Association Between 6-Week Postdischarge Risk Classification and 12-Month Outcomes After Orthopedic Trauma.
Numerous studies have demonstrated that long-term outcomes after orthopedic trauma are associated with psychosocial and behavioral health factors evident early in the patient's recovery. Little is known about how to identify clinically actionable subgroups within this population. To examine whether risk and protective factors measured at 6 weeks after injury could classify individuals into risk clusters and evaluate whether these clusters explain variations in 12-month outcomes. A prospective observational study was conducted between July 16, 2013, and January 15, 2016, among 352 patients with severe orthopedic injuries at 6 US level I trauma centers. Statistical analysis was conducted from October 9, 2017, to July 13, 2018. At 6 weeks after discharge, patients completed standardized measures for 5 risk factors (pain intensity, depression, posttraumatic stress disorder, alcohol abuse, and tobacco use) and 4 protective factors (resilience, social support, self-efficacy for return to usual activity, and self-efficacy for managing the financial demands of recovery). Latent class analysis was used to classify participants into clusters, which were evaluated against measures of function, depression, posttraumatic stress disorder, and self-rated health collected at 12 months. Among the 352 patients (121 women and 231 men; mean [SD] age, 37.6 [12.5] years), latent class analysis identified 6 distinct patient clusters as the optimal solution. For clinical use, these clusters can be collapsed into 4 groups, sorted from low risk and high protection (best) to high risk and low protection (worst). All outcomes worsened across the 4 clinical groupings. Bayesian analysis shows that the mean Short Musculoskeletal Function Assessment dysfunction scores at 12 months differed by 7.8 points (95% CI, 3.0-12.6) between the best and second groups, by 10.3 points (95% CI, 1.6-20.2) between the second and third groups, and by 18.4 points (95% CI, 7.7-28.0) between the third and worst groups. This study demonstrates that during early recovery, patients with orthopedic trauma can be classified into risk and protective clusters that account for a substantial amount of the variance in 12-month functional and health outcomes. Early screening and classification may allow a personalized approach to postsurgical care that conserves resources and targets appropriate levels of care to more patients.


Pharmacological strategies in depression.
Several pharmacological manoeuvres may be considered when the first attempt at antidepressant treatment fails. The importance of adequate dosage and duration in any treatment trial are emphasized, and the evidence in support of combined antidepressants, lithium-antidepressant combinations, the addition of triiodothyronine or tryptophan, the use of anticonvulsants, and the use of vanadium inactivators, is evaluated.


Use of in vivo expression technology to identify genes important in growth and survival of Pseudomonas fluorescens Pf0-1 in soil: discovery of expressed sequences with novel genetic organization.
Studies were undertaken to determine the genetic needs for the survival of Pseudomonas fluorescens Pf0-1, a gram-negative soil bacterium potentially important for biocontrol and bioremediation, in soil. In vivo expression technology (IVET) identified 22 genes with elevated expression in soil relative to laboratory media. Soil-induced sequences included genes with probable functions of nutrient acquisition and use, and of gene regulation. Ten sequences, lacking similarity to known genes, overlapped divergent known genes, revealing a novel genetic organization at those soil-induced loci. Mutations in three soil-induced genes led to impaired early growth in soil but had no impact on growth in laboratory media. Thus, IVET studies have identified sequences important for soil growth and have revealed a gene organization that was undetected by traditional laboratory approaches.


Neutralization of Tumor Acidity Improves Antitumor Responses to Immunotherapy.
Cancer immunotherapies, such as immune checkpoint blockade or adoptive T-cell transfer, can lead to durable responses in the clinic, but response rates remain low due to undefined suppression mechanisms. Solid tumors are characterized by a highly acidic microenvironment that might blunt the effectiveness of antitumor immunity. In this study, we directly investigated the effects of tumor acidity on the efficacy of immunotherapy. An acidic pH environment blocked T-cell activation and limited glycolysis in vitro. IFNγ release blocked by acidic pH did not occur at the level of steady-state mRNA, implying that the effect of acidity was posttranslational. Acidification did not affect cytoplasmic pH, suggesting that signals transduced by external acidity were likely mediated by specific acid-sensing receptors, four of which are expressed by T cells. Notably, neutralizing tumor acidity with bicarbonate monotherapy impaired the growth of some cancer types in mice where it was associated with increased T-cell infiltration. Furthermore, combining bicarbonate therapy with anti-CTLA-4, anti-PD1, or adoptive T-cell transfer improved antitumor responses in multiple models, including cures in some subjects. Overall, our findings show how raising intratumoral pH through oral buffers therapy can improve responses to immunotherapy, with the potential for immediate clinical translation.


[Epilepsy and the brain's morphological-functional organization].
The article summarized the results of long-term studies of the author with using experimental methods--creation of penicillin and aluminium epileptogenic foci, orbito-frontal cortex stimulation and elimination, and clinical investigation of epileptic status, prefrontal epilepsy, absence EEG activity, symptomatic rolandic epilepsy, childhood epilepsy. Based on the results obtained, the author formulates the conceptions on (1) the key role of prefrontal cortex in the antiepileptic defense system and motor activity control, (2) the role of slow activity in antiepileptogenesis, (3) the age-dependent rhythmic possibilities of children's brain, which determined age-dependent epilepsy types, (4) the complexity of between-hemispheric interactions in this disease.


Nutritive value and fermentation characteristics of alfalfa-mixed grass forage wrapped with minimal stretch film layers and stored for different lengths of time.
A key aspect of managing baled silages is to quickly achieve and then rigorously maintain anaerobic conditions within the silage mass. The concept of inserting an O2-limiting barrier (OB) into plastic commercial silage wraps has been evaluated previously, yielding mixed or inconclusive results. Our objective for this study was to maximize the challenge to a commercial polyethylene bale wrap, or the identical wrap containing an OB, by using minimal plastic (4 layers), and then extending storage periods as long as 357 d. Forty-eight 1.2 × 1.2-m large-round bales of alfalfa (Medicago sativa L.) and mixed grass forage (66.3 ± 8.66% alfalfa; DM basis) were made at 2 moisture concentrations [47.5 (ideal) or 36.1% (dry)], wrapped with 4 layers of plastic containing an OB or no OB, and then stored for 99, 243, or 357 d. After storage, yeast counts within the 0.15-m deep surface layer were not affected by treatment (mean = 5.85 log10 cfu/g); mold counts could not be analyzed statistically because 26 bales were nondetectable at a 3.00 log10 cfu/g detection limit, but means among detectable counts were numerically similar for OB (4.74 log10 cfu/g) and no OB (4.77 log10 cfu/g). Fermentation characteristics were most affected by initial bale moisture, resulting in a more acidic final pH for ideal compared with dry bales (5.52 vs. 6.00). This was facilitated by greater concentrations of total fermentation acids (3.80 vs. 1.45% of dry matter), lactic acid (2.24 vs. 0.71% of dry matter), and acetic acid (1.07 vs. 0.64% of dry matter) within ideal compared with dry silages. Plastic wrap type had no effect on final concentrations of any fermentation product. During fermentation and storage, we noted greater change in concentrations of fiber components and whole-plant ash within the 0.15-m deep surface layer than in the bale core, and these changes always differed statistically from 0 (no change) based on pre-ensiled baseline concentrations. Overall, concentrations of water-soluble carbohydrates were reduced (mean = 2.3 percentage units) during fermentation and storage, which resulted (indirectly) in increased concentrations of fiber components and crude protein, as well as an overall energy cost of 2.2 percentage units of total digestible nutrient. It remains unclear under what conditions an OB plastic wrap will consistently benefit the fermentation and preservation of baled silages.


Studies on the mechanism of T cell inhibition by the Pseudomonas aeruginosa phenazine pigment pyocyanine.
Pseudomonas aeruginosa and its products have been shown to inhibit mitogen-induced human lymphocyte blastogenesis as measured by [3H]TdR uptake. The phenazine pigment pyocyanine has been identified as one of the inhibitors present in cellfree culture supernatants. To determine the mechanism of the inhibitory action of pyocyanine, we studied its effect on the early stages of T cell activation. Pyocyanine inhibited lymphocyte stimulation induced by specific antigens, the lectin concanavalin A and the calcium ionophore, ionomycin, suggesting that its inhibitory effect is not dependent on interference with the T cell antigen receptor complex itself. Using quin-2, we showed that pyocyanine did not interfere with the mitogen-induced increase in cytosolic-free Ca2+. We also showed that pyocyanine did not interfere with the function of calmodulin stimulated Ca2+-Mg2+ ATPase activity, indicating that the mechanism of action of pyocyanine differs from that of the structurally related phenothiazine compounds. Analysis of IL 2 production and IL 2 receptor expression clearly showed that pyocyanine inhibits the production of this essential lymphokine as well as the expression of IL 2 receptors on the T cell membrane. This inhibition is dose dependent and not due to cellular toxicity. There was parallel inhibition of growth in cell volume as well as [3H]TdR uptake. Thus, our results demonstrate that pyocyanine inhibits T cell proliferation by decreasing the production of the critical lymphokine IL 2 and by decreasing the expression of the IL 2 receptor. Local suppression of lymphocyte stimulation by phenazine pigments such as pyocyanine may interfere with cellular immune responses that may be necessary for eradication of chronic infection with P. aeruginosa.


Laparoscopic repair of traumatic abdominal wall hernia from handlebar injury.
A 14-year-old boy was seen at an outside hospital after falling over the handlebar of his bicycle and was discharged home. He was subsequently seen in our emergency department with complaints of persistent abdominal pain. A computed tomography scan of the abdomen revealed disruption of the muscles of the upper right abdominal wall containing the hepatic flexure of the colon, with a small amount of intraperitoneal free fluid noted. The patient underwent laparoscopic exploration using 3 ports (2-5 mm and 1-12 mm) and 2 separate stab incisions. The traumatic abdominal wall hernia was repaired with interrupted sutures placed with an ENDO CLOSE (Covidien, Mansfield, MA) device, and a mesenteric defect in the colon was approximated with intracorporeal sutures. The trocar sites were sutured closed. The patient recovered well and was discharged home. Follow-up examination revealed no abdominal wall defect and resolution of his symptoms. Laparoscopic repair of a traumatic abdominal wall defect and exploratory laparoscopy after trauma is feasible and safe in the pediatric patient. It should be considered as an alternative approach with potentially less morbidity than an exploratory laparotomy for handlebar injuries in a stable patient.


Nitric oxide (NO) and interleukin-1beta (IL-1beta) in follicular fluid and their correlation with fertilization and embryo cleavage.
Using an IVF model, the goal of the study was to investigate the relationship between follicular fluid (ff) NO and IL-1beta levels, as well as their correlation with fertilization of mature oocytes and embryo cleavage rates. Follicular fluid was collected from 17 patients at the time of transvaginal oocyte retrieval following controlled ovarian stimulation. Oocytes harvested from these follicles were followed through fertilization and embryo cleavage. The NO metabolites nitrate/nitrite (NO3/NO2) were measured using the Griess reaction as an indirect assessment of NO activity. IL-1beta was measured using a high sensitivity ELISA system (Amersham, UK). The Student's t-test was utilized for unpaired data with the means considered significantly different when P < or = 0.05. Follicular fluid NO3/NO2 levels were significantly lower in follicles containing mature oocytes that fertilized (n = 30; 9.7 +/- 1.0 microM), versus those that did not fertilize (n = 23; 15.4 +/- 2.4 microM; P < 0.05). Follicles that contained oocytes that fertilized and went on to divide beyond the 6 cell stage had significantly lower ff levels of NO3/NO2 (n = 18; 7.5 +/- 0.9 microM), as compared to ff that contained oocytes that did not fertilize or failed to develop beyond the 5 cell stage (n = 35; 14.6 +/- 1.7 microM; P < 0.01). No correlation was found between ff NO3/NO2 levels (n = 28; 13.8 +/- 2.0 microM) and ff IL-1beta levels (n = 28; 0.5 +/- 0.08 pg/mL). An analysis of ff IL-1beta levels in relation to fertilization and embryo cleavage rates revealed no correlation. Lower ff NO3/NO2 levels at the time of oocyte retrieval are associated with adequate fertilization and embryo cleavage rates. In our IVF model, no correlation was found between ff IL-1beta levels and ff NO3/NO2, fertilization, or embryo cleavage rates.


Pharmacophore-based virtual screening: a review of recent applications.
In research relating to the development of new drugs, hit identification and validations are critical for successful optimization of candidates. To achieve rapid identification of new lead compounds, high-throughput screening assays have been employed in many pharmaceutical companies and laboratories. However, their success depends on the assay system relevant to in vivo conditions and they are physically limited by the repertoire of compounds. As an alternative or complementary approach to high-throughput screening assays, virtual screening is an efficient method to identify drug candidates in silico from large chemical compound databases. Its usefulness has been verified by current applications that successfully retrieved hit and lead identifications against various disease targets. However, for better application, the scoring functions for distinguishing possible active and inactive compounds must beimproved. In this review, we provide an overview of pharmacophore-based virtual screening methods with a special focus on their successful application towards finding hits against various diseasetargets. Readers will rapidly gain insight into the recent successful applications of pharmacophore-based virtual screening. They will acknowledge that this technique is a powerful and cost-effective alternative to high-throughput assays. Although there are many hurdles yet to be resolved, virtual screening techniques will emerge as essential infrastructure and as a prerequisite for developing new lead compounds with therapeuticapplications.


Apple-peel atresia presenting as foetal intestinal obstruction.
Apple-peel atresia or Type 3 jejuno-ileal atresia (JIA) is an uncommon cause of foetal intestinal obstruction. Bowel obstruction in the foetus is diagnosed on the prenatal ultrasonography only in 50% cases. We report a case in which foetal intestinal obstruction was diagnosed on prenatal ultrasonography. The child showed signs of intestinal obstruction on day one after birth, for which an exploratory laparotomy was performed. Type 3 JIA was found for which resection of atretic segments with jejuno-ascending colon anastomosis was preformed.


Lapatinib versus hormone therapy in patients with advanced renal cell carcinoma: a randomized phase III clinical trial.
Lapatinib is an orally reversible inhibitor of epidermal growth factor receptor (EGFR)/human epidermal growth factor receptor 2 (HER-2) tyrosine kinases with demonstrated activity in patients with HER-2-positive breast cancer. In the current phase III open-label trial, lapatinib was compared with hormone therapy (HT) in patients with advanced renal cell carcinoma (RCC) that express EGFR and/or HER-2. Patients with advanced RCC who had experienced disease progression through first-line cytokine therapy--stratified by Karnofsky performance status and number of metastatic sites--were randomly assigned to lapatinib 1,250 mg daily or HT. The primary end point was time to progression (TTP); secondary end points included overall survival (OS), safety, and biomarker analyses. Four hundred sixteen patients were enrolled onto the study. Median TTP was 15.3 weeks for lapatinib versus 15.4 weeks for HT (hazard ratio [HR] = 0.94; P = .60), and median OS was 46.9 weeks for lapatinib versus 43.1 weeks for HT (HR = 0.88; P = .29). In a biomarker analysis of patients with EGFR-overexpressed tumors (3+ by immunohistochemistry [IHC]; n = 241) median TTP was 15.1 weeks for lapatinib versus 10.9 weeks for HT (HR = 0.76; P = .06), and median OS was 46.0 weeks for lapatinib versus 37.9 weeks for HT (HR = 0.69; P = .02). These results were confirmed by Cox regression analysis. No unexpected toxicities were observed; the most commonly reported drug-related adverse events (all grades) for lapatinib were rash (44%) and diarrhea (40%). Lapatinib was well tolerated with equivalent overall efficacy to HT in advanced RCC patients who had experienced disease progression while receiving cytokines, and the study supports that lapatinib prolonged OS relative to HT in patients with 3+ EGFR status determined by IHC.


Visceral adipose tissue but not subcutaneous adipose tissue is associated with urine and serum metabolites.
Obesity is a complex multifactorial phenotype that influences several metabolic pathways. Yet, few studies have examined the relations of different body fat compartments to urinary and serum metabolites. Anthropometric phenotypes (visceral adipose tissue (VAT), subcutaneous adipose tissue (SAT), the ratio between VAT and SAT (VSR), body mass index (BMI), waist circumference (WC)) and urinary and serum metabolite concentrations measured by nuclear magnetic resonance spectroscopy were measured in a population-based sample of 228 healthy adults. Multivariable linear and logistic regression models, corrected for multiple testing using the false discovery rate, were used to associate anthropometric phenotypes with metabolites. We adjusted for potential confounding variables: age, sex, smoking, physical activity, menopausal status, estimated glomerular filtration rate (eGFR), urinary glucose, and fasting status. In a fully adjusted logistic regression model dichotomized for the absence or presence of quantifiable metabolite amounts, VAT, BMI and WC were inversely related to urinary choline (ß = -0.18, p = 2.73*10-3), glycolic acid (ß = -0.20, 0.02), and guanidinoacetic acid (ß = -0.12, p = 0.04), and positively related to ethanolamine (ß = 0.18, p = 0.02) and dimethylamine (ß = 0.32, p = 0.02). BMI and WC were additionally inversely related to urinary glutamine and lactic acid. Moreover, WC was inversely associated with the detection of serine. VAT, but none of the other anthropometric parameters, was related to serum essential amino acids, such as valine, isoleucine, and phenylalanine among men. Compared to other adiposity measures, VAT demonstrated the strongest and most significant relations to urinary and serum metabolites. The distinct relations of VAT, SAT, VSR, BMI, and WC to metabolites emphasize the importance of accurately differentiating between body fat compartments when evaluating the potential role of metabolic regulation in the development of obesity-related diseases, such as insulin resistance, type 2 diabetes, and cardiovascular disease.


Axillary lymph nodes: mammographic, pathologic, and clinical correlation.
The purpose of this study was to determine the cause and frequency of axillary abnormalities seen mammographically and to evaluate the imaging characteristics of lymphadenopathy that are associated with malignancy. Ninety-six axillary abnormalities seen mammographically in 94 patients were retrospectively reviewed and correlated with the clinical diagnoses and pathologic results found in the medical records. For each abnormality, the length, margins, and presence of microcalcifications were noted. Logistic regression was used to determine an association between these findings and status (benign or malignant). Seventy-six of 94 patients had lymphadenopathy. Eighteen of 94 patients had an abnormality other than lymphadenopathy. Because two of these 94 patients had more than one abnormality, a total of 96 abnormalities occurred, 20 of which were due to an abnormality other than lymphadenopathy. Regarding the 76 cases of lymphadenopathy, the most frequent diagnosis was nonspecific benign lymphadenopathy in 29% (n = 22) of cases, followed by metastatic breast cancer in 26% (n = 20) and chronic lymphocytic leukemia or well-differentiated lymphocytic lymphoma in 17% (n = 13). Other causes (n = 21) included collagen vascular disease, lymphomas other than well-differentiated lymphocytic lymphoma, metastatic disease from nonbreast primary site, metastatic disease from unknown primary site, sarcoidosis. HIV-related lymphadenopathy, and reactive lymphadenopathy associated with a breast abscess. An association between length of nonfatty lymph nodes and malignant status was statistically significant at the .001 level. When a length greater than 33 mm was used as a predictor of malignancy, the specificity and sensitivity were 97% and 31%, respectively. We found an association between malignancy and nonfatty lymph nodes with ill-defined or spiculated margins (p = .053). Regarding the 20 abnormalities other than lymphadenopathy, epidermal cysts (n = 7) were most prevalent. The most common axillary abnormality revealed on mammography was abnormal lymph nodes. Homogeneously dense (nonfatty) axillary lymph nodes were strongly associated with malignancy when the lymph nodes were longer than 33 mm, had ill-defined or spiculated margins, or contained intranodal microcalcifications. However, our study confirmed that in most cases benign and malignant lymph nodes cannot be distinguished from each other mammographically.


Integrative Analysis of Transcriptomic and Proteomic Profiling in Inflammatory Bowel Disease Colon Biopsies.
Crohn's disease (CD) and ulcerative colitis (UC) are intestinal chronic inflammatory conditions characterized by altered epithelial barrier function and tissue damage. Despite significant efforts to understanding the biological mechanisms responsible for gut inflammation, the pathophysiology of CD and UC remains poorly understood. To help elucidate the potential mechanisms responsible for gut inflammation in CD and UC, transcriptomic and proteomic profiling of human colon biopsy specimens was performed. Dysregulated genes and proteins in disease tissues compared with normal tissues were characterized from the expression profiles and further subjected to pathway analysis to identify altered biological processes and signaling pathways. Sample analysis showed 4250 genes with matched protein expression and a wide range of correlation of RNA-protein abundance across samples. Pathway analysis of dysregulated genes and proteins in CD and UC showed alterations in immune and inflammatory responses, complement cascade, and the suppression of metabolic processes and PPAR signaling. In CD, increased T-helper cell differentiation and elevated toll-like receptor and JAK/STAT signaling were observed. Interestingly, increased MAPK signaling was only observed in UC. Weighted gene co-expression network analysis suggested a possible role of epigenetic regulation in UC. Of note, a large discrepancy between regulation of RNA and protein levels in inflamed colon samples was detected for previously identified biomarkers including MMP14 and LAMP1. With the analysis of dysregulated genes and pathways, the present study unravels key mechanisms contributing to CD and UC pathogenesis and emphasizes that integrative analysis of multi-omics data sets can provide more insight into understanding complex disease mechanisms.


Furosemide-loaded Alginate Microspheres Prepared by Ionic Cross-linking Technique: Morphology and Release Characteristics.
Furosemide-loaded alginate microspheres were prepared by the ionic cross-linking technique using CaCl(2), Al(2)(SO(4))(3) and BaCl(2). The process induced the formation of microspheres with the incorporation efficiency of 65% to 93%. The effect of sodium alginate concentration, cross-linking agents and drying conditions was evaluated with respect to entrapment efficiency, particle size, surface characteristics and in vitro release behaviors. Infrared spectroscopic study confirmed the absence of any drug-polymer interaction. Differential scanning calorimetric analysis revealed that the drug was molecularly dispersed in the alginate microspheres matrices showing rough surface, which was confirmed by scanning electron microscopy study. The mean particle size and entrapment efficiency were found to be varied by changing various formulation parameters. The in vitro release profile could be altered significantly by changing various formulation parameters to give a sustained release of drug from the microspheres. The kinetic modeling of the release data indicate that furosemide release from the alginate microspheres follow anomalous transport mechanism after an initial lag period when the drug release mechanism was found to be fickian diffusion controlled.


The influence of perioperative blood transfusion on survival after esophageal resection for carcinoma.
There is evidence that perioperative blood transfusion may lead to immunosuppression. Our aim was to determine whether blood transfusion influenced survival after esophagectomy for carcinoma. The study group comprised 234 consecutive patients (175 men and 59 women) with a mean age of 66 years who underwent esophagectomy for carcinoma by one surgeon between 1988 and 1998. The impact of 41 variables on survival was determined by means of univariate and multivariate analysis. Follow-up was complete (mean follow-up, 19.2 months; standard deviation, 16 months; range, 0 to 129 months). The operative mortality rate was 5.6% (13 deaths). Median operative blood loss was 700 mL (range, 150 to 7,000 mL). One hundred sixty-one patients (68.8%) received a blood transfusion postoperatively (mean transfusion, 2.6 units; range, 0 to 12 units). Overall actuarial 1-year, 3-year, and 5-year survival rates inclusive of operative mortality were 58.1%, 28.5%, and 16.1%, respectively. On univariate analysis, positive lymph nodes, pathological TNM stage, transfusion of more than 3 units of blood, incomplete resection, poor tumor cell differentiation, longer tumor, greater weight loss, male sex, and adenocarcinoma were significant (p < 0.05) negative factors for survival. On Cox proportional hazards regression analysis, after excluding operative mortality, lymph node involvement (p = 0.001), incomplete resection (p = 0.0001), poor tumor cell differentiation (p = 0.04), and transfusion of more than 3 units of blood (p = 0.04) were independent adverse predictors of late survival. In addition to reaffirming the importance of completeness of resection and nodal involvement, this study demonstrates that blood transfusion (more than 3 units) may have a significant adverse effect on late survival after esophageal resection for carcinoma. Every effort should be made to limit the amount of transfused blood to the absolutely essential requirements.


Acute otitis media caused by Streptococcus pyogenes in children.
Streptococcus pyogenes, or group A beta -hemolytic Streptococcus (GAS), is an important causative agent of bacterial pharyngotonsillititis and skin, soft-tissue, and invasive infections. Although it is also an important pathogen in acute otitis media (AOM), its exact role has not been determined. Patients aged 0-18 years with AOM, from whom a specimen of middle-ear fluid was obtained and cultured during 1999-2003, were enrolled. Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, and GAS were considered pathogens. Information collected included demographic characteristics, clinical history, and signs and symptoms. GAS otitis was observed in 350 (3.1%) of 11,311 episodes (of which 117 were also culture-positive for other pathogens). The other 10,961 episodes involved H. influenzae only (n = 2507), S. pneumoniae only (n = 2131), dual infection with H. influenzae and S. pneumoniae (n = 1290), M. catarrhalis only (n = 129), and other combinations of pathogens (n = 271). Increased age and Jewish ethnicity were independent, significant, positive risk factors for GAS AOM, and fall season was a negative risk factor. Episodes of GAS infection were less frequently bilateral, febrile, and accompanied by other systemic findings than were other episodes of other types of infection. Most patients with GAS AOM presented with acute drainage from the ears. A lower proportion of cases of AOM were due to GAS in children with recurrent AOM and in patients recently treated with antibiotics, compared with patients with AOM who did not have these factors. The risk for mastoiditis was highest among patients with GAS AOM, compared with patients infected with other pathogens: 11.6 episodes per 1000 episodes of GAS AOM, compared with 2.2, 0.3, and 0 episodes of mastoiditis per 1000 episodes of AOM due to S. pneumoniae, H. influenzae, and M. catarrhalis, respectively. Compared with AOM caused by pathogens other than GAS, GAS AOM is characterized by older age and higher local aggressiveness manifested by lower rates of fever and respiratory symptoms and higher rates of tympanic perforation and mastoiditis.


Isolation and characterization of low-sulphur-tolerant mutants of Arabidopsis.
Sulphur is an essential element for plant growth and development as well as for defence against biotic and abiotic stresses. Increasing sulphate utilization efficiency (SUE) is an important issue for crop improvement. Little is known about the genetic determinants of sulphate utilization efficiency. No gain-of-function mutants with improved SUE have been reported to date. Here the isolation and characterization of two low-sulphur-tolerant mutants, sue3 and sue4 are reported using a high-throughput genetic screen where a 'sulphur-free' solid medium was devised to give the selection pressure necessary to suppress the growth of the wild-type seedlings. Both mutants showed improved tolerance to low sulphur conditions and well-developed root systems. The mutant phenotype of both sue3 and sue4 was specific to sulphate deficiency and the mutants displayed enhanced tolerance to heavy metal and oxidative stress. Genetic analysis revealed that sue3 was caused by a single recessive nuclear mutation while sue4 was caused by a single dominant nuclear mutation. The recessive locus in sue3 is the previously identified VirE2-interacting Protein 1. The dominant locus in sue4 is a function-unknown locus activated by the four enhancers on the T-DNA. The function of SUE3 and SUE4 in low sulphur tolerance was confirmed either by multiple mutant alleles or by recapitulation analysis. Taken together, our results demonstrate that this genetic screen is a reasonable approach to isolate Arabidopsis mutants with improved low sulphur tolerance and potentially with enhanced sulphate utilization efficiency. The two loci identified in sue3 and sue4 should assist in understanding the molecular mechanisms of low sulphur tolerance.


Validation of the Applicability of In-Cell Fast Photochemical Oxidation of Proteins across Multiple Eukaryotic Cell Lines.
Fast photochemical oxidation of proteins (FPOP), a hydroxyl radical-based protein footprinting method, coupled to mass spectrometry has been extensively used to study protein structure and protein-protein interactions in vitro. This method utilizes hydroxyl radicals to oxidatively modify solvent-accessible amino acids and has recently been demonstrated to modify proteins within live cells (IC-FPOP) and Caenorhabditis elegans. Here, we have expanded the application of IC-FPOP into a variety of commonly used cell lines to verify the applicability of the method across various cellular systems. IC-FPOP was able to successfully modify proteins in five different cell lines (Vero, HEK 293T, CHO, MCF-10A, and MCF-7). To increase the number of oxidatively modified proteins identified, we have also employed the use of offline high pH reversed-phase liquid chromatography (RPLC) followed by concatenation and online low-pH RPLC. The coupling of IC-FPOP to 2D-LC MS/MS resulted in a 1.7-fold increase in total identifications of oxidatively modified proteins, which expanded the dynamic range of the method. This work demonstrates the efficacy of using IC-FPOP to study protein-protein interactions in cells.


[Adrenal gland lymphangiomas. A case report].
Adrenal gland lymphangiomas are benign, non-functional, and uncommon tumors of lymphatic tissue and with a cyst component. The majority of these are asymptomatic and are discovered incidentally. We present the case of an adrenal gland lymphangioma in a 24-year-old female with a history of high blood pressure, headaches, and anxiety of nearly 2 years in whom a routine ultrasound exam revealed a cyst tumor of right adrenal gland confirmed by computer tomography (CT) scan. Surgery was performed and lymphangioma of adrenal gland was the pathologist's report. The patient's clinical symptoms disappeared.


Ectopic expression of an orchid (Oncidium Gower Ramsey) AGL6-like gene promotes flowering by activating flowering time genes in Arabidopsis thaliana.
An AP1/AGL9 group of MADS box gene, OMADS1, with extensive homology to the Arabidopsis AGAMOUS-like 6 gene (AGL6) was characterized from orchid (Oncidium Gower Ramsey). OMADS1 mRNA was detected in apical meristem and in the lip and carpel of flower. Yeast two-hybrid analysis indicated that OMADS1 is able to strongly interact with OMADS3, a TM6-like protein that was involved in flower formation and floral initiation in orchid. Transgenic Arabidopsis and tobacco ectopically expressed OMADS1 showed similar novel phenotypes by significantly reducing plant size, flowering extremely early, and losing inflorescence indeterminacy. In addition, homeotic conversion of sepals into carpel-like structures and petals into staminoid structures were also observed in flowers of 35S::OMADS1 Arabidopsis. This result indicated that OMADS1 was involved in floral formation and initiation in transgenic plants. Further analysis indicated that the expression of flowering time genes FT, SUPPRESSOR OF OVEREXPRESSION OF CO 1 (SOC1) and flower meristem identity genes LEAFY (LFY), APETALA1 (AP1) was significantly up-regulated in 35S::OMADS1 transgenic Arabidopsis plants. Furthermore, ectopic expression of OMADS1 rescued late-flowering phenotype in gi-1, co-3 but not for ft-1 and fwa-1 mutants. These results supported that ectopic expression of OMADS1 influenced flower transition and formation by acting as an activator for FT and SOC1 in Arabidopsis.


Trends, Predictors, and Outcomes of Temporary Mechanical Circulatory Support for Postcardiac Surgery Cardiogenic Shock.
Postcardiac surgery cardiogenic shock (PCCS) is seen in 2% to 6% of patients who undergo cardiac surgery. There are limited large-scale data on the use of mechanical circulatory support (MCS) in these patients. This study sought to evaluate the in-hospital mortality, trends, and resource utilization for PCCS admissions with and without MCS. A retrospective cohort of PCCS between 2005 and 2014 with and without the use of temporary MCS was identified from the National Inpatient Sample. Admissions for permanent MCS and heart transplant were excluded. Propensity-matching for baseline characteristics was performed. The primary outcome was in-hospital mortality and secondary outcomes included trends in use, hospital costs and lengths of stay. In the period between 2005 and 2014, there were 132,485 admissions with PCCS, with 51.3% requiring MCS. The intra-aortic balloon pump was the predominant device used with a steady increase in other devices. MCS use for more frequent in younger patients, males and those with higher co-morbidity. There was a decrease in MCS use across all demographic categories and hospital characteristics over time. Older age, female sex, previous cardiovascular morbidity and MCS use were independently predictive of higher in-hospital mortality. In 6,830 propensity-matched pairs, PCCS admissions that required MCS use, had higher in-hospital mortality (odds ratio 2.4; p<0.001), higher hospital costs ($98,759 ± 907 vs $81,099 ± 698; p<0.001) but not a longer length of stay compared with those without MCS use. In conclusion, in patients with PCCS, this study noted a steady decrease in MCS use. Use of MCS identified PCCS patients at higher risk for in-hospital mortality and greater resource utilization.


Equitable reach: Patient and professional recommendations for interventions to prevent perinatal depression and anxiety.
Perinatal depression and anxiety are the most common complications in the perinatal period and disproportionately affect those experiencing economic marginalization. Fewer than 15% of individuals at risk for perinatal depression are referred for preventative counseling. The goal of this study was to elicit patient and perinatal care professionals' perspectives on how to increase the reach of interventions to prevent perinatal depression and anxiety among economically marginalized individuals. We conducted qualitative interviews with perinatal individuals with lived experience of perinatal depression and/or anxiety who were experiencing economic marginalization (n = 12) and perinatal care professionals and paraprofessionals (e.g., obstetrician/gynecologists, midwives, doulas; n = 12) serving this population. Three study team members engaged a "a coding consensus, co-occurrence, and comparison," approach to code interviews. Perinatal individuals and professionals identified prevention intervention delivery approaches and content to facilitate equitable reach for individuals who are economically marginalized. Factors influential included availability of mental health counselors, facilitation of prevention interventions by a trusted professional, digital health options, and options for mental health intervention delivery approaches. Content that was perceived as increasing equitable intervention reach included emphasizing stigma reduction, using cultural humility and inclusive materials, and content personalization. Leveraging varied options for mental health intervention delivery approaches and content could reach perinatal individuals experiencing economic marginalization and address resource considerations associated with preventative interventions.


Quantitative analysis of wild-type and precore mutant hepatitis B virus in carriers.
In the present study, we have analyzed the amount of precore wild-type hepatitis B virus (HBV) (wild-type) and precore mutant HBV (nt 1896: G-->A) (precore mutant) of HBV carriers; 31 asymptomatic healthy carriers (ASC) and 28 patients with chronic hepatitis (CH). Wild-type and precore mutant were quantified using sensitive and specific quantification methods: competitive wild-type-sequence-specific assay and competitive mutation-site-specific assay with different sets of specific primers and internal controls. Median serum levels of wild-type and precore mutant were 9.60 and 8.60 log copies/ml (median percentages of precore mutant in total HBV-DNA: 11.7%) in HBeAg(+) ASC, 8.48 and 8.00 (33.3%) in HBeAg(+) CH, and 6.30 and 6.85 (84.7%) in anti-HBe(+) CH, respectively, showing higher levels of the relative amount of precore mutant to wild-type along with HBeAg/anti-HBe status. Only precore mutant, but not wild-type was detected in anti-HBe(+) ASC. Although median percentages of precore mutant at the anti-HBe(+) ASC and CH stages were much higher than those at the HBeAg(+) ASC and CH stages, a substantial amount of precore mutant was found even at the HBeAg(+) stages. Existence of a substantial amount of precore mutant even in HBeAg(+) ASC suggests that the occurrence of precore mutant is not always closely associated with seroconversion from HBeAg to anti-HBe.


Measurements of cardiac output in seriously ill patients using a CO2 rebreathing method.
A CO2 rebreathing method for the measurement of cardiac output was evaluated by comparison with the direct Fick O2 method in 26 studies performed in 18 patients who were critically ill. The method requires measurement of CO2 output by collection of expired gas, of arterial PCO2, and of mixed venous PCO2 by rebreathing. Twenty-five comparisons were within +/- 20% of the direct Fick measurements, at cardiac outputs varying between 1.4 and 6.4 L/min. Knowledge of the cardiac output increased the quality of the interpretation of arterial blood PO2 measurements in the assessment of pulmonary gas exchange disturbances.


Di-2-ethylhexylphthalate promotes thyroid cell proliferation and DNA damage through activating thyrotropin-receptor-mediated pathways in vitro and in vivo.
Phthalates are being suggested to be associated with altered thyroid function and proliferative changes, but detailed mechanisms remain unclear. Here, we examined the effects of di-(2-ethylhexyl) phthalate (DEHP) on DNA damage and proliferation in thyroid using thyroid carcinoma cell line, 8505C, in vitro and the rats orally treated with DEHP at 0, 0.3, 3, 30 and 150 mg/kg for 90 days from post-natal day 9 in vivo. Exposure to DHEP (1-50 μM) induced cellular proliferation, as evidenced by increased cell viability and DNA synthesis. Activation of γH2AX, a sensitive biomarker for DNA damage was observed following the exposure to DHEP (from 5 to 50 μM) with increased comet tail moment (5-100 μM) in comet assay, reflecting that DNA damage also occurred. When upstream signaling was examined, both thyrotropin receptor (TSHR)-ERK1/2 axis and TSHR-AKT axis were activated with upregulation of Pax8, a master transcriptional factor for thyroid differentiation and proliferation. Thyroid tissue from juvenile rats orally exposed to DEHP also confirmed DNA damage responses and the activation of TSHR signaling, which was evident from 0.3 to 3 mg/kg respectively. Notably, deletion of TSHR through siRNA attenuated these DEHP-induced events in vitro. Collectively these results suggest that DEHP induces DNA damage and cellular proliferation in thyroid, which appears to be from TSHR activation, providing an important insight into endocrine disrupting activities of phthalates on thyroid.


Simultaneous Real-Time Analysis of Bulk and Bottom Cure of Ultraviolet-Curable Inks Using Fourier Transform Infrared Spectroscopy.
The curing characteristics of an ultraviolet (UV) ink layer are of utmost importance for the development of UV inks. Measuring either bulk or bottom cure in itself is not new and has been the subject of many articles. In this article, two methods are described based on Fourier transform infrared (FT-IR) spectrometry to measure in real time and simultaneously the bulk and bottom cure of a thin UV ink layer. The procedure consists of applying a thin (10-12 µm) layer of UV-curing ink on an attenuated total reflection (ATR) crystal. The bottom cure is measured with ATR. The bulk cure is measured simultaneously with a reflection analysis (method 1) or a transmission analysis (method 2). With both methods, the bulk and bottom cure can be determined. To overcome problems with the interference in the ATR reflection setup, it is recommended to use the ATR transmission setup.


Shortening in femoral shaft fractures in children treated with spica cast.
Early spica cast treatment is one method used for children's femoral shaft fractures; it is increasingly advocated as treatment that allows early hospital discharge. The outcome of early spica cast treatment in 100 children, ages 2 to 10 years, with uncomplicated and isolated closed femoral shaft fractures treated at Johns Hopkins Hospital between October 1987 and March 1994 were analyzed. The objective was to identify those children who can be treated safely and dependably with early spica casting without excessive shortening of the fracture fragments. Eighty-one (81%) children had an acceptable outcome and 19 (19%) had an unacceptable outcome by the definition of more than 25 mm of fracture fragment overlap after clinical healing. A new clinical test, the telescope test, was statistically significant for correlation with spica cast outcome. Age, gender, fracture, location, mechanism of injury, fracture type, and resting radiograph of fracture fragment overlap were not statistically significant. The telescope test had a sensitivity of 80% and a specificity of 85% for predicting outcome. The relative risk for failure of spica cast treatment with a positive telescope test was 20.4 (95% confidence limits = 2.7-225.1). Children 2 to 10 years of age with uncomplicated femoral shaft fractures and a negative telescope test can be treated appropriately in most cases with early application of a spica cast.


Child care and common communicable illnesses: results from the National Institute of Child Health and Human Development Study of Early Child Care.
To examine the relationship between experiences in child care and communicable illnesses (gastrointestinal tract illness, upper respiratory tract infection, and ear infections or otitis media) throughout the first 3 years of life and to investigate whether increased frequency of these illnesses is related to language development, school readiness, and behavior problems. Health, child care, family, and child developmental data were obtained from more than 1200 participants in the National Institute of Child Health and Human Development Study of Early Child Care, a 10-site prospective study that began at the participants' birth. Longitudinal logistic regression analyses were performed using each type of communicable illness as the outcome variable, with family, child, and child care variables as predictors in the model, and followed by a series of regression analyses with developmental measures as the outcome variables. Rates of illness were higher in children in child care than for children reared exclusively at home during the first 2 years of life, but the differences were nonsignificant by age 3 years. Number of hours in child care per week during the first year and number of other children in the child care arrangement were related to the rates of illness. There was no evidence that increased rates of illness have a negative effect on school readiness or language competence. However, there was some evidence that increased illness was associated with behavior problems as reported by mothers, but not by child care providers. Children in child care experience more bouts of illness in the first 2 years of life, but differences are negligible by age 3 years. The increased rates of illness bear little relation to other aspects of children's development, except, perhaps, for a small increase in behavior problems.


Clinical signs in the Wernicke-Korsakoff complex: a retrospective analysis of 131 cases diagnosed at necropsy.
A recent necropsy study has shown that 80% of patients with the Wernicke-Korsakoff syndrome were not diagnosed as such during life. Review of the clinical signs of these cases revealed that only 16% had the classical clinical triad and 19% had no documented clinical signs. The incidence of clinical signs in this and other retrospective pathological studies is very different from that of prospective clinical studies. This discrepancy may relate to "missed" clinical signs but the magnitude of the difference suggests that at least some cases of the Wernicke-Korsakoff syndrome may be the end result of repeated subclinical episodes of vitamin B1 deficiency. In order to make the diagnosis, clinicians must maintain a high index of suspicion in the "at risk" group of patients, particularly alcoholics. Investigations of thiamine status may be helpful and if the diagnosis is suspected, parenteral thiamine should be given.


Long-term results of first-line sequential high-dose etoposide, ifosfamide, and cisplatin chemotherapy plus autologous stem cell support for patients with advanced metastatic germ cell cancer: an extended phase I/II study of the German Testicular Cancer Study Group.
Patients with disseminated germ cell cancer and poor prognosis (International Germ Cell Cancer Collaborative Group [IGCCCG] classification) achieve only a 45% to 50% long-term survival by standard chemotherapy. First-line high-dose chemotherapy might be able to improve the result. This analysis reports toxicity and long-term results of a large phase I/II study of sequential high-dose etoposide, ifosfamide, and cisplatin (VIP) in patients with advanced germ cell tumors. Between July 1993 and November 1999, 221 patients with either Indiana "advanced disease" (n = 39) or IGCCCG "poor prognosis" criteria (n = 182) received one cycle of VIP followed by three to four sequential cycles of high-dose VIP chemotherapy plus stem cell support, every 3 weeks, at six consecutive dose levels. Dose limiting toxicity occurred at level 8 (100 mg/m2 cisplatinum, 1750 mg/m2 etoposide, 12 g/m2 ifosfamide) with grade 4 mucositis (three of eight patients), grade 3 CNS toxicity (one of eight patients), grade 4 renal toxicity (one of eight patients), and prolonged granulocytopenia (one of eight patients). After 4-year median follow-up, progression-free survival and disease-specific survival rates in the poor prognosis subgroup were 69% and 79% at 2 years and 68% and 73% at 5 years, with 76% for gonadal/retroperitoneal versus 67% for mediastinal primaries. Severe toxicity included treatment related death (4%), treatment-related acute myeloid leukemia (1%), long-term impared renal function (3%), chronic renal failure (1%), and persistent grade 2-3 neuropathy (5%). Repetitive cycles of high-dose VIP with peripheral stem cell support can be successfully applied in a multicenter setting. Dose level 6 with cisplatin 100 mg/m2, etoposide 1500 mg/m2, and ifosfamide 10 g/m2 is recommended for further investigation in randomized trials. An ongoing randomized trial within the European Organization for Research and Treatment of Cancer evaluates this protocol against four cycles of standard cisplatin, etoposide, and bleomycin.


Heat shock protein 90 and ErbB2 in the cardiac response to doxorubicin injury.
A major drawback to doxorubicin as a cancer-treating drug is cardiac toxicity. To understand the mechanism of doxorubicin cardiac toxicity and the potent synergic effect seen when doxorubicin is combined with anti-ErbB2 (trastuzumab), we developed an in vivo rat model that exhibits progressive dose-dependent cardiac damage and loss of cardiac function after doxorubicin treatment. The hearts of these animals respond to doxorubicin damage by increasing levels of ErbB2 and the ErbB family ligand, neuregulin 1beta, and by activating the downstream Akt signaling pathway. These increases in ErbB2 protein levels are not due to increased ErbB2 mRNA, however, suggesting post-transcriptional mechanisms for regulating this protein in the heart. Accordingly, levels of heat shock protein 90 (HSP90), a known ErbB2 protein stabilizer and chaperone, are increased by doxorubicin treatment, and coimmunoprecipitation reveals binding of HSP90 to ErbB2. Isolated cardiomyocytes are more susceptible to doxorubicin after treatment with HSP90 inhibitor, 17-(allylamino)-17-demethoxygeldanamycin, suggesting that the HSP90 is protective during doxorubicin treatment. Thus, our results provide one plausible mechanism for the susceptibility of the heart to anti-ErbB2 therapy post-doxorubicin therapy in subclinical and clinical conditions. Additionally, these results suggest that further testing is needed for HSP90 inhibitors under various conditions in the heart.


Contingent Tunes of Neurochemical Ensembles in the Norm and Pathology: Can We See the Patterns?
Progress in the development of DSM/ICD taxonomies has revealed limitations of both label-based and dimensionality approaches. These approaches fail to address the contingent, nonlinear, context-dependent, and transient nature of those biomarkers linked to specific symptoms of psychopathology or to specific biobehavioural traits of healthy people (temperament). The present review aims to highlight the benefits of a functional constructivism approach in the analysis of neurochemical biomarkers underlying temperament and psychopathology. A review was performed. Eight systems are identified, and 7 neurochemical ensembles are described in detail. None of these systems is represented by a single neurotransmitter; all of them work in ensembles with each other. The functionality and relationships of these systems are presented here in association with their roles in action construction, with brief examples of psychopathology. The review introduces formal symbols for these systems to facilitate their more compact analysis in the future. This analysis demonstrates the possibility of constructivism-based unifying taxonomies of temperament (in the framework of the neurochemical model functional ensemble of temperament) and classifications of psychiatric disorders. Such taxonomies would present the biobehavioural individual differences as consistent behavioural patterns generated within a formally structured space of parameters related to the generation of behaviour.


[Intimate Partner Violence and Early Childhood DevelopmentViolência por parceiro íntimo e desenvolvimento na primeira infância].
To describe the prevalence of different types of intimate partner violence and estimate how this relates to child development in a low-income country in Latin America. The relationship between intimate partner violence and childcare practices, including the use of physical punishment, was also examined. An observational study was conducted using data collected as a baseline for an impact evaluation of the National Early Childhood Program in Nicaragua between 2013 and 2014. The sample included 8921 children between 0 and 5 years and 7436 mothers or caregivers in municipalities with an extreme poverty rate of over 0.2. The data revealed that 61% of mothers or caregivers reported controlling behavior by their partners, 50% suffered emotional violence, and 26% suffered physical violence at some point in their lives. Furthermore, the data demonstrated that children exposed to intimate partner violence displayed greater behavioral problems and delayed language and social-emotional development. These children are also more likely to be born prematurely and to have incomplete vaccination schedules. Finally, homes exposed to intimate partner violence are more likely to create more hostile and unsafe environments for children. The results reflect the magnitude of intimate partner violence and its detrimental effects on children. Public policies need to be devised and implemented not only to prevent this behavior and mitigate sequelae in exposed children but also to curb the intergenerational transmission of violence.


Lipoma arborescens; successfully treated by yttrium-90 radiosynovectomy.
Although radiosynovectomy (RS) applications have been carried out for many years, clinical indications of this non-invasive procedure is thought to be limited probably due to the lack of information of clinicians. Clinicians' preferential indication for RS is the treatment-resistant synovitis of individual joints, i.e. despite systemic pharmacotherapy and intra-articular steroid injections. We present here a case of "lipoma arborescens" treated by yttrium-90, which is a rare intra-articular lesion characterized by villous proliferation of the synovial membrane and hyperplasia of subsynovial fat. The results of clinical, biochemical and hematological examinations, magnetic resonance (MR) imaging, arthroscopy and histological analysis have shown that the etiology was lipoma arborescens in a female patient, aged 36 having swelling and sometimes associating pain at her right knee for 4 years. We have applied to our patient's right knee RS with 185 MBq yttrium-90 colloid together with 40 mg of methylprednisolone acetate, although in our literature survey we have not met any similar case being treated with such indication. Even a year after the application, the patient has absolutely benefited from the treatment clinically, and this was also confirmed by comparative MR images (pre- and post-treatment). Consequently, we consider that Y-90 treatment might be applicable in suitable cases with lipoma arborescens.


An update on immunotherapy options for urothelial cancer.
Introduction: The efficacy of immune checkpoint inhibitors has been shown in many malignancies. Urothelial cancers have a high mutational load and also express a high level of PD-L1. Therefore, the use of immune checkpoint inhibitors (ICIs) in various treatment lines is being intensively investigated in urothelial cancers.Area covered: In this review article, we aimed to summarize the development of immune checkpoint inhibition in urothelial cancers. We also provide updated and comprehensive data about the use of ICIs in the second-line and first-line treatment of patients who are ineligible for cisplatin. We also summarize ongoing trials and the results of published trials. A literature search was made using PubMed, Medline, and ASCO and ESMO Annual Meetings abstracts by using the following search terms: nivolumab, pembrolizumab, atezolizumab, avelumab, durvalumab, and urothelial cancer.Expert opinion: ICIs as a monotherapy are effective for the first-line treatment of the patients with advanced urothelial carcinoma who are ineligible for cisplatin, and also patients who are pre-treated with platinum-based chemotherapy. The results of trials investigating the efficacy of the combination of ICIs and chemotherapy in the first-line setting are awaited; it remains unclear as to whether this combination may become the standard first-line treatment in advanced urothelial cancer.


SEQUENCE SEGMENTATION USING JOINT RNN AND STRUCTURED PREDICTION MODELS.
We describe and analyze a simple and effective algorithm for sequence segmentation applied to speech processing tasks. We propose a neural architecture that is composed of two modules trained jointly: a recurrent neural network (RNN) module and a structured prediction model. The RNN outputs are considered as feature functions to the structured model. The overall model is trained with a structured loss function which can be designed to the given segmentation task. We demonstrate the effectiveness of our method by applying it to two simple tasks commonly used in phonetic studies: word segmentation and voice onset time segmentation. Results suggest the proposed model is superior to previous methods, obtaining state-of-the-art results on the tested datasets.


Alternatives for sample pre-treatment and HPLC determination of Ochratoxin A in red wine using fluorescence detection.
Ochratoxin A is a mycotoxin widely studied due to its nephrotoxic, immunotoxic, teratogenic and carcinogenic effects. The European Commission has fixed maximum limits for Ochratoxin A in wines and in other foods. In order to determine Ochratoxin A levels in red wine, the present paper contrasts and discusses the results of a systematic study of analytical parameters for sample pre-treatment using different immunoaffinity cartridges as well as C-18 cartridges with three solvent combinations. The direct injection of wine into two types of C-18 chromatographic columns (conventional packed column and monolithic column) is evaluated as screening method. In all cases, the analysis was carried out using HPLC with fluorescence detection. The results show statistical differences when 3 types of immunoaffinity columns were used, while higher recoveries were obtained for C-18 cartridges using acetonitrile as extraction solvent. Repeatability and accuracy of immunoaffinity and C-18 sample pre-treatment were statistically comparable (alpha=0.05). Their sensitivity was also comparable, although more favorable detection limits were obtained using the immunoaffinity treatment (0.01 microg L(-1)) in comparison with C-18 treatment (0.09 microg L(-1)). Considering the maximal allowed concentration of Ochratoxin A in wine (2.00 microg L(-1)), both methods are suitable for its determination in wine. Both methods were applied to determine this toxin in 154 wine samples, and the quantitative results demonstrated statistic comparability (alpha=0.05). These results were also confirmed from the qualitative point of view using a GC-MS method. To find an easy screening method, based on a recent publication, a monolithic HPLC column and 2 conventional packed columns were tested for Ochratoxin A determination in real wine samples by direct injection, without previous clean-up. The results show that this procedure is not useful at the concentration levels usually found in wine and although shorter time is required when using the monolithic columns even with the chromatographic analysis. Finally, based on the results, it was concluded that the combination of C-18 cartridges with conventional particle packed columns and HPLC-FLD is the most appropriate alternative for Ochratoxin A analysis in wine. Indeed, considering cost, sensitivity and selectivity, this method can be used in broad prospective programs.


Gate-tunable bolometer based on strongly coupled graphene mechanical resonators.
Bolometers based on graphene have demonstrated outstanding performance with high sensitivity and short response time. In situ adjustment of bolometers is very important in various applications, but it is still difficult to implement in many systems. Here we propose a gate-tunable bolometer based on two strongly coupled graphene nanomechanical resonators. Both resonators are exposed to the same light field, and we can measure the properties of one bolometer by directly tracking the resonance frequency shifts, and indirectly measure the other bolometer through mechanical coupling. We find that the sensitivity and the response bandwidth of both bolometers can be independently adjusted by tuning the corresponding gate voltages. Moreover, the properties of the indirectly measured bolometer show a dependence on the coupling between the two resonators, with other parameters being fixed. Our method has the potential to optimize the design of large-scale bolometer arrays, and open new horizons in infrared/terahertz astronomy and communication systems.


Time-resolved crystallography.
Time-resolved crystallography has been successfully applied on the time scale from seconds via milliseconds and nanoseconds to picoseconds on a variety of systems. This brief review largely deals with macromolecular systems, on which there has been substantial recent progress. The strategies for design of a successful experiment that eliminates or minimizes potential artefacts have been identified, and the specifically crystallographic components of these strategies have been implemented. The remaining computational challenge is to identify and extract time-independent structures, each corresponding to a distinct reaction intermediate, whose populations vary with time and give rise to the time-dependent X-ray diffraction data. The fourth dimension, time, has been added to the three spatial dimensions of crystallography; it can no longer be regarded as purely a static technique.


Anti-interferon-gamma antibodies in the treatment of autoimmune diseases.
Interferon (IFN)-gamma is an important immune regulator in normal immunity. When IFN gamma production is disturbed, various autoimmune diseases (ADs) can develop, in which we suggest that anti-IFN gamma could have a beneficial effect. Depending on the cell type in which IFN gamma synthesis is disturbed, different clinical manifestations may result. We have also proposed to remove tumor necrosis factor (TNF)-alpha, together with certain types of IFNs, to treat various ADs and AIDS, also an autoimmune condition. Anti-IFN gamma has been tested in several T-helper cell (Th1) ADs, including rheumatoid arthritis (RA), multiple sclerosis (MS), corneal transplant rejection, uveitis, Type I diabetes, schizophrenia (anti-IFN gamma and anti-TNF alpha), and various autoimmune skin diseases (alopecia areata, psoriasis vulgaris, vitiligo, pemphigus vulgaris and epidermolysis bullosa). A strong, sometimes striking, therapeutic response followed administration of anti-IFN gamma, indicating that it may be a promising therapy for Th1 ADs.


α3β4∗ Nicotinic Acetylcholine Receptors Strongly Modulate the Excitability of VIP Neurons in the Mouse Inferior Colliculus.
The inferior colliculus (IC), the midbrain hub of the central auditory system, receives extensive cholinergic input from the pontomesencephalic tegmentum. Activation of nicotinic acetylcholine receptors (nAChRs) in the IC can alter acoustic processing and enhance auditory task performance. However, how nAChRs affect the excitability of specific classes of IC neurons remains unknown. Recently, we identified vasoactive intestinal peptide (VIP) neurons as a distinct class of glutamatergic principal neurons in the IC. Here, in experiments using male and female mice, we show that cholinergic terminals are routinely located adjacent to the somas and dendrites of VIP neurons. Using whole-cell electrophysiology in brain slices, we found that acetylcholine drives surprisingly strong and long-lasting excitation and inward currents in VIP neurons. This excitation was unaffected by the muscarinic receptor antagonist atropine. Application of nAChR antagonists revealed that acetylcholine excites VIP neurons mainly via activation of α3β4∗ nAChRs, a nAChR subtype that is rare in the brain. Furthermore, we show that acetylcholine excites VIP neurons directly and does not require intermediate activation of presynaptic inputs that might express nAChRs. Lastly, we found that low frequency trains of acetylcholine puffs elicited temporal summation in VIP neurons, suggesting that in vivo-like patterns of cholinergic input can reshape activity for prolonged periods. These results reveal the first cellular mechanisms of nAChR regulation in the IC, identify a functional role for α3β4∗ nAChRs in the auditory system, and suggest that cholinergic input can potently influence auditory processing by increasing excitability in VIP neurons and their postsynaptic targets.


The Microtubule-Associated Innate Immune Sensor GEF-H1 Does Not Influence Mouse Norovirus Replication in Murine Macrophages.
Norovirus is an acute infection of the gastrointestinal tract causing rapid induction of vomiting and diarrhoea. The infection is sensed and controlled by the innate immune system, particularly by the RNA helicase MDA-5 and type I and III interferons (IFNs). We have observed that intracellular replication of murine norovirus (MNV) occurs in membranous clusters proximal to the microtubule organising centre, a localisation dependent on intact microtubules. Recently, it was shown that the host protein guanine nucleotide exchange factor-H1 (GEF-H1) is a microtubule-associated innate immune sensor that activates interferon Regulatory Factor 3 to induce the production of type I IFNs. Thus, we interrogated the potential role of GEF-H1 in controlling MNV infections. We observed that GEF-H1 was recruited to the MNV replication complex; however RNAi-mediated suppression of GEF-H1 did not outwardly affect replication. We furthered our studies to investigate the impact of GEF-H1 on MNV innate detection and observed that GEF-H1 did not contribute to type I IFN induction during MNV infection or influenza virus infection but did result in a small reduction of interferon⁻β (IFNβ) during West Nile virus infection. Intriguingly, we discovered an interaction of GEF-H1 with the viral MNV non-structural protein 3 (NS3), an interaction that altered the location of GEF-H1 within the cell and prevented the formation of GEF-H1-induced microtubule fibres. Thus, our results indicate that GEF-H1 does not contribute significantly to the innate immune sensing of MNV, although its function may be modulated via interaction with the viral NS3 protein.


Relactation: an effective intervention to promote exclusive breastfeeding.
Fifty mothers of hospitalized infants less than 4 months old with partial or complete lactation failure (LF) were randomly assigned to two groups of 25 each for relactation. Majority of enrolled mothers (86 per cent) had complete LF and 14 per cent had partial LF. Relactation was attempted in both the groups with motivation, support, and repeated suckling. Group II mothers, in addition, were given metoclopramide. The characteristics of mothers and babies, and socio-economic background were comparable in both groups (P > 0.05). The primary outcome variables measured included the time of appearance of first breast milk secretion (in complete LF), time for partial and complete relactation. Relactation attempt was successful in 49 mothers (98 per cent), with complete relactation in 46 (92 per cent) and only partial relactation in three mothers (6 per cent). Nipple confusion and frustration of the baby (4 per cent) in complete LF, small/retracted nipples (8 per cent), and sore/fissured nipples (4 per cent), which influenced the initiation of relactation, were overcome with drop and drip method, lact-aid support and proper positioning of the baby. All the outcome variables of the two groups were comparable (P > 0.05). The pattern of weight gain, the rate of reduction in the amount of top milk and subsequent weight gain in the follow-up was also comparable in both the groups (P > 0.05). Maternal factors like breast conditions, nutrition, parity, feeding practices in previous babies, lactation gap, and infants' initial refusal to suck at the breast did not influence the outcome variables as long as repeated suckling was ensured. We conclude that relactation is possible in most of the mothers without the help of lactagogues. Apart from educating and motivating the mother for exclusive breastfeeding during first 4-6 months, a strong professional support by a skilled health worker is needed to overcome the initial problems during relactation.


Reproductive Justice Beyond Borders: Global Feminist Solidarity in the Post-Roe Era.
The global impact of Dobbs v. Jackson Women's Health Organization and the backlash towards reproductive justice that it represents warrant a global feminist response informed by broad theoretical and geopolitical lenses. We consider how a solidaristic, transnational feminist movement might learn from Latin American feminist movements that have been successful in uniting broad coalitions in the fight for reproductive justice as situated within far-reaching political goals. The success of such a global movement must be decolonial and must contend with the fact that overlapping realities of global inequality, severe poverty, extractivism, and western-backed violence are fundamentally implicated in reproductive justice.


A molecular re-appraisal of taxa in the Sordariomycetidae and a new species of Rimaconus from New Zealand.
Several taxa that share similar ascomatal and ascospore characters occur in monotypic or small genera throughout the Sordariomycetidae with uncertain relationships based on their morphology. Taxa in the genera Duradens, Leptosporella, Linocarpon, and Rimaconus share similar morphologies of conical ascomata, carbonised peridia and elongate ascospores, while taxa in the genera Caudatispora, Erythromada and Lasiosphaeriella possess clusters of superficial, obovoid ascomata with variable ascospores. Phylogenetic analyses of 28S large-subunit nrDNA sequences were used to test the monophyly of these genera and provide estimates of their relationships within the Sordariomycetidae. Rimaconus coronatus is described as a new species from New Zealand; it clusters with the type species, R. jamaicensis. Leptosporella gregaria is illustrated and a description is provided for this previously published taxon that is the type species and only sequenced representative of the genus. Both of these genera occur in separate, well-supported clades among taxa that form unsupported groups near the Chaetosphaeriales and Helminthosphaeriaceae. Lasiosphaeriella and Linocarpon appear to be polyphyletic with species occurring in several clades throughout the subclass. Caudatispora and Erythromada represented by single specimens and two putative Duradens spp. have unclear affinities in the Sordariomycetidae.


Efficiency of systemic versus intralesional bone marrow-derived stem cells in regeneration of oral mucosa after induction of formocresol induced ulcers in dogs.
Bone marrow mesenchymal stem cells (BMSCs) are the key to regenerative wound healing. MSCs have spatial memory and respond to local environment. The goal of this study was to evaluate the use of systemic and intralesional transplantation of BMSCs for regeneration of oral mucosa in an in vivo dog model. Transplantation of undifferentiated green fluorescent protein (GFP)-labeled autologous BMSCs systemically, submucosally or vehicle (saline) was injected around the chemically induced oral ulcer in each group of 18 adult dogs. The healing process of the ulcer was monitored clinically and histopathologically. Gene expression of vascular endothelial growth factor (VEGF) and collagen genes was detected in biopsies from all ulcers. One way ANOVA was used to compare between means of the three groups. Results were considered significant at P < 0.05. Flow cytometric analysis of the MSCs at the passage 3 showed that these cells were negative for CD45 (2.39%). They expressed high levels of CD29 (98.34%). Frozen fluorescence microscopy of sections of the cell-treated oral tissue of all groups indicated that the GFP-transduced implanted cells were integrated within the transplanted tissues. The treatment resulted in dramatic wound edge activation and resurfacing of oral mucosa wound. Our results revealed that BMSCs may be labeled with (GFP), in order to know the distribution of these cells after administration, and suggest that intralesional administration is an appropriate procedure to achieve acceptable regeneration of the previously injured oral mucosa more than systemic route.


Skin cancer awareness in suburban employees: a Hispanic perspective.
Skin cancer is the most common form of cancer in the United States and accounts for an estimated 1 million new cases per year. In the white population, the rate of new melanomas diagnosed yearly has more than doubled from 5/100,000 in 1973 to 12/100,000 in 1990. The current lifetime risks for white persons are estimated to be 28% to 33% for basal cell carcinoma and 7% to 11% for squamous cell carcinoma. Skin cancer is more common in sun-sensitive, fair-skinned individuals but can occur in any ethnic group, especially with exposure in tropical or sunny climates. Little is known about awareness of skin cancer, risk perception, and performance of skin self-examination (SSE) by people with skin that rarely burns. Our purpose was to evaluate skin cancer awareness, perceptions of skin cancer risk, and performance of SSE in a Hispanic versus non-Hispanic white population with similar access to health care and promotion. One hundred forty people employed by a suburban city voluntarily attended a free skin cancer screening and completed a self-administered survey. Hispanic individuals reported decreased skin sensitivity and tendency to burn. Non-Hispanic individuals believed they were at greater than average risk for skin cancer, and most Hispanics believed they were at average or below average risk. None of the 27 Hispanics reported ever being taught SSE. More non-Hispanics (32%) had performed SSE within the last year than Hispanics (15%). Regular SSE was associated with a history of skin cancer. Multimedia messages usually link skin cancer warning signs and early detection strategies with having sun-sensitive skin. People without sun sensitivity did not perceive themselves as being at risk, did not learn the warning signs of skin cancer, and did not perform SSE. Awareness of melanoma and non-melanoma skin cancer and perception of risk among Hispanics were less than among non-Hispanics, which may contribute to presentation for care at an advanced stage.


Virulence of Pseudomonas syringae pv. tomato DC3000 Is Influenced by the Catabolite Repression Control Protein Crc.
Pseudomonas syringae infects diverse plant species and is widely used as a model system in the study of effector function and the molecular basis of plant diseases. Although the relationship between bacterial metabolism, nutrient acquisition, and virulence has attracted increasing attention in bacterial pathology, it is largely unexplored in P. syringae. The Crc (catabolite repression control) protein is a putative RNA-binding protein that regulates carbon metabolism as well as a number of other factors in the pseudomonads. Here, we show that deletion of crc increased bacterial swarming motility and biofilm formation. The crc mutant showed reduced growth and symptoms in Arabidopsis and tomato when compared with the wild-type strain. We have evidence that the crc mutant shows delayed hypersensitive response (HR) when infiltrated into Nicotiana benthamiana and tobacco. Interestingly, the crc mutant was more susceptible to hydrogen peroxide, suggesting that, in planta, the mutant may be sensitive to reactive oxygen species generated during pathogen-associated molecular pattern-triggered immunity (PTI). Indeed, HR was further delayed when PTI-induced tissues were challenged with the crc mutant. The crc mutant did not elicit an altered PTI response in plants compared with the wild-type strain. We conclude that Crc plays an important role in growth and survival during infection.


Genomic Stability and Genetic Defense Systems in Dolosigranulum pigrum, a Candidate Beneficial Bacterium from the Human Microbiome.
Dolosigranulum pigrum is positively associated with indicators of health in multiple epidemiological studies of human nasal microbiota. Knowledge of the basic biology of D. pigrum is a prerequisite for evaluating its potential for future therapeutic use; however, such data are very limited. To gain insight into D. pigrum's chromosomal structure, pangenome, and genomic stability, we compared the genomes of 28 D. pigrum strains that were collected across 20 years. Phylogenomic analysis showed closely related strains circulating over this period and closure of 19 genomes revealed highly conserved chromosomal synteny. Gene clusters involved in the mobilome and in defense against mobile genetic elements (MGEs) were enriched in the accessory genome versus the core genome. A systematic analysis for MGEs identified the first candidate D. pigrum prophage and insertion sequence. A systematic analysis for genetic elements that limit the spread of MGEs, including restriction modification (RM), CRISPR-Cas, and deity-named defense systems, revealed strain-level diversity in host defense systems that localized to specific genomic sites, including one RM system hot spot. Analysis of CRISPR spacers pointed to a wealth of MGEs against which D. pigrum defends itself. These results reveal a role for horizontal gene transfer and mobile genetic elements in strain diversification while highlighting that in D. pigrum this occurs within the context of a highly stable chromosomal organization protected by a variety of defense mechanisms. IMPORTANCE Dolosigranulum pigrum is a candidate beneficial bacterium with potential for future therapeutic use. This is based on its positive associations with characteristics of health in multiple studies of human nasal microbiota across the span of human life. For example, high levels of D. pigrum nasal colonization in adults predicts the absence of Staphylococcus aureus nasal colonization. Also, D. pigrum nasal colonization in young children is associated with healthy control groups in studies of middle ear infections. Our analysis of 28 genomes revealed a remarkable stability of D. pigrum strains colonizing people in the United States across a 20-year span. We subsequently identified factors that can influence this stability, including genomic stability, phage predators, the role of MGEs in strain-level variation, and defenses against MGEs. Finally, these D. pigrum strains also lacked predicted virulence factors. Overall, these findings add additional support to the potential for D. pigrum as a therapeutic bacterium.


Products of cells cultured from gliomas. VI. Immunofluorescent, morphometric, and ultrastructural characterization of two different cell types growing from explants of human gliomas.
Explants derived from human gliomas have been characterized with respect to their cellular outgrowth pattern after 1-22 weeks in culture. A mat of cells which were fibronectin (FN)-positive and glial fibrillary acidic protein (GFAP)-negative (hereafter designated FN+ cells) with a polygonal, flat morphology covered the growth substrate in a swirling pattern for a mean diameter of 9.2 mm around FN+ explants. FN+ cells showed ruffled plasmalemma, dilated rough endoplasmic reticulin (RDR), and extracellular filamentous strands. Rare desmosomes were compatible with at most minor leptomeningeal components or differentiation. FN+ cells predominated in six of seven cultures at passage 2, and their features were the same from various high-grade gliomas and gliosarcoma. Around other explants, elongated or stellate cells which were GFAP+ and FN- grew in a netlike pattern with little cell-to-cell contact. These GFAP+ cells surrounded explants at a mean diameter of 2 mm, substantially less than FN+ cells (P less than 0.005), and they grew more slowly than FN+ cells around explants. GFAP+ cells had an area/perimeter ratio which was less than that of FN+ cells. GFAP+ cells contained abundant intracellular filaments, rare desmosomes, and narrow RER cisternae. In mixed explants, GFAP+ cells often grew on top of FN+ cells. Individual cells which stained for both GFAP and FN were evident only from one glioma (8% doubly positive). Cells negative for both proteins resembled FN+ cells morphologically. Frozen sections of original glioma tissue showed FN+ vessel walls and GFAP+ parenchyma. Results are evidence for very early overgrowth of a preexistent FN+ cell type distinct from the GFAP+ parenchymal cell. The features of this distinct cell type are mesenchymal and resemble the proliferating vascular elements of gliomas in situ. The tendency for GFAP+ cells to grow on top of these FN+ cells suggests a feeder layer interaction. More knowledge of the origins and interactions of these two cell types may increase our understanding of the mechanism of antigenic changes in gliomas and may provide clues to improved therapeutic approaches.


The molecular-weight distribution of glycosaminoglycans.
1. A rapid and sensitive method for the accurate estimation of the molecular-weight distribution of keratan sulphate and chondroitin sulphate isolated from adult bovine nasal septum and intervertebral disc is described. The method utilizes gel chromatography of reductively labelled glycosaminoglycan and end-group estimation of number-average molecular weight for each fraction across the peak of eluted glycosaminoglycan. 2. Chain-length distribution data obtained by this procedure are used to evaluate mechanisms of chondroitin sulphate biosynthesis.


Detrimental effects of late referral for dialysis.
The potential benefits of early referral to a nephrologist of chronic uremics include slowing the rate of decline associated with progressive renal insufficiency and facilitating efficient entry into dialysis programs. Late referral is associated with complications that are uncommon among early referrals. This study aim to evaluate the pattern of referral among chronic uremics with a view to identifying associated problems. All the patients seen with chronic renal failure during a five-year period in the dialysis unit of the hospital had their medical records reviewed. Information was extracted on the clinical data and laboratory records. Ninety patients were seen in the dialysis unit of the hospital during this period. The mean age was 38.3+ 1.6 years (range 10-69 years) with a peak incidence in the second decade. Most patients were referred late from the private medical clinics 34(37.8 % ) and the general hospitals 32(35.6 % ). Seventy (77.8 % ) patients were initially diagnosed as chronic uremics, six months prior to referral. The aim of referring in most cases was persistent uremic symptoms and necessity for dialysis. Only 14(15.6 %) patients presented without complications. The commonest were pulmonary edema, (28.9 % ) biventricular failure (22.2 % ) and hypertension (20 % ). The majority of the patients were referred late to nephrologists and presented with complications. Chronic Renal Failure patients should be referred early to nephrologists so as to prevent/reduce morbidity and mortality.


Feline acromegaly: a review of the syndrome.
Acromegaly is characterized by chronic excessive growth hormone (GH) secretion by the pituitary gland. Feline acromegaly is most commonly caused by a functional pituitary tumor. Definitive diagnosis can be difficult because of the gradual disease onset, subtle clinical signs, unavailability of relevant laboratory tests, and client financial investment. The most significant clinical finding of acromegaly is the presence of insulin-resistant diabetes mellitus. Diagnosis is currently based upon brain imaging and measurement of serum GH and/or insulin-like growth factor-1 concentrations. Definitive treatment in cats is not well described, but radiation therapy appears promising.


A fairy tale of modern insulin therapy in type 1 diabetes.
In type 1 diabetes, flexible, intensive insulin management improves not only glycemic control but also dietary freedom and treatment satisfaction. Such flexibility has been made possible with the new insulin analogues (as part of a basal-bolus regime) and is now gaining wide applicability, especially among children and adolescents. This approach requires appropriate individualized patient education. Especially for adolescents, the clinician should be able to guarantee insightful participation in direct response to their attitudes, wishes and needs. This patient-and-doctor collaboration is an ever-challenging duty and has the potential to change the future of the individual diabetic patient.


Drug-induced Bile Duct Injury - A Short Review.
The liver represents the major site of drug metabolism, i.e. the key organ in the processes of detoxification and elimination of drugs from the organism. It is therefore often affected by toxic metabolites and suffers sometimes fatal consequences. The spectrum of pathologies differs by the cell type primarily damaged and the group of the cholangiopathies includes those conditions affecting the bile duct epithelium or the cholangiocytes. They can range from transient cholestasis to vanishing bile duct syndrome and sclerosing cholangitis, both leading eventually to the development of biliary fibrosis and cirrhosis. In this review article, we focus on the etiology, predisposing factors, clinical manifestations, and histopathological characteristics of bile duct injury as a consequence of drug treatment and discuss separately the different bile duct pathologies.


Characterization of estrogen binding sites in the liver of the newt Pleurodeles waltl (Urodela:Amphibia): demonstration of a sex-linked heterogeneity.
Saturation analysis over a wide range of [3H]estradiol-17 beta concentrations (1-40 nM) in cytosols prepared from liver of the newt Pleurodeles waltl of both sexes revealed a sex-linked heterogeneity of the estradiol-17 beta binding sites. In females, one type of binding site has been identified as a classical receptor. It exhibited a high affinity for estradiol-17 beta (Kd = 9 X 10(-9) M), had a high specificity for estrogenic compounds and was stabilized by monothioglycerol. In males, in addition to the receptor found in females, a second estrogen binding component was detected, not observed in female cytosols. It exhibited a Kd of 4.8 X 10(-8) M for estradiol 17 beta, higher capacity and displayed the same highly specific estrogen binding as does the estrogen receptor. It was affected by monothioglycerol and its binding was found to be significantly increased on cytosol dilution, as well as by estrogen-treatment.


Sleep quality change after upper airway surgery in obstructive sleep apnea: Electrocardiogram-based cardiopulmonary coupling analysis.
To test the effect of upper airway surgery on sleep quality in adults with obstructive sleep apnea (OSA) and the potential usefulness of electrocardiogram (ECG)-based cardiopulmonary coupling (CPC) analysis as metrics of sleep quality. Retrospective outcome research. A total of 62 consecutive adult patients with OSA, consisting of 36 with successful and 26 with unsuccessful outcomes, were included in the study. Mean age was 37.7 ± 8.9 years, and body mass index (BMI, kg/m(2) ) was 26.9 ± 2.3. We compared clinical characteristics (age, BMI, and Epworth Sleepiness Scale [ESS]), sleep (sleep efficiency, stage non-rapid eye movement [N]1, N2, N3, rapid eye movement, and arousal index [ArI]), respiratory (apnea index [AI], apnea-hypopnea index [AHI], and minimum arterial oxygen saturation [SaO2 ]), and CPC (high-frequency coupling [HFC], low frequency coupling [LFC], very-low-frequency coupling, and elevated low-frequency coupling [e-LFC]) parameters between the success and nonsuccess groups before and after surgery. Surgical success was defined when the postoperative AHI was both <20 per hour and 50% of the preoperative value. Sleep quality measured by CPC analysis improved significantly (HFC, P = .001; LFC, P = .002; e-LFC, P = .003), along with parallel reduction in ESS, respiratory parameters (AHI, AI, minimum SaO2 ), and sleep fragmentation (ArI) in the group with surgical success after upper airway surgery. Successful upper airway surgery can improve objective sleep quality in adult patients with OSA. CPC metrics of sleep quality are potentially useful to monitor therapeutic responses during long-term postoperative follow-up, as the ECG-based analysis is available as a standalone option outside laboratory polysomnography.


Levels of anti-albumin autoantibodies in acute and chronic hepatitis B virus infection.
Levels of anti-albumin autoantibodies (AAA) of the IgG class were determined by ELISA in sera of patients with acute and chronic hepatitis B virus (HBV) infection. Mean AAA levels were higher than normal in both acute and chronic hepatitis patients. AAA levels were higher than the upper normal limits in practically all patients with acute self-resolving hepatitis, and decreased to normal levels when the patients recovered. Enhanced IgG AAA levels were observed in many patients with chronic hepatitis and serological markers of HBV replication. Elevated AAA levels were not associated with either more elevated transaminase levels or more severe histological forms of chronic hepatitis. The results of this study suggest that the interaction of albumin with HBV determinants is involved in AAA elevations, probably by mediating an increase in albumin immunogenicity. Moreover, the fall in AAA levels in the recovery phase of acute hepatitis, the coexistence of elevated levels with HBeAg and HBV-DNA, and the lack of correlation between AAA levels and different evolutive forms of chronic hepatitis, seem to exclude AAA from playing a relevant role, be it protective or damaging, during HBV infection.


Incidence and distribution of UroSEEK gene panel in a multi-institutional cohort of bladder urothelial carcinoma.
Noninvasive approaches for early detection of bladder cancer are actively being investigated. We recently developed a urine- based molecular assay for the detection and surveillance of bladder neoplasms (UroSEEK). UroSEEK is designed to detect alterations in 11 genes that include most common genetic alterations in bladder cancer. In this study, we analyzed 527 cases, including 373 noninvasive and 154 invasive urothelial carcinomas of bladder from transurethral resections or cystectomies performed at four institutions (1991-2016). Two different mutational analysis assays of a representative tumor area were performed: first, a singleplex PCR assay for evaluation of the TERT promoter region (TERTSeqS) and second, a multiplex PCR assay using primers designed to amplify regions of interest of 10 (FGFR3, PIK3CA, TP53, HRAS, KRAS, ERBB2, CDKN2A, MET, MLL, and VHL) genes (UroSeqS). Overall, 92% of all bladder tumors were positive for at least one genetic alteration in the UroSEEK panel. We found TERT promoter mutations in 77% of low-grade noninvasive papillary carcinomas, with a relatively lower incidence of 65% in high-grade noninvasive papillary carcinomas and carcinomas in situ; p = 0.017. Seventy-two percent of pT1 and 63% of muscle-invasive bladder tumors harbored TERT promoter mutations with g.1295228C>T alteration being the most common in all groups. FGFR3 and PIK3CA mutations were more frequent in low-grade noninvasive papillary carcinomas compared with high-grade noninvasive papillary carcinomas and carcinomas in situ (p < 0.0001), while the opposite was true for TP53 (p < 0.0001). Significantly higher rates of TP53 and CDKN2A mutation rates (p = 0.005 and 0.035, respectively) were encountered in muscle-invasive bladder tumors compared with those of pT1 stage. The overwhelming majority of all investigated tumors showed at least one mutation among UroSEEK assay genes, confirming the comprehensive coverage of the panel and supporting its potential utility as a noninvasive urine-based assay.


The immediate effect of a soft knee brace on dynamic knee instability in persons with knee osteoarthritis.
Wearing a soft knee brace has been shown to reduce self-reported knee instability in persons with knee OA. There is a need to assess whether a soft knee brace has a beneficial effect on objectively assessed dynamic knee instability as well. The aims of the study were to evaluate the effect of a soft knee brace on objectively assessed dynamic knee instability and to assess the difference in effect between a non-tight and a tight soft knee brace in persons with knee OA. Thirty-eight persons with knee OA and self-reported knee instability participated in a laboratory study. A within-subject design was used comparing no brace vs brace and comparing a non-tight vs a tight brace. The primary outcome measure was dynamic knee instability, expressed by the perturbation response (PR). The PR reflects deviation in the mean knee varus-valgus angle during level walking after a controlled mechanical perturbation. Linear mixed-effect model analysis was used to evaluate the effect of a brace on dynamic knee instability. Wearing a brace significantly reduced the PR compared with not wearing a brace (B = -0.16, P = 0.01). There was no difference between a non-tight and a tight brace (B = -0.03, P = 0.60). This study is the first to report that wearing a soft knee brace reduces objectively assessed dynamic knee instability in persons with knee OA. Wearing a soft brace results in an objective improvement of knee instability beyond subjectively reported improvement. Nederlands Trial register (trialregister.nl) NTR6363.


Modulation of elastin expression and cell proliferation in vascular smooth muscle cells in vitro.
Elastin expression in cultured vascular smooth muscle cell (VSMC) was found to be enhanced by potent inhibitors of VSMC proliferation including minoxidil, heparin and retinoic acid. By contrast, elastin expression was declined by potent stimulators of VSMC proliferation like epidermal growth factor, high K+, angiotension II and phorbol ester. To elucidate the relationship between elastin expression and cell proliferation, the elastin expression in the different cell growth states brought by cell-synchronizing culture or suspension culture, a culture system independent of potent modulators of VSMC proliferation. Elastin was found to be expressed maximally at G0 and minimally at G2/M phases, suggesting that cell growth state regulates elastin expression in VSMC culture. Synthetic elastin peptide VPGVG or its polymeric form (VPGVG)n enhanced VSMC proliferation, which resulted in the reduction of elastin expression. The results suggests that elastin fragment regulates VSMC proliferation. These correlation between elastin expression and cell growth state may play an essential role in elastin metabolism under the normal and diseased conditions.


Glutathione S-transferase polymorphisms and risk of differentiated thyroid carcinomas: a case-control analysis.
To determine the association between glutathione S-transferase (GST) polymorphisms and the risk of differentiated thyroid carcinoma (DTC) and benign thyroid tumors. Case-control study. Tertiary care cancer center. Two hundred one patients with DTC, 103 patients with benign thyroid tumors, and 680 cancer-free control subjects. Results of a polymerase chain reaction-based assay for genotyping. A multivariate logistic regression analysis was performed with adjustment for age, sex, ethnicity, tobacco use, and alcohol use. The patients with DTC were younger, more likely to be female and nonwhite, and less likely to smoke or consume alcohol than the controls. Overall, 55.2% of the DTC cases and 52.6% of the controls were null for the gene for GST-mu1 (GSTM1) (P = .52), and 25.4% of the DTC subjects and 20.6% of the controls were null for the GST-theta1 gene (GSTT1) (P = .15). However, 15.9% of the DTC cases but only 9.4% of the controls were null for both genes (P = .009). In addition, the results of the adjusted multivariate regression analysis suggested that having both null genotypes was associated with an increased risk for DTC (odds ratio [OR], 2.1 [95% confidence interval, 1.3-3.5; P = .003]). This was particularly true for women (OR, 2.5), current smokers (OR, 3.6), and nonwhites (OR, 5.6). A similar analysis demonstrated a nonsignificant association between these genotypes and benign thyroid tumors (OR, 1.5 [95% confidence interval, 0.7-3.0; P=.30). Our results suggest that the simultaneous presence of the GSTM1- and GSTT1-null genotypes is a susceptibility factor for DTC. Such knowledge may ultimately help refine cancer prevention efforts; however, larger studies are needed to verify these findings.