Derivatization-assisted LC-MS/MS method for simultaneous quantification of endogenous gamma-hydroxybutyric acid and its metabolic precursors and products in human urine.
The accurate, precise, and robust quantification of endogenous biomarkers is a challenging task because of the presence of significantly low levels of endogenous compounds in biological samples, the absence of analyte-free matrix-matched calibrators, and sample instability due to in-vitro production or degradation of the analytes. Gamma-hydroxybutyric acid (GHB), a compound often used in drug-facilitated crimes, is a human neurotransmitter produced during both the biosynthesis and metabolism of gamma-aminobutyric acid (GABA). Evidently, proving GHB intoxication through the quantification of GHB and its metabolites in biological samples is not straightforward. This study aimed to develop a sensitive and accurate quantitative method for the simultaneous determination of endogenous GHB and its metabolic precursors and products (glutamic acid, GABA, succinic acid, 2,4-dihydroxybutyric acid, 3,4-dihydroxybutyric acid, glycolic acid, and succinylcarnitine) in human urine using LC-MS/MS. For this purpose, chemical derivatization with benzoyl chloride was employed to improve the sensitivity to glutamic acid and GABA. Synthetic urine was used to prepare calibrators, and the validity of this approach was fully demonstrated, particularly focusing on the instability issues. The validation results proved the method to be selective, sensitive, accurate, and precise, with acceptable linearity within calibration ranges. Moreover, our results regarding the in-vitro production or degradation of metabolites highlight the effects of handling and storage conditions of urine samples. Finally, this effective analytical method is expected to be useful in studying the relationship between GHB intoxication and metabolic alterations and, thus, discovering practical biomarkers for GHB ingestion.


Taking health care to factory floor proves smart move for growing Ontario company.
A fully equipped, state-of-the-art wellness centre that employs physicians and other health care providers to meet the health needs of more than 1200 employees is being credited with giving a Canadian company a leg up on the competition.


Mesenchymal stem cells for bone repair: preclinical studies and potential orthopedic applications.
Mesenchymal stem cells (MSCs), derived from adult bone marrow, are multi-potent stem cells capable of differentiating along several lineage pathways. From a small bone marrow aspirate, MSCs can be readily isolated and easily expanded. Therefore, MSCs are thought to be a readily available source of cells for many tissue engineering and regenerative medicine applications. This review covers preclinical models that evaluate the efficacy of MSC-loaded scaffolds in large bone defects as a potential substitute for autologous and allogeneic bone grafts. This review also covers new approaches to clinical use of MSC technology.


Old age-associated enrichment of peripheral T regulatory cells and altered redox status in pulmonary tuberculosis patients.
Aging influences the susceptibility and prognosis to various infectious diseases including tuberculosis (TB). Despite the impairment of T-cell function and immunity in older individuals, the mechanism for the higher incidence of TB in the elderly remains largely unknown. Here, we evaluated the age-associated immune alterations, particularly in effector and Treg responses in pulmonary TB patients. We also evaluated the impact of redox status and its modulation with N-acetyl-cysteine (NAC) in elderly TB. Higher frequency of Treg cells and reduced IFN-γ positive T cells were observed among older TB patients. The elevated number of Treg cells correlated tightly with bacillary load (i.e. disease severity); which declined significantly in response to successful anti-tubercular treatment. We could rescue Myobacterium tuberculosis-specific effector T cell (Th1) responses through various in vitro approaches, for example, Treg cell depletion and co-culture experiments, blocking experiments using antibodies against IL-10, TGF-β, and programmed death-1 (PD-1) as well as NAC supplementation. We report old age-associated enrichment of Treg cells and suppression of M. tuberculosis-specific effector T (Th1) cell immune responses. Monitoring these immune imbalances in older patients may assist in immune potentiation through selectively targeting Treg cells and/or optimizing redox status by NAC supplementation.


Oxidative stress in neurodegeneration in dentatorubral-pallidoluysian atrophy.
Dentatorubral-pallidoluysian atrophy (DRPLA) is one of the CAG-repeat diseases, and is classified into juvenile and early adult types showing progressive myoclonus epilepsy (PME) in addition to late adult type. We immunohistochemically examined accumulation of oxidative products and expression of superoxide dismutase (SOD) in autopsy cases of DRPLA. Oxidative products to nucleosides, 8-hydroxy-2'-deoxyguanosine and 8-hydroxyguanosine, were accumulated in the lenticulate nucleus predominantly in DRPLA cases having PME. Neuronal accumulation of 4-hydroxy nonenal, a reactive lipid aldehyde, was found in the hippocampus, globus pallidus and cerebellar dentate nucleus in adult DRPLA cases and controls. Cytoplasmic immunoreactivity for Cu/ZnSOD was reduced in the external segment of globus pallidus, dentate nucleus and cerebellar cortex in DRPLA cases. Mitochondrial immunoreactivity for MnSOD was reduced in the lenticulate nucleus and cerebellum in DRPLA cases having PME. Some DRPLA cases showed reduced immunoreactivity for MnSOD in the cerebral cortex. Coexistence of reduced SOD expression and polyglutamine was observed in a few cases. It has been discussed in Huntington's disease that expanded polyglutamine can lead to oxidative neurodegeneration. It is likely that oxidative stress can be involved in DRPLA, although relationship with expanded polyglutamine remains to be elusive.


Blastocyst cryopreservation and cryopreservation-warming transfer is an effective embryo transfer strategy for day 1 rescue intracytoplasmic sperm injection cycles.
This was a retrospective analysis of a total of 625 r-ICSI cycles using freeze-all-embryos and embryo transfers (ET) in subsequent cryopreservation-warming cycles to determine the effect of the ET method for day 1 rescue intracytoplasmic sperm injection cycles (r-ICSI). Two methods were used: in method 1, cleavage-stage embryos were frozen and were directly transferred in a subsequent cryopreservation-warming cycle (r-ICSI frozen cleavage), and 144 cleavage-stage ETs occurred. Similarly, in method 2, there were 188 blastocyst-stage ETs (r-ICSI frozen blast) performed. The live birth rate (LBR) for r-ICSI frozen blast was better than that for r-ICSI frozen cleavage in calculation of ET cycles (19.44% vs. 37.77%) and also remained better after the use of logistic regression analysis (OR = 2.721, 95% CI 1.604-4.616). Conservative cumulative LBR were compared between r-ICSI frozen cleavage and r-ICSI frozen blast with regard to oocyte retrieval cycles (17.39% vs. 15.30%). The same results were obtained for conservative cumulative LBR after logistic regression analysis (OR = 0.925, 95% CI 0.557-1.535). The results of this study confirmed that it was valuable to perform r-ICSI if using freeze-all-embryos. Further, r-ICSI embryos were cultured to blastocyst stage, cryopreserved, and used in subsequent cryopreservation-warming cycles, which was an effective embryo transfer strategy and obtained satisfactory results.


Copper distribution and hydrolase activities in a contaminated soil amended with dolomitic limestone and compost.
Chemical fractionation of copper in bulk soil and its distribution in the particle-size fractions were analyzed in a Cu-contaminated soil (674 ± 122 μg Cu g(-1), up to 1900 μg Cu g(-1) in the clay fraction) sampled from a wood preservation site left untreated and subsequently treated with dolomitic limestone (DL, 0.2% w/w) and compost (CM, 5% w/w), singly and in combination (DL+CM). Soil enzymatic activities of leucine aminopeptidase, cellulase, N-acetyl-β-glucosaminidase, arylsulfatase, β-glucosidase, acetate esterase, butyric esterase, and acid phosphatase were determined. Chemical speciation showed that Cu was mostly present in the acid-soluble and reducible fractions in both untreated and treated soils, whereas treatments with DL and CM reduced the soluble and exchangeable Cu fractions, due to Cu precipitation and complexation, and increased Cu bound to soil organic matter. Analysis of the particle-size fractions showed that more than 80% of Cu was in the silt and clay fractions and that treatment with CM increased the concentration of Cu in the sand size fractions. Soil treatment with DL and CM, singly or in combination, increased hydrolase activities, mainly in the clay fraction, with the largest positive effects on N-acetyl-β-glucosaminidase, leucine aminopeptidase, and β-glucosidase activities. Overall, results confirm that (1) Cu in contaminated soils is mainly bound to the silt-clay fraction, (2) CM additions change its allocation in the particle-size fractions, and (3) treatments with DL and CM singly and in combination reduce Cu solubility and its inhibitory effects on soil enzyme activities.


Case report: A surgically treated case of diaphragmatic clear cell carcinoma without relation to endometriosis.
Diaphragmatic tumor is a rare neoplastic disease. Only three reports have revealed diaphragmatic primary clear cell carcinoma. On the other hand, secondary membranous nephropathy is sometimes triggered by the carcinoma. We describe a case of primary diaphragmatic clear cell carcinoma without relation to endometriosis or ovarian malignancies, and secondary membranous nephropathy was triggered by diaphragmatic primary clear cell carcinoma. A 67-year-old woman was found to have membranous nephropathy due to examination for renal dysfunction. A rare diaphragmatic tumor was identified on CT scan for rule out secondary membranous nephropathy. She had underwent resection of the right diaphragm tumor and reconstruction with expanded polyterafluoroethylene (e-PTFE). Pathological examination revealed the presence of clear cells with papillary arrangement and no findings of the endometriosis. Immunohistochemistry revealed that the tumor was positive for CK7, p53, and HNF-1-beta. And there had been no evidence of ovarian malignancies. A diagnosed of clear cell carcinoma of the right diaphragm without relation to endometriosis or ovarian malignancies was made. After resection of the tumor, it was revealed that her renal function was improvement. This is the first report of diaphragmatic clear cell carcinoma without relation to endometriosis or ovarian malignancies that caused secondary membranous nephropathy. One year after the resection of the tumor, she is being followed up and has shown no signs of recurrence.


Organochlorine pesticides in consumer fish and mollusks of Liaoning province, China: distribution and human exposure implications.
Fish and mollusk samples were collected from markets located in 12 cities in Liaoning province, China, during August and September 2007, and 22 organochlorine pesticides (OCPs) were detected. DDT, HCH, endosulfan, chlordane, and HCB were the dominating OCPs, with mean concentrations and ranges of, respectively, 15.41 and 0.57 to 177.56 ng/g, 0.84 and below detection limit (BDL) to 22.99 ng/g, 1.31 and BDL to 13.1 ng/g, 1.05 and BDL to 15.68 ng/g, and 0.63 and BDL to 9.21 ng/g in all fish and mollusk samples. The concentrations of other OCPs generally were low and were detectable in a minority of samples, reflecting the low levels of these OCPs in the study region. In general, OCP concentrations were obviously higher in fish than in mollusks, and higher in freshwater fish than in marine fish, which indicated, first, that freshwater fish are more easily influenced than seawater fish and mollusks by OCP residues in agricultural areas and, second, that there are different biota accumulation factors for OCPs between fish and mollusk. To learn the consumption of fish and mollusk, 256 questionnaires were sent to families in 12 cities of Liaoning province. Using the contamination data, average estimated daily intakes of OCPs via fish and mollusk consumption were calculated, which were used for exposure assessment. The public health risks caused by exposure to OCPs in the course of fish and mollusk consumption were compared to noncancer benchmarks and cancer benchmarks.


Moderate physical exercise improves lymphocyte function in melanoma-bearing mice on a high-fat diet.
Obesity can lead to a chronic systemic inflammatory state that increases the risk of cancer development. Therefore, this study aimed to evaluate the alterations in tumor non-infiltrated lymphocytes function and melanoma growth in animals maintained on a high-fat diet and/or moderate physical exercise program in a murine model of melanoma. Female mice were randomly divided into eight groups: 1) normolipidic control (N), 2) normolipidic + melanoma (NM), 3) high-fat control (H), 4) high-fat + melanoma (HM), 5) normolipidic control + physical exercise (NE), 6) normolipidic melanoma + physical exercise (NEM), 7) high-fat control + physical exercise (HE), and 8) high-fat melanoma + physical exercise (HEM). After 8 weeks of diet treatment and/or moderate physical exercise protocol, melanoma was initiated by explanting B16F10 cells into one-half of the animals. Animals fed a high-fat diet presented high-energy consumption (30%) and body weight gain (H and HE vs N and NE, 37%; HM and HEM vs NM and NEM, 73%, respectively), whether or not they carried melanoma explants. Although the tumor growth rate was higher in animals from the HM group than in animals from any other sedentary group, it was reduced by the addition of a physical exercise regimen. We also observed an increase in stimulated peripheral lymphocyte proliferation and a decrease in the T-helper 1 response in the HEM group. The results of the present study support the hypothesis that altering function of tumor non-infiltrated lymphocytes via exercise-related mechanisms can slow melanoma progression, indicating that the incorporation of a regular practice of moderate-intensity exercises can be a potential strategy for current therapeutic regimens in treating advanced melanoma.


Hepatitis C virus negative strand RNA is not detected in peripheral blood mononuclear cells and viral sequences are identical to those in serum: a case against extrahepatic replication.
Peripheral blood mononuclear cells (PBMCs) from 27 hepatitis C virus (HCV)-infected patients were analysed for the presence of HCV negative strand RNA with strand-specific Tth-based RT-PCR. No negative strand RNA was detected in any sample, and positive strand HCV sequences amplified from PBMCs were identical to those found in serum. These findings suggest that HCV does not replicate in PBMCs, and the presence of HCV sequences at this site is compatible with passive virus adsorption and/or contamination by circulating virus.


Effect of carbon dioxide on the formation of alpha-amylase by Bacillus subtilis growing in continuous and batch cultures.
The influence of CO2 on the information of alpha-amylase by Bacillus subtilis NCIB 8646 growing in continuous and batch cultures was investigated. Different levels of CO2 examined in the batch cultures stimulated the formation of alpha-amylase, with the highest activity being obtained using 6% CO2 (v/v). The additions of CO2 inhibit the growth and division of vegetative cells of B. subtilis when CO2 is present in a concentration of more than 3% (v/v). In chemostat cultures, air containing 8% CO2 (v/v) increased the specific enzyme productivity almost three times over the control, without affecting the cell growth. An attempt is made to correlate the obtained results with the alesis.


Predictors of mortality in beta-hemolytic streptococcal bacteremia: a population-based study.
Several factors associated with mortality in Lancefield group A beta-hemolytic streptococcal bacteremia have been described in population-based surveillance studies, whereas such reports on group B, C, and G streptococcal are scant. In this population-based study all 314 episodes of beta-hemolytic streptococcal bacteremia in adult patients in the Pirkanmaa area, Finland, during the 10-year period 1995-2004 were retrospectively reviewed. The 30-day case-fatality rate was 13%, being highest in group C (22%); in group A it was 15%, in group B 7%, and in group G 15%. Confusion, unconsciousness and dyspnea as the first sign or symptom were associated with increased case-fatality, while fever seemed to be a protecting factor for death. Alcoholism and ultimately or rapidly fatal underlying disease were significantly associated with increased case-fatality. Among infections of the skin and soft-tissues, necrotizing fasciitis had the highest risk of death (38%), while patients with cellulitis had a case-fatality of 8%. A history of previous cellulitis seemed to protect against death (case-fatality of 3% as compared to 16% among those without such a history (p=0.014)). A history of previous cellulitis seemed to be a protecting factor against death. Fever was also associated with a good prognosis.


Epidural epinephrine and the systemic circulation during peripheral vascular surgery.
This study was designed to determine the haemodynamic effects of epidural epinephrine, 5 micrograms.ml-1, added to bupivacaine, 0.75 per cent, in elderly patients with cardiac disease undergoing peripheral vascular surgery (PVS). The effect of epidural epinephrine on the plasma concentration of bupivacaine was also measured. Twenty patients with a history and/or ECG evidence of myocardial ischaemia requiring PVS were randomly assigned to two groups. The patients were monitored with a modified V5 ECG, oscillometric BP monitor and a PA catheter. After control haemodynamic measurements, 12 ml of bupivacaine, 0.75 per cent, +/- epinephrine, 5 micrograms.ml-1, was injected over five minutes into the epidural space at L3-4. Supine haemodynamic measurements were repeated at 15 and 45 min after injection. At 15 min after epidural injection, compared with control values, patients receiving epidural epinephrine showed a significantly greater decrease in mean blood pressure and systemic vascular resistance, and a significantly greater increase in cardiac output than patients receiving plain epidural bupivacaine (79.3 +/- 11.6 per cent vs 94.6 +/- 16.8 per cent, 61.6 +/- 9.0 vs 91.6 +/- 19.2 per cent, 130.8 +/- 23 vs 105 +/- 20.8 per cent, respectively). These differences were not present at 45 min after epidural injection. Heart rate was not significantly different between groups at either time. The presence of epidural epinephrine reduced the peak plasma concentration of bupivacaine from 0.86 +/- 0.20 to 0.64 +/- 0.33 micrograms.ml-1 and increased the time to achieve this concentration from 16.1 +/- 11.2 to 33.7 +/- 20.1 min.


Phialolunulospora vermispora (Chaetosphaeriaceae, Sordariomycetes), a novel asexual genus and species from freshwater in southern China.
The asexual taxon Phialolunulospora vermispora gen. et sp. nov., collected from submerged dicotyledonous leaves in Hainan, China, is described and illustrated herein. Phialolunulospora gen. nov. is characterized by macronematous, semimacronematous, septate and pigmented conidiophores and acrogenous, long lunate, vermiform to sigmoid, hyaline conidia with an eccentric basal appendage. Complete sequences of internal transcribed spacer (ITS) and partial sequences of nuclear large subunits ribosomal DNA (LSU) genes are provided. Phylogenetic analyses of combined ITS and LSU sequences revealed its placement in the Chaetosphaeriaceae. The new fungus is compared with morphologically similar genera.


Characterization of anti-thymoglobulin, anti-Atgam and anti-OKT3 IgG antibodies in human serum with an 11-min ELISA.
Patients treated with OKT3 may become resistant to OKT3 therapy following induction of anti-idiotypic antibodies. Antirabbit or antihorse antibodies following Thymoglobulin (rabbit antithymocyte globulin (ATG) or Atgam (horse ATG) therapy may not similarly inhibit drug efficacy due to an insufficient anti-idiotypic response against the multiple idiotypic specificities of the polyclonal anti-T cell preparations. However, no standardized assay had been developed to monitor antihorse and antirabbit antibodies. To address this issue, we developed three rapid (11-min) and standardized semiquantitative plate enzyme-linked immunoassays (ELISAs) to monitor human serum immunoglobulin-G (IgG) antibodies to Orthoclone OKT3 IgG2a, Atgam and Thymoglobulin. The format was identical for the three assays, with the exception of the immunoglobulin antigen coated on the ELISA plates. As OKT3 is a monoclonal antibody, OKT3 itself is used as the capture antigen. However, for antibody responses against the polyclonal antibody preparations Atgam and Thymoglobulin, it was found that horse IgG and rabbit IgG respectively were equivalent to Atgam and Thymoglobulin as capture antigens. Excellent correlation with a 3.5-h format was demonstrated (r values between 0.986 and 0.845). Specificity was demonstrated by inhibition experiments. Correlations between endpoint titres calculated from serial dilutions and 1:50 dilution OD values were 0.821, 0.983 and 0.937 respectively for the mouse, horse and rabbit assay. High titre sera were selected to assess their ability to block in vitro the binding of OKT3, Thymoglobulin and Atgam to T cells, using flow cytometry. None of the sera containing antihorse (n = 9) or antirabbit (n = 8) antibodies blocked T cell binding of Atgam or Thymoglobulin. In contrast, OKT3 binding was blocked by the four highest titre sera of the 13 anti-OKT3 sera tested. Consequently, prospective monitoring of treated patients using standardized 11-min assays may allow a better assessment of the effect of presensitization to OKT3, Atgam or Thymoglobulin.


Epidemiology of bacteremic urinary tract infections in chronically hospitalized elderly men.
We sought to determine the contribution of pathogen-specific factors to the pathogenesis of invasive urinary tract infections in chronically institutionalized elderly men with symptomatic bacteriuria. We found that Escherichia coli was the most invasive pathogen, being found in 46% of the bacteremic as opposed to 25% of the nonbacteremic, rigorously defined urinary tract infections (p less than 0.01). The predominance of E. coli in bacteremic urinary tract infections was observed regardless of whether indwelling urinary drainage devices were used. The finding that E. coli accounted for a greater proportion of bacteremic than nonbacteremic urinary tract infections indicates that bacteremia arising from the geriatric urinary tract is not simply a consequence of mechanical factors, such as urinary tract obstruction. Thus, further investigations are warranted to assess the specific contribution of previously defined E. coli virulence factors to the pathogenesis of urinary tract infections in such patients.


Mitochondrial enzyme GLUD2 plays a critical role in glioblastoma progression.
Glioblastoma (GBM) is the most frequent and malignant primary brain tumor in adults and despite the progress in surgical procedures and therapy options, the overall survival remains very poor. Glutamate and α-KG are fundamental elements necessary to support the growth and proliferation of GBM cells. Glutamate oxidative deamination, catalyzed by GLUD2, is the predominant pathway for the production of α-KG. GLUD2 emerged from the RNA-seq analysis of 13 GBM patients, performed in our laboratory and a microarray analysis of 77 high-grade gliomas available on the Geo database. Thereafter, we investigated GLUD2 relevance in cancer cell behavior by GLUD2 overexpression and silencing in two different human GBM cell lines. Finally, we overexpressed GLUD2 in-vivo by using zebrafish embryos and monitored the developing central nervous system. GLUD2 expression was found associated to the histopathological classification, prognosis and survival of GBM patients. Moreover, through in-vitro functional studies, we showed that differences in GLUD2 expression level affected cell proliferation, migration, invasion, colony formation abilities, cell cycle phases, mitochondrial function and ROS production. In support of these findings, we also demonstrated, with in-vivo studies, that GLUD2 overexpression affects glial cell proliferation without affecting neuronal development in zebrafish embryos. We concluded that GLUD2 overexpression inhibited GBM cell growth suggesting a novel potential drug target for control of GBM progression. The possibility to enhance GLUD2 activity in GBM could result in a blocked/reduced proliferation of GBM cells without affecting the survival of the surrounding neurons.


Price and reimbursement of advanced therapeutic medicinal products in Europe: are assessment and appraisal diverging from expert recommendations?
Advanced therapy medicinal products (ATMPs) represent an important cornerstone for innovation in healthcare. However, uncertainty on the value, the high average cost per patient and their one-shot nature has raised a debate on their assessment and appraisal process for pricing and reimbursement (P&R) purposes. This debate led experts providing for recommendations on this topic. Our primary objective is to investigate the ATMPs P&R process in the main five European countries and to understand if this process is consistent with published P&R expert recommendations. We also investigated the current ATMP pipelines to understand if future ATMPs will create challenges for their P&R process. P&R framework for ATMPs in the European Major five (EU5) countries was investigated through a literature search on PubMed, institutional websites of National Health Authorities and grey literature. The ATMPs pipeline database was populated from a clinical trial database (clinicaltrials.gov), relying on inclusion and exclusion criteria retrieved from the literature. Reimbursement status of ATMPs is different across the EU5 countries, with the exception of CAR-Ts which are reimbursed in all countries. Standard P&R process in place for other medicinal products is extended to ATMPs, with the exception of some cases in Germany. List prices, where available, are high and, tend to be aligned across countries. Outcome-based Managed Entry Agreements (MEAs) have been extensively used for ATMPs. Extra-funds for hospitals managing ATMPs were provided only in Germany and, as additional fund per episode, in France. The accreditation process of hospitals for ATMPs management was in most countries managed by the national authorities. As far as ATMPs pipeline is concerned, ATMPs in development are mostly targeting non-rare diseases. Expert recommendations for ATMPs P&R were partially applied: the role of outcome-based MEAs has increased and the selection process of the centres authorized to use these treatments has been enhanced; additional funding for ATMPs management to accredited centres has not been completely considered and annuity payment and broader perspective in cost considerations are far from being put in place. These recommendations should be considered for future P&R negotiations to pursue rational resource allocation and deal with budget constraints.


Recent trends in T7 phage application in diagnosis and treatment of various diseases.
The T7 phage is a virulent phage hosted by Escherichia coli, which poses no threat to animals and plants. Due to the advantages of small genome, well elucidated functional genomics, fast life cycle, and high stability, T7 phage has been widely used in many fields, including biology and medicine. In this review, we focus on the research of T7 phages in biological sciences and medicine, including the application of T7 phages and T7 phage products, T7 phage display systems, and recombinant T7 phages in the treatment and diagnosis of infectious diseases (bacteria, viruses, parasites) and tumor diseases. In addition, we also introduce the therapeutic application of T7 phage in various diseases such as allergic reaction, Alzheimer's disease, inflammatory reaction, and other diseases, and finally discuss the future direction of T7 phage application in the biomedical field.


Explorative analysis of a score predicting the therapy response of patients with metastatic, castration resistant prostate cancer undergoing radioligand therapy with 177Lu-labeled prostate-specific membrane antigen.
Up to 60% of patients with metastatic, castration-resistant prostate cancer (mCRPC) treated with 177Lu prostate-specific membrane antigen (PSMA) radioligand therapy (RLT) achieves a partial biochemical response with a decrease of > 50% in prostate-specific antigen (PSA) levels. The remaining fractions, however, do not respond to RLT. The aim of this explorative analysis was to identify pre-therapeutic factors for the prediction of response. 46 patients [age = 68 years (50-87)] with mCRPC who consecutively underwent RLT with 177Lu PSMA [median applied activity = 6 GBq (2.9-6.2)] were included and analysed retrospectively. The association of different clinical and laboratory factors and parameters from pre-therapeutic 68Ga PSMA positron emission tomography (PET) with the outcome of RLT was tested (Fisher's test). Outcome was defined as PSA changes 8 weeks after second RLT [partial response (PR), PSA decrease > 50%; progressive disease (PD), PSA increase ≥ 25%; stable disease (SD), others]. Significant predictive factors were combined in a predictive score. 30% showed a post-treatment PR (median 73% PSA decrease), 35% SD (median 17% PSA decrease) and 35% PD (median 42% PSA increase). Significant predictors for PD were alkaline phosphatase (ALP) > 135 U/l (p = 0.002), PSA > 200 ng/ml (p = 0.036), and maximum standardized uptake value (SUVmax) of the "hottest lesion" in pre-therapeutic PET < 45 (p = 0.005). The predictive score including PSA, ALP and SUVmax could separate 2 distinct groups of patients: ≤ 2 predictive factors (19% PD) and 3 predictive factors (90% PD). The presented predictive score allowed a pre-therapeutic estimate of the expected response to 2 cycles of RLT. As our study was retrospective, prospective trials are needed for validation.


Evaluation of repeatability and reproducibility of radiomic features produced by the fan-beam kV-CT on a novel ring gantry-based PET/CT linear accelerator.
The RefleXion X1 is a novel radiotherapy delivery system on a ring gantry equipped with fan-beam kV-CT and PET imaging subsystems. The day-to-day scanning variability of radiomics features must be evaluated before any attempt to utilize radiomics features. This study aims to characterize the repeatability and reproducibility of radiomic features produced by the RefleXion X1 kV-CT. The Credence Cartridge Radiomics (CCR) phantom includes six cartridges of varied materials. It was scanned 10 times on the RefleXion X1 kVCT imaging subsystem over a 3-month period using the two most frequently used scanning protocols (BMS and BMF). Fifty-five radiomic features were extracted for each ROI on each CT scan and analyzed using LifeX software. The coefficient of variation (COV) was computed to evaluate the repeatability. Intraclass correlation coefficient (ICC) and concordance correlation coefficient (CCC) were used to evaluate the repeatability and reproducibility of the scanned images using 0.9 as the threshold. This process is repeated on a GE PET-CT scanner using several built-in protocols as a comparison. On average, 87% of the features on both scan protocols on the RefleXion X1 kVCT imaging subsystem can be considered repeatable as they met COV < 10% criteria. On GE PET-CT, this number is similar at 86%. When we tighten the criteria to COV <5%, the RefleXion X1 kVCT imaging subsystem showed much better repeatability with 81% of features on average whereas GE PET-CT showed only 73.5% on average. About 91% and 89% of the features with ICC > 0.9 respectively for BMS and BMF protocols on RefleXion X1. On the other hand, the percentage of features with ICC > 0.9 on GE PET-CT ranges from 67% to 82%. The RefleXion X1 kVCT imaging subsystem showed excellent intra-scanner reproducibility between the scanning protocols much better than the GE PET CT scanner. For the inter-scanner reproducibility, the percentage of features with CCC > 0.9 ranged from 49% to 80%. between X1 and GE PET-CT scanning protocols. Clinically useful CT radiomic features produced by the RefleXion X1 kVCT imaging subsystem are reproducible and stable over time, demonstrating its utility as a quantitative imaging platform.


Rapid microbiological tests for bloodstream infections due to multidrug resistant Gram-negative bacteria: therapeutic implications.
Treating severe infections due to multidrug-resistant Gram-negative bacteria (MDR-GNB) is one of the most important challenges for clinicians worldwide, partly because resistance may remain unrecognized until identification of the causative agent and/or antimicrobial susceptibility testing (AST). Recently, some novel rapid test for identification and/or AST of MDR-GNB from positive blood cultures or the blood of patients with bloodstream infections (BSIs) have become available. The objective of this narrative review is to discuss the advantages and limitations of different rapid tests for identification and/or AST of MDR-GNB from positive blood cultures or the blood of patients with BSI, as well as the available evidence on their possible role to improve therapeutic decisions and antimicrobial stewardship. Inductive PubMed search for publications relevant to the topic. The present review is structured in the following way: (a) rapid tests on positive blood cultures; (b) rapid tests directly on whole blood; (c) therapeutic implications. Novel molecular and phenotypic rapid tests for identification and AST show the potential for favourably influencing patients' outcomes and results of antimicrobial stewardship interventions by reducing both the time to effective treatment and the misuse of antibiotics, although the interpretation about their impact on actual therapeutic decisions and patients' outcomes is still complex. Factors such as feasibility and personnel availability, as well as the detailed knowledge of the local microbiological epidemiology, need to be considered very carefully when implementing novel rapid tests in laboratory workflows and algorithms. Providing high-level, comparable evidence on the clinical impact of rapid identification and AST is becoming of paramount importance for MDR-GNB infections, since in the near future rapid identification of specific resistance mechanisms could be crucial for guiding rapid, effective, and targeted therapy against specific resistance mechanisms.


[The category gender and meeting standards for medical education as a result].
The Medical School Hanover, Institute of History, Ethics and Philosophy in Medicine, in co-operation with the University of Bristol, School for Policy Studies, carried out a project to examine the meaning of gender in medical education and to detect methods to incorporate the gender-perspective from public health research and gender studies as an interdisciplinary programme into medical education. The duration of the project was limited to two years, and funded by the EU from 2001 to 2003. The study clearly shows that the category gender is being very slowly introduced into medical education at medical faculties and medical universities in Europe. Questions are primary focussed on biological differences between the sexes of patients, whereas gender studies touch gender relationships in occupations, interactions and communication, different styles of leadership etc. The adaptation of this approach may lead to a new understanding of the doctor-patient-interaction as well as of the relationship between teachers and students.


Inguinal herniation of a transplant ureter: rare cause of obstructive uropathy.
We present a rare case of late renal allograft failure from ureteral obstruction resulting from inguinal herniation. A 72-year-old man presented with an elevated creatinine and hydroureteronephrosis of a transplanted kidney on ultrasound. Noncontrast computed tomography demonstrated an inguinal hernia containing ureter, and a nephrostomy tube was placed. The hernia and ureter were temporarily reduced during antegrade stent insertion. Creatinine normalized and we performed inguinal herniorrhaphy with polypropylene mesh. The ureter was not reimplanted. Renal function remained stable after nephrostomy tube removal. Simple herniorrhaphy without ureteral reimplantation may fix the case of ureteral obstruction from inguinal herniation.


Heteroclitic serological response in esophageal and prostate cancer patients after NY-ESO-1 protein vaccination.
NY-ESO-1 is a prototypic cancer/testis antigen. In a recent phase I clinical trial, we vaccinated 13 patients bearing NY-ESO-1-expressing tumors with a complex of cholesterol-bearing hydrophobized pullulan (CHP) and NY-ESO-1 protein (CHP-NY-ESO-1) and showed efficient induction of NY-ESO-1 antibody, and CD4 and CD8 T cell responses using peripheral blood from the patients. In our study, we analyzed heteroclitic serological responses in those patients after vaccination. Serological response against 11 tumor antigens including MAGE-A1, MAGE-A3, MAGE-A4, CT7/MAGEC1, CT10/MAGEC2, CT45, CT46/HORMAD1, SOX2, SSX2, XAGE1B and p53 was examined by enzyme-linked immunosorbent assay (ELISA) using sera from ten vaccinated patients. Expression of tumor antigens was determined by reverse transcription-polymerase chain reaction or immunohistochemistry. Eight of nine patients who showed antibody responses against NY-ESO-1 also showed an antibody response against at least 1 of these 11 tumor antigens after vaccination. In one patient, seven tumor antigens were recognized. Specificity analysis of the antibody response by ELISA using control recombinant proteins and synthetic peptides and by Western blot showed that the response was not against His6-tag and/or bacterial products included in a preparation of CHP-NY-ESO-1 used for vaccination. Thus, heteroclitic serological responses appear to be indicative of the overall immune response against the tumor, and their analysis could be useful for immune monitoring in cancer vaccine.


Recruitment of Cdc42 through the GAP domain of RLIP participates in remodeling of the actin cytoskeleton and is involved in Xenopus gastrulation.
The transduction pathways that branch out of fibroblast growth factor signaling are essential for the induction of the mesoderm and the specification of the vertebrate body plan. One of these pathways is thought to control remodeling of the actin cytoskeleton through the Ral binding protein (RLIP also known as RalBP1), an effector of the small G protein Ral. RLIP contains a region of homology with the GTPase-activating protein (GAP) domain involved in the regulation of GTPases of the Rho family. We demonstrate here that the GAP domain of RLIP is responsible for the stability of the actin cytoskeleton in Xenopus laevis embryos. We also demonstrate that the complete N-terminal domain of RLIP containing the mu2 binding domain (mu2BD) and the GAP domain induces disruption of the actin cytoskeleton when targeted to the plasma membrane. Neither domain, however, has any effect on the actin cytoskeleton when individually targeted to the plasma membrane. We also determined that Cdc42-GDP, but neither Rac-GDP nor Rho-GDP, rescues the effect of expression of the membrane-localized Xenopus RLIP on the actin cytoskeleton. We show that the GAP domain of RLIP interacts in vivo with Cdc42-GTP and Cdc42-GDP. Finally, a single mutation (K244A) in the GAP sequence prevented embryos from gastrulating. These results demonstrate that to participate in the control of the actin cytoskeleton, RLIP needs its complete N-terminal region coding for the mu2BD and the GAP domain. We suggest that RLIP, by coordinating two complementary mechanisms, the endocytosis of clathrin-coated pits and the remodeling of cortical actin, participates in the gastrulation process.


Cellular Biomarkers in Mytilus galloprovincialis L. (Mediterranean Mussels) from Izmir Bay (Turkey).
In this study, cellular biomarkers and heavy metal concentration in the Mytilus galloprovincialis L. (Mediterranean mussels) collected from eight sites of Izmir Bay (Turkey) were determined to reveal water pollution for the first time in these stations. Results show that heavy metals (As, Cu, Hg, Zn, Cd, Sn, Pb) have been specified in mussels' tissues collected from all stations. According to GST, SOD, CAT activities, and TBARS contents, mussels in the outer bay have exposed more oxidative stress than the ones in the inner bay. Digestive gland tissues of them were showed more inhibition at AChE levels than gills. Also, abnormal nucleus rates and micronucleus frequencies (MN) were found to be higher in the inner bay than in the outer bay. This study showed that heavy metal pollution in different levels is an environmental issue on the Izmir Bay. Especially the coastal regions of the bay have been extremely affected by anthropogenic effects due to growing population.


Replacement of Valproic Acid with New Anti-Seizure Medications in Idiopathic Generalized Epilepsy.
Background: Little is known regarding the non-inferiority of new anti-seizure medications (ASMs) in terms of replacing valproic acid (VPA) in patients with idiopathic generalized epilepsy (IGE). We hypothesized that replacement of VPA with new ASMs would offer non-inferior or better control of seizure frequency. The purpose of this study was to compare epileptic seizure frequency between the subset of patients with IGE who were on VPA and the subset of patients with IGE who replaced VPA with new ASMs. Methods: Patients with IGE who were on or had been on VPA between January 2016 and March 2022 were divided into a group that replaced VPA with new ASMs (VPA-replace group) and a group that remained on VPA (VPA-continue group). We then compared the groups in terms of seizure frequency and myoclonus. Results: Of the 606 patients on VPA between January 2016 and March 2022, 156 patients with IGE were enrolled to this study (VPA-replace group, n = 68; VPA-continue group, n = 88). The VPA-replace group included significantly more females than the VPA-continue group (p < 0.001). The VPA-replace group also showed significantly higher seizure frequency before replacement (p < 0.001), but not after replacement (p = 0.074). Patients on monotherapy displayed improved seizure frequency with new ASMs (p < 0.001). Among the new ASMs, perampanel (PER) significantly improved seizure frequency (p = 0.002). Forty-two patients in the VPA-replace group who had myoclonus achieved significant improvements (p < 0.001). Among these, patients on PER monotherapy (p < 0.001) or PER + lamotrigine (0.016) showed significantly improved myoclonus scale scores. Conclusions: This study shows the non-inferiority of new ASMs compared to VPA, with better seizure control using new ASMs in subsets of patients with IGE on monotherapy.


CETP inhibitors downregulate hepatic LDL receptor and PCSK9 expression in vitro and in vivo through a SREBP2 dependent mechanism.
CETP inhibitors block the transfer of cholesteryl ester from HDL-C to VLDL-C and LDL-C, thereby raising HDL-C and lowering LDL-C. In this study, we explored the effect of CETP inhibitors on hepatic LDL receptor (LDLR) and PCSK9 expression and further elucidated the underlying regulatory mechanism. We first examined the effect of anacetrapib (ANA) and dalcetrapib (DAL) on LDLR and PCSK9 expression in hepatic cells in vitro. ANA exhibited a dose-dependent inhibition on both LDLR and PCSK9 expression in CETP-positive HepG2 cells and human primary hepatocytes as well as CETP-negative mouse primary hepatocytes (MPH). Moreover, the induction of LDLR protein expression by rosuvastatin in MPH was blunted by cotreatment with ANA. In both HepG2 and MPH ANA treatment reduced the amount of mature form of SREBP2 (SREBP2-M). In vivo, oral administration of ANA to dyslipidemic C57BL/6J mice at a daily dose of 50 mg/kg for 1 week elevated serum total cholesterol by approximately 24.5% (p < 0.05%) and VLDL-C by 70% (p < 0.05%) with concomitant reductions of serum PCSK9 and liver LDLR/SREBP2-M protein. Finally, we examined the in vitro effect of two other strong CETP inhibitors evacetrapib and torcetrapib on LDLR/PCSK9 expression and observed a similar inhibitory effect as ANA in a concentration range of 1-10 μM. Our study revealed an unexpected off-target effect of CETP inhibitors that reduce the mature form of SREBP2, leading to attenuated transcription of hepatic LDLR and PCSK9. This negative regulation of SREBP pathway by ANA manifested in mice where CETP activity was absent and affected serum cholesterol metabolism.


Nitrate reductase activity and nitrite levels in the saliva of habitual users of various tobacco products.
Nitrite found in human saliva is the product of the microbial reduction of nitrate circulating through the salivary glands. Saliva samples were collected under controlled conditions from volunteers who were habitual users of different types of tobacco products (tobacco chewers, cigarette smokers, bidi smokers and masheri users) and from controls. The saliva samples were analysed for nitrite levels and for nitrate reductase activity spectrophotometrically. Samples were collected from two different areas, Bombay (urban) and Ghodegaon (rural). Salivary nitrite levels in the control groups ranged from 1.6 ppm in Ghodegaon to 11 ppm in Bombay. The nitrite levels of the masheri-using groups from both locations and of the tobacco-chewing group from Bombay were significantly higher than those of the controls. A number of volunteers showed undetectable levels of salivary nitrate reductase. Volunteers who chewed tobacco or used masheri had higher levels of nitrate reductase activity than the controls. However, there did not seem to be any clear correlation between nitrite levels and nitrate reductase activity in the saliva.


Molecular cloning and expression in Escherichia coli of an exo-levanase gene from the endophytic bacterium Gluconacetobacter diazotrophicus SRT4.
Gluconacetobacter diazotrophicus produces levan from sucrose by a secreted levansucrase (LsdA). A levanase-encoding gene ( lsdB), starting 51 bp downstream of the lsdA gene, was cloned from strain SRT4. The lsdB gene (1605 bp) encodes a protein (calculated molecular mass 58.4 kDa) containing a putative 36-amino-acid signal peptide at the N-terminus. The deduced amino acid sequence shares 34%, 33%, 32%, and 29% identities with levanases from Actinomyces naeslundii, Bacillus subtilis, Paenibacillus polymyxa, and Bacteroides fragilis, respectively. The lsdB expression in Escherichia coli under the control of the T7 RNA polymerase promoter resulted in an active enzyme which hydrolyzed levan, inulin, 1-kestose, raffinose, and sucrose, but not melezitose. Levanase activity was maximal at pH 6.0 and 30 degrees C, and it was not inhibited by the metal ion chelator EDTA or the denaturing agents dithiothreitol and beta-mercaptoethanol. The recombinant LsdB showed a fourfold higher rate of hydrolysis on levan compared to inulin, and the reaction on both substrates resulted in the successive liberation of the terminal fructosyl residues without formation of intermediate oligofructans, indicating a non-specific exo-levanase activity.


Leisure time physical activity, risk of depressive symptoms, and inflammatory mediators: the English Longitudinal Study of Ageing.
To examine if inflammatory markers (CRP, fibrinogen) might partly explain the association between physical activity (PA) and risk of depression. The English Longitudinal Study of Ageing, a prospective study of community dwelling older adults. 4323 men and women (aged 63.4+/-9.7 yrs) free from depression at baseline. Self reported leisure time PA levels and depressive symptoms (a score of > or = 4 using the 8-item CES-D scale) were assessed at baseline and 4 yrs follow up. The inflammatory markers, CRP and fibrinogen, were assessed at a 2 yrs intermediate time point between baseline and follow up. At follow up 8% of the sample reported depressive symptomatology. In comparison with participants reporting none or light PA, the odds of depressive symptomatology for those reporting moderate or vigorous PA were 0.71 (95% CI, 0.54-0.95) and 0.58 (0.41-0.81), respectively, after adjustments for baseline CES-D score, age, gender, social-occupational class, smoking, alcohol, and chronic illness. Each standard unit increase in log CRP was associated with higher odds of depressive symptomatology at follow up (1.32, 1.13-1.55) and CRP was inversely associated with physical activity. The association between PA and depressive symptomatology was not, however, substantially modified by further adjustment for CRP (odds for none vs. vigorous PA=0.60, 0.43-0.84). These data suggest that low grade systemic inflammation, as indexed by CRP, is a risk marker for depressive symptomatology, although this mechanism explains only a modest (approximately 5%) amount of the association between PA and risk of depression.


Supersensitivity of analgesic responses to alpha 2-adrenergic agonists in genetically hypertensive rats.
This study compared the responsivity of alpha 2-adrenoceptors to agonists in normotensive (WKY) and genetically hypertensive (SH) rats. Clonidine produced a greater degree of analgesia in SH as compared to WKY rats. This analgesia was antagonized by yohimbine. Neither naphazoline nor 4-hydroxy-clonidine produced analgesia in SH or WKY rats. Our results suggest that the analgesic effects of clonidine in SH rats are probably mediated by supersensitive central alpha 2-adrenoceptors.


Linkage scan for quantitative traits identifies new regions of interest for substance dependence in the Collaborative Study on the Genetics of Alcoholism (COGA) sample.
Dependence on alcohol and illicit drugs frequently co-occur. Results from a number of twin studies suggest that heritable influences on alcohol dependence and drug dependence may substantially overlap. Using large, genetically informative pedigrees from the Collaborative Study on the Genetics of Alcoholism (COGA), we performed quantitative linkage analyses using a panel of 1717 SNPs. Genome-wide linkage analyses were conducted for quantitative measures of DSM-IV alcohol dependence criteria, cannabis dependence criteria and dependence criteria across any illicit drug (including cannabis) individually and in combination as an average score across alcohol and illicit drug dependence criteria. For alcohol dependence, LOD scores exceeding 2.0 were noted on chromosome 1 (2.0 at 213 cM), 2 (3.4 at 234 cM) and 10 (3.7 at 60 cM). For cannabis dependence, a maximum LOD of 1.9 was noted at 95 cM on chromosome 14. For any illicit drug dependence, LODs of 2.0 and 2.4 were observed on chromosome 10 (116 cM) and 13 (64 cM) respectively. Finally, the combined alcohol and/or drug dependence symptoms yielded LODs >2.0 on chromosome 2 (3.2, 234 cM), 10 (2.4 and 2.6 at 60 cM and 116 cM) and 13 (2.1 at 64 cM). These regions may harbor genes that contribute to the biological basis of alcohol and drug dependence.


Downregulation of TLR4/MyD88/p38MAPK and JAK/STAT pathway in RAW 264.7 cells by Alpinia galanga reveals its beneficial effects in inflammation.
Alpinia galanga, commonly known as greater galangal or raasna, is widely used in Ayurveda against various inflammatory disorders. It is also known as Kulinjan, Aratha, Rasna or Sugandhamula. Some of the Ayurvedic preparations using the rhizome of Alpinia galanga are Rasnadi kashayam, Rasna panchakam, Rasnapthakam, and Rasnarendadi. The aromatic rhizome is the source of the drug greater galangal and it is also used as a spice in South and South East Asia. However, the molecular mechanism of action of A galanga against inflammation remains poorly understood. To elucidate the anti-inflammatory effect of hydroalcoholic extract of Alpinia galanga rhizome. The mechanism of the anti-inflammatory effect of hydroalcoholic extract of Alpinia galanga (AGE) was investigated by enzyme-linked immunosorbent assay (ELISA), Western blot, and immunofluorescence in LPS stimulated murine macrophage cell line (RAW 264.7). HPLC analysis was done to elucidate the rich polyphenolic nature of AGE. The study showed that pre-treatment with AGE downregulated the release of pro-inflammatory mediators (IL-6, TNF-α, NO, and ROS) and stimulated the release of anti-inflammatory mediator IL-10 in LPS stimulated RAW 264.7 cells. The vital enzymes of inflammation (iNOS, COX-2, and MMP-9) were also downregulated by pre-treatment with AGE. AGE targeted the upstream elements of the inflammatory cascade by blocking LPS induced activation of TLR4 and JAK/STAT pathway. The phosphorylation of downstream kinases was significantly affected. The inhibition of nuclear translocation of NFκB further confirmed the specific inhibition of the TLR4 pathway. Particularly AGE inhibited the phosphorylation of JNK, p38, IκBα, and STAT. HPLC analysis of the AGE showed the polyphenol-rich nature of the extract. The results from this study provide firm evidence that AGE exerts its anti-inflammatory effect via modulation of TLR4 and JAK/STAT pathway.


A sensitive technique to clone low abundance receptor transcripts from single microdissected tissue punches.
Tissue microdissection is a rapidly growing technique with wide applicability in the field of gene expression analysis as improved RNA extraction and reverse-transcription polymerase chain reaction (RT-PCR) techniques provide the sensitivity to amplify transcription products from increasingly small numbers of cells. In spite of these advances, isolation, cloning and regional localization of rare or low-abundance mRNA from very small tissue samples remain a difficult and challenging task, especially when high degenerate primers are to be used. We have addressed this problem using a combination of optimized techniques and purification steps added between individual reaction steps. The extreme sensitivity resulting from these modifications permits cloning of new members of a closely homologous gene family from only one microdissected tissue sample and widens the applicability of tissue microdissection. Using this protocol, nested degenerate PCR primers were designed to amplify members of the large and relatively homologous olfactory receptor (OR) gene family from RNA extracted from 125-microm diameter punches of tissue microdissected from 16-microm sections of the main olfactory bulb (MOB) of the mouse. Levels of OR mRNA in these punches are extremely low, due to the small volume of tissue and the low abundance of OR mRNA in MOB tissue. Several ORs were amplified, cloned and sequenced from a series of individual tissue punches, and in situ hybridization was used to verify the presence of mRNA corresponding to the cloned OR sequences in MOB sections.


Treatment access for opioid use disorder among women with medicaid in Florida.
Opioid use disorder (OUD) remains prevalent. Medications for OUD (MOUD) are standard care for pregnant and non-pregnant women. Previous research has identified barriers to MOUD for women with Medicaid but did not account for the type of MOUD (methadone vs. buprenorphine) or pregnancy status. We examined access to MOUD by treatment type for pregnant and non-pregnant women with Medicaid in Florida. A secondary analysis of Florida "secret-shopper" data was conducted. Calls were made to clinicians from the 2018 Substance Abuse and Mental Health Services Administration provider list by women posing as either a pregnant or non-pregnant woman with OUD and Medicaid. We examined 546 calls to buprenorphine-waivered providers (BWP) and 139 to opioid treatment programs (OTP). Counts and percentages were used to describe caller success by type of treatment and pregnancy status. Chi-square tests were used to identify statistical differences. Only 42 % of calls reached a treatment provider in Florida. Pregnant and non-pregnant women were less likely to obtain an appointment with Medicaid coverage by a BWP than an OTP (p < 0.01). Nearly 40 % of OTPs offered appointments to callers with Medicaid compared to only 17 % of BWPs. Both types of providers denied appointments more often for pregnant women. Thirty-eight percent of BWP's and 12 % of OTP's denied appointments to pregnant women using cash or Medicaid payment. Our study demonstrates logistical and financial barriers to treatment for OUD among pregnant and non-pregnant women with Medicaid in Florida and highlights the need for improved systems of care.


Comparison of the major cell populations among osteoarthritis, Kashin-Beck disease and healthy chondrocytes by single-cell RNA-seq analysis.
Chondrocytes are the key target cells of the cartilage degeneration that occurs in Kashin-Beck disease (KBD) and osteoarthritis (OA). However, the heterogeneity of articular cartilage cell types present in KBD and OA patients and healthy controls is still unknown, which has prevented the study of the pathophysiology of the mechanisms underlying the roles of different populations of chondrocytes in the processes leading to KBD and OA. Here, we aimed to identify the transcriptional programmes and all major cell populations in patients with KBD, patients with OA and healthy controls to identify the markers that discriminate among chondrocytes in these three groups. Single-cell RNA sequencing was performed to identify chondrocyte populations and their gene signatures in KBD, OA and healthy cells to investigate their differences as related to the pathogenetic mechanisms of these two osteochondral diseases. We performed immunohistochemistry and quantitative reverse-transcription PCR (qRT-PCR) assays to validate the markers for chondrocyte population. Ten clusters were labelled by cell type according to the expression of previously described markers, and one novel population was identified according to the expression of a new set of markers. The homeostatic and mitochondrial chondrocyte populations, which were identified by the expression of the unknown markers MT1X and MT2A and MT-ND1 and MT-ATP6, were markedly expanded in KBD. The regulatory chondrocyte population, identified by the expression of CHI3L1, was markedly expanded in OA. Our study allows us to better understand the heterogeneity of chondrocytes in KBD and OA and provides new evidence of differences in the pathogenetic mechanisms between these two diseases.


Assessment of follicular maturity by follicular diameter and fluid volume in a program of in vitro fertilization and embryo transfer.
We compared the assessment of follicular maturity by follicular diameter or follicular fluid (FF) volume and the subsequent fertilization and cleavage of the oocytes in 36 patients who had ovarian stimulation performed for in vitro fertilization and embryo transfer (IVF-ET). All patients received clomiphene citrate and human menopausal gonadotropin. The daily growth of follicles was assessed ultrasonically for 6 consecutive days before oocyte collection. The follicular diameter was found to be well correlated with the volume of follicular fluid. There were no significant differences among the dominant, second, and third follicles in the rates of oocyte recovery, maturation, fertilization, or cleavage. The rates of oocyte maturation and fertilization in FF volumes of 5.1 to 7.0 ml were significantly higher than in volumes of 1.1 to 5.0 ml, but the cleavage rates were similar. A smaller percentage of embryos developed to the four-cell stage when the oocytes were recovered from follicles with volumes of 1.1 to 3.0 ml. These observations suggest that follicular volume is a good parameter for judging the maturity of oocytes. It is also suggested that, although the oocytes of small follicles can be fertilized and cleaved, oocytes from larger follicles may have a better developmental capacity.


Synaptophysin as a probable component of neurotransmission occurring in taste receptor cells.
Taste signal is received in taste buds and transmitted via sensory afferent nerves to the brainstem. Although a signaling pathway involving phospholipase C-beta2 has been shown to transduce taste signals of bitterness, sweetness and umami in taste receptor cells (Type II cells), these taste receptor cells appear to be different from the presynaptic cells (Type III cells) containing afferent synapses associated with nerve processes. To elucidate the neurotransmission system in the taste receptor cells expressing phospholipase C-beta2, we searched for candidate molecules involved in the neurotransmission, and identified synaptophysin. Synaptophysin was expressed in the taste receptor cells expressing phospholipase C-beta2, as well as in the presynaptic cells harboring synaptic structures with taste nerves and containing serotonin. Synaptophysin-immunoreactive signals were not limited to gustducin-positive bitter taste receptor cells, and sweet/umami taste receptor cells were indicated to also express synaptophysin. Expression of synaptophysin was already initiated 6 days after cell division, almost in synchrony with the initiation of phospholipase C-beta2 expression. Synaptophysin-containing cells co-expressed vesicular-associated membrane protein 2, a v-SNARE molecule which is important for exocytosis. In addition, majority of the synaptophysin-expressing cells also expressed cholecystokinin, a neuropeptide expressed in taste buds. These results suggest that the taste receptor cells have a neurotransmission system involving synaptophysin, which occurs alternatively or additionally to a recently shown hemichannel system.


[Results of the treatment of breast cancer (based on the materials of the oncological ambulatory services of the Dagestan Republic, 1969-1983)].
The paper discusses the results of a retrospective analysis of the effectiveness of breast cancer treatment in the Daghestan ASSR. The study was based on the data of medical histories of 1120 patients treated at the Dispensary within 1969-1983.


Coordinate value of the femoral head center estimated using those of the tip of the greater trochanter and lesser trochanter.
Several studies have reported estimating the femoral head center (FC) from reference points on the pelvis; however, none have reported estimates obtained from those on the femur. In this cross-sectional study, we investigated the estimated point of FC from the coordinate value of the tip of the greater trochanter (GT) and lesser trochanter (LT) using a formula with a three-dimensional measurement technique. We used data from 92 healthy Japanese subjects without any back or knee symptoms and no abnormalities in the hip, knee, or spine on plain radiographs. In our study, the difference in the anteroposterior direction was larger than that in the other directions. We speculate that the accuracy of defining the tip of the LT is difficult in the anteroposterior direction. Moreover, the correlation coefficients were larger for women. The reason for this was unclear because the variation in the proximal femur may be similar in women. We found that the average difference between the actual and calculated values was approximately 2 mm. We considered that the coordinate value of the FC from the tip of the GT could be estimated more accurately using the regression equation compared to previous methods based on pelvic reference points.


Synthesis and antibody recognition of cyclic epitope peptides, together with their dimer and conjugated derivatives based on residues 9-22 of herpes simplex virus type 1 glycoprotein D.
The synthesis of new cyclic peptides comprising the 9-22 epitope (9)LKMADPNRFRGKDL(22) sequence derived from HSV gD-1 is reported. In addition, we describe procedures for the preparation of cyclic peptide dimers and conjugates with an oligotuftsin derivative carrier. The binding of a monoclonal antibody, Mab A16, to the synthesized compounds was determined by enzyme-linked immunosorbent assay. It was demonstrated that cyclization decreased the binding activity of the antibody to the epitope. However, dimerization and conjugation could significantly increase the binding capacity of the cyclic epitope peptides. The attachment site in dimers and conjugates, as well as the topology of the construct, had a significant influence on the antibody recognition, while replacement of Met in position 11 by Nle had no marked effect.


High Clinical Burden of Influenza Disease in Adults Aged ≥ 65 Years: Can We Do Better? A Systematic Literature Review.
Influenza is a respiratory infection associated with a significant clinical burden globally. Adults aged ≥ 65 years are at increased risk of severe influenza-related symptoms and complications due to chronic comorbidity and immunosenescence. Annual influenza vaccination is recommended; however, current influenza vaccines confer suboptimal protection, in part due to antigen mismatch and poor durability. This systematic literature review characterizes the global clinical burden of seasonal influenza among adults aged ≥ 65 years. An electronic database search was conducted and supplemented with a conference abstract search. Included studies described clinical outcomes in the ≥ 65 years population across several global regions and were published in English between January 1, 2012 and February 9, 2022. Ninety-nine publications were included (accounting for > 156,198,287 total participants globally). Clinical burden was evident across regions, with most studies conducted in the USA and Europe. Risk of influenza-associated hospitalization increased with age, particularly in those aged ≥ 65 years living in long-term care facilities, with underlying comorbidities, and infected with A(H3N2) strains. Seasons dominated by circulating A(H3N2) strains saw increased risk of influenza-associated hospitalization, intensive care unit admission, and mortality within the ≥ 65 years population. Seasonal differences in clinical burden were linked to differences in circulating strains. Influenza exerts a considerable burden on adults aged ≥ 65 years and healthcare systems, with high incidence of hospitalization and mortality. Substantial influenza-associated clinical burden persists despite increasing vaccination coverage among adults aged ≥ 65 years across regions included in this review, which suggests limited effectiveness of currently available seasonal influenza vaccines. To reduce influenza-associated clinical burden, influenza vaccine effectiveness must be improved. Next generation vaccine production using mRNA technology has demonstrated high effectiveness against another respiratory virus-SARS-CoV-2-and may overcome the practical limitations associated with traditional influenza vaccine production.


Clinical Characteristics of Patients With Kawasaki Disease Whose Siblings Had the Same Disease.
Some patients with Kawasaki disease (KD) have siblings who developed the same disease. Using a large-scale epidemiologic dataset, the present study aimed to determine the clinical characteristics of this population. We analyzed 89,725 patients diagnosed with KD during 2011-2018 who were registered in the nationwide Japanese KD survey database. Multivariable logistic regression analyses were performed to determine factors associated with sibling history of KD. Of the 89,725 patients, 1777 (2%) had sibling history of KD. Annual prevalence ranged from 1.5% to 2.3% during the study period and showed a tendency toward an increasing trend. Patients with recurrent KD and parental history of KD were significantly associated with sibling history of KD (adjusted odds ratio [95% confidence interval] = 2.15 [1.82-2.54] and 2.64 [2.02-3.47], respectively). Although patients with a sibling history of KD were significantly associated with initial intravenous immunoglobulin treatment resistance (1.14 [1.02-1.28]), no significant association was found between sibling history and coronary artery abnormality development. Among patients with a sibling history of KD, male patients were less likely to have recurrent KD than female patients (0.68 [0.49-0.96]). The significant association between sibling history and parental history may indicate genetic susceptibility to KD onset. Among those with a sibling history, recurrent KD was more likely to occur in female patients. Further studies focusing on this population may contribute toward identification of the cause of KD onset.


Analysis of mechanical strength to fixing the femoral neck fracture in synthetic bone type Asnis.
To analyze the results of biomechanical assays of fixation of Pauwels type III femoral neck fracture in synthetic bone, using 7.5mm cannulated screws in inverted triangle formation, in relation to the control group. TEN SYNTHETIC BONES WERE USED, FROM A DOMESTIC BRAND, DIVIDED INTO TWO GROUPS: test and control. In the test group, a 70° tilt osteotomy of the femoral neck was fixated using three cannulated screws in inverted triangle formation. The resistance of this fixation and its rotational deviation were analyzed at 5mm displacement (phase 1) and 10mm displacement (phase 2). The control group was tested in its integrity until the fracture of the femoral neck occurred. The Mann-Whitney test was used for group analysis and comparison. The values in the test group in phase 1, in samples 1-5, showed a mean of 579N and SD =77N. Rotational deviations showed a mean of 3.33°, SD = 2.63°. In phase 2, the mean was 696N and SD =106N. The values of the maximum load in the control group had a mean of 1329N and SD=177N. The analysis of mechanical strength between the groups determined a statistically significant lower value in the test group. Level of Evidence III, Control Case.


Visual attention: the past 25 years.
This review focuses on covert attention and how it alters early vision. I explain why attention is considered a selective process, the constructs of covert attention, spatial endogenous and exogenous attention, and feature-based attention. I explain how in the last 25 years research on attention has characterized the effects of covert attention on spatial filters and how attention influences the selection of stimuli of interest. This review includes the effects of spatial attention on discriminability and appearance in tasks mediated by contrast sensitivity and spatial resolution; the effects of feature-based attention on basic visual processes, and a comparison of the effects of spatial and feature-based attention. The emphasis of this review is on psychophysical studies, but relevant electrophysiological and neuroimaging studies and models regarding how and where neuronal responses are modulated are also discussed.


Effect of Training Load Distribution on Aerobic Fitness Measures in Hurling Players.
Malone, S, Hughes, B, and Collins, K. Effect of training load distribution on aerobic fitness measures in hurling players. J Strength Cond Res 33(3): 825-830, 2019-The aim of the current investigation was to quantify the training intensity distribution of hurling players and their association with changes in aerobic fitness measures within hurling players. Thirty hurling players (n = 30; age 26.5 ± 3.2 years; height 178.9 ± 3.2 cm; body mass 81.5 ± 4.5 kg) were observed during the pre-championship training period (12 weeks). Aerobic fitness measures (V[Combining Dot Above]O2max), peak treadmill velocity (km·h), heart rates (HRs) at 2 and 4 mmol·L blood lactate concentrations, and Yo-Yo intermittent recovery test performance (Yo-YoIR1) were assessed before and after the training period. Training intensities were categorized using 3 individualized HR zones based on the following criteria: low intensity (≤HR 2 mmol·L), moderate intensity (between HR 2 and 4 mmol·L), and high intensity (≥HR 4 mmol·L). Analysis of 1,025 individual training sessions showed that players spent 58.1 ± 2.5%, 24.3 ± 2.9%, and 17.6 ± 1.3% of time in the low-, moderate-, and high-intensity zones, respectively. The training time spent at high intensity (Zone 3) showed moderate-to-large associations with improvements in S2 (r = 0.80; p = 0.04) and S4 (r = 0.58; p = 0.03). Similar trends were observed for time spent in high intensity and improvements in V[Combining Dot Above]O2max (r = 0.77; p = 0.001) and Yo-YoIR1 performance (r = 0.65; p = 0.05). Hurling players have been shown to spend the majority of training time in low (≤ HR 2 mmol·L) to moderate intensity (between HR 2 and 4 mmol·L) training. However, only the time spent at high intensity (≥90% of maximal HR) was related to changes in aerobic fitness. These results support the usefulness of the quantification of aerobic training load using HR. The study also supports the efficacy of spending elements of training time within the high-intensity zone to increase hurling player's aerobic fitness characteristics.


Mediation of the microbiome-gut axis by oyster (Crassostrea gigas) polysaccharides: A possible protective role in alcoholic liver injury.
The objective of this study was to evaluate the molecular mechanism by which polysaccharides from Crassostrea gigas (RPS) prevent alcoholic liver injury and to uncover whether the steaming process affects the bioactivities of RPS. Oral administration of RPS or polysaccharides from steamed oyster (SPS) (282 mg/kg b.w.) significantly attenuated alcoholic liver injury in mice. RPS and SPS treatments protected gut functions by significantly enhancing the expression of tight-junction proteins and suppressing inflammatory responses. RPS and SPS treatments also significantly increased Lactobacillus reuteri and Roseburia spp. and decreased the level of Escherichia. Microbial metabolites, especially propionate and butyrate, were also increased in RPS- and SPS-treated mice. Correlation analysis revealed that the beneficial effects of RPS and SPS were strongly correlated with the microbiota composition and SCFAs. These results indicated that oyster polysaccharides alleviated alcoholic liver injury by mediating the gut-liver-metabolite axis, and the steaming process had little influence on the bioactivity.


[Biomechanics and movements of the body after 120-day antiorthostatic hypokinesia].
This paper presents data on body parameters (weight and circumference) and walking biomechanics after 120-day head-down tilt. The exposure leads to changes primarily in the anti-gravitational muscles. They are assumed to be caused by relative changes in the fat and muscle components. Head-down tilt also produces changes in the kinematic parameters of the walking process, the shape of support reactions, and losses in the cost-efficiency of the walking process.


Preclinical cancer of the cervix: diagnostic pitfalls.
This paper outlines the pitfalls encountered in the diagnosis of 127 preclinical cancers of the cervix. Each patient had cervical cytology, colposcopy, and histology performed within the same institution. Early stromal invasion was diagnosed in 55 patients and occult invasion was diagnosed in 72 patients. For preclinical cancers as a group, the possibility of invasive carcinoma was predicted cytologically in 36% and suspected colposcopically in 41%. However, in over 85% of patients the abnormal transformation zone (TZ) extended into or was totally confined within the endocervical canal and could not be adequately evaluated with the colposcope. A feature of this study was the high incidence of incomplete excision of abnormal tissue by cone biopsy and a high incidence of residual disease at hysterectomy. Cone biopsy was necessary to accurately assess the maximum depth and extent of invasion prior to definitive therapy whenever invasion was suspected colposcopically or on target biopsy. In 10 of 16 such patients, the cone biopsy demonstrated occult invasion. Colposcopy and directed target biopsy alone was adequate for the diagnosis of frank invasive cancer in 22 of the 72 patients in the occult invasion group, enabling definitive therapy to be undertaken without further delay.


A Plasmodium falciparum lysophospholipase regulates host fatty acid flux via parasite lipid storage to enable controlled asexual schizogony.
Phospholipid metabolism is crucial for membrane biogenesis and homeostasis of Plasmodium falciparum. To generate such phospholipids, the parasite extensively scavenges, recycles, and reassembles host lipids. P. falciparum possesses an unusually large number of lysophospholipases, whose roles and importance remain to be elucidated. Here, we functionally characterize one P. falciparum lysophospholipase, PfLPL3, to reveal its key role in parasite propagation during asexual blood stages. PfLPL3 displays a dynamic localization throughout asexual stages, mainly localizing in the host-parasite interface. Inducible knockdown of PfLPL3 disrupts parasite development from trophozoites to schizont, inducing a drastic reduction in merozoite progenies. Detailed lipidomic analyses show that PfLPL3 generates fatty acids from scavenged host lipids to generate neutral lipids. These are then timely mobilized to allow schizogony and merozoite formation. We then identify inhibitors of PfLPL3 from Medicine for Malaria Venture (MMV) with potent antimalarial activity, which could also serve as pertinent chemical tools to study parasite lipid synthesis.


Complete genome sequence of the halophile bacterium Kushneria konosiri X49T, isolated from salt-fermented Konosirus punctatus.
Kushneria konosiri X49T is a member of the Halomonadaceae family within the order Oceanospirillales and can be isolated from salt-fermented larval gizzard shad. The genome of K. konosiri X49T reported here provides a genetic basis for its halophilic character. Diverse genes were involved in salt-in and -out strategies enabling adaptation of X49T to hypersaline environments. Due to resistance to high salt concentrations, genome research of K. konosiri X49T will contribute to the improvement of environmental and biotechnological usage by enhancing understanding of the osmotic equilibrium in the cytoplasm. Its genome consists of 3,584,631 bp, with an average G + C content of 59.1%, and 3261 coding sequences, 12 rRNAs, 66 tRNAs, and 8 miscRNAs.


N-Acetylcysteine treatment of rotavirus-associated diarrhea in children.
Rotaviruses are the leading cause of severe, acute, and dehydrating diarrhea affecting children under 5 years of age worldwide. Despite an important reduction in rotavirus-caused deaths as a consequence of the rotavirus vaccine, alternative or complementary strategies for preventing or treating rotavirus-associated diarrhea are needed mainly in the poorest countries. We describe the cases of four rotavirus-unvaccinated 12-13-month-old girls and a 5-year-old boy who developed rotavirus-associated diarrhea confirmed by enzyme-linked immunosorbent assay, Western blotting, and immunochemistry analyses. After the first day of diarrheal episodes, three of the five patients were immediately administered oral N-acetylcysteine (NAC) 60 mg/kg daily, divided into three equal doses every 8 hours. The other two patients did not receive NAC and served as controls. Administration of NAC resulted in a decreased number of diarrheal episodes, excretion of fecal rotavirus antigen, and resolution of symptoms after 2 days of treatment. Our results suggest that NAC treatment after the first diarrheal episode could be an efficient strategy for treating rotavirus-affected children and preventing the associated severe life-threatening accompanying dehydration.


3-[2-(6-Bromo-2-phenyl-3H-imidazo[4,5-b]pyridin-3-yl)eth-yl]-1,3-oxazolidin-2-one.
In the title mol-ecule, C(17)H(15)BrN(4)O(2), the fused-ring system is essentially planar, the largest deviation from the mean plane being 0.015 (2) Å, and forms dihedral angles of 37.8 (2) and 35.5 (2)° with the phenyl and oxazolidine rings, respectively. The conformation adopted by the mol-ecule is stabilized by an intra-molecular π⋯π inter-action [centroid-centroid distance = 3.855(2) Å] between oxazolidine and phenyl rings. The crystal packing features inter-molecular C-H⋯N and C-H⋯O inter-actions.


How does portfolio use affect self-regulated learning in clinical workplace learning: What works, for whom, and in what contexts?
Portfolio use to support self-regulated learning (SRL) during clinical workplace learning is widespread, but much is still unknown regarding its effectiveness. This review aimed to gain insight in the extent to which portfolio use supports SRL and under what circumstances. A realist review was conducted in two phases. First, stakeholder interviews and a scoping search were used to formulate a program theory that explains how portfolio use could support SRL. Second, an in-depth literature search was conducted. The included papers were coded to extract context-mechanism-outcome configurations (CMOs). These were synthesized to answer the research question. Sixteen papers were included (four fulfilled all qualitative rigor criteria). Two primary portfolio mechanisms were established: documenting as a moment of contemplation (learners analyze experiences while writing portfolio reports) and documentation as a reminder of past events (previous portfolio reports aid recall). These mechanisms may explain the positive relationship between portfolio use and self-assessment, reflection, and feedback. However, other SRL outcomes were only supported to a limited extent: formulation of learning objectives and plans, and monitoring. The partial support of the program theory can be explained by interference of contextual factors (e.g., system of assessment) and portfolio-related mechanisms (e.g., mentoring). Portfolio research is falling short both theoretically-in defining and conceptualizing SRL-and methodologically. Nevertheless, this review indicates that portfolio use has potential to support SRL. However, the working mechanisms of portfolio use are easily disrupted. These disruptions seem to relate to tensions between different portfolio purposes, which may undermine learners' motivation.


Survival in atrioventricular discordance.
Limited information is available concerning the long-term survival of patients with atrioventricular discordance, ventriculoarterial discordance, and two ventricles (corrected transposition). The long-term follow-up of 107 patients examined at the Mayo Clinic over a 30-year period between 1951 and 1981 was reviewed. Overall survival from the date of Mayo Clinic diagnosis was 70% at five years and 64% at ten years. Associated variables were analyzed for their effect on survival including sex, age at diagnosis, presence of ventricular septal defect, pulmonary stenosis, dextrocardia, left atrioventricular valve insufficiency, and complete heart block. There was no significant difference between those with and without a ventricular septal defect (VSD). Pulmonary stenosis was protective when a VSD was present but was not a significant predictor of long-term survival. The only variable that consistently correlated with decreased survival was left atrioventricular valve insufficiency (p less than 0.04 for univariate and stepwise Cox, and p = 0.08 for multivariate analysis). A logistic model for survival after open-heart surgery failed to identify any significant variable. We conclude that the presence of left atrioventricular valve insufficiency in association with atrioventricular discordance significantly alters the long-term outcome. Atrioventricular valve replacement should be considered in such patients when insufficiency becomes hemodynamically significant.


Interplay between the prostaglandin transporter OATP2A1 and prostaglandin E2-mediated cellular effects.
Prostaglandins such as prostaglandin E2 (PGE2) play a pivotal role in physiological and pathophysiological pathways in gastric mucosa. Little is known about the interrelation of the prostaglandin E (EP) receptors with the prostaglandin transporter OATP2A1 in the gastric mucosa and gastric carcinoma. Therefore, we first investigated the expression of OATP2A1 and EP4 in normal and carcinoma gastric mucosa. Different PGE2-mediated cellular pathways and mechanisms were investigated using human embryonic kidney cells (HEK293) and the human gastric carcinoma cell line AGS stably transfected with OATP2A1. Colocalization and expression of OATP2A1 and EP4 were detected in mucosa of normal gastric tissue and of gastric carcinomas. OATP2A1 reduced the PGE2-mediated cAMP production in HEK293 and AGS cells overexpressing EP4 and OATP2A1. The expression of OATP2A1 in AGS cells resulted in a reduction of [(3)H]-thymidine incorporation which was in line with a higher accumulation of AGS-OATP2A1 cells in S-phase of the cell cycle compared to control cells. In contrast, the expression of OATP2A1 in HEK293 cells had no influence on the distribution in the S-phase compared to control cells. OATP2A1 also diminished the PGE2-mediated expression of interleukin-8 mRNA (IL-8) and hypoxia-inducible-factor 1α (HIF1α) protein in AGS-OATP2A1 cells. The expression of OATP2A1 increased the sensitivity of AGS cells against irinotecan which led to reduced cell viability. Taken together, these data show that OATP2A1 influences PGE2-mediated cellular pathways. Therefore, OATP2A1 needs to be considered as a key determinant for the understanding of the physiology and pathophysiology of prostaglandins in healthy and tumorous gastric mucosa.


Sarcopenia Prevalence Using Different Definitions in Older Community-Dwelling Canadians.
Sarcopenia is a debilitating condition affecting millions of individuals worldwide and is defined with different criteria. The objective of this study was to determine the prevalence of sarcopenia in older Canadians using three internationally accepted criteria. Observational cohort study. Data from 12,592 subjects [6,314 males (50.1%), 6,278 females (49.9%)] ≥65 years old in the Canadian Longitudinal Study on Aging were included. Appendicular lean mass (ALM; kg) and appendicular lean mass index (ALM kg/height in m2) were collected from dual X-ray absorptiometry measurements. Physical performance was assessed using the 4-m gait speed test, and muscle strength was measured by hand dynamometry. Sarcopenia was defined according to criteria put forth by the International Working Group on Sarcopenia (IWGS), Foundation for the National Institutes of Health (FNIH) Sarcopenia Project, and revised European Working Group on Sarcopenia in Older People (EWGSOP). Positive and negative percent agreements and Cohen's kappa (κ) described the agreement among sarcopenia definitions. Among the evaluated criteria, gait speed ≤ 1.0 m/s (IWGS definition of slowness) was the most frequently identified deficit (56.8% males, 57.2% females). The prevalence of sarcopenia ranged from 1.4 to 5.2% in males and 1.6 to 7.2 % in females among the different definitions. Positive percent agreement values among criteria were generally low (range: 1.5 - 55.3%) and corresponded to κ indicating none to minimal agreement (0.01 - 0.23). Negative percent agreement values were ≥ 95%. Sarcopenia prevalence was relatively low in older Canadian adults and current definitions had poor agreement in diagnosing individuals as sarcopenic.


Become the PPUPET Master: Mastering Pressure Ulcer Risk Assessment With the Pediatric Pressure Ulcer Prediction and Evaluation Tool (PPUPET).
Hospital acquired pressure ulcers (HAPU) are serious, debilitating, and preventable complications in all inpatient populations. Despite evidence of the development of pressure ulcers in the pediatric population, minimal research has been done. Based on observations gathered during quarterly HAPU audits, bedside nursing staff recognized trends in pressure ulcer locations that were not captured using current pressure ulcer risk assessment tools. Together, bedside nurses and nursing leadership created and conducted multiple research studies to investigate the validity and reliability of the Pediatric Pressure Ulcer Prediction and Evaluation Tool (PPUPET).


The high performance parameterization for deep learning in pulse shaping.
In high energy physics, front-end data acquisition systems based on analog-to-digital converters (ADC) can provide multiple aspects (time, energy, position) of information when an incident particle is detected. To process the shaped semi-Gaussian pulses from ADCs, multi-layer neural networks (aka. deep learning recently) show excellent accuracy and promising real-time capability. However, several factors, such as sampling rate and precision, neural network quantization bits, and intrinsic noise, complicate the problem and make it hard to find a cost-effective solution with high performance. In this article, we analyze above factors in a systematic way to study the effect of each one on the performance of the network individually when other factors are controlled. Moreover, the proposed network architecture can provide both the time and energy information from a single pulse. When the sampling rate is 2.5 MHz, sampling precision is 5-bit, the network tested in this work with an 8-bit encoder and a 16-bit decoder (designated as N2) achieved the best comprehensive performance in all conditions.


Flattening of the distal femoral epiphyses in the trichorhinophalangeal syndrome.
A new finding in the trichorhinophalangeal syndrome, flattening of the distal femoral epiphyses, is described in seven patients. Measurement of the ratio of the height of the distal femoral epiphysis versus both the width of the distal femoral metaphysis and the width of the distal femoral epiphysis and comparison to normal standards confirmed the subjectively observed flattening of the distal femoral epiphysis in these patients, further suggesting that the disease is a generalized process rather than a local disorder.


HIF-1 and NRF2; Key Molecules for Malignant Phenotypes of Pancreatic Cancer.
Pancreatic cancer is intractable due to early progression and resistance to conventional therapy. Dense fibrotic stroma, known as desmoplasia, is a characteristic feature of pancreatic cancer, and develops through the interactions between pancreatic cancer cells and stromal cells, including pancreatic stellate cells. Dense stroma forms harsh tumor microenvironments characterized by hypoxia, few nutrients, and oxidative stress. Pancreatic cancer cells as well as pancreatic stellate cells survive in the harsh microenvironments through the altered expression of signaling molecules, transporters, and metabolic enzymes governed by various stress response mechanisms. Hypoxia inducible factor-1 and KEAP1-NRF2, stress response mechanisms for hypoxia and oxidative stress, respectively, contribute to the aggressive behaviors of pancreatic cancer. These key molecules for stress response mechanisms are activated, both in pancreatic cancer cells and in pancreatic stellate cells. Both factors are involved in the mutual activation of cancer cells and stellate cells, by inducing cancer-promoting signals and their mediators. Therapeutic interventions targeting these pathways are promising approaches for novel therapies. In this review, we summarize the roles of stress response mechanisms, focusing on hypoxia inducible factor-1 and KEAP1-NRF2, in pancreatic cancer. In addition, we discuss the potential of targeting these molecules for the treatment of pancreatic cancer.


Epstein-Barr virus-induced sickle hepatopathy.
Sickle hepatopathy comprises a spectrum of disorders that vary in severity. Intravascular sickling and sinusoidal occlusion are the principal drivers of sickle hepatopathy, but infection or autoimmunity can act as triggers. We describe two cases of acute sickle hepatopathy initiated by primary Epstein-Barr virus (EBV) infection, a previously unreported association. The first case entailed a 14-year-old girl with hemoglobin SC (HbSC) disease who developed hepatic sequestration crisis that responded to a simple transfusion of erythrocytes. The second case was that of a 16-year-old boy with HbSC disease who experienced life-threatening intrahepatic cholestasis with multiorgan failure.


Adsorption of tetracycline antibiotic onto modified zeolite: Experimental investigation and modeling.
Artificial Neural Networks (ANNs) model and Adaptive Neuro-Fuzzy Inference System (ANFIS) were used to estimate and predict the removal efficiency of tetracycline (TC) using the adsorption process from aqueous solutions. The obtained results demonstrated that the optimum condition for removal efficiency of TC were 1.5 g L-1 modified zeolite (MZ), pH of 8.0, initial TC concentration of 10.0 mg L-1, and reaction time of 60 min. Among the different back-propagation algorithms, the Marquardt-Levenberg learning algorithm was selected for ANN Model. The log sigmoid transfer function (log sig) at the hidden layer with ten neurons in the first layer and a linear transfer function were used for prediction of the removal efficiency. Accordingly, a correlation coefficient, mean square error, and absolute error percentage of 0.9331, 0.0017, and 0.56% were obtained for the total dataset, respectively. The results revealed that the ANN has great performance in predicting the removal efficiency of TC.•ANNs used to estimate and predict tetracycline antibiotic removal using the adsorption process from aqueous solutions.•The model's predictive performance evaluated by MSE, MAPE, and R2.


[Contribution to the knowledge of intestinal protozoa infestation in the hospital population of Barcelona (author's transl)].
This report concerns the analysis of 650 fecal samples submitted for parasitic investigation to the Microbiology Service of the Hospital de la Santa Creu i Sant Pau of Barcelona. The samples were analyzed by the technique of fixation and direct observation of Sapero and Lawless (MIFD), and with the biphasic concentration method of Blagg et al (MIFC). Intestinal protozoa were detected in 183 samples (28.2%), with a total of 232 protozoosis uncovered. The specific analysis gave the following results: Giardia lamblia, 10.9%; Dientamoeba fragilis, 7.8%; Entamoeba coli, 4.6%; Endolimax nana, 4.5%; Entamoeba hartmanni, 4.0%; Iodamoeba buetschlii, 2.9%; Entamoeba histolytica, 0.7%; Chilomastix mesnili, 0.2%; Trichomonas hominis, 0.2%. The results obtained by the two methods have been compared, observing the highest number of positive results with MIFD technique. Especial mention is made of the higher incidence of Dientamoeba fragilis in samples positive for Oxyuris further studies being needed to establish the basis for the relationship between both species of parasites.


The Canadian National Retirement Risk Index: employing statistics Canada's LifePaths to measure the financial security of future Canadian seniors.
This article measures a Canadian National Retirement Risk Index (NRRI). Originally developed by the Center for Retirement Research at Boston College, the NRRI is a forward-looking measure that evaluates the proportion of working-aged individuals who are at risk of not maintaining their standard of living in retirement. The Canadian retirement income system has been very effective in reducing elderly poverty, but our results suggest that it has been much less successful in maintaining the living standards of Canadians after retirement. Since the earlier years of the new millennium, we find that approximately one-third of retiring Canadians have been unable to maintain their working-age consumption after retirement—a trend that is projected to worsen significantly for future Canadian retirees. The release of the Canadian NRRI is timely given the widespread concern that the current Canadian retirement income system is inadequate. Many proposals have recently emerged to extend and/or enhance Canadian public pensions, and the NRRI is a tool to test their merit. The methodology underlying the Canadian NRRI is uniquely sophisticated and comprehensive on account of our employment of Statistics Canada's LifePaths, a state-of-the-art stochastic microsimulation model of the Canadian population. For instance, the Canadian NRRI is novel in that it models all of the relevant sources of consumption before and after retirement, while accounting for important features that are typically neglected in retirement adequacy studies such as family size, the variation of consumption over a person's lifetime, and the heterogeneity among the life courses of individuals.


Aging is associated with quantitative and qualitative changes in circulating cell-free DNA: the Vitality 90+ study.
As a marker of cellular death, cell-free DNA (cf-DNA) has a utility in diagnosis and prognosis of various disorders. Since aging accompanies increased cellular senescence and death, we aimed to characterize potential age-related alterations in cf-DNA. The study population consisted of 12 nonagenarian women (participants in the Vitality 90+ Study) and 11 healthy control women (aged 22-37 years). Some of the nonagenarians (n=8) were also recruited for follow-up after one year. cf-DNA was extracted using two different methods. Total cf-DNA was quantified directly in plasma and the amplifiable cf-DNA was assessed using quantitative PCR. Quality of cf-DNA was analysed with a DNA Chip assay. For all the quantification methods, the concentration of cf-DNA was significantly higher (p<0.05) in nonagenarians as compared to controls. The quality of the cf-DNA also displayed a marked difference between nonagenarians and controls; a fragmented pattern or appearance of low molecular weight cf-DNA was observed in the majority of the nonagenarians, whereas in controls, cf-DNA was intact and had a quasi-genomic, high molecular weight appearance. In nonagenarians, the quality of cf-DNA appeared similar in the original and follow-up samples. We propose that some, as yet uncharacterized, aspects of aging are reflected in the appearance of cf-DNA.


Deletion of Coagulation Factor IX Compromises Bone Mass and Strength: Murine Model of Hemophilia B (Christmas Disease).
Osteopenia and osteoporosis have increasingly become a recognized morbidity in those persons with hemophilia (PwH) receiving inadequate prophylactic clotting factor replacement. Animal models can control or eliminate genetic and environmental factors and allow for invasive testing not clinically permissible. Here, we describe the skeletal phenotype of juvenile and adult male mice with a genetically engineered deficiency in coagulation factor IX (FIX KO). Although the somatic growth of FIX KO mice matched that of their wild-type (WT) littermates at 10 and 20 weeks of age, the FIX KO mice displayed reduced bone mineral density (BMD), reduced cortical and cancellous bone mass, and diminished whole bone fracture resistance. These findings coupled with parallel observations in a murine model of hemophilia A (FVIII deficiency) point to an effector downstream of the coagulation cascade that is necessary for normal skeletal development. Further study of potential mechanisms underlying the bone disease observed in rare clotting factor deficiency syndromes may lead to new diagnostic and therapeutic insights for metabolic bone diseases in general.