (Fed). On each day, a validated graded exercise protocol (GXT), used to establish PFO by indirect calorimetry, was performed four times separated by 3.5 h of bed rest both in the fasted (Fast) or fed (Fed) state. Results: Peak fat oxidation increased in the fasted state from 11 ± 3 (after an overnight fast, Fast 1) to 16 ± 3 (mean ± SD) mg/min/kg lean body mass (LBM) (after ~22 h fast, Fast 4), and this was highly associated with plasma FFA concentrations, which increased from 404 ± 203 (Fast 1) to 865 ± 210 μmol/L (Fast 4). No increase in PFO was found during the fed condition with repeated exercise. Compared with trained men from a former identical study, we found no sex differences in relative PFO (mg/min/kg LBM) between men and women, in spite of significant differences in plasma FFA concentrations during exercise after fasting. Conclusion: Peak fat oxidation increased with fasting and repeated exercise in trained women, but the relative PFO was similar in young trained men and women, despite major differences in plasma lipid concentrations during graded exercise.


Safety and efficacy of a Bluetongue inactivated vaccine (serotypes 1 and 4) in sheep.
A new inactivated vaccine against Bluetongue virus (BTV) serotypes 1 and 4, was developed from field isolates. Safety and efficacy of the vaccine were evaluated in sheep by serological monitoring and virus nucleic acid detection after experimental infection of vaccinated animals. Seroconversion was observed in vaccinated animals at day 14 post vaccination (pv) with neutralizing antibody titer of 1.9 and 1.8 for serotypes 1 and 4, respectively. The titer increase significantly after the booster reaching 2.7 and persist one year >1.5 for both serotypes. After challenge with virulent isolates, vireamia was recorded in control animals, as evident by q-PCR with threshold cycles (Ct) ranging from 24 to 31 and peaked at day 10 post challenge, while no vireamia was detected in vaccinated animals. Vaccinated sheep were fully protected against the disease and infection.


Associations of adolescent hopelessness and self-worth with pregnancy attempts and pregnancy desire.
We examined the associations of pregnancy desire (ambivalence or happiness about a pregnancy in the next year) and recent pregnancy attempts with hopelessness and self-worth among low-income adolescents. To evaluate independent associations among the study variables, we conducted gender-stratified multivariable logistic regression analyses with data derived from 2285 sexually experienced 9- to 18-year-old participants in the Mobile Youth Survey between 2006 and 2009. Fifty-seven percent of youths reported a desire for pregnancy and 9% reported pregnancy attempts. In multivariable analyses, hopelessness was positively associated and self-worth was negatively associated with pregnancy attempts among both female and male youths. Hopelessness was weakly associated (P = .05) with pregnancy desire among female youths. The negative association of self-worth and the positive association of hopelessness with pregnancy attempts among young men as well as young women and the association of hopelessness with pregnancy desire among young women raise questions about why pregnancy is apparently valued by youths who rate their social and cognitive competence as low and who live in an environment with few options for material success.


Treatment gap and mental health service use among Syrian refugees in Sultanbeyli, Istanbul: a cross-sectional survey.
Syrian refugees may have increased mental health needs due to the frequent exposure to potentially traumatic events and violence experienced during the flight from their home country, breakdown of supportive social networks and daily life stressors related to refugee life. The aim of this study is to report evidence on mental health needs and access to mental health and psychosocial support (MHPSS) among Syrians refugees living in Sultanbeyli-Istanbul, Turkey. A cross-sectional survey was conducted among Syrian refugees aged 18 years or over in Sultanbeyli between February and May 2018. We used random sampling to select respondents by using the registration system of the municipality. Data among 1678 Syrian refugees were collected on mental health outcomes using the Posttraumatic Stress Disorder (PTSD) Checklist (PCL-5) and the Hopkins Symptoms Checklist (HSCL-25) for depression and anxiety. We also collected data on health care utilisation, barriers to seeking and continuing care as well as knowledge and attitudes towards mental health. Descriptive analyses were used. The estimated prevalence of symptoms of PTSD, depression and anxiety was 19.6, 34.7 and 36.1%, respectively. In total, 249 respondents (15%) screened positive for either PTSD, depression or anxiety in our survey and self-reported emotional/behavioural problems since arriving in Sultanbeyli. The treatment gap (the proportion of these 249 people who did not seek care) was 89% for PTSD, 90% for anxiety and 88% for depression. Several structural and attitudinal barriers for not seeking care were reported, including the cost of mental health care, the belief that time would improve symptoms, fear of being stigmatised and lack of knowledge on where and how to get help. Some negative attitudes towards people with mental health problems were reported by respondents. Syrian refugees hardly access MHPSS services despite high mental health needs, and despite formally having access to the public mental health system in Turkey. To overcome the treatment gap, MHPSS programmes need to be implemented in the community and need to overcome the barriers to seeking care which were identified in this study. Mental health awareness raising activities should be provided in the community alongside the delivery of psychological interventions. This is to increase help-seeking and to tackle negative attitudes towards mental health and people with mental health problems.


Steroid and growth factor modulation of aromatase activity in MCF7 and T47D breast carcinoma cell lines.
The effect of a number of steroids, growth factors, and peptides on aromatase activity in two estrogen receptor positive breast cancer cell lines (MCF7 and T47D) was investigated. The cells were incubated in Dulbecco's minimum essential medium containing phenol red and 10% fetal calf serum. Pronounced differences in basal aromatase activity and different responses to the addition of experimental agents were found in the two cell lines. Aromatase activity in MCF7 cells was significantly stimulated by phorbol 12,13-diacetate [PDA], dibutyryl cyclic AMP [(Bu)2cAMP], transforming growth factor alpha, and epidermal growth factor individually and PDA and (Bu)2cAMP in combination, while it was inhibited by dexamethasone and unaffected by transforming growth factor beta, fibroblast growth factor, platelet-derived growth factor, prolactin, and tamoxifen. Addition of cortisol to MCF7 cells had no effect on aromatase activity at 1 nM, caused suppression of activity at 10 nM and stimulated activity at 100 nM. Aromatase activity in T47D cells was stimulated by transforming growth factor alpha, epidermal growth factor, platelet-derived growth factor, prolactin, dexamethasone, and cortisol individually and PDA and (Bu)2cAMP in combination. It was unaffected by transforming growth factor beta, PDA, (Bu)2cAMP, and fibroblast growth factor. These findings suggest that aromatase activity is induced by agents which stimulate cyclic AMP-dependent protein kinases [e.g., (Bu)2cAMP] and that this effect is potentiated by factors which stimulate protein kinase C [e.g., PDA]. The effect on aromatase activity of growth factors, the actions of which are believed to be mediated by receptors linked to tyrosine kinase activity, is not as clearly defined, with a factor causing stimulation, inhibition, and no change in activity depending on the tissue concerned. Further insight into these differences will require resolution of the molecular mechanisms that mediate the actions of stimulatory and repressive growth factors on aromatase activity of oestrogen-producing cells.


Reimagining IRB review to incorporate a clear and convincing standard of evidence.
This commentary is a critical response to the article written by David Resnik regarding the use of a standard of evidence for Institutional Review Board (IRB) decision making. Resnik suggests that IRBs should not only base decisions on evidence, but that this evidence should be sufficient to ensure a "clear and convincing" standard similar to that used by juries for legal proceedings. We agree that the increased use of evidence to meet this standard would be ideal since this provides clear guidance and could allow for a more transparent IRB review. However, to effectively meet this standard, significant modification would be required for researchers as well as for IRBs' processes. First, researchers would be required to identify, understand and include appropriate scientific and ethics evidence in support of their protocol. IRB members and IRB professionals would need to discuss the importance, value, and significance of evidence in order to come to a collective decision regarding each protocol. Such responsibilities are justifiable and could bring much needed rigor and transparency to the system but they would require time, training, research, and education. While Resnik's suggestion seems to incorporate a small change with respect to a standard, in application it would actually require a novel system.


alpha,beta-epoxy vinyl triflates in Pd-catalyzed reactions.
[reaction: see text] Reactions of steroidal alpha,beta-epoxy vinyl triflates in Pd-catalyzed reactions are described. Oxidative insertion of Pd(0) into the C-O bond, giving vinylpalladium 12, is faster than formation of the pi-allyl derivative from the vinyl epoxide. Although 12 can be trapped under certain conditions, it eventually rearranges to palladium alkoxide 14, which is in equilibrium with 15 and/or 10.


The vomeronasal organ.
The vomeronasal organ (VNO) is a chemoreceptor organ enclosed in a cartilaginous capsule and separated from the main olfactory epithelium. The vomeronasal neurons have two distinct types of receptor that differ from each other and from the large family of odorant receptors. The VNO receptors are seven-transmembrane receptors coupled to GTP-binding protein, but appear to activate inositol 1,4,5-trisphosphate signaling as opposed to cyclic adenosine monophosphate. The nature of stimulus access suggests that the VNO responds to nonvolatile cues, leading to activation of the hypothalamus by way of the accessory olfactory bulb and amygdala. The areas of hypothalamus innervated regulate reproductive, defensive, and ingestive behavior as well as neuroendocrine secretion.


Phthalate pathway of phenanthrene metabolism: formation of 2'-carboxybenzalpyruvate.
The metabolism of phenanthrene by a gram-negative organism able to use this compound as a sole source of carbon and energy has been examined. 1-Hydroxy-2-naphthoic acid was oxidized by oxygen in a reaction catalyzed by a dioxygenase which was activated by ferrous ions. The stoichiometry of the oxidation and the UV spectrum of the product were consistent with the identification of the product as 2'-carboxybenzalpyruvate. This was confirmed by cleaving the product with a partially purified aldolase to yield 2-carboxybenzaldehyde and pyruvate. A number of enzymes for the metabolism of 1-hydroxy-2-naphthoic acid were induced by growth on phthalate or (less well) by growth on protocatechuate. The latter supported only a slow rate of growth, and this and poor induction may have been due to a slow rate of entry into the cell.


Connexin 46 (cx46) gap junctions provide a pathway for the delivery of glutathione to the lens nucleus.
Maintenance of adequate levels of glutathione (GSH) in the lens nucleus is critical for protection of lens proteins from the effects of oxidative stress and for lens transparency. How GSH is transported to the nucleus is unknown. We show that GSH diffuses to the nucleus from the outer cortex, where a high concentration of the anti-oxidant is established by synthesis or uptake, via the network of gap junctions. Using electrophysiological measurements, we found that channels formed by Cx46 and Cx50, the two connexin isoforms expressed in the lens, were moderately cation-selective (P(Na)/P(Cl) ∼5 for Cx46 and ∼3 for Cx50). Single channel permeation of the larger GSH anion was low but detectable (P(Na)/P(GSH) ∼12 for Cx46 and ∼8 for Cx50), whereas permeation of divalent anion glutathione disulfide (GSSG) was undetectable. Measurement of GSH levels in the lenses from connexin knock-out (KO) mice indicated Cx46, and not Cx50, is necessary for transport of GSH to the core. Levels of GSH in the nucleus were markedly reduced in Cx46 KO, whereas they were unaffected by Cx50 KO. We also show that GSH delivery to the nucleus is not dependent on the lens microcirculation, which is believed to be responsible for extracellular transport of other nutrients to membrane transporters in the core. These results indicate that glutathione diffuses from cortical fiber cells to the nucleus via gap junction channels formed by Cx46. We present a model of GSH diffusion from outer cells to inner fiber cells through gap junctions.


Benzyl p-guanidinothiobenzoate hydrochloride, a new active-site titrant for trypsin and trypsin-like enzymes.
Benzyl p-guanidinothiobenzoate hydrochloride was synthesized and demonstrated to be useful for active-site titration of bovine trypsin, bovine thrombin, human lung tryptase, bovine activated protein C, human Factor XIIa fragment and bovine Factor Xa beta. The titration is based on rapid formation of a stable acyl-enzyme with a stoichiometric release of benzyl thiol. Thiol production is measured quantitatively by including 4,4'-dithiodipyridine in the reaction mixture and measuring the increase in absorbance at 324 nm. Ellman's reagent has also been successfully employed, allowing measurement at 410 nm. Unlike p-nitrophenyl p'-guanidinobenzoate, the thioester titrant reacts slowly with chymotrypsin A alpha thus eliminating interference by this enzyme in most titrations. Advantages of this reagent as a titrant include: flexibility in detection of the released thiol, selectivity between trypsin and chymotrypsin-like enzymes, minimal pH-dependence of the epsilon of the absorbing species, relative stability of the reagent under titration conditions, and high epsilon at pH 7.2 with either 4,4'-dithiodipyridine or Ellman's reagent. The reagent should prove useful as an alternative to p-nitrophenyl p'-guanidinobenzoate hydrochloride for the determination of active-site concentrations of the enzymes employed, as well as of other related enzymes.


[Angiological complications following high tibial head correcting osteotomy--a case report].
Vascular complications following orthopedic procedures at the lower extremity are mainly thrombotic or thromboembolic events. Damages to vascular structures during operations close to the knee joint, like injuries of the popliteal artery, are extremely rare and receive a reserved consideration in the evaluation of postoperative complications. The extent of the vessel injury affects the grade of clinical symptoms-from acute arterial bleeding as far as slowly developing disturbances of haemodynamic, with effects to the different functions and pattern of the concerned tissues. Arterial bleeding into muscle compartments do not necessarily appear as an acute emergency, therefore it may occasionally be difficult to place the clinical symptoms into the right context of pathophysiological processes. In the present case a deep vein thrombosis of the affected extremity was initiated by a haemorrhage into the fossa popliteal and the compartments of the lower leg after injury of the popliteal artery. Subsequently a compartment syndrome, a false aneurysm of the popliteal artery and an arteriovenous fistula from popliteal artery into a lower leg vein developed.


Selective parenteral and enteral antisepsis regimen (SPEAR).
With infection rates approaching 80% for patients staying in ICU for more than 5 days there is clearly a need for effective prophylaxis. SPEAR is a system of antimicrobial prophylaxis which extends the regimen of selective digestive tract decontamination to encompass systemic antimicrobial prophylaxis and includes oral non-absorbable antibacterial and antifungal agents with initial use of a broad-spectrum systemic antibacterial.


Neurological paraneoplastic syndromes: an update.
To describe recent advances in the diagnosis and treatment of paraneoplastic neurological syndromes (PNS). PNS are rare complications of cancer caused by an immune cross-reaction between antigens expressed by tumor cells and neurons. The target of the immune attack can be an intracellular antigen or a cell-surface antigen. Although both types of autoimmunity are 'paraneoplastic', as indirectly triggered by the presence of a tumor, they profoundly differ in terms of clinical profile, pathogenesis and outcome. PNS associated with antibodies to intracellular antigens (icPNS) are characterized by relentless progression and poor response to treatment, because of rapid and permanent neuronal loss. PNS associated with antibodies to cell-surface antigens (csPNS) generally show favorable response to immune therapy and good functional outcome, as they result from reversible neuronal dysfunction. The spectrum of paraneoplastic autoimmunity has dramatically expanded following the discovery of cell-surface antibodies. Novel antibodies are incessantly discovered, some of which have a solid association with cancer. As csPNS usually respond to immune therapy, the optimization of current treatment strategies should have high priority to improve therapeutic results and prevent relapses.


Young coronary artery disease in patients undergoing percutaneous coronary intervention.
Understanding the nature and pattern of young coronary artery disease (CAD) is important due to the tremendous impact on these patients' socio-economic and physical aspect. Data on young CAD in the southeast Asian region is rather patchy and limited. Hence we utilized our National Cardiovascular Disease Database (NCVD)-Percutaneous Coronary Intervention (PCI) Registry to analyze young patients who underwent PCI in the year 2007 to 2009. This is a retrospective study of all patients who had undergone coronary angioplasty from 2007 to 2009 in 11 hospitals across Malaysia. Data were obtained from the NCVD-PCI Registry, 2007 to 2009. Patients were categorized into 2 groups-young and old, where young was defined as less than 45 years for men and less than 55 years for women and old was defined as more than or equals to 45 years for men and more than or equals to 55 years for women. Patients' baseline characteristics, risk factor profile, extent of coronary disease and outcome on dis.charge, and 30-day and 1-year follow-up were compared between the 2 groups. We analyzed 10268 patients, and the prevalence of young CAD was 16% (1595 patients). There was a significantly low prevalence of Chinese patients compared to other major ethnic groups. Active smoking (30.2% vs 17.7%) and obesity (20.9% vs 17.3%) were the 2 risk factors more associated with young CAD. There is a preponderance toward single vessel disease in the young CAD group, and they had a favorable clinical outcome in terms of all-cause mortality at discharge (RR 0.49 [CI 0.26-0.94]) and 1-year follow-up (RR 0.47 [CI 0.19-1.15]). We observed distinctive features of young CAD that would serve as a framework in the primary and secondary prevention of the early onset CAD.


Evaluation of a simulation-based pediatric clinical skills curriculum for medical students.
Simulation-based education is expensive and requires greater resources than traditional methods, yet there is limited evidence to justify such expenditures for medical student education. We describe the implementation and evaluation of a simulation-based curriculum delivered to medical students during a pediatric clerkship. This prospective mixed-methods study evaluated a 5-day long simulation-based clinical skills curriculum (PRE-Clerkship EDucational Exercises [PRECEDE]) at the Johns Hopkins University School of Medicine. Two hundred medical students participated in PRECEDE during a 2-year period and were compared with 236 students who had not. Outcomes were assessed across 3 levels of Kirkpatrick's framework for evaluation. The 4-level model consists of reaction, learning, behavior, and results criteria. Secondary outcomes measured changes in assessment scores across 16 student performance domains during clerkship, changes in performance on the National Board of Medical Examiners subject examination in pediatrics, and student assessments of the curriculum. Improvements were noted across 3 levels of the Kirkpatrick's model. Student performance evaluations were significantly higher across all 16 evaluation components, with effect sizes ranging from small to medium (Cohen's d, 0.23-0.44). Students scored significantly higher on the National Board of Medical Examiners pediatric shelf examination (80 vs. 77, P<0.001). Ninety-seven percent of the medical students agreed that their skills increased and that the time lost to real clinical experiences was a worthwhile trade-off for this curriculum The implementation of a simulation-based curriculum within a pediatrics clerkship resulted in higher knowledge scores and led to improvements in medical student clinical performance during the clerkship.


Oral Bacteriotherapy in Patients With COVID-19: A Retrospective Cohort Study.
Background: Mounting evidence suggests SARS-CoV-2 may impact on host microbiota and gut inflammation, infecting intestinal epithelial cells. This possible link and its implications can be investigated by observing the effects of modulation of the microbial flora in patients with COVID-19. The aim of this study was to compare the rate of mortality, the need of ICU hospitalization and the length of hospitalization in patients with severe COVID-19 pneumonia who received the best available therapy (BAT) vs. patients treated with BAT and supplemented with oral bacteriotherapy. Methods: This retrospective, observational cohort study included 200 adults with severe COVID-19 pneumonia. All patients received therapeutic regimens including low molecular weight heparin plus one or more between hydroxychloroquine, azithromycin, antivirals, and Tocilizumab. Oral bacteriotherapy was used as complementary treatment. Results: Out of the 200 patients, 112 received BAT without oral bacteriotherapy, and 88 BAT with oral bacteriotherapy. Crude mortality was 22%. Eleven percent died in the group of patients treated with BAT plus oral bacteriotherapy vs. 30% subjects in the group of patients managed only with BAT (p < 0.001). By multivariate analysis, the age >65 years, CRP >41.8 mg/L, Platelets <150.000 mmc, and cardiovascular events were associated with the increased risk of mortality. Oral bacteriotherapy was an independent variable associated with a reduced risk for death. Despite large prospective trials are needed, this study highlights a possible role for oral bacteriotherapy in the management of patients hospitalized for COVID-19 pneumonia.


A population-level strain genotyping method to study pathogen strain dynamics in human infections.
A hallmark of chronic bacterial infections is the long-term persistence of 1 or more pathogen species at the compromised site. Repeated detection of the same bacterial species can suggest that a single strain or lineage is continually present. However, infection with multiple strains of a given species, strain acquisition and loss, and changes in strain relative abundance can occur. Detecting strain-level changes and their effects on disease is challenging because most methods require labor-intensive isolate-by-isolate analyses, and thus, only a few cells from large infecting populations can be examined. Here, we present a population-level method for enumerating and measuring the relative abundance of strains called population multi-locus sequence typing (PopMLST). The method exploits PCR amplification of strain-identifying polymorphic loci, next-generation sequencing to measure allelic variants, and informatic methods to determine whether variants arise from sequencing errors or low-abundance strains. These features enable PopMLST to simultaneously interrogate hundreds of bacterial cells that are cultured en masse from patient samples or are present in DNA directly extracted from clinical specimens without ex vivo culture. This method could be used to detect epidemic or super-infecting strains, facilitate understanding of strain dynamics during chronic infections, and enable studies that link strain changes to clinical outcomes.


Effects of d-amphetamine and chlordiazepoxide on spaced responding in pigeons.
The effects of d-amphetamine and chlordiazepoxide were studied in pigeons on performance (1) under a schedule that reinforced responses on a key (food key) if they were more than 20 sec apart, (2) under the same schedule when responses also were required on a collateral key during the interresponse time on the food key, and (3) under the same schedule when responses were required on a collateral key during the interresponse time on the food key and collateral-key responses could produce a stimulus correlated with the availability of food. Under all three spaced-responding schedules, d-amphetamine and chlordiazepoxide at low dose levels slightly increased the frequency of short interresponse times on the food key for about half the birds, and either did not affect the interresponse time patterns of the other birds, or lengthened the durations slightly. At higher dose levels, d-amphetamine and chlordiazepoxide increased the frequency of long interresponse times or abolished responding in all birds. Changes in the pattern of interresponse times on the food key did not seem to depend on changes in the rate or pattern of collateral-key responses.


Clinical value of research in chronic spontaneous abortion.
We have focused on comparisons of abnormal with normal pregnancies. Humoral and cellular aspects were studied in 50 couples with unexplained consecutive pregnancy failures and compared with 15 fertile couples. The results show that spontaneous aborters generally fall into two groups: Primary spontaneous aborters are those women who, by the same husband, never have carried a pregnancy beyond 20 weeks; secondary spontaneous aborters have had, by the same husband, either a normal pregnancy or a pregnancy failure beyond the 20th week. Primary spontaneous aborting couples tend to share a significantly greater number of HLA antigens than do secondary aborters or normally fertile mates. Secondary spontaneous aborters manifest cytotoxic non-HLA-dependent antibody to husbands' lymphocytes in complement-dependent assays; these are antibodies to trophoblast-lymphocyte cross-reactive (TLX) antigens. Secondary but not primary aborters have complement-independent antibody-dependent cell-mediated cytotoxicity reactions to their husbands' lymphocytes. Both exhibit diminished mixed lymphocyte culture reactions between mating partners if the tests are done in autologous plasma; only primary aborters show poor reactions if the assays are done in control sera. Immunotherapy with nonhusband lymphocytes in more than 20 primary spontaneous aborters has resulted in greater than 90% successful subsequent pregnancies. Leukocyte immunotherapy has not been useful for secondary aborters, but two of three women have had normal pregnancies following daily subcutaneous injections of heparin. The clinical relevance of these findings will be discussed.


Occupational exposure in small and medium scale industry with specific reference to heat and noise.
This study was undertaken to assess heat and noise exposure and occupational safety practices in small and medium scale casting and forging units (SMEs) of Northern India. We conducted personal interviews of 350 male workers of these units through a comprehensive questionnaire and collected information on heat and noise exposure, use of protective equipment, sweat loss and water intake, working hour. The ambient wet bulb globe temperature (WBGT index) was measured using quest temp 34/36o area heat stress monitor. A-weighted Leq ambient noise was measured using a quest sound level meter "ANSI SI. 43-1997 (R 2002) type-1 model SOUNDPRO SE/DL". We also incorporated OSHA norms for hearing conservation which include - an exchange rate of 5dB(A), criterion level at 90dB(A), criterion time of eight hours, threshold level is equal to 80dB(A), upper limit is equal to 140dB(A) and with F/S response rate. Results of the study revealed that occupational heat exposure in melting, casting, forging and punching sections is high compared to ACGIH/NIOSH norms. Ambience noise in various sections like casting / molding, drop forging, cutting presses, punching, grinding and barreling process was found to be more than 90dB(A). About 95% of the workers suffered speech interference where as high noise annoyance was reported by only 20%. Overall, 68% workers were not using any personal protective equipment (PPE). The study concluded that the proportion of SME workers exposed to high level heat stress and noise (60 - 72 hrs/week) is high. The workers engaged in forging and grinding sections are more prone to noise induced hearing loss (NIHL) at higher frequencies as compared to workers of other sections. It is recommended that there is a strong need to implement the standard of working hours as well as heat stress and noise control measures.


Rapid diagnostic techniques for bacterial meningitis.
The purpose of this review is to provide an overview of the most prevalent current methods for rapid diagnosis of meningitis. In addition, some of those techniques which, while not yet practicable for most laboratories but are on the clinical horizon, have been surveyed. Two principles which emerge from this review are that many of the older techniques, such as Gram stain, retain their clinical utility by virtue of their practicality, cost-benefit ratio, and simplicity; and many of the more sophisticated techniques which are now emerging from the research arena (such as FPEC-GLC), facilitate rapid diagnosis where it has not previously existed. Also, when identification of the infectious agent depends on antigen detection, prior treatment will not compromise the results. Therefore, the combination of old and new laboratory methods for diagnosing meningitis provides a much wider, more effective and efficient set of diagnostic tools than was previously available.


Paliperidone: As a Possible Cause of Pericardial Effusion.
Paliperidone which is the active metabolite of risperidon is one of the novel antipsychotics. Controlled clinical studies proved its effectiveness on decreasing the positive and negative symptoms of schizophrenia. Some studies and case reports, reported its cardiovascular and cardiometabolic side effects. Cardiac tamponade, that may be mortal, wasn't reported as a side effect with paliperidone usage before. We present a 40 years old female with catatonic schizophrenia for eighteen years, whom was on 6 mg/day paliperidon for 3 years and presented with pericardial tamponade. We suggest that paliperdione may have cardiovascular and cardiometabolic side effects also in therapeutic doses.


C-reactive protein testing in patients with acute rhinosinusitis leads to a reduction in antibiotic use.
To evaluate the effect of C-reactive protein (CRP) testing on the antibiotic prescribing in patients with acute rhinosinusitis. Audit-based study carried out in primary care centres in Spain. GPs registered episodes of rhinosinusitis during 3-week period before and after an intervention. Two types of intervention were considered: full intervention group (FIG) consisting in individual feedback based on results from the first registry, courses in rational antibiotic prescribing, guidelines, patient information leaflets, workshops on rapid tests and use of the CRP test. GPs in the partial intervention group (PIG) underwent all the above intervention except for the workshop and they did not have access to CRP. Multilevel logistic regression analysis was performed considering the prescription of antibiotics as the dependent variable. Two hundred and ten physicians were assigned to FIG and 71 to PIG. In 2009, 59 new physicians were included as a control group. Two hundred and sixty-seven GPs visited contacts with rhinosinusitis (78.5%) registering a total of 836 cases. In the group of GPs with access to CRP rapid test, 207 patients with rhinosinusitis (75.3%) were tested and antibiotics were prescribed in 156 patients (56.7%). Antibiotics were prescribed in 87 patients (82.9%) in the group of GPs exposed to PIG and in 52 patients (86.7%) in the control group (P < 0.001). Antibiotic prescription was significantly reduced after the intervention among physicians assigned to FIG, with an odds ratio of antibiotic prescribing of 0.12 (95% confidence interval: 0.01-0.32). Physicians with access to CRP tests significantly reduced antibiotic prescription in patients with rhinosinusitis.


Rapid screening method for simultaneous determination of four polycyclic aromatic hydrocarbons in water samples by derivative non-linear variable-angle synchronous fluorescence spectrometry.
Derivative non-linear variable-angle synchronous fluorescence spectroscopy (D-NLVASFS) has been developed to improve the selectivity of the fluorescence measurement without loss of sensitivity. We report a simple screening approach for the rapid and simultaneous determination of 1,2.5,6-dibenzoanthracene (DBA), 2,3-benzofluorene (BF), pyrene (Pyr) and 3,4-benzopyrene (BaP) in a mixture by using first-derivative NLVASFS. The method is efficient, fast and straightforward, and the measurement was simply based on a single spectrum via a single spectral scanning of a sample. The linear ranges 0.005-0.30, 0.02-0.30, 0.02-0.40 and 0.005-1.0 microg/mL and the detection limits 0.08, 1.14, 1.64 and 0.12 ng/mL are obtained for DBA, BF, Pyr and BaP, respectively. The method was applied to water samples spiked with the four polycyclic aromatic hydrocarbons (PAHs), with mean recoveries of 103.0% (SD 5.9%) for DBA, 96.4% (SD 4.2%) for BF, 96.2% (SD 4.0%) for Pyr and 98.2% (SD 3.0%) for BaP.


Deep Fusion Network Based Sparse View CT Reconstructions for Clinical Diagnostic Scanners.
Sparse view CT scan has the advantage of reducing radiation exposure and scanning time in clinical diagnosis. However, the limited number of x-ray projections can make the reconstruction problem ill posed and result in image artifacts. To tackle the problem, we propose a novel model-based deep fusion network(DFN) satisfying the clinical set-up. It extracts fused features encoded from both the sinogram and the preliminary reconstructed image generated by filtered back projection (FBP) to improve the quality of reconstruction. The preliminary reconstructed image endows fused features with prior knowledge that facilitate the convergence of neural network to high-quality reconstruction images. We design a custom loss for training that enforces the network to learn both the pixel value and the integrity of the tissue structure. A synthetic sparse view breast CT dataset from American Association of Physicists in Medicine(AAPM) is used for training, validation and testing. The qualitative and quantitative evaluations show that the DFN reconstruction algorithm significantly improves in balancing between the image quality and reconstruction speed, hence enables fast and high quality CT reconstruction despite the sparse view limitations.


Massive congenital intracranial immature teratoma of the lateral ventricle with retro-orbital extension: a case report and review of the literature.
Germ cell tumors comprise 0.4-3.1% of all intracranial tumors, and teratoma constitutes 9-30% of them. Congenital intracranial tumors are very rare and only account for 0.5-1.5% of all childhood brain tumors. The most common type of these tumors present at birth is teratomas, which represent 0.5% of all intracranial tumors. Most teratomas are midline tumors located predominantly in the sellar and pineal regions. In this study, we report a neonatal intracranial immature teratoma at the lateral ventricle because of its rare location. A 5-day-old female neonate presented with a history of irritability and left eye protrusion since birth. A head computed tomographic (CT) scan and magnetic resonance imaging (MRI) disclosed a large tumor filling the left lateral ventricle and extending into the ipsilateral retro-orbital space. With left frontotemporoparietal craniotomy, a large grayish-white lobulated vascular mass was encountered, and total removal of tumor was performed. Histological examination revealed the diagnosis of immature teratoma. The prognosis of congenital intracranial immature teratoma is usually poor because the lesions are extensive when they are identified. Prenatal ultrasonography is necessary for the prenatal diagnosis. Fetal MRI should be made for the evaluation of intracranial tumor. If the tumor is detected before the 24th week of gestation, termination of the pregnancy should be considered.


Small bowel feeding tube placement using an electromagnetic tube placement device: accuracy of tip location.
An electromagnetic tube placement device (ETPD) monitors tip position of feeding tubes (FT) during placement in the digestive tract. It helps to avoid airway misplacement and permits positioning into the small bowel (SB). This study compares the overall agreement between FT tip location as determined by an ETPD vs an abdominal radiograph of the kidneys, ureter, and bladder (KUB). Using an ETPD, A nurse placed postpyloric FTs in ICU patients. We included all patients in whom the ETPD was used for FT placement. Data were prospectively recorded for 255 days on the rate of successful postpyloric placement, ETPD estimated tip location, and KUB location. 860 tubes were placed in 616 patients, 719 (83.6%) of which recorded for ETPD and KUB. According to the KUB, 81% of tubes were in the SB; however, ETPD suggested 89% were beyond the pylorus. There was moderate agreement beyond what could be attributed to chance between KUB and ETPD tip locations (475 [66.1%], κ score 0.62 [95% confidence interval 0.58-0.67]). More tubes by KUB were distal (134[18.6%]) vs proximal (110[15.3%]) to the suspected location by ETPD (P < .0001. Tubes in or distal to the second half of the duodenum, according to ETPD were rarely in the stomach (<1%). No tubes were proximal to the stomach or placed into the airway. The strong agreement between KUB and ETPD, when tubes were believed to be in the second part of the duodenum or beyond, suggests that KUB is necessary only when the FT tip is suspected to be in the proximal duodenum.


Biomechanical Forces and Oxidative Stress: Implications for Pulmonary Vascular Disease.
Significance: Oxidative stress in the cell is characterized by excessive generation of reactive oxygen species (ROS). Superoxide (O2-) and hydrogen peroxide (H2O2) are the main ROS involved in the regulation of cellular metabolism. As our fundamental understanding of the underlying causes of lung disease has increased it has become evident that oxidative stress plays a critical role. Recent Advances: A number of cells in the lung both produce, and respond to, ROS. These include vascular endothelial and smooth muscle cells, fibroblasts, and epithelial cells as well as the cells involved in the inflammatory response, including macrophages, neutrophils, eosinophils. The redox system is involved in multiple aspects of cell metabolism and cell homeostasis. Critical Issues: Dysregulation of the cellular redox system has consequential effects on cell signaling pathways that are intimately involved in disease progression. The lung is exposed to biomechanical forces (fluid shear stress, cyclic stretch, and pressure) due to the passage of blood through the pulmonary vessels and the distension of the lungs during the breathing cycle. Cells within the lung respond to these forces by activating signal transduction pathways that alter their redox state with both physiologic and pathologic consequences. Future Directions: Here, we will discuss the intimate relationship between biomechanical forces and redox signaling and its role in the development of pulmonary disease. An understanding of the molecular mechanisms induced by biomechanical forces in the pulmonary vasculature is necessary for the development of new therapeutic strategies.


It's big surgery: preoperative expressions of risk, responsibility, and commitment to treatment after high-risk operations.
To identify the processes, surgeons use to establish patient buy-in to postoperative treatments. Surgeons generally believe they confirm the patient's commitment to an operation and all ensuing postoperative care, before surgery. How surgeons get buy-in and whether patients participate in this agreement is unknown. We used purposive sampling to identify 3 surgeons from different subspecialties who routinely perform high-risk operations at each of 3 distinct medical centers (Toronto, Ontario; Boston, Massachusetts; Madison, Wisconsin). We recorded preoperative conversations with 3 to 7 patients facing high-risk surgery with each surgeon (n = 48) and used content analysis to analyze each preoperative conversation inductively. Surgeons conveyed the gravity of high-risk operations to patients by emphasizing the operation is "big surgery" and that a decision to proceed invoked a serious commitment for both the surgeon and the patient. Surgeons were frank about the potential for serious complications and the need for intensive care. They rarely discussed the use of prolonged life-supporting treatment, and patients' questions were primarily confined to logistic or technical concerns. Surgeons regularly proceeded through the conversation in a manner that suggested they believed buy-in was achieved, but this agreement was rarely forged explicitly. Surgeons who perform high-risk operations communicate the risks of surgery and express their commitment to the patient's survival. However, they rarely discuss prolonged life-supporting treatments explicitly and patients do not discuss their preferences. It is not possible to determine patients' desires for prolonged postoperative life support on the basis of these preoperative conversations alone.


Female acceleration tolerance: effects of menstrual state and physical condition.
The literature contains a paucity of information on female tolerance to high sustained acceleration. With women now flying high-performance aircraft, gender-specific factors that may affect female acceleration tolerance have become increasingly important. The purpose of this investigation was to determine how menstrual state and physical condition affect acceleration tolerance. We hypothesized the menstrual cycle would have no effect on acceleration tolerance and that a positive correlation would exist between physical fitness level and tolerance to high sustained acceleration. Centrifuge exposures on 8 female subjects consisted of a relaxed gradual-onset run (0.1 G.s-1) to the visual endpoint, a rapid-onset run (6 G.s-1) to +5 GZ for 15 s, and a +4.5 to +7 GZ simulated aerial combat maneuver (SACM) to physical exhaustion. Acceleration tolerance data were collected at onset of menstruation and 1, 2 and 3 weeks following the onset for two complete menstrual cycles. On separate days, body composition, anaerobic power output and peak oxygen uptake were determined. Retrospective data from 10 male subjects who had performed the +4.5 to +7 GZ SACM were analyzed and compared to these data. Analysis of variance revealed no significant difference in relaxed tolerance or SACM duration between the four selected menstrual cycle time points. Time-to-fatigue on the +4.5 to +7 GZ SACM was positively (p < or = 0.05) correlated with absolute fat-free mass (r = 0.87) and anaerobic power production (r = 0.76) in female subjects. However, when these variables were adjusted for total body mass, the significant correlations no longer existed. No correlation was found between SACM duration and absolute (L min-1) nor relative (ml.kg-1.min-1) aerobic fitness. Time-to-fatigue during the SACM was not significantly different between male and female subjects (250 +/- 97 and 246 +/- 149 s, respectively).


Efficacy of Recombinant Human Thrombopoietin for the Treatment of Secondary Failure of Platelet Recovery After Allogeneic HSCT.
Secondary failure of platelet recovery (SFPR) is a life-threatening complication that may affect up to 20% of patients after allogeneic hematopoietic stem cell transplantation (HSCT). In this study, to evaluate the efficacy of recombinant human thrombopoietin (rhTPO), we retrospectively analyzed 29 patients who received continuous rhTPO for the treatment of SFPR. Overall response and complete response were observed in 24 (82.8%) patients and 10 (34.5%) patients, at a median time of 21.5 days (range, 3-41 days) and 39.5 days (range, 7-53 days) after initiation of rhTPO treatment, respectively. Among the responders, the probability of keeping overall response and complete response at 1 year after response was 77.3% and 80.0%, respectively. In multivariate analysis, higher CD34+ cells (≥3 × 106/kg) infused during HSCT (HR: 7.22, 95% CI: 1.53-34.04, P = 0.01) and decreased ferritin after rhTPO treatment (HR: 6.16, 95% CI: 1.18-32.15, P = 0.03) were indicated to associate with complete response to rhTPO. Importantly, rhTPO was well tolerated in all patients without side effects urging withdrawal and clinical intervention. The results of this study suggest that rhTPO may be a safe and effective treatment for SFPR.


Recruitment of minority and underserved populations in the United States: the Centers for Population Health and Health Disparities experience.
The recruitment of minority and underserved individuals to research studies is often problematic. The purpose of this study was to describe the recruitment experiences of projects that actively recruited minority and underserved populations as part of The Centers for Population Health and Health Disparities (CPHHD) initiative. Principal investigators and research staff from 17 research projects at eight institutions across the United States were surveyed about their recruitment experiences. Investigators reported the study purpose and design, recruitment methods employed, recruitment progress, problems or challenges to recruitment, strategies used to address these problems, and difficulties resulting from Institutional Review Board (IRB) or Health Insurance Portability and Accountability Act of 1996 (HIPAA) requirements. Additionally, information was collected about participant burden and compensation. Burden was classified on a three-level scale. Recruitment results were reported as of March 31, 2007. Recruitment attainment ranged from 52% to 184% of the participant recruitment goals. Commonly reported recruitment problems included administrative issues, and difficulties with establishing community partnerships and contacting potential participants. Long study questionnaires, extended follow-up, and narrow eligibility criteria were also problematic. The majority of projects reported difficulties with IRB approvals, though few reported issues related to HIPAA requirements. Attempted solutions to recruitment problems varied across Centers and included using multiple recruitment sites and sources and culturally appropriate invitations to participate. Participant burden and compensation varied widely across the projects, however, accrual appeared to be inversely associated with the amount of participant burden for each project. Recruitment of minority and underserved populations to clinical trials is necessary to increase study generalizbility and reduce health disparities. Our results demonstrate the importance of flexible study designs which allow adaptation to recruitment challenges. These experiences also highlight the importance of involving community members and reducing participant burden to achieve success in recruiting individuals from minority and underserved populations.


Increasing methane content in biogas and simultaneous value added product recovery using microbial electrosynthesis.
Electrosynthesis of multi-carbon compounds from the carbon dioxide present in biogas is a nascent approach towards purification of biogas. Microbial electrosynthesis (MES) cells, fabricated using different electrode materials, were operated using different electrolytes and mixed anaerobic culture as biocatalysts in the cathodic chamber under an applied cathode potential of -0.7 V vs standard hydrogen electrode (SHE). The rate of production of acetate, isobutyrate, propionate and 2-piperidinone from reduction of CO2 in the cathodic chamber of the MES was 0.81 mM/day, 0.63 mM/day, 0.44 mM/day and 0.53 mM/day, respectively. As methane was also present in the biogas, methyl derivatives of these acids were also found in traces in catholyte. It was observed that the use of nickel foam as an anode, 1 M NiSO4 solution as anolyte, graphite felt as a cathode, phosphate buffer solution as catholyte at a pH of 5.2 proved to be the best possible combination for MES for this study to get enhanced product yield at higher energy efficiency.


Identification of conserved domains in Salmonella muenchen flagellin that are essential for its ability to activate TLR5 and to induce an inflammatory response in vitro.
The bacterial surface protein flagellin is widely distributed and well conserved among distant bacterial species. We and other investigators have reported recently that purified flagellin from Salmonella dublin or recombinant flagellin of Salmonella muenchen origin binds to the eukaryotic toll receptor TLR5 and activates the nuclear translocation of NF-kappaB and mitogen-activated protein kinase, resulting in the release of a host of pro-inflammatory mediators in vitro and in vivo. The amino acid sequence alignment of flagellins from various Gram-negative bacteria shows that the C and N termini are well conserved. It is possible that sequences within the N and C termini or both may regulate the pro-inflammatory activity of flagellin. Here we set out to map more precisely the regions in both termini that are required for TLR5 activation and pro-inflammatory signaling. Systematic deletion of amino acids from either terminus progressively reduced eukaryotic pro-inflammatory activation. However, deletion of amino acids 95-108 (motif N) in the N terminus and 441-449 (motif C) in the C terminus abolished pro-inflammatory activity completely. Site-directed mutagenesis analysis provided further evidence for the importance of motifs N and C. We also present evidence for the functional role of motifs N and C with the TLR5 receptor using a reporter assay system. Taken together, our results demonstrate that the pro-inflammatory activity of flagellin results from the interaction of motif N with the TLR5 receptor on the cell surface.


Development of dermal connective tissue in human embryonic and fetal skin.
The primordial dermis begins as a cellular mesenchyme and transforms into a fibrous, sparsely cellular network. At six weeks gestation a fine meshwork of collagen fibrils underlies the dermal-epidermal junction and adheres to the dermal mesenchymal cell surfaces. With increasing gestational age, the amount of extracellular collagen matrix increases, fibrils associate into collagen fiber bundles, cells are spaced further apart and their elongated axes tend to orient parallel to the plane of the skin. The fine collagen meshwork persists throughout development at the dermal-epidermal junctions and ensheathes epidermal appendages as they extend into the dermis. A similar network encases developing vessels. The overall thickness of the dermis increases from 0.1 to 0.15 mm at 7 weeks to greater that 0.7 mm at 20 weeks. By 16 weeks gestation the epidermal appendages extend deeply into the dermis and the dermis is organized into two regions: an upper, finely fibrillar papillary region and a deep reticular region that contains larger collagen fiber bundles. A vascular plexus separates the dermis from the hypodermis. The dermis of the 20-week fetus is adult-like in overall appearance, although it is still smaller in total thickness and size of collagen fibrils and fiber bundles than in the newborn skin.


Transforming growth factor-beta protection of cancer cells against tumor necrosis factor cytotoxicity is counteracted by hyaluronidase (review).
Numerous cancer cells, when exposed to transforming growth factor beta (TGF-beta), become resistant to tumor necrosis factor (TNF) cytotoxicity. Pretreatment of L929 fibroblasts, for example, with TGF-beta isoforms (beta 1, beta 2 and beta 3) for at least 0.5-1 h results in resistance to TNF killing. TGF-beta 1 mediates the following sequential events in L929 cells: i) rapid induction of protein tyrosine-phosphorylation (< 30 min), ii) stimulation of protective protein synthesis and acquisition of TNF resistance (approximately 0.5-1 h), and iii) suppression of I kappa B-alpha expression (1-2 h). Two protective proteins induced by TGF-beta 1 are a 46 kDa extracellular matrix TNF-resistance triggering (TRT) protein and a putative transmembrane anti-apoptotic adhesion protein TIF2 (containing and RGD motif in the extracellular region). Both proteins enable L929 cells to resist TNF killing. Notably, testicular hyaluronidase increases TNF sensitivity in several types of cancer cells, counteracts TGF-beta-mediated TNF-resistance, and suppresses TGF-beta 1 gene expression in L929 cells in a serum-dependent manner. Moreover, hyaluronidase antagonizes TGF-beta-mediated inhibition of epithelial cell growth. Both TGF-beta and hyaluronidase are essential for the progression and invasiveness of breast, prostate and other cancers. Conceivably, a stage-dependent expression, as well as a balanced production, of these proteins is essential for cancer development and self protection against TNF cytotoxicity.


Radiation Induces Valvular Interstitial Cell Calcific Response in an in vitro Model of Calcific Aortic Valve Disease.
Background: Mediastinal ionizing radiotherapy is associated with an increased risk of valvular disease, which demonstrates pathological hallmarks similar to calcific aortic valve disease (CAVD). Despite advances in radiotherapy techniques, the prevalence of comorbidities such as radiation-associated valvular disease is still increasing due to improved survival of patients receiving radiotherapy. However, the mechanisms of radiation-associated valvular disease are largely unknown. CAVD is considered to be an actively regulated disease process, mainly controlled by valvular interstitial cells (VICs). We hypothesize that radiation exposure catalyzes the calcific response of VICs and, therefore, contributes to the development of radiation-associated valvular disease. Methods and Results: To delineate the relationship between radiation and VIC behavior (morphology, calcification, and matrix turnover), two different in vitro models were established: (1) VICs were cultured two-dimensional (2D) on coverslips in control medium (CM) or osteogenic medium (OM) and irradiated with 0, 2, 4, 8, or 16 Gray (Gy); and (2) three-dimensional (3D) hydrogel system was designed, loaded with VICs and exposed to 0, 4, or 16 Gy of radiation. In both models, a dose-dependent decrease in cell viability and proliferation was observed in CM and OM. Radiation exposure caused myofibroblast-like morphological changes and differentiation of VICs, as characterized by decreased αSMA expression. Calcification, as defined by increased alkaline phosphatase activity, was mostly present in the 2D irradiated VICs exposed to 4 Gy, while after exposure to higher doses VICs acquired a unique giant fibroblast-like cell morphology. Finally, matrix turnover was significantly affected by radiation exposure in the 3D irradiated VICs, as shown by decreased collagen staining and increased MMP-2 and MMP-9 activity. Conclusions: The presented work demonstrates that radiation exposure enhances the calcific response in VICs, a hallmark of CAVD. In addition, high radiation exposure induces differentiation of VICs into a terminally differentiated giant-cell fibroblast. Further studies are essential to elucidate the underlying mechanisms of these radiation-induced valvular changes.


Clinical roundtable monograph: New data in emerging treatment options for chemotherapy-induced nausea and vomiting.
Chemotherapy-induced nausea and vomiting (CINV) has long been one of the most troublesome adverse effects of chemotherapy, leading to significant detriments in quality of life and functioning, increased economic costs, and, in some cases, the discontinuation of effective cancer therapy. The past 2 decades have witnessed a dramatic increase in the number of effective antiemetic agents, with the introduction of the serotonin (5-hydroxytryptamine [5-HT₃]) receptor antagonists (ondansetron, granisetron, and palonosetron), the neurokinin-1 (NK₁) receptor antagonists (aprepitant and fosaprepitant), and the identification of other agents that have demonstrated efficacy against CINV, including corticosteroids. These agents often provide excellent control of emesis. Nausea, however, has proven more intractable, particularly in the days after administration of chemotherapy. Newer antiemetic agents under study may provide additional CINV control, particularly against delayed nausea. New agents undergoing review by the US Food and Drug Administration for the prevention of CINV include the novel NK₁ receptor antagonist rolapitant and a fixed-dose combination consisting of the novel NK₁ receptor antagonist netupitant and palonosetron (NEPA). Adherence to clinical practice guidelines has been shown to significantly improve CINV control. As antiemetic therapy continues to evolve, it will be important for clinicians to stay informed of new developments and changes in guidelines.


The effects of noise exposure on hippocampal cognition in C57BL/6 mice via transcriptomics.
Noise is an important environmental stressor in the industrialized world and has received increasing attention in recent years. Although epidemiological research has extensively demonstrated the relationship between noise and cognitive impairment, the specific molecular mechanisms and targets remain to be fully explored and understood. To address this issue, 5-month-old C57BL/6 mice were divided into two groups, with one group exposed to white noise at 98 dB. The effects of noise on cognition in mice were investigated through molecular biology and behavioral experiments. Subsequently, transcriptomic sequencing of the hippocampus in both groups of mice was performed and enrichment analysis of differentially expressed genes (DEGs) was conducted using KEGG and GO databases. Furthermore, LASSO analysis was used to further narrow down the relevant DEGs, followed by enrichment analysis of these genes using KEGG and GO databases. The DEGs were further validated by rt-qPCR. Following noise exposure, the hippocampus levels of inflammation-related factors increased, the phosphorylation of Tau protein increased, the postsynaptic density protein decreased, the number of Nissl bodies decreased, and cell shrinkage in the hippocampus increased. Moreover, the behavioral experiments manifest characteristics indicative of a decline in cognitive.A total of 472 DEGs were identified through transcriptomic analysis, and seven relevant genes were screened by the LASSO algorithm, which were further validated by PCR to confirm their consistency with the omics results. In conclusion, noise exposure affects cognitive function in mice through multiple pathways, and the omics results provide new evidence for the cognitive impairment induced by noise exposure.


Modulation of Energy Intake and Expenditure Due to Habitual Physical Exercise.
The 20th and 21st centuries are marked by an increase in life expectancy on one hand and on the other hand by the increase of so called civilization diseases. Their share one common trait: the energy metabolism imbalance, with low energy expenditure and high energy uptake. Our age can be viewed as the age of inactivity and wealth. The aim of the present review is to highlight the influence of habitual physical activity on energy metabolism and balance. Energy balance is the difference between energy intake and energy expenditure, where energy expenditure further divides into resting metabolic rate, thermic effect of feeding and energy used by physical activity. In general population, resting metabolic rate remains constant and proportional to muscle body mass. Muscle mass increases with exercise, especially resistance exercise, concomitantly with increasing energy expenditure. The effect of exercise on appetite is very strong, proportional to exercise intensity. An acute bout of aerobic exercise suppresses appetite by decreasing ghrelin plasma levels, and increasing gut hormones. Different subgroups of patients respond differently to the same exercise or habitual activity and have thus distinct effects on energy balance. Different myokines plasma levels after exercise could explain these different reactions although most of their effect is still unclear. Physical activity plays an important role in the prevention and treatment of many disorders, like obesity, type 1 and type 2 diabetes, dyslipidaemia, hypertension, coronary heart disease, osteoporosis, psychiatric and neurologic disorders. It is evident, that physical activity has an effect not only on energy balance but also has a direct effect on other body organ via its own molecules - myokines. The pharmacological effect of myokines gives hope that one day we could have a "myokine drug" that could be used in patients who are unable to exercise. Until then we should use our "muscle-pharmacy" and try to convince also our patients to use theirs. Physical activity makes a huge impact on human health.


[Dynamics of the interaction of monoclonal antibodies with antigens immobilized on plates].
A new method to determine a rate constant and total reaction product using data of reaction product kinetics is proposed. The method is based on the solution of the system of irrational equations (for special case), which describes such processes. This method was used to determine the reaction rate constant of interaction of some monoclonal antibodies with the antigens immobilized on immunological plates.


CORONARY ARTERY CALCIUM IS ASSOCIATED WITH LPA GENE VARIANT RS7765803-C IN MEXICAN MESTIZO POPULATION. THE GEA PROJECT.
Lipoprotein(a) [Lp(a)] levels are genetically determined; high levels are a risk factor for coronary disease, although their association with coronary artery calcium (CAC) is controversial. Objective: The objective of the study was to assess the association of LPA gene polymorphisms with CAC in a Mexican Mestizo population. We included 1594 subjects 35-70 years old. Six polymorphisms of the LPA gene were analyzed. CAC score was determined by tomography and Lp(a) serum levels by immunonephelometry. The association of LPA polymorphism with CAC and Lp(a) was evaluated by logistic regression. The prevalence of Lp(a) ≥30 mg/dL was 10%, and of CAC >0 was 26.9%. Three polymorphisms were associated with high Lp(a) levels: rs10455872-G (p = 0.013), rs6907156-T (p = 0.021), and rs7765803-C (p = 0.001). Homozygotes (CC) for the rs7765803 variant compared with the G allele (CG + GG) carriers had higher Lp(a) levels (8.9 [3.3-23.9] vs. 4.9 [2.3-11.2] mg/dL; p = 0.015) and higher prevalence of CAC >0 (36.5% vs. 26.3%, p = 0.045) and were associated with CAC > 0 (odds ratio = 1.7, 95% confidence interval: 1.06-2.7; p < 0.026). The other polymorphisms were not associated with CAC. This is the first study to demonstrate in a Mexican Mestizo population that carriers of the rs7765803-C allele of LPA gene have 2.6 times greater risk for high Lp(a) values and 1.7 times higher risk for coronary artery disease.