Cytologic features of metastatic and recurrent melanoma in patients with primary cutaneous desmoplastic melanoma.
Desmoplastic melanoma (DM) is a rare subtype of melanoma characterized by malignant spindle cells associated with prominent fibrocollagenous stroma. Primary melanomas may be entirely desmoplastic ("pure" DM) or exhibit a desmoplastic component admixed with a nondesmoplastic component ("combined" DM). The cytologic features of only 5 cases of DM have been reported previously. Fine-needle biopsy (FNB) specimens from 20 recurrent or metastatic lesions in patients with cutaneous DM and 20 recurrent or metastatic lesions from patients with primary cutaneous non-DM were examined and compared. FNB specimens of patients with DM were less cellular (P = .009) and less often exhibited intranuclear cytoplasmic invaginations (P = .008) and mitotic figures (P = .006) than specimens from patients with non-DM. "Combined" DMs were more commonly composed of epithelioid cells (P = .017) and less often contained bizarre/giant tumor cells (P = .010) than did "pure" DMs. Recurrent and metastatic DM has a range of cytologic appearances. Awareness of the cytologic features and careful clinicopathologic correlation will assist in accurate FNB diagnosis.


The shoot-feeding ecology of three Tomicus species in Yunnan Province, southwestern China.
Three Tomicus pine shoot beetles, T. yunnanensis (Kirkendall and Faccoli) (Coleoptera: Scolytidae), T. minor (Hartig), and T. brevipilosus (Wood and Bright), have been causing serious damage to Yunnan pine ( Pinus yunnanensis (Franchet) (Pinales: Pinaceae)) stands in Yunnan, southwestern China. However, their ability to coexist in the crowns of the same trees during the shoot-feeding phase has not been elucidated. In our study, we investigated and compared the shoot-feeding ecology of the three species of pine shoot beetle in P. yunnanensis in Anning County, Yunnan Province. Shoot-feeding by T. yunnanensis, T. minor, and T. brevipilosus initiated in April, May, and June, and ended in February, April, and May, respectively. Individual T. yunnanensis and T. minor adults fed in shoots for about seven months, and T. brevipilosus for nine months, before initiating reproduction. All three Tomicus species fed in the current-year shoots close to the apical bud. No specific overwintering behavior was observed prior to reproduction. The entrance hole of T. yunnanensis was furthest away from the apical bud, and T. minor was the closest to the apical bud. Differences in the spatial distribution of these shoot-feeding sites might reduce competition among the three beetle species. The long-lasting and overlapping shoot-feeding by the three Tomicus species may reduce the resistance of P. yunnanensis and facilitate the reproduction of these beetles in the trunks of living trees, and thus help explain the severe damage by Tomicus in P. yunnanensis.


Mutations in the fission yeast silencing factors clr4+ and rik1+ disrupt the localisation of the chromo domain protein Swi6p and impair centromere function.
Transcriptional silencing is known to occur at centromeres, telomeres and the mating type region in the nucleus of fission yeast, Schizosaccharomyces pombe. Mating-type silencing factors have previously been shown also to affect transcriptional repression within centromeres and to some extent at telomeres. Mutations in the clr4+, rik1+ and swi6+ genes dramatically reduce silencing at certain centromeric regions and cause elevated chromosome loss rates. Recently, Swi6p was found to co-localise with the three silent chromosomal regions. Here the involvement of clr4+, rik1+ and swi6+ in centromere function is investigated in further detail. Fluorescence in situ hybridisation (FISH) was used to show that, as in swi6 mutant cells, centromeres lag on late anaphase spindles in clr4 and rik1 mutant cells. This phenotype is consistent with a role for these three gene products in fission yeast centromere function. The Swi6 protein was found to be delocalised from all three silent chromosomal regions, and dispersed within the nucleus, in both clr4 and rik1 mutant cells. The phenotypic similarity observed in all three mutants is consistent with the products of both the clr4+ and rik1+ genes being required to recruit Swi6p to the centromere and other silent regions. Mutations in clr4, rik1 and swi6 also result in elevated sensitivity to reagents which destabilise microtubules and show a synergistic interaction with a mutation in the beta-tubulin gene (nda3). These observations suggest that clr4+ and rik1+ must play a role in the assembly of Swi6p into a transcriptionally silent, inaccessible chromatin structure at fission yeast centromeres which is required to facilitate interactions with spindle microtubules and to ensure normal chromosome segregation.


Serum levels of soluble CD163 as a specific marker of macrophage/monocyte activity in sarcoidosis patients.
Monocyte-macrophage lineage cells are the main immunocompetent cells in sarcoidosis. The main cellular elements of sarcoidal granulomas are epithelioid cells and multinucleated giant cells (MGC). MGC are also produced in vitro by human blood monocytes following various stimuli. The in vitro formation of MGC is a useful tool for understanding granulomas. CD163, a scavenger receptor for the hemoglobin-haptoglobin complex, is expressed on monocytes/macrophages and shed into blood in a soluble form (sCD163) after stimulation from Toll-like receptors and oxidative stress. sCD163 serum levels have been reported to increase in inflammatory or infectious conditions. The aim of the present study was to examine the relationship between serum levels of sCD163 and the conventional disease markers of sarcoidosis, and also to evaluate sCD163 levels in culture supernatants following the formation of MGC by human peripheral monocytes in vitro. Twenty sarcoidosis patients and twenty healthy subjects were enrolled in the study. sCD163 serum levels were evaluated using sCD163 ELISA. MGC were formed from peripheral blood monocytes by treatment with supernatant of concanavalin A-stimulated peripheral blood mononuclear cells, and sCD163 levels in the culture supernatants were measured by ELISA. sCD163 serum levels were significantly higher in sarcoidosis patients than in healthy controls and correlated with ACE and soluble interleukin-2 receptor serum levels. sCD163 levels in culture supernatants increased with the production of MGC. sCD163 may be used as a favorable specific marker of macrophage/monocyte activity in order to more clearly understand the disease activity of sarcoidosis.


Restoring the Co magnetic moments at interfacial Co-porphyrin arrays by site-selective uptake of iron.
Magnetochemistry recently emerged as a promising approach to control addressable spin arrays on surfaces. Here we report on the binding, spatial ordering, and magnetic properties of Fe on a highly regular Co-tetraphenylporphyrin (Co-TPP) template and highlight how the Fe controls the magnetism of the Co centers. As evidenced by scanning tunneling microscopy (STM) single Fe atoms attach to the saddle-shape conformers site-selectively in a unique coordination environment offered through a heptamer defined by the Co-N-C-C-C-N cyclic subunit. While the magnetic moment of Co is quenched for bare Co-TPP/Ag(111), the Fe presence revives it. Our X-ray magnetic circular dichroism (XMCD) experiments, complemented by density functional theory (DFT) calculations, evidence a ferromagnetic coupling between the Fe and the Co center concomitant with a complex charge redistribution involving the porphyrin ligand. Thus, we demonstrate an unusual metalloporphyrin coordination geometry that opens pathways to spatially order and engineer magnetic moments in surface-based nanostructures.


The comprehensive complication index: a novel and more sensitive endpoint for assessing outcome and reducing sample size in randomized controlled trials.
To test whether the newly developed comprehensive complication index (CCI) is more sensitive than traditional endpoints for detecting between-group differences in randomized controlled trials (RCTs). A major challenge in RCTs is the choice of optimal endpoints to detect treatment effects. Mortality is no longer a sufficient marker in studies, and morbidity is often poorly defined. The CCI, integrating all complications including their severity in a linear scale ranging from 0 (no complication) to 100 (death), is a new tool, which may be more sensitive than other traditional endpoints to detect treatment effects on postoperative morbidity. The CCI was tested in 3 published RCTs from European centers evaluating pancreas, esophageal and colon resections. To compare the sensitivity of the CCI with traditional morbidity endpoints, for example, presence of any (yes/no) or only the most severe complications, all postoperative events were assessed, and the CCI calculated. Treatment effects and sample size calculations were compared using the CCI and traditional endpoints. Although RCTs failed to show between-group differences using any or most severe complications, the CCI revealed significant differences between treatment groups in 2 RCTs-after pancreas (P=0.009) and esophageal surgery (P=0.014). The CCI in the RCT on colon resections confirmed the absence of between-group differences (P=0.39). The required sample sizes in trials are up to 9 times lower for the CCI than for traditional morbidity endpoints. This study demonstrates superiority of the CCI to traditional endpoints. The CCI may serve as an appealing endpoint for future RCTs and may reduce the sample size.


Calibration of spatial light modulators suffering from spatially varying phase response.
We present a method for converting the desired phase values of a hologram to the correct pixel addressing values of a spatial light modulator (SLM), taking into account detailed spatial variations in the phase response of the SLM. In addition to thickness variations in the liquid crystal layer of the SLM, we also show that these variations in phase response can be caused by a non-uniform electric drive scheme in the SLM or by local heating caused by the incident laser beam. We demonstrate that the use of a global look-up table (LUT), even in combination with a spatially varying scale factor, generally does not yield sufficiently accurate conversion for applications requiring highly controllable output fields, such as holographic optical trapping (HOT). We therefore propose a method where the pixel addressing values are given by a three-dimensional polynomial, with two of the variables being the (x, y)-positions of the pixels, and the third their desired phase values. The coefficients of the polynomial are determined by measuring the phase response in 8 × 8 sub-sections of the SLM surface; the degree of the polynomial is optimized so that the polynomial expression nearly replicates the measurement in the measurement points, while still showing a good interpolation behavior in between. The polynomial evaluation increases the total computation time for hologram generation by only a few percent. Compared to conventional phase conversion methods, for an SLM with varying phase response, we found that the proposed method increases the control of the trap intensities in HOT, and efficiently prevents the appearance of strong unwanted 0th order diffraction that commonly occurs in SLM systems.


Restoration of noncarious tooth defects by dentists in The Dental Practice-Based Research Network.
The authors conducted a study to quantify the reasons for restoring noncarious tooth defects (NCTDs) by dentists in The Dental Practice-Based Research Network (DPBRN) and to assess the tooth, patient and dentist characteristics associated with those reasons. Data were collected by 178 DPBRN dentists regarding the placement of 1,301 consecutive restorations owing to NCTDs. Information gathered included the main clinical reason, other than dental caries, for restoration of previously unrestored permanent tooth surfaces; characteristics of patients who received treatment; dentists' and dental practices' characteristics; teeth and surfaces restored; and restorative materials used. Dentists most often placed restorations to treat lesions caused by abrasion, abfraction or erosion (AAE) (46 percent) and tooth fracture (31 percent). Patients 41 years or older received restorations mainly because of AAE (P < .001). Premolars and anterior teeth were restored mostly owing to AAE; molars were restored mostly owing to tooth fracture (P < .001). Dentists used directly placed resin-based composite (RBC) largely to restore AAE lesions and fractured teeth (P < .001). Among DPBRN practices, AAE and tooth fracture were the main reasons for restoring noncarious tooth surfaces. Pre-molars and anterior teeth of patients 41 years and older are most likely to receive restorations owing to AAE; molars are most likely to receive restorations owing to tooth fracture. Dentists restored both types of NCTDs most often with RBC.


Recombinant human MFG-E8 ameliorates colon damage in DSS- and TNBS-induced colitis in mice.
Inflammatory bowel disease (IBD) is characterized by chronic inflammation of the digestive system and typically requires lifelong medical care. Recombinant human MFG-E8 (rhMFG-E8) is a 364-amino acid protein, which promotes apoptotic cell clearance and reduces inflammation. This study investigates the therapeutic effect of rhMFG-E8 on two well-established mouse models of IBD. Acute mucosal injury leading to colitis was caused by exposing C57BL/6 mice to 4% dextran sodium sulfate (DSS) in the drinking water over 7 days, and BALB/c mice to a single intrarectal dose of 2.75 mg of 2,4,6-trinitrobenzene sulfonic acid (TNBS). Upon clinical onset of colitis (day 2 in the DSS model and day 1 in the TNBS model), mice were treated with daily subcutaneous injections of rhMFG-E8 (60 or 120 μg/kg/day) or vehicle (saline) for 6 days. Treatment with rhMFG-E8 significantly attenuated colitis in both models in a dose-dependent way. Treatment of DSS-induced colitis with rhMFG-E8 (120 μg/kg/day) decreased weight loss by 59%, the colitis severity score by 71%, and colon shrinkage by 49% when compared with vehicle. Similarly, treatment of TNBS-induced colitis with rhMFG-E8 (120 μg/kg/day) decreased weight loss by 97%, the colitis severity score by 82%, and colon shrinkage by 62% when compared with vehicle. In both models, the colons of animals receiving rhMFG-E8 showed marked reduction in neutrophil infiltration, cytokine and chemokine expression, and apoptotic cell counts. In conclusion, rhMFG-E8 ameliorates DSS- and TNBS-induced colitis, suggesting that it has the potential to become a novel therapeutic agent for IBD.


Assessment of acute coronary syndromes: focus on novel biomarkers.
Coronary artery disease (CAD) is the leading cause of mortality in Western Societies and several developing countries. Recent evidence suggests that most detrimental clinical manifestations of CAD, such as acute coronary syndromes (ACS), are the outcome of inflammatory processes that lead to plaque formation and rupture and eventually to ischemia and potentially myocardial necrosis. Neither of the traditionally used biomarkers is thought to be the gold standard in detection of myocardial ischemia or necrosis. A biomarker that could detect quite early the ischemic myocardium as well as define the risk of a future event with high sensitivity and specificity is still lacking. Several biomarkers, implicated in the pathogenesis and clinical evolution of atherosclerosis, have emerged as potent biomarkers for early detection of myocardial ischemia. In the current review, we summarize recent evidence of the most promising biomarkers and discuss their potential role in clinical practice in patients suffering from ACSs.


Patient-Centered Medical Home Exposure and Its Impact on PA Career Intentions.
The transformation of primary care (PC) training sites into patient-centered medical homes (PCMH) has implications for the education of health professionals. This study investigates the extent to which physician assistant (PA) students report learning about the PCMH model and how clinical exposure to PCMH might impact their interest in a primary care career. An electronic survey was distributed to second-year PA students who had recently completed their PC rotation from 12 PA programs. Descriptive statistics and ordered logistic regression analyses were used to characterize the results. A total of 202 second-year PA students completed the survey. When asked about their knowledge of the new health care delivery models, 30% of the students responded they had received instruction about the PCMH. Twenty- five percent of respondents stated they were oriented to new payment structures proposed in the Affordable Care Act and quality improvement principles. Based on their experiences in the primary care clerkship, 64% stated they were likely to pursue a career in primary care, 13% were not likely, and 23% were unsure. Predictors of interest in a primary care career included: (1) age greater than 35 years, (2) being a recipient of a NHSC scholarship, (3) clerkship site setting in an urban cluster of 2,500 to 50,000 people, (4) number of PCMH elements offered at site, and (4) positive impression of team-based care. PA students lack adequate instruction related to the new health care delivery models. Students whose clerkship sites offered greater number of PCMH elements were more interested in pursuing a career in primary care.


"Brain MR spectroscopy in autism spectrum disorder-the GABA excitatory/inhibitory imbalance theory revisited".
Magnetic resonance spectroscopy (MRS) from voxels placed in the left anterior cingulate cortex (ACC) was measured from 14 boys with Autism Spectrum Disorder (ASD) and 24 gender and age-matched typically developing (TD) control group. Our main aims were to compare the concentration of γ-aminobutyric acid (GABA) between the two groups, and to investigate the relationship between brain metabolites and autism symptom severity in the ASD group. We did find a significant negative correlation in the ASD group between Autism Spectrum Screening Questionnaire (ASSQ) and GABA+/Cr, which may imply that severity of symptoms in ASD is associated with differences in the level of GABA in the brain, supporting the excitatory/inhibitory (E/I) imbalance theory. However we did not find a significant difference between the two groups in GABA levels.


Lung cancer screening with low-dose helical CT in Korea: experiences at the Samsung Medical Center.
To determine overall detection rates of lung cancer by low-dose CT (LDCT) screening and to compare histopathologic and imaging differences of detected cancers between high- and low-risk groups, this study included 6,406 asymptomatic Korean adults with >or=45 yr of age who underwent LDCT for lung cancer screening. All were classified into high- (>or=20 pack-year smoking; 3,353) and low-risk (3,053; <20 pack-yr smoking and non-smokers) groups. We compared CT findings of detected cancers and detection rates between high- and low-risk. At initial CT, 35% (2,255 of 6,406) had at least one or more non-calcified nodule. Lung cancer detection rates were 0.36% (23 of 6,406). Twenty-one non-small cell lung cancers appeared as solid (n=14) or ground-glass opacity (GGO) (n=7) nodules. Cancer likelihood was higher in GGO nodules than in solid nodules (p<0.01). Fifteen of 23 cancers occurred in high-risk group and 8 in low-risk group (p=0.215). Therefore, LDCT screening help detect early stage of lung cancer in asymptomatic Korean population with detection rate of 0.36% on a population basis and may be useful for discovering early lung cancer in low-risk group as well as in high-risk group.


Natural soil microbiome variation affects spring foliar phenology with consequences for plant productivity and climate-driven range shifts.
Identifying the potential for natural soil microbial communities to predictably affect complex plant traits is an important frontier in climate change research. Plant phenology varies with environmental and genetic factors, but few studies have examined whether the soil microbiome interacts with plant population differentiation to affect phenology and ecosystem function. We compared soil microbial variation in a widespread tree species (Populus angustifolia) with different soil inoculum treatments in a common garden environment to test how the soil microbiome affects spring foliar phenology and subsequent biomass growth. We hypothesized and show that soil bacterial and fungal communities vary with tree conditioning from different populations and elevations, that this soil community variation influences patterns of foliar phenology and plant growth across populations and elevation gradients, and that transferring lower elevation plant genotypes to higher elevation soil communities delayed foliar phenology, thereby shortening the growing season and reducing annual biomass production. Our findings show the importance of plant-soil interactions that help shape the timing of tree foliar phenology and productivity. These geographic patterns in plant population × microbiome interactions also broaden our understanding of how soil communities impact plant phenotypic variation across key climate change gradients, with consequences for ecosystem functioning.


Genetic population structure of cercariae from an urban foci of Schistosoma mansoni, Brazil.
Rapid urbanization in Brazil has meant that many persons from rural areas where Schistosoma mansoni is endemic have migrated to cities. Discovery of a focus of active transmission in the city of Salvador prompted a citywide survey for active and potential transmission sites. Cercariae shed from infected snails collected from four locations were used to determine how these samples were related and if they were representative of the parasite population infecting humans. Each cercarial collection was greatly differentiated from the others, and diversity was significantly lower when compared with eggs from natural human infections in one site. Egg samples collected 7 years apart in one neighborhood showed little differentiation (Jost's D = 0.01-0.03). Given the clonal nature of parasite reproduction in the snail host and the short-term acquisition of parasites, cercariae from collections at one time point are unlikely to be representative of the diversity in the human population.


Pattern of acute pancreatitis in a tertiary care center in oman.
To describe the epidemiology of acute pancreatitis in a tertiary care center in Oman. A retrospective study evaluated all adults (>18 yrs) who presented with first attack of acute pancreatitis to SQUH from 1(st) of January 2006 to 31(st) of December 2012. One hundred seventy-four patients were found. The mean age of the cohort was 44 ± 11 years. 54% (n=95) were male and 45% (n=79) were female. Majority of the cases were mild (n=131) compared to moderate (n=29) and severe (n=14). Sixty six percent (n=116) of the cases were secondary to alcohol and biliary tract diseases. Alcoholic pancreatitis occurred in 53% of the males whereas biliary pancreatitis occurred in 51% of the females. Pancreatitis recurred in 40% (n=71) of the cases, especially in alcoholic pancreatitis. 56 patients (32%) had complications, especially in the severe cases. The data about acute pancreatitis in Oman was similar to the Western countries. Pancreatitis occurred in males more than females. Alcoholic pancreatitis was more common in males, whereas biliary pancreatitis was more common in females. Alcoholic pancreatitis re-occurred in nearly half of the cases. Severe cases were associated with more complications and recurrence.


Are Heart Failure Management Recommendations and Guidelines Followed in Laboratory Medicine in Europe and North America? The Cardiac Marker Guideline Uptake in Europe (CARMAGUE) Study.
The aim of this survey was to investigate how well heart failure (HF) guidelines for use of natriuretic peptides (NPs) have been implemented in laboratory practice in Europe and North America. In 2013 and 2014, a web-based questionnaire was distributed via North American and European biochemical societies. Questions covered assay performed, reason for method choice, decision limits for HF, and laboratory accreditation status. There were 442 European Union and 91 North American participating laboratories with response rates of 50% and 64% from major or university hospitals, respectively. NP measurements were offered in 67% of European Union and 58% of North American respondents. N-terminal pro-B-type natriuretic peptide (NT-proBNP) was most widely used in Europe (68%) and B-type natriuretic peptide (BNP) was more commonly used (58%) in North America. The most frequent reason for use of a specific assay was the availability of instruments that measure either NT-proBNP (51%) or BNP (67%). For diagnosis of acute HF, NT-proBNP decision limits were diverse; age-dependent limits based on the 2012 European Society of Cardiology (ESC) recommendations were used in only 17% of European sites and 26% of North American sites. For BNP, the guideline-recommended acute HF decision limit of 100 ng/L was better adhered to in Europe (48%) and North America (57%). Surprisingly, similar decision limits were stated for acute and chronic HF by >50% of respondents. NP measurement for HF diagnosis was available in >50% of responding laboratories. However, guideline recommended cutoff values for both acute and chronic HF were still implemented in <30% of participating medical centers.


Arrhythmia discrimination using a smart phone.
We hypothesize that our smartphone-based arrhythmia discrimination algorithm with data acquisition approach reliably differentiates between normal sinus rhythm (NSR), atrial fibrillation (AF), premature ventricular contractions (PVCs) and premature atrial contraction (PACs) in a diverse group of patients having these common arrhythmias. We combine root mean square of successive RR differences and Shannon entropy with Poincare plot (or turning point ratio method) and pulse rise and fall times to increase the sensitivity of AF discrimination and add new capabilities of PVC and PAC identification. To investigate the capability of the smartphone-based algorithm for arrhythmia discrimination, 99 subjects, including 88 study participants with AF at baseline and in NSR after electrical cardioversion, as well as seven participants with PACs and four with PVCs were recruited. Using a smartphone, we collected 2-min pulsatile time series from each recruited subject. This clinical application results show that the proposed method detects NSR with specificity of 0.9886, and discriminates PVCs and PACs from AF with sensitivities of 0.9684 and 0.9783, respectively.


Potential of tumor-suppressive miR-596 targeting LGALS3BP as a therapeutic agent in oral cancer.
The incidence and mortality statistics for oral squamous cell carcinoma (OSCC) were 10th and 12th, respectively, in human cancers diagnosed worldwide in 2008. In this study, to identify novel tumor-suppressive microRNAs (TS-miRNAs) and their direct targets in OSCC, we performed methylation-based screening for 43 miRNAs encoded by 46 miRNA genes located within 500 bp downstream of 40 CpG islands and genome-wide gene expression profiling in combination with a prediction database analysis, respectively, in 18 cell lines, resulting in the identification of a novel TS-miRNA miR-596 directly targeting LGALS3BP/Mac-2 BP/90K. DNA hypermethylation of CpG island located 5'-upstream of miR-596 gene was frequently observed in OSCC cell lines (100% of 18 cell lines) and primary OSCC cases (46.2 and 76.3% of 26 Japanese and 38 Thais primary cases, respectively) in a tumor-specific manner. The ectopic transfection of double-stranded RNA (dsRNA) mimicking miR-596 or specific small interfering RNA for LGALS3BP significantly induced growth inhibition and apoptosis in cell lines lacking miR-596 expression or overexpressing LGALS3BP, respectively, in a manner associated with a suppression of ERK1/2 phosphorylation. Moreover, we also mention the effect of dsRNA mimicking miR-596 on the growth of an OSCC cell line in vivo. Our findings define a central role for miR-596 in OSCC and suggest the potential of miR-596 as an anticancer agent for miRNA replacement therapy in OSCC.


Gliding Associated Proteins Play Essential Roles during the Formation of the Inner Membrane Complex of Toxoplasma gondii.
The inner membrane complex (IMC) of apicomplexan parasites is a specialised structure localised beneath the parasite's plasma membrane, and is important for parasite stability and intracellular replication. Furthermore, it serves as an anchor for the myosin A motor complex, termed the glideosome. While the role of this protein complex in parasite motility and host cell invasion has been well described, additional roles during the asexual life cycle are unknown. Here, we demonstrate that core elements of the glideosome, the gliding associated proteins GAP40 and GAP50 as well as members of the GAPM family, have critical roles in the biogenesis of the IMC during intracellular replication. Deletion or disruption of these genes resulted in the rapid collapse of developing parasites after initiation of the cell cycle and led to redistribution of other glideosome components.


Safety evaluation of the food enzyme phospholipase A1 from the genetically modified Aspergillus niger strain NZYM-FP.
The food enzyme phospholipase A1 (phosphatidycholine 1-acylhydrolase EC 188.8.131.52) is produced with the genetically modified Aspergillus niger strain NZYM-FP by Novozymes A/S. The genetic modifications do not give rise to safety concerns. The food enzyme is free from viable cells of the production organism and its DNA. The phospholipase A1 food enzyme is intended to be used for degumming of fats and oils. Since residual amounts of Total Organic Solids (TOS) are removed by the purification steps applied during degumming, dietary exposure estimation was not considered necessary. Genotoxicity tests did not raise safety concerns. The repeated dose 90-day oral toxicity was assessed by means of a repeated dose 90-day oral toxicity study in rats. The Panel identified a no-observed-adverse-effect level (NOAEL) of 1,356 mg TOS/kg body weight (bw) per day, the highest dose tested. Similarity of the amino acid sequence to those of known allergens was searched and no match was found. The Panel considered that, under the intended conditions of use, the risk of allergic sensitisation and elicitation reactions upon dietary exposure cannot be excluded, but the likelihood for this to occur is considered to be low. Based on the data provided and the removal of TOS during the degumming of fats and oils, the Panel concluded that this food enzyme does not give rise to safety concerns under the intended conditions of use.


Risk of premenopausal breast cancer in association with occupational exposure to polycyclic aromatic hydrocarbons and benzene.
This study examined the relationship between risk of premenopausal breast cancer and occupational exposure to benzene and polycyclic aromatic hydrocarbons (PAH) and whether the proposed relationship between PAH and breast cancer differed by tumor estrogen receptor (ER) status. In a case-referent study of premenopausal breast cancer, occupational histories and other information were obtained through interviews, and job-exposure matrices were used to assess exposure to PAH and benzene. A dose-response relationship for the probability of exposure to benzene [low: odds ratio (OR) 1.64, 95% confidence interval (95% CI) 0.64-4.21; high: OR 1.95, 95% CI 1.14-3.33) and to PAH (low: OR 1.56, 95% CI 0.78-3.12; high: OR 2.40, 95% CI 0.96-6.01). Risk increased with duration of exposure to benzene, but not to PAH. A dose-response relationship was not evident for the intensity of exposure to benzene or to PAH. When analyses were stratified by tumor ER status, PAH exposure was related to a greater increase in the risk of ER-positive (OR 2.27, 95% CI 1.14-4.54) than ER-negative (OR 1.12, 95% CI 0.47-2.64) breast cancer. Risk of ER-positive, but not ER-negative, tumors increased with the probability of exposure to PAH. The findings suggest an association between risk and occupational exposure to benzene. Although it was difficult to study PAH independently of benzene, there was some suggestion of an association between PAH exposure and ER-positive tumors. These data should be interpreted with caution because of the limitations of this study, including low-response rates and small numbers of exposed persons.


Increased serum tumor necrosis factor receptor-associated factor-6 expression in patients with non-metastatic triple-negative breast cancer.
Obesity appears to be associated with an increased risk of breast cancer (BC) and an inferior oncological outcome at the time of diagnosis, with poor outcomes most prominent in cases of triple-negative BC (TNBC). The present study analyzed serum tumor necrosis factor (TNF)-like weak inducer of apoptosis (TWEAK) and TNF receptor associated factor-6 (TRAF) protein expression levels in 48 patients with non-metastatic BC and 26 obese control patients (without BC). The mean age of the cohort was 52.5 years (range, 35-78 years) and the patients had a median body mass index of 33.5 kg/m2 (range, 30-47 kg/m2). In the study population, 27.1% of BC patients were triple negative and 70.8% were hormone receptor (HR)-positive. Median serum TRAF6 expression was 0.90 ng/ml (range, 0.55-1.53 ng/ml) in the 13 TNBC patients and 0.63 ng/ml (range, 0.49-1.22 ng/ml) in the 35 HR-positive BC patients; thus, TRAF6 expression was significantly higher in the TNBC patients compared with the obese control group (0.90 vs. 0.73 ng/ml; P=0.033). Furthermore, median serum TRAF6 expression levels were significantly higher in HR-negative patients compared with HR-positive patients (0.83 vs. 0.62 ng/ml; P=0.002). The present study demonstrated that serum TRAF6 expression levels were increased in TNBC and HR-negative patients with non-metastatic BC compared with HR- and human epidermal growth factor receptor 2-positive cases or the obese healthy control group. Therefore, elevated TRAF6 expression may be a poor prognostic factor in non-metastatic BC. In addition, we propose that progesterone (PR) negativity may be a more useful poor prognosis factor than estrogen receptor (ER) negativity, as TRAF6 expression levels were higher in the PR-negative patients compared with the ER-negative patients.


Dynamics of brush border remodeling induced by enteropathogenic E. coli.
Enteropathogenic Escherichia coli (EPEC) induces dramatic remodeling of enterocyte brush borders, a process that includes microvillar effacement and actin pedestal formation. Although the Arp2/3 complex is involved in formation of a branched actin network within pedestals, the fate of parallel actin bundles in microvilli during infection remains unclear. Here, we find that in polarized intestinal epithelial cells, EPEC stimulates long-range microvillar dynamics, pulling protrusions toward sites of bacterial attachment in a process mediated by the adhesion molecule protocadherin-24. Additionally, retraction of the EPEC bundle forming pilus stimulates directed elongation of nearby microvilli. These processes lead to coalescence of microvilli and incorporation of the underlying parallel actin bundles into pedestals. Furthermore, stabilization of microvillar actin bundles delays pedestal formation. Together, these results suggest a model where EPEC takes advantage of pre-existing actin filaments in microvillar core bundles to facilitate pedestal formation.


Renal biopsy in lupus patients with low levels of proteinuria.
Early and accurate detection of kidney involvement in systemic lupus erythematosus (SLE) improves outcomes. Renal biopsy is required for definitive diagnosis of lupus nephritis (LN). In the absence of acute renal failure (ARF), moderate levels of proteinuria (> 1000 mg/24 h) have been recommended by some to justify biopsy. We investigated whether patients with lower levels of proteinuria without ARF have significant renal disease and should be routinely biopsied. We retrospectively evaluated 21 SLE patients with 24-h urine protein < 1000 mg who underwent kidney biopsies. Indications for biopsy included new-onset proteinuria, increasing proteinuria, or hematuria (> 5 red blood cells per high power field). No patient had ARF. Sixteen of 21 (77%) biopsies were diagnostic of LN: 3 class II, 10 class III (5 superimposed class V), 2 class IV (one superimposed class V), and one with class V. One patient had thrombotic microangiopathy. The remaining 4 (23%) patients had non-lupus renal disease. Thirteen patients with class III or greater LN required alterations in therapeutic regimen because of biopsy findings. Of 7 patients without hematuria at the time of biopsy, 4 (57%) had class III, IV, or V LN. One patient without hematuria and < 500 mg/24 h proteinuria had class III LN. We found significant renal involvement (Class III, IV, or V LN) in SLE patients with < 1000 mg proteinuria with or without hematuria. Our findings suggest that biopsy be strongly considered in this patient population.


What is happening during case deliberations in clinical ethics committees? A pilot study.
Clinical ethics consultation services have been established in many countries during recent decades. An important task is to discuss concrete clinical cases. However, empirical research observing what is happening during such deliberations is scarce. To explore clinical ethics committees' deliberations and to identify areas for improvement. A pilot study including observations of committees deliberating a paper case, semistructured group interviews, and qualitative analysis of the data. Nine hospital ethics committees in Norway. Key elements of the deliberations included identifying the ethical problems; exploring moral values and principles; clarifying key concepts and relevant legal regulation; exploring medical facts, the patient's situation, the therapists' perspective, analogous clinical situations, professional uncertainties, the patient's and relatives' perspective, and clinical communication; identifying the involved parties and how to involve them; identifying possible courses of action, and possible conclusion and follow-up. The various elements were closely interwoven. The content and conclusions varied and seemed to be contingent on the committee members' interpretations, experience and knowledge. Important aspects of a clinical ethics deliberation were sometimes neglected. When the committees used a deliberation procedure and a blackboard, the deliberations tended to become more systematic and transparent. Many of the committees were insecure about how to include the involved parties and how to document the deliberations. Clinical ethics committees may provide an important arena for multidisciplinary discussions of complex clinical ethics challenges. However, this seems to require adequate composition, adoption of transparent deliberation procedures, and targeted training.


Dendritic epidermal T cells regulate skin antimicrobial barrier function.
The epidermis, the outer layer of the skin, forms a physical and antimicrobial shield to protect the body from environmental threats. Skin injury severely compromises the epidermal barrier and requires immediate repair. Dendritic epidermal T cells (DETC) reside in the murine epidermis where they sense skin injury and serve as regulators and orchestrators of immune responses. Here, we determined that TCR stimulation and skin injury induces IL-17A production by a subset of DETC. This subset of IL-17A-producing DETC was distinct from IFN-γ producers, despite similar surface marker profiles. Functionally, blocking IL-17A or genetic deletion of IL-17A resulted in delayed wound closure in animals. Skin organ cultures from Tcrd-/-, which lack DETC, and Il17a-/- mice both exhibited wound-healing defects. Wound healing was fully restored by the addition of WT DETC, but only partially restored by IL-17A-deficient DETC, demonstrating the importance of IL-17A to wound healing. Following skin injury, DETC-derived IL-17A induced expression of multiple host-defense molecules in epidermal keratinocytes to promote healing. Together, these data provide a mechanistic link between IL-17A production by DETC, host-defense, and wound-healing responses in the skin. These findings establish a critical and unique role of IL-17A-producing DETC in epidermal barrier function and wound healing.


Understanding human functioning using graphical models.
Functioning and disability are universal human experiences. However, our current understanding of functioning from a comprehensive perspective is limited. The development of the International Classification of Functioning, Disability and Health (ICF) on the one hand and recent developments in graphical modeling on the other hand might be combined and open the door to a more comprehensive understanding of human functioning. The objective of our paper therefore is to explore how graphical models can be used in the study of ICF data for a range of applications. We show the applicability of graphical models on ICF data for different tasks: Visualization of the dependence structure of the data set, dimension reduction and comparison of subpopulations. Moreover, we further developed and applied recent findings in causal inference using graphical models to estimate bounds on intervention effects in an observational study with many variables and without knowing the underlying causal structure. In each field, graphical models could be applied giving results of high face-validity. In particular, graphical models could be used for visualization of functioning in patients with spinal cord injury. The resulting graph consisted of several connected components which can be used for dimension reduction. Moreover, we found that the differences in the dependence structures between subpopulations were relevant and could be systematically analyzed using graphical models. Finally, when estimating bounds on causal effects of ICF categories on general health perceptions among patients with chronic health conditions, we found that the five ICF categories that showed the strongest effect were plausible. Graphical Models are a flexible tool and lend themselves for a wide range of applications. In particular, studies involving ICF data seem to be suited for analysis using graphical models.


Effect of antihypertensive therapy on focal stroke in spontaneously hypertensive rats.
Spontaneously hypertensive rats subjected to focal cerebral ischemia develop larger infarcts than normotensive rat strains. To determine whether antihypertensive therapy decreases infarct volume in hypertensive rats, 60 13-week-old animals were treated with 20 mg/kg hydralazine added daily to the drinking water for 1.5, 6, 10, or 16 weeks and then subjected to focal cerebral ischemia by tandem right common carotid artery and middle cerebral artery occlusion. Blood pressure in the treated groups was substantially lower than that in untreated groups after 1 week of hydralazine therapy and remained lower for the entire treatment period in all four experiments. Mean infarct volume in spontaneously hypertensive rats treated for 10 (p = 0.02) or 16 (p = 0.005) weeks, but not 1.5 or 6 weeks, was significantly less than that in the untreated controls. The percentage reduction of infarct volume in animals treated for 10 and 16 weeks was similar. This study demonstrates that antihypertensive therapy decreases infarct volume in hypertensive rats subjected to focal cerebral ischemia. This treatment effect appears to be dependent on the duration of therapy, and the magnitude of the treatment effect seems to plateau by 10 weeks of therapy.


The Dnmt3L ADD Domain Controls Cytosine Methylation Establishment during Spermatogenesis.
A critical aspect of mammalian gametogenesis is the reprogramming of genomic DNA methylation. The catalytically inactive adaptor Dnmt3L is essential to ensuring this occurs correctly, but the mechanism by which it functions is unclear. Using gene targeting to engineer a single-amino-acid mutation, we show that the Dnmt3L histone H3 binding domain (ADD) is necessary for spermatogenesis. Genome-wide single-base-resolution DNA methylome analysis of mutant germ cells revealed overall reductions in CG methylation at repetitive sequences and non-promoter CpG islands. Strikingly, we also observe an even more severe loss of non-CG methylation, suggesting an unexpected role for the ADD in this process. These epigenetic deficiencies were coupled with defects in spermatogonia, with mutant cells displaying marked changes in gene expression and reactivation of retrotransposons. Our results demonstrate that the Dnmt3L ADD is necessary for Dnmt3L function and full reproductive fitness.


The microvascular system of the striate and extrastriate visual cortex of the macaque.
In functional neuroimaging, neurovascular coupling is used to generate maps of hemodynamic changes that are assumed to be surrogates of regional neural activation. The aim of this study was to characterize the microvascular system of the primate cortex as a basis for understanding the constraints imposed on a region's hemodynamic response by the vascular architecture, density, as well as area- and layer-specific variations. In the macaque visual cortex, an array of anatomical techniques has been applied, including corrosion casts, immunohistochemistry, and cytochrome oxidase (COX) staining. Detailed measurements of regional vascular length density, volume fraction, and surface density revealed a similar vascularization in different visual areas. Whereas the lower cortical layers showed a positive correlation between the vascular and cell density, this relationship was very weak in the upper layers. Synapse density values taken from the literature also displayed a very moderate correlation with the vascular density. However, the vascular density was strongly correlated with the steady-state metabolic demand as measured by COX activity. This observation suggests that although the number of neurons and synapses determines an upper bound on an area's integrative capacity, its vascularization reflects the neural activity of those subpopulations that represent a "default" mode of brain steady state.


Double-strand break repair in the absence of RAD51 in yeast: a possible role for break-induced DNA replication.
In wild-type diploid cells of Saccharomyces cerevisiae, an HO endonuclease-induced double-strand break (DSB) at the MAT locus can be efficiently repaired by gene conversion using the homologous chromosome sequences. Repair of the broken chromosome was nearly eliminated in rad52delta diploids; 99% lost the broken chromosome. However, in rad51delta diploids, the broken chromosomes were repaired approximately 35% of the time. None of these repair events were simple gene conversions or gene conversions with an associated crossover, instead, they created diploids homozygous for the MAT locus and all markers in the 100-kb region distal to the site of the DSB. In rad51delta diploids, the broken chromosome can apparently be inherited for several generations, as many of these repair events are found as sectored colonies, with one part being repaired and the other part being lost the broken chromosome. Similar events occur in about 2% of wild-type cells. We propose that a broken chromosome end can invade a homologous template in the absence of RAD51 and initiate DNA replication that may extend to the telomere, 100 or more kb away. Such break-induced replication appears to be similar to recombination-initiated replication in bacteria.


Imaging of Myocardial Fibrosis in Patients with End-Stage Renal Disease: Current Limitations and Future Possibilities.
Cardiovascular disease in patients with end-stage renal disease (ESRD) is driven by a different set of processes than in the general population. These processes lead to pathological changes in cardiac structure and function that include the development of left ventricular hypertrophy and left ventricular dilatation and the development of myocardial fibrosis. Reduction in left ventricular hypertrophy has been the established goal of many interventional trials in patients with chronic kidney disease, but a recent systematic review has questioned whether reduction of left ventricular hypertrophy improves cardiovascular mortality as previously thought. The development of novel imaging biomarkers that link to cardiovascular outcomes and that are specific to the disease processes in ESRD is therefore required. Postmortem studies of patients with ESRD on hemodialysis have shown that the extent of myocardial fibrosis is strongly linked to cardiovascular death and accurate imaging of myocardial fibrosis would be an attractive target as an imaging biomarker. In this article we will discuss the current imaging methods available to measure myocardial fibrosis in patients with ESRD, the reliability of the techniques, specific challenges and important limitations in patients with ESRD, and how to further develop the techniques we have so they are sufficiently robust for use in future clinical trials.


Clinical results of a surgical technique using endobuttons for complete tendon tear of pectoralis major muscle: report of five cases.
We herein describe a surgical technique for the repair of complete tear of the pectoralis major (PM) tendon using endobuttons to strengthen initial fixation. Five male patients (3 judo players, 1 martial arts player, and 1 body builder) were treated within 2 weeks of sustaining complete tear of the PM tendon. Average age at surgery and follow-up period were 28.4 years (range, 23-33) and 28.8 months (range, 24-36). A rectangular bone trough (about 1 × 4 cm) was created on the humerus at the insertion of the distal PM tendon. The tendon stump was introduced into this trough, and fixed to the reverse side of the humeral cortex using endobuttons and non-absorbable suture. Clinical assessment of re-tear was examined by MRI. Shoulder range of motion (ROM), outcome of treatment, and isometric power were measured at final follow-up. There were no clinical re-tears, and MRI findings also showed continuity of the PM tendon in all cases at final follow-up. Average ROM did not differ significantly between the affected and unaffected shoulders. The clinical outcomes at final follow-up were excellent (4/5 cases) or good (1/5). In addition, postoperative isometric power in horizontal flexion of the affected shoulder showed complete recovery when compared with the unaffected side. Satisfactory outcomes could be obtained when surgery using the endobutton technique was performed within 2 weeks after complete tear of the PM tendon. Therefore, our new technique appears promising as a useful method to treat complete tear of the PM tendon.


Changes in cell wall biomechanical properties in the xyloglucan-deficient xxt1/xxt2 mutant of Arabidopsis.
The main load-bearing network in the primary cell wall of most land plants is commonly depicted as a scaffold of cellulose microfibrils tethered by xyloglucans. However, a xyloglucan-deficient mutant (xylosyltransferase1/xylosyltransferase2 [xxt1/xxt2]) was recently developed that was smaller than the wild type but otherwise nearly normal in its development, casting doubt on xyloglucan's role in wall structure. To assess xyloglucan function in the Arabidopsis (Arabidopsis thaliana) wall, we compared the behavior of petiole cell walls from xxt1/xxt2 and wild-type plants using creep, stress relaxation, and stress/strain assays, in combination with reagents that cut or solubilize specific components of the wall matrix. Stress/strain assays showed xxt1/xxt2 walls to be more extensible than wild-type walls (supporting a reinforcing role for xyloglucan) but less extensible in creep and stress relaxation processes mediated by α-expansin. Fusicoccin-induced "acid growth" was likewise reduced in xxt1/xxt2 petioles. The results show that xyloglucan is important for wall loosening by α-expansin, and the smaller size of the xxt1/xxt2 mutant may stem from the reduced effectiveness of α-expansins in the absence of xyloglucan. Loosening agents that act on xylans and pectins elicited greater extension in creep assays of xxt1/xxt2 cell walls compared with wild-type walls, consistent with a larger mechanical role for these matrix polymers in the absence of xyloglucan. Our results illustrate the need for multiple biomechanical assays to evaluate wall properties and indicate that the common depiction of a cellulose-xyloglucan network as the major load-bearing structure is in need of revision.


Effect of sorafenib on sperm count and sperm motility in male Swiss albino mice.
The issue of male germ line mutagenesis and the effects on developmental defects in the next generation has become increasingly high profile over recent years. Mutagenic substance affects germinal cells in the testis. Since the cells are undergoing different phases of cell division and maturation, it is an ideal system to study the effect of chemotherapeutic agents. There are lacunae in the literature on the effect of sorafenib on gonadal function. With background, a study was planned to evaluate the effects of sorafenib on sperm count and sperm motility in male Swiss albino mice. Male Swiss albino mice were used for the study. The animals were segregated into control, positive control (PC) and three treatment groups. PC received oral imatinib (100 mg/kg body weight) and treatment groups received 25, 50, and 100 mg/kg body weight of sorafenib orally for 7 consecutive days at intervals of 24 h between two administrations. The control group remained in the home cage for an equal duration of time to match their corresponding treatment groups. The animals were sacrificed at the end of 1(st), 2(nd), 4(th), 5(th), 7(th), and 10(th) weeks after the last exposure to drug, respectively. Sperm suspensions were prepared and introduced into a counting chamber. Total sperm count and motility were recorded. There was a significant decrease in sperm count and sperm motility by sorafenib which was comparable with the effect of PC imatinib. Sorafenib adversely affects sperm count and sperm motility which are reversible after discontinuation of treatment.


Using a patient-centered approach to benefit-harm assessment in treatment decision-making: a case study in uveitis.
Synthesizing evidence from comparative effectiveness trials can be difficult because multiple outcomes of different importance are to be considered. The goal of this study was to demonstrate an approach to conducting quantitative benefit-harm assessment that considers patient preferences. We conducted a benefit-harm assessment using data from the Multicenter Uveitis Steroid Treatment Trial that compared corticosteroid implant versus systemic corticosteroids and immunosuppression in non-infectious intermediate, posterior, and panuveitis. We focused on clinical outcomes considered important to patients, including visual acuity, development of cataracts/glaucoma, need for eye surgery, prescription-requiring hypertension, hyperlipidemia, and infections. Patient preferences elicited in a recent survey were then incorporated into our assessment of the benefit-harm balance. Benefit-harm metrics were calculated for each time point that summarized the numbers of outcomes, caused or prevented by implant therapy versus systemic therapy if 1000 patients were treated. The benefit-harm metric was -129 (95% confidence interval: -242 to -14), -317 (-436 to -196), -390 (-514 to -264), and -526 (-687 to -368) at 6, 12, 18, and 24 months follow up, respectively, suggesting that systemic therapy may have a better benefit-harm balance. However, measures of quality of life for patients treated with implant therapy were found to be better than patients treated with systemic therapy over the same time period. Results of benefit-harm assessment were different from the prospectively collected quality of life data during trial follow up. Future studies should explore the reasons for such discrepancies and the strength and weakness of each method to assess treatment benefits and harms.


The method used to culture host cells (Sf9 cells) can affect the qualities of baculovirus budding particles expressing recombinant proteins.
Budded virus (BV) particles of baculovirus (Autographa californica nucleopolyhedrovirus, AcNPV) are harvested from the supernatant of liquid culture of Sf9 host cells by ultracentrifugation. Using polyacrylamide gel electrophoresis, Western blot and transmission electron microscopy (TEM) of BV samples fractionated closely by sucrose density gradient centrifugation, we observed that BVs exhibited different qualities depending on whether they had been harvested from the supernatant from a standing (static), shaking (suspension), or standing/shaking (pre-/post-infection) culture of Sf9 cells. The amount of BV protein apparently increased in the order of standing, standing/shaking, and shaking procedure, and the yield of intact particles showed an opposite trend. TEM observation clearly showed that appropriate fractions of the standing and standing/shaking cultures contained more intact BV particles than those from the shaking culture. These results suggest that the qualities of recombinant BV particles may be related to the culture conditions of the host cells.


Teaching systems-based practice to residents by using independent study projects.
The changing health care marketplace requires new graduates to be familiar with complex health systems. Since 1999 the Oregon Health & Science University internal medicine residency program has offered a seminar-based health systems curriculum, but residents lacked an opportunity to actively apply learned concepts. To achieve this goal, the authors developed a second curricular component, the Health Systems Independent Study Project (HSISP). The HSISP is a self-directed assessment of a health care system or delivery issue residents complete in their third year. The curriculum was implemented in 2001 with goals of gaining a focused understanding of a health systems concept and building related skills. Residents present their study projects to all curriculum participants. Topics addressed so far include adherence to coronary artery disease guidelines in a defined population, screening strategies for lung cancer, resident burnout, and many others. Residents have found that these projects enhance their systems knowledge, help them link systems-based-practice concepts to clinical practice, and foster opportunities for networking and early career development.


Genetic variation of genes involved in dihydrotestosterone metabolism and the risk of prostate cancer.
Dihydrotestosterone (DHT) is an important factor in prostate cancer (PCA) genesis and disease progression. Given PCA's strong genetic component, we evaluated the possibility that variation in genes involved in DHT metabolism influence PCA risk. We investigated copy number variants (CNV) and single nucleotide polymorphisms (SNP). We explored associations between CNV of uridine diphospho-glucuronosyltransferase (UGT) genes from the 2B subclass, given their prostate specificity and/or involvement in steroid metabolism and PCA risk. We also investigated associations between SNPs in genes (HSD3B1, SRD5A1/2, and AKR1C2) involved in the conversion of testosterone to DHT, and in DHT metabolism and PCA risk. The population consisted of 426 men (205 controls and 221 cases) who underwent prostate-specific antigen screening as part of a PCA early detection program in Tyrol, Austria. No association between CNV in UGT2B17 and UGT2B28 and PCA risk was identified. Men carrying the AA genotype at SNP rs6428830 (HSD3B1) had an odds ratio (OR) of 2.0 [95% confidence intervals (95% CI), 1.1-4.1] compared with men with GG, and men with AG or GG versus AA in rs1691053 (SRD5A1) had an OR of 1.8 (95% CI, 1.04-3.13). Individuals carrying both risk alleles had an OR of 3.1 (95% CI, 1.4-6.7) when compared with men carrying neither (P = 0.005). Controls with the AA genotype on rs7594951 (SRD5A2) tended toward higher serum DHT levels (P = 0.03). This is the first study to implicate the 5alpha-reductase isoform 1 (SRD5A1) and PCA risk, supporting the rationale of blocking enzymatic activity of both isoforms of 5alpha-reductase for PCA chemoprevention.


Alcohol dependence and reproductive onset: findings in two Australian twin cohorts.
Although early alcohol use is a strong predictor of future alcohol problems and adolescent drinking is associated with risky sexual behavior predictive of early childbearing, reproductive dysfunctions associated with delayed childbearing have been reported in adult drinkers. We examine the relationship between lifetime history of alcohol dependence (AD) and timing of first childbirth across reproductive development. Data were drawn from two cohorts of Australian twins born between 1893 and 1964 (3634 female and 1880 male twins) and between 1964 and 1971 (3381 female and 2748 male twins). Survival analyses were conducted using Cox proportional hazards regression models predicting age at first childbirth from AD, with sociodemographic characteristics, regular smoking, history of psychopathology, and family and childhood risks included as control variables in adjusted models. Results suggest alcoholic women in both cohorts show overall delayed reproduction, with little effect of AD on timing of first reproduction in men. Effects of AD are particularly strong for women in the older cohort, where AD is associated with 73% decreased likelihood of first childbirth after age 29 [hazard ratio (HR) = 0.27, 95% CI: 0.10-0.75]. In adjusted models, effects reduce only slightly (HR = 0.29, 95% CI: 0.11-0.80). For women in the young cohort, AD is associated with delayed reproduction after age 24, with 40% decreased likelihood of first childbirth (HR = 0.60, 95% CI: 0.48-0.75). AD remains predictive in adjusted models, but without age interaction (HR = 0.72, 95% CI: 0.62-0.85). Findings of delayed reproductive onset in alcoholic women are consistent with alcohol-related reproductive dysfunctions, although underlying mechanisms remain largely unknown. To better understand AD differences in reproductive onset, continued research on both biological and psychosocial risks is needed.


Whole-Body MRI: Comparison of Its Capability for TNM Staging of Malignant Pleural Mesothelioma With That of Coregistered PET/MRI, Integrated FDG PET/CT, and Conventional Imaging.
The purpose of this study was to compare the diagnostic accuracy of whole-body MRI, coregistered FDG PET/MRI, integrated FDG PET/CT, and conventional imaging examination including bone scintigraphy, contrast-enhanced brain MRI, and CT for malignant pleural mesothelioma (MPM) staging according to the new International Association for the Study of Lung Cancer (IASLC) system. The study subjects were 23 consecutively registered patients with MPM (15 men, eight women; mean age, 68 years for both sexes) who had prospectively undergone whole-body FDG PET/CT, whole-body MRI, conventional radiologic examination, surgical or conventional treatments, pathologic examination, and follow-up conventional imaging examinations between January 2011 and December 2017. TNM staging was evaluated by two independent readers. Kappa statistics and chi-square tests were used for evaluation agreements on each factor and clinical stage between each method and final diagnosis. The diagnostic accuracy of each method was statistically compared by use of McNemar test. The kappa values for each factor between each method and final diagnosis were significant (p < 0.0001) and ranged between 0.33 and 0.91. Kappa values between final diagnosis and stage evaluation were also significant (p < 0.0001) and ranged between 0.57 and 0.91. The diagnostic accuracy of N and stage assessment of whole-body MRI and FDG PET/MRI was significantly higher than that of conventional imaging examination (N factor, p < 0.05; stage, p < 0.05). The diagnostic accuracy of whole-body MRI, FDG PET/MRI, and FDG PET/CT for TNM stage assessment based on the new IASLC MPM staging system is greater than that of conventional imaging examination.


The effects of insects, nutrients, and plant invasion on community structure and function above-and belowground.
Soil nutrient availability, invasive plants, and insect presence can directly alter ecosystem structure and function, but less is known about how these factors may interact. In this 6-year study in an old-field ecosystem, we manipulated insect abundance (reduced and control), the propagule pressure of an invasive nitrogen-fixing plant (propagules added and control), and soil nutrient availability (nitrogen added, nitrogen reduced and control) in a fully crossed, completely randomized plot design. We found that nutrient amendment and, occasionally, insect abundance interacted with the propagule pressure of an invasive plant to alter above-and belowground structure and function at our site. Not surprisingly, nutrient amendment had a direct effect on aboveground biomass and soil nutrient mineralization. The introduction of invasive nitrogen-fixing plant propagules interacted with nutrient amendment and insect presence to alter soil bacterial abundance and the activity of the microbial community. While the larger-scale, longer-term bulk measurements such as biomass production and nutrient mineralization responded to the direct effects of our treatments, the shorter-term and dynamic microbial communities tended to respond to interactions among our treatments. Our results indicate that soil nutrients, invasive plants, and insect herbivores determine both above-and belowground responses, but whether such effects are independent versus interdependent varies with scale.


Antithrombotic Therapy After Left Atrial Appendage Occlusion in Patients With Atrial Fibrillation.
Pivotal trials of percutaneous left atrial appendage occlusion (LAAO) used specific postprocedure treatment protocols. This study sought to evaluate patterns of postprocedure care after LAAO with the Watchman device in clinical practice and compare the risk of adverse events for different discharge antithrombotic strategies. We evaluated patients in the LAAO Registry of the National Cardiovascular Data Registry who underwent LAAO with the Watchman device between 2016 and 2018. We assessed adherence to the full postprocedure trial protocol including standardized follow-up, imaging, and antithrombotic agents and then evaluated the most commonly used antithrombotic strategies and compared the rates and risk of adverse events at 45 days and 6 months by means of multivariable COX frailty regression. Among 31,994 patients undergoing successful LAAO, only 12.2% received the full postprocedure treatment protocol studied in pivotal trials; the most common protocol deviations were with discharge antithrombotic medications. The most common discharge medication strategies were warfarin and aspirin (36.9%), direct oral anticoagulant (DOAC) and aspirin (20.8%), warfarin only (13.5%), DOAC only (12.3%), and dual antiplatelet therapy (5.0%). In multivariable Cox frailty regression, the adjusted risk of any adverse event through the 45-day follow-up visit were significantly lower for discharge on warfarin alone (HR: 0.692; 95% CI: 0.569-0.841) and DOAC alone (HR: 0.731; 95% CI: 0.574-0.930) compared with warfarin and aspirin. Warfarin alone retained lower risk at the 6-month follow-up. In contemporary U.S. practice, practitioners rarely used the full U.S. Food and Drug Administration-approved postprocedure treatment protocols studied in pivotal trials of the Watchman device. Discharge after implantation on warfarin or DOAC without concomitant aspirin was associated with lower risk of adverse outcomes.


EGFR Mutation Testing of non-squamous NSCLC: Impact and Uptake during Implementation of Testing Guidelines in a Population-Based Registry Cohort from Northern New Zealand.
Since 2013, clinical practice guidelines recommend EGFR mutation testing of non-squamous NSCLC to select advanced-stage patients for first-line treatment using EGFR-TKIs. We aimed to determine population-based trends in the real-world uptake and impact in routine practice of these recently updated testing guidelines. A population-based observational study was conducted of notifications to the New Zealand Cancer Registry of patients eligible for EGFR testing diagnosed in northern New Zealand between January 2010 and April 2014. The main study variable was EGFR mutation testing. Main outcome measures (overall survival and dispensing of EGFR-TKIs) were extracted from prospectively archived electronic databases until October 2015. The population-based cohort of 1857 patients had an average age of 70 years. Most had adenocarcinoma and metastatic disease at diagnosis. EGFR testing was undertaken in 500 patients (27%) with mutations detected in 109 patients (22%). EGFR testing increased during the period of study from <5% to 67% of patients (P < 0.0001). Full uptake of testing by all eligible patients was limited by a lack of availability of specimens for testing and variable testing referral practices. The proportion of patients treated with EGFR-TKIs decreased during the same time period, both among untested patients (from 12.2% to 2.8% (P < 0.0001)) and in the population as a whole (from 13.7% to 10.6% (P < 0.05)). EGFR testing was associated with prolonged overall survival (Adjusted HR = 0.76 (95% CI, 0.65-0.89) Log-rank P < 0.0001) due at least in part to the much longer overall survival achieved by mutation-positive patients, of whom 79% received EGFR-TKIs. Compared to untested EGFR-TKI-treated patients, mutation-positive EGFR-TKI-treated patients received EGFR-TKIs for longer, and survived longer both from the start of EGFR-TKI treatment and date of their diagnosis. In this real world setting, high uptake of EGFR testing was achieved and associated with major changes in EGFR-TKI prescribing and improved health outcomes. Modifiable factors determined testing uptake. Study registration ACTRN12615000998549.


Health Sensors, Smart Home Devices, and the Internet of Medical Things: An Opportunity for Dramatic Improvement in Care for the Lower Extremity Complications of Diabetes.
The prevalent and long-neglected diabetic foot ulcer (DFU) and its related complications rank among the most debilitating and costly sequelae of diabetes. With the rise of the Internet of medical things (IoMT), along with smart devices, the med-tech industry is on the cusp of a home-care revolution, which could also create opportunity for developing effective solutions with significant potential to reduce DFU-associated costs and saving limbs. This article discusses potential applications of IoMT to the DFU patient population and beyond. To better understand potential opportunities and challenges associated with implementing IoMT for management of DFU, the authors reviewed recent relevant literatures and included their own expert opinions from a multidisciplinary point of view including podiatry, engineering, and data security. The IoMT has opened digital transformation of home-based diabetic foot care, as it enables promoting patient engagement, personalized care and smart management of chronic and noncommunicable diseases through individual data-driven treatment regimens, telecommunication, data mining, and comprehensive feedback tailored to individual requirements. In particular, with recent advances in voice-activated commands technology and its integration as a part of IoMT, new opportunities have emerged to improve the patient's central role and responsibility in enabling an optimized health care ecosystem. The IoMT has opened new opportunities in health care from remote monitoring to smart sensors and medical device integration. While it is at its early stage of development, ultimately we envisage a connected home that, using voice-controlled technology and Bluetooth-radio-connected add-ons, may augment much of what home health does today.


Natural communities of Achromatium oxaliferum comprise genetically, morphologically, and ecologically distinct subpopulations.
The diversity and ecology of natural communities of the uncultivated bacterium Achromatium oxaliferum were studied by use of culture-independent approaches. 16S rRNA gene sequences were PCR amplified from DNA extracted from highly purified preparations of cells that were morphologically identified as A. oxaliferum present in freshwater sediments from three locations in northern England (Rydal Water, Jenny Dam, Hell Kettles). Cloning and sequence analysis of the PCR-amplified 16S rRNA genes revealed that multiple related but divergent sequences were routinely obtained from the A. oxaliferum communities present in all the sediments examined. Whole-cell in situ hybridization with combinations of fluorescence-labelled oligonucleotide probes revealed that the divergent sequences recovered from purified A. oxaliferum cells corresponded to genetically distinct Achromatium subpopulations. Analysis of the cell size distribution of the genetically distinct subpopulations demonstrated that each was also morphologically distinct. Furthermore, there was a high degree of endemism in the Achromatium sequences recovered from different sediments; identical sequences were never recovered from different sampling locations. In addition to ecological differences that were apparent between Achromatium communities from different freshwater sediments, the distribution of different subpopulations of Achromatium in relation to sediment redox profiles indicated that the genetically and morphologically distinct organisms that coexisted in a single sediment were also ecologically distinct and were adapted to different redox conditions. This result suggests that Achromatium populations have undergone adaptive radiation and that the divergent Achromatium species occupy different niches in the sediments which they inhabit.


Post-Transformation IGHV-IGHD-IGHJ Mutations in Chronic Lymphocytic Leukemia B Cells: Implications for Mutational Mechanisms and Impact on Clinical Course.
Analyses of IGHV gene mutations in chronic lymphocytic leukemia (CLL) have had a major impact on the prognostication and treatment of this disease. A hallmark of IGHV-mutation status is that it very rarely changes clonally over time. Nevertheless, targeted and deep DNA sequencing of IGHV-IGHD-IGHJ regions has revealed intraclonal heterogeneity. We used a DNA sequencing approach that achieves considerable depth and minimizes artefacts and amplification bias to identify IGHV-IGHD-IGHJ subclones in patients with prolonged temporal follow-up. Our findings extend previous studies, revealing intraclonal IGHV-IGHD-IGHJ diversification in almost all CLL clones. Also, they indicate that some subclones with additional IGHV-IGHD-IGHJ mutations can become a large fraction of the leukemic burden, reaching numerical criteria for monoclonal B-cell lymphocytosis. Notably, the occurrence and complexity of post-transformation IGHV-IGHD-IGHJ heterogeneity and the expansion of diversified subclones are similar among U-CLL and M-CLL patients. The molecular characteristics of the mutations present in the parental, clinically dominant CLL clone (CDC) differed from those developing post-transformation (post-CDC). Post-CDC mutations exhibit significantly lower fractions of mutations bearing signatures of activation induced deaminase (AID) and of error-prone repair by Polη, and most of the mutations were not ascribable to those enzymes. Additionally, post-CDC mutations displayed a lower percentage of nucleotide transitions compared with transversions that was also not like the action of AID. Finally, the post-CDC mutations led to significantly lower ratios of replacement to silent mutations in VH CDRs and higher ratios in VH FRs, distributions different from mutations found in normal B-cell subsets undergoing an AID-mediated process. Based on these findings, we propose that post-transformation mutations in CLL cells either reflect a dysfunctional standard somatic mutational process or point to the action of another mutational process not previously associated with IG V gene loci. If the former option is the case, post-CDC mutations could lead to a lesser dependence on antigen dependent BCR signaling and potentially a greater influence of off-target, non-IG genomic mutations. Alternatively, the latter activity could add a new stimulatory survival/growth advantage mediated by the BCR through structurally altered FRs, such as that occurring by superantigen binding and stimulation.


Bimodal electric tissue ablation (BETA) compared with the Cool-Tip RFA system.
Bimodal electric tissue ablation (BETA) incorporates the process of electrolysis into radiofrequency ablation (RFA) to increase the size of tissue ablation. This study investigated whether BETA could increase the efficacy of the Cool-Tip RF system (Covidien, Boulder, CO, USA) to produce larger ablations. It also investigated whether applying electrolysis only during the pretreatment phase (called electrochemical treatment (ECT)/RFA group) is as effective as BETA (where electrolysis was used during both the pretreatment and RFA phases). A Cool-Tip RF system (Covidien) was used to test three types of ablations (RFA, BETA, and ECT/RFA) in a pig liver model. In BETA, 9 V of direct current was provided for 10 min, after which the RF generator was started and both electrical circuits were allowed to run concurrently. In ECT/RFA, however, the direct current circuit was switched off after 10 min of pretreatment and only RFA was performed as described above. Ablation sizes were measured in three dimensions. The size of ablations (transverse diameter A and B) produced by BETA and ECT/RFA was significantly larger compared with standard RFA (P < 0/001). BETA also created larger ablations compared with ECT/RFA (P < 0.001). BETA could improve the efficacy of the Cool-Tip RF system (Covidien) to achieve larger ablations. The increased tissue hydration improved delivery of electrical energy to the tissues and delayed the process of desiccation, thus allowing the ablation process to continue for longer periods of time to produce larger ablations. BETA could be used to treat larger liver tumours more effectively than standard RFA.


Ultrasound-guided peripheral nerve blocks in high-risk patients, requiring lower limb (Above and below knee) amputation.
A case series of five high-risk patients with lower limb ischemia, sepsis, altered coagulation, and multi-organ dysfunction requiring emergent or urgent lower limb amputation is presented. Use of peripheral nerve blocks for below and above knee amputations is quite uncommon which provides better hemodynamic stability and pain management, especially in the very sick patients. The surgeries were successfully carried out under ultrasound-guided combined femoral and sciatic nerve blocks. All five patients obtained adequate level of block at the area of amputation. There were no complications related to anesthetic management. Patients remained hemodynamically stable and pain-free during surgery and postoperatively. Combined femoral-sciatic nerve block under ultrasound guidance is safe and satisfactory alternative anesthetic technique for such patients.


Evidence that the caterpillar salivary enzyme glucose oxidase provides herbivore offense in solanaceous plants.
The insect salivary enzyme glucose oxidase (GOX) can inhibit wound-inducible nicotine production in tobacco, Nicotiana tabacum. We examined whether salivary gland extracts of Helicoverpa zea lacking active GOX could still suppress nicotine in tobacco, Nicotiana tabacum, and whether GOX could suppress wound-inducible defenses of another Solanaceous plant, tomato Lycopersicon esculentum. Tobacco leaves were wounded with a cork borer and treated with water, salivary gland extracts with active GOX (SxG), or salivary gland extracts with inactive GOX (SxI). After three days, leaves treated with SxG had significantly less nicotine than all other wounded treatments. Neonates that fed on the terminal leaves of tobacco plants treated with SxG had significantly higher survival than neonates that fed on leaves treated with either SxI or water. This evidence supports the assertion that GOX is the salivary factor responsible for the suppression of tobacco plant nicotine production by H. zea saliva. Results for the NahG tobacco plants, which lack salicylic acid (SA) due to a transgene for bacterial SA hydroxylase, indicate that suppression of nicotine by GOX does not require SA. However, tobacco leaves that were wounded and treated with SxG had significantly higher levels of the SA-mediated PR-1a protein than leaves treated with SxI or water. Leaves of tomato plants wounded with scissors and then treated with SxG had trypsin inhibitor levels that were moderately lower than plants wounded and treated with purified GOX, water, or SxI. However, all the wounded tomato leaves irrespective of treatment resulted in lower caterpillar growth rates than the non-wounded tomato leaves. Glucose oxidase is the first insect salivary enzyme shown to suppress wound-inducible herbivore defenses of plants.


Nursing care for people with delirium superimposed on dementia.
Nursing and healthcare is changing in response to an ageing population. There is a renewed need for holistic nursing to provide clinically competent, appropriate and timely care for patients who may present with inextricably linked mental and physical health requirements. This article explores the dichotomy in healthcare provision for 'physical' and 'mental' health, and the unique role nurses have when caring for people with delirium superimposed on dementia (DSD). Delirium is prevalent in older people and recognised as 'acute brain failure'. As an acute change in cognition, it presents a unique challenge when occurring in a person with dementia and poses a significant risk of mortality. In this article, dementia is contrasted with delirium and subtypes of delirium presentation are discussed. Nurses can recognise DSD through history gathering, implementation of appropriate care and effective communication with families and the multidisciplinary team. A simple mnemonic called PINCH ME (Pain, INfection, Constipation, deHydration, Medication, Environment) can help identify potential underlying causes of DSD and considerations for care planning. The mnemonic can easily be adapted to different clinical settings and a fictitious scenario is presented to show its application in practice.


An evaluation of the effect of a topical product containing C-xyloside and blueberry extract on the appearance of type II diabetic skin.
Diabetes is a multisystem disease caused by the presence of chronic hyperglycemia, which leads to increased oxidative stress. Many of the changes observed in type II diabetic patients can be traced to the increased production of advanced glycation end products, also known as AGEs. AGEs are produced as a result of a nonenzymatic reaction with glucose interacting with proteins, lipids, and nucleic acids. AGEs are also present in normal skin with advancing age and contribute to the senescence of many body organs, including the skin. This research evaluated the effect of a topical product formulation containing blueberry extract, an AGE inhibitor, and C-xyloside, a GAG synthesis stimulator, applied twice daily on the hand, arm, and facial skin of 20 type II diabetic females. Diabetic skin was chosen for evaluation because AGEs are found in increased concentration in diabetic skin, representing a model for accelerated aging. This single-center study enrolled 20 female type II diabetics aged 55+ years with mild to moderate fine lines, wrinkles, and hyperpigmentation on the face and hands. Subjects used the study product on their face, hand, and inner forearm twice daily for 12 weeks. Ordinal grading on a 4-point scale (0 = none, 1 = mild, 2 =moderate, 3 = severe) of facial fine lines, wrinkles, firmness, radiance, skin tone, skin smoothness, hyperpigmentation, creping, density, sagging, and overall appearance was performed by the investigator at baseline, week 4, week 8, and week 12. Tolerability, subject assessments, digital photography, AGE measurements, skin caliper measurements, and corneometry were also performed at each time point. 19/20 subjects successfully completed the study. The presence of AGEs was documented by skin autofluorescence. The 12-week duration of the study was insufficient to measure a change in skin AGEs, but longer application of the study product might produce different results. No tolerability issues were noted. There was a statistically significant increase in skin caliper measurements on the face (P = 0.004) and arm (P = 0.014) as well as corneometry measurements (P < 0.001) consistent with enhanced moisturization at week 12. The dermatologist investigator also found statistically significant improvement in fine lines (P = 0.01), firmness (P = 0.011), radiance (P < 0.001), skin tone (P = 0.014), skin smoothness (P < 0.001), creping (P < 0.004), and overall appearance (P < 0.001). This study examined a topical product containing an AGE inhibitor and a GAG synthesis stimulator designed for the unique needs of diabetic skin.


DICER1 regulates endometrial carcinoma invasion via histone acetylation and methylation.
Endometrial carcinoma (EC) is one of the most common gynecologic malignancy, but molecular mechanisms of the development and progression of EC remain unclear. Here we showed that the expression of DICER1 was negatively associated with the level of histone methylation, histone acetylation and PRC2 components SUZ12 and EZH2 in EC cells. In addition, knockdown of DICER1 significantly downregulated miR-200b and let-7i, which may then regulate their targets SUZ12 and EZH2. Furthermore, knockdown of DICER1 remarkably suppressed the expression of epithelial cell marker E-cadherin, induced the expression of mesenchymal cell marker Vimentin, and promoted the invasion of EC cells. In conclusion, our data suggest that DICER1 suppresses SUZ12 and EZH2 via affecting their upstream miRNA synthesis, and inhibits epithelial-mesenchymal transition(EMT) and invasion of EC cells via histone modification.


Impacts of solid fuel use versus smoking on life expectancy at age 30 years in the rural and urban Chinese population: a prospective cohort study.
The impact of solid fuel use on life expectancy (LE) in less-developed countries remains unclear. We aimed to evaluate the potential impact of household solid fuel use on LE in the rural and urban Chinese population, with the effect of smoking as a reference. We used data from China Kadoorie Biobank (CKB) of 484,915 participants aged 30-79 free of coronary heart disease, stroke, or cancer at baseline. Analyses were performed separately for solid fuel use for cooking, solid fuel use for heating, and smoking, with participants exposed to the other two sources excluded. Solid fuels refer to coal and wood, and clean fuels refer to electricity, gas, and central heating. We used a flexible parametric Royston-Parmar model to estimate hazard ratios of all-cause mortality and predict LE at age 30. Totally, 185,077, 95,228, and 230,995 participants were included in cooking-, heating-, and smoking-related analyses, respectively. During a median follow-up of approximately 12.1 years, 12,725, 7,531, and 18,878 deaths were recorded in the respective analysis. Compared with clean fuel users who reported cooking with ventilation, participants who used solid fuels with ventilation and without ventilation had a difference in LE (95% confidence interval [CI]) at age 30 of -1.72 (-2.88, -0.57) and -2.62 (-4.16, -1.05) years for men and -1.33 (-1.85, -0.81) and -1.35 (-2.02, -0.67) years for women, respectively. The difference in LE (95% CI) for heating was -2.23 (-3.51, -0.95) years for men and -1.28 (-2.08, -0.48) years for women. In rural men, the LE reduction (95% CI) related to solid fuel use for cooking (-2.55; -4.51, -0.58) or heating (-3.26; -6.09, 0.44) was more than that related to smoking (-1.71; -2.54, -0.89). Conversely, in urban men, the LE reduction (95% CI) related to smoking (-3.06; -3.56, -2.56) was more than that related to solid fuel use for cooking (-1.28; -2.61, 0.05) and heating (-1.90; -3.16, -0.65). Similar results were observed in women but with a smaller magnitude. In this Chinese population, the harm to LE from household use of solid fuels was greater than that from smoking in rural residents. Conversely, the negative impact of smoking was greater than solid fuel use in urban residents. Our findings highlight the complexity and diversity of the factors affecting LE in less-developed populations. National Natural Science Foundation of China, National Key R&D Program of China, Kadoorie Charitable Foundation, UK Wellcome Trust.


Organizational Readiness for Intimate Partner Violence Response among Supportive Housing Providers: A Capacity Assessment in Maryland.
Intimate partner violence (IPV) frequently leads to housing instability and homelessness among survivors. While the client populations of many housing support programs are likely to include IPV survivors who have unique safety needs, the organizational readiness of these housing providers to identify and support IPV survivors is not clear. This study assessed organizational readiness for IPV response among Rapid Re-Housing (RRH) and Transitional Housing (TH) providers in Maryland, whose client populations include women (n = 32). We adapted the Physician Readiness to Manage Intimate Partner Violence Survey (PREMIS) to create the Housing provider Readiness to Manage IPV Survey (H-REMIS), inclusive of IPV-related perceptions, policies, staff training and capability, and resources. The 12-point H-REMIS demonstrated acceptable internal consistency reliability (Cronbach's α = 0.748). Descriptive and comparative analyses domains by IPV-specific (n = 4) and general (n = 28) housing providers assessed readiness and identified areas for improvement. IPV-specific providers had higher mean organizational readiness for IPV response scores relative to non-IPV specific providers (11.3 and 7.5, respectively). High readiness areas included perceiving the importance of IPV and staff having adequate time, space, and comfort level to address IPV. Areas of low readiness included development and use of IPV response policies as well as staff training on IPV. This assessment documents concerning gaps in organizational readiness for IPV response among supportive housing providers, and pinpoints areas where training and capacity building can be most valuable. The process of improving readiness in the supportive housing sector must involve capacity building and a systems-level approach in order to ensure that all supportive housing providers are prepared to meet the needs of IPV survivors among their client population.


Characterization of the folate salvage enzyme p-aminobenzoylglutamate hydrolase in plants.
Folates break down in vivo to give pterin and p-aminobenzoylglutamate (pABAGlu) fragments, the latter usually having a polyglutamyl tail. Pilot studies have shown that plants can hydrolyze pABAGlu and its polyglutamates to p-aminobenzoate, a folate biosynthesis precursor. The enzymatic basis of this hydrolysis was further investigated. pABAGlu hydrolase activity was found in all species and organs tested; activity levels implied that the proteins responsible are very rare. The activity was located in cytosol/vacuole and mitochondrial fractions of pea (Pisum sativum L.) leaves, and column chromatography of the activity from Arabidopsis tissues indicated at least three peaks. A major activity peak from Arabidopsis roots was purified 86-fold by a three-column procedure; activity loss during purification exceeded 95%. Size exclusion chromatography gave a molecular mass of approximately 200 kDa. Partially purified preparations showed a pH optimum near 7.5, a Km value for pABAGlu of 370 microM, and activity against folic acid. Activity was relatively insensitive to thiol and serine reagents, but was strongly inhibited by 8-hydroxyquinoline-5-sulfonic acid and stimulated by Mn2+, pointing to a metalloenzyme. The Arabidopsis genome was searched for proteins similar to Pseudomonas carboxypeptidase G, which contains zinc and is the only enzyme yet confirmed to attack pABAGlu. The sole significant matches were auxin conjugate hydrolase family members and the At4g17830 protein. None was found to have significant pABAGlu hydrolase activity, suggesting that this activity resides in hitherto unrecognized enzymes. The finding that Arabidopsis has folate-hydrolyzing activity points to an enzymatic component of folate degradation in plants.


Arginase-1: a new immunohistochemical marker of hepatocytes and hepatocellular neoplasms.
The distinction of hepatocellular carcinoma (HCC) from metastatic tumor in the liver often presents a diagnostic challenge that carries significant impact on prognostication and therapy. The number of diagnostically useful immunohistochemical markers of hepatocytes is limited to hepatocyte paraffin antigen (HepPar-1), polyclonal carcinoembryonic antigen, and CD10, with alpha-fetoprotein and glypican-3 labeling HCCs. Arginase-1 (Arg-1) is a binuclear manganese metalloenzyme that catalyzes the hydrolysis of arginine to ornithine and urea. We used immunohistochemistry to compare the sensitivity of Arg-1 to that of HepPar-1 in 151 HCCs. We found that the overall sensitivities of Arg-1 and HepPar-1 are 96.0% and 84.1%, respectively. The sensitivities of Arg-1 in well, moderately, and poorly differentiated HCCs are 100%, 96.2%, and 85.7%, respectively, whereas, in comparison, HepPar-1 demonstrated sensitivities of 100%, 83.0%, and 46.4% for well, moderately, and poorly differentiated tumors, respectively. There were no HCCs in our study that were reactive for HepPar-1 but nonreactive for Arg-1. We also examined Arg-1 expression in nonhepatocellular tumors, including many that are potential mimics of HCC (renal cell carcinomas, neuroendocrine tumors, melanomas, gastric adenocarcinomas, and adrenocortical carcinomas) and found that only 2 non-HCC tumors were reactive for Arg-1. Arg-1 represents a sensitive and specific marker of benign and malignant hepatocytes that may ultimately prove to be a useful diagnostic tool in routine surgical pathology practice.


A remarkable age-related increase in SIRT1 protein expression against oxidative stress in elderly: SIRT1 gene variants and longevity in human.
Aging is defined as the accumulation of progressive organ dysfunction. Controlling the rate of aging by clarifying the complex pathways has a significant clinical importance. Nowadays, sirtuins have become famous molecules for slowing aging and decreasing age-related disorders. In the present study, we analyzed the SIRT1 gene polymorphisms (rs7895833 A>G, rs7069102 C>G and rs2273773 C>T) and its relation with levels of SIRT1, eNOS, PON-1, cholesterol, TAS, TOS, and OSI to demonstrate the association between genetic variation in SIRT1 and phenotype at different ages in humans. We observed a significant increase in the SIRT1 level in older people and found a significant positive correlation between SIRT1 level and age in the overall studied population. The oldest people carrying AG genotypes for rs7895833 have the highest SIRT1 level suggesting an association between rs7895833 SNP and lifespan longevity. Older people have lower PON-1 levels than those of adults and children which may explain the high levels of SIRT1 protein as a compensatory mechanism for oxidative stress in the elderly. The eNOS protein level was significantly decreased in older people as compared to adults. There was no significant difference in the eNOS level between older people and children. The current study is the first to demonstrate age-related changes in SIRT1 levels in humans and it is important for a much better molecular understanding of the role of the longevity gene SIRT1 and its protein product in aging. It is also the first study presenting the association between SIRT1 expression in older people and rs7895833 in SIRT1 gene.


[Behavioral group-based therapy improves quality of life in men recovering from radical prostatectomy].
This study evaluated the efficacy of 24-week, group-based, behavioral therapy in improving quality of life (QoL) among men treated for localized prostate carcinoma (PC) by means of radical prostatectomy (RP) who presented treatment-related dysfunctions: urinary incontinence (UI) and erectile dysfunction (ED). Thirty men were assigned to 24-week behavioral group therapy. The intervention was designed to improve QoL by helping participants to identify and effectively manage stressful experiences. It focused on treatment-related sequelae from PC. Differences in variables were compared between the beginning and end of the study by means of Student's t test for paired samples. Multiple analysis was carried out by stepwise multiple linear regression following bivariate Pearson's correlation analysis. This was achieved for all predictors (i.e. general health perception, ED and UI impact) and relevant covariates (i.e. age, work/retirement status, alcohol addiction, attitude towards cancer and surgery, sexual satisfaction, and future plans). In all patients the time elapsed from surgery to attending the behavioral group therapy exceeded 24 months. There was no significant correlation between this time and the predictor variables. General health perception scores decreased by the end of the study (p = 0.000), as did the UI impact score (p = 0.023), thus denoting improvement in both factors. The difference in UI impact scores correlated negatively and significantly with both age (p = 0.04) and work/retirement (p = 0.05). Multiple stepwise regression showed that age was the most important variable (r2 = 26.0%). Considering age and work/retirement simultaneously, there was an increase of 10.3% (r2 = 36.3%). ED increased by the end of the study (p = 0.000), and the difference between the ED scores correlated positively and significantly with sexual satisfaction alone (p = 0.029), which signifies that previous sexual satisfaction had a positive influence over erectile dysfunction (r2 = 15.8%). 24-week behavioral group therapy was effective in improving the perceived QoL among men treated for PC. There were changes associated with the therapy, particularly the improvement in UI and ED.