The role of radiotherapy in rectal cancer.
In Europe, short-term radiotherapy is increasingly used in the primary management of rectal cancer. In the United States, postoperative chemoradiotherapy is the standard treatment of choice. The rationale and indications for radiotherapy and possible combinations with chemotherapy are discussed and an overview of all of the randomised trials containing radiotherapy as one randomisation arm is given. Three major indications for radiotherapy can be identified: the reduction of local recurrences in mobile rectal cancer, downstaging of the tumour in primary irresectable tumours and downsizing of low-lying tumours in attempts to more frequently perform a sphincter-saving procedure. For reduction of local recurrences, radiotherapy can be given either pre- or postoperatively, although preoperative therapy is more dose-efficient. Short-term preoperative radiotherapy reduces the number of recurrences and improves survival. Improved survival is also reported after postoperative radiotherapy in combination with chemotherapy, however, the relevance of the radiotherapy component is discussed. Although the debate about radiotherapy is still ongoing, we strongly believe that the results demonstrate that short-term preoperative radiotherapy is the treatment of choice for resectable rectal cancer.


Glycation associated skin autofluorescence and skin elasticity are related to chronological age and body mass index of healthy subjects.
Glycation is the non-enzymatic reaction between reducing sugars and proteins that leads to the formation of advanced glycation end products (AGEs). In vivo skin autofluorescence (lambda(ex)/lambda(em)=370/440 nm) was used as a non-invasive clinical tool to study skin AGE accumulation in healthy panellists. Using multiple linear regression analysis, it was shown that for panellists below the age of 40, glycation associated in vivo skin fluorescence intensity increased as a function of chronological age and body mass index (BMI). Above the age of 40, the fluorescence was associated to age but not to BMI, suggesting that the effect of age became dominant over BMI. Since the accumulation of AGEs is expected to affect the biomechanical properties of the skin, in vivo skin elasticity data were gathered on a second panel. It was found that skin elasticity depended on age and BMI in a similar fashion as to what we observed for the skin fluorescence data. It is hypothesised that skin AGE accumulation contributes to the loss of skin elasticity in aged and/or overweight people.


Accumulation and depuration of the synthetic antioxidant ethoxyquin in the muscle of Atlantic salmon (Salmo salar L.).
The biological fate of the fish feed additive, ethoxyquin (EQ) was examined in the muscle of Atlantic salmon during 12 weeks of feeding followed by a 2 weeks depuration period. Parent EQ (1,2-dihydro-6-ethoxy-2,2,4-trimethylquinoline), quinone imine (2,6-dihydro-2,2,4-trimethyl-6-quinolone), de-ethylated EQ (6-hydroxy-2,2,4-trimethyl-1,2-dihydroquinoline) and EQDM (EQ dimer or 1,8'-di(1,2-dihydro-6-ethoxy-2,2,4-trimethyl-quinoline) were found to be the ubiquitous metabolites of dietary EQ, with EQDM as a main metabolite. A rapid decrease in the level of EQ (2.4 days of half-life) was balanced by an increase in EQDM, giving an unchanged net sum following 2 weeks of depuration. The mandatory 14 days depuration period prior to slaughtering of farmed salmon in Norway was not sufficient for complete elimination of EQ-derived residuals. Post depuration, EQDM accounted for 99% of sum of the two compounds in all treatment groups; possible toxicological effects of EQDM are not known. The individual concentrations of EQ and EQDM and their sum are dependent on EQ level in the feed, consequently, their residual concentrations may be controlled. The theoretical amount of EQ and EQDM consumed in one meal of farmed salmon would be under the recommended ADI, provided that the fish were raised on feed with no more than 150 mg EQ/kg feed, which is the EU maximum limit for EQ in fish feed.


Proteomic Detection and Validation of Translated Small Open Reading Frames.
Small open reading frames (smORFs) encode previously unannotated polypeptides or short proteins that regulate translation in cis (eukaryotes) and/or are independently functional (prokaryotes and eukaryotes). Ongoing efforts for complete annotation and functional characterization of smORF-encoded proteins have yielded novel regulators and therapeutic targets. However, because they are excluded from protein databases, initiate at non-AUG start codons, and produce few unique tryptic peptides, unannotated small proteins cannot be detected with standard proteomic methods. Here,, we outline a procedure for mass spectrometry-based detection of translated smORFs in cultured human cells from protein extraction, digestion, and LC-MS/MS, to database preparation and data analysis. Following proteomic detection, translation from a unique smORF may be validated via siRNA-based silencing or overexpression and epitope tagging. This is necessary to unambiguously assign a peptide to a smORF within a specific transcript isoform or genomic locus. Provided that sufficient starting material is available, this workflow can be applied to any cell type/organism and adjusted to study specific (patho)physiological contexts including, but not limited to, development, stress, and disease. © 2019 by John Wiley & Sons, Inc. Basic Protocol 1: Protein extraction, size selection, and trypsin digestion Alternate Protocol 1: In-solution C8 column size selection Support Protocol 1: Chloroform/methanol precipitation Support Protocol 2: Reduction, alkylation, and in-solution protease digestion Support Protocol 3: Peptide de-salting Basic Protocol 2: Two-dimensional LC-MS/MS with ERLIC fractionation Basic Protocol 3: Transcriptomic database construction Alternate Protocol 2: Transcriptomics database generation with gffread Basic Protocol 4: Non-annotated peptide identification from LC-MS/MS data Basic Protocol 5: Validation using isotopically labeled synthetic peptide standards and siRNA Basic Protocol 6: Transcript validation using transient overexpression.


Isolation and characterization of microsatellite markers for the Korean rockfish, Sebastes schlegeli.
The Korean rockfish (Sebastes schlegeli) is an important commercial fish that is widely used in aquaculture. We isolated and characterized 18 polymorphic microsatellite loci from the Korean rockfish using a (GT)(13)-enriched genomic library. Polymorphism was assessed in 48 individuals from a single population collected from the northern coastal waters of the Yellow Sea. The observed and expected heterozygosities ranged from 0.0244 to 0.7660 (mean 0.4194) and 0.0244 to 0.8758 (mean 0.5002), respectively. Polymorphism at these loci indicated from two to 15 alleles (mean 5.7); 14 of 18 loci conformed to Hardy-Weinberg equilibrium. These markers should be useful for management and conservation studies of this species.


[Value of silver staining technics for the study of acrocentric ring chromosomes and supernumerary microchromosomes].
The silver staining techniques was used to study two cases of ring chromosomes (ring chromosome 15 and ring chromosome 22) and two cases of small extra chromosomes. This technique allows identification of the breakpoints and provides some information about the origin of small extra chromosomes.


Validity of Pinckers' 100-Hue version of the Panel D-15.
Fifteen normal and 18 red-green-deficient subjects were tested with the Farnsworth Dichotomous Test of color vision (Panel D-15) and with an alternative version of the dichotomous test consisting of selected color chips from the Farnsworth-Munsell 100-Hue Test. Results indicate that the 100-Hue version of the dichotomous test is equivalent to the standard Panel D-15 with respect to detecting and differentiating among the more severe red-green deficiencies. These results confirm A. Pinckers' previous suggestion that the 100-Hue Test contains the components necessary for a Farnsworth Panel D-15.


Thrombopoietin receptor agonists and rituximab for treatment of pediatric immune thrombocytopenia: A systematic review and meta-analysis of prospective clinical trials.
Children with immune thrombocytopenia (ITP) may require second-line ITP therapies. The high remission rate in pediatric patients, need for extended-duration use of thrombopoietin receptor agonists (TPO-RAs), drug adherence, potential side effects, monitoring, and cost effectiveness are factors that should be considered in decision-making about second-line therapies. Rituximab (RTX) has been used off-label for years to treat ITP but there are limited studies about its efficacy and safety in children. To date, no studies have directly compared TPO-RAs with RTX for the treatment of childhood ITP. This systematic review analyzed the overall platelet response, durability of treatment effect, and safety for RTX use in comparison to TPO-RAs in pediatric ITP. MEDLINE/PubMed, EMBASE, Cochrane Library, and Web of Science databases were searched through December 2020 and meta-analysis was conducted using proportions of success/failure for each intervention in the selected studies. The proportion of participants achieving the primary endpoint of a platelet response above 50,000 was similar for TPO-RAs (proportion = 0.71, 95% CI: 0.63-0.78) and RTX (proportion = 0.68, 95% CI: 0.53-0.82). However, considerable variation was found between the two groups with regards to the sustainability of the response and other secondary outcomes such as need for rescue and adverse events. RTX was associated with higher rates of rescue therapy. In this analysis of prospective pediatric ITP studies, RTX and TPO-RAs had similar rates of overall platelet response but differed in other important measures. Prospective comparative studies are needed to better characterize second-line treatments for pediatric ITP.


Oxygen radical production and trapping in subjects with previous Yersinia infection.
Previous studies indicate that oxygen radical production by 1-min activation of N-formyl-methionyl-leucyl-phenylalanine-induced chemiluminescence (CL) by whole blood is increased in subjects with previous Yersinia arthritis (YA). This finding is confirmed in the present study and extended further by demonstrating that subjects with previous YA show increased oxygen radical generation and enhanced release of myeloperoxidase from neutrophils and increased CL activity by serum, all factors that can contribute to an increase in the initial activation of CL. Initial activation was not increased in subjects who had had Yersinia enteritis without arthritis; however, intracellular oxygen radical production by purified neutrophils was significantly increased, suggesting that the cells had been primed in vivo. The water-soluble antioxidants of plasma samples tested by a peroxyl radical-trapping assay were much the same in subjects with previous YA, in subjects with previous Yersinia enteritis without arthritis, and in healthy subjects. The results suggest that aberrations in neutrophil oxygen radical production play a role in the pathogenesis of YA.


PI-103 sensitizes acute myeloid leukemia stem cells to daunorubicin-induced cytotoxicity.
To date, acute myeloid leukemia (AML) shows very poor outcome for conventional chemotherapy. Leukemia stem cells (LSCs) are insensitive to conventional chemotherapeutic drugs and play a central role in the pathogenesis of AML. Failure to effectively ablate these cells may lead to AML relapse following chemotherapy. Phosphatidylinositol 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) signaling pathway is constructively activated in LSCs. This pathway can be inhibited by PI-103, a novel synthesized molecule of the pyridofuropyrimidine class, resulting in the apoptosis of LSCs. Therefore, we investigate the influences of PI-103 in combination with daunorubicin (DNR) on the LSCs. Our data indicate that PI-103 synergistically sensitizes LSCs to DNR-induced cytotoxicity. In addition, the PI-103/DNR co-treatment can induce significant apoptosis in LSCs, but sparing hematopoietic stem cells. The synergistic effect and the LSCs-specific apoptosis mechanism may be associated with the inhibition of PI3K/Akt/mTOR signaling pathway. Our results suggest that PI-103 in combination with DNR may be a potent and less toxic therapy for targeting LSCs and deserve further preclinical and clinical studies in the treatment of AML.


Burns to the elderly: a reappraisal.
Admissions to the McIndoe burns unit over the period 1975 to 1984 were reviewed and 123 patients over the age of sixty-five were studied in detail. Comparison with standardised mortality curves showed no difference in survival rates, but review of individual cases suggests that regular examination of elderly patients could shorten admission times and reduce the mortality by encouraging prompt identification and treatment of medical complications. Patients with burns of greater than 5% of surface area should be resuscitated with intravenous fluids as this age group is very susceptible to hypotensive renal damage. Excision of burns on or before the fifth day resulted in longer admissions and increased the number of procedures per patient. Disagreement in the recent literature on this point suggests the need for a randomised, controlled trial of early excision in the elderly patient.


Comparison of the Cerebral Performance Category score and the Health Utilities Index for survivors of cardiac arrest.
The Cerebral Performance Category score is an easy to use but unvalidated measure of functional outcome after cardiac arrest. We evaluate the comparability of results from the Cerebral Performance Category scale versus those of the validated but more complex Health Utilities Index scale for health-related quality of life. This prospective substudy of the Ontario Prehospital Advanced Life Support (OPALS) Study included adult out-of-hospital cardiac arrest patients treated in 20 cities. This prospective cohort study included all survivors of out-of-hospital adult cardiac arrest enrolled in phase II (rapid basic life support with defibrillation) and phase III (advanced life support) of the OPALS Study, as well as the intervening run-in phase. Survivors were interviewed at 12 months for Cerebral Performance Category Score and the Health Utilities Index Mark 3 (Health Utilities Index). Of 8,196 eligible out-of-hospital cardiac arrest patients between 1995 and 2002, 418 (5.1%) survived to discharge, and 305 (3.7%) completed the Health Utilities Index interview and had Cerebral Performance Category scored at 12 months. The 305 patients had the following data: mean age 63.9 years; male 78.0%; paramedic-witnessed arrest 25.6%; bystander cardiopulmonary resuscitation 32.1%; initial rhythm ventricular fibrillation/ventricular tachycardia 86.9%, Cerebral Performance Category 1 267, Cerebral Performance Category 2 26, Cerebral Performance Category 3 12. Overall, the median scores (interquartile range) were Cerebral Performance Category 1 (1 to 1) and Health Utilities Index 0.84 (0.61 to 0.97). The Cerebral Performance Category score ruled out good quality of life (Health Utilities Index >0.80), with a sensitivity of 100% (95% confidence interval [CI] 98% to 100%) and specificity 27.1% (95% CI 20% to 35%); thus, when the Cerebral Performance Category score was 2 or 3, it was unlikely that the Health Utilities Index score would be good. The Cerebral Performance Category score had sensitivity 55.6% (95% CI 42% to 67%) and specificity 96.8% (95% CI 94% to 98%) for predicting poor quality of life (Health Utilities Index >0.40); ie, when Cerebral Performance Category was 1, it was highly unlikely that the Health Utilities Index score would be poor. The weighted kappa was 0.39 and the interclass correlation was 0.51. This represents the largest study yet conducted of the performance of the Cerebral Performance Category score in 1-year survivors of out-of-hospital cardiac arrest. Overall, the Cerebral Performance Category score classified patients well for their quality of life, ruling out a good Health Utilities Index score with high sensitivity and ruling in poor Health Utilities Index score with high specificity. The Cerebral Performance Category is an important tool in that it indicates broad functional outcome categories that are useful for a number of key clinical and research applications but should not be considered a substitute for the Health Utilities Index.


Novel textile dye obtained through transformation of 2-amino-3-methoxybenzoic acid by free and immobilised laccase from a Pleurotus ostreatus strain.
Transformation of 2-amino-3-methoxybenzoic acid into novel and eco-friendly orange dye (N15) was performed using native and immobilised laccase (LAC) from Pleurotus ostreatus strain. A several parameters affecting laccase-mediated transformation efficiency included the selection of type and pH value of buffer, reaction temperature, substrate and laccase concentration as well as the type of carrier and LAC storage conditions were evaluated. The optimal conditions for N15 dye synthesis were 40 mM sodium-tartrate buffer pH 5.5 containing 3 mM of the substrate, efficiently transformed by 2 U of free laccase per 1 mmol of the substrate. Laccase was immobilised on porous Purolite® carriers, which had never been tested as a support for oxidoreductases. Immobilised laccase, characterised by a high immobilisation yield, was obtained by adsorption of laccase on a porous acrylic carrier with octadecyl groups (C18) incubated in optimum conditions of 40 mM phosphate buffer pH 7.0 containing 1 mg of laccase per 1 g of the carrier (wet mass). The immobilised LAC showed the highest storage stability for 21 days and higher thermostability at 40 ℃ and 60 ℃ in comparison to its native form. The N15 dye showed good dyeing properties towards natural fibres, and the dyed fibre demonstrated resistance to different physicochemical factors during use, which was confirmed by commercial quality tests. The N15 dye is a phenazine, i.e. a heterogenic compound containing amino-, methoxy-, and three carboxyl functional groups with the molecular weight of approximately 449.37 U.


Medical Student Presyncope and Syncope in the Operating Room: A Mixed Methods Analysis.
Medical students often feel faint (presyncopal) in the operating room (OR). Despite mandatory surgical rotations in clerkship, there is little formal training and acclimatization to the operating suite in the first 2 years of medical school. This study aimed to assess presyncope and syncope in the OR in first, second, third, and fourth year medical students at a large Canadian academic center. Following an extensive literature review, we developed a mixed methods survey de novo to assess medical students' experience in the OR and determine the frequency of presyncopal and syncopal events. A total of 180 students (106 females, 59%), evenly distributed among all 4 years of medical school, responded to the survey (response rate 40%, n = 180/454). In total, 75 students (42%) had experienced presyncope, and 10 students (6%) had experienced syncope in the OR. Female medical students were more likely to experience both presyncope (p < 0.001) and syncope (p = 0.011) relative to their male colleagues. They were also more likely to report that these experiences had an effect on their attitude toward the OR (p < 0.001) and their career choice (p < 0.001). Half of respondents believed that the undergraduate medical preclerkship curriculum did not provide adequate exposure to the OR. Students consistently expressed the desire for more preclinical OR exposure and formal instruction concerning OR etiquette. Only 28% of students reported receiving information on how to avoid syncope, and their classmates were the number one source (59%) for this advice. The most commonly employed preventative measures were eating before the OR and staying well hydrated. Presyncope had a negative effect on attitudes toward the OR in 28% of students. Although true loss of consciousness (syncope) among medical students in the OR is uncommon, presyncope is a highly prevalent phenomenon. It is most prevalent in female students, and may have a negative effect on overall student well-being. All students may benefit from normalization of this very common experience by staff surgeons or residents. Formal instruction regarding the common symptoms, triggers, and methods for preventing presyncope and syncope is an essential supplement to the current preclinical medical curriculum.


Progressive changes in hepatoma cells stably transfected with hepatitis B virus X gene.
The aim of this study is to investigate the molecular mechanism of hepatocellular carcinoma (HCC) development induced by hepatitis B virus X protein (HBx). We previously established a H7402-X cell line that constitutively expresses HBx protein. In the present study, H7402-X gene expression profiles and proteins were examined using cDNA microarrays and Western blot analysis. Apoptosis was induced by adriamycin in H7402-X cells. The transcriptional activities of NF-kappaB and AP-1 were examined using a luciferase reporter gene. The DNA expression profiles identified candidate genes showing aberrant expression in cells overexpressing HBx. Western blot analysis showed that cyclin D, cyclin E, survivin, Bcl-2, and PCNA were up-regulated, whereas p27 was down-regulated in H7402-X cells. Treatment with RNAi targeting HBx mRNA led to the down-regulation of these genes. H7402-X cells were resistant to adriamycin-induced apoptosis. Luciferase reporter gene analysis revealed that HBx induces the transcriptional activities of NF-kappaB and AP-1. Our data provide additional insight into cellular targets of HBx, which allows a better understanding of HBx function and the progressive changes during HBx-mediated hepatocarcinogenesis.


Cesium and rubidium salts: effects on voluntary intake of ethanol by the rat.
The effects of RbCl and CsCl on voluntary intake of ethanol solution by rats preferring ethanol solution 5% (w/w) over water as the drinking fluid was studied as a function of the dose given and the vehicle injected. Administration of RbCl or CsCl, 0.5 mEq/kg/day or 1.5 mEq/kg/day for three consecutive days, did not alter amounts of ethanol consumed. Repeated administration of RbCl or CsCl, 3.0 mEq/kg/day for three days, produced some moderate reduction in ethanol consumption. Simultaneous injection of RbCl (1.5 mEq/kg) and CsCl (1.5 mEq/kg) resulted in greater and profound lasting decrease in ethanol drinking. The later treatment did not alter specific activities of rat liver alcohol- and aldehyde dehydrogenase from saline treated controls. In general, dissolving the chloride salt of the alkali metals in saline resulted in greater effects on ethanol drinking than that determined after identical dose injected with water as the vehicle. The possible mechanism(s) underlying the effects of alkali metal salts used are suggested.


Testing three pathways to substance use and delinquency among low-income African American adolescents.
Mounting literature suggests that parental monitoring, risky peer norms, and future orientation correlate with illicit drug use and delinquency. However, few studies have investigated these constructs simultaneously in a single statistical model with low income African American youth. This study examined parental monitoring, peer norms and future orientation as primary pathways to drug use and delinquent behaviors in a large sample of African American urban adolescents. A path model tested direct paths from peer norms, parental monitoring, and future orientation to drug use and delinquency outcomes after adjusting for potential confounders such as age, socioeconomic, and sexual orientation in a sample of 541 African American youth. Greater scores on measures of risky peer norms were associated with heightened risk of delinquency with an effect size that was twice in magnitude compared to the protective effects of future orientation. Regarding substance use, greater perceived risky peer norms correlated with the increased likelihood of substance use with a standardized effect size 3.33 times in magnitude compared to the protective effects of parental monitoring. Findings from this study suggest that interventions targeting risky peer norms among adolescent African American youth may correlate with a greater impact on reductions in substance use and delinquency than exclusively targeting parental monitoring or future orientation.


Congenital syphilis in aborted second trimester fetus: diagnosis by histological study.
A case of congenital syphilis in an aborted fetus in the second trimester of pregnancy was documented by demonstration of spirochaetes in the fetal liver, despite the fact that diagnostic studies had not been performed on the mother. Suspicion was aroused by the gross and microscopic appearance of the placenta, which appeared identical with the described in proven cases of congenital syphilis. Although these changes have been alleged to be non-specific, the rarity with which such well-developed lesions are encountered in placentas in other circumstances dictates the necessity for further study when they are observed.


The role of multiple attachments in intergenerational transmission of child sexual abuse among male victims.
According to Finkelhor (1984), difficulties child sexual abusers have in establishing adaptive adult relationships are a consequence of attachment problems with parents. Research shows that insecure attachment is associated with both the experience of child sexual abuse (CSA) as a victim and perpetration of CSA as an adult. Attachment may thus be a key factor in intergenerational transmission (IT) of CSA. The study aims to examine the direct, interactive, and mediated effects linking attachment in different relationships (mother, father, romantic partner) to IT of CSA among male victims. Thirty-two abusing and 27 non-abusing male CSA survivors were recruited, respectively, in a prison and via networks of former foster children or CSA victims. All participants completed the Attachment Multiple Model Interview to assess attachment along four dimensions (security, deactivation, hyperactivation, and disorganization) in the relationship with mother, father, and partner. Partial least square modeling suggests that the partner carries forward (mediates) the increased risk of committing sexual abuse associated with insecure/disorganized attachment with the father. A significant partner-mother interaction also suggests that the deleterious effects of attachment to the mother in terms of committing CSA are countered by more secure/organized attachment to the partner. Findings highlight the pivotal role of the romantic partner in IT of CSA. Beyond early intervention, therapy in adulthood aimed at fostering adaptive ways of finding emotional security in the relationship with an adult could thus be encouraged.


Systematic review and case report: Intracranial complications of pediatric sinusitis.
Intracranial complications of rhinosinusitis are rare in the post-antibiotic era. However, due to potentially devastating outcomes, prompt recognition and management are essential. This study aims to perform the first systematic review of the intracranial complications of rhinosinusitis in order to better characterize their clinical presentation, diagnosis, and treatment, and report a case of frontal lobe empyema secondary to pediatric frontoethmoid sinusitis. Ovid MEDLINE, Cochrane Library, and Google Scholar. Full-text, peer-reviewed journal publications from 1947 to January 1, 2015 in English; focus on intracranial complications of sinusitis; pediatric patients (<18 years of age); studies including data on diagnostic workup and treatment. Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. Sixteen studies involving 180 patients were included. An overwhelming majority of patients were young adolescent males (70%). The most common intracranial complications were subdural empyema (49%), epidural abscess (36%), cerebral abscess (21%), and meningitis (10%). Patients most often presented with nonspecific symptoms such as headache, fever, nausea and vomiting. Computed tomography with contrast or magnetic resonance imaging confirmed the diagnosis when intracranial complications were suspected. Typical treatment included surgical incision and drainage, often involving joint neurosurgical and otolaryngological procedures, combined with a long course of antibiotics. The morbidity rate was 27%, and the mortality rate was 3.3%. All studies were retrospective chart reviews, case series or case reports. A review of the currently available literature shows that with a high degree of suspicion, multidisciplinary cooperation and aggressive treatment, favorable outcomes are attainable. The most effective surgical treatment for intracranial complications remains unclear and should be investigated further.


Clinical genomics information management software linking cancer genome sequence and clinical decisions.
Using sequencing information to guide clinical decision-making requires coordination of a diverse set of people and activities. In clinical genomics, the process typically includes sample acquisition, template preparation, genome data generation, analysis to identify and confirm variant alleles, interpretation of clinical significance, and reporting to clinicians. We describe a software application developed within a clinical genomics study, to support this entire process. The software application tracks patients, samples, genomic results, decisions and reports across the cohort, monitors progress and sends reminders, and works alongside an electronic data capture system for the trial's clinical and genomic data. It incorporates systems to read, store, analyze and consolidate sequencing results from multiple technologies, and provides a curated knowledge base of tumor mutation frequency (from the COSMIC database) annotated with clinical significance and drug sensitivity to generate reports for clinicians. By supporting the entire process, the application provides deep support for clinical decision making, enabling the generation of relevant guidance in reports for verification by an expert panel prior to forwarding to the treating physician.


Association of 42 SNPs with genetic risk for cervical cancer: an extensive meta-analysis.
A large number of single nucleotide polymorphisms (SNPs) associated with cervical cancer have been identified through candidate gene association studies and genome-wide association studies (GWAs). However, some studies have yielded different results for the same SNP. To obtain a more comprehensive understanding, we performed a meta-analysis on previously published case-control studies involving the SNPs associated with cervical cancer. Electronic searches of PubMed and Embase were conducted for all publications about the association between gene polymorphisms and cervical cancer. One-hundred and sixty-seven association studies were included in our research. For each SNP, three models (the allele, dominant and recessive effect models) were adopted in the meta-analysis. For each model, the effect summary odds ratio (OR) and 95% CI were calculated. Heterogeneity between studies was evaluated by Cochran's Q test. If the p value of Q test was less than 0.01, a random effect model was used; otherwise, a fixed effect model was used. The results of our meta-analysis showed that: (1) There were 8, 2 and 8 SNPs that were significantly associated with cervical cancer (P < 0.01) in the allele, dominant and recessive effect models, respectively. (2) rs1048943 (CYP1A1 A4889G) showed the strongest association with cervical cancer in the allele effect model (1.83[1.57, 2.13]); in addition, rs1048943 (CYP1A1 A4889G) had a very strong association in the dominant and recessive effect model. (3) 15, 11 and 10 SNPs had high heterogeneity (P < 0.01) in the three models, respectively. (4) There was no published bias for most of the SNPs according to Egger's test (P < 0.01) and Funnel plot analysis. For some SNPs, their association with cervical cancer was only tested in a few studies and, therefore, might have been subjected to published bias. More studies on these loci are required. Our meta-analysis provides a comprehensive evaluation of cervical cancer association studies.


Angiotensin II mediates cell survival through upregulation and activation of the serum and glucocorticoid inducible kinase 1.
The serum- and glucocorticoid-inducible kinase 1 (SGK1) is known to regulate a wide variety of cellular processes, including renal sodium retention and cell survival. Angiotensin II (Ang II) is one of the many signaling molecules capable of regulating SGK1 expression, and is also known to impact cell survival. Here, we examined the role of SGK1 in Ang II-mediated cell survival. We hypothesized that Ang II protects cells from apoptosis by upregulating and activating SGK1. To test this, we examined the effects of Ang II stimulation on SGK1 expression and downstream signaling. We also examined the effects of Ang II treatment and siRNA-mediated SGK1 knockdown on apoptosis after serum starvation. We found that after 2h of Ang II treatment, SGK1 mRNA expression was increased approximately 2-fold. This induction was sensitive to reductions in intracellular calcium levels after pretreatment with BAPTA-AM, but insensitive to the L-type calcium channel blocker verapamil. SGK1 induction was also sensitive to the tyrosine kinase inhibitor genistein. Ang II treatment also caused a rapid increase in the level of phosphorylation of SGK1 at Ser422 and Thr256, and Ser422 phosphorylation was rapamycin-sensitive. We found that Ang II treatment was protective against serum starvation-induced apoptosis, and this protective effect was significantly blunted when SGK1 was silenced via siRNA. Lastly, Ang II induced FOXO3A phosphorylation in an SGK1-dependent manner, thereby reducing the pro-apoptotic actions of FOXO3A. Overall, these results indicate that Ang II upregulates and activates SGK1, leading to increased cell survival via multiple, non-redundant mechanisms.


ggcoverage: an R package to visualize and annotate genome coverage for various NGS data.
Visualizing genome coverage is of vital importance to inspect and interpret various next-generation sequencing (NGS) data. Besides genome coverage, genome annotations are also crucial in the visualization. While different NGS data require different annotations, how to visualize genome coverage and add the annotations appropriately and conveniently is challenging. Many tools have been developed to address this issue. However, existing tools are often inflexible, complicated, lack necessary preprocessing steps and annotations, and the figures generated support limited customization. Here, we introduce ggcoverage, an R package to visualize and annotate genome coverage of multi-groups and multi-omics. The input files for ggcoverage can be in BAM, BigWig, BedGraph and TSV formats. For better usability, ggcoverage provides reliable and efficient ways to perform read normalization, consensus peaks generation and track data loading with state-of-the-art tools. ggcoverage provides various available annotations to adapt to different NGS data (e.g. WGS/WES, RNA-seq, ChIP-seq) and all the available annotations can be easily superimposed with ' + '. ggcoverage can generate publication-quality plots and users can customize the plots with ggplot2. In addition, ggcoverage supports the visualization and annotation of protein coverage. ggcoverage provides a flexible, programmable, efficient and user-friendly way to visualize and annotate genome coverage of multi-groups and multi-omics. The ggcoverage package is available at https://github.com/showteeth/ggcoverage under the MIT license, and the vignettes are available at https://showteeth.github.io/ggcoverage/ .


Detection of altered pain facilitatory and inhibitory mechanisms in patients with knee osteoarthritis by using a simple bedside tool kit (QuantiPain).
Altered pain facilitatory and inhibitory mechanisms have been recognized as an important manifestation in patients with chronic pain, and quantitative sensory testing (QST) can act as a proxy for this process. We have recently developed a simple bedside QST tool kit (QuantiPain) for more clinical use. The purpose of this study was to investigate its test-retest reliability and to evaluate its validity compared with the laboratory-based QST protocols in patients with knee osteoarthritis (OA). QuantiPain consists of 3 items: "pressure algometer" (for pressure pain thresholds [PPTs]), "pinprick" (for temporal summation of pain [TSP]), and "conditioning clamp" (for conditioned pain modulation [CPM]). In experiment-A, intrarater and interrater test-retest reliabilities were investigated in 21 young healthy subjects by using interclass correlation coefficient (ICC). In experiment-B, 40 unilateral painful patients with OA and 40 age-matched, healthy control subjects were included to compare the bedside tool kit against the computerized pressure algometry. In experiment-A, excellent to moderate intrarater and interrater reliabilities were achieved in PPT and TSP (ICC: 0.60-0.92) while the agreements of CPM were good to poor (ICC: 0.37-0.80). In experiment-B, localized and widespread decrease of PPT, facilitated TSP, and impaired CPM was found by using the bedside tool kit in patients with OA compared with controls (P < 0.05). The data were significantly correlated with the established laboratory-based tools (R = 0.281-0.848, P < 0.05). QuantiPain demonstrated acceptable test-retest reliability and assessment validity with the sensitivity to separate patients with painful OA from controls, which has a potential to create more practical approach for quantifying altered pain mechanisms in clinical settings.


The value of thioredoxin family proteins and proliferation markers in dysplastic and malignant gallbladders in patients with primary sclerosing cholangitis.
Patients with primary sclerosing cholangitis (PSC) have an increased risk for biliary and gallbladder malignancy and markers of early malignancy in PSC are lacking. The aims were to evaluate biomarkers to look for premalignancy/malignancy. All available gallbladder specimens (n = 53) in patients with PSC at Karolinska University Hospital between 1985 and 2006 were reviewed. Immunohistochemical staining for p53, Ki-67, Cyclin D1 and the thioredoxin family redox proteins; Thioredoxin reductase 1 (TrxR1), isoform-TrxR1-v.2.3.5, Thioredoxin (Trx1) and Glutaredoxin1 (Grx1) was performed on tissues from patients with carcinoma (n = 6), dysplasia (n = 7) and non-cancerous gallbladder epithelium (n = 6). Dysplasia and carcinoma were found in 16/53 (30%) cases. Inflammation and fibrosis of the gallbladder wall were more common in tissue with gallbladder dysplasia/carcinoma than in benign tissue 12/25 (48%) versus 4/28 (12%) (p < 0.01) and in 13/21 (62%) versus 3/32 (9%) (p < 0.0001), respectively. Immunoreactivity for p53, Ki67, Cyclin D1 was detected in significantly more cases of dysplasia/carcinoma of the gallbladder than in non-cancerous epithelium. 2/19 (11%) of the samples were positive in non-cancerous epithelium versus 7/17 (41%) in dysplasia/carcinoma (p < 0.05) for TrxR1-v.2.3.5. Grx1 was down regulated; more specifically 15/19 (79%) positive cases in non-cancerous epithelium versus 7/17 (41%) in dysplasia/carcinoma. PSC patients have a frequency of gallbladder dysplasia/carcinoma of 30% in operative specimens. The overexpression of TrxR1-v2,3,5 and down regulation of Grx1 in dysplastic gallbladder epithelium suggest that these proteins should be further evaluated as possible future immunohistochemical markers in the early diagnosis of biliary malignancy in PSC.


A Study of Use of Autologous Cartilage in Ossicular Reconstruction.
Ossiculoplasty is one of the commonly done and challenging surgery in otology and has been done using various graft materials. Materials such as autologous cartilage (tragal or conchal) or allograft (homograft) of the same tissue and synthetic materials like polyethylene TORP, ceramic and hydroxyapatite PORP or Titanium prosthesis have been used. But the latter synthetic materials are expensive and have high extrusion rates. Autologous cartilage grafts are selected for Ossiculoplasty as they are easy to harvest at the same site of operation, non-toxic, has less extrusion rate, minimum shrinkage and lateralization. The aim of this study is to know the efficacy of cartilage in ossicular reconstruction and to evaluate hearing outcome. It is a prospective study done on patients attending ENT outpatient department at hospitals attached to J.J.M Medical College, Davanagere. 50 patients were included in the study who underwent ossicular reconstruction either with canal wall up or down mastoidectomy with tympanoplasty. Pre-operative mean ABG was 39.3 db whereas 6 months post-operative mean ABG was 31.6 db with mean ABG improvement of 7.7 db which was statistically significant. Hearing improvement is better in cases where stapes suprastructure was present and also in canal wall up mastoidectomy procedures. This study suggests that cartilage graft (tragal and conchal) is effective in cases of ossicular reconstruction in patients of chronic otitis media and gives good hearing gain. It is also a economical option in developing countries.


A Simple Semi-hydroponic System for Studying Iron Homeostasis in Maize.
Hydroponic-based systems for plant growth allow control of the nutrients that plants take up through the provided nutrient solution. Different formulations of nutrient solutions enable the study of nutrient deficiencies in plants. Here we describe a procedure for setting up a simple semi-hydroponic system to grow maize seedlings. The system can be set up on a small or large scale, depending on the number of individuals studied. A modified nutrient solution is used for growing maize seedlings in iron-replete and iron-depleted conditions. This setup allows for studies of iron-deficiency responses in maize.


Neurosonographic imaging of small-for-gestational-age neonates exposed and not exposed to cocaine and cytomegalovirus.
We sought to prospectively identify the role of neurosonography in the evaluation of a consecutive group of small-for-gestational-age (SGA) neonates, and also to identify the association of neurosonographic findings with cocaine exposure and cytomegalovirus (CMV) infection. Neurosonographic imaging was performed in 180 SGA neonates within 72 hours of birth. Urine samples were screened for CMV and cocaine metabolites (CM) in all cases. Sixty-five neonates (37.5%) had an abnormal neurosonographic appearance. Nine neonates were positive for CMV and 31 neonates were positive for CM. Focal echolucencies (27), ventricular dilation (27), and subependymal hemorrhages (12) were the most common neurosonographic abnormalities. The first two were more common with CM (p < .05). An abnormal neurosonographic pattern was seen more often in SGA neonates with CM (54.8%, 17 of 31; p < .05) and CMV (67%, 6 of 9; p < .01) as compared with the rest (32.6%, 44 of 135; p < .01). Among those without CM or CMV, prematurity was associated with an increased risk for abnormality (p < .001 between groups), specifically subependymal hemorrhage, ventricular dilation, and porencephalic cysts. Five CMV-positive neonates showed periventricular, echogenic foci mainly in the area of the frontal horn. Two new findings with SGA were caudate nucleus echogenicity and a "moth-eaten" appearance of the thalamus, each found in three infants. Neurosonographic imaging is useful in the evaluation of SGA neonates. Focal echolucencies and caudate echogenicity suggest maternal cocaine use, and periventricular echogenic foci strongly suggest fetal CMV infection.


Prediction of secondary structure by evolutionary comparison: application to the alpha subunit of tryptophan synthase.
The amino acid sequences of the a subunits of tryptophan synthase from ten different microorganisms were aligned by standard procedures. The alpha helices, beta strands and turns of each sequence were predicted separately by two standard prediction algorithms and averaged at homologous sequence positions. Additional evidence for conserved secondary structure was derived from profiles of average hydropathy and chain flexibility values, leading to a joint prediction. There is good agreement between (1) predicted beta strands, maximal hydropathy and minimal flexibility, and (2) predicted loops, great chain flexibility, and protein segments that accept insertions of various lengths in individual sequences. The a subunit is predicted to have eight repeated beta-loop-alpha-loop motifs with an extra N-terminal alpha helix and an intercalated segment of highly conserved residues. This pattern suggests that the territory structure of the a subunit is an eightfold alpha/beta barrel. The distribution of conserved amino acid residues and published data on limited proteolysis, chemical modification, and mutagenesis are consistent with the alpha/beta barrel structure. Both the active site of the a subunit and the combining site for the beta 2 subunit are at the end of the barrel formed by the carboxyl-termini of the beta strands.


GABAergic and cholinergic neurons exhibit high-affinity nerve growth factor binding in rat basal forebrain.
We have used dissociated, rat basal forebrain cultures to identify specific cell types that are potentially responsive to nerve growth factor (NGF). Expression of high-affinity NGF binding sites was examined. A subpopulation of cells containing choline acetyltransferase (CAT), the acetylcholine-synthesizing enzyme, exhibited high-affinity binding, employing combined immunocytochemistry and 125I-NGF radioautography. Unexpectedly, a gamma-aminobutyric acid (GABA)-containing cell group also expressed high-affinity binding. These cells that exhibit high-affinity binding appear to be neurons since they stain positively with the neuron marker, neuron-specific enolase, and negatively with the nonneuron marker, glial fibrillary acidic protein. Our observations suggest that NGF may regulate multiple brain systems and functions that have yet to be explored. Conversely, only subsets of cholinergic or GABA neurons expressed high-affinity binding, suggesting that these transmitter populations are composed of differentially responsive subpopulations.


Is assuming additivity of single-metal toxicity thresholds a conservative approach to assessing risk of ecotoxicity from elevated soil concentrations of cobalt, copper, and nickel at contaminated sites?
Multiple metal-impacted soils are often realistic scenarios for risk assessments, but tools to address these are currently lacking. The objective of this work was to evaluate whether assuming concentration addition (CA) of metal mixture effects was conservative for prospective risk assessment of soils that were elevated mainly in Ni and Cu and somewhat with Co, Pb, or As. Observed whole mixture toxicity for field soils with aged metal mixtures was compared to the expected whole mixture toxicity, assuming additivity of prospective single-metal thresholds ("toxic units") for the mixture components. Bioavailability-adjusted single-metal toxicity thresholds expected for those field soils were the median hazard concentration affecting 5% of species (HC5-50) from the predicted no-effect concentration (PNEC) calculator and calculated from the species-specific dose-response multiple linear relationships (MLRs), all from the European Union Registration, Evaluation, Authorisation and Restriction of Chemicals (EU REACH) dossiers for metals. Generic single-metal toxicity thresholds were based on Canadian Council of Ministers of the Environment soil quality guidelines (CCME SQGs) for agricultural soils. Observed toxicity thresholds were from the community-based risk assessments conducted for Port Colborne and Sudbury, Ontario, Canada. Mostly, prospective single-metal toxicity thresholds were protective relative to the observed toxicity, although that was species or ecological process dependent. The bioavailability-adjusted single-metal thresholds were less conservative than the CCME SQG method, even though the former is based on site-specific EC10 values, and the latter is based on generic EC25 values. When within-site variability in soil properties was used to calculate the 5th and 95th CI for the HC5 sum of toxic units (∑TUs), CA was conservative for far fewer endpoints. In addition, the prospective ∑TUs were more conservative predictions of the observed whole mixture toxicities for Port Colborne soils than for Sudbury soils. The most appropriate balance of accuracy and conservatism for identifying low-level risk of the whole mixtures in these soils appeared to be the bioavailability-adjusted HC5-50, which was applicable to many endpoints and 2 quite different exposure concentration ratios. Integr Environ Assess Manag 2021;17:753-766. © 2020 SETAC.


Comparative efficacy of preprandial or postprandial Humalog Mix75/25 versus glyburide in patients 60 to 80 years of age with type 2 diabetes mellitus.
Humalog Mix75/25 (Mix75/25) is a novel premixed insulin containing 75% neutral protamine lispro (an intermediate-acting insulin) and 25% insulin lispro. The purpose of this study was to compare glycemic control and hypoglycemia rates with Mix75/25 versus glyburide, and with preprandial versus postprandial Mix75/25, in patients aged 60 to 80 years with type 2 diabetes mellitus and persistent hyperglycemia on sulfonylurea therapy. In this open-label, 16-week, parallel-group study, patients were randomized to 1 of 2 treatments: glyburide 15 mg/d (or up to the maximum daily dose) or Mix75/25. The Mix75/25 group was randomly subdivided into preprandial (immediately before breakfast and dinner) and postprandial (within 15 minutes after the start of breakfast and dinner) injection subgroups. The primary outcomes were glycemic control and rate of hypoglycemia. A total of 143 patients were randomized; 127 completed the study. The change in glycosylated hemoglobin (HbA(1c)) from baseline to end point was significantly greater with Mix75/25 than with glyburide (mean +/- SEM, -1.14% +/- 0.18% vs -0.36% +/- 0.15%, P = 0.001). HbA(1c) changes with preprandial and postprandial Mix75/25 were not significantly different (-1.20% +/- 0.26% vs -1.08% +/- 0.26%, P = 0.748). Fasting blood glucose (BG), 2-hour postprandial BG, and mean daily BG reductions were greater with Mix75/25 than with glyburide (P < 0.001); preprandial and postprandial Mix75/25 administration did not differ significantly with respect to any of these BG variables. The hypoglycemia rate increased with Mix75/25 by 0.17 +/- 0.02 episodes per patient per 30 days, but there was no change with glyburide (P = 0.077). Body weight increased by 1.02 +/- 0.35 kg with Mix75/25 and decreased by 0.85 +/- 0.18 kg with glyburide (P < 0.001). Compared with glyburide, Mix75/25 significantly improved glycemic control in older patients with type 2 diabetes mellitus, could be administered after meals without compromising glycemic control, and was well tolerated.


Prevalence and characteristics of the aortic ejection sound in adults.
A survey of 1,950 phonocardiograms recorded over a 6-year period revealed 170 (9%) with a distinct aortic ejection sound. All patients were men with a mean age of 61 years (range 29 to 88). Associated clinical features were: aortic stenosis in 28%, history of systemic hypertension in 10%, history of rheumatic fever in 4% and none of these features in 58% of patients. In 141 (83%) of 170 patients the aortic ejection sound occurred simultaneously with or 0.01 second before or after the onset of the rise of the externally recorded carotid pulse. In 37 (66%) of 56 patients who had simultaneous echocardiograms and phonocardiograms recorded, the aortic ejection sound occurred at 0.01 second before or after the maximal opening point of the aortic valve leaflets. Two-dimensional echocardiography was performed in all patients and a bicuspid aortic valve was identified in 38 patients (22%). In 83 patients (49%) 3 cusps were clearly seen. In 49 patients (29%) an accurate determination was not possible. Anatomic examination of 120 consecutive aortic valves at autopsy was performed to identify possible causes of the aortic ejection sound. In 18 (15%) of autopsies fusion of 2 aortic cusps extending greater than or equal to 5 mm from the attachment to the aorta was observed. This abnormality, aortic commissural fusion, may be congenital or acquired. It is concluded that aortic ejection sounds may occur in patients without bicuspid aortic valves and in a variety of clinical conditions. A moderate degree of cuspal fusion may be the cause of the sound.


An antiarrhythmic effect of adenosine during myocardial ischaemia and reperfusion.
Adenosine (10 micrograms kg-1 min-1, infused into the lumen of the left ventricle) and dipyridamole (0.25 mg kg-1 intravenously, a dose that potentiated markedly the fall in arterial pressure in response to bolus doses of adenosine) each reduced the number of extrasystoles which occurred during the first 30 minutes following coronary artery occlusion in anaesthetised greyhound dogs (from 786 +/- 115 in control dogs to 156 +/- 44 in those treated with adenosine and to 388 +/- 167 with dipyridamole). Intracoronary adenosine (1 microgram kg-1 min-1, infused into the ischaemic area) however appeared to increase the number of extrasystoles to 1230 +/- 214. Left ventricular infusion of adenosine reduced the incidence of ventricular fibrillation (from 88 to 43%) when the ischaemic myocardium was perfused at the end of a 40 min occlusion period. In the dose used in this study (10 micrograms kg-1 min-1) adenosine caused a sustained fall in blood pressure but did not alter blood gases. In control dogs the levels of purine derivatives in blood draining the myocardium rendered ischaemic by coronary artery occlusion (local coronary venous samples) increased gradually during the ischaemic period (from 9 +/- 3 microM pre-occlusion to 25 +/- 7 microM post-occlusion). Dipyridamole increased the resting plasma concentration of purines prior to occlusion by approximately 90%; these remained raised for the occlusion period. These results support the suggestion that adenosine may act as a locally produced endogenous antiarrhythmic agent.


Sequential flowering of neighboring goldenrods and the movements of the flower predator Epicauta pennsylvanica.
As neighboring plants flower sequentially, do flower feeders preferentially remain in the area, rather than move to another area with flowering plants? I examined the movements of the meloid beetle Epicauta pennsylvanica, a flower predator specializing on Solidago, in four types of replicated experimental plots - monocultures of Solidago altissima, or S. altissima interplanted with members of the same genus, same family, or different taxonomic orders. I released marked beetles only in the "genus" plots, which contained four species of Solidago, two that bloom before S. altissima. The number of beetles in the genus plots declined steadily as S. altissima came into flower in all the plots; the total number of beetles in all the plots remained fairly constant. I found no evidence that plant neighborhoods affected beetle distribution. Beetles foraging on the early blooming Solidago species did not remain in the genus plots as S. altissima came into flower. In addition, beetles that left the genus plots did not differentially accumulate in any of the other plot types, even though one type of plot was a monoculture with four times the density of S. altissima than the other plots.


Rfv-1 and Rfv-2, two H-2-associated genes that influence recovery from Friend leukemia virus-induced splenomegaly.
Two genes, Rfv-1 and Rfv-2, that influence recovery from Friend virus leukemia have been identified within or adjacent to the mouse major histocompatibility gene complex (H-2). Rfv-1 was determined to be in the H-2G or H-2D regions by testing of mice with recombinations both to the left and right of these regions. Rfv-2 was located either in the H-2K or H-2I regions or in the Tla region. The Rfv-2 effect was not seen in the high recovery H-2D(Rfv-1)b/b mice. This could be an indication that this gene is weaker than Rfv-1 in its influence on leukemia. The influence of H-2 on recovery from leukemia appeared to operate in a gene-dose fashion (H-2b/b greater than H-2a/b greater than H-2a/a).


Prospective, randomized, double-blind trial of curriculum-based training for intracorporeal suturing and knot tying.
Advanced surgical skills such as laparoscopic suturing are difficult to learn in an operating room environment. The use of simulation within a defined skills-training curriculum is attractive for instructor, trainee, and patient. This study examined the impact of a curriculum-based approach to laparoscopic suturing and knot tying. Senior surgery residents in a university-based general surgery residency program were prospectively enrolled and randomized to receive either a simulation-based laparoscopic suturing curriculum (TR group, n=11) or standard clinical training (NR group, n=11). During a laparoscopic Nissen fundoplication, placement of two consecutive intracorporeally knotted sutures was video recorded for analysis. Operative performance was assessed by two reviewers blinded to subject training status using a validated, error-based system to an interrater agreement of >or=80%. Performance measures assessed were time, errors, and needle manipulations, and comparisons between groups were made using an unpaired t-test. Compared with NR subjects, TR subjects performed significantly faster (total time, 526+/-189 seconds versus 790+/-171 seconds; p < 0.004), made significantly fewer errors (total errors, 25.6+/-9.3 versus 37.1+/-10.2; p < 0.01), and had 35% fewer excess needle manipulations (18.5+/-10.5 versus 27.3+/-8.6; p < 0.05). Subjects who receive simulation-based training demonstrate superior intraoperative performance of a highly complex surgical skill. Integration of such skills training should become standard in a surgical residency's skills curriculum.


Tuning Benzylic C-H Functionalization of (Thio)xanthenes with Electrochemistry.
Here, we report a tunable electrochemical benzylic C-H functionalization of (thio)xanthenes with terminal alkynes and nitriles in the absence of any catalyst or external chemical oxidant. The benzylic C-H functionalization can be well controlled by varying the electrochemical conditions, affording the specific coupling products via C-C and C-N bond formation.


Association Between Frailty and Clinical Outcomes and Quality of Life in Older Adults Following Hip Fracture Surgery: A Retrospective Cohort Study.
Hip fracture is a serious event in the older population and is associated with morbidity, mortality, and disability among those who survive. Emerging evidence suggests that frailty is pertinent to the clinical outcomes of older patients with hip fracture. We conducted a retrospective cohort study on older patients (aged ≥65 years) who underwent orthopedic repair with hip fracture under spinal or general anesthesia between June 1, 2019 and May 31, 2020. Demographic, surgical, and anesthetic features; chart-derived frailty index (CFI); occurrence of complications; and length of stay were retrospectively collected by reviewing patients' anesthesia records and medical charts. Patients were divided into 2 groups according to their CFI: high CFI group (CFI, 3-5) and low CFI group (CFI, 0-2). The CFI was calculated as the addition of the patient's conditions, including age >70 years, preoperative body mass index <18.5 kg/m2, hematocrit <35%, albumin <34 g/L, and serum creatinine >176.8 mol/L (2.0 mg/dL). The EuroQol 5-dimensional questionnaire (EQ-5D) was obtained through telephone interviews 12 months after surgery to assess mortality and long-term quality of life. During the study period, 381 patients met the inclusion criteria. Patients in the high CFI group had an increased incidence of delirium by 13.80% (95% confidence interval [CI], 6.31-21.29) (17 [16.6%] vs 8 [2.8%]; P < .001) compared to patients in the low CFI group. More patients in the high CFI group had pneumonia with an increased incidence of 17.71% (95% CI, 7.08-23.34) (40 [39.2%] vs 60 [21.5%]; P < .001) than patients in the low CFI group. Postoperative hospital stay was significantly longer in the high CFI group (8 [6-12] vs 7 [5-10] days; P = .0222) than in the low CFI group. More patients died in the high CFI group 1 year after surgery with an increased mortality of 19.33% (95% CI, 9.47-29.18) (26.4% [23/87] vs 7.1% [16/225]; P < .001) than patients in the low CFI group. A total of 64 patients in the high CFI group and 209 patients in the low CFI group completed the EQ-5D survey 1 year after surgery. The EQ-5D score in the high CFI group was significantly lower than that in the low CFI group (0.63 ± 0.22 vs 0.72 ± 0.22; P = .002). Frailty status is associated with adverse postoperative outcomes, mortality, and low quality of life 12 months after hip fracture surgery in older patients.


Centripetal Versus Centrifugal Facial Nerve Dissection in Superficial Parotidectomy: Comparison of the Two Methods at a Tertiary Care Center.
To compare the intraoperative and postoperative parameters of two techniques of facial nerve dissection: Centripetal vs Centrifugal. Prospective study. Academic tertiary referral centre. In our study, cases of superficial parotidectomy were randomly selected from the outpatient departments of ORL & HNS. Of these patients, 30 underwent standard anterograde dissection group (Group A) and 30 underwent retrograde facial nerve dissection (Group B). All the patients were assessed pre-operatively, intra-operatively and post-operatively to compare the various parameters. The most common complaint in both groups was parotid swelling. The operative time for A was shorter at 2.1 h as compared to 1.9 h min in group B. The resolution of symptoms was nearly comparable in both groups. Facial nerve palsy was slightly more in group B but the difference was not statistically significant. The operative time and post-operative complications were both comparable between AFND and RFND, implying both techniques can be used by surgeons depending on the individual cases.


Aerococcus: an increasingly acknowledged human pathogen.
Aerococci have often been misidentified as streptococci in microbiology laboratories, leading to an underestimation of these bacteria as causes of human infections. An increased awareness of aerococci and the introduction of matrix-assisted laser desorption ionization time-of-flight mass spectrometry, has led to an increased isolation of Aerococcus urinae and Aerococcus sanguinicola from human urine and blood. The two species are found in human urine and can cause urinary tract infections (UTI). Aerococcus urinae can, in older males with underlying urinary tract conditions, cause invasive infections such as urosepsis or infective endocarditis. The prognosis of invasive aerococcal infections appears to be relatively favourable despite the old age of patients and their many comorbidities. Though clinical breakpoints are still not in place, aerococci seem to be sensitive to penicillins, carbapenems and vancomycin. There is synergy between penicillin and aminoglycosides against some A. urinae isolates and this combination is often used in aerococcal infective endocarditis. The treatment of complicated aerococcal UTI is not obvious as many isolates are resistant to fluoroquinolones. In addition, A. urinae is resistant to sulphamethoxazole, and there are methodological problems in the determination of trimethoprim sensitivity. In complicated UTI, ampicillin is probably a safe treatment option, whereas nitrofurantoin is probably effective in uncomplicated UTI. Treatment studies in aerococcal infections are needed as is a better understanding of the natural niches for aerococci and the pathogenesis and clinical course of aerococcal infections.


A sugar-coated strategy to treat a rare neurologic disease provides a blueprint for a decoy glycan therapeutic and a potential vaccine for CoViD-19: An Editorial Highlight for "Selective inhibition of anti-MAG IgM autoantibody binding to myelin by an antigen specific glycopolymer"on page 486.
In a rare neurologic disease known as IgM monoclonal gammopathy the immune system targets a sulfated trisaccharide known as the Human Natural Killer-1 (HNK-1) epitope that comprises a constituent of the myelin sheath known as MAG (myelin-associated glycoprotein). This Editorial highlights a study by Aliu and colleagues in the current issue of the Journal of Neurochemistry, in which the investigators constructed a biodegradable poly-l-lysine backbone with multiple copies of this sulfated HNK-1 trisaccharide. This decoy, poly(phenyl disodium 3-O-sulfo-β-d-glucopyranuronate)-(1→3)-β-d-galactopyranoside, known as PPSGG, removed anti-MAG IgM autoantibodies from the blood, while not activating the immune system. These findings provide a path for the selective removal of a pathogenic set of antibodies that target the myelin sheath resulting in neuropathy. These findings are applicable to a parallel strategy for the generation of polysaccharides similar to those present in the receptor-binding domain of CoViD-19, which might inhibit viral adhesion to its receptor, the angiotensin-converting enzyme-2 (ACE2) protein, thereby impairing cellular uptake of the virus itself. The deployment of complex polysaccharides that mimic actual COVID19 polysaccharides on the spike protein may also provide a feasible structural basis for a vaccine. Carbohydrate mimics, if conjugated to a carrier or backbone, might provoke an immune response to the spike protein. A vaccine that targets critical carbohydrates on COVID19, and then neutralizes the virus would recapitulate a successful strategy employed in other microbial vaccines, like the pneumococcal vaccines and the meningococcal vaccines. These vaccines direct an immune response to complex carbohydrates and successfully prevent life-threatening disease. This paper provides lessons from a rare neurologic disease that may teach us strategies applicable to a global pandemic.


Mapping genomic regions affecting milk traits in Sarda sheep by using the OvineSNP50 Beadchip and principal components to perform combined linkage and linkage disequilibrium analysis.
The detection of regions that affect quantitative traits (QTL), to implement selection assisted by molecular information, remains of particular interest in dairy sheep for which genetic gain is constrained by the high costs of large-scale phenotype and pedigree recording. QTL detection based on the combination of linkage disequilibrium and linkage analysis (LDLA) is the most suitable approach in family-structured populations. The main issue in performing LDLA mapping is the handling of the identity-by-descent (IBD) probability matrix. Here, we propose the use of principal component analysis (PCA) to perform LDLA mapping for milk traits in Sarda dairy sheep. A resource population of 3731 ewes belonging to 161 sire families and genotyped with the OvineSNP50 Beadchip was used to map genomic regions that affect five milk traits. The paternally and maternally inherited gametes of genotyped individuals were reconstructed and IBD probabilities between them were defined both at each SNP position and at the genome level. A QTL detection model fitting fixed effects of principal components that summarize IBD probabilities was tested at each SNP position. Genome-wide (GW) significance thresholds were determined by within-trait permutations. PCA resulted in substantial dimensionality reduction, in fact 137 and 32 (on average) principal components were able to capture 99% of the IBD variation at the locus and genome levels, respectively. Overall, 2563 positions exceeded the 0.05 GW significance threshold for at least one trait, which clustered into 75 QTL regions most of which affected more than one trait. The strongest signal was obtained for protein content on Ovis aries (OAR) chromosome 6 and overlapped with the region that harbours the casein gene cluster. Additional interesting positions were identified on OAR4 for fat content and on OAR11 for the three yield traits. PCA is a good strategy to summarize IBD probabilities. A large number of regions associated to milk traits were identified. The outputs provided by the proposed method are useful for the selection of candidate genes, which need to be further investigated to identify causative mutations or markers in strong LD with them for application in selection programs assisted by molecular information.


A self-powered photoelectrochemical aptamer probe for oxytetracycline based on the use of a NiO nanocrystal/g-C3N4 heterojunction.
A self-powered photoelectrochemical (PEC) aptamer probe is presented for the determination of oxytetracycline (OTC). The assay is based on the use of g-C3N4 and NiO nanocrystals (NCs) which form a heterojunction. The latter was prepared by two-step hydrothermal pyrolysis by using the ionic liquid 1-hydroxyethyl-3-methylimidazole chloride which functions as a morphological template to form NiO NCs. The heterojunction exhibits much better electronic conductivity, wider absorption range, higher electron-hole-separation productivity, and stronger photocurrent compared to plain g-C3N4. The heterojunction was adopted to construct a self-powered PEC aptamer probe for OTC detection. An OTC-binding aptamer was immobilized on the heterojunction and the probe was constructed. The aptamer on the probe binding with OTC can form steric hindrance for transmitting of electrons and cause the PEC signal change depending on the OTC concentration. The photocurrent decreases with increasing OTC concentration in the 0.01 to 100 nM concentration range and its detection limit is 4 pM (at S/N = 3). Graphical abstract Schematic representation of a self-powered photochemical aptamer probe. The probe performs enhanced ability for oxytetracycline (OTC) determination due to the formation of NiO nanocrystals/g-C3N4 (NiO NCs/g-C3N4) heterojunction and the specification recognition of the aptamer.


Multicystic renal cell carcinoma.
This is a case report of a patient with cystic renal cell carcinoma that had all of the ultrasound and computed tomographic features of benign multicystic renal disease. Although this benign entity is a possibility in some patients, thick-wall cystic lesions of the kidney must be considered malignant until proven otherwise and surgery cannot be avoided.


PeptideWitch-A Software Package to Produce High-Stringency Proteomics Data Visualizations from Label-Free Shotgun Proteomics Data.
PeptideWitch is a python-based web module that introduces several key graphical and technical improvements to the Scrappy software platform, which is designed for label-free quantitative shotgun proteomics analysis using normalised spectral abundance factors. The program inputs are low stringency protein identification lists output from peptide-to-spectrum matching search engines for 'control' and 'treated' samples. Through a combination of spectral count summation and inner joins, PeptideWitch processes low stringency data, and outputs high stringency data that are suitable for downstream quantitation. Data quality metrics are generated, and a series of statistical analyses and graphical representations are presented, aimed at defining and presenting the difference between the two sample proteomes.


Modulation of collagen synthesis by fat-storing cells, isolated from CCl4- or vitamin A-treated rats.
In an attempt to elucidate the role of fat-storing cells (FSCs) in liver fibrosis, we investigated the collagen synthesis by FSCs freshly isolated from rats treated with CCl4, with vitamin A, and from untreated rats. FSCs from CCl4-treated rats contained a small number of lipid droplets and an abundant rough endoplasmic reticulum (RER), while those from vitamin A-treated rats showed numerous large lipid droplets and scanty RER. The population doubling times of FSCs isolated from normal, CCl4-treated, and vitamin A-treated rats were 38 +/- 4.3, 24 +/- 2.5, and 48 +/- 6.3 hr, respectively. The rate of collagen synthesis by FSCs from CCl4-treated rats was four- to sixfold enhanced, while collagen synthesis by FSCs from vitamin A-treated rats was suppressed. The ratio of collagen type I to type III produced by FSCs from CCl4 rats was enhanced as compared with control rats (94.7:5.3 vs 87.6:12.4). Therefore, FSCs can be considered to play an important role in the pathogenesis of liver fibrosis.


Step-Down Therapy for Asthma Well Controlled on Inhaled Corticosteroid and Long-Acting Beta-Agonist: A Randomized Clinical Trial.
Stepping down therapy when asthma is well controlled on combination inhaled corticosteroids (ICSs) and long-acting beta-agonists (LABAs) is recommended, but it is not known whether lowering the ICS dose or stopping LABA is superior. To evaluate whether step-down therapy with LABA is superior to one without; and, secondarily, to evaluate whether reducing the ICS dose while maintaining LABA is noninferior to remaining on stable-ICS/LABA. The study was a randomized, double-masked 3-arm parallel group trial in participants aged 12 years or older. Following an 8-week run-in, 459 participants were randomly assigned to continue medium-dose ICS/LABA, reduced-dose ICS/LABA, or ICS alone (LABA-step-off) and followed for 48 weeks. The primary outcome was time to treatment failure, a composite of health care utilization, systemic corticosteroid use, increase in rescue therapy, decline in lung function, or participant or physician decision. Time to treatment failure did not differ significantly between reduced- ICS/LABA and LABA-step-off (hazard ratio, 1.07; 95.3% CI, 0.69-1.65, P = .76). Nor was there a difference between stable-ICS/LABA and reduced-ICS/LABA (hazard ratio, 1.11; 95% CI, 0.70-1.76; P = .67), but the 10% noninferiority margin was exceeded. Lung function declines and hospitalization rates were significantly greater in the LABA-step-off group. The 2 step-down regimens did not differ in terms of treatment failure, although stopping LABA was associated with a decline in lung function and more hospitalizations. There was no evidence to support the noninferiority of reduced-ICS/LABA as compared with stable-ICS/LABA.


Mutagenesis of the cysteine-rich clip domain in the Drosophila patterning protease, Snake.
A common motif found in invertebrate serine proteases involved in immunity and development is the clip domain, proposed to regulate catalytic activity or protein-protein interactions within proteolytic cascades. Snake functions in a cascade that patterns the Drosophila embryo, and provides an accessible model for exploring the structural requirements for clip domain function. We tested Snake zymogens bearing charged-to-alanine mutations in the clip domain for their ability to rescue embryos lacking endogenous Snake and for their interactions by S2 cell co-transfection with upstream Gastrulation Defective and downstream Easter in the protease cascade. Of 13 single and multiple substitutions, one double mutant in a predicted protruding region exhibited a severe defect in embryonic rescue but showed only minimal defects in the co-transfection assay. We discuss implications of these and other results for potential biological roles of the Snake clip domain and for use of the in vitro assay in predicting protease behavior.


Using Theory to Evaluate the Implementation and Integration of an Expanded School Breakfast Program in Rural Midwestern High Schools.
Use Normalization Process Theory to evaluate the implementation and integration of the expanded School Breakfast Program (SBP). Interviews were conducted during the 2014-2015 school year. Normalization Process Theory guided the interview questions. Rural high schools in Minnesota. Interviews were conducted with 12 foodservice directors and 11 principals from the Project Break-Fueling Academics and Strengthening Teens intervention. Four of the 12 schools were in their first year of providing the expanded SBP program to their students whereas 8 were in their second year. Normalization Process Theory was used to evaluate the implementation and integration of the expanded SBP into rural Midwestern high schools. Three members of the research team used NVivo 10 software to code and analyze the interviews. Emerged themes were reported. Modifying the SBP aligned with values of the study participants (coherence). Support was obtained from staff, students, and community members (cognitive participation). Operational work occurred in each school to establish and integrate the modified SBP (collective action). The SBP expansion was assessed through student participation rates (reflexive monitoring). Normalization Process Theory can be used to evaluate the implementation, embedding, and integration of a modified SBP into a school program. This evaluation can support other schools in modifying and embedding their SBPs into their school environment.


Susceptibility of Vibrio parahaemolyticus to disinfectants after prior exposure to sublethal stress.
In the present study, Vibrio parahaemolyticus 690 in phosphate buffered-saline containing 3% NaCl was subjected to sublethal stresses: heat shock at 42 °C for 15 min, acid adaptation at pH 5.0 for 30 min, or cold shock at 20 °C for 4 h. The effect of sublethal stress on the susceptibility of V. parahaemolyticus to a chlorine-containing disinfectant (Clidox-S) and a quaternary ammonium compound (Quatricide) at 25 and 40 °C was investigated. It was found that the sublethal stresses examined enhanced the resistance of V. parahaemolyticus 690 to both disinfectants. Depending on the kinds of sublethal stress, V. parahaemolyticus 690 showed various degrees of enhanced resistance to disinfectants. Furthermore, the phenomenon of enhanced resistance to the disinfectants was more marked at 40 than at 25 °C.


Precise efficiency calibration of an HPGe detector: source measurements and Monte Carlo calculations with sub-percent precision.
With the goal of measuring precise gamma-ray intensities for short-lived (< 5 s) accelerator-produced activities, we have calibrated the efficiency of an HPGe detector between 53 and 1836keV to sub-percent precision with a combination of source measurements and Monte Carlo calculations. Using known or independently measured detector dimensions, we have achieved both relative and absolute agreement (the latter, to 0.1%) between the calculated and measured efficiencies with only two adjustable detector parameters, the thicknesses of the contact dead layers.


Short duration transcranial direct current stimulation (tDCS) modulates verbal memory.
Transcranial direct current stimulation (tDCS) is a noninvasive method of modulating cortical excitability. The aim of this study was to investigate the effects of short-duration tDCS (1.6 seconds per trial) on memory performance, and whether the effects were affected by stimulation administered early or late in a trial. Participants memorize words under anodal and cathodal tDCS to the left dorsolateral prefrontal cortex (DLPFC) in two separate sessions in no-stimulation, early stimulation, and late stimulation trials. Early stimulation occurred during word presentation, whereas late stimulation occurred after word presentation. Early anodal tDCS led to significantly better accuracy and speed in a subsequent recognition test compared to anodal late or no-stimulation conditions. Early cathodal tDCS, on the other hand, led to significantly worse accuracy and speed in a subsequent recognition test compared with cathodal late or no-stimulation conditions. The results of this study suggest that short-duration tDCS can modulate memory performance and highlight the importance of period of stimulation.


Hydrogels Based on Alginates and Carboxymethyl Cellulose with Modulated Drug Release-An Experimental and Theoretical Study.
New hydrogels films crosslinked with epichlorohydrin were prepared based on alginates and carboxymethyl cellulose with properties that recommend them as potential drug delivery systems (e.g., biocompatibility, low toxicity, non-immunogenicity, hemostatic activity and the ability to absorb large amounts of water). The characterization of their structural, morphological, swelling capacity, loading/release and drug efficiency traits proved that these new hydrogels are promising materials for controlled drug delivery systems. Further, a new theoretical model, in the framework of Scale Relativity Theory, was built with to offer insights on the release process at the microscopic level and to simplify the analysis of the release process.


Prolactin and spermatogenesis: new lights on the interplay between prolactin and sperm parameters.
To clarify the relationship between one the most gender-specific hormone, i.e. prolactin (PRL), and semen parameters in men. A retrospective, observational, cohort, real-world study was carried out, enrolling all men performing a semen analysis and PRL examination from 2010 to 2022. For each patient, the first semen analys was extracted, associated to PRL, total testosterone (TT), follicle stimulating hormone (FSH) and luteinizing hormone (LH). Hyperprolactinaemia (>35 ng/mL) was excluded. 1211 subjects were included. PRL serum levels were lower in normozoospermia compared to azoospermia (p = 0.002) and altered semen parameters (p = 0.048) groups. TT serum levels were not different among groups (p = 0.122). Excluding azoospermic men, PRL serum levels were lower in normozoospermic patients, when compared to other groups of semen alterations. An inverse correlation was detected between PRL and sperm concentration. Considering normozospermic subjects, PRL was directly related to both non-progressive sperm motility (p = 0.014) and normal sperm morphology (p = 0.040). Subdiving the cohort in quartiles according to PRL distribution, the highest motilities were observed in the second PRL quartile (8.30-11.10 ng/mL) and asthenozoospermia was significantly predicted by FSH (p < 0.001) and second PRL quartile (p = 0.045). The PRL-spermatogenesis connection seems to be mild, although low-normal PRL levels are associated with the best spermatogenetic profile. PRL serum levels could mirror the immunoregulatory status within the testis, suggesting that there is a sort of 'PRL optimal window' reflecting an efficent spermatogenesis. Alternatively, men with good semen parameters might have a higher central dopaminergic tone resulting in low PRL levels.


[Clinical physiopathology of axoplasmic flow].
Anatomical, biological and patho-physiological characteristics of axoplasmic transport are described in normal conditions and experimental neuropathies.


Comparative studies of the ingestive behaviors produced by microinjections of muscimol into the midbrain raphe nuclei of the ventral tegmental area of the rat.
Microinjection of the GABA-A agonist muscimol into the median (MR) or dorsal (DR) raphe nuclei or the ventral tegmental area (VTA) of non-deprived rats induced intense feeding and drinking in a dose-dependent and site-specific manner. Lower doses of muscimol were required to increase food intake, spillage and water intake with injections into the MR than with injections into the other two sites. These data demonstrate that the MR is a more sensitive site for the elicitation of ingestive behavior than either the DR or the VTA.


Surgical strategies and outcomes of congenital supravalvular aortic stenosis.
Various surgical techniques have been introduced to treat supravalvular aortic stenosis (SVAS). However, there is no consensus upon the optimal approach. This study reviewed our institutional experience in the management of SVAS. Ninety patients undergoing surgery for SVAS were identified between 2009 and 2016. Based on surgical techniques, patients were divided into three groups: McGoon repair (n = 63), Doty repair (n = 24), and Brom repair (n = 3). Median follow-up was 38.5 months (range, 4 months-7.5 years). Patient status, cumulative event-free survival rate, and risk factors for adverse clinical outcomes were assessed. The early mortality rate was 3.3%. There was one late death and two reinterventions. No differences were observed among three surgical groups. Event-free survival was 98.4% at 3 years and 96.5% at 5 years. Diffuse-type SVAS and a preoperative gradient greater than 60 mmHg were risk factors for adverse cardiac remodeling within 6 months post-surgery. Residual aortic stenosis was associated with male gender, preoperative aortic valve stenosis, and a preoperative peak gradient greater than 90 mmHg. Eleven patients (out of 30) who underwent concomitant pulmonary artery patching had a residual pulmonary gradient greater than 40 mmHg. Surgical repair of SVAS can be safely achieved using different techniques, with similar midterm mortality and reintervention rates. Higher preoperative gradient is associated with worse clinical results. Issues regarding surgical timing and concomitant pulmonary artery stenosis need to be further addressed.


Sago cycas-based hierarchical-structured porous carbon for adsorption of acetone vapour: preparation, characterization and performance.
The porous structure and oxygen-containing functional groups of carbon materials play important roles in the adsorption of volatile organic compounds (VOCs). In this study, hierarchical-structured porous carbons (HSPCs) with a large specific surface area and abundant oxygen-containing functional groups were prepared from sago cycas without a template or post-processing for acetone (one of the most common VOCs) adsorption. The micropore volume (0.41-1.15 cm3 g-1) and oxygen-containing functional groups (0.3-1.92 mmol g-1) of HSPCs were manipulated by adjusting the activation temperature. Static adsorption data showed that the HSPC activated at 600 °C (HSPC-600) was superior for acetone adsorption, and a maximum adsorption capacity of 3.75 mmol g-1 was achieved at 25 °C and 0.1 kPa. Breakthrough curves and cyclic adsorption-desorption tests demonstrated the dynamic adsorption capacity and regeneration performance of HSPC-600 were excellent as well. The adsorption isotherms were well described by Langmuir and Langmuir-Freundlich models, indicating the adsorption of acetone on HSPCs is a monolayer adsorption process. Due to electrostatic interaction, hydrogen bond and van der Waals forces between acetone molecules and oxygen-containing functional groups, the adsorption capacity of HSPCs for acetone was significantly improved at low relative pressure. This study may provide a peculiar insight into the development of high-performance acetone adsorbent.


Rapid detection of causative pathogen of peritonitis using in-situ hybridization in a patient with continuous ambulatory peritoneal dialysis.
Continuous ambulatory peritoneal dialysis (CAPD) peritonitis is a common problem in patients on peritoneal dialysis, and the bacteriological diagnosis is important for the treatment of this condition. We used Hybrizep (a method for detecting the genes of bacteria ingested in phagocytes), to detect the causative bacterium in a 50-year-old woman receiving peritoneal dialysis who was admitted our hospital with bacterial peritonitis. The test, using peritoneal dialysis fluid, was positive for Streptococcus epidermidis. Vancomycin administration led to a good outcome for this patient. The Hybrizep method (which is available on the Japanese National Health system only for the diagnosis of septicemia) was a useful diagnostic approach for a patient on peritoneal dialysis who had peritonitis.


Production of polyhydroxybutyrate by Bacillus species isolated from municipal activated sludge.
Plastic wastes are considered to be severe environmental contaminants causing waste disposal problems. Widespread use of biodegradable plastics is one of the solutions, but it is limited by high production cost. Biologic wastewater treatment generates large quantities of biomass as activated sludge. Only a few reports focus on the potential of utilizing resident Bacillus species from activated sludge in polyhydroxbutyrate (PHB) production as well as the production of PHB from food wastes. They have attractive properties such as short generation time, absence of endotoxins, and secretion of both amylases and proteinases that can well utilize food wastes for nutrients, which can further reduce the cost of production of polyhydroxyalkanoates (PHAs). Two PHA-producing strains, HF-1 and HF-2, were isolated from activated sludge. HF-1 outperformed HF-2 in terms of growth and PHB production in hydrolyzed soy and malt wastes. The isolated bacteria was characterized by DNA sequence alignment. Cell extracts of HF-1 were also compared to Bacillus megaterium cell extracts on sodium dodecyl sulfate polyacrylamide gel electrophoresis. The biopolymers accumulated were analyzed by gas chromatography, nuclear magnetic resonance, and Fourier transform infrared methods.


Erythropoietin and erythropoietin receptor expression in normal and disturbed pregnancy.
Erythropoietin (Epo) is known to regulate the number of circulating erythrocytes. Epo receptor (Epo-R) expression is limited to few organs including the uterus. We investigated differences in Epo and Epo-R expression in normal and disturbed first trimester human pregnancy. Placental tissue was obtained from normal human pregnancy, abortion and hydatidiform mole during the first trimester of pregnancy. Epo and Epo-R expression was investigated by Immunohistochemistry and real time PCR (TaqMan). The intensity and distribution patterns of Epo and Epo-R expression were evaluated by using a semi-quantitative method (immunoreactive score) as previously described. Epo-R expression was upregulated in the villous trophoblast (VT) of abortive tissue (p=0.002) as compared to normal pregnancy. This was further confirmed on mRNA level. With regard to mole pregnancy, Epo-R expression was also significantly increased in the VT (p<0.001). In extravillous trophoblasts (EVT), only Epo, not Epo-R expression was significantly increased in abortive tissue (p=0.006) as well as in hydatidiform mole (p=0.041) in comparison to normal pregnancy. Identification of EVT as Epo-and Epo-R-positive cells was obtained by double immunofluorescence with CK7 and Epo/ Epo-R double staining. Both mole pregnancy and abortion were accompanied by an upregulation of Epo (p=0.041; p=0.018) and Epo-R expression (p=0.007; p=0.015) in decidual tissue as compared to normal pregnancy. Within our study we were able to demonstrate increased expression of Epo and Epo-R in abortive tissue and hydatidiform mole. Whether upregulation of Epo and Epo-R is secondary to placental hypoxia as part of the abortion process or a risk factor of its own, needs to be further investigated.


Memory CD4+ and CD8+ T lymphocyte proliferation in vaccinated dairy cows with different histories of Staphylococcus aureus mastitis.
Staphylococcus aureus mastitis constitutes a serious threat to dairy cows. The reasons why available vaccines are not fully effective remain poorly understood; thus, in the present study, we investigated CD4+ and CD8+ T lymphocyte proliferation in dairy cows vaccinated with a polyvalent mastitis vaccine that had distinct precedent Staphylococcus aureus mastitis. We studied 17 S. aureus-infected dairy cows (11 vaccinated and six unvaccinated) and eight vaccinated healthy dairy cows with no previous S. aureus mastitis infections. Flow cytometry was used to assess lymphocyte proliferation using an anti-Ki67 antibody, and monoclonal antibodies were used to identify T cell subsets. S. aureus-infected cows exhibited reduced overall lymphocyte proliferation, including CD4+ T lymphocyte proliferation, and memory lymphocyte proliferation in response to S. aureus isolate stimulus. Immunization did not influence the expansion of blood lymphocyte populations. Furthermore, CD8+ T cells, memory CD8+ T lymphocytes, and effector memory CD8+ T lymphocytes displayed reduced proliferation 21 days after the third vaccine dose compared with before vaccination at time zero. The present data demonstrates an overall negative regulation of the T-cell response suggesting its detrimental impact leading to the persistence of S. aureus intramammary infections. Furthermore, the lack of vaccination effect on T-cell mediated immunity (e.g., proliferation) may be related to poor vaccine efficacy.


Oncolytic adenovirus retargeted to Delta-EGFR induces selective antiglioma activity.
The fact that glioblastomas, which are one of the most devastating cancers, frequently express the Delta-EGFR (epithelial growth factor receptor) also called mutant variant III of EGFR (EGFRvIII) suggests that this cancer cell-specific receptor might serve as an ideal target for cancer therapy. To assess its potential as such a target, we constructed an oncolytic adenovirus with Retargeted Infectivity Via EGFR (Delta-24-RIVER) on the backbone of Delta-24. This new oncolytic adenovirus targets, as Delta-24 does, the disrupted Rb pathway in cancer cells; in addition, this adenovirus has also been retargeted through the abrogation of CAR binding (Y477A mutation in adenoviral fiber protein) and insertion of an EGFRvIII-specific binding peptide in the HI loop of the fiber protein. As compared with Delta-24, Delta-24-RIVER induced EGFRvIII-selective cytotoxicity in U-87 MG isogenic cell lines and in tetracycline-inducible EGFRVIII expressing U-251 MG cells. Accordingly, by tittering the viral progeny and examining fiber protein expression in the above cells, we showed that the replication of this new construct also correlated with EGFRvIII expression. Consistently, immunohistochemistry staining of the adenoviral capsid protein hexon in the virus-treated tumors revealed that the virus replicated more efficiently in EGFRvIII-expressing U-87 MG.DeltaEGFR xenografts than in the tumors grown from U-87 MG cells. Importantly, treatment with Delta-24-RIVER prolonged the survival of animals with intracranial xenografts derived from U-87 MG.DeltaEGFR cells. Therefore, our results constitute the first proof of the direct targeting of a cancer-specific receptor using an oncolytic adenovirus.


Role of the new target specific oral anticoagulants in the management of anticoagulation for cardioversion and atrial fibrillation ablation.
Atrial fibrillation (AF) is the most prevalent arrhythmia in United States. Patients with AF are at increased risk of thromboembolic events. This risk is even higher with cardioversion and ablation, especially during periprocedural period. Conventional approaches using heparin and warfarin have been successfully studied to mitigate thromboembolic risk. However, these approaches are with certain inherent limitations. The advent of new target specific oral anticoagulants (TSOAs) may overcome some of the impediments of conventional therapies and provide an alternative for anticoagulation for cardioversion and AF ablation. We will review the role of the new TSOAs in the current anticoagulation strategies to reduce the risk of thromboembolic events associated with cardioversion and AF ablation.


EQ-5D-3L Norms for the European Older Population: Country-Specific Norms for 15 European Countires Based on the Survey of Health, Ageing, and Retirement in Europe.
This study presents a country-specific 3-level version of EQ-5D population norms for the European older population. Norm data were obtained from the fourth wave of the Survey of Health, Ageing and Retirement in Europe, and determined, for each EQ-5D dimension, the EQ-visual analog scale (EQ-VAS) and EQ-5D index values by 7 age groups and sex for 15 European countries. The EQ-5D index values were calculated using the European VAS value set for all countries. Data resulting from 50 013 older respondents (mean age 65.9 years, range 50-111 years, 55.6% women) revealed an increasing number of self-reported health problems on EQ-5D dimensions and decreasing EQ-VAS scores with increasing age and for women compared with men. There are notable differences between countries in terms of the age gradient, the proportion of respondents in full health, and sex. Across all age groups, problems with pain & discomfort are the most frequent (36%-73% any problems), whereas problems with self-care are the least frequent (3%-31% any problems). The mean EQ-VAS score is 71.2 and the mean European VAS score is 0.79. Given the growing number of older adults and elderly people in Europe, these population norms provide a valuable source of reference data that can be used to compare older adults or patient subgroups to the average of the general elderly population in a similar age or sex group in 15 European countries. The index value results may be further used to assess the burden of disease across older European populations and to identify the unmet needs of targeted older patient populations.


Genetic approaches for the identification of apoptotic components.
Jun amino-terminal kinase (JNK) mediates a physiological stress signal that leads to cell death. However, the role of the JNK pathway in intrinsic cell death execution mechanisms is largely unknown. In a genetic screen for dominant suppressors of Reaper (Rpr)-induced cell death, we identified Drosophila chromosomal regions that contain genes which are homologous to apoptosis signal-regulating kinase (ASK1) and Drosophila tumor necrosis factor receptor-associated factor 1 (DTRAF1). We present evidence that the killer signal initiates the JNK pathway via proteasome-mediated degradation of Drosophila inhibitor of apoptosis protein 1 (DIAP1) to promote cell death.