Connexin 43 Inhibition Sensitizes Chemoresistant Glioblastoma Cells to Temozolomide.
Resistance of glioblastoma (GBM) to the front-line chemotherapeutic agent temozolomide (TMZ) continues to challenge GBM treatment efforts. The repair of TMZ-induced DNA damage by O-6-methylguanine-DNA methyltransferase (MGMT) confers one mechanism of TMZ resistance. Paradoxically, MGMT-deficient GBM patients survive longer despite still developing resistance to TMZ. Recent studies indicate that the gap junction protein connexin 43 (Cx43) renders GBM cells resistant to TMZ through its carboxyl terminus (CT). In this study, we report insights into how Cx43 promotes TMZ resistance. Cx43 levels were inversely correlated with TMZ sensitivity of GBM cells, including GBM stem cells. Moreover, Cx43 levels inversely correlated with patient survival, including as observed in MGMT-deficient GBM patients. Addition of the C-terminal peptide mimetic αCT1, a selective inhibitor of Cx43 channels, sensitized human MGMT-deficient and TMZ-resistant GBM cells to TMZ treatment. Moreover, combining αCT1 with TMZ-blocked AKT/mTOR signaling, induced autophagy and apoptosis in TMZ-resistant GBM cells. Our findings suggest that Cx43 may offer a biomarker to predict the survival of patients with MGMT-independent TMZ resistance and that combining a Cx43 inhibitor with TMZ could enhance therapeutic responses in GBM, and perhaps other TMZ-resistant cancers.


The Brain Microenvironment Induces DNMT1 Suppression and Indolence of Metastatic Cancer Cells.
Brain metastasis is an ineffective process, and many cancer cells enter into an indolent state following extravasation in the brain. Single cell RNA sequencing of melanoma brain metastases reveals that non-proliferating brain metastatic melanoma cells exhibit a pattern of gene expression associated with inhibition of DNA methyltransferase 1 (DNMT1). The brain microenvironment, specifically the combination of reactive astrocytes and mechanically soft surroundings, suppressed DNMT1 expression in various cancer types and caused cell cycle delay. Somewhat unexpectedly, we find that DNMT1 suppression not only induces cell cycle delay but also activates pro-survival signals in brain metastatic cancer cells, including L1CAM and CRYAB. Our results demonstrate that transcriptional changes triggered by DNMT1 suppression is a key step for cancer cells to survive in the brain microenvironment and that they also restrict cancer cell proliferation. The dual consequences of DNMT1 suppression can explain the persistence of indolent cancer cells in the brain microenvironment.


Acinar cell-specific knockout of the PTHrP gene decreases the proinflammatory and profibrotic responses in pancreatitis.
Pancreatitis is a necroinflammatory disease with acute and chronic manifestations. Accumulated damage incurred during repeated bouts of acute pancreatitis (AP) can lead to chronic pancreatitis (CP). Pancreatic parathyroid hormone-related protein (PTHrP) levels are elevated in a mouse model of cerulein-induced AP. Here, we show elevated PTHrP levels in mouse models of pancreatitis induced by chronic cerulein administration and pancreatic duct ligation. Because acinar cells play a major role in the pathophysiology of pancreatitis, mice with acinar cell-specific targeted disruption of the Pthrp gene (PTHrP(Δacinar)) were generated to assess the role of acinar cell-secreted PTHrP in pancreatitis. These mice were generated using Cre-LoxP technology and the acinar cell-specific elastase promoter. PTHrP(Δacinar) exerted protective effects in cerulein and pancreatic duct ligation models, evident as decreased edema, histological damage, amylase secretion, pancreatic stellate cell (PSC) activation, and extracellular matrix deposition. Treating acinar cells in vitro with cerulein increased IL-6 expression and NF-κB activity; these effects were attenuated in PTHrP(Δacinar) cells, as were the cerulein- and carbachol-induced elevations in amylase secretion. The cerulein-induced upregulation of procollagen I expression was lost in PSCs from PTHrP(Δacinar) mice. PTHrP immunostaining was elevated in human CP sections. The cerulein-induced upregulation of IL-6 and ICAM-1 (human acinar cells) and procollagen I (human PSCs) was suppressed by pretreatment with the PTH1R antagonist, PTHrP (7-34). These findings establish PTHrP as a novel mediator of inflammation and fibrosis associated with CP. Acinar cell-secreted PTHrP modulates acinar cell function via its effects on proinflammatory cytokine release and functions via a paracrine pathway to activate PSCs.


Development of gut colonisation in pre-term neonates.
Twenty-eight pre-term babies of low birth weight were monitored for developing microflora in throat, stomach and faeces during the first 3 weeks of life. The flora at all levels of the gastrointestinal tract differed from that of healthy breast-fed and artificially fed full-term babies. Colonisation of throat and stomach was delayed beyond 4 days of life in 87% and 60% of babies respectively. Only 10% of babies had "normal" oral flora throughout the period of study. Flora of the stomach was sparse, and resembled faecal flora. Faecal flora was established more rapidly than throat or stomach flora, and 70% of babies were colonised during the first 4 days of life. Initially Bacteroides spp. were predominant (57% babies), but Escherichia coli and other aerobic gram-negative bacilli gradually increased in frequency. Colonisation by gram-positive bacteria was slow. Clostridium spp. were present in only 10% of babies during the first 4 days of life. Most strains were transient. Colonisation with C. butyricum (30%), C. perfringens (35%) and C. difficile (25%) was maximum after the first 2 weeks of life. Lactic-acid-producing bacteria usually appeared late in the third week of life. Parenteral feeding immediately after birth was associated with delayed colonisation by a restricted number of species. Parenteral antibiotics (penicillin or gentamicin or both) restricted colonisation with normal oral flora, the lactic-acid-producing bacteria and penicillin-sensitive clostridia, but had little effect on E. coli even when the colonising strain was sensitive to the aminoglycoside in the regimen. Systemic spread of bacteria via the blood stream was not detected in any babies. The pattern of colonisation of the enteric tract in pre-term infants in the special-care nursery studied, differs from that of healthy full-term babies; this merits consideration when the results of bacteriological tests of this vulnerable group of infants are being interpreted.


A Model of Repetitive-DNA-Organized Chromatin Network of Interphase Chromosomes.
During interphase, chromosomes are relatively de-condensed in the nuclear space. Interphase chromosomes are known to occupy nuclear space in a non-random manner (chromosome territory); however, their internal structures are poorly defined. In particular, little is understood about the molecular mechanisms that govern the internal organization of interphase chromosomes. The author recently proposed that pairing (or interaction) of repetitive DNA-containing chromatin regions is a critical driving force that specifies the higher-order organization of eukaryotic chromosomes. Guided by this theoretical framework and published experimental data on the structure of interphase chromosomes and the spatial distribution of repetitive DNA in interphase nuclei, I postulate here a molecular structure of chromatin organization in interphase chromosomes. According to this model, an interphase chromosome is a chromatin mesh (or lattice) that is formed by repeat pairing (RP). The mesh consists of two types of structural components: chromosome nodes and loose chromatin fibers. Chromosome nodes are DNA repeat assemblies (RAs) that are formed via RP, while loose fibers include chromatin loops that radiate from the nodes. Different loops crosslink by RPs and form a large integrated chromatin network. I suggest that the organization of the chromatin network of a given interphase chromosome is intrinsically specified by the distribution of repetitive DNA elements on the linear chromatin. The stability of the organization is governed by the collection of RA-formed nodes, and the dynamics of the organization is driven by the assembling and disassembling of the nodes.


Optical image processing for synthetic-aperture imaging ladar based on two-dimensional Fourier transform.
A two-dimensional (2D) Fourier transform algorithm for the image reconstruction of synthetic-aperture imaging ladar (SAIL) collected data is suggested. This algorithm consists of quadratic phase compensation in azimuth direction and 2D fast Fourier transform. Based on this algorithm and the parallel 2D Fourier transform capability of spherical lens, an optical principle scheme that processes the SAIL data is proposed. The basic principle, design equations, and necessary analysis are presented. To verify this principle scheme, an experimental optical SAIL processor setup is constructed. The imaging results of SAIL data obtained by our SAIL demonstrator are presented. The optical processor is compact, lightweight, and consumes low power. This optical processor can also provide inherent parallel and speed-of-light computing capability, and thus has potential applications in on-board and satellite-borne SAIL systems.


Does the duration of abdominal pain prior to admission influence the severity of acute pancreatitis?
In a prior report involving patients with hemoconcentration at admission, those with necrotizing pancreatitis presented significantly earlier than those with interstitial disease suggesting that duration of abdominal pain prior to presentation may have prognostic significance in acute pancreatitis. The aim of the present study was to determine whether the duration of abdominal pain prior to admission influences the severity of acute pancreatitis. During a five-year period, all patients presenting directly to our hospital with their first episode of acute pancreatitis were enrolled in a cohort study. We analyzed data obtained from records of all such patients and performed a separate analysis on those with hemoconcentration (hematocrit equal to, or greater than, 44%) at presentation to determine whether duration of abdominal pain prior to presentation was associated with severity of acute pancreatitis. Duration of abdominal pain was categorized as persisting for either less than 12 h or 12 h or more prior to arrival. Prognostic markers of severity included admission hematocrit and blood urea nitrogen (BUN), as well as the development of systemic inflammatory response syndrome (SIRS) during the initial 24 h of hospitalization. Outcome measures included pancreatic necrosis based on contrast-enhanced CT scanning, need for intensive care, length of hospitalization, and death. Radiologic severity of peripancreatic inflammatory changes was assessed within 48 h of admission in accordance with the Balthazar-Ranson scoring system (A-E). Among a total of 318 patients, there were 62 (19.5%) with hemoconcentration at admission. Among the 318 patients, there was no significant difference in the prevalence of pancreatic necrosis when comparing the less than 12 h group to the 12 h or more group. Among the 62 patients with hemoconcentration, those admitted within 12 h compared to those admitted 12 h or more following the onset of abdominal pain had an increased radiologic severity of acute pancreatitis (Balthazar-Ranson grade D or E: 83.3% vs. 40.0%; P=0.006) and an increased prevalence of pancreatic necrosis (21.1% vs. 2.3%; P=0.028). Duration of abdominal pain prior to admission impacts the severity of acute pancreatitis only among patients with hemoconcentration at presentation.


Elucidating mechanisms of osteogenesis in human adipose-derived stromal cells via microarray analysis.
The osteogenic potential of human adipose-derived stromal cells (hASCs), the ease of cell procurement, and the shortcomings of conventional skeletal reconstruction call for further analysis of the molecular mechanisms governing hASC osteogenic differentiation. We have examined the expression profile of the human transcriptome during osteogenic differentiation of ASCs using microarray. Subsequently, we analyzed those genes related to osteogenesis that have not been previously studied about hASCs. We have preliminarily assessed the role of IGFBP3, TGF-B3, TNC, CTGF, DKK-1, and PDGFRB in hASC osteogenic differentiation. We compared the expression profile of undifferentiated hASCs to that of hASCs treated with osteogenic differentiation medium for 1, 3, or 7 days using the Human Exonic Evidence-Based Oligonucleotide chip. Genes significantly overexpress or underexpressed were validated with quantitative reverse transcription-polymerase chain reaction. The osteogenic capability of ASCs was verified by Alizarin Red staining. IGFBP3, TGF-B3, TNC, CTGF, and PDGFRB were all upregulated in early osteogenesis, and TGF-B3, TNC, and PDGFRB were upregulated in late osteogenesis by microarray and quantitative reverse transcription analysis. In contrast, DKK-1 was downregulated in early and late osteogenesis. Alizarin Red staining showed a significant increase in mineralization in hASCs, even after 1 day in osteogenic differentiation medium. Factors that commit hASCs to an osteogenic pathway remain largely unknown. We have described 6 genes that play key roles in hASC osteogenic differentiation. We plan to further exploit these data via in vitro treatment of hASCs with these soluble cytokines and in vivo translation using a nude mouse calvarial defect model.


Impaired cerebral mitochondrial function after traumatic brain injury in humans.
Oxygen supply to the brain is often insufficient after traumatic brain injury (TBI), and this results in decreased energy production (adenosine triphosphate [ATP]) with consequent neuronal cell death. It is obviously important to restore oxygen delivery after TBI; however, increasing oxygen delivery alone may not improve ATP production if the patient's mitochondria (the source of ATP) are impaired. Traumatic brain injury has been shown to impair mitochondrial function in animals; however, no human studies have been previously reported. Using tissue fractionation procedures, living mitochondria derived from therapeutically removed brain tissue were analyzed in 16 patients with head injury (Glasgow Coma Scale Scores 3-14) and two patients without head injury. Results revealed that in head-injured patients mitochondrial function was impaired, with subsequent decreased ATP production. Decreased oxygen metabolism due to mitochondrial dysfunction must be taken into account when clinically defining ischemia and interpreting oxygen measurements such as jugular venous oxygen saturation, arteriovenous difference in oxygen content, direct tissue oxygen tension, and cerebral blood oxygen content determined using near-infrared spectroscopy. Restoring mitochondrial function might be as important as maintaining oxygen delivery.


VALERIE: Visual-based inspection of alternative splicing events at single-cell resolution.
We present VALERIE (Visualising alternative splicing events from single-cell ribonucleic acid-sequencing experiments), an R package for visualising alternative splicing events at single-cell resolution. To explore any given specified genomic region, corresponding to an alternative splicing event, VALERIE generates an ensemble of informative plots to visualise cell-to-cell heterogeneity of alternative splicing profiles across single cells and performs statistical tests to compare percent spliced-in (PSI) values across the user-defined groups of cells. Among the features available, VALERIE displays PSI values, in lieu of read coverage, which is more suitable for representing alternative splicing profiles for a large number of samples typically generated by single-cell RNA-sequencing experiments. VALERIE is available on the Comprehensive R Archive Network (CRAN): https://cran.r-project.org/web/packages/VALERIE/index.html.


Impact of different diagnostic technologies for MRSA admission screening in hospitals - a decision tree analysis.
Hospital infections with multiresistant bacteria, e.g., Methicillin-resistant Staphylococcus aureus (MRSA), cause heavy financial burden worldwide. Rapid and precise identification of MRSA carriage in combination with targeted hygienic management are proven to be effective but incur relevant extra costs. Therefore, health care providers have to decide which MRSA screening strategy and which diagnostic technology should be applied according to economic criteria. The aim of this study was to determine which MRSA admission screening and infection control management strategy causes the lowest expected cost for a hospital. Focus was set on the Point-of-Care Testing (PoC). A decision tree analytic cost model was developed, primarily based on data from peer-reviewed literature. In addition, univariate sensitivity analyses of the different input parameters were conducted to study the robustness of the results. In the basic analysis, risk-based PoC screening showed the highest mean cost savings with 14.98 € per admission in comparison to no screening. Rapid universal screening methods became favorable at high MRSA prevalence, while in situations with low MRSA transmission rates omission of screening may be favorable. Early detection of MRSA by rapid PoC or PCR technologies and consistent implementation of appropriate hygienic measures lead to high economic efficiency of MRSA management. Whether general or targeted screening is more efficient depends mainly on epidemiological and infrastructural parameters.


Quantitative real-time PCR using TaqMan and SYBR Green for Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, tetQ gene and total bacteria.
Accurate quantification of bacterial species in dental plaque is needed for microbiological diagnosis of periodontal diseases. The present study was designed to assess the sensitivity, specificity and quantitativity of the real-time PCR using the GeneAmp Sequence Detection System with two fluorescence chemistries. TaqMan probe with reporter and quencher dye, and SYBR Green dye were used for sources of the fluorescence. Primers and probes were designed for Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia and total bacteria based on the nucleotide sequences of the respective 16S ribosomal RNA genes. Since spread of antibiotic resistance genes is one of the crucial problems in periodontal therapy, quantitative detection of tetQ gene, which confers resistance to tetracycline, was included in the examination. The detection of P. gingivalis, P. intermedia and A. actinomycetemcomitans was linear over a range of 10-10(7) cells (10-10(7) copies for tetQ gene), while the quantitative range for total bacteria was 10(2)-10(7) cells. Species-specific amplifications were observed for the three periodontal bacteria, and there was no significant difference between the TaqMan and SYBR Green chemistry in their specificity, quantitativity and sensitivity. The SYBR Green assay, which was simpler than TaqMan assay in its manipulations, was applied to the clinical plaque samples. The plaque samples were obtained from eight patients (eight periodontal pockets) before and 1 week after the local drug delivery of minocycline. Although the number of P. gingivalis, P. intermedia and A. actinomycetemcomitans markedly decreased after the antibiotic therapy in most cases, higher copy numbers of the tetQ gene were detectable. The real-time PCR demonstrated sufficient sensitivity, specificity and quantitativity to be a powerful tool for microbiological examination in periodontal disease, and the quantitative monitoring of antibiotic resistance gene accompanied with the antibiotic therapy should be included in the examination.


Interferon (IFN) beta acts downstream of IFN-gamma-induced class II transactivator messenger RNA accumulation to block major histocompatibility complex class II gene expression and requires the 48-kD DNA-binding protein, ISGF3-gamma.
Interferon (IFN) gamma, a cardinal proinflammatory cytokine, induces expression of the gene products of the class II locus of the major histocompatibility complex (MHC), whereas IFN-alpha or -beta suppresses MHC class II expression. The mechanism of IFN-beta-mediated MHC class II inhibition has been unclear. Recently, a novel factor termed class II transactivator (CIITA) has been identified as essential for IFN-gamma-induced MHC class II transcription. We studied the status of IFN-gamma-induced CIITA messenger RNA (mRNA) accumulation and CIITA-driven transactivation in IFN-beta-treated cells and used cell lines that had defined defects in the type I IFN response pathway to address the roles of IFN signaling components in the inhibition of MHC class II induction. IFN-beta treatment did not suppress IFN-gamma-induced accumulation of CIITA mRNA. After cells were stably transfected with CIITA, endogenous MHC class II genes were constitutively expressed, and MHC class II promoters, delivered by transfection, were actively transcribed in CIITA-expressing cells. Expression of these promoters was significantly impaired by pretreatment with IFN-beta. These results suggest that IFN-beta acts downstream of CIITA mRNA accumulation, and acts in part by reducing the functional competence of CIITA for transactivating MHC class II promoters. IFN stimulated gene factor 3 (ISGF3) gamma was essential for IFN-beta to mediate inhibition of MHC class II induction, regardless of whether MHC class II transcription was stimulated by IFN-gamma or directly by CIITA expression. Results of these experiments suggest that inhibition of MHC class II in IFN-beta-treated cells requires expression of gene(s) directed by the ISGF3-IFN-stimulated response element pathway, and that these gene product(s) may act by blocking CIITA-driven transcription of MHC class II promoters.


Nonself vegetative fusion and genetic exchange in the arbuscular mycorrhizal fungus Glomus intraradices.
Arbuscular mycorrhizal fungi (AMF) form symbioses with the majority of plants and form extensive underground hyphal networks simultaneously connecting the roots of different plant species. No empirical evidence exists for either anastomosis between genetically different AMF or genetic exchange.Five isolates of one population of Glomus intraradices were used to study anastomosis between hyphae of germinating spores. We show that genetically distinct AMF, from the same field, anastomose, resulting in viable cytoplasmic connections through which genetic exchange could potentially occur.Pairs of genetically different isolates were then co-cultured in an in vitro system.Freshly produced spores were individually germinated to establish new cultures.Using several molecular tools, we show that genetic exchange occurred between genetically different AMF. Specific genetic markers from each parent were transmitted to the progeny. The progeny were viable, forming symbioses with plant roots. The phenotypes of some of the progeny were significantly different from either parent.Our results indicate that considerable promiscuity could occur in these fungi because nine out of 10 combinations of different isolates anastomosed. The ability to perform genetic crosses between AMF experimentally lays a foundation for understanding the genetics and evolutionary biology of these important plants symbionts.


Impaired somatic growth and delayed lung development in infants with congenital diaphragmatic hernia--evidence from a 10-year, single center prospective follow-up study.
In infants with congenital diaphragmatic hernia (CDH), somatic growth and pulmonary development are key issues beyond the time of intensive care treatment. The aim of the study was to investigate the somatic growth and pulmonary function after discharge and to compare CDH patients with a group of matched controls. Anthropometric measurements and lung function tests were performed in 26 infants after surgical repair of CDH and 26 non-CDH intensive care patients, matched for gestational age and birth weight. Spontaneously breathing infants were tested at a mean of 44 weeks postconceptional age (range, 36-58 weeks). Body weight, body length, respiratory rate (RR), tidal volume (V(T)), functional residual capacity by body plethysmography (FRC(pleth)), respiratory compliance (C(rs)), and respiratory resistance (R(rs)) were measured. The mean (SD) weight gain per week in the CDH infants was significantly lower compared to non-CDH infants (89 [39] g vs 141 [49] g; P = .002). The breathing pattern between both groups differed considerably. In CDH infants, V(T) was significantly lower (P < .001) and RR significantly higher (P = .005). The respiratory compliance was also significantly (P < .001) reduced, whereas R(rs) did not differ significantly. No statistically significant differences were seen in FRC(pleth) related to the body weight between CDH and non-CDH infants (20.3 [4.4] mL/kg vs 21.5 [4.9] mL/kg). Despite apparently well-inflated lungs after surgery, evidence of early and significantly reduced weight gain and impaired lung function in CHD patients should prompt careful dietary monitoring and regular lung function testing.


Effect of curcumin on the proliferation, apoptosis, migration, and invasion of human melanoma A375 cells.
Malignant melanoma is a melanocytic tumor with a high potential of invasion and metastasis. Curcumin is extracted from Curcuma longa L.; curcumin has anti-tumor efficacy in multiple systemic malignancies. Here, we investigated the effect of curcumin on A375 human melanoma cells. A375 cells were cultivated, passaged, and treated with different concentrations of curcumin. We observed the cellular morphology and determined the migration, invasion, proliferation, and apoptosis of A375 cells in vitro. Our results showed that curcumin induced a significant change in the morphology of A375 cells. Compared to the control group, the groups treated with curcumin showed significantly wider scratches, and the number of A375 cells significantly decreased in the 12.5, 25, and 50 mM curcumin groups (P < 0.05 or < 0.01). The rates of proliferation inhibition in the 5 curcumin groups were 19.38 ± 3.57%, 35.56 ± 4.37%, 63.98 ± 5.95%, 86.38 ± 3.91%, and 95.56 ± 3.15%. The half-maximal inhibitory concentration of curcumin at 48 h was 10.05 mM. The rates of apoptosis in 6.25 and 12.5 mM curcumin groups were significantly higher (P < 0.05), phosphorylation levels of JAK-2 and STAT-3 in 10 and 20 mM curcumin groups were significantly lower (P < 0.05), and Bcl-2 protein expression in 1, 2.5, 5, 10, and 20 curcumin groups was significantly lower (P < 0.05) than that in the control group. In conclusion, curcumin has antiproliferative and proapoptotic activities on A375 cells, the mechanism of which may be related to the inhibition of JAK-2/STAT-3 signaling pathway.


The NASSS framework for ex post theorisation of technology-supported change in healthcare: worked example of the TORPEDO programme.
Evaluation of health technology programmes should be theoretically informed, interdisciplinary, and generate in-depth explanations. The NASSS (non-adoption, abandonment, scale-up, spread, sustainability) framework was developed to study unfolding technology programmes in real time-and in particular to identify and manage their emergent uncertainties and interdependencies. In this paper, we offer a worked example of how NASSS can also inform ex post (i.e. retrospective) evaluation. We studied the TORPEDO (Treatment of Cardiovascular Risk in Primary Care using Electronic Decision Support) research programme, a multi-faceted computerised quality improvement intervention for cardiovascular disease prevention in Australian general practice. The technology (HealthTracker) had shown promise in a cluster randomised controlled trial (RCT), but its uptake and sustainability in a real-world implementation phase was patchy. To explain this variation, we used NASSS to undertake secondary analysis of the multi-modal TORPEDO dataset (results and process evaluation of the RCT, survey responses, in-depth professional interviews, videotaped consultations) as well as a sample of new, in-depth narrative interviews with TORPEDO researchers. Ex post analysis revealed multiple areas of complexity whose influence and interdependencies helped explain the wide variation in uptake and sustained use of the HealthTracker technology: the nature of cardiovascular risk in different populations, the material properties and functionality of the technology, how value (financial and non-financial) was distributed across stakeholders in the system, clinicians' experiences and concerns, organisational preconditions and challenges, extra-organisational influences (e.g. policy incentives), and how interactions between all these influences unfolded over time. The NASSS framework can be applied retrospectively to generate a rich, contextualised narrative of technology-supported change efforts and the numerous interacting influences that help explain its successes, failures, and unexpected events. A NASSS-informed ex post analysis can supplement earlier, contemporaneous evaluations to uncover factors that were not apparent or predictable at the time but dynamic and emergent.


Chorioamnionitis and Five-Year Neurodevelopmental Outcome in Preterm Infants.
Chorioamnionitis, a risk factor for preterm delivery, has been suggested to be associated with suboptimal neurological development in premature infants. To evaluate the association between chorioamnionitis and neurodevelopment in preterm infants at 5 years of age. Methods Very low birth weight and very low gestational age infants (n = 197) were recruited. Placental samples (n = 117) were evaluated for histological chorioamnionitis. Fetal histological chorioamnionitis was analyzed as a subgroup. The diagnosis of clinical chorioamnionitis was derived from medical records. Neurodevelopmental impairments were evaluated at 2 years of age, and cognitive development (n = 188) and neuropsychological performance (n = 193) were evaluated at 5 years of age. There were no associations between histological or clinical chorioamnionitis and neurodevelopmental impairments at 2 years of age. Clinical chorioamnionitis and fetal histological chorioamnionitis were not associated with cognitive development or neuropsychological performance, but histological chorioamnionitis was associated with poorer cognitive outcome (regression coefficient = -7.22, 95% CI: -14.31 to -0.13) and weaker memory and learning functions (regression coefficient = -1.29, 95% CI: -2.40 to -0.18) at 5 years of age. Our study findings do not support clinical chorioamnionitis having a major independent role in the pathogenesis of neurodevelopmental problems in very preterm infants. Histological chorioamnionitis was associated with slightly less optimal performance at 5 years of age, but further studies are needed to verify the clinical significance of these findings.


Molecular characterisation of chondrocytes in autologous chondrocyte implantation.
Autologous chondrocyte implantation (ACI) relies on the use of cultured cells. However, the biosynthetic profile of cultured chondrocytes is shown to be altered during in vitro expansion. The purpose of this study therefore, was to examine the cellular phenotype of chondrocytes cultured for ACI and to determine the apoptotic index of cells implanted into patients. Using electron microscopy, immunohistochemistry, RT-PCR and flow cytometry analyses, we have investigated protein and gene expression of several chondrocyte-specific, or associated markers in cultured cells used for implantation in patients. They included S-100, type I and II collagen, aggrecan, transforming growth factor beta, glucocorticoid receptor alpha and beta and vitamin D3 receptor. We have also examined the apoptotic index of chondrocytes. Our results demonstrated that cultured cells for ACI display the characteristics of chondrocytes. These cells are round in shape, contain numerous small surface processes of cytoplasmic membrane and have an accumulation of glycogen within the cytoplasm. They express S-100, aggrecan TGF-beta, glucocorticoid receptor alpha and vitamin D3 receptor as evidenced by either immunohistochemistry or RT-PCR, however, there is variation in the expression of type I, type II collagen glucocorticoid receptor beta between cases. Chondrocyte used for implantation has relatively low level of apoptosis (<11%). In conclusion, although there was variation in the level of expression of these genetic markers, our data indicate that cultured cells used for ACI were of chondrocytic lineage cells and have low level of apoptotic cells.


Epileptic psychoses - evaluation of clinical aspects.
Epileptic psychoses as the most complex psychopathological phenomena represent unexplored states for prognosis. Clinical trials, conveyed in order to found risk factors still are not consistent in their conclusions. By this research results of all biological, clinical, psychological and social, as well as demographic factors will contribute to opportunity to find variables which could finally prevent these conditions. In this study we tried to evaluate clinical variables which could foresee manifestation of interictal and postictal psychosis. This research study is epidemiological, clinical, retrospective and analytical. In total 567 patients were included in this study, which belonged to the diagnostic criteria F 0.6 according to ICD-10 classification, among which 14 patients with the diagnosed epileptic psychosis (06,8) were followed. All patients were hospitalized and evaluated at the Psychiatric Clinic in University of Sarajevo Clinics Centre, during time period between 01.01.2000 - 31.08.2006. Within baseline all relevant clinical and demographic variables were evaluated. Among patients most dominant form of behavior was expansive, with emphasized paranoid ideations and perception of auditory hallucinations. A correlation between intensive psychological trauma as an exacerbation factor and prolonged illness is determined, as well as between expression of psychotic symptoms and forced normalization by antiepileptic medications. Also is proven that among postictal psychosis more dominant are suicide attempts and aggressive behavior.


Distinction between Externally vs. Internally Guided Decision-Making: Operational Differences, Meta-Analytical Comparisons and Their Theoretical Implications.
Most experimental studies of decision-making have specifically examined situations in which a single less-predictable correct answer exists (externally guided decision-making under uncertainty). Along with such externally guided decision-making, there are instances of decision-making in which no correct answer based on external circumstances is available for the subject (internally guided decision-making). Such decisions are usually made in the context of moral decision-making as well as in preference judgment, where the answer depends on the subject's own, i.e., internal, preferences rather than on external, i.e., circumstantial, criteria. The neuronal and psychological mechanisms that allow guidance of decisions based on more internally oriented criteria in the absence of external ones remain unclear. This study was undertaken to compare decision-making of these two kinds empirically and theoretically. First, we reviewed studies of decision-making to clarify experimental-operational differences between externally guided and internally guided decision-making. Second, using multi-level kernel density analysis, a whole-brain-based quantitative meta-analysis of neuroimaging studies was performed. Our meta-analysis revealed that the neural network used predominantly for internally guided decision-making differs from that for externally guided decision-making under uncertainty. This result suggests that studying only externally guided decision-making under uncertainty is insufficient to account for decision-making processes in the brain. Finally, based on the review and results of the meta-analysis, we discuss the differences and relations between decision-making of these two types in terms of their operational, neuronal, and theoretical characteristics.


ADAMTS-13 in the Diagnosis and Management of Thrombotic Microangiopathies.
Thrombotic microangiopathies (TMAs) comprise a group of distinct disorders characterized by microangiopathic hemolytic anemia, thrombocytopenia, and microvascular thrombosis. For many years distinction between these TMAs, especially between thrombotic thrombocytopenic purpura (TTP) and hemolytic uremic syndrome (HUS), remained purely clinical and hard to make. Recent discoveries shed light on different pathogenesis of TTP and HUS. Ultra-large von Willebrand factor (UL-VWF) platelet thrombi, resulting from the deficiency of cleavage protease which is now known as ADAMTS-13 (a disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13), were found to cause TTP pathology, while Shiga toxins or abnormalities in regulation of the complement system cause microangiopathy and thrombosis in HUS. TMAs may appear in various conditions such as pregnancy, inflammation, malignancy, or exposure to drugs. These conditions might cause acquired TTP, HUS, or other TMAs, or might be a trigger in individuals with genetic predisposition to ADAMTS-13 or complement factor H deficiency. Differentiation between these TMAs is highly important for urgent initiation of appropriate therapy. Measurement of ADAMTS-13 activity and anti-ADAMTS-13 antibody levels may advance this differentiation resulting in accurate diagnosis. Additionally, assessment of ADAMTS-13 levels can be a tool for monitoring treatment efficacy and relapse risk, allowing consideration of therapy addition or change. In the past few years, great improvements in ADAMTS-13 assays have been made, and tests with increased sensitivity, specificity, reproducibility, and shorter turnaround time are now available. These new assays enable ADAMTS-13 measurement in routine clinical diagnostic laboratories, which may ultimately result in improvement of TMA management.


Exploring Task-Specific Independent Standing in 3- to 5-Month-Old Infants.
Perspectives on developmental milestones suggest that an infant's ability to stand without support occurs at the age of 9-16 months. The two exploratory tasks were part of a baby swimming routine, conducted over a period of 12 weeks (24 sessions), and the aim was to examine whether young infants (mean age 97 days) improved their performance in standing as measured by prolonged time-to-stand. The data suggest that 3- to 5-month-old infants are capable of demonstrating signs of motor learning in task-specific standing. The results appear remarkable when compared to the expected age required for other forms of independent standing. The developmental process of independent standing is discussed in relation to the complex interaction between genetic and environmental factors.


Psychological treatments for eating disorders.
This review summarizes recent evidence on psychological treatments for eating disorders. Eating disorders are serious psychiatric conditions requiring evidence-based intervention. Treatments have been evaluated within each eating disorder diagnosis and across diagnoses. For adults with anorexia nervosa, no one specialist treatment has been shown to be superior. Cognitive behavioral therapy and interpersonal psychotherapy remain the most established treatments for bulimia nervosa and binge eating disorder, with stepped-care approaches showing promise and new behavioral treatments under study. Enhanced cognitive behavioral therapy has improved symptoms in adults and youth. Maudsley family-based therapy is the most established treatment for youth with anorexia nervosa and may be efficacious for youth with bulimia nervosa. Interpersonal psychotherapy for the prevention of excess weight gain may be efficacious for reducing loss of control eating and weight gain in overweight youth. Significant advances in treatments have been made, including evaluation of long-term outcomes, novel approaches, and tailored extension for specific patient profiles. However, widespread access to effective eating disorder treatments remains limited. Increasing the potency and expanding the implementation of psychological treatments beyond research settings into clinical practice has strong potential to increase access to care, thereby reducing the burden of eating disorders.


Thrombotic thrombocytopenic purpura.
Thrombotic thrombocytopenic purpura is an uncommon disorder, but it continues to be of considerable interest. The disease mechanisms are unclear and the aetiology is unknown. Perhaps most enigmatic of all, the mode of action of plasma therapy, which successfully induces remission in about two-thirds of cases, is wholly inexplicable. There are currently several areas of debate on the subject of thrombotic thrombocytopenic purpura. This paper addresses these points of contention: the definition of the disease, its distinction from haemolytic uraemic syndrome, the nature of the platelet aggregating factors in the plasma of patients with acute disease, the importance of the abnormalities of von Willebrand's factor observed in the acute and quiescent phases of the disease, the nature of the factor in normal plasma that induces remission, and the possible causes of the observed superiority of plasma exchange combined with plasma infusion, over plasma infusion alone.


Sustained activation of the mitogen-activated protein (MAP) kinase cascade may be required for differentiation of PC12 cells. Comparison of the effects of nerve growth factor and epidermal growth factor.
Stimulation of PC12 cells with nerve growth factor (NGF) increased mitogen-activated protein kinase kinase (MAPKK) activity > 20-fold after 5 min to a level that was largely sustained for at least 90 min. MAPKK activity was stimulated to a similar level by epidermal growth factor (EGF), but peaked at 2 min, declining thereafter and returning to basal levels after 60-90 min. Activation of MAPKK by either growth factor occurred prior to the activation of MAP kinase, consistent with MAPKK being the physiological activator of MAP kinase. The results demonstrate that the transient activation of MAPKK by EGF and its sustained activation by NGF underlies the transient and sustained activation of MAP kinase induced by EGF and NGF respectively. NGF or EGF induced the same two forms of MAPKK that were resolved on a Mono Q column. The Peak-1 MAPKK was activated initially and partially converted into the more acidic peak-2 MAPKK after prolonged growth-factor stimulation. The Peak-2 MAPKK was 20-fold more sensitive to inactivation by the catalytic subunit of protein phosphatase 2A. Stimulation with NGF caused a striking translocation of MAP kinase from the cytosol to the nucleus after 30 min, but not nuclear translocation of MAP kinase occurred after stimulation with EGF. The results suggest that sustained activation of the MAP kinase cascade may be required for MAP kinase to enter the nucleus, where it may initiate the gene transcription events required for neuronal differentiation of PC12 cells.


Serum estrogen and tumor-positive estrogen receptor-alpha are strong prognostic classifiers of non-small-cell lung cancer survival in both men and women.
The role of tumor estrogen receptors (ERs) and serum estrogen in lung cancer is inconclusive. We investigated the hypothesis that ERs and functional single-nucleotide polymorphisms in the estrogen biosynthesis pathway are associated with poorer lung cancer survival. Lung cancer patients (n = 305) from a National Cancer Institute-Maryland (NCI-MD) case-case cohort in the Baltimore metropolitan area were used as a test cohort. To validate, 227 cases from the NCI-MD case-control cohort and 293 cases from a Norwegian lung cancer cohort were studied. Information on demographics, tobacco and reproductive histories was collected in an interviewer-administered questionnaire. Serum estrogen, progesterone, tumor messenger RNA expression of hormone receptors and germ line DNA polymorphisms were analyzed for associations with lung cancer survival. Patients in the highest tertile of serum estrogen had worse survival in all three cohorts (P combined < 0.001). Furthermore, the variant allele of estrogen receptor alpha (ER-α) polymorphism (rs2228480) was significantly associated with increased tumor ER-α levels and worse survival in all three cohorts [hazard ratio (HR) = 2.59, 95% confidence interval (CI): 1.20- 4.01; HR = 1.76, 95% CI: 1.08-2.87 and HR = 2.85, 95% CI: 1.31-4.36). Other polymorphisms associated with lower serum estrogen correlated with improved survival. Results were independent of gender and hormone replacement therapy. We report a significant association of increased serum estrogen with poorer survival among lung cancer male and female patients. Understanding the genetic control of estrogen biosynthesis and response in lung cancer could lead to improved prognosis and therapy.


Phase I Trial of the Human Double Minute 2 Inhibitor MK-8242 in Patients With Advanced Solid Tumors.
Purpose To evaluate MK-8242 in patients with wild-type TP53 advanced solid tumors. Patients and Methods MK-8242 was administered orally twice a day on days 1 to 7 in 21-day cycles. The recommended phase II dose (RP2D) was determined on the basis of safety, tolerability, pharmacokinetics (PK), and by mRNA expression of the p53 target gene pleckstrin homology-like domain, family A, member 3 ( PHLDA3). Other objectives were to characterize the PK/pharmacodynamic (PD) relationship, correlate biomarkers with response, and assess tumor response. Results Forty-seven patients received MK-8242 across eight doses that ranged from 60 to 500 mg. Initially, six patients developed dose-limiting toxicities (DLTs): grade (G) 2 nausea at 120 mg; G3 fatigue at 250 mg; G2 nausea and G4 thrombocytopenia at 350 mg; and G3 vomiting and G3 diarrhea at 500 mg. DLT criteria were revised to permit management of GI toxicities. Dosing was resumed at 400 mg, and four additional DLTs were observed: G4 neutropenia and G4 thrombocytopenia at 400 mg and G4 thrombocytopenia (two patients) at 500 mg. Other drug-related G3 and G4 events included anemia, leukopenia, pancytopenia, nausea, hyperbilirubinemia, hypophosphatemia, and anorexia. On the basis of safety, tolerability, PK, and PD, the RP2D was established at 400 mg (15 evaluable patients experienced two DLTs). PK for 400 mg (day 7) showed Cmax 3.07 μM, Tmax 3.0 hours, t1/2 (half-life) 6.6 hours, CL/F (apparent clearance) 28.9 L/h, and Vd/F (apparent volume) 274 L. Blood PHLDA3 mRNA expression correlated with drug exposure ( R2 = 0.68; P < .001). In 41 patients with postbaseline scans, three patients with liposarcoma achieved a partial response (at 250, 400, and 500 mg), 31 showed stable disease, and eight had progressive disease. In total, 27 patients with liposarcoma had a median progression-free survival of 237 days. Conclusion At the RP2D of 400 mg twice a day, MK-8242 activated the p53 pathway with an acceptable safety and tolerability profile. The observed clinical activity (partial response and prolonged progression-free survival) provides an impetus for further study of HDM2 inhibitors in liposarcoma.


Amount of smoking independently predicts carotid artery atherosclerosis severity.
Cigarette smoking is correlated with extracranial carotid artery plaque thickness. Our aim in the present study was to determine whether the level of prior cigarette use is a significant predictor of carotid artery plaque thickness when age, history of hypertension, and history of diabetes are controlled. We studied a continuous sample of 790 patients with a history of smoking referred for diagnostic ultrasound imaging of the carotid arteries. Subjects (mean age 61 years) had an average of 51 pack-years of cigarette use. History of hypertension was present in 44% and history of diabetes in 18%. Right and left maximum carotid artery plaque thicknesses were averaged for each patient; the average of this value for all 790 subjects was 1.9 mm. In bivariate analysis, age (p less than 0.0001), pack-years (p less than 0.0001), history of hypertension (p = 0.0003), and history of diabetes (p = 0.037) were each positively associated with carotid artery plaque thickness. In multiple regression analysis, age (p less than 0.0001), pack-years (p = 0.0005), and history of hypertension (p = 0.0044) were statistically significant independent predictors of carotid artery plaque thickness, but history of diabetes (p = 0.2451) was not. In smokers, the level of cigarette use is associated with an acceleration of carotid artery atherosclerosis development. This effect is independent of age, hypertension, and diabetes. These results support smoking abatement as an important element to stroke prevention in clinical practice.


'Candidatus Phytoplasma brasiliense', a new phytoplasma taxon associated with hibiscus witches' broom disease.
Hibiscus rosa-sinensis is a valuable ornamental species widely planted in Brazil. Many plants are affected by witches' broom disease, which is characterized by excessive axillary branching, abnormally small leaves, and deformed flowers, symptoms that are characteristic of diseases attributed to phytoplasmas. A phytoplasma was detected in diseased Hibiscus by amplification of rRNA operon sequences by PCRs, and was characterized by RFLP and nucleotide sequence analyses of 16S rDNA. The collective RFLP patterns of amplified 16S rDNA differed from the patterns described previously for other phytoplasmas. On the basis of the RFLP patterns, the hibiscus witches' broom phytoplasma was classified in a new 16S rRNA RFLP group, designated group 16SrXV. A phylogenetic analysis of 16S rDNA sequences from this and other phytoplasmas identified the hibiscus witches' broom phytoplasma as a member of a distinct subclade (designated subclade xiv) of the class Mollicutes. A phylogenetic tree constructed on the basis of 16S rRNA gene sequences was consistent with the hypothesis that there was divergent evolution of hibiscus witches' broom phytoplasma and its closest relatives (members of 16S rRNA RFLP group 16SrII) from a common ancestor. On the basis of unique properties of the DNA from hibiscus witches' broom phytoplasma, it is proposed that it represents a new taxon, namely 'Candidatus Phytoplasma brasiliense'.


Human Immunodeficiency Virus Infection Impairs Th1 and Th17 Mycobacterium tuberculosis-Specific T-Cell Responses.
Human immunodeficiency virus (HIV)-infected individuals have a higher risk of developing active tuberculosis (TB) than HIV-uninfected individuals, but the mechanisms underpinning this are unclear. We hypothesized that depletion of specific components of Mycobacterium tuberculosis (Mtb)-specific CD4+ and CD8+ T-cell responses contributed to this increased risk. Mtb-specific T-cell responses in 147 HIV-infected and 44 HIV-uninfected control subjects in a TB-endemic setting in Bloemfontein, South Africa, were evaluated. Using a whole-blood flow cytometry assay, we measured expression of interferon gamma, tumor necrosis factor alpha, interleukin 2, and interleukin 17 in CD4+ and CD8+ T cells in response to Mtb antigens (PPD, ESAT-6/CFP-10 [EC], and DosR regulon-encoded α-crystallin [Rv2031c]). Fewer HIV-infected individuals had detectable CD4+ and CD8+ T-cell responses to PPD and Rv2031c than HIV-uninfected subjects. Mtb-specific T cells showed distinct patterns of cytokine expression comprising both Th1 (CD4 and CD8) and Th17 (CD4) cytokines, the latter at highest frequency for Rv2031c. Th17 antigen-specific responses to all antigens tested were specifically impaired in HIV-infected individuals. HIV-associated impairment of CD4+ and CD8+Mtb-specific T-cell responses is antigen specific, particularly impacting responses to PPD and Rv2031c. Preferential depletion of Th17 cytokine-expressing CD4+ T cells suggests this T-cell subset may be key to TB susceptibility in HIV-infected individuals.


Using High-Content Screening to Generate Single-Cell Gene-Corrected Patient-Derived iPS Clones Reveals Excess Alpha-Synuclein with Familial Parkinson's Disease Point Mutation A30P.
The generation of isogenic induced pluripotent stem cell (iPSC) lines using CRISPR-Cas9 technology is a technically challenging, time-consuming process with variable efficiency. Here we use fluorescence-activated cell sorting (FACS) to sort biallelic CRISPR-Cas9 edited single-cell iPSC clones into high-throughput 96-well microtiter plates. We used high-content screening (HCS) technology and generated an in-house developed algorithm to select the correctly edited isogenic clones for continued expansion and validation. In our model we have gene-corrected the iPSCs of a Parkinson's disease (PD) patient carrying the autosomal dominantly inherited heterozygous c.88G>C mutation in the SNCA gene, which leads to the pathogenic p.A30P form of the alpha-synuclein protein. Undertaking a PCR restriction-digest mediated clonal selection strategy prior to sequencing, we were able to post-sort validate each isogenic clone using a quadruple screening strategy prior to generating footprint-free isogenic iPSC lines, retaining a normal molecular karyotype, pluripotency and three germ-layer differentiation potential. Directed differentiation into midbrain dopaminergic neurons revealed that SNCA expression is reduced in the gene-corrected clones, which was validated by a reduction at the alpha-synuclein protein level. The generation of single-cell isogenic clones facilitates new insights in the role of alpha-synuclein in PD and furthermore is applicable across patient-derived disease models.


Effects of Prosthetic Material and Framework Design on Stress Distribution in Dental Implants and Peripheral Bone: A Three-Dimensional Finite Element Analysis.
BACKGROUND The purpose of this study was to evaluate the effects of prosthetic material and framework design on the stress within dental implants and peripheral bone using finite element analysis (FEA). MATERIAL AND METHODS A mandibular implant-supported fixed dental prosthesis with different prosthetic materials [cobalt-chromium-supported ceramic (C), zirconia-supported ceramic (Z), and zirconia-reinforced polymethyl methacrylate (ZRPMMA)-supported resin (ZP)] and different connector widths (2, 3, and 4 mm) within the framework were used to evaluate stress via FEA under oblique loading conditions. Maximum principal (smax), minimum principal (smin), and von Mises (svM) stress values were obtained. RESULTS Minimum stress values were observed in the model with a 2-mm connector width for C and ZP. The models with 3-mm and 4-mm connector widths showed higher stress values than the model with a 2-mm connector width for C (48-50%) and ZP (50-52%). Similar stress values were observed in the 3- and 4-mm models. There was no significant difference in the amount of stress with Z regardless of connector width. The Z and ZP models showed similar stress values in the 3- and 4-mm models and higher stress values than in the C model. Z, ZP, and C showed the highest stress values for the model with a 2-mm connector width. CONCLUSIONS Changes in the material and width of connectors may influence stress on cortical bone, cancellous bone, and implants. C was associated with the lowest stress values. Higher maximum and minimum principal stress values were seen in cortical bone compared to cancellous bone.


The Celera Discovery System.
The Celera Discovery System (CDS) is a web-accessible research workbench for mining genomic and related biological information. Users have access to the human and mouse genome sequences with annotation presented in summary form in BioMolecule Reports for genes, transcripts and proteins. Over 40 additional databases are available, including sequence, mapping, mutation, genetic variation, mRNA expression, protein structure, motif and classification data. Data are accessible by browsing reports, through a variety of interactive graphical viewers, and by advanced query capability provided by the LION SRS search engine. A growing number of sequence analysis tools are available, including sequence similarity, pattern searching, multiple sequence alignment and Hidden Markov Model search. A user workspace keeps track of queries and analyses. CDS is widely used by the academic research community and requires a subscription for access. The system and academic pricing information are available at http://cds.celera.com.


Comparing Sensory Information Processing and Alexithymia between People with Substance Dependency and Normal.
Sensory information processing and alexithymia are two important factors in determining behavioral reactions. Some studies explain the effect of the sensitivity of sensory processing and alexithymia in the tendency to substance abuse. Giving that, the aim of the current study was to compare the styles of sensory information processing and alexithymia between substance-dependent people and normal ones. The research method was cross-sectional and the statistical population of the current study comprised of all substance-dependent men who are present in substance quitting camps of Masal, Iran, in October 2013 (n = 78). 36 persons were selected randomly by simple randomly sampling method from this population as the study group, and 36 persons were also selected among the normal population in the same way as the comparison group. Both groups was evaluated by using Toronto alexithymia scale (TAS) and adult sensory profile, and the multivariate analysis of variance (MANOVA) test was applied to analyze data. The results showed that there are significance differences between two groups in low registration (P < 0.020, F = 5.66), sensation seeking (P < 0.050, F = 1.92), and sensory avoidance (P < 0.008, F = 7.52) as a components of sensory processing and difficulty in describing emotions (P < 0.001, F = 15.01) and difficulty in identifying emotions (P < 0.002, F = 10.54) as a components of alexithymia. However, no significant difference were found between two groups in components of sensory sensitivity (P < 0.170, F = 1.92) and external oriented thinking style (P < 0.060, F = 3.60). These results showed that substance-dependent people process sensory information in a different way than normal people and show more alexithymia features than them.


Giving Voice to Values: an undergraduate nursing curriculum project.
Among the competency standards stipulated by the Australian Nursing and Midwifery Council for graduating students are competencies in moral and ethical decision making and ethics education within professions such as nursing has traditionally focussed on these competencies, on raising ethical awareness and developing skills of analysis and reasoning. However, ethics education in tertiary settings places less emphasis on developing students' capacities to act on their values. This paper explains and explores the adoption of Dr. Mary Gentile's curriculum (the Giving Voice to Values curriculum).which specifically focuses on developing students' capacities to act on their values. The curriculum (Gentile, 2010) assists students and professionals to explore, script and rehearse responses which build upon their capacity to respond in accordance with their own values in complex workplace settings in which they face conflicts of value and belief. The paper firstly examines the theoretical underpinnings of the Giving Voice to Values (GVV) curriculum. It then presents the integration and evaluation phase of a Project inspired by the GVV methodology, using a case study approach within two areas of an undergraduate nursing curriculum. As a pilot project, this initiative has provided signposts to further curriculum development and to research pathways within the UNDA School of Nursing, by highlighting students' uncertainties regarding their own professional values, and their intense struggles to voice their values within health care contexts.


Remote optogenetic activation and sensitization of pain pathways in freely moving mice.
We report a novel model in which remote activation of peripheral nociceptive pathways in transgenic mice is achieved optogenetically, without any external noxious stimulus or injury. Taking advantage of a binary genetic approach, we selectively targeted Nav1.8(+) sensory neurons for conditional expression of channelrhodopsin-2 (ChR2) channels. Acute blue light illumination of the skin produced robust nocifensive behaviors, evoked by the remote stimulation of both peptidergic and nonpeptidergic nociceptive fibers as indicated by c-Fos labeling in laminae I and II of the dorsal horn of the spinal cord. A non-nociceptive component also contributes to the observed behaviors, as shown by c-Fos expression in lamina III of the dorsal horn and the expression of ChR2-EYFP in a subpopulation of large-diameter Nav1.8(+) dorsal root ganglion neurons. Selective activation of Nav1.8(+) afferents in vivo induced central sensitization and conditioned place aversion, thus providing a novel paradigm to investigate plasticity in the pain circuitry. Long-term potentiation was similarly evoked by light activation of the same afferents in isolated spinal cord preparations. These findings demonstrate, for the first time, the optical control of nociception and central sensitization in behaving mammals and enables selective activation of the same class of afferents in both in vivo and ex vivo preparations. Our results provide a proof-of-concept demonstration that optical dissection of the contribution of specific classes of afferents to central sensitization is possible. The high spatiotemporal precision offered by this non-invasive model will facilitate drug development and target validation for pain therapeutics.


Optical resonances of bowtie slot antennas and their geometry and material dependence.
In order to provide a guide for the design and optimization of bowtie slot antennas in the visible and near infrared spectral regime, their optical properties have been investigated with emphasis on geometry and materials. Although primarily theoretical, experimental investigations for reduced thickness cases are also included. As characterized by their field patterns, two types of resonances are discussed: plasmonic and Fabry-Pérot-like resonances. These resonance types show a linear dependence on aperture perimeter and film thickness, respectively, while showing a complementary behavior with near independence of the other respective parameter. Metal properties, as in the Drude model, are also considered. Various metals with respectively different skin depths are studied, showing a nearly linear dependence of the resonance wavelength on skin depth.


Sustainable development of water services industry.
The world population is expected to increase up to 8 billion by 2015. Most of the cities in the world are scattered around the fresh water resources. These cities and villages have their own interests and they constitute a power order. Therefore, we have been trying to harmonize these cities and villages in the usage of fresh water resources and the discharging of wastewater as well, by using an engineering means of water treatment. However, fresh water resources, which are essential to our lives, have a constant amount of circulation with a period of one week to ten days. The science and technology of the water environment should recognize the limit of fresh water resources for the sustainable development of society, because the water service industries are the essential infrastructure of the community. In order to implement an appropriate risk management it is necessary to identify the goal of environmental management considering the social, natural and economical conditions. The energy-dependent technologies should be reconsidered to consume less energy and undertake more resource conservation engineering. Water science and technology cannot supply all the answers, therefore more comprehensive water management systems should be developed by other sectors, such as the food production industry.


The excreted microbiota of bats: evidence of niche specialisation based on multiple body habitats.
Animal-associated microbiotas form complex communities, which play crucial functions for their host, including susceptibility to infections. Despite increasing attention to bats as reservoirs of zoonotic pathogens, their microbiota is poorly documented, especially for samples potentially implicated in pathogen transmission such as urine and saliva. Here, using low-biomass individual samples, we examined the composition and structure of bacterial communities excreted by insectivorous bats, focusing on three body habitats (saliva, urine and faeces). We show that niche specialisation occurs as bacterial community composition was distinct across body habitats with the majority of phylotypes being body habitat specific. Our results suggest that urine harbours more diverse bacterial communities than saliva and faeces and reveal potentially zoonotic bacteria such as Leptospira, Rickettsia, Bartonella and Coxiella in all body habitats. Our study emphasised that, in addition to the traditional use of gut-associated samples such as faeces, both urine and saliva are also of interest because of their diverse microbiota and the potential transmission of pathogenic bacteria. Our results represent a critical baseline for future studies investigating the interactions between microbiota and infection dynamics in bats.


Improved RNA modification mapping of cellular non-coding RNAs using C- and U-specific RNases.
Locating ribonucleoside modifications within an RNA sequence requires digestion of the RNA into oligoribonucleotides of amenable size for subsequent analysis by LC-MS (liquid chromatography-mass spectrometry). This approach, widely referred to as RNA modification mapping, is facilitated through ribonucleases (RNases) such as T1 (guanosine-specific), U2 (purine-selective) and A (pyrimidine-specific) among others. Sequence coverage by these enzymes depends on positioning of the recognized nucleobase (such as guanine or purine or pyrimidine) in the sequence and its ribonucleotide composition. Using E. coli transfer RNA (tRNA) and ribosomal RNA (rRNA) as model samples, we demonstrate the ability of complementary nucleobase-specific ribonucleases cusativin (C-specific) and MC1 (U-specific) to generate digestion products that facilitate confident mapping of modifications in regions such as G-rich and pyrimidine-rich segments of RNA, and to distinguish C to U sequence differences. These enzymes also increase the number of oligonucleotide digestion products that are unique to a specific RNA sequence. Further, with these additional RNases, multiple modifications can be localized with high confidence in a single set of experiments with minimal dependence on the individual tRNA abundance in a mixture. The sequence overlaps observed with these complementary digestion products and that of RNase T1 improved sequence coverage to 75% or above. A similar level of sequence coverage was also observed for the 2904 nt long 23S rRNA indicating their utility has no dependence on RNA size. Wide-scale adoption of these additional modification mapping tools could help expedite the characterization of modified RNA sequences to understand their structural and functional role in various living systems.


Immediate Reconstruction of Amputated Ear With Fascia Flap.
Various surgical methods have been used for reconstruction of an amputated ear, including microsurgical replantation and delayed reconstruction using synthetic material or autologous rib cartilage. The authors share our experience of immediate reconstruction using amputated cartilage and discuss the advantages compared to other techniques of reconstruction. The authors retrospectively reviewed the medical records of 3 patients who underwent immediate reconstruction of amputated ear by a single operator. In the cases, the amputees were washed; the skin and cartilage were separated. The ear cartilage was fixed to its original position with non-absorbable suture. Regional fascia flap was elevated and covered the ear cartilage. The amputee skin was redraped to cover the fascia flap. Several months after the operation, photographic assessment was done. All 3 cases showed well-defined convolution, tolerable skin color, and patient satisfaction without any major complications. A patient showed mild temporal incision site alopecia. The above immediate reconstruction method can achieve reliable and favorable result with minimal complications.


Copayment reductions generate greater medication adherence in targeted patients.
A large value-based insurance design program offered by Blue Cross Blue Shield of North Carolina eliminated generic medication copayments and reduced copayments for brand-name medications. Our study showed that the program improved adherence to medications for diabetes, hypertension, hyperlipidemia, and congestive heart failure. We found that adherence improved for enrollees, ranging from a gain of 3.8 percentage points for patients with diabetes to 1.5 percentage points for those taking calcium-channel blockers, when compared to others whose employers did not offer a similar program. An examination of longer-term adherence and trends in health care spending is still needed to provide a compelling evidence base for value-based insurance design.


Therapy of rheumatoid arthritis: new developments and trends.
The medical therapy of rheumatoid arthritis (RA) has been influenced strongly during the past decade by the recognition that many patients develop joint damage within the first year of disease. This observation has motivated rheumatologists to initiate disease-modifying antirheumatic drugs (DMARDs) early in the disease course. This trend has been matched by the increased use of combination DMARD therapy, with the aim to maximize control of the signs and symptoms of RA and limit the development of joint damage. The results from controlled clinical trials generally have supported the concept that early, aggressive treatment with DMARDs is superior to less intensive strategies. In addition, certain DMARD combinations are more effective than the individual components of these regimens, but the evidence is strong for only a few DMARD combinations such as methotrexate and cyclosporine A. Three new drugs have been recently approved for the treatment of RA. Celecoxib, a selective cyclooxygenase-2 inhibitor, has similar clinical efficacy as conventional nonsteroidal anti-inflammatory drugs, and in short-term studies causes no more gastric and duodenal ulcers and erosions than patients treated with placebo. Treatment with leflunomide, an inhibitor of pyrimidine synthesis, has been shown in controlled clinical trials to produce significant clinical improvement in 50% to 60% of patients with RA and delay radiologic progression of disease. The era of biologic therapy has dawned with the apparent success of tumor necrosis factor (TNF)-alpha blockade using etanercept, a recombinant TNF receptor:Fc fusion protein, and infliximab, a chimeric anti-TNF monoclonal antibody. These new agents expand our treatment options in RA and should lead to innovative and more effective treatment approaches.


Patients use fewer analgesics following supervised exercise therapy and patient education: an observational study of 16 499 patients with knee or hip osteoarthritis.
To investigate changes in analgesic use before and after supervised exercise therapy and patient education in patients with knee or hip osteoarthritis (OA). We recruited 16 499 of 25 933 eligible patients (64%; mean age 64.9; SD 9.6; 73% women) from the Good Life with osteoArthritis in Denmark (GLA:D) registry. Change in proportions of analgesic users (categorised according to analgesic risk profile; opioids > non-steroidal anti-inflammatory drugs > paracetamol) was assessed from before to after an 8-week supervised exercise therapy and patient education programme targeting knee or hip OA pain and functional limitations. Patients reported 13.2 mm (95% CI 12.8 to 13.6) less pain (visual analogue scale 0-100 mm) at follow-up compared with baseline. The proportion of analgesic users reduced from 62.2% (95% CI 61.5 to 63.0) at baseline to 44.1% (95% CI 43.3 to 44.9) at follow-up (absolute change: 18.1% (95% CI 17.3 to 19.0)). Among patients using analgesics at baseline, 52% changed to a lower risk analgesic or discontinued analgesic use. The proportion of opioid users after the exercise therapy was 2.5% (95% CI 2.1 to 2.9) lower than baseline; this represents a relative reduction of 36%. Among patients with knee or hip OA using analgesics, more than half either discontinued analgesic use or shifted to lower risk analgesics following an 8-week structured exercise therapy and patient education programme (GLA:D). These data encourage randomised controlled trial evaluation of whether supervised exercise therapy, combined with patient education, can reduce analgesic use, including opioids, among patients with knee and hip OA pain.


Adapting a weight management tool for Latina women: a usability study of the Veteran Health Administration's MOVE!23 tool.
Obesity disproportionately affects Latina women, but few targeted, technology-assisted interventions that incorporate tailored health information exist for this population. The Veterans Health Administration (VHA) uses an online weight management tool (MOVE!23) which is publicly available, but was not designed for use in non-VHA populations. We conducted a qualitative study to determine how interactions between the tool and other contextual elements impacted task performance when the target Latina users interacted with MOVE!23. We sought to identify and classify specific facilitators and barriers that might inform design changes to the tool and its context of use, and in turn promote usability. Six English-speaking, adult Latinas were recruited from an inner city primary care clinic and a nursing program at a local university in the United States to engage in a "Think-Aloud" protocol while using MOVE!23. Sessions were recorded, transcribed, and coded to identify interactions between four factors that contribute to usability (Tool, Task, User, Context). Five themes influencing usability were identified: Technical Ability and Technology Preferences; Language Confusion and Ambiguity; Supportive Tool Design and Facilitator Guidance; Relevant Examples; and Personal Experience. Features of the tool, task, and other contextual factors failed to fully support participants at times, impeding task completion. Participants interacted with the tool more readily when its language was familiar and content was personally relevant. When faced with ambiguity and uncertainty, they relied on the tool's visual cues and examples, actively sought relevant personal experiences, and/or requested facilitator support. The ability of our participants to successfully use the tool was influenced by the interaction of individual characteristics with those of the tool and other contextual factors. We identified both tool-specific and context-related changes that could overcome barriers to the use of MOVE!23 among Latinas. Several general considerations for the design of eHealth tools are noted.


Characterization of secretory protein translocation: ribosome-membrane interaction in endoplasmic reticulum.
Secretory proteins are synthesized on ribosomes bound to the membrane of the endoplasmic reticulum (ER). After the selection of polysomes synthesizing secretory proteins and their direction to the membrane of the ER via signal recognition particle (SRP) and docking protein respectively, the polysomes become bound to the ER membrane via an unknown, protein-mediated mechanism. To identify proteins involved in protein translocation, beyond the (SRP-docking protein-mediated) recognition step, controlled proteolysis was used to functionally inactivate rough microsomes that had previously been depleted of docking protein. As the membranes were treated with increasing levels of protease, they lost their ability to be functionally reconstituted with the active cytoplasmic fragment of docking protein (DPf). This functional inactivation did not correlate with a loss of either signal peptidase activity, nor with the ability of the DPf to reassociate with the membrane. It did correlate, however, with a loss of the ability of the microsomes to bind ribosomes. Ribophorins are putative ribosome-binding proteins. Immunoblots developed with monoclonal antibodies against canine ribophorins I and II demonstrated that no correlation exists between the protease-induced inability to bind ribosomes and the integrity of the ribophorins. Ribophorin I was 85% resistant and ribophorin II 100% resistant to the levels of protease needed to totally eliminate ribosome binding. Moreover, no direct association was found between ribophorins and ribosomes; upon detergent solubilization at low salt concentrations, ribophorins could be sedimented in the presence or absence of ribosomes. Finally, the alkylating agent N-ethylmaleimide was shown to be capable of inhibiting translocation (beyond the SRP-docking protein-mediated recognition step), but had no affect on the ability of ribosomes to bind to ER membranes. We conclude that potentially two additional proteinaceous components, as yet unidentified, are involved in protein translocation. One is protease sensitive and possibly involved in ribosome binding, the other is N-ethylmaleimide sensitive and of unknown function.


Intrapartum temperature elevation, epidural use, and adverse outcome in term infants.
To examine the association of intrapartum temperature elevation with adverse neonatal outcome among low-risk women receiving epidural analgesia and evaluate the association of epidural with adverse neonatal outcome without temperature elevation. We studied all low-risk nulliparous women with singleton pregnancies ≥37 weeks delivering at our hospital during 2000, excluding pregnancies where infants had documented sepsis, meningitis, or a major congenital anomaly. Neonatal outcomes were compared between women receiving (n = 1538) and not receiving epidural analgesia (n = 363) in the absence of intrapartum temperature elevation (≤99.5°F) and according to the level of intrapartum temperature elevation within the group receiving epidural (n = 2784). Logistic regression was used to evaluate neonatal outcome while controlling for confounders. Maternal temperature >100.4°F developed during labor in 19.2% (535/2784) of women receiving epidural compared with 2.4% (10/425) not receiving epidural. In the absence of intrapartum temperature elevation (≤99.5°F), no significant differences were observed in adverse neonatal outcomes between women receiving and not receiving epidural. Among women receiving epidural, a significant linear trend was observed between maximum maternal temperature and all neonatal outcomes examined including hypotonia, assisted ventilation, 1- and 5-min Apgar scores <7, and early-onset seizures. In regression analyses, infants born to women with fever >101°F had a two- to sixfold increased risk of all adverse outcomes examined. The proportion of infants experiencing adverse outcomes increased with the degree of epidural-related maternal temperature elevation. Epidural use without temperature elevation was not associated with any of the adverse outcomes we studied.


Separation, Aspiration, and Fat Equalization: SAFE Liposuction Concepts for Comprehensive Body Contouring.
Separation, aspiration, and fatty equilibration (SAFE) liposuction uses a process approach to body contouring and minimizes injury to surrounding structures. The multistep process allows for (1) fat separation, (2) lipoaspiration, and (3) fat equalization. The purpose of this study was to review both outcomes and complications of primary SAFE liposuction. Retrospective chart review was completed of patients undergoing SAFE liposuction from January of 2006 to January of 2011. Patient selection was limited to those undergoing liposuction alone with no adjuvant excisional procedures. Data were collected regarding demographics, body mass index, operative details, and outcomes. Seven hundred thirty-four patients were identified as having undergone SAFE liposuction. One hundred twenty-nine patients were found to have been treated with liposuction alone. Patient age ranged from 18 to 42 years and body mass index ranged from 18 to 42 kg/m (mean, 26.3 kg/m). Seven patients (5.4 percent) underwent treatment of the face and neck, six patients (4.7 percent) underwent treatment of upper extremities, 13 patients (10.1 percent) underwent treatment of the chest, 20 patients (15.5 percent) underwent treatment of lower extremities, 32 patients (24.8 percent) underwent treatment of the circumferential trunk, and 51 patients (39.5 percent) underwent treatment of circumferential trunk and additional area(s). No major complications occurred. Five of the 129 patients (3.87 percent) developed the minor complication of seroma formation. SAFE liposuction is a multistep process approach to body contouring consisting of (1) fat separation, (2) lipoaspiration, and (3) fat equalization. The results of this study show such technique to be safe and effective. Therapeutic, IV.


Pre-school screening for developmental and emotional health: Comparison with neurodevelopmental assessment.
The study aim was to compare detection of and referral for developmental and emotional problems in a school readiness screening programme (New Zealand Before School Check, B4SC) with that of a comprehensive neurodevelopmental assessment. This is a prospective cohort study of children (n = 274) born at risk of neonatal hypoglycaemia and recruited to a follow-up study of neurodevelopmental outcomes at 4.5 years (Children with Hypoglycaemia and their Later Development (CHYLD) Study). Children identified as of significant concern for developmental and emotional problems, and referrals made, were compared in the B4SC and CHYLD Study. Scores of the parent-completed Strengths and Difficulties Questionnaire used in both assessments were compared. Of the 274 children who underwent clinical neurodevelopmental assessment at a mean (standard deviation) age of 53.3 (1.8) months, 237 had the B4SC developmental and emotional health screening. Of these, 44 (19%) children met B4SC referral criteria, and 15 (6%) were referred, but only 21 (9%) children met CHYLD referral criteria, and 10 (4%) were referred. Twelve children (5%) met both the B4SC and CHYLD referral criteria, and two were referred by both. When assessed twice, 39 (17%) children changed parent-completed Strengths and Difficulties Questionnaire category. Children who did not have B4SC screening had higher mean total difficulties score (10.5 vs. 8.2, P = 0.009) and were more likely to have cognitive delay than those who were screened (19% vs. 8%, P = 0.04). More children met referral criteria for the B4SC screening programme than for a more comprehensive neurodevelopmental assessment. Children who did not have screening had a higher incidence of cognitive and behaviour problems than those who did.


In-silico model of skin penetration based on experimentally determined input parameters. Part II: mathematical modelling of in-vitro diffusion experiments. Identification of critical input parameters.
This work describes a framework for in-silico modelling of in-vitro diffusion experiments illustrated in an accompanying paper [S. Hansen, A. Henning, A. Naegel, M. Heisig, G. Wittum, D. Neumann, K.-H. Kostka, J. Zbytovska, C.M. Lehr, U.F. Schaefer, In-silico model of skin penetration based on experimentally determined input parameters. Part I: experimental determination of partition and diffusion coefficients, Eur. J. Pharm. Biopharm. 68 (2008) 352-367 [corrected] A mathematical model of drug permeation through stratum corneum (SC) and viable epidermis/dermis is presented. The underlying geometry for the SC is of brick-and-mortar character, meaning that the corneocytes are completely embedded in the lipid phase. The geometry is extended by an additional compartment for the deeper skin layers (DSL). All phases are modelled with homogeneous diffusivity. Lipid-donor and SC-DSL partition coefficients are determined experimentally, while corneocyte-lipid and DSL-lipid partition coefficients are derived consistently with the model. Together with experimentally determined apparent lipid- and DSL-diffusion coefficients, these data serve as direct input for computational modelling of drug transport through the skin. The apparent corneocyte diffusivity is estimated based on an approximation, which uses the apparent SC- and lipid-diffusion coefficients as well as corneocyte-lipid partition coefficients. The quality of the model is evaluated by a comparison of concentration-SC-depth-profiles of the experiment with those of the simulation. Good agreements are obtained, and by an analysis of the underlying model, critical parameters of the models can be identified more easily.


A prominent β-hairpin structure in the winged-helix domain of RECQ1 is required for DNA unwinding and oligomer formation.
RecQ helicases have attracted considerable interest in recent years due to their role in the suppression of genome instability and human diseases. These atypical helicases exert their function by resolving a number of highly specific DNA structures. The crystal structure of a truncated catalytic core of the human RECQ1 helicase (RECQ1(49-616)) shows a prominent β-hairpin, with an aromatic residue (Y564) at the tip, located in the C-terminal winged-helix domain. Here, we show that the β-hairpin is required for the DNA unwinding and Holliday junction (HJ) resolution activity of full-length RECQ1, confirming that it represents an important determinant for the distinct substrate specificity of the five human RecQ helicases. In addition, we found that the β-hairpin is required for dimer formation in RECQ1(49-616) and tetramer formation in full-length RECQ1. We confirmed the presence of stable RECQ1(49-616) dimers in solution and demonstrated that dimer formation favours DNA unwinding; even though RECQ1 monomers are still active. Tetramers are instead necessary for more specialized activities such as HJ resolution and strand annealing. Interestingly, two independent protein-protein contacts are required for tetramer formation, one involves the β-hairpin and the other the N-terminus of RECQ1, suggesting a non-hierarchical mechanism of tetramer assembly.


The Association Between Coffee Consumption and Local Anesthesia Failure: Social Beliefs and Scientific Evidence.
Introduction In our clinical practice, we have encountered patients who reported the failure of local anesthesia due to excessive coffee consumption and required higher-than-normal doses of local anesthesia. Therefore, our study aimed to assess the awareness and knowledge of coffee consumption, its effect on local anesthesia, and the available scientific evidence among the public, patients, and clinicians in dental practice. Material and Methods A cross-sectional survey with two sets of questionnaires was designed based on the Likert scale. A 5-point scale was used to assess agreement and frequency. Yes/no and open-ended questions were used for the assessment. Questionnaires were distributed among the clinicians, patients, and the public. Data were analyzed with descriptive linear statistics. Results Of the 430 responses provided by patients and the general public, more than 40% believed that the local anesthetic failure was caused by excessive coffee consumption. Among the 235 responses provided by the clinicians, 65% of the clinicians reported encountering patients with local anesthesia failure and believed it could be due to excessive coffee consumption. However, only 9% of the clinicians were aware of scientific evidence regarding the effect of coffee consumption on local anesthesia failure. Conclusion Surprisingly, the majority of clinicians believed that caffeine had an effect on the reduction of local anesthesia; however, only a few of them had scientific knowledge. The available scientific evidence relates that caffeine can influence cognitive performance by increasing alertness, as well as sleep deprivation causing stress and anxiety, which partially explains the local anesthetic failure among coffee consumers. Therefore, a stress reduction protocol should be a routine daily practice for a dentist to reduce the failure rate of local anesthesia.


Regioselective Synthesis of Functionalized 3- or 5-Fluoroalkyl Isoxazoles and Pyrazoles from Fluoroalkyl Ynones and Binucleophiles.
A facile synthetic route toward either 3- or 5-fluoroalkyl-substituted isoxazoles or pyrazoles containing an additional functionalization site was developed and applied on a multigram scale. The elaborated approach extends the scope of fluoroalkyl substituents for introduction into the heterocyclic moiety, and uses convenient transformations of the side chain for incorporation of fluoroalkyl-substituted azoles into the structures of biologically active molecules. The utility of the obtained building blocks for isosteric replacement of alkyl-substituted isoxazole and pyrazole was shown by the synthesis of fluorinated Isocarboxazid and Mepiprazole analogues.


Methods for exploring the semantics of the relationships between co-occurring UMLS concepts.
To characterize the relationships among UMLS concepts that co-occur as MeSH descriptors in MEDLINE citations (1990-1999). 18,485 UMLS concepts involved in 7,928,608 directed pairs of co-occurring concepts were studied. For each directed pair of concepts C1-C2: (i) the "family" of C1 was built, using the UMLS Metathesaurus, and we tested whether or not C2 belonged to C1's family; (ii) we used the semantic categorization of Metathesaurus concepts through the UMLS Semantic Network and Semantic Groups to represent the semantics of the relationships between C1 and C2. In 6.5% of the directed pairs, the co-occurring concept C2 was found within the "family" of C1. Detailed results are given. The most frequent co-occurrences involved "Chemicals and Drugs" and "Chemicals and Drugs", as well as "Disorders" and "Chemicals and Drugs". This work takes advantage of both symbolic and statistical information represented in the UMLS, and analyzes their overlap. Further research is suggested.


[Risk factors for Parkinson disease: an epidemiologic study].
The etiology of Parkinson's disease (PD) remains in a certain part unknown. Both genetic susceptibility and environmental factors are sometimes considered to be putative contributors to its origin. Recent epidemiologic studies have focused on the possible role of environmental risk factors present during adult life or aging, once pure genetic forms of PD are rare. The purpose of this study was to investigate possible environmental and familial risk factors for PD. We performed a hospital based case-control study using 88 PD patients with neurologist confirmed diagnostic, and 176 sex, age, and residence similar controls. Several possible risk factors were evaluated related to life style, past history, family history, occupational history and other exposures to potential neurotoxin agents. Statistical differences, using a 95% confidence interval, were observed in positive family history of PD (p = 0,002), occupation category (p = 0,001), rural living (p = 0,037), living/working near a industry (p = 0,017), exposure to pesticides, herbicides and in-secticides (p < 0,001), coffee consumption (p = 0,036) and tea consumption (p = 0,001). Sex and age adjusted logistic regression showed as potential risk factors, a positive family history of PD (odds ratio [OR] = 9,996; 95% confidence interval [CI] = 2,19-45,597), blue collar occupations (OR = 3,967; 95% CI = 1,670-9,426), exposure to pesticides, herbicides and insecticides (OR = 2,619 ; 95% CI = 1,170-5,862). An inverse relationship was found between tea consumption and the risk of PD (OR = 0,356; 95% CI = 0,174-0,727). The results of the study show that both familial and environmental factors may contribute to the development of PD. Like other studies suggest, PD is of unknown, but presumably multifactorial etiology.


miR-19 is a key oncogenic component of mir-17-92.
Recent studies have revealed the importance of multiple microRNAs (miRNAs) in promoting tumorigenesis, among which mir-17-92/Oncomir-1 exhibits potent oncogenic activity. Genomic amplification and elevated expression of mir-17-92 occur in several human B-cell lymphomas, and enforced mir-17-92 expression in mice cooperates with c-myc to promote the formation of B-cell lymphomas. Unlike classic protein-coding oncogenes, mir-17-92 has an unconventional gene structure, where one primary transcript yields six individual miRNAs. Here, we functionally dissected the individual components of mir-17-92 by assaying their tumorigenic potential in vivo. Using the Emu-myc model of mouse B-cell lymphoma, we identified miR-19 as the key oncogenic component of mir-17-92, both necessary and sufficient for promoting c-myc-induced lymphomagenesis by repressing apoptosis. The oncogenic activity of miR-19 is at least in part due to its repression of the tumor suppressor Pten. Consistently, miR-19 activates the Akt-mTOR (mammalian target of rapamycin) pathway, thereby functionally antagonizing Pten to promote cell survival. Our findings reveal the essential role of miR-19 in mediating the oncogenic activity of mir-17-92, and implicate the functional diversity of mir-17-92 components as the molecular basis for its pleiotropic effects during tumorigenesis.


Is transferring an educational innovation actually a process of transformation?
Recent debates question the extent to which adopting an educational innovation requires compromise between the innovation's original design and the adoption site's context. Through compromises, the innovation's fundamental principles may be transferred, transformed, or abandoned. This paper analyzes such compromises during the piloting of Team-Based Learning (TBL). We ask: When is the process of transferring an innovation actually a process of transformation? This study is an autoethnography of our research team's implementation process. Autoethnographies are personalized accounts where authors draw on their own experiences to extend understanding of a particular topic. To conduct this autoethnography, we used an in-depth, interactive interview with the piloting clinician educator. In the analysis of TBL's fundamental principles, some aspects of the principles transferred easily, while others were transformed. Analysis raised concerns that the transformations threatened the foundational principles of TBL. While an educational innovation's techniques may seem to be surface structures, they are realizations of deeper fundamental principles. The fundamental principles are themselves realizations of the innovation's foundational philosophy. When techniques and/or principles are modified to a context, it is important to analyze if the modifications continue to uphold the innovation's philosophy.


Pseudomonas spp. diversity is negatively associated with suppression of the wheat take-all pathogen.
Biodiversity and ecosystem functioning research typically shows positive diversity- productivity relationships. However, local increases in species richness can increase competition within trophic levels, reducing the efficacy of intertrophic level population control. Pseudomonas spp. are a dominant group of soil bacteria that play key roles in plant growth promotion and control of crop fungal pathogens. Here we show that Pseudomonas spp. richness is positively correlated with take-all disease in wheat and with yield losses of ~3 t/ha in the field. We modeled the interactions between Pseudomonas and the take-all pathogen in abstract experimental microcosms, and show that increased bacterial genotypic richness escalates bacterial antagonism and decreases the ability of the bacterial community to inhibit growth of the take-all pathogen. Future work is required to determine the generality of these negative biodiversity effects on different media and directly at infection zones on root surfaces. However, the increase in competition between bacteria at high genotypic richness and the potential loss of fungal biocontrol activity highlights an important mechanism to explain the negative Pseudomonas diversity-wheat yield relationship we observed in the field. Together our results suggest that the effect of biodiversity on ecosystem functioning can depend on both the function and trophic level of interest.


Dynamic dissolution testing to establish in vitro/in vivo correlations for montelukast sodium, a poorly soluble drug.
The objectives of the study was to develop a dissolution test method that can be used to predict the oral absorption of montelukast sodium, and to establish an in vitro/in vivo correlation (IVIVC) using computer simulations. Drug solubility was measured in different media. The dissolution behaviour of montelukast sodium 10 mg film coated tablets was studied using the flow-through cell dissolution method following a dynamic pH change protocol, as well as in the USP Apparatus 2. Computer simulations were performed using GastroPlus. Biorelevant dissolution media (BDM) prepared using bile salts and lecithin in buffers was used as the dissolution media, as well as the USP simulated intestinal fluid (SIF) pH 6.8 and blank FaSSIF pH 6.5. Dissolution tests in the USP Apparatus 2 were performed under a constant pH condition, while the pH range used in the flow through cells was pH 2.0 to 7.5. The in vitro data were used as input functions into GastroPlus to simulate the in vivo profiles of the drug. The solubility of montelukast sodium was low at low pH, but increased as the pH was increased. There was no significant difference in solubility in the pH range of 5.0 to 7.5 in blank buffers, but the drug solubility was higher in biorelevant media compared with the corresponding blank buffers at the same pH. Using the flow through cells, the dissolution rate was fast in simulated gastric fluid containing 0.1% SLS. The dissolution rate slowed down when the medium was changed to FaSSIF pH 6.5 and increased when the medium was changed to FaSSIF medium at pH 7.5. In the USP Apparatus 2, better dissolution was observed in FaSSIF compared with the USP buffers and blank FaSSIF with similar pH values. Dissolution was incomplete with less than 10% of the drug dissolved in the USP-SIF, and was practically non existent in blank FaSSIF pH 6.5. The in vitro results of the dynamic dissolution test were able to predict the clinical data from a bioavailability study best. Dynamic dissolution testing using the flow through cell seems to be a powerful tool to establish in vitro/in vivo correlations for poorly soluble drugs as input function into GastroPlus.