Different models for prediction of radical cystectomy postoperative complications and care pathways.
Radical cystectomy for bladder cancer has one of the highest rates of morbidity among urologic surgery, but the ability to predict postoperative complications remains poor. Our study objective was to create machine learning models to predict complications and factors leading to extended length of hospital stay and discharge to a higher level of care after radical cystectomy. Using the American College of Surgeons National Surgical Quality Improvement Program, peri-operative adverse outcome variables for patients undergoing elective radical cystectomy for bladder cancer from 2005 to 2016 were extracted. Variables assessed include occurrence of minor, infectious, serious, or any adverse events, extended length of hospital stay, and discharge to higher-level care. To develop predictive models of radical cystectomy complications, we fit generalized additive model (GAM), least absolute shrinkage and selection operator (LASSO) logistic, neural network, and random forest models to training data using various candidate predictor variables. Each model was evaluated on the test data using receiver operating characteristic curves. A total of 7557 patients were identified who met the inclusion criteria, and 2221 complications occurred. LASSO logistic models demonstrated the highest area under curve for predicting any complications (0.63), discharge to a higher level of care (0.75), extended length of stay (0.68), and infectious (0.62) adverse events. This was comparable with random forest in predicting minor (0.60) and serious (0.63) adverse events. Our models perform modestly in predicting radical cystectomy complications, highlighting both the complex cystectomy process and the limitations of large healthcare datasets. Identifying the most important variable leading to each type of adverse event may allow for further strategies to model cystectomy complications and target optimization of modifiable variables pre-operative to reduce postoperative adverse events.


Extracellular Matrix Remodeling Regulates Glucose Metabolism through TXNIP Destabilization.
The metabolic state of a cell is influenced by cell-extrinsic factors, including nutrient availability and growth factor signaling. Here, we present extracellular matrix (ECM) remodeling as another fundamental node of cell-extrinsic metabolic regulation. Unbiased analysis of glycolytic drivers identified the hyaluronan-mediated motility receptor as being among the most highly correlated with glycolysis in cancer. Confirming a mechanistic link between the ECM component hyaluronan and metabolism, treatment of cells and xenografts with hyaluronidase triggers a robust increase in glycolysis. This is largely achieved through rapid receptor tyrosine kinase-mediated induction of the mRNA decay factor ZFP36, which targets TXNIP transcripts for degradation. Because TXNIP promotes internalization of the glucose transporter GLUT1, its acute decline enriches GLUT1 at the plasma membrane. Functionally, induction of glycolysis by hyaluronidase is required for concomitant acceleration of cell migration. This interconnection between ECM remodeling and metabolism is exhibited in dynamic tissue states, including tumorigenesis and embryogenesis.


Unravelling the origins of ice nucleation on organic crystals.
Organic molecules such as steroids or amino acids form crystals that can facilitate the formation of ice - arguably the most important phase transition on earth. However, the origin of the ice nucleating ability of organic crystals is still largely unknown. Here, we combine experiments and simulations to unravel the microscopic details of ice formation on cholesterol, a prototypical organic crystal widely used in cryopreservation. We find that cholesterol - which is also a substantial component of cell membranes - is an ice nucleating agent more potent than many inorganic substrates, including the mineral feldspar (one of the most active ice nucleating materials in the atmosphere). Scanning electron microscopy measurements reveal a variety of morphological features on the surfaces of cholesterol crystals: this suggests that the topography of the surface is key to the broad range of ice nucleating activity observed (from -4 to -20 °C). In addition, we show via molecular simulations that cholesterol crystals aid the formation of ice nuclei in a unconventional fashion. Rather than providing a template for a flat ice-like contact layer (as found in the case of many inorganic substrates), the flexibility of the cholesterol surface and its low density of hydrophilic functional groups leads to the formation of molecular cages involving both water molecules and terminal hydroxyl groups of the cholesterol surface. These cages are made of 6- and, surprisingly, 5-membered hydrogen bonded rings of water and hydroxyl groups that favour the nucleation of hexagonal as well as cubic ice (a rare occurrence). We argue that the phenomenal ice nucleating activity of steroids such as cholesterol (and potentially of many other organic crystals) is due to (i) the ability of flexible hydrophilic surfaces to form unconventional ice-templating structures and (ii) the different nucleation sites offered by the diverse topography of the crystalline surfaces. These findings clarify how exactly organic crystals promote the formation of ice, thus paving the way toward deeper understanding of ice formation in soft and biological matter - with obvious reverberations on atmospheric science and cryobiology.


Reconsideration of insulin signals induced by improved laboratory animal diets, Japanese and American diets, in IRS-2 deficient mice.
Current Japanese and American diets and Japanese diet immediately after the War were converted to laboratory animal diets. As a result, current laboratory animal diet (CA-1, CLEA) unexpectedly resembled the diet of Japanese after the War. This is considered to result in an under-evaluation of diabetes research using laboratory animals at present. Therefore, changes in insulin signals caused by current Japanese and American diets were examined using IRS-2 deficient mice ( IRS2(-/-) mice) and mechanisms of aggravation of type 2 diabetes due to modern diets were examined. IRS2(-/-) mice at 6 weeks of age were divided into three groups: Japanese diet (Jd) group, American diet (Ad) group and CA-1 diet [regular diet (Rd)] group. Each diet was given to the dams from 7 days before delivery. When the IRS2(-/-) mice reached 6 weeks of age, the glucose tolerance test (GTT), insulin tolerance test (ITT) and organ sampling were performed. The sampled organs and white adipose tissue were used for analysis of RNA, enzyme activity and tissues. In GTT and ITT, the Ad group showed worse glucose tolerance and insulin resistance than the Rd group. Impaired glucose tolerance of the Jd group was the same as that of the Rd group, but insulin resistance was worse than in the Rd group. These results were caused an increase in fat accumulation and adipocytes in the peritoneal cavity by lipogenic enzyme activity in the liver and muscle, and the increase in TNFalpha of hypertrophic adipocyte origin further aggravated insulin resistance and the increase in resistin also aggravated the impaired glucose tolerance, leading to aggravation of type 2 diabetes. The Japanese and American diets given to the IRS2(-/-) mice, which we developed, showed abnormal findings in some IRS2(-/-) mice but inhibited excessive reactions of insulin signals as diets used in ordinary nutritional management.


Atlas - a data warehouse for integrative bioinformatics.
We present a biological data warehouse called Atlas that locally stores and integrates biological sequences, molecular interactions, homology information, functional annotations of genes, and biological ontologies. The goal of the system is to provide data, as well as a software infrastructure for bioinformatics research and development. The Atlas system is based on relational data models that we developed for each of the source data types. Data stored within these relational models are managed through Structured Query Language (SQL) calls that are implemented in a set of Application Programming Interfaces (APIs). The APIs include three languages: C++, Java, and Perl. The methods in these API libraries are used to construct a set of loader applications, which parse and load the source datasets into the Atlas database, and a set of toolbox applications which facilitate data retrieval. Atlas stores and integrates local instances of GenBank, RefSeq, UniProt, Human Protein Reference Database (HPRD), Biomolecular Interaction Network Database (BIND), Database of Interacting Proteins (DIP), Molecular Interactions Database (MINT), IntAct, NCBI Taxonomy, Gene Ontology (GO), Online Mendelian Inheritance in Man (OMIM), LocusLink, Entrez Gene and HomoloGene. The retrieval APIs and toolbox applications are critical components that offer end-users flexible, easy, integrated access to this data. We present use cases that use Atlas to integrate these sources for genome annotation, inference of molecular interactions across species, and gene-disease associations. The Atlas biological data warehouse serves as data infrastructure for bioinformatics research and development. It forms the backbone of the research activities in our laboratory and facilitates the integration of disparate, heterogeneous biological sources of data enabling new scientific inferences. Atlas achieves integration of diverse data sets at two levels. First, Atlas stores data of similar types using common data models, enforcing the relationships between data types. Second, integration is achieved through a combination of APIs, ontology, and tools. The Atlas software is freely available under the GNU General Public License at: http://bioinformatics.ubc.ca/atlas/


Psychometric properties of the five facet mindfulness questionnaire in depressed adults and development of a short form.
In recent years, there has been a growing interest in therapies that include the learning of mindfulness skills. The 39-item Five Facet Mindfulness Questionnaire (FFMQ) has been developed as a reliable and valid comprehensive instrument for assessing different aspects of mindfulness in community and student samples. In this study, the psychometric properties of the Dutch FFMQ were assessed in a sample of 376 adults with clinically relevant symptoms of depression and anxiety. Construct validity was examined with confirmatory factor analyses and by relating the FFMQ to measures of psychological symptoms, well-being, experiential avoidance, and the personality factors neuroticism and openness to experience. In addition, a 24-item short form of the FFMQ (FFMQ-SF) was developed and assessed in the same sample and cross-validated in an independent sample of patients with fibromyalgia. Confirmatory factor analyses showed acceptable model fit for a correlated five-factor structure of the FFMQ and good model fit for the structure of the FFMQ-SF. The replicability of the five-factor structure of the FFMQ-SF was confirmed in the fibromyalgia sample. Both instruments proved highly sensitive to change. It is concluded that both the FFMQ and the FFMQ-SF are reliable and valid instruments for use in adults with clinically relevant symptoms of depression and anxiety.


High power all fiber mid-IR supercontinuum generation in a ZBLAN fiber pumped by a 2 μm MOPA system.
High power all fiber mid-IR supercontinuum (SC) generation in a ZBLAN fiber pumped by a 2 μm master oscillator power amplifier (MOPA) system is demonstrated. A semiconductor saturable absorber mirror (SESAM) passively mode-locked laser with pulse width of 26 ps at 1960 nm is used as the seed of the MOPA system. A laser spectrum extending from ~1.9 μm to beyond 2.6 μm is generated in a subsequent thulium-doped fiber amplifier (TDFA). Then, the spectrum is further broadened to the mid-IR region in the ZBLAN fiber. A mid-IR SC extending from 1.9 to 3.9 μm with 7.11 W average output power is obtained based on a large mode area TDFA, the SC power for wavelengths longer than 2.5 μm is 3.52 W with a power ratio of 49.5% with respect to the total SC power. The overall optical conversion efficiency from the 790 nm pump of the large mode area TDFA to the total SC output is 10.4%. To the best of our knowledge, both the 7.11 W total average power and 3.52 W average power in wavelengths beyond 2.5 μm are the highest power ever reported for a mid-IR SC generation in ZBLAN fiber pumped by 2 μm fiber lasers and TDFAs.


Multiplex immunoassay for Lyme disease using VlsE1-IgG and pepC10-IgM antibodies: improving test performance through bioinformatics.
The Centers for Disease Control and Prevention currently recommends a 2-tier serologic approach to Lyme disease laboratory diagnosis, comprised of an initial serum enzyme immunoassay (EIA) for antibody to Borrelia burgdorferi followed by supplementary IgG and IgM Western blotting of EIA-positive or -equivocal samples. Western blot accuracy is limited by subjective interpretation of weakly positive bands, false-positive IgM immunoblots, and low sensitivity for detection of early disease. We developed an objective alternative second-tier immunoassay using a multiplex microsphere system that measures VlsE1-IgG and pepC10-IgM antibodies simultaneously in the same sample. Our study population comprised 79 patients with early acute Lyme disease, 82 patients with early-convalescent-phase disease, 47 patients with stage II and III disease, 34 patients post-antibiotic treatment, and 794 controls. A bioinformatic technique called partial receiver-operator characteristic (ROC) regression was used to combine individual antibody levels into a single diagnostic score with a single cutoff; this technique enhances test performance when a high specificity is required (e.g., ≥ 95%). Compared to Western blotting, the multiplex assay was equally specific (95.6%) but 20.7% more sensitive for early-convalescent-phase disease (89.0% versus 68.3%, respectively; 95% confidence interval [95% CI] for difference, 12.1% to 30.9%) and 12.5% more sensitive overall (75.0% versus 62.5%, respectively; 95% CI for difference, 8.1% to 17.1%). As a second-tier test, a multiplex assay for VlsE1-IgG and pepC10-IgM antibodies performed as well as or better than Western blotting for Lyme disease diagnosis. Prospective validation studies appear to be warranted.


Sexual mixing patterns and sex-differentials in teenage exposure to HIV infection in rural Zimbabwe.
HIV-1 prevalence typically rises more rapidly at young ages in women than in men in sub-Saharan Africa. Greater susceptibility to infection on exposure in women is believed to be a contributory factor as is greater exposure to previously infected sexual partners of the opposite sex. We investigated the latter hypothesis using data from a field study in rural Manicaland, Zimbabwe. Quantitative data on onset and degree of sexual activity, numbers of partners, concurrent partnerships, condom use, and partner characteristics were used in conjunction with epidemiological data on age and sex specific prevalence of HIV infection to do statistical analyses of association between key variables. Mathematical models and qualitative data were used to aid analysis and interpretation. Older age of sexual partner was associated with increased risk of HIV-1 infection in men (odds ratio 1.13 [95% CI 1.02-1.25]) and women (1.04 [1.01-1.07]). Young women form partnerships with men 5-10 years older than themselves, whereas young men have relationships with women of a similar age or slightly younger. Greater number of lifetime partners is also associated with increased risk of HIV (1.03 [1.00-1.05]). Young men report more partners than do women but infrequent coital acts and greater use of condoms. These behaviour patterns are underpinned by cultural factors including the expectation that women should marry earlier than men. A strong gender effect remains after factors that affect exposure to infected partners are controlled for (6.04 [1.49-24.47]). The substantial age difference between female and male sexual partners in Manicaland is the major behavioural determinant of the more rapid rise in HIV prevalence in young women than in men. Theoretical studies have suggested that this difference is an important determinant of observed epidemiological patterns but the study reported in this paper provides clear empirical evidence of association.


Tandem fusion of hepatitis B core antigen allows assembly of virus-like particles in bacteria and plants with enhanced capacity to accommodate foreign proteins.
The core protein of the hepatitis B virus, HBcAg, assembles into highly immunogenic virus-like particles (HBc VLPs) when expressed in a variety of heterologous systems. Specifically, the major insertion region (MIR) on the HBcAg protein allows the insertion of foreign sequences, which are then exposed on the tips of surface spike structures on the outside of the assembled particle. Here, we present a novel strategy which aids the display of whole proteins on the surface of HBc particles. This strategy, named tandem core, is based on the production of the HBcAg dimer as a single polypeptide chain by tandem fusion of two HBcAg open reading frames. This allows the insertion of large heterologous sequences in only one of the two MIRs in each spike, without compromising VLP formation. We present the use of tandem core technology in both plant and bacterial expression systems. The results show that tandem core particles can be produced with unmodified MIRs, or with one MIR in each tandem dimer modified to contain the entire sequence of GFP or of a camelid nanobody. Both inserted proteins are correctly folded and the nanobody fused to the surface of the tandem core particle (which we name tandibody) retains the ability to bind to its cognate antigen. This technology paves the way for the display of natively folded proteins on the surface of HBc particles either through direct fusion or through non-covalent attachment via a nanobody.


Differential expression of miR-34b and androgen receptor pathway regulate prostate cancer aggressiveness between African-Americans and Caucasians.
African-Americans are diagnosed with more aggressive prostate cancers and have worse survival than Caucasians, however a comprehensive understanding of this health disparity remains unclear. To clarify the mechanisms leading to this disparity, we analyzed the potential involvement of miR-34b expression in African-Americans and Caucasians. miR-34b functions as a tumor suppressor and has a multi-functional role, through regulation of cell proliferation, cell cycle and apoptosis. We found that miR-34b expression is lower in human prostate cancer tissues from African-Americans compared to Caucasians. DNA hypermethylation of the miR-34b-3p promoter region showed significantly higher methylation in prostate cancer compared to normal samples. We then sequenced the promoter region of miR-34b-3p and found a chromosomal deletion in miR-34b in African-American prostate cancer cell line (MDA-PCA-2b) and not in Caucasian cell line (DU-145). We found that AR and ETV1 genes are differentially expressed in MDA-PCa-2b and DU-145 cells after overexpression of miR-34b. Direct interaction of miR-34b with the 3' untranslated region of AR and ETV1 was validated by luciferase reporter assay. We found that miR-34b downregulation in African-Americans is inversely correlated with high AR levels that lead to increased cell proliferation. Overexpression of miR-34b in cell lines showed higher inhibition of cell proliferation, apoptosis and G1 arrest in the African-American cells (MDA-PCa-2b) compared to Caucasian cell line (DU-145). Taken together, our results show that differential expression of miR-34b and AR are associated with prostate cancer aggressiveness in African-Americans.


The cycle of violence.
Despite widespread belief that violence begets violence, methodological problems substantially restrict knowledge of the long-term consequences of childhood victimization. Empirical evidence for this cycle of violence has been examined. Findings from a cohort study show that being abused or neglected as a child increases one's risk for delinquency, adult criminal behavior, and violent criminal behavior. However, the majority of abused and neglected children do not become delinquent, criminal, or violent. Caveats in interpreting these findings and their implications are discussed in this article.


Metabolic syndrome: do clinical criteria identify similar individuals among overweight premenopausal women?
The purpose of this analysis was to determine to what extent the clinical criteria for metabolic syndrome (MetSyn) proposed by the World Health Organization (WHO), the European Group for Study of Insulin Resistance (EGIR), the National Cholesterol Education Program Adult Treatment Panel III (ATP III), and the International Diabetes Foundation (IDF); triglyceride (TG)/high-density lipoprotein cholesterol (HDL-C) ratio >/=3.0; and enlarged waist circumference (>/=88 cm) and elevated TG (>/=129 mg/dL) (EWET) identified similar or different overweight women and, secondarily, to examine the effect of 7% weight reduction on MetSyn status. Metabolic syndrome was determined among 256 premenopausal women (age = 41 +/- 6 years, body mass index [BMI] = 32 +/- 4 kg/m(2)) participating in a dietary weight loss clinical trial based on the clinical criteria proposed by WHO, EGIR, ATP III, and IDF. The prevalence of TG/HDL-C ratio >/=3.0 and EWET was determined and compared with MetSyn status. Based on the clinical criteria, 16.1% (EGIR), 20.7% (WHO), 31.0% (ATP III), and 31.8% (IDF) of participants met the criteria for MetSyn; 30.3% and 31.8% had TG/HDL-C >/=3.0 and EWET, respectively. Between 77% and 99% of participants were similarly classified across the clinical criteria. The highest and lowest agreements were between ATP III and IDF (kappa = 0.98; 95% confidence interval, 0.96-1.0) and WHO and IDF (kappa = 0.39; 95% confidence interval, 0.26-0.51), respectively. The TG/HDL-C ratio >/=3.0 and EWET moderately agreed with all 4 clinical criteria for MetSyn (kappa range, 0.36-0.59). Among those diagnosed with MetSyn at baseline, 64.0% to 75.0% of the participants who lost >/=7% and 25.8% to 55.6% of participants who lost <7% of their baseline body weight in 6 months no longer met the various clinical criteria for MetSyn, TG/HDL-C >/=3.0, or EWET. Our findings indicate that MetSyn varies substantially between clinical criteria, which raise questions about the clinical utility of these criteria. Regardless of MetSyn clinical criteria, >/=7% reduction in body weight has a beneficial impact on variables used to define MetSyn.


Abnormalities in cartilage and bone development in the Apert syndrome FGFR2(+/S252W) mouse.
Apert syndrome is an autosomal dominant disorder characterized by malformations of the skull, limbs and viscera. Two-thirds of affected individuals have a S252W mutation in fibroblast growth factor receptor 2 (FGFR2). To study the pathogenesis of this condition, we generated a knock-in mouse model with this mutation. The Fgfr2(+/S252W) mutant mice have abnormalities of the skeleton, as well as of other organs including the brain, thymus, lungs, heart and intestines. In the mutant neurocranium, we found a midline sutural defect and craniosynostosis with abnormal osteoblastic proliferation and differentiation. We noted ectopic cartilage at the midline sagittal suture, and cartilage abnormalities in the basicranium, nasal turbinates and trachea. In addition, from the mutant long bones, in vitro cell cultures grown in osteogenic medium revealed chondrocytes, which were absent in the controls. Our results suggest that altered cartilage and bone development play a significant role in the pathogenesis of the Apert syndrome phenotype.


Identification of the delayed rectifier potassium channel, Kv1.6, in cultured astrocytes.
Astrocytes are an abundant glial cell type of the central nervous system that appear to play a role in regulating extracellular potassium concentrations in brain, thereby contributing to the maintenance of normal neuronal activity. Voltage-gated potassium conductances, shown to be present in astrocytes, may be involved in this and other astrocytic functions. Toward defining the role of voltage-gated potassium channels in astrocytes, total RNA prepared from cultured mouse cortical astrocytes was screened, using a reverse transcriptase-polymerase chain reaction (RT-PCR) approach, for the expression of several members of the Shaker-like potassium channel subfamily (Kv1.1-Kv1.6). A relatively high level of Kv1.6 transcript was identified by RT-PCR and then confirmed and quantitated by ribonuclease protection assays using a Kv1.6-specific riboprobe. Immunocytochemical staining showed double-labeling of glial fibrillary acidic protein-positive cells with antibody specific for the Kv1.6 channel. The Kv1.6 protein expression was variable among the individual astrocytes. Outward voltage-gated currents were studied in astrocytes in primary culture using the Nystatin-perforated patch voltage clamp technique. Outward potassium currents were observed in all cells studied, and this current was partially blocked by perfusion with 100 nM dendrotoxin (DTX) in 14 of 16 cells tested. This DTX-sensitive current appeared to be a sustained outward potassium current, consistent with the suggestion that the Shaker-like potassium channel Kv1.6 underlies a portion of the delayed rectifier potassium current in cultured mouse cortical astrocytes.


Tissue specific regulation of glucocorticoids in severe obesity and the response to significant weight loss following bariatric surgery (BARICORT).
Tissue cortisol exposure is under the control of the isozymes of 11β-hydroxysteroid dehydrogenase (11β-HSD). 11β-HSD1 in vivo, acts as an oxoreductase converting inactive cortisone to active cortisol. We hypothesized that 11β-HSD1 activity is dysregulated in obesity and alters following bariatric surgery induced weight loss in different tissues. We recruited 21 patients prior to undergoing bariatric surgery and performed cortisol generation profiles (following oral cortisone administration), urinary corticosteroid metabolite analysis, adipose tissue microdialysis, and tissue gene expression before and after weight loss, following bariatric surgery. Archived tissue samples from 20 previous bariatric surgery patients were also used for tissue gene expression studies. Gene expression showed a positive correlation with 11β-HSD1 and BMI in omental adipose tissue (OM) (r = +0.52, P = .0001) but not sc adipose tissue (r = +0.28, P = .17). 11β-HSD1 expression in liver negatively correlated with body mass index (BMI) (r = -0.37, P = .04). 11β-HSD1 expression in sc adipose tissue was significantly reduced after weight loss (0.41 ± 0.28 vs 0.17 ± 0.1 arbitrary units, P = .02). Following weight loss, serum cortisol generation increased during a cortisol generation profile (area under the curve 26 768 ± 16 880 vs 47 579 ± 16 086 nmol/L/minute, P ≤ .0001.) Urinary corticosteroid metabolites demonstrated a significant reduction in total cortisol metabolites after bariatric surgery (15 224 ± 6595 vs 8814 ± 4824 μg/24 h, P = .01). Microdialysis of sc adipose tissue showed a threefold reduction in cortisol/cortisone ratio after weight loss. This study highlights the differences in tissue specific regulation of cortisol metabolism in obesity and after weight loss. Following bariatric surgery hepatic 11β-HSD1 activity increases, sc adipose tissue 11β-HSD1 activity is reduced and total urinary cortisol metabolites are reduced indicating a possible reduction in hypothalamic pituitary adrenal axis drive. 11β-HSD1 expression correlates positively with BMI in omental adipose tissue and negatively within hepatic tissue. 11β-HSD1 expression is reduced in sc adipose tissue after weight loss.


In-bore setup and software for 3T MRI-guided transperineal prostate biopsy.
MRI-guided prostate biopsy in conventional closed-bore scanners requires transferring the patient outside the bore during needle insertion due to the constrained in-bore space, causing a safety hazard and limiting image feedback. To address this issue, we present our custom-made in-bore setup and software to support MRI-guided transperineal prostate biopsy in a wide-bore 3 T MRI scanner. The setup consists of a specially designed tabletop and a needle-guiding template with a Z-frame that gives a physician access to the perineum of the patient at the imaging position and allows the physician to perform MRI-guided transperineal biopsy without moving the patient out of the scanner. The software and Z-frame allow registration of the template, target planning and biopsy guidance. Initially, we performed phantom experiments to assess the accuracy of template registration and needle placement in a controlled environment. Subsequently, we embarked on our clinical trial (N = 10). The phantom experiments showed that the translational errors of the template registration along the right-left (RP) and anterior-posterior (AP) axes were 1.1 ± 0.8 and 1.4 ± 1.1 mm, respectively, while the rotational errors around the RL, AP and superior-inferior axes were (0.8 ± 1.0)°, (1.7 ± 1.6)° and (0.0 ± 0.0)°, respectively. The 2D root-mean-square (RMS) needle-placement error was 3 mm. The clinical biopsy procedures were safely carried out in all ten clinical cases with a needle-placement error of 5.4 mm (2D RMS). In conclusion, transperineal prostate biopsy in a wide-bore 3T scanner is feasible using our custom-made tabletop setup and software, which supports manual needle placement without moving the patient out of the magnet.


Oral microbiome activity in children with autism spectrum disorder.
Autism spectrum disorder (ASD) is associated with several oropharyngeal abnormalities, including buccal sensory sensitivity, taste and texture aversions, speech apraxia, and salivary transcriptome alterations. Furthermore, the oropharynx represents the sole entry point to the gastrointestinal (GI) tract. GI disturbances and alterations in the GI microbiome are established features of ASD, and may impact behavior through the "microbial-gut-brain axis." Most studies of the ASD microbiome have used fecal samples. Here, we identified changes in the salivary microbiome of children aged 2-6 years across three developmental profiles: ASD (n = 180), nonautistic developmental delay (DD; n = 60), and typically developing (TD; n = 106) children. After RNA extraction and shotgun sequencing, actively transcribing taxa were quantified and tested for differences between groups and within ASD endophenotypes. A total of 12 taxa were altered between the developmental groups and 28 taxa were identified that distinguished ASD patients with and without GI disturbance, providing further evidence for the role of the gut-brain axis in ASD. Group classification accuracy was visualized with receiver operating characteristic curves and validated using a 50/50 hold-out procedure. Five microbial ratios distinguished ASD from TD participants (79.5% accuracy), three distinguished ASD from DD (76.5%), and three distinguished ASD children with/without GI disturbance (85.7%). Taxonomic pathways were assessed using the Kyoto Encyclopedia of Genes and Genomes microbial database and compared with one-way analysis of variance, revealing significant differences within energy metabolism and lysine degradation. Together, these results indicate that GI microbiome disruption in ASD extends to the oropharynx, and suggests oral microbiome profiling as a potential tool to evaluate ASD status. Autism Res 2018, 11: 1286-1299. © 2018 International Society for Autism Research, Wiley Periodicals, Inc. LAY SUMMARY: Previous research suggests that the bacteria living in the human gut may influence autistic behavior. This study examined genetic activity of microbes living in the mouth of over 300 children. The microbes with differences in children with autism were involved in energy processing and showed potential for identifying autism status.


Peptidylarginine deiminase 2 has potential as both a biomarker and therapeutic target of sepsis.
Peptidylarginine deiminases (PADs) are a family of calcium-dependent enzymes that are involved in a variety of human disorders, including cancer and autoimmune diseases. Although targeting PAD4 has shown no benefit in sepsis, the role of PAD2 remains unknown. Here, we report that PAD2 is engaged in sepsis and sepsis-induced acute lung injury in both human patients and mice. Pad2-/- or selective inhibition of PAD2 by a small molecule inhibitor increased survival and improved overall outcomes in mouse models of sepsis. Pad2 deficiency decreased neutrophil extracellular trap (NET) formation. Importantly, Pad2 deficiency inhibited Caspase-11-dependent pyroptosis in vivo and in vitro. Suppression of PAD2 expression reduced inflammation and increased macrophage bactericidal activity. In contrast to Pad2-/-, Pad4 deficiency enhanced activation of Caspase-11-dependent pyroptosis in BM-derived macrophages and displayed no survival improvement in a mouse sepsis model. Collectively, our findings highlight the potential of PAD2 as an indicative marker and therapeutic target for sepsis.


Molecular and functional characterization of amylin, a peptide associated with type 2 diabetes mellitus.
The 37-amino acid peptide called amylin is a major component of the islet amyloid deposited in the pancreases of persons with type 2 diabetes mellitus. We report the isolation of a partial cDNA clone and a phage lambda genomic clone of the coding region of the amylin gene. The DNA sequence encodes a protein sequence identical to that of amylin isolated from the amyloid found in the diabetic pancreas and shows that amylin is likely to be synthesized as a precursor peptide, now named proamylin. We have demonstrated that the amylin gene is present on chromosome 12 and that it is probably transcribed in the islets of Langerhans. The sequences of the genes for amylin and the calcitonin gene-related peptides (CGRPs) show strong similarity, especially over their 5' coding regions, where both peptides have a conserved intramolecular disulfide bridge, and also over their 3' coding regions, where the presence of a glycine codon strongly suggests that the carboxyl-terminal residue of amylin, like that of CGRP, is amidated. To examine the functional relevance of these posttranslational modifications, the biological activity of amylin synthesized with or without the disulfide bridge and/or amidation was measured. It was found that both features are necessary for full biological activity, thereby confirming the functional importance of those regions of the molecule whose sequences are conserved at both protein and genetic levels.


Comparative evaluation of naftopidil and tamsulosin in the treatment of patients with lower urinary tract symptoms with benign prostatic hyperplasia.
Naftopidil, approved initially in Japan, is an α1d-adrenergic receptor antagonist (α1-blocker) used to treat lower urinary tract symptoms (LUTS) due to benign prostatic hyperplasia (BPH). It is different from tamsulosin hydrochloride and silodosin, in that it has a higher affinity for the α1D-adrenergic receptor subtype than for the α1A subtype and has a superior efficacy to a placebo and comparable efficacy to other α1-blockers such as tamsulosin. The incidences of ejaculatory disorders and intraoperative floppy iris syndrome induced by naftopidil may also be lower than that for tamsulosin and silodosin, which have a high affinity for the α1A-adrenergic receptor subtype. However, it remains unknown if the efficacy and safety of naftopidil in Japanese men is applicable to Indian men having LUTS/BPH. Two groups of 60 patients each, having LUTS due to BPH, were treated with tamsulosin 0.4 mg and Naftopidil 75 mg for three months. Ultrasonography (for prostate size, post-void residual volume), uroflowmetry, and the International Prostate Symptom Score (IPSS) and Quality of Life (QOL) score were recorded at the beginning of the study, and then at one and three months. The prostate size, post-void residual volume, all the uroflowmetry variables, and the IPSS QOL scores showed a statistically significant improvement (P < 0.001) in both the groups. The improvement in the average flow rate and the QOL index was better in the naftopidil group on the intergroup comparison and was statistically significant (P < 0.001). Although the QOL life index was significantly better in the naftopidil group, overall both naftopidil and tamsulosin were found to be equally effective in the treatment of LUTS due to BPH.


One-step multiplex quantitative RT-PCR for the simultaneous detection of viroids and phytoplasmas of pome fruit trees.
A one-step multiplex real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) based on TaqMan chemistry was developed for the simultaneous detection of Pear blister canker viroid and Apple scar skin viroid along with universal detection of phytoplasmas, in pome trees. Total nucleic acids (TNAs) extraction was performed according to a modified CTAB protocol. Primers and TaqMan MGB probes for specific detection of the two viroids were designed in this study, whereas for phytoplasma detection published universal primers and probe were used, with the difference that the later was modified to carry a MGB quencher. The pathogens were detected simultaneously in 10-fold serial dilutions of TNAs from infected plant material into TNAs of healthy plant up to dilutions 10(-5) for viroids and 10(-4) for phytoplasmas. The multiplex real-time assay was at least 10 times more sensitive than conventional protocols for viroid and phytoplasma detection. Simultaneous detection of the three targets was achieved in composite samples at least up to a ratio of 1:100 triple-infected to healthy tissue, demonstrating that the developed assay has the potential to be used for rapid and massive screening of viroids and phytoplasmas of pome fruit trees in the frame of certification schemes and surveys.


Rational selection of inhalation devices in the treatment of chronic obstructive pulmonary disease by means of the System of Objectified Judgement Analysis (SOJA).
The large number of available medicines and devices makes it almost impossible to have sufficient knowledge of each individual medicine and device, especially for general practitioners. This may lead to suboptimal treatment, more exacerbations, hospitalisations and higher treatment costs. Reducing the number of medicines and devices, based on rational criteria, allows physicians and pharmacists to build experience with a more limited set of medicines and to standardise the inhalation instructions. In this study inhalers are compared by means of the System of Objectified Judgement Analysis (SOJA) method. The following selection criteria were applied: uniformity in device, number of steps per inhalation, risk of errors, hygienic aspects, feedback mechanism, and risk of inhalation with an empty inhaler. Substantial differences were seen in the overall scores, with the Ellipta device showing the highest score, followed by Diskus/Accuhaler, Genuair and Nexthaler. Several devices require more or less identical techniques, such Ellipta and Diskus/Accuhaler as well as Genuair and Novolizer. When patients use these devices in combination this increases their uniformity, because additional medicines become available for the devices: starting therapy with Diskus or Novolizer and follow-up with Ellipta or Genuair. The resistance of Respimat and Breezhaler is lower than that of other devices, which makes these devices suitable for patients who cannot generate sufficient inhalation flow. A substantial reduction of inhalers, combined with optimal and standardised instructions, should improve the care of patients with chronic obstructive pulmonary disease.


Transcriptome analysis of genes regulated by cholesterol loading in two strains of mouse macrophages associates lysosome pathway and ER stress response with atherosclerosis susceptibility.
Cholesterol loaded macrophages in the arterial intima are the earliest histological evidence of atherosclerosis. Studies of mouse models of atherosclerosis have shown that the strain background can have a significant effect on lesion development. We have previously shown that DBA/2 ApoE(-/-) mice have aortic root lesions 10-fold larger than AKR ApoE(-/-) mice. The current study analyzes the response to cholesterol loading of macrophages from these two strains. Macrophages from the atherosclerosis susceptible DBA/2 strain had significantly higher levels of total and esterified cholesterol compared to atherosclerosis resistant AKR macrophages, while free cholesterol levels were higher in AKR cells. Gene expression profiles were obtained and data were analyzed for strain, cholesterol loading, and strain-cholesterol loading interaction effects by a fitted linear model. Pathway and transcriptional motif enrichment were identified by gene set enrichment analysis. In addition to observed strain differences in basal gene expression, we identified many transcripts whose expression was significantly altered in response to cholesterol loading, including P2ry13 and P2ry14, Trib3, Hyal1, Vegfa, Ccr5, Ly6a, and Ifit3. Eight pathways were significantly enriched in transcripts regulated by cholesterol loading, among which the lysosome and cytokine-cytokine receptor interaction pathways had the highest number of significantly regulated transcripts. Of the differentially regulated transcripts with a strain-cholesterol loading interaction effect, we identified three genes known to participate in the endoplasmic reticulum (ER) stress response, Ddit3, Trib3 and Atf4. These three transcripts were highly up-regulated by cholesterol in AKR and either down-regulated or unchanged in loaded DBA/2 macrophages, thus associating a robust ER stress response with atherosclerosis resistance. We identified significant transcripts with strain, loading, or strain-loading interaction effect that reside within previously described quantitative trait loci as atherosclerosis modifier candidate genes. In conclusion, we characterized several strain and cholesterol induced differences that may lead to new insights into cellular cholesterol metabolism and atherosclerosis.


The use of tritium content as an indicator of the groundwater contamination by sanitary landfills leachates in the region of Belo Horizonte City, Brazil.
Tritium content in the leachate of sanitary landfills, in concentrations well above those observed in global precipitation, can be used as a tracer for the evaluation of the contamination of groundwater in piezometers of the landfills and in neighbouring tubular wells. This possibility was first investigated in Brazil for sanitary landfills in the region of Belo Horizonte City. Tritium levels together with the content of metals present in water and the measurement of soil electrical conductivity, proved to be valuable for these studies and also as a tracer for hydrodynamic studies of the surface water in the Ressaca creek.


Effect of UVA-activated riboflavin on dentin bonding.
Recent studies have reported collagen cross-linking after exposure to riboflavin followed by ultraviolet-A (UVA) exposure. This study is the first to investigate the effect of a riboflavin-containing primer on adhesive interface stability and dentinal matrix metalloproteinase activity. Human dentin was etched with 35% phosphoric acid, treated with 0.1% riboflavin, exposed to UVA for 2 min, and bonded with a two-step etch-and-rinse adhesive. Adhesive was applied to control specimens without riboflavin/UVA. Specimens were subjected to microtensile bond strength tests and pulled to failure after storage for 24 hrs, 6 mos, or 1 yr. Interfacial nanoleakage was evaluated by light and transmission electron microscopy. To investigate dentinal matrix metalloproteinase activity, we performed correlative zymographic assays on protein extracts obtained from phosphoric-acid-etched dentin powder with or without riboflavin/UVA treatment and XP Bond. Ultraviolet-activated riboflavin treatment increased the immediate bond strength to dentin at all aging intervals (p < 0.05 vs. control) and decreased interfacial nanoleakage in aged specimens (1 yr; p < 0.05). Zymograms revealed that riboflavin/UVA pre-treatment inhibited dentinal matrix metalloproteinase activity (especially MMP-9). In conclusion, dentinal collagen cross-linking induced by riboflavin/UVA increased immediate bond strength, stabilized the adhesive interface, and inhibited dentin matrix metalloproteinases, thereby increasing the durability of resin-dentin bonds.


Inconsistency in Diarrhea Measurements when Assessing Intervention Impact in a Non-Blinded Cluster-Randomized Controlled Trial.
To explore the consistency in impact evaluation based on reported diarrhea, we compared diarrhea data collected through two different surveys and with observed diarrhea-associated hospitalization for children aged ≤ 5 years from a non-blinded cluster-randomized trial conducted over 2 years in urban Dhaka. We have previously reported that the interventions did not reduce diarrhea-associated hospitalization for children aged ≤ 5 years in this trial. We randomly allocated 90 geographic clusters comprising > 60,000 low-income households into three groups: cholera vaccine only, vaccine plus behavior change (cholera vaccine and handwashing plus drinking water chlorination promotion), and control. We calculated reported diarrhea prevalence within the last 2 days using data collected from two different survey methods. The "census" data were collected from each household every 6 months for updating household demographic information. The "monthly survey" data were collected every month from a subset of randomly selected study households for monitoring the uptake of behavior change interventions. We used binomial regression with a logarithmic link accounting for clustering to compare diarrhea prevalence across intervention and control groups separately for both census and monthly survey data. No intervention impact was detected in the census (vaccine only versus control: 2.32% versus 2.53%; P = 0.49; vaccine plus behavior change versus control: 2.44% versus 2.53%; P = 0.78) or in the vaccine only versus control in the monthly survey (3.39% versus 3.80%; P = 0.69). However, diarrhea prevalence was lower in the vaccine-plus-behavior-change group than control in the monthly survey (2.08% versus 3.80%; P = 0.02). Although the reasons for different observed treatment effects in the census and monthly survey data in this study are unclear, these findings emphasize the importance of assessing objective outcomes along with reported outcomes from non-blinded trials.


Transcriptome landscape of the human placenta.
The placenta is a key component in understanding the physiological processes involved in pregnancy. Characterizing genes critical for placental function can serve as a basis for identifying mechanisms underlying both normal and pathologic pregnancies. Detailing the placental tissue transcriptome could provide a valuable resource for genomic studies related to placental disease. We have conducted a deep RNA sequencing (RNA-Seq) study on three tissue components (amnion, chorion, and decidua) of 5 human placentas from normal term pregnancies. We compared the placental RNA-Seq data to that of 16 other human tissues and observed a wide spectrum of transcriptome differences both between placenta and other human tissues and between distinct compartments of the placenta. Exon-level analysis of the RNA-Seq data revealed a large number of exons with differential splicing activities between placenta and other tissues, and 79% (27 out of 34) of the events selected for RT-PCR test were validated. The master splicing regulator ESRP1 is expressed at a proportionately higher level in amnion compared to all other analyzed human tissues, and there is a significant enrichment of ESRP1-regulated exons with tissue-specific splicing activities in amnion. This suggests an important role of alternative splicing in regulating gene function and activity in specific placental compartments. Importantly, genes with differential expression or splicing in the placenta are significantly enriched for genes implicated in placental abnormalities and preterm birth. In addition, we identified 604-1007 novel transcripts and 494-585 novel exons expressed in each of the three placental compartments. Our data demonstrate unique aspects of gene expression and splicing in placental tissues that provide a basis for disease investigation related to disruption of these mechanisms. These data are publicly available providing the community with a rich resource for placental physiology and disease-related studies.


Postoperative Rehabilitation After Hip Resurfacing: A Systematic Review.
Hip-resurfacing arthroplasty (HRA) has become a popular procedure in the treatment of hip-joint arthritis in individuals under the age of 65 y. Although the body of literature examining operative procedures has grown, there is a lack of consistent reporting of the effectiveness of an HRA postoperative rehabilitation program. To date, no systematic reviews have evaluated the available evidence on postoperative rehabilitation programs. To evaluate the available evidence on postoperative rehabilitation programs after HRA. A systematic review was conducted according to the PRISMA guidelines. A search of PubMed, CINAHL, SPORTDiscus, ProQuest, and Google Scholar was conducted in April 2014 using the following keywords alone and in combination: postoperative, postsurgical, rehabilitation, physical therapy, programs, hip resurfacing, arthroplasty, and metal-on-metal. The grading of studies was conducted using the PEDro and Oxford Centre for Evidence-Based Medicine scales. The authors identified 648 citations, 4 of which met the inclusion criteria. The qualifying studies yielded 1 randomized control trial, 2 case reports, and 1 case series, for a total of 90 patients. Patients were mostly male (n = 86), had a mean age of 48 ± 5.47 y, and had been physically active before HRA. Postoperative rehabilitation programs varied in length (range 8-24 wk) and consisted of at least 3 phases. The methodology to assess program effects varied, but all 4 studies did measure a combination of function, pain, and quality of life using written questionnaires, with follow-up ranging from 9 mo to 1 y. The most common questionnaire was the Harris Hip Score. This review found postoperative rehabilitation programs after HRA to be underinvestigated. Limited results indicate that postoperative rehabilitation programs may be effective in improving gait (stride length, velocity, and cadence), hip range of motion, and pain and function, as measured by questionnaires, but not hip strength.


Base stacking and even/odd behavior of hairpin loops in DNA triplet repeat slippage and expansion with DNA polymerase.
Repetitions of CAG or CTG triplets in DNA can form intrastrand hairpin loops with combinations of normal and mismatched base pairs that easily rearrange. Such loops may promote primer-template slippage in DNA replication or repair to give triplet-repeat expansions like those associated with neurodegenerative diseases. Using self-priming sequences (e.g. (CAG)(16)(CTG)(4)), we resolve all hairpin loops formed and measure their slippage and expansion rates with DNA polymerase at 37 degrees C. Comparing CAG/CTG loop structures with GAC/GTC structures, having similar hydrogen bonding but different base stacking, we find that CAG, CTG, and GTC triplets predominantly form even-membered loops that slip in steps of two triplets, whereas GAC triplets favor odd-numbered loops. Slippage rates decline as hairpin stability increases, supporting the idea that slippage initiates more easily in less stable regions. Loop stabilities (in low salt) increase in the order GTC < CAG < GAC < CTG, while slippage rates decrease in the order GTC > CAG approximately GAC > CTG. Loops of GTC compared with CTG melt 9 degrees C lower and slip 6-fold faster. We interpret results in terms of base stacking, by relating melting temperature to standard enthalpy changes for doublets of base pairs and mispairs, considering enthalpy-entropy compensation.


Comparison of bacterial and dye microleakage of different root-end filling materials.
The provision of an effective apical seal is an essential factor when choosing an appropriate material for success of root-end surgeries. Root-end resection, preparation and obturation should provide an adequate apical seal. The aim of this in vitro study was to investigate bacterial and dye microleakage of four different root-end filling materials and compare the efficacy of these two methods. Fifty-six single-rooted teeth with intact and straight roots were randomly assigned into four study groups of 12 each and two control groups of three each. After decoronation, root canals were prepared up to file size #40 using step back technique; then, they were filled with gutta-percha and AH26 sealer. Root-ends were resected 3 mm above the root-end and 3 mm deep cavities were prepared. Root-end cavities were filled with amalgam, Root Mineral Trioxide Aggregate (Root MTA), White ProRoot MTA (WMTA), and calcium enriched mixture (CEM) cement. Bacterial leakage was investigated in Trypticase Soy Broth (containing Enterococcus faecalis) after 70 days and 1% methylene blue dye leakage was assessed after 72 hours. Complete dye leakage was checked using stereomicroscope (×40). Data were statistically analyzed using Fisher Exact test. For pair comparison between the two methods Kapa agreement was utilized. After 70 days there was 100% bacterial leakage in amalgam, and CEM cement, 91.7% in WMTA, and 75% in Root MTA. This difference was not significantly different. The difference in complete dye leakage was also not significant (WMTA and CEM cement 16.7%, Amalgam and Root MTA 33.3%). There was no significant measure of agreement between dye and bacterial penetration along root-end fillings. CEM cement was not significantly different from currently used retrofilling materials e.g. WMTA.


Temporal Confounding in the Test-Negative Design.
In the test-negative design, routine testing at health-care facilities is leveraged to estimate the effectiveness of an intervention such as a vaccine. The odds of vaccination for individuals who test positive for a target pathogen is compared with the odds of vaccination for individuals who test negative for that pathogen, adjusting for key confounders. The design is rapidly growing in popularity, but many open questions remain about its properties. In this paper, we examine temporal confounding by generalizing derivations to allow for time-varying vaccine status, including out-of-season controls, and open populations. We confirm that calendar time is an important confounder when vaccine status varies during the study. We demonstrate that, where time is not a confounder, including out-of-season controls can improve precision. We generalize these results to open populations. We use our theoretical findings to interpret 3 recent papers utilizing the test-negative design. Through careful examination of the theoretical properties of this study design, we provide key insights that can directly inform the implementation and analysis of future test-negative studies.


Parkinson's Is Time on Your Side? Evidence for Difficulties with Sensorimotor Synchronization.
There is lack of consistent evidence as to how well PD patients are able to accurately time their movements across space with an external acoustic signal. For years, research based on the finger-tapping paradigm, the most popular paradigm for exploring the brain's ability to time movement, has provided strong evidence that patients are not able to accurately reproduce an isochronous interval [i.e., Ref. (1)]. This was undermined by Spencer and Ivry (2) who suggested a specific deficit in temporal control linked to emergent, rhythmical movement not event-based actions, which primarily involve the cerebellum. In this study, we investigated motor timing of seven idiopathic PD participants in event-based sensorimotor synchronization task. Participants were asked to move their finger horizontally between two predefined target zones to synchronize with the occurrence of two sound events at two time intervals (1.5 and 2.5 s). The width of the targets and the distance between them were manipulated to investigate impact of accuracy demands and movement amplitude on timing performance. The results showed that participants with PD demonstrated specific difficulties when trying to accurately synchronize their movements to a beat. The extent to which their ability to synchronize movement was compromised was found to be related to the severity of PD, but independent of the spatial constraints of the task.


Improving engagement with services to prevent Sudden Unexpected Death in Infancy (SUDI) in families with children at risk of significant harm: A systematic review of evidence.
This paper reports part of a wider systematic review commissioned by the English National Safeguarding Panel on Sudden Unexpected Death in Infancy (SUDI). The wider review covered three areas: interventions to improve safer sleep practices in high-risk families, interventions to improve engagement with services and decision making by parents at high risk of SUDI about infant sleep environments. Here, we report the qualitative and quantitative studies reviewed under the engagement strand. Parental engagement is understood to be a multidimensional task for health and social care professionals comprising attitudinal, relational and behavioural components. Following a PROSPERO registered systematic review synthesizing the three strands outlined, 28 papers were found to be relevant in the review of interventions to improve engagement with services in families with children at risk of significant harm through abuse or neglect. No studies were found that specifically focused on engagement of families at high risk for SUDI, so these wider engagement studies were included. The different types of intervention reported in the included studies are described under two broad themes: Enablers (including parental motivation and working with families) and Barriers. Given the focus in the studies on interventions that support parental engagement, the Enablers theme is more extensive than the Barriers reported although all studies noted well-understood barriers. The evidence underpinning these interventions and approaches are reviewed in this paper. We conclude that effective engagement is facilitated by experienced professionals given time to develop supportive non-judgemental relationships with families in their homes, working long-term, linking with communities and other services. While these conclusions have been drawn from wider studies aimed at reducing child maltreatment, we emphasize lessons to be drawn for SUDI prevention work with families with children at risk of significant harm.


Polymorphisms of mtDNA control region in Tunisian and Moroccan populations: an enrichment of forensic mtDNA databases with Northern Africa data.
Current forensic mitochondrial (mt)DNA databases are limited in representative population data of African origin. We investigated HVS-I/HVS-II sequences of 120 Tunisian and Moroccan healthy male donors applying stringent quality criteria to assure high quality of the data and phylogenetic alignment and notation of the sequences. Among 64 Tunisians, 56 different haplotypes were observed and the most common haplotype (16187T 16189C 16223T 16264T 16270T 16278T 16293G 16311C 73G 152C 182T 185T 195C 247A 263G 309.1C 315.1C; haplogroup (hg) L1b) was shared by four individuals. 56 Moroccans could be assigned to 52 different haplotypes where the most common haplotype was of West Eurasian origin with the hg H sequence motif 263G 315.1C and variations in the HVS-II polyC-stretch (309.1C 309.2C) shared by six samples. The majority of the observed haplotypes belong to the west Eurasian phylogeny (50% in Tunisians and 62.5% in Moroccans). Our data are consistent with the current phylogeographic knowledge displaying the occurrence of sub-Saharan haplogroup L sequences, found in 48.4% of Tunisians and 25% of Moroccans as well as the presence of the two re-migrated haplogroups U6 (7.8% and 1.8% in Tunisians and Moroccans, respectively) and M1 (1.6% in Tunisians and 8.9% in Moroccans).


Substance use disorders in Iraq and Afghanistan veterans in VA healthcare, 2001-2010: Implications for screening, diagnosis and treatment.
The prevalence and correlates of alcohol use disorder (AUD) and drug use disorder (DUD) diagnoses in Iraq and Afghanistan veterans who are new users of Department of Veterans Affairs (VA) healthcare nationwide has not been evaluated. VA administrative data were used in retrospective cross-sectional descriptive and multivariable analyses to determine the prevalence and independent correlates of AUD and DUD in 456,502 Iraq and Afghanistan veterans who were first-time users of VA healthcare between October 15, 2001 and September 30, 2009 and followed through January 1, 2010. Over 11% received substance use disorder diagnoses: AUD, DUD or both; 10% received AUD diagnoses, 5% received DUD diagnoses and 3% received both. Male sex, age < 25 years, being never married or divorced, and proxies for greater combat exposure were independently associated with AUD and DUD diagnoses. Of those with AUD, DUD or both diagnoses, 55-75% also received PTSD or depression diagnoses. AUD, DUD or both diagnoses were 3-4.5 times more likely in veterans with PTSD and depression (p < 0.001). Post-deployment AUD and DUD diagnoses were more prevalent in subgroups of Iraq and Afghanistan veterans and were highly comorbid with PTSD and depression. Stigma and lack of universal screening may have reduced the number of DUD diagnoses reported. There is a need for improved screening and diagnosis of substance use disorders and increased availability of integrated treatments that simultaneously address AUD and DUD in the context of PTSD and other deployment-related mental health disorders.


First Report of Races 11 and 12 of Cercospora sojina, the Causal Agent of Soybean Frogeye Leaf Spot, in Argentina.
During the growing seasons of 2008 to 2009 and 2009 to 2010, severe outbreaks of soybean (Glycine max (L.) Merr.) frogeye leaf spot, a disease caused by Cercospora sojina Hara, occurred in several areas in Argentina (1). Two surveys were conducted in soybean fields, one in 2008 that included the provinces of Buenos Aires, Córdoba, and Santa Fe, and another that was performed in 2009 in the same provinces plus three others: Entre Ríos, Santiago del Estero, and Tucumán. In both surveys, plants presented circular lesions with reddish brown-to-gray spots and bordered by typical, narrow, reddish purple margins (3). To promote sporulation and to enable identification of the causal agent, leaves of diseased plants were collected and placed in a moist chamber for 24 h with a 12-h light cycle at 25°C. Conidia were plated on potato dextrose agar medium amended with streptomycin and were incubated at 25°C and 12 h of fluorescent light. Isolated cultures sporulated in 10 days and, on the basis of their morphology, were identified as C. sojina. A total of 147 isolates were deposited at the Culture Collection of CEREMIC (Centro de Referencia de Micología). They produced one- to nine-septate hyaline, elongate to fusiform conidia that measured 54.9 ± 16.2 × 5.7 ± 1.0 μm. Six isolates of C. sojina, each representing a province, were inoculated on a set of 12 differential soybean cultivars: Lee, Davis, Hood, Richland, Lincoln, Kent, Tracy, S 100, Palmetto, Peking, CNS, and Blackhawk (2). Fifteen plants of each differential were sprayed at V3 growth stage with a suspension of 6 × 104 conidia/ml. The test was conducted twice in a complete randomized design with three replicates. Control plants were sprayed with sterile distilled water. After inoculation, plants were placed in a greenhouse bench humidity chamber at 26 to 28°C for 72 h. Disease was rated 14 days after inoculation; plants with numerous lesions were considered susceptible and each of the 15 plants was given a score of 1. Plants with small or no lesions were classified as resistant and given a score of 0. Control plants remained healthy. The pathogen was reisolated from symptomatic plants and morphological characteristics were consistent with C. sojina. Based on the response of the differentials to each isolate and on the race designations, the isolates from Buenos Aires, Córdoba, Santa Fe, and Tucumán belong to race 11, while those from Santiago del Estero and Entre Ríos province to race 12. The finding of these two races threatening soybean cultivars in Argentina may be indicative of additional races. Thus, the incorporation of multiple resistance genes may reduce the impact of the disease on soybean. To our knowledge, this is the first report of the identification of races of C. sojina in Argentina. References: (1) M. A. Carmona et al. Plant Dis. 93:966, 2009. (2) M. A. R. Mian et al. Crop Sci. 48:14, 2008. (3) D.V. Phillips. Page 20 in: Compendium of Soybean Diseases. 4th ed. APS Press, St. Paul, MN, 1999.


Self-assembling small molecules for the detection of important analytes.
Nano-materials formed by the self-assembly of small molecules are very promising for drug delivery, regenerative medicine, and detection of important analytes due to their unique properties, such as self-assembled multivalency, biocompatibility, and fast response to external stimuli. This tutorial review focuses on their applications in detection of important analytes. Self-assembling small molecules can show fast response to external stimuli. Therefore, the gel-sol/sol-gel phase transitions of supramolecular hydrogels that can be easily identified by naked eyes have been applied for the detection of enzymes and enzyme-involving analytes. The supramolecular hydrogels can also provide semi-wet environments that can retain the activity of enzymes and recognition properties of molecular probes. Thus, they provide good platforms for the detection of many biologically and environmentally important analytes. Besides, self-assembling small molecules show big differences in fluorescence or the F-NMR signal between their self-assembled and un-assembled stages. Such small molecules can be rationally designed through the integration of fluorescent dyes or fluorine containing molecules in the self-assembling small molecules. Therefore, extensive recent research efforts have been made to explore their detection applications based on the dis-assembly triggered fluorescence/F-NMR signal turn on or the self-assembly/aggregation induced fluorescence turn on. We believe that the research efforts made to this field will ultimately lead to the development of useful nano-materials for detection applications.


Vascularized 3D printed scaffolds for promoting bone regeneration.
3D printed scaffolds hold promising perspective for bone tissue regeneration. Inspired by process of bone development stage, 3D printed scaffolds with rapid internal vascularization ability and robust osteoinduction bioactivity will be an ideal bone substitute for clinical use. Here, we fabricated a 3D printed biodegradable scaffold that can control release deferoxamine, via surface aminolysis and layer-by-layer assembly technique, which is essential for angiogenesis and osteogenesis and match to bone development and reconstruction. Our in vitro studies show that the scaffold significantly accelerates the vascular pattern formation of human umbilical endothelial cells, boosts the mineralized matrix production, and the expression of osteogenesis-related genes during osteogenic differentiation of mesenchymal stem cells. In vivo results show that deferoxamine promotes the vascular ingrowth and enhances the bone regeneration at the defect site in a rat large bone defect model. Moreover, this 3D-printed scaffold has excellent biocompatibility that is suitable for mesenchymal stem cells grow and differentiate and possess the appropriate mechanical property that is similar to natural cancellous bone. In summary, this 3D-printed scaffold holds huge potential for clinical translation in the treatment of segmental bone defect, due to its flexibility, economical friendly and practicality.


Neuraminidase-specific antibody responses are generated in naive and vaccinated newborn nonhuman primates following virus infection.
Individuals younger than 6 months of age are at significant risk from influenza virus infection; however, there is currently no vaccine approved for this age group. Influenza virus neuraminidase (NA) has emerged as a potential additional target for vaccine strategies. In this study, we sought to understand the ability of newborns to mount an antibody response to NA. Here we employed a nonhuman primate model, given the similarities to humans in immune system and development. We measured antibody to NA following infection with an H1N1 virus or following vaccination and challenge. Administration of an inactivated virus vaccine was not capable of eliciting detectable NA-specific antibody, even in the presence of adjuvants previously shown to increase total virus-specific IgG. However, both naive and vaccinated newborns generated a NA-specific antibody response following virus infection. Interestingly, the presence of the vaccine-induced response did not prevent generation of systemic antibody to NA following challenge, although the respiratory response was reduced in a significant portion of newborns. These findings are the first, to our knowledge, to evaluate the newborn response to the influenza NA protein as well as the impact of previous vaccination on generation of these antibodies following virus infection.


Predictors of Functional Improvements After Therapeutic Yoga Intervention for People with Parkinson's Disease.
Parkinson's disease (PD) affects nearly 10 million people worldwide, leading to decreased mobility and balance and potential loss of independence. Yoga has been associated with improved function for people with PD, but the predictive factors for improved functional outcomes as a result of yoga participation remain unexamined. The objective of this secondary data analysis was to identify predictive factors of functional improvement for people with PD after an 8-week yoga intervention. Stepwise multiple linear regression was used to identify significant predictors of improvement in balance, fall control, PD symptoms, and activity constraints. Lower cognitive functioning was predictive of improvement in perceived control over falls, body responsiveness was predictive of improvement in PD-specific symptoms, and gait velocity was predictive of improvement in balance and activity constraints. Future research is warranted to evaluate the use of screeners to predict who is the best fit for yoga therapy. Additional research is needed to evaluate the need to include cognitive self-management training concurrent with yoga therapy.


Cluster analysis of mRNA expression levels identifies multiple sequential patterns following focal cerebral ischemia.
The purpose of this study is to detect gene expression patterns following focal cerebral ischemia. 25 male Wistar rats were divided into control (n = 8) and ischemic (n = 17) groups. In the ischemic group, slowly progressing focal ischemia was simulated by two-vein occlusion with spreading depression (SD) a cortical microinjection of KCl induced. Ischemic tissue was removed at 2, 8, 24, or 72 h postischemia. Using semiquantitative reverse transcription polymerase chain reaction, we investigated mRNA expression levels of 13 representative genes related to cerebral ischemia. Cluster analysis of the gene expression levels was done. In the ischemic group, the expression levels of c-fos, cyclin D1, and COX-2 were significantly higher at 2 h postischemia, and those of bcl-2, bcl-xL, and HO-1 at 72 h. Based on the cluster analysis, we statistically divided examined genes into three groups: group A, early expression (COX-2, c-fos, and bcl-2); group B, s expression (c-jun, SOD-1, bad, p53, SOD-2, bcl-xL, and bax); and group C, late expression (cyclin D1, c-myc, and HO-1). We statistically classified the genes into three groups after focal ischemia. The genes of the early- and late-expression groups can be possible therapeutic targets for the treatment of cerebral ischemia.


Digital Books with Dynamic Text and Speech Output: Effects on Sight Word Reading for Preschoolers with Autism Spectrum Disorder.
Despite the importance of literacy in today's educational curriculum, learning to read is a challenge for many children with autism spectrum disorder (ASD). One of the foundational skills of early literacy learning is the ability to recognize sight words. This study used a single-subject, multiple-probe, across-participants design, to investigate the effects of a new software feature, dynamic text and speech output, on the acquisition of sight words by three pre-literate preschoolers with ASD during shared digital book reading experiences. All participants demonstrated successful acquisition of the target sight words with minimal exposure to the words. Limitations and future research directions are discussed, including the importance of investigating how the new software feature can be integrated into a more comprehensive literacy curriculum.


Degeneration and dysfunction of retinal neurons in acute ocular hypertensive rats: involvement of calpains.
Retinal ischemic diseases primarily lead to damage of the inner retinal neurons. Electrophysiological studies also suggest impairment of the inner retinal neurons. Our recent studies with acute ocular hypertensive rats confirmed damage predominantly in the inner retinal layer along with the ganglion cell layer, changes that are ameliorated by the calpain inhibitor SNJ-1945. However, we do not know which specific neuronal cells in the inner retinal layer are damaged by calpains. Thus, the purpose of the present study was to identify specific calpain-damaged neuronal cells in the inner retina from acute ocular hypertensive rats. Intraocular pressure was elevated to 110 mm Hg for 40 min. One hour after ocular hypertension (OH), SNJ-1945 was administrated as a single oral dose of 50 mg/kg. Retinal function was assessed by scotopic electroretinography (ERG). Histological degeneration was evaluated by hematoxylin and eosin, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end-labeling (TUNEL), and immunostaining in thin sections and flat mounts of the retina. Calpain activation was determined by proteolysis of the calpain substrate α-spectrin. OH caused calpain activation, increased TUNEL-positive staining, decreased thickness of the inner nuclear layer (INL), and decreased amplitudes of the ERG a- and b-waves and oscillatory potentials (OPs). SNJ-1945 significantly inhibited calpain activation and the decrease in ERG values. Interestingly, the changes in the b-wave and OPs amplitudes were significantly correlated to changes in the thickness of the INL. In the inner retinal layer, the numbers of rod bipolar, cone-ON bipolar, and amacrine cells were decreased after OH. SNJ-1945 suppressed the loss of cone-ON bipolar and amacrine cells, but did not inhibit the loss of rod bipolar cells. We also observed increased glial fibrillary acid protein-positive staining in the Müller cells after OH and the treatment with SNJ-1945. Calpains may contribute to ischemic retinal dysfunction by causing the loss of cone-ON bipolar and amacrine cells and causing the activation of Müller cells. Calpain inhibitor SNJ-1945 may be a candidate compound for treatment of retinal ischemic disease.


Misidentification of methicillin-resistant Staphylococcus aureus (MRSA) in hospitals in Tripoli, Libya.
Methicillin-resistant Staphylococcus aureus (MRSA) is a nosocomial (hospital-acquired) pathogen of exceptional concern. It is responsible for life-threatening infections in both the hospital and the community. To determine the frequency of MRSA misidentification in hospitals in Tripoli, Libya using current testing methods. One hundred and seventy S. aureus isolates previously identified as MRSA were obtained from three hospitals in Tripoli. All isolates were reidentified by culturing on mannitol salt agar, API 20 Staph System and retested for resistance to methicillin using the cefoxitin disk diffusion susceptibility test and PBP2a. D-tests and vancomycin E-tests (Van-E-tests) were also performed for vancomycin-resistant isolates. Of the 170 isolates examined, 86 (51%) were confirmed as MRSA (i.e. 49% were misidentified as MRSA). Fifteen (17%) of the confirmed MRSA strains exhibited inducible clindamycin resistance. Of the 86 confirmed MRSA isolates, 13 (15%) were resistant to mupirocin, 53 (62%) were resistant to ciprofloxacin, 41 (48%) were resistant to trimethoprim-sulfamethoxazole, and none were resistant to linezolid. Although disc-diffusion testing indicated that 23 (27%) of the isolates were resistant to vancomycin, none of the isolates were vancomycin-resistant by Van-E-test. Misidentification of nosocomial S. aureus as MRSA is a serious problem in Libyan hospitals. There is an urgent need for the proper training of microbiology laboratory technicians in standard antimicrobial susceptibility procedures and the implementation of quality control programs in microbiology laboratories of Libyan hospitals.


Cystic hydatidosis in slaughtered goats from various municipal abattoirs in Oman.
A passive surveillance study was conducted from April 2012 to April 2013 to determine the incidence, the affect of age, sex and cyst location, fertility, and viability on the occurrence of cystic hydatidosis in slaughtered goats from six municipal abattoirs around Oman. Morphologic characterization of suspected visceral organs has shown that Echinococcos granulosus (E. granulosus) metacestodes were present in 9.2 % of the total samples (682). Incidence (per 10,000) varied from 1.4 to 1.5. There was a significant difference between abattoirs, with Salalah having the highest infection (63.2 %), followed by Nizwa (27.2 %), Al Buraimi (10.8 %), Bousher (7.2 %), and the lowest in Sohar (1.6 %). Infection was significantly higher in indigenous goats (22.1 %) compared with imported breeds (2.5 %) from Somalia. The visceral organs were found harboring one or more hydatid cysts. Greatest cyst infection was recorded in lungs (53.4 %), followed by heart (50 %), liver (6.2 %), and multiple organs (14 %). A significant difference was observed between females (25 %) and male goats (6 %). Furthermore, the fertility of the hydatid cysts was found higher in heart and lungs (100 %, 51.6 %) and the viability rate of protoscolices was high (64 %). The findings of this study demonstrated that cystic hydatidosis is common and widely distributed in local goats, and they might play an important role in the life cycle and transmission of this zoonosis in Oman.


Specialist adult ADHD clinics in East Anglia: service evaluation and audit of NICE guideline compliance.
Aims and method To measure compliance with National Institute for Health and Care Excellence (NICE) recommendations in two adult attention-deficit hyperactivity disorder (ADHD) clinics and to guide further service development. We audited the case notes of 150 patients referred to adult ADHD clinics in East Anglia in 2010-2011 against NICE standards using an adapted version of the ADHD audit support tool. Results We found good compliance with NICE standards for diagnosis, assessment and pharmacological treatment of adult ADHD. There was a failure in smooth transitional arrangements from child and adolescent mental health to adult ADHD services. Comprehensive treatment programmes addressing psychological, behavioural, educational and occupational needs were not well developed. Deficiencies were observed in conducting recommended physical examinations. Substance use was prevalent in almost half of ADHD patients. Clinical implications Greater attention is needed in delivering better transitional arrangements and comprehensive treatment programmes for adult ADHD. More structured training with emphasis on ADHD-specific psychological interventions, physical examination and treatment of complex cases, especially with comorbid substance misuse, should be offered to clinicians.


Metformin suppresses adipogenesis through both AMP-activated protein kinase (AMPK)-dependent and AMPK-independent mechanisms.
People with Type 2 diabetes mellitus (T2DM) have reduced bone mineral density and an increased risk of fractures due to altered mesenchymal stem cell (MSC) differentiation in the bone marrow. This leads to a shift in the balance of differentiation away from bone formation (osteogenesis) in favour of fat cell development (adipogenesis). The commonly used anti-diabetic drug, metformin, activates the osteogenic transcription factor Runt-related transcription factor 2 (Runx2), which may suppress adipogenesis, leading to improved bone health. Here we investigate the involvement of the metabolic enzyme, AMP-activated protein kinase (AMPK), in these protective actions of metformin. The anti-adipogenic actions of metformin were observed in multipotent C3H10T1/2 MSCs, in which metformin exerted reciprocal control over the activities of Runx2 and the adipogenic transcription factor, PPARγ, leading to suppression of adipogenesis. These effects appeared to be independent of AMPK activation but rather through the suppression of the mTOR/p70S6K signalling pathway. Basal AMPK and mTOR/p70S6K activity did appear to be required for adipogenesis, as demonstrated by the use of the AMPK inhibitor, compound C. This observation was further supported by using AMPK knockout mouse embryo fibroblasts (MEFs) where adipogenesis, as assessed by reduced lipid accumulation and expression of the adipogeneic transcription factor, C/EBPβ, was found to display an absolute requirement for AMPK. Further activation of AMPK in wild type MEFS, with either metformin or the AMPK-specific activator, A769662, was also associated with suppression of adipogenesis. It appears, therefore, that basal AMPK activity is required for adipogenesis and that metformin can inhibit adipogenesis through AMPK-dependent or -independent mechanisms, depending on the cellular context.


One hundred years of the teaching of medicine in Singapore.
The Medical School was founded in 1905 in response to the need to train doctors for Singapore. In 2005, this School, now the Faculty of Medicine in the National University of Singapore, will celebrate her 100th year. This brief review attempts to chronicle the way in which the medical course has been designed over the years to address the principal aims of undergraduate medical education. From the inception of the Straits and Federated Malay States Government Medical School to the present, strong clinically based teaching was the key element, although there was some distinction between the foundational preclinical years and the clinical phases of training. Initially, this was necessitated by the fact that students were too poorly equipped in the sciences to have a more integrated curriculum. Over time, the structure of the course, learning objectives and teaching pedagogy and techniques became better developed. Of late, many of these elements have been revised and better articulated by the Faculty following a review. The method, frequency and emphasis of student evaluations have also changed as medical knowledge grows and the population becomes developed, with attendant disease pattern changes. The clinical relevance of the course and the need to train competent doctors remain the main mission of the Faculty of Medicine even in this age of molecular medicine and a technology-enhanced practice setting. Competency alone is, however, incomplete without good role models in the art of the practice of medicine. It is heartening to note that medical ethics and communications skills continue to be emphasised in the modern curriculum.


Making surgical wards safer for patients with diabetes: reducing hypoglycaemia and insulin errors.
High-quality perioperative diabetes care is essential to improve surgical outcomes for patients with diabetes. Inadequate perioperative diabetes care is associated with increased wound complications, higher mortality rates and increased length of hospital stay. Despite national guidelines, surgical wards remain a high-risk area for poor diabetes care. An initial baseline audit in 2014 of vascular patients with diabetes undergoing major lower limb amputation identified poor glycaemic control in 90% of patients, with high rates of hypoglycaemia and insulin management errors in 75%. Less than 15% of patients received specialist diabetes input and 20% required third-party assistance for hypoglycaemia. This quality improvement project aimed to reduce hypoglycaemia, insulin management errors and patient harm events by 50% in vascular surgery patients over a 3-year period. Key interventions over three successive Plan, Do, Study, Act cycles included educational and guideline initiatives (2015), establishing a diabetes in-reach service (2016) and implementing a whiteboard sugar cube alert system for poor glycaemic control (2017). The final introduction of the whiteboard sugar cube alert system delivered the greatest impact in reducing hypoglycaemia rates by more than 50%, insulin management errors by 70% and patient harm events by 75%.


Early signs of architectural and biomechanical failure in isolated myofibers and immortalized myoblasts from desmin-mutant knock-in mice.
In striated muscle, desmin intermediate filaments interlink the contractile myofibrillar apparatus with mitochondria, nuclei, and the sarcolemma. The desmin network's pivotal role in myocytes is evident since mutations in the human desmin gene cause severe myopathies and cardiomyopathies. Here, we investigated skeletal muscle pathology in myofibers and myofibrils isolated from young hetero- and homozygous R349P desmin knock-in mice, which carry the orthologue of the most frequent human desmin missense mutation R350P. We demonstrate that mutant desmin alters myofibrillar cytoarchitecture, markedly disrupts the lateral sarcomere lattice and distorts myofibrillar angular axial orientation. Biomechanical assessment revealed a high predisposition to stretch-induced damage in fiber bundles of R349P mice. Notably, Ca2+-sensitivity and passive myofibrillar tension were decreased in heterozygous fiber bundles, but increased in homozygous fiber bundles compared to wildtype mice. In a parallel approach, we generated and subsequently subjected immortalized heterozygous R349P desmin knock-in myoblasts to magnetic tweezer experiments that revealed a significantly increased sarcolemmal lateral stiffness. Our data suggest that mutated desmin already markedly impedes myocyte structure and function at pre-symptomatic stages of myofibrillar myopathies.


Genome-wide Mapping of Protein-DNA Interactions with ChEC-seq in Saccharomyces cerevisiae.
Genome-wide mapping of protein-DNA interactions is critical for understanding gene regulation, chromatin remodeling, and other chromatin-resident processes. Formaldehyde crosslinking followed by chromatin immunoprecipitation and high-throughput sequencing (X-ChIP-seq) has been used to gain many valuable insights into genome biology. However, X-ChIP-seq has notable limitations linked to crosslinking and sonication. Native ChIP avoids these drawbacks by omitting crosslinking, but often results in poor recovery of chromatin-bound proteins. In addition, all ChIP-based methods are subject to antibody quality considerations. Enzymatic methods for mapping protein-DNA interactions, which involve fusion of a protein of interest to a DNA-modifying enzyme, have also been used to map protein-DNA interactions. We recently combined one such method, chromatin endogenous cleavage (ChEC), with high-throughput sequencing as ChEC-seq. ChEC-seq relies on fusion of a chromatin-associated protein of interest to micrococcal nuclease (MNase) to generate targeted DNA cleavage in the presence of calcium in living cells. ChEC-seq is not based on immunoprecipitation and so circumvents potential concerns with crosslinking, sonication, chromatin solubilization, and antibody quality while providing high resolution mapping with minimal background signal. We envision that ChEC-seq will be a powerful counterpart to ChIP, providing an independent means by which to both validate ChIP-seq findings and discover new insights into genomic regulation.


Possible inhibitory effects of terbinafine on aripiprazole metabolism: Two case reports.
Aripiprazole, an atypical antipsychotic, is a metabolic substrate for cytochrome P450 (CYP)3A4 and 2D6. Terbinafine, an antifungal agent used for onychomycosis, is a CYP2D6 inhibitor and could theoretically reduce the metabolism of aripiprazole. However, there are no published reports describing this interaction. We present 2 female patients hospitalized in a psychiatric unit who were both taking aripiprazole 15 mg daily and terbinafine 250 mg daily prior to admission. The first patient was a 58-year-old female who was prescribed aripiprazole and terbinafine concomitantly for approximately 5 months prior to admission. A commercial pharmacogenetic testing platform classified this patient as a normal metabolizer for CYP3A4 and 2D6. The first patient's serum trough aripiprazole concentration at steady-state concentration (Css) was 207.5 ng/mL. The second patient was a 43-year-old female who was taking aripiprazole and terbinafine concomitantly for approximately 2 weeks prior to admission who had a Css aripiprazole concentration of 278.9 ng/mL. Aripiprazole has a wide therapeutic range (100 to 350 ng/mL) and a reference dose-related drug concentration of 11.7 (mean) ± 5.6 (SD) ng/mL/mg/d. Our patients had Css aripiprazole concentrations 18% and 59% higher than guideline-supported dose-related drug concentrations. Through the use of therapeutic drug monitoring, pharmacogenetic data, electronic pharmaceutical claims data, and the Drug Interaction Probability Scale, we suggest terbinafine possibly increases aripiprazole concentrations 18% to 59%. Further reports are needed to confirm these findings prior to using this information in clinical practice.


The effect of hydroalcoholic extract of P. crispum on sperm parameters, testis tissue and serum nitric oxide levels in mice.
Sperm dysfunction is one of the main causes of male infertility. Petroselinum crispum (P. crispum) is a member of umbelliferae family that contains different vitamins and minerals and has numerous therapeutic properties. The aim of this study was to evaluate P. crispum effect on sperm parameters, testis tissue and serum nitric oxid levels in mice. Hydroalcoholic extract of P. crispum was prepared and administered intraperitoneally (0,100, 150 and 200 mg/kg) to 40 mice, which were divided into four groups (n = 10), one control group and three experimental groups, for 14 consequent days. The sperm parameter such as motility, sperm count, morphology, and seminiferous tubules diameter, and weight of prostate and testis, and serum nitric oxide levels were analyzed. P. crispum administration (100, 150 and 200 mg/kg) significantly increased mean percentage of sperm motility, testis and prostate weight and serum nitric oxide compared to the control group (P < 0.05). However, no significant effect was reported for different doses of P. crispum extract on sperm parameters. Administrating hydroalcoholic extract of P. crispum has positive effects on some reproductive parameters.


Celastrol Alleviates Gamma Irradiation-Induced Damage by Modulating Diverse Inflammatory Mediators.
The present study aimed to explore the possible radioprotective effects of celastrol and relevant molecular mechanisms in an in vitro cell and in vivo mouse models exposed to gamma radiation. Human keratinocytes (HaCaT) and foreskin fibroblast (BJ) cells were exposed to gamma radiation of 20Gy, followed by treatment with celastrol for 24 h. Cell viability, reactive oxygen species (ROS), nitric oxide (NO) and glutathione (GSH) production, lipid peroxidation, DNA damage, inflammatory cytokine levels, and NF-κB pathway activation were examined. The survival rate, levels of interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α) in blood, and p65 and phospho-p65 expression were also evaluated in mice after exposure to gamma radiation and celastrol treatment. The gamma irradiation of HaCaT cells induced decreased cell viability, but treatment with celastrol significantly blocked this cytotoxicity. Gamma irradiation also increased free radical production (e.g., ROS and NO), decreased the level of GSH, and enhanced oxidative DNA damage and lipid peroxidation in cells, which were effectively reversed by celastrol treatment. Moreover, inflammatory responses induced by gamma irradiation, as demonstrated by increased levels of IL-6, TNF-α, and IL-1β, were also blocked by celastrol. The increased activity of NF-κB DNA binding following gamma radiation was significantly attenuated after celastrol treatment. In the irradiated mice, treatment with celastrol significantly improved overall survival rate, reduced the excessive inflammatory responses, and decreased NF-κB activity. As a NF-κB pathway blocker and antioxidant, celastrol may represent a promising pharmacological agent with protective effects against gamma irradiation-induced injury.


Self-perception and experiential schemata in the addicted brain.
This study investigated neurophysiological differences between recovering substance abusers (RSA) and controls while electroencephalogram (EEG) was continuously recorded during completion of a new assessment instrument. The participants consisted of 56 total subjects; 28 RSA and 28 non-clinical controls (C). The participants completed the self-perception and experiential schemata assessment (SPESA) and source localization was compared utilizing standardized low-resolution electromagnetic tomography (sLORETA). The data show significant differences between groups during both the assessment condition and baselines. A pattern of alpha activity as estimated by sLORETA was shown in the right amygdala, uncus, hippocampus, BA37, insular cortex and orbitofrontal regions during the SPESA condition. This activity possibly reflects a circuit related to negative perceptions of self formed in specific neural pathways. These pathways may be responsive to the alpha activity induced by many substances by bringing the brain into synchrony if only for a short time. In effect this may represent the euphoria described by substance abusers.


Mice null for NEDD9 (HEF1α) display extensive hippocampal dendritic spine loss and cognitive impairment.
NEDD9 (neural precursor cell expressed, developmentally down-regulated 9) is a member of the CAS (Crk-associated substrate) family of scaffolding proteins that regulate cell adhesion and migration. A Nedd9 knock-out/lacZ knock-in mouse (Nedd9(-/)(-)) was developed in order to study Nedd9 expression and function in the nervous system. Herein we show that NEDD9 is expressed in the adult brain and is prominently expressed in the hippocampus. Behavioral testing uncovered functional deficits in Nedd9(-)(/)(-) mice. In the Morris water maze test, Nedd9(-)(/)(-) mice showed deficits in both the ability to learn the task as well as in their ability to recall the platform location. There was no change in the gross morphology of the hippocampus, and stereological analysis of BrdU-labeled newly formed hippocampal cells suggested that this defect is not secondary to altered neurogenesis. However, analysis of the hippocampus revealed extensive loss of dendritic spine density in both the dentate gyrus (DG) and CA1 regions. Spine loss occurred equally across all branch orders and regions of the dendrite. Analysis of spine density in Nedd9(-)(/)(-) mice at 1.5, 6 and 10 months revealed an age-dependent spine loss. This work shows that NEDD9 is required for the maintenance of dendritic spines in the hippocampus, and suggests it could play a role in learning and memory.


Anthropometric measures and physical activity and the risk of lung cancer in never-smokers: a prospective cohort study.
Worldwide, lung cancer in never-smokers is ranked the seventh most common cause of cancer death; however, the etiology of lung cancer in never-smokers is unclear. We investigated associations for body mass index (BMI) at various ages, waist circumference, hip circumference, and physical activity with lung cancer in 158,415 never-smokers of the NIH-AARP Diet and Health Study. Multivariable hazard ratios (HR) and 95% confidence intervals (CI) were estimated from Cox proportional hazards models. Over 11 years of follow-up, 532 lung cancer cases occurred. The risk estimate for obese (BMI ≥ 30 kg/m(2)) participants at baseline was 1.21 (95%CI = 0.95-1.53) relative to those with a normal BMI between 18.5 ≤ BMI<25.0. Overweight (25.0 ≤ BMI<30.0) at age 18 (HR(overweight-vs-normal) = 1.51;95%CI = 1.01-2.26) and time spent sitting (HR(≥ 3 hrs-vs-<3 hrs) = 1.32;95%CI = 1.00-1.73) was each associated with lung cancer after adjustment for baseline BMI, as was waist (HR(Q4-vs-Q1) = 1.75;95%CI = 1.09-2.79) and hip circumference (HRQ4-vs-Q1 = 0.62;95%CI = 0.39-0.99), after mutual adjustment for each other and baseline BMI. No associations were observed for vigorous activity or television watching. In summary, using a large prospective cohort study, we found no evidence that BMI at baseline or middle age was associated with decreased lung cancer risk in never smokers. If anything, we observed some evidence for positive associations with a larger BMI or waist circumference.


Synaptic plasticity and dysconnection in schizophrenia.
Current pathophysiological theories of schizophrenia highlight the role of altered brain connectivity. This dysconnectivity could manifest 1) anatomically, through structural changes of association fibers at the cellular level, and/or 2) functionally, through aberrant control of synaptic plasticity at the synaptic level. In this article, we review the evidence for these theories, focusing on the modulation of synaptic plasticity. In particular, we discuss how dysconnectivity, observed between brain regions in schizophrenic patients, could result from abnormal modulation of N-methyl-D-aspartate (NMDA)-dependent plasticity by other neurotransmitter systems. We focus on the implication of the dysconnection hypothesis for functional imaging at the systems level. In particular, we review recent advances in measuring plasticity in the human brain using functional magnetic resonance imaging (fMRI) and electroencephalography (EEG) that can be used to address dysconnectivity in schizophrenia. Promising experimental paradigms include perceptual and reinforcement learning. We describe how theoretical and causal models of brain responses might contribute to a mechanistic understanding of synaptic plasticity in schizophrenia.