Dominant cross-reactive antibodies generated during the response to a variety of oral bacterial species detect phosphorylcholine.
The intraperitoneal immunization of Balb/c mice with subgingival plaque from advanced periodontal pockets or with certain strains of Actinobacillus actinomycetemcomitans, Fusobacterium nucleatum, Actinomyces israelii, Streptococcus mitis, or Streptococcus oralis yielded frequently indistinguishable IgM monoclonal antibodies which were reactive with antigens from a variety of oral bacteria. This study aimed to characterize the specificity of such monoclonal antibodies and the diversity of oral bacteria expressing this target antigen or epitope. Using a competitive enzyme-linked immunosorbent assay to study a variety of competitor substances for their capacity to bind to the monoclonal antibodies, we identified phosphorylcholine as the recognized epitope. The concentration of positive bacteria with extraordinarily bright cell wall fluorescence in indirect immunofluorescence assays varied between 0.1% and 15% in subgingival and from 10 to 40% in supragingival plaque samples. Labeled bacteria belonged to different morphotypes, including cocci, rods, and filaments. Of 75 species tested in vitro, 14 gram-positive and four gram-negative species were found to harbor positive strains. Haemophilus aphrophilus, Streptococcus mitis, Actinomyces georgiae, Actinomyces gerencseriae, Actinomyces israelii, and Actinomyces odontolyticus were human oral species of which all tested strains were capable of binding the cross-reactive monoclonal antibodies. In contrast, Actinomyces naeslundii was consistently negative. These data provide evidence for a much more common expression of phosphorylcholine by oral bacteria than hitherto believed but do not indicate an obvious association of phosphorylcholine expression with oral health or inflammatory periodontal diseases.


Sensorineural hearing loss in people with deletions of 18q: hearing in 18q-.
The objective of this study was to characterize hearing loss in individuals with deletions of distal chromsome18q and to identify the smallest region of overlap of their deletions, thereby identifying potential causative genes. The clinical data were collected via a retrospective case study. Molecular data were obtained via high-resolution chromosome microarray analysis. The study was conducted as a component of the ongoing research protocols at the Chromosome 18 Clinical Research Center at the University of Texas Health Science Center at San Antonio. Thirty-eight participants with a deletion of the distal portion of the long arm of chromosome 18 were recruited to this study. The participants underwent an otologic examination as well as a basic audiometry evaluation. Blood samples were obtained, and high-resolution chromosome microarray analysis was performed. Pure tone averages and speech discrimination scores were determined for each participant. The region of hemizygosity for each participant was determined to within 2 Kb each of their breakpoints. Twenty-four participants (63%) had high-frequency hearing loss, similar to the pattern seen in presbycusis. Comparison of microarray results allowed identification of eight genes, including the candidate gene for dysmyelination (MBP). Individuals with a deletion of a 2.8 Mb region of 18q23 have a high probability (83%) of high-frequency sensorineural hearing loss.


High prevalence of cyclovirus Vietnam (CyCV-VN) in plasma samples from Madagascan healthy blood donors.
Cycloviruses, small ssDNA viruses belonging to the Circoviridae family, have been suggested as possible causes of enteric, respiratory and neurological disorders in human patients. One of these species, cyclovirus-Vietnam (CyCV-VN), initially isolated from cerebrospinal fluid samples of patients with unexplained neurological disorders, has since been reported in serum samples from chronically patients infected with HBV, HCV or HIV, in Italy. On the other hand, CyCV-VN was not detected in serum samples from healthy individuals. Here, we report on a high prevalence of 43.4% (40/92) of CyCV-VN in plasma samples from asymptomatic blood donors from Madagascar. Interestingly, this virus was not detected by metagenomics and PCR in six other African countries, suggesting regional differences in CyCV-VN prevalence across Africa. Phylogenetic analysis based on the complete genomes showed that CyCV-VN sequences isolated from blood were most closely related to sequences previously reported from human stool in Madagascar. Further investigations using larger cohorts are required to determine the global epidemiology, the natural history and the pathological significance, if any, of CyCV-VN infection in humans.


Practical discourse in nursing: going beyond empiricism and historicism.
This discussion presents a critical appraisal of recent works in Advances in Nursing Science that have dealt with metatheoretical concerns. In metatheoretical analysis, such as that presented by Silva and Rothbart, nurses are beginning to focus on the traditions that have guided research. This process should be extended by reviewing historical stages of development in empiricist and postempiricist philosophy. Recent work in postempiricist philosophy is especially important since it can help nursing transcend historicist insights. Continental philosophy goes beyond historicist critique by focusing on practical discourse within a community of investigators. This is a stage of development that can be rationally decisive for the scientific development of nursing.


Gallic acid improves glucose tolerance and triglyceride concentration in diet-induced obesity mice.
Gallic acid, a phenolic phytochemical, has been shown to exert a variety of effects, including anti-oxidative, anti- carcinogenic, anti-allergic, and anti-inflammatory effects. In this study, we attempted to determine whether gallic acid affects metabolic syndrome such as obesity and diabetes. Diet-induced obesity mice were treated intraperitoneally once per day with gallic acid (10 mg/kg/day). After 2 weeks of treatment, the mice were sacrificed to collect the blood for metabolic parameter assessments, and the adipose tissues and liver to weigh and analyze. The triglyceride concentrations were significantly improved in the gallic acid group relative to those measured in the control group. And most importantly, the blood glucose concentrations in the gallic acid group were significantly improved. In the epididymal white adipose tissue of the gallic acid group, adipocyte size was reduced, PPARγ expression was induced, and the Akt signaling pathway was activated. Our results demonstrate that gallic acid improves glucose tolerance and lipid metabolism in the obesity mice, thereby showing evidence of anti-hyperglycemic activity. The findings of an upregulation of PPARγ expression and Akt activation also contribute to our current understanding of the mechanisms underlying the effects of gallic acid on glucose metabolism.


An aqueous H+ permeation pathway in the voltage-gated proton channel Hv1.
Hv1 voltage-gated proton channels mediate rapid and selective transmembrane H(+) flux and are gated by both voltage and pH gradients. Selective H(+) transfer in membrane proteins is commonly achieved by Grotthuss proton 'hopping' in chains of ionizable amino acid side chains and intraprotein water molecules. To identify whether ionizable residues are required for proton permeation in Hv1, we neutralized candidate residues and measured expressed voltage-gated H(+) currents. Unexpectedly, charge neutralization was insufficient to abrogate either the Hv1 conductance or coupling of pH gradient and voltage-dependent activation. Molecular dynamics simulations revealed water molecules in the central crevice of Hv1 model structures but not in homologous voltage-sensor domain (VSD) structures. Our results indicate that Hv1 most likely forms an internal water wire for selective proton transfer and that interactions between water molecules and S4 arginines may underlie coupling between voltage- and pH-gradient sensing.


Differences in self- and other-induced Mu suppression are correlated with empathic abilities.
Recent research suggests a role of the human mirror neuron system in empathic processing. Electroencephalographic (EEG) mu suppression in the 8-13Hz band-range has been proposed to reflect mirror-like activation of sensorimotor cortices. We therefore investigated the relationship between suppression in the 8-13Hz range and empathic abilities as measured by the Interpersonal Reactivity Index (IRI). Based on previous work showing significant differences between execution- and observation-related mu suppression and on theories of empathy pointing to the importance of discriminating self from other, we predicted that the size of this difference within individuals would correlate with participants' perspective-taking abilities. Larger differences in execution- and observation-induced mu suppression were associated with greater scores on the perspective-taking subscale of the IRI. The current data are therefore consistent with the claim that neural activity reflected in mu suppression is related to empathic abilities.


TrioGEF1 controls Rac- and Cdc42-dependent cell structures through the direct activation of rhoG.
Rho GTPases regulate the morphology of cells stimulated by extracellular ligands. Their activation is controlled by guanine exchange factors (GEF) that catalyze their binding to GTP. The multidomain Trio protein represents an emerging class of &Rgr; regulators that contain two GEF domains of distinct specificities. We report here the characterization of Rho signaling pathways activated by the N-terminal GEF domain of Trio (TrioD1). In fibroblasts, TrioD1 triggers the formation of particular cell structures, similar to those elicited by RhoG, a GTPase known to activate both Rac1 and Cdc42Hs. In addition, the activity of TrioD1 requires the microtubule network and relocalizes RhoG at the active sites of the plasma membrane. Using a classical in vitro exchange assay, TrioD1 displays a higher GEF activity on RhoG than on Rac1. In fibroblasts, expression of dominant negative RhoG mutants totally abolished TrioD1 signaling, whereas dominant negative Rac1 and Cdc42Hs only led to partial and complementary inhibitions. Finally, expression of a Rho Binding Domain that specifically binds RhoG(GTP) led to the complete abolition of TrioD1 signaling, which strongly supports Rac1 not being activated by TrioD1 in vivo. These data demonstrate that Trio controls a signaling cascade that activates RhoG, which in turn activates Rac1 and Cdc42Hs.


Internal fixation for osteomyelitis of cervical spine: the issue of persistence of culture positive infection around the implants.
We describe the management of osteomyelitis of the cervical spine, utilizing internal fixation with subsequent removal and culture of the implants. Four out of five patients had evidence of bacterial colonisation in close proximity to the internal fixation device. Five consecutive patients (all female, ranging in age from 50 to 74 yrs) presenting with unstable cervical osteomyelitis were treated by surgical decompression, primary internal fixation followed by three months of intravenous antibiotics. The internal fixation was removed in 4 out of 5 cases within a year of stopping the intravenous regime. The remaining patient was deemed medically unfit for further operation. Multiple specimens from the screw sites were taken at the time of metal removal. A final course of oral antibiotics was prescribed based on the results of these specimens. Four patients, who had removal of the implants, had positive cultures growing different bacteria from the primary infection, at the time of removal of the implant. None of the patients developed instability after removal of the implant. Asymptomatic bacterial colonisation of a metallic implant has profound management implications. We recommend long-term oral antibiotic regimes after insertion of internal fixation devices in the face of infection and eventual removal of these implants and microbiological re-sampling.


Does less autonomy erode women's health? Yes. No. Maybe.
Understanding the determinants of health is a central objective of human biology and related fields. Female autonomy is hypothesized to be an important determinant of women's health as well as demographic outcomes. The literature relating women's health to their everyday autonomy has produced conflicting results, and this may be due in part to the application of different measures of autonomy and different measures of health. Using secondary data from a large nationally representative study, this study examines the relationship between multiple measures of female autonomy and three measures of wellbeing among women living in Uzbekistan (n = 5,396). The multivariate results show that women's autonomy related to freedom of movement is associated with lower levels of depression symptomatology and lower systolic blood pressure. Respondents who assert that women should have control over their bodies also had lower odds of high depression symptoms and lower diastolic blood pressure. In contrast, women with greater decision-making autonomy were more likely to be classified as having high depressive symptomatology and higher diastolic blood pressure. Building on recent work, we suggest that these associations might reflect varying levels of agreement between men and women, and we provide some limited evidence to support this. This study stands as a theoretical and methodological cautionary note by suggesting that the relationship between autonomy and health is complex. Further, if differences in gender agreement underlie differences in the predictive accuracy of autonomy scales, then human biology researchers will need to begin collecting identical data from men and women.


Coexistence of esophageal superficial carcinoma and multiple leiomyomas: A case report.
Leiomyomas are the most common benign tumors of the esophagus. They usually occur as a single lesion or as two or three nodules. Only two cases of esophageal multiple leiomyomas comprising more than 10 nodules have been reported previously. Moreover, there have been few reports of esophageal squamous cell carcinoma overlying submucosal tumors. We describe a 71-year-old man who was diagnosed as having a superficial esophageal cancer coexisting with two or three leiomyoma nodules. During surgery, 10 or more nodules that had not been evident preoperatively were palpable in the submucosal and muscular layers throughout the esophagus. As intramural metastasis of the esophageal cancer was suspected, we considered additional lymphadenectomy, but had to rule out this option because of the patient's severe anoxemia. Microscopic examination revealed that all the nodules were leiomyomas (20 lesions, up to 3 cm in diameter), and that invasion of the carcinoma cells was limited to the submucosal layer overlying a relatively large leiomyoma. This is the first report of superficial esophageal cancer coexisting with numerous solitary leiomyomas. Multiple minute leiomyomas are often misdiagnosed as intramural metastasis, and a leiomyoma at the base of a carcinoma lesion can also be misdiagnosed as tumor invasion. The present case shows that accurate diagnosis is required for the management of patients with coexisting superficial esophageal cancer and multiple leiomyomas.


Arabidopsis replacement histone variant H3.3 occupies promoters of regulated genes.
Histone variants establish structural and functional diversity of chromatin by affecting nucleosome stability and histone-protein interactions. H3.3 is an H3 histone variant that is incorporated into chromatin outside of S-phase in various eukaryotes. In animals, H3.3 is associated with active transcription and possibly maintenance of transcriptional memory. Plant H3 variants, which evolved independently of their animal counterparts, are much less well understood. We profile the H3.3 distribution in Arabidopsis at mono-nucleosomal resolution using native chromatin immunoprecipitation. This results in the precise mapping of H3.3-containing nucleosomes, which are not only enriched in gene bodies as previously reported, but also at a subset of promoter regions and downstream of the 3' ends of active genes. While H3.3 presence within transcribed regions is strongly associated with transcriptional activity, H3.3 at promoters is often independent of transcription. In particular, promoters with GA motifs carry H3.3 regardless of the gene expression levels. H3.3 on promoters of inactive genes is associated with H3K27me3 at gene bodies. In addition, H3.3-enriched plant promoters often contain RNA Pol II considerably upstream of the transcriptional start site. H3.3 and RNA Pol II are found on active as well as on inactive promoters and are enriched at strongly regulated genes. In animals and plants, H3.3 organizes chromatin in transcribed regions and in promoters. The results suggest a function of H3.3 in transcriptional regulation and support a model that a single ancestral H3 evolved into H3 variants with similar sub-functionalization patterns in plants and animals.


Assessing Marijuana Use During Pregnancy.
Recreational and medicinal marijuana uses have become more prevalent in recent years. Women's health nurses are increasingly likely to encounter pregnant women who use or have used marijuana. Research has found that inadequate screening for substance use, inadequate knowledge about the effects of marijuana use, and punitive counseling are barriers to women's understanding of the importance of seeking treatment for substance use. Provision of evidence-based counseling, education, and resources support women and strengthen the patient-provider relationship. Women's health nurses can gain a basic understanding of the effects of perinatal marijuana use and create a plan for women who have positive screening results. This article reviews two recent studies that examine marijuana use during pregnancy and suggest interventions that women's health nurses can implement.


Welwitindolinone analogues that reverse P-glycoprotein-mediated multiple drug resistance.
Welwitindolinones are a family of novel alkaloids recently isolated from the blue-green alga Hapalosiphon welwitschii as a part of our effort to identify new compounds that overcome multiple drug resistance. The abilities of three structurally similar members of this family to interact with P-glycoprotein have been compared. Similarly to the effects of verapamil, N-methylwelwitindolinone C isothiocyanate (compound 1) attenuated the resistance of MCF-7/ADR cells to natural product anticancer drugs, including vinblastine, taxol, actinomycin D, daunomycin, and colchicine, without affecting the cytotoxicity of cisplatin. These effects of compound 1 were apparent at doses as low as 0.1 microM, indicating that it is considerably more potent than verapamil for reversal of resistance. Welwitindolinone C isothiocyanate (compound 3) demonstrated weaker reversing activity, whereas an analogue of compound 1 in which the isothiocyanate group is replaced by an isonitrile group (compound 2) was inactive. The accumulation of [3H]vinblastine in SK-VLB-1 cells was increased by compound 1 > compound 3 > verapamil >> compound 2. Interestingly, only compound 1 and verapamil enhanced [3H]taxol accumulation by these cells. Photoaffinity labeling of P-glycoprotein with [3H]azidopine in membranes from SK-VLB-1 cells was inhibited by compounds 1 and 3, but not by compound 2. Therefore, the differences in the size and/or the electronegativity of the isothiocyanate and isonitrile moieties appear to dramatically affect the abilities of the compounds to interact with P-glycoprotein.


Accelerators: Sparking Innovation and Transdisciplinary Team Science in Disparities Research.
Development and implementation of effective, sustainable, and scalable interventions that advance equity could be propelled by innovative and inclusive partnerships. Readied catalytic frameworks that foster communication, collaboration, a shared vision, and transformative translational research across scientific and non-scientific divides are needed to foster rapid generation of novel solutions to address and ultimately eliminate disparities. To achieve this, we transformed and expanded a community-academic board into a translational science board with members from public, academic and private sectors. Rooted in team science, diverse board experts formed topic-specific "accelerators", tasked with collaborating to rapidly generate new ideas, questions, approaches, and projects comprising patients, advocates, clinicians, researchers, funders, public health and industry leaders. We began with four accelerators-digital health, big data, genomics and environmental health-and were rapidly able to respond to funding opportunities, transform new ideas into clinical and community programs, generate new, accessible, actionable data, and more efficiently and effectively conduct research. This innovative model has the power to maximize research quality and efficiency, improve patient care and engagement, optimize data democratization and dissemination among target populations, contribute to policy, and lead to systems changes needed to address the root causes of disparities.


Translational medicine as a permanent glue and force of clinical medicine and public health: perspectives (1) from 2012 Sino-American symposium on clinical and translational medicine.
Health systems globally face challenges and opportunities in balancing quality, access, and cost, where clinical and translational medicine (CTM) should play more important and powerful roles in the identification, development and validation of solutions and strategies. Strategic collaboration can gather global strengths and resources and improve health systems, care delivery, regulations and policies. CTM-driven innovation and development has the potential to achieve step-change improvements across three dimensions. Thus, we have the reasons to believe that CTM will play even more roles in the development of new diagnostics, therapies, healthcare, and policies and SAS-CTM will become more and more important platform to obtain the latest development in CTM internationally and explore new opportunities in the international collaborations.


The influence of food on postural hypotension in three groups with chronic autonomic failure--clinical and therapeutic implications.
The effect of a balanced liquid meal on supine and postural blood pressure (BP) responses was investigated in three groups of patients with chronic autonomic failure; 10 with associated neurological impairment (multiple system atrophy (MSA), Shy-Drager syndrome) and seven without (of which five had pure autonomic failure (PAF); and two had a deficiency of the enzyme dopamine beta hydroxylase, DBH-deficiency). All had marked postural hypotension. Subjects with normal autonomic function were also studied. In MSA and PAF food lowered supine BP substantially, with a more rapid and greater fall in PAF. After food, the levels of BP reached were considerably lower because of the reduced supine BP and many had to be returned to the horizontal position earlier than before. Ingestion of a similar volume of water alone had no effect in MSA or PAF. In DBH deficiency, food had variable but minimal effects on BP while supine and during head-up tilt. In subjects with normal autonomic function food did not affect BP. The BP responses to food thus varied in the three groups with chronic autonomic failure. The influence of food on both supine and postural BP therefore should be considered in the clinical and laboratory assessment of autonomic dysfunction and in relation to therapeutic approaches, designed to alleviate postural hypotension.


Integration of genome-level data to allow identification of subtype-specific vulnerability genes as novel therapeutic targets.
The identification of cancer-specific vulnerability genes is one of the most promising approaches for developing more effective and less toxic cancer treatments. Cancer genomes exhibit thousands of changes in DNA methylation and gene expression, with the vast majority likely to be passenger changes. We hypothesised that, through integration of genome-wide DNA methylation/expression data, we could exploit this inherent variability to identify cancer subtype-specific vulnerability genes that would represent novel therapeutic targets that could allow cancer-specific cell killing. We developed a bioinformatics pipeline integrating genome-wide DNA methylation/gene expression data to identify candidate subtype-specific vulnerability partner genes for the genetic drivers of individual genetic/molecular subtypes. Using acute lymphoblastic leukaemia as an initial model, 21 candidate subtype-specific vulnerability genes were identified across the five common genetic subtypes, with at least one per subtype. To confirm the approach was applicable across cancer types, we also assessed medulloblastoma, identifying 15 candidate subtype-specific vulnerability genes across three of four established subtypes. Almost all identified genes had not previously been implicated in these diseases. Functional analysis of seven candidate subtype-specific vulnerability genes across the two tumour types confirmed that siRNA-mediated knockdown induced significant inhibition of proliferation/induction of apoptosis, which was specific to the cancer subtype in which the gene was predicted to be specifically lethal. Thus, we present a novel approach that integrates genome-wide DNA methylation/expression data to identify cancer subtype-specific vulnerability genes as novel therapeutic targets. We demonstrate this approach is applicable to multiple cancer types and identifies true functional subtype-specific vulnerability genes with high efficiency.


Identification of the DNA region responsible for sulfur-oxidizing ability of Thiosphaera pantotropha.
For the identification of the DNA region responsible for the sulfur-oxidizing ability (Sox) of Thiosphaera pantotropha, we used previously isolated Tn5-mob insertional Sox- mutants. For seven mutants, the Tn5-mob insertion was localized on the chromosome rather than on the megaplasmids pHG41 or pHG42 by using the Tn5-mob-harboring vehicle pSUP5011 as probe. The specific insertion of Tn5-mob into a sox gene was determined for one Sox- mutant, strain TP19. An 18-kb EcoRI fragment was cloned in Escherichia coli by using the mobilizable plasmid pSUP202 as vector and the kanamycin resistance gene of Tn5 as marker. Conjugal transfer of the resulting hybrid plasmid, pKS3-13, to the wild type resulted in two phenotypically different groups of recombinants. Ninety-five percent of the recombinants were Sox+, kanamycin resistant, and tetracycline resistant; 5% were homogenote recombinants exhibiting the Sox-, kanamycin-resistant, tetracycline-sensitive phenotype, and these indicated the specific insertion. To isolate the respective wild-type sox gene, total DNA from a heterogenote recombinant was partially restricted with EcoRI, religated, and transformed in E. coli. Transformants carrying a pSUP202-derived hybrid plasmid with the intact sox gene were identified by screening for a tetracycline-resistant, kanamycin-sensitive, and chloramphenicol-sensitive phenotype and by complementation of the Sox- mutant TP19. A plasmid of this type, pEG12, contained an insert of 13 kb which gave a positive signal in Southern hybridization with the homologous probe of pKS3-13. pEG12 was used to determine the DNA homology of the sulfur-oxidizing enzyme systems of other thiobacteria. Strong hybridization signals were obtained with total DNA of the neutrophilic sulfur-oxidizing bacteria Paracoccus denitrificans, Thiobacillus versutus, and Rhodobacter capsulatus. No hybridization signal was obtained with DNA of other neutrophilic or acidophilic thiobacteria examined.


Vesicle trafficking and RNA transfer add complexity and connectivity to cell-cell communication.
Cell-cell communication, either in direct proximity or at a distance, generally occurs by receptor-ligand engagement and subsequent activation of downstream intracellular signaling cascades. This conventional, largely protein-based, model has long been considered necessary and sufficient to explain coordinate tissue and organismal function. Intriguing recent work indicates that many cells can also transfer RNA directly via cell-cell trafficking of nanometer-sized, lipid-bilayer vesicles. The distinct biogenesis pathways that give rise to the different vesicle types described to date are just beginning to be elucidated. Notwithstanding their diverse origin, all types of vesicles seem to contain a broad, cell-specific, nonrandom representation of cellular protein and RNA species. The cell-cell trafficking of coding and small noncoding RNAs in particular constitutes a new paradigm for the direct phenotypic modulation of cells in the local microenvironment and in distal organs. Here, we review the current understanding of RNA vesicle trafficking and its emerging role in cell-cell signaling.


Hydrogel-forming microneedles enhance transdermal delivery of metformin hydrochloride.
We investigated, for the first time, the potential for a hydrogel-forming microneedle (MN) patch to deliver the high-dose drug metformin HCl transdermally in a sustained manner. This may minimize some gastrointestinal side effects and small intestine absorption variations associated with oral delivery. Patches (two layers) were assembled from a lyophilised drug reservoir layer, with the MN layer made from aqueous blend of 20% w/w poly (methylvinylether-co-maleic acid) crosslinked by esterification with 7.5% w/w poly (ethylene glycol) 10,000 Da. >90% of metformin was recovered from homogeneous drug reservoirs. Drug reservoir dissolution time in PBS (pH 7.4) was <10 min. MN penetrated a validated skin model Parafilm® M consistently. Permeation of metformin HCl across dermatomed neonatal porcine skin in vitro was enhanced by using MN. The combined MN and metformin HCl reservoir patch (containing 75 mg or 50 mg metformin HCl, respectively) delivered 9.71 ± 2.22 mg and 10.04 ± 1.92 mg at 6 h, respectively, and 28.15 ± 2.37 mg and 23.25 ± 3.58 mg at 24 h, respectively.In comparison, 0.34 ± 0.39 mg and 0.85 ± 0.68 mg was delivered at 6 h, respectively, and 0.39 ± 0.39 mg and 1.01 ± 0.84 mg was delivered at 24 h, respectively, from a control set-up employing only the drug reservoirs. In vivo, metformin HCl was detected in rat plasma at 1 h post MN application at a concentration of 0.62 ± 0.51 μg/mL, increasing to 3.76 ± 2.58 μg/ml at 3 h. A maximal concentration of 3.77 ± 2.09 μg/ml was achieved at 24 h. Css was 3.2 μg/mL. Metformin transdermal bioavailability using MNs was estimated as 30%.Hydrogel-forming MN are a promising technology that has demonstrated successful transdermal delivery of metformin HCl. Potential clearly exists for administration of other high-dose drugs using this system.


Glucose transport by isolated plasma membranes of the bovine blood-brain barrier.
Luminal and abluminal endothelial plasma membrane vesicles were isolated from bovine cerebral microvessels, the site of the blood-brain barrier. Glucose transport across each membrane was measured using a rapid-filtration technique. Glucose transport into luminal vesicles occurred by a stereospecific energy-independent transporter [Michaelis-Menten constant (K(m)) = 10.3 +/- 2.8 (SE) mM and maximal velocity (Vmax) = 8.6 +/- 2.0 nmol.mg protein(-1).min-1]. Kinetic analysis of abluminal vesicles also showed a transport system with characteristics similar to the luminal transporter (K(m) = 12.5 +/- 2.3 mM and Vmax = 10.0 +/- 1.0 nmol.mg protein-1.min-1). These functional, facilitative glucose transporters were symmetrically distributed between the luminal and abluminal membrane domains, providing a mechanism for glucose movement between blood and brain. The studies also revealed a Na-dependent transporter on the abluminal membrane with a higher affinity and lower capacity than the facilitative transporters (K(m) = 130 +/- 20 microM and Vmax = 1.59 +/- 0.44 nmol.mg protein-1.min-1. The abluminal Na-dependent glucose transporter is in a position to transport glucose from the brain extracellular fluid into the endothelial cells of the blood-brain barrier. The functional significance of its presence there remains to be determined.


Serotonergic mechanism of the relieving effect of bee venom acupuncture on oxaliplatin-induced neuropathic cold allodynia in rats.
Oxaliplatin, an important chemotherapy drug for advanced colorectal cancer, often induces peripheral neuropathy, especially cold allodynia. Our previous study showed that bee venom acupuncture (BVA), which has been traditionally used in Korea to treat various pain symptoms, potently relieves oxaliplatin-induced cold allodynia in rats. However, the mechanism for this anti-allodynic effect of BVA remains poorly understood. We investigated whether and how the central serotonergic system, a well-known pathway for acupuncture analgesia, mediates the relieving effect of BVA on cold allodynia in oxaliplatin-injected rats. The behavioral signs of cold allodynia in Sprague-Dawley (SD) rats were induced by a single injection of oxaliplatin (6 mg/kg, i.p.). Before and after BVA treatment, the cold allodynia signs were evaluated by immersing the rat's tail into cold water (4°C) and measuring the withdrawal latency. For BVA treatment, a diluted BV (0.25 mg/kg) was subcutaneously administered into Yaoyangguan (GV3) acupoint, which is located between the spinous processes of the fourth and the fifth lumbar vertebra. Serotonin was depleted by a daily injection of DL-p-chlorophenylalanine (PCPA, 150 mg/kg, i.p.) for 3 days. The amount of serotonin in the spinal cord was measured by ELISA. Serotonergic receptor antagonists were administered intraperitoneally or intrathecally before BVA treatment. The serotonin levels in the spinal cord were significantly increased by BVA treatment and such increase was significantly reduced by PCPA. This PCPA pretreatment abolished the relieving effect of BVA on oxaliplatin-induced cold allodynia. Either of methysergide (mixed 5-HT1/5-HT2 receptor antagonist, 1 mg/kg, i.p.) or MDL-72222 (5-HT3 receptor antagonist, 1 mg/kg, i.p) blocked the anti-allodynic effect of BVA. Further, an intrathecal injection of MDL-72222 (12 μg) completely blocked the BVA-induced anti-allodynic action, whereas NAN-190 (5-HT1A receptor antagonist, 15 μg, i.t.) or ketanserin (5-HT2A receptor antagonist, 30 μg, i.t.) did not. These results suggest that BVA treatment alleviates oxaliplatin-induced acute cold allodynia in rats via activation of the serotonergic system, especially spinal 5-HT3 receptors. Thus, our findings may provide a clinically useful evidence for the application of BVA as an alternative therapeutic option for the management of peripheral neuropathy, a dose-limiting side effect that occurs after an administration of oxaliplatin.


Developing a statewide emergency medical services database linked to hospital outcomes: a feasibility study.
Statewide emergency medical services (EMS) data linked to outcomes are critical for promoting high-quality emergency care; however, many states do not have such a resource. To demonstrate the feasibility of creating such a statewide database using a one-month pilot sample. This was a prospective cohort study of all EMS patient encounters throughout Oregon during May 2008. Eighty-three National EMS Information System (NEMSIS) variables were obtained from EMS agencies via electronic or paper charts. We reformatted raw data, mapped NEMSIS fields, entered hard-copy records, and uploaded data files to a statewide electronic medical records platform. Records from transport and nontransport (first-responder) agencies caring for the same patients were matched using probabilistic linkage, then linked to three statewide outcome databases (Oregon Hospital Discharge Database [OHDD], Oregon Trauma Registry [OTR], and Oregon Department of Transportation [ODOT] Crash File) using similar methodology. We estimated population-adjusted case ascertainment by county and used descriptive statistics to characterize the process. During the one-month period, we collected 27,474 EMS records in 36 (100%) counties from 106 (77%) licensed transport agencies and 10 nontransport agencies, representing 20,673 persons. There were 3,302 admission record matches, 285 trauma registry matches, and 392 crash record matches. Overall, 3,979 hospital outcomes were matched to EMS records for 80 (75%) transport and six (60%) first-responder agencies. Median per-agency match rates were 16.3% for OHDD (interquartile range [IQR] 8.3-22.2%, range 0-56.5%), 0.9% for OTR (IQR 0-2.5%, range 0-60.0%), and 1.6% for ODOT (IQR 0-3.5%, range 0-23.1%). Developing a statewide EMS database linked to hospital outcomes is feasible. The processes used in this study and match rate estimates may provide a template for other states to follow, enhancing opportunities for outcomes-based EMS research and EMS quality assurance efforts.


Comparative genomic hybridization on microarray (a-CGH) in constitutional and acquired mosaicism may detect as low as 8% abnormal cells.
The results of cytogenetic investigations on unbalanced chromosome anomalies, both constitutional and acquired, were largely improved by comparative genomic hybridization on microarray (a-CGH), but in mosaicism the ability of a-CGH to reliably detect imbalances is not yet well established. This problem of sensitivity is even more relevant in acquired mosaicism in neoplastic diseases, where cells carrying acquired imbalances coexist with normal cells, in particular when the proportion of abnormal cells may be low.We constructed a synthetic mosaicism by mixing the DNA of three patients carrying altogether seven chromosome imbalances with normal sex-matched DNA. Dilutions were prepared mimicking 5%, 6%, 7%, 8%, 10% and 15% levels of mosaicism. Oligomer-based a-CGH (244 K whole-genome system) was applied on the patients' DNA and customized slides designed around the regions of imbalance were used for the synthetic mosaics. The a-CGH on the synthetic mosaics proved to be able to detect as low as 8% abnormal cells in the tissue examined. Although in our experiment some regions of imbalances escaped to be revealed at this level, and were detected only at 10-15% level, it should be remarked that these ones were the smallest analyzed, and that the imbalances recurrent as clonal anomalies in cancer and leukaemia are similar in size to those revealed at 8% level.


Extremely low-frequency electromagnetic fields cause DNA strand breaks in normal cells.
Extremely low frequency electromagnetic fields aren't considered as a real carcinogenic agent despite the fact that some studies have showed impairment of the DNA integrity in different cells lines. The aim of this study was evaluation of the late effects of a 100 Hz and 5.6 mT electromagnetic field, applied continuously or discontinuously, on the DNA integrity of Vero cells assessed by alkaline Comet assay and by cell cycle analysis. Normal Vero cells were exposed to extremely low frequency electromagnetic fields (100 Hz, 5.6 mT) for 45 minutes. The Comet assay and cell cycle analysis were performed 48 hours after the treatment. Exposed samples presented an increase of the number of cells with high damaged DNA as compared with non-exposed cells. Quantitative evaluation of the comet assay showed a significantly (<0.001) increase of the tail lengths, of the quantity of DNA in tail and of Olive tail moments, respectively. Cell cycle analysis showed an increase of the frequency of the cells in S phase, proving the occurrence of single strand breaks. The most probable mechanism of induction of the registered effects is the production of different types of reactive oxygen species. The analysis of the registered comet indices and of cell cycle showed that extremely low frequency electromagnetic field of 100 Hz and 5.6 mT had a genotoxic impact on Vero cells.


Production of calcium-mobilizing metabolites by a novel member of the ADP-ribosyl cyclase family expressed in Schistosoma mansoni.
ADP-ribosyl cyclases are structurally conserved enzymes that are best known for catalyzing the production of the calcium-mobilizing metabolite, cyclic adenosine diphosphate ribose (cADPR), from nicotinamide adenine dinucleotide (NAD(+)). However, these enzymes also produce adenosine diphosphate ribose (ADPR) and nicotinic acid adenine dinucleotide phosphate (NAADP(+)), both of which have been shown to modulate calcium mobilization in vitro. We have now characterized a new member of the cyclase family from Schistosoma mansoni, a member of the Platyhelminthes phylum. We show that the novel NAD(P)(+) catabolizing enzyme (NACE) expressed by schistosomes is structurally most closely related to the cyclases cloned from Aplysia but also shows significant homology with the mammalian cyclases, CD38 and CD157. NACE expression is developmentally regulated in schistosomes, and the GPI-anchored protein is localized to the outer tegument of the adult schistosome. Importantly, NACE, like all members of the cyclase family, is a multifunctional enzyme and catalyzes NAD(+) glycohydrolase and base-exchange reactions to produce ADPR and NAADP(+). However, despite being competent to generate a cyclic product from NGD(+), a nonphysiologic surrogate substrate, NACE is so far the only enzyme in the cyclase family that is unable to produce significant amounts of cADPR (<0.02% of reaction products) using NAD(+) as the substrate. This suggests that the other calcium-mobilizing metabolites produced by NACE may be more important for calcium signaling in schistosomes. Alternatively, the function of NACE may be to catabolize extracellular NAD(+) to prevent its use by host enzymes that utilize this source of NAD(+) to facilitate immune responses.


Global kinomic and phospho-proteomic analyses of the human malaria parasite Plasmodium falciparum.
The role of protein phosphorylation in the life cycle of malaria parasites is slowly emerging. Here we combine global phospho-proteomic analysis with kinome-wide reverse genetics to assess the importance of protein phosphorylation in Plasmodium falciparum asexual proliferation. We identify 1177 phosphorylation sites on 650 parasite proteins that are involved in a wide range of general cellular activities such as DNA synthesis, transcription and metabolism as well as key parasite processes such as invasion and cyto-adherence. Several parasite protein kinases are themselves phosphorylated on putative regulatory residues, including tyrosines in the activation loop of PfGSK3 and PfCLK3; we show that phosphorylation of PfCLK3 Y526 is essential for full kinase activity. A kinome-wide reverse genetics strategy identified 36 parasite kinases as likely essential for erythrocytic schizogony. These studies not only reveal processes that are regulated by protein phosphorylation, but also define potential anti-malarial drug targets within the parasite kinome.


Psychiatric diagnosis and antiretroviral adherence among adolescent Medicaid beneficiaries diagnosed with human immunodeficiency virus/acquired immunodeficiency syndrome.
Research on adults with human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) has suggested that psychiatric and substance abuse comorbidities are prevalent in this population, and that these may sometimes be associated with use of antiretroviral therapy (ART) and adherence. For adolescents with HIV/AIDS, much less is known about patterns of mental health comorbidity, and even fewer data are available that compare them to socioeconomically comparable youth without HIV/AIDS. Using medical and pharmacy data from 1999 to 2000 Medicaid claims (Medicaid Analytic Extract) from 4 states for beneficiaries aged 12 to 17 years, we identified 833 youth under care for HIV/AIDS meeting study criteria within the HIV/AIDS group, receipt of ART was less likely for youth who had diagnoses of substance abuse, conduct disorders, or emotional disorders than for others. Once ART was initiated, adherence did not significantly differ between adolescents living with a psychiatric condition, and those who were not, with the exception of an association between conduct disorder and lower adherence. Among those with HIV/AIDS, ART use and adherence were more common among youth with higher rates of service use, regardless of psychiatric status. Associations between race and adherence varied by gender: compared with their white counterparts, minority girls had lower, and minority boys had higher adherence.


Society for Endocrinology UK Guidance on the initial evaluation of a suspected difference or disorder of sex development (Revised 2021).
It is paramount that any child or adolescent with a suspected difference or disorder of sex development (DSD) is assessed by an experienced clinician with adequate knowledge about the range of conditions associated with DSD and is discussed with the regional DSD service. In most cases, the paediatric endocrinologist within this service acts as the first point of contact but involvement of the regional multidisciplinary service will also ensure prompt access to specialist psychology and nursing care. The underlying pathophysiology of DSD and the process of delineating this should be discussed with the parents and affected young person with all diagnostic tests undertaken in a timely fashion. Finally, for rare conditions such as these, it is imperative that clinical experience is shared through national and international clinical and research collaborations.


Five cases of forensic short tandem repeat DNA typing.
In medicolegal samples DNA is often broken into fragments. In many cases, only the amplification of short tandem repeated DNA stretches (STRs), which are located in noncoding regions, allows DNA typing of such degraded materials. To demonstrate the high diversity of biological materials which forensic biologists have to deal with, and to outline the success rates and limits of the method, we describe five cases (minute amount of tissue on barrel, tissue in decay, tumor tissue, sperm after multiple rape, stored urine samples) in which forensic DNA typing was successfully performed by use of the short tandem repeats HUMDHFRP2, HUMD8S306, HUMCD4, HUMF13A1, HUMTH01, HUMVWA, and HUMFES.


Effects of indium exposure on respiratory symptoms: a retrospective cohort study in Japanese workers using health checkup data.
Indium compounds are known health hazards for lung cancer and interstitial pneumonia. Furthermore, they are related to emphysema, alveolar proteinosis, and cholesterol granuloma. In Japan, laws were revised in 2013 to tighten regulations on indium exposure in workplaces. However, its impact on the health of workers who handle indium has not been evaluated. This study aimed to investigate whether subjective respiratory symptoms in these workers have reduced after the 2013 amendment in the regulations. The subjects were workers from certain areas of Japan who had undergone health checkups between January 1, 2013, and June 30, 2015. Indium-handling and non-handling workers were categorized into the exposed and less-exposed groups, respectively. Based on the findings of health checkups during this period, the hazard ratio of subjective respiratory symptoms (cough, sputum production, shortness of breath, and palpitation) and its 95% confidence intervals (CIs) were calculated with the less-exposed group as the reference. The Prentice-Williams-Peterson model was used for calculation, and a model that adjusted for coarse analysis and potential confounding factors was adopted. Overall, 2,561 workers (from 22 companies) who underwent 6,033 health checkups were included. The total person-years were 2,562.8 years, and 162 outcome events occurred. The hazard ratios of the exposed group were 1.65 (95% CI [1.14-2.39]: p = 0.008) and 1.61 (95% CI [1.04-2.50]: p = 0.032) in the crude and adjusted models, respectively. Indium-handling workers had a high hazard of the subjective respiratory symptoms than non-indium -handling workers despite stricter regulations on indium exposure in workplaces. This indicates the need for further changes to the legislation to protect the health of workers exposed to harmful substances in workplaces. Further studies including larger diverse cohorts are needed to validate our findings.


Use of screening and preventive services among women with disabilities.
Roughly 54 million Americans have some disability; at older ages, women are more likely to be disabled than men. Many people with disabilities today live virtually normal life spans, and therefore routine screening and preventive services are essential to their overall quality of care. We used the 1994-1995 National Health Interview Survey (NHIS), with Disability, Family Resources, and Healthy People 2000 supplements, to examine screening and preventive service use for adult women with disabilities living in the community--about 18.4% of women (estimated 18.28 million). Disability was associated with higher age-adjusted rates of: poverty; living alone; low education; inability to work; obesity; and being frequently depressed or anxious. Disabled women generally reported screening and preventive services at rates comparable to all women. Women with major lower extremity mobility difficulties had much lower adjusted odds of Papanicolaou smears (odds ratio, 0.6; 95% confidence interval, 0.4-0.9), mammograms (odds ratio, 0.7; 95% confidence interval, 0.5-0.9), and smoking queries (odds ratio, 0.6; 95% confidence interval, 0.5-0.8). Various approaches exist to improve access for disabled women to health care services.


Enhanced reactive lysis of paroxysmal nocturnal hemoglobinuria erythrocytes. Studies on C9 binding and incorporation into high molecular weight complexes.
As part of a broader analysis of the mechanism(s) by which the most sensitive (type III) paroxysmal nocturnal hemoglobinuria (PNH) erythrocytes are excessively sensitive to reactive lysis by isolated C5b6, C7, C8, and C9, we have compared type III PNH (PNH-III) and normal human E in respect to both total specific binding of 125I-C9 and the proportion of cell-bound C9 appearing in high molecular weight (HMW) complexes. In a previous report, we found that after exposure to purified C5b6 and 125I-C7, specific C7 binding and, by implication, EC5b-7 formation were equal for PNH-III E and normal E. In the present study, C8-dependent binding of 125I-C9 to PNH-III EC5b-7 and normal EC5b-7 was also similar, although lysis of the PNH-III E was up to five times greater; that is, PNH-III E required fewer bound C9 molecules to produce an effective lytic site than did normal E. To quantify radioactivity in monomeric and HMW forms of membrane-bound C9, lysed and unlysed E were subjected to low ionic strength buffers to convert all E to ghosts. These ghosts were solubilized in 0.1 or 2% SDS (without reduction) and electrophoresed on 2.4-11% polyacrylamide gradient gels followed by autoradiography and densitometric scanning. With 0.1% SDS, broad, heterodisperse zones of HMW C9 were recovered from both PNH and normal ghosts; the amounts of C9 incorporated into the HMW complexes were similar for PNH-III E and normal E. In selected experiments, 125I-C7 could be shown in these same HMW bands. When membranes were solubilized in 2% SDS, the overall proportion of HMW C9 complexes compared with dimer and monomer C9 was reduced on both PNH and normal membranes. In many, but not all experiments, more of the highest mol wt C9 complexes were detected from PNH-III E membranes solubilized in 2% SDS than from normal or PNH-II E membranes similarly treated. When antibody-sensitized E were lysed by purified C1-C9, PNH-III EA bound far more C9 than did normal EA, and both lysis and C9 incorporation into HMW complexes were markedly and proportionately increased over normal; however, lytic efficiency of 125I-C9 bound to PNH EA was equal to or less than that bound to normal EA.(ABSTRACT TRUNCATED AT 250 WORDS)


A physiological controller for turbodynamic ventricular assist devices based on a measurement of the left ventricular volume.
The current article presents a novel physiological control algorithm for ventricular assist devices (VADs), which is inspired by the preload recruitable stroke work. This controller adapts the hydraulic power output of the VAD to the end-diastolic volume of the left ventricle. We tested this controller on a hybrid mock circulation where the left ventricular volume (LVV) is known, i.e., the problem of measuring the LVV is not addressed in the current article. Experiments were conducted to compare the response of the controller with the physiological and with the pathological circulation, with and without VAD support. A sensitivity analysis was performed to analyze the influence of the controller parameters and the influence of the quality of the LVV signal on the performance of the control algorithm. The results show that the controller induces a response similar to the physiological circulation and effectively prevents over- and underpumping, i.e., ventricular suction and backflow from the aorta to the left ventricle, respectively. The same results are obtained in the case of a disturbed LVV signal. The results presented in the current article motivate the development of a robust, long-term stable sensor to measure the LVV.


A review of penile elongation surgery.
Penile elongation surgery is less commonly performed in the public sector, but involves a collaborative approach between urology and plastic surgery. Congenital and acquired micropenis are the classic surgical indications for penile elongation surgery. The goal of intervention in these patients is to restore a functional penis size in order to allow normal standing micturition, enable satisfying sexual intercourse and improve patient quality of life. Many men seeking elongation actually have normal length penises, but perceive themselves to be small, a psychologic condition termed 'penile dysmorphophobia'. This paper will review the anatomy and embryology of congenital micropenis and discuss both conservative and surgical management options for men seeking penile elongation therapy.


Time-shift technique for simultaneous measurement of size, velocity, and relative refractive index of transparent droplets or particles in a flow.
The time-shift technique, also known as the pulsed-displacement technique, is revisited as a method to measure size, velocity, and relative refractive index of spherical, transparent particles. Building on the basic measurement principle, we introduce several new innovations, making the technique significantly more attractive for use outside of the laboratory. These innovations include two possibilities for velocity measurement, validation criteria for one- and two-detector arrangements, and approaches to achieve higher bandwidths, and, in particular, lower measurable sizes. We discuss numerous optical configuration examples to illustrate the flexibility of this technique to meet various application requirements.


Atrial fibrillation as a complication of dengue hemorrhagic fever: non-self-limiting manifestation.
While electrocardiogram (ECG) changes are common during viral dengue infection, atrial fibrillation (AF) is a very rare manifestation. It has previously been highlighted that cardiac complications during dengue infection are invariably transient and will spontaneously resolve following recovery from the illness. We present the case of a young patient with IgM- and IgG-positive dengue hemorrhagic fever complicated by AF. ECG revealed a structurally normal heart. The patient remained in AF despite resolution of the illness. Reversion to normal sinus rhythm was achieved after loading of oral amiodarone.


In situ localization of mitochondrial DNA replication in intact mammalian cells.
Nearly all of the known activities required for mitochondrial DNA (mtDNA) replication and expression are nuclear-encoded gene products, necessitating communication between these two physically distinct intracellular compartments. A significant amount of both general and specific biochemical information about mtDNA replication in mammalian cells has been known for almost two decades. Early studies achieved selective incorporation of the thymidine analog 5-Bromo-2-deoxy-Uridine (BrdU) into mtDNA of thymidine kinase-deficient (TK[-]) cells. We have revisited this approach from a cellular perspective to determine whether there exist spatiotemporal constraints on mtDNA replication. Laser-scanning confocal microscopy was used to selectively detect mtDNA synthesis in situ in cultured mammalian cells using an immunocytochemical double-labeling approach to visualize the incorporation of BrdU into mtDNA of dye-labeled mitochondria. In situ detection of BrdU-incorporated mtDNA was feasible after a minimum of 1-2 h treatment with BrdU, consistent with previous biochemical studies that determined the time required for completion of a round of mtDNA replication. Interestingly, the pattern of BrdU incorporation into the mtDNA of cultured mammalian cells consistently radiated outward from a perinuclear position, suggesting that mtDNA replication first occurs in the vicinity of nuclear-provided materials. Newly replicated mtDNA then appears to rapidly distribute throughout the dynamic cellular mitochondrial network.


NOTCH and DNA repair pathways are more frequently targeted by genomic alterations in inflammatory than in non-inflammatory breast cancers.
Inflammatory breast cancer (IBC) is the most pro-metastatic form of breast cancer. Better understanding of its pathophysiology and identification of actionable genetic alterations (AGAs) are crucial to improve systemic treatment. We aimed to define the DNA profiles of IBC vs noninflammatory breast cancer (non-IBC) clinical samples in terms of copy number alterations (CNAs), mutations, and AGAs. We applied targeted next-generation sequencing (tNGS) and array-comparative genomic hybridization (aCGH) to 57 IBC and 50 non-IBC samples and pooled these data with four public datasets profiled using NGS and aCGH, leading to a total of 101 IBC and 2351 non-IBC untreated primary tumors. The respective percentages of each molecular subtype [hormone receptor-positive (HR+)/HER2-, HER2+, and triple-negative] were 68%, 15%, and 17% in non-IBC vs 25%, 35%, and 40% in IBC. The comparisons were adjusted for both the molecular subtypes and the American Joint Committee on Cancer (AJCC) stage. The 10 most frequently altered genes in IBCs were TP53 (63%), HER2/ERBB2 (30%), MYC (27%), PIK3CA (21%), BRCA2 (14%), CCND1 (13%), GATA3 (13%), NOTCH1 (12%), FGFR1 (11%), and ARID1A (10%). The tumor mutational burden was higher in IBC than in non-IBC. We identified 96 genes with an alteration frequency (p < 5% and q < 20%) different between IBC and non-IBC, independently from the molecular subtypes and AJCC stage; 95 were more frequently altered in IBC, including TP53, genes involved in the DNA repair (BRCA2) and NOTCH pathways, and one (PIK3CA) was more frequently altered in non-IBC. Ninety-seven percent of IBCs displayed at least one AGA. This percentage was higher than in non-IBC (87%), notably for drugs targeting DNA repair, NOTCH signaling, and CDK4/6, whose pathways were more frequently altered (DNA repair) or activated (NOTCH and CDK4/6) in IBC than in non-IBC. The genomic landscape of IBC is different from that of non-IBC. Enriched AGAs in IBC may explain its aggressiveness and provide clinically relevant targets.


Action learning for health system governance: the reward and challenge of co-production.
Health policy and systems research (HPSR) is centrally concerned with people, their relationships and the actions and practices they can implement towards better health systems. These concerns suggest that HPS researchers must work in direct engagement with the practitioners and practice central to the inquiry, acknowledging their tacit knowledge and drawing it into generating new insights into health system functioning. Social science perspectives are of particular importance in this field because health policies and health systems are themselves social and political constructs. However, how can social science methodologies such as action research and narrative and appreciative enquiry enable such research, and how can methodologies from different disciplines be woven together to construct and make meaning of evidence for 'this' field? This article seeks to present 'methodological musings' on these points, to prompt wider discussion on the practice of HPSR. It draws on one long-term collaborative action learning research project being undertaken in Cape Town, South Africa. The District Innovation and Action Learning for Health System Development project is an action research partnership between two South African academic institutions and two health authorities focused, ultimately, on strengthening governance in primary health care.Drawing on this experience, the article considers three interrelated issues: The diversity and complexities of practitioner and research actors involved in co-producing HPSR; The nature of co-production and the importance of providing space to grapple across different systems of meaning;The character of evidence and data in co-production. There is much to be learnt from research traditions outside the health sector, but HPSR must work out its own practices--through collaboration and innovation among researchers and practitioners. In this article, we provide one set of experiences to prompt wider reflection and stimulate engagement on the practice of HPSR for people-centred health systems.


High diversity of non-sporulating moulds in respiratory specimens of immunocompromised patients: should all the species be reported when diagnosing invasive aspergillosis?
Non-sporulating moulds (NSMs) isolated from respiratory specimens are usually discarded without further testing although they may have pathogenic effects in immunocompromised patients. The objective of this study was to determine the identity and frequency of NSMs in patients with haematological malignancies. We analysed the mycological results of 251 consecutive respiratory samples from 104 haematology patients. Yeast and sporulating moulds were identified at the genus/species level according to their phenotypic features. NSMs were identified by internal transcribed spacer (ITS) sequencing. We detected 179 positive samples, of which 10.1% (18/179) were mixtures of moulds and 26.3% (47/179) were mixtures of moulds and yeast. We identified 142 moulds belonging to 11 different genera/species or groups, with Aspergillus fumigatus (n = 50), Penicillium spp. (n = 31) and NSM (n = 24) being the most frequently isolated species. Twenty-two NSMs were successfully sequenced: 18 were basidiomycetes and six were ascomycetes, corresponding to 16 different genera/species. NSMs were isolated with A. fumigatus in the same sample or in a subsequent sample in five patients with probable invasive aspergillosis. The conclusion is that the respiratory specimens of immunocompromised patients frequently contain very diverse mould species that may increase the virulence of pathogenic species. Reporting all mould species isolated when diagnosing invasive fungal infection could test this hypothesis.


Ignoring irrelevant stimuli in latent inhibition and Stroop paradigms: the effects of schizotypy and gender.
Latent inhibition (LI), poor evidence of learning following preexposure to a task-irrelevant stimulus, reflects the ability to ignore inconsequential events. Stroop interference represents a failure to inhibit processing of a task-irrelevant word when it is incongruent with the required naming of the word's print color. The apparent commonality between the two effects is in contradiction to the literature, which indicates that LI is affected by schizotypy and schizophrenia, and perhaps gender, while Stroop interference generated by the trial-to-trial procedure is unaltered by those variables. In the present experiment, low schizotypal healthy males, but not females, exhibited LI. The same groups did not differ on Stroop interference. The results are discussed in terms of different processing requirements for task-irrelevant stimuli that are an integral part of the task-relevant target stimulus (as in Stroop) or separated from it in space (as in LI).


Chip based common-path optical coherence tomography system with an on-chip microlens and multi-reference suppression algorithm.
We demonstrate an integrated optical probe including an on-chip microlens for a common-path swept-source optical coherence tomography system. This common-path design uses the end facet of the silicon oxynitride waveguide as the reference plane, thus eliminating the need of a space-consuming and dispersive on-chip loop reference arm, thereby obviating the need for dispersion compensation. The on-chip micro-ball lens eliminates the need of external optical elements for coupling the light between the chip and the sample. The use of this lens leads to a signal enhancement up to 37 dB compared to the chip without a lens. The light source, the common-path arm and the detector are connected by a symmetric Y junction having a wavelength independent splitting ratio (50/50) over a much larger bandwidth than can be obtained with a directional coupler. The signal-to-noise ratio of the system was measured to be 71 dB with 2.6 mW of power on a mirror sample at a distance of 0.3 mm from the waveguide end facet. Cross-sectional OCT images of a layered optical phantom sample are demonstrated with our system. A method, based on an extended Fourier-domain OCT model, for suppressing ghost images caused by additional parasitic reference planes is experimentally demonstrated.


A local space-time kriging approach applied to a national outpatient malaria dataset.
Increases in the availability of reliable health data are widely recognised as essential for efforts to strengthen health-care systems in resource-poor settings worldwide. Effective health-system planning requires comprehensive and up-to-date information on a range of health metrics and this requirement is generally addressed by a Health Management Information System (HMIS) that coordinates the routine collection of data at individual health facilities and their compilation into national databases. In many resource-poor settings, these systems are inadequate and national databases often contain only a small proportion of the expected records. In this paper we take an important health metric in Kenya (the proportion of outpatient treatments for malaria, MP) from the national HMIS database and predict the values of MP at facilities where monthly records are missing. The available MP data were densely distributed across a spatiotemporal domain and displayed second-order heterogeneity. We used three different kriging methodologies to make cross-validation predictions of MP in order to test the effect on prediction accuracy of (a) the extension of a spatial-only to a space-time prediction approach, and (b) the replacement of a globally-stationary with a locally-varying random function model. Space-time kriging was found to produce predictions with 98.4% less mean bias and 14.8% smaller mean imprecision than conventional spatial-only kriging. A modification of space-time kriging that allowed space-time variograms to be recalculated for every prediction location within a spatially-local neighbourhood resulted in a larger decrease in mean imprecision over ordinary kriging (18.3%) although mean bias was reduced less (87.5%).


Anatomical differences between CBS-corticobasal degeneration and CBS-Alzheimer's disease.
We compare patterns of gray matter loss on MRI in subjects presenting as corticobasal syndrome (CBS) with Alzheimer disease pathology (CBS-AD) to those presenting as CBS with corticobasal degeneration pathology (CBS-CBD). Voxel-based morphometry was used to compare patterns of gray matter loss in pathologically confirmed CBS-AD subjects (n = 5) and CBS-CBD subjects (n = 6) to a group of healthy controls (n = 20), and to each other. Atlas based parcellation using the automated anatomic labeling atlas was also utilized in a region-of-interest analysis to account for laterality. The CBS-AD subjects were younger at the time of scan when compared with CBS-CBD subjects (median: 60 years vs. 69; P = 0.04). After adjusting for age at time of MRI scan, the CBS-AD subjects showed loss in posterior frontal, temporal, and superior and inferior parietal lobes, while CBS-CBD showed more focal loss predominantly in the posterior frontal lobes when compared with controls. In both CBS-AD and CBS-CBD groups, there was basal ganglia volume loss, yet relative sparing of hippocampi. On direct comparisons between the two subject groups, CBS-AD showed greater loss in both temporal and inferior parietal cortices than CBS-CBD. No regions showed greater loss in the CBS-CBD group compared to the CBS-AD group. These findings persisted when laterality was taken into account. In subjects presenting with CBS, prominent temporoparietal, especially posterior temporal and inferior parietal, atrophy may be a clue to the presence of underlying AD pathology.


Some ADHD polymorphisms (in genes DAT1, DRD2, DRD3, DBH, 5-HTT) in case-control study of 100 subjects 6-10 age.
Pharmacological approach is the most effective way of treatment of ADHD and its early application prevents from the progress of secondary disorders. The study on present neurotransmitter systems in pathology of ADHD can be helpful in selecting appropriate drug, since there are used various substances with different mechanisms of functioning in treatment of the hyperkinetic syndrome. Within our study there were selected the genes of dopaminergic (DRD2, DRD3, DAT1), noradrenergic (DBH) and serotoninergic (5-HTT) systems. With the use of molecular-genetic methods based on association strategy "case-control" there were analysed genes including 11 polymorphisms. The presence of risk alleles was examined in comparison of the sample of 100 ADHD children to a control group of another 100 subjects, who were checked by child psychiatrists and examined with the Conners test in order to exclude eventual cases with ADHD symptoms. Our research suggests the association of some genes with ADHD. It could be concluded: 1) the risk of ADHD is significantly increased in the presence of one risk allele in genes DRD2 (O.R.=7,5), 5-HTT (O.R.=2,7) and DAT1 (O.R.=1,6). 2) The risk of ADHD is significantly increased at homozygotes for risk alleles in genes DRD2 (O.R.=54,8), 5-HTT (O.R.=6,7) and DAT1 (O.R.=6,6). For polymorphisms G444A and C1603T in DBH, which were detected by univariant analysis, haplotype analysis was performed and resulted in conclusion that: 3) the risk of ADHD is significantly increased in the presence of allele DBH +444A as well as in the presence of allele DBH +1603T (O.R.=15).


Behavioral problems in children with epilepsy in rural Kenya.
The aims of this study were to record behavioral problems in children with epilepsy (CWE), compare the prevalence with that reported among healthy children without epilepsy, and investigate the risk factors. A child behavioral questionnaire for parents comprising 15 items was administered to the main caregiver of 108 CWE and 108 controls matched for age in Kilifi, Kenya. CWE had a higher mean score for reported behavioral problems than controls (6.9 vs 4.9, t=4.7, P<0.001). CWE with active epilepsy also recorded more behavioral problems than those with inactive epilepsy (8.2 vs 6.2, t=-2.9, P=0.005). A significantly greater proportion of CWE (49% vs 26% of controls) were reported to have behavioral problems. Active epilepsy, cognitive impairment, and focal seizures were the most significant independent covariates of behavioral problems. Behavioral problems in African CWE are common and need to be taken into consideration in planning comprehensive clinical services in this region.


[Multivariate analyses using quantitative method II of annoyance due to tinnitus].
Factors affecting annoyance due to tinnitus were analyzed using quantitative method II. Ninety-one patients with unilateral tinnitus of single kind were studied. Major factors affecting annoyance due to tinnitus were found to be subjective loudness of tinnitus, age, degree of hearing loss, tinnitus loudness level in dB HL, pitch matched frequency, puerility and impurity of tinnitus tonality, and subjective tinnitus pitch. Increased annoyance due to tinnitus was associated with an age of 30-49 years, a tinnitus loudness level between 25 and 49 dB HL, a tinnitus loudness level more than 15dB SL, impurity of tinnitus tonality, larger subjective loudness, and high-pitched tones above 4000Hz. Decreased annoyance due to tinnitus was related to the smaller subjective loudness, loudness level below 25dB HL, low-pitched tones below 250Hz, and purity of tinnitus tonality. Our results suggest that these factors are responsible for individual differences in annoyance caused by tinnitus.


ALBA proteins are stage regulated during trypanosome development in the tsetse fly and participate in differentiation.
The protozoan parasite Trypanosoma brucei is responsible for sleeping sickness and alternates between mammal and tsetse fly hosts, where it has to adapt to different environments. We investigated the role of two members of the ALBA family, which encodes hypothetical RNA-binding proteins conserved in most eukaryotes. We show that ALBA3/4 proteins colocalize with the DHH1 RNA-binding protein and with a subset of poly(A+) RNA in stress granules upon starvation. Depletion of ALBA3/4 proteins by RNA interference in the cultured procyclic stage produces cell modifications mimicking several morphogenetic aspects of trypanosome differentiation that usually take place in the fly midgut. A combination of immunofluorescence data and videomicroscopy analysis of live trypanosomes expressing endogenously ALBA fused with fluorescent proteins revealed that ALBA3/4 are present throughout the development of the parasite in the tsetse fly, with the striking exception of the transition stages found in the proventriculus region. This involves migration of the nucleus toward the posterior end of the cell, a phenomenon that is perturbed upon forced expression of ALBA3 during the differentiation process, showing for the first time the involvement of an RNA-binding protein in trypanosome development in vivo.


Many-core algorithms for statistical phylogenetics.
Statistical phylogenetics is computationally intensive, resulting in considerable attention meted on techniques for parallelization. Codon-based models allow for independent rates of synonymous and replacement substitutions and have the potential to more adequately model the process of protein-coding sequence evolution with a resulting increase in phylogenetic accuracy. Unfortunately, due to the high number of codon states, computational burden has largely thwarted phylogenetic reconstruction under codon models, particularly at the genomic-scale. Here, we describe novel algorithms and methods for evaluating phylogenies under arbitrary molecular evolutionary models on graphics processing units (GPUs), making use of the large number of processing cores to efficiently parallelize calculations even for large state-size models. We implement the approach in an existing Bayesian framework and apply the algorithms to estimating the phylogeny of 62 complete mitochondrial genomes of carnivores under a 60-state codon model. We see a near 90-fold speed increase over an optimized CPU-based computation and a >140-fold increase over the currently available implementation, making this the first practical use of codon models for phylogenetic inference over whole mitochondrial or microorganism genomes. Source code provided in BEAGLE: Broad-platform Evolutionary Analysis General Likelihood Evaluator, a cross-platform/processor library for phylogenetic likelihood computation (http://beagle-lib.googlecode.com/). We employ a BEAGLE-implementation using the Bayesian phylogenetics framework BEAST (http://beast.bio.ed.ac.uk/).


Comparison of data from two vegetation monitoring methods in semi-natural grasslands.
Two vegetation-monitoring methods were compared: subplot frequency analysis (SF) and visual estimation of percentage cover (VE). Two independent observers collected data from two semi-natural, species-rich grasslands on three different occasions during the growth-season. During the last data collection period, survey times were also recorded. The two different data sets from the two methods were compared using partial Redundancy Analyses. The purpose of the comparison was to identify the method that explains most of the relevant variation in biodiversity-monitoring (inter- and intra-site variation), and the variation irrelevant when evaluating data (systematic inter-observer variation and variation due to phenological changes). Compared with VE data, more variation in SF data could be explained by spatial variables, while less variation depended on the observer and time of year surveyed. SF also found more species per plot but took on average five times longer to complete than VE. In conclusion, the different methods are suitable for different purposes: SF is more suitable for purposes demanding high accuracy and high precision, such as long-term biodiversity-monitoring when the identification of small changes has high priority, while VE might be more suitable for a one-time mapping of a large area.


Guideline-Directed Medication Use in Patients With Heart Failure With Reduced Ejection Fraction in India: American College of Cardiology's PINNACLE India Quality Improvement Program.
Little is known about the use of guideline-directed medical therapy (GDMT) in outpatients with heart failure with reduced left ventricular ejection fraction (HFrEF; ≤40%) in India. Our objective was to understand the use of GDMT in outpatients with HFrEF in India. The Practice Innovation And Clinical Excellence (PINNACLE) India Quality Improvement Program (PIQIP) is a registry for cardiovascular quality improvement in India supported by the American College of Cardiology Foundation. Between January 2008 and September 2014, we evaluated documentation of use of angiotensin-converting enzyme inhibitors (ACEIs)/angiotensin receptor blockers (ARBs) and β-blockers, or both, among outpatients with HFrEF seeking care in 10 centers enrolled in the PIQIP registry. Among 75 639 patients in the PIQIP registry, 34 995 had EF reported, and 15 870 had an EF ≤40%. The mean age was 56 years; 23% were female. Hypertension, diabetes, coronary artery disease, and myocardial infarction were present in 37%, 23%, 27%, and 17%, respectively. Use of ACEIs/ARBs, β-blockers, and both were documented in 33.5%, 34.9%, and 29.6% of patients, respectively. The documentation of GDMT was higher in men, in patients age ≥65 years, and in those with presence of hypertension, diabetes, or coronary artery disease. Documentation of GDMT gradually increased over the study period. Among patients enrolled in the PIQIP registry, about two-thirds of patients with EF ≤40% did not have documented receipt of GDMT. This study is an initial step toward improving adherence to GDMT in India and highlights the feasibility of examining quality of care in HFrEF in a resource-limited setting.


History of geriatric medicine: from Hippocrates to Marjory Warren.
It is widely assumed that geriatric medicine was an invention of the twentieth century. However, from the time of Hippocrates, there has been interest in the prolongation of the lifespan, the maintenance of health in old age and agerelated disease patterns. The debate about whether old age was a natural phenomenon or a disease state was not resolved until the nineteenth century. Calls for medicine relating to old age to be recognised as a discrete entity at the time when medical specialisation was developing were disregarded until the second half of the twentieth century. This review discusses the history of the theories of ageing and of disease and the practice of medicine for older people from the classical period up to Marjory Warren's initiative in London in 1935 and the development of geriatrics as a medical specialty.


Preclinical evaluation of the supercritical extract of azadirachta indica (neem) leaves in vitro and in vivo on inhibition of prostate cancer tumor growth.
Azadirachta indica, commonly known as neem, has gained worldwide prominence because of its medical properties, namely antitumor, antiviral, anti-inflammatory, antihyperglycemic, antifungal, and antibacterial activities. Despite these promising results, gaps remain in our understanding of the molecular mechanism of action of neem compounds and their potential for use in clinical trials. We investigated supercritical extract of neem leaves (SENL) for the following: molecular targets in vitro, in vivo efficacy to inhibit tumor growth, and bioactive compounds that exert antitumor activity. Treatment of LNCaP-luc2 prostate cancer cells with SENL suppressed dihydrotestosterone-induced androgen receptor and prostate-specific antigen levels. SENL inhibited integrin β1, calreticulin, and focal adhesion kinase activation in LNCaP-luc2 and PC3 prostate cancer cells. Oral administration of SENL significantly reduced LNCaP-luc2 xenograft tumor growth in mice with the formation of hyalinized fibrous tumor tissue, reduction in the prostate-specific antigen, and increase in AKR1C2 levels. To identify the active anticancer compounds, we fractionated SENL by high-pressure liquid chromatography and evaluated 16 peaks for cytotoxic activity. Four of the 16 peaks exhibited significant cytotoxic activity against prostate cancer cells. Mass spectrometry of the isolated peaks suggested the compounds with cytotoxic activity were nimbandiol, nimbolide, 2',3'-dihydronimbolide, and 28-deoxonimbolide. Analysis of tumor tissue and plasma samples from mice treated with SENL indicated 28-deoxonimbolide and nimbolide as the bioactive compounds. Overall, our data revealed the bioactive compounds in SENL and suggested that the anticancer activity could be mediated through alteration in androgen receptor and calreticulin levels in prostate cancer.


Kissing-Y stenting for endovascular treatment of complex wide necked bifurcation aneurysms using Acandis Acclino stents: results and literature review.
Y-configured stent assisted coiling is a promising therapeutic option to ensure safe coil embolization and preserve the affected arteries in complex wide necked aneurysms. We present our experience with self-expanding Acandis Acclino stents for the treatment of complex aneurysms using the kissing-Y technique. We retrospectively reviewed seven patients with seven complex aneurysms (three anterior communicating artery (AcomA), two middle cerebral artery, one basilar artery/superior cerebellar artery, and one vertebral artery/posterior inferior cerebellar artery) who were treated with the kissing-Y technique by stent assisted coiling from June 2013 to July 2014, with follow-up until January 2015. DSA follow-up was up to 17 months, with a mean follow-up period of 10 months. Six patients were treated electively and one in the acute phase of a subarachnoid hemorrhage. In all cases, closed cell Acandis Acclino stents were used. We evaluated procedural complications, clinical outcomes, and mid term angiographic follow-up. Additionally, a literature review is provided. In all patients, stents were successfully placed and implanted. One patient developed a periprocedural thromboembolic complication not directly related to the stents. No other periprocedural or postprocedural complications were encountered. Follow-up examinations showed stable and total occlusion of all coiled aneurysms. The results of our study show that the kissing-Y technique using closed cell Acandis Acclino stents followed by coil embolization is a feasible treatment option for selected complex bifurcation aneurysms.


Wearable Motion Sensor Device to Facilitate Rehabilitation in Patients With Shoulder Adhesive Capsulitis: Pilot Study to Assess Feasibility.
Adhesive capsulitis (AC) of the shoulder is a common disorder that painfully reduces the shoulder range of motion (ROM) among middle-aged individuals. Although physical therapy with home-based exercises is widely advised to restore ROM in the treatment of AC, clinical results vary owing to inconsistent patient compliance. In this study, we aimed to verify the feasibility of a treatment model that involves applying a wearable motion sensor device to assist patients conduct home-based exercises to improve training compliance and the accuracy of exercises, with the ultimate goal of improving the functional recovery of patients with AC. The motion sensor device was comprised of inertial measurement unit-based sensors and mobile apps for patients and physicians, offering shoulder mobility tracing, home-based exercise support, and progress monitoring. The interrater reliability of shoulder mobility measurement using the motion sensor device on 10 healthy participants and 15 patients with AC was obtained using an intraclass correlation coefficient analysis and compared with the assessments performed by two highly experienced physicians. A pilot prospective control trial was then carried out to allocate the 15 patients with AC to two groups: home-based exercise group and motion sensor-assisted rehabilitation group. Changes in active and passive shoulder ROM, pain and functional scores, and exercise completion rates were compared between the groups during a treatment period of 3 months. Shoulder ROM, as measured using the motion sensor device, exhibited good to excellent reliability based on the comparison with the measurements of two physicians (intraclass correlation coefficient range, 0.771 to 0.979). Compared with patients with AC in the home-based exercise group, those in the motion sensor-assisted rehabilitation group exhibited better shoulder mobility and functional recovery and a higher exercise completion rate during and after 3 months of rehabilitation. Motion sensor device-assisted home-based rehabilitation for the treatment of AC is a useful treatment model for telerehabilitation that enhances the compliance of patients through training, thus improving functional recovery. This helps overcome important obstacles in physiotherapy at home by providing comprehensible and easily accessible exercise instructions, enhancing compliance, ensuring the correctness of exercise, and monitoring the progress of patients.


The prevalence of Salmonella enteritidis and other Salmonella sp. among Canadian registered commercial chicken broiler flocks.
A nation-wide survey was conducted to estimate the prevalence of Salmonella enteritidis and other salmonellas among Canadian commercial broiler flocks. Environmental (litter and/or water) samples from 226 of 294 (76.9%) randomly selected flocks were contaminated with salmonellas. Litter samples were more often contaminated with salmonellas than water samples (47.4 v. 12.3%). Fifty different salmonella serovars were isolated. The most prevalent serovars were S. hadar, S. infantis, and S. schwarzengrund; they were isolated from samples of 98/294 (33.3%), 26/294 (8.8%), and 21/294 (7.1%) flocks, respectively. Feed samples of 39/290 (13.4%) flocks were contaminated with salmonellas. Salmonella enteritidis was isolated from the environmental samples of 9/294 (3.1%) flocks. Salmonella enteritidis phage type (PT) 8 was isolated from seven flocks, and PT 13a from two flocks.


Molecular evolution of imprinted genes: no evidence for antagonistic coevolution.
Genomically imprinted genes are those for which expression is dependent on the sex of the parent from which they are derived. Numerous theories have been proposed for the evolution of genomic imprinting: one theory is that it is an intra-individual manifestation of classical parent -offspring conflict. This theory is unique in predicting that an arms race may develop between maternally and paternally derived genes for the control of foetal growth demands. Such antagonistic coevolution may be mediated through changes in the structure of the proteins concerned. Comparable coevolution is the most likely explanation for the rapid changes seen in antigenic components of parasites and antigen recognition components of immune systems. We have examined the evolution of insulin-like growth factor Igf2, and its antagonistic receptor Igf2r) and find that in contrast to immune genes, at the sites of mutual binding they are highly conserved. In addition, we have analysed the rate of molecular evolution of seven imprinted genes including Igf2 and Igf2r), sequenced in both mouse and rat, and had that this is the same as that of nonimprinted receptors and significantly lower than that of immune genes controlling for differences in mutation rates. Contrary to the expectations of the conflict hypothesis, we hence find no evidence for antagonistic coevolution of imprinted genes mediated by changes in sequence.


Potentiation of 5-hydroxytryptamine (5-HT) responses by a 5-HT uptake inhibitor in pulmonary and systemic vessels: effects of exposing rats to hypoxia.
The aim was to determine whether uptake of 5-hydroxytryptamine (5-HT) by the 5-HT transporter (SERT) modulates contractile responses to 5-HT in rat pulmonary arteries and whether this modulation is altered by exposure of rats to chronic hypoxia (10% oxygen; 8 h/day; 5 days). The effects of the SERT inhibitor, citalopram (100 nM), on contractions to 5-HT were determined in isolated ring preparations of pulmonary artery (intralobar and main) and compared with data obtained in systemic arteries. In intralobar pulmonary arteries citalopram produced a potentiation (viz. an increase in potency, pEC(50)) of 5-HT. The potentiation was endothelium-dependent in preparations from normoxic rats but endothelium-independent in preparations from hypoxic rats. In main pulmonary artery endothelium-independent potentiation was seen in preparations from hypoxic rats but no potentiation occurred in preparations from normoxic rats. In systemic arteries, citalopram caused endothelium-independent potentiation in aorta but no potentiation in mesenteric arteries; there were no differences between hypoxic and normoxic rats. It is concluded that SERT can influence the concentration of 5-HT in the vicinity of the vasoconstrictor receptors in pulmonary arteries. The data suggest that in pulmonary arteries from hypoxic rats, unlike normoxic rats, the SERT responsible for this effect is not in the endothelium and, hence, is probably in the smooth muscle. The data are compatible with reports that, in the pulmonary circulation, hypoxia induces/up-regulates SERT, and hence increases 5-HT uptake, in vascular smooth muscle. The findings may have implications in relation to the suggested use of SERT inhibitors in the treatment of pulmonary hypertension.


Systems genomics of metabolic phenotypes in wild-type Drosophila melanogaster.
Systems biology is an approach to dissection of complex traits that explicitly recognizes the impact of genetic, physiological, and environmental interactions in the generation of phenotypic variation. We describe comprehensive transcriptional and metabolic profiling in Drosophila melanogaster across four diets, finding little overlap in modular architecture. Genotype and genotype-by-diet interactions are a major component of transcriptional variation (24 and 5.3% of the total variation, respectively) while there were no main effects of diet (<1%). Genotype was also a major contributor to metabolomic variation (16%), but in contrast to the transcriptome, diet had a large effect (9%) and the interaction effect was minor (2%) for the metabolome. Yet specific principal components of these molecular phenotypes measured in larvae are strongly correlated with particular metabolic syndrome-like phenotypes such as pupal weight, larval sugar content and triglyceride content, development time, and cardiac arrhythmia in adults. The second principal component of the metabolomic profile is especially informative across these traits with glycine identified as a key loading variable. To further relate this physiological variability to genotypic polymorphism, we performed evolve-and-resequence experiments, finding rapid and replicated changes in gene frequency across hundreds of loci that are specific to each diet. Adaptation to diet is thus highly polygenic. However, loci differentially transcribed across diet or previously identified by RNAi knockdown or expression QTL analysis were not the loci responding to dietary selection. Therefore, loci that respond to the selective pressures of diet cannot be readily predicted a priori from functional analyses.


Spectral color matching by means of minimax approximation.
A computer algorithm has been developed by means of a minimax approximation to approximate an absorption band by linear combination of several dyes. The minimax approximation always gives feasible solutions and smaller errors than a least-squares method. Suitable computer programs have been developed based on the linear programming theory, and same practical approximations have been demonstrated for spectrally matching colors of human skin, blue sky, and nonselective gray of 50% luminous reflectance with twelve component dyes.


Prolactin is not required for the development of severe chronic experimental autoimmune encephalomyelitis.
Predominance of multiple sclerosis (MS) in women, reductions of disease flares during pregnancy, and their increase in the postpartum period have suggested a hormonal influence on MS activity. The hormone prolactin (PRL) has long been debated as a potential immune-stimulating factor in several autoimmune disorders, including MS and its animal model experimental autoimmune encephalomyelitis (EAE). However, to date, no data clearly ascribe a pathogenic role to PRL in these diseases. Using PRL receptor-deficient (Prlr(-/-)) and PRL-deficient (Prl(-/-)) mice, we show that PRL plays a redundant role in the development of chronic EAE. In Prlr(-/-) and Prl(-/-) mice, EAE developed with a delayed onset compared with littermate control mice, but with full clinical severity. In line with the clinical outcome, T cell proliferation and production of IFN-γ, IL-17A, and IL-6 induced by myelin Ag were delayed in Prlr(-/-) and Prl(-/-) mice. Ag-specific IgG Ab responses were not affected by PRLR or PRL deficiency. We also show that mouse lymph node cells and purified CD4(+) T cells express transcript for Prlr, but not for Prl. These results reveal that PRL does not play a central role in the development of chronic EAE and optimal Th1 and Th17 responses against myelin. Moreover, they also rule out a possible contribution of PRL secreted by immune cells to the modulation of autoreactive T cell response in this model.


[Levels of low birth-weight in Europe: registration problems and effects of medical interventions].
The rate of low birthweight births, percentage of live births weighing less than 2500 grams is one of the perinatal health indicators recommended by the World Health Organisation. A review of the literature has revealed numerous problems related to this indicator. Some countries, including France and the Netherlands do not collect birthweight data. Elsewhere, there are more than 10% "unknown birthweight". The poorest registration is for birthweights below 1000 grams. Exclusion of babies who were registered as stillborn, when in fact they died shortly after birth can cause underregistration of low birthweights. Conversely, inclusion of true stillbirths will bring on an overestimation of low birthweight rate. Some countries have too few births to deliver accurate rates. Rates of medical interventions, such as infertility treatment and elective induction should be taken into account when analyzing differences between countries. Improvement in quality of registration and caution in the use of this indicator are warranted.


Listeria monocytogenes in Stone Fruits Linked to a Multistate Outbreak: Enumeration of Cells and Whole-Genome Sequencing.
In 2014, the identification of stone fruits contaminated with Listeria monocytogenes led to the subsequent identification of a multistate outbreak. Simultaneous detection and enumeration of L. monocytogenes were performed on 105 fruits, each weighing 127 to 145 g, collected from 7 contaminated lots. The results showed that 53.3% of the fruits yielded L. monocytogenes (lower limit of detection, 5 CFU/fruit), and the levels ranged from 5 to 2,850 CFU/fruit, with a geometric mean of 11.3 CFU/fruit (0.1 CFU/g of fruit). Two serotypes, IVb-v1 and 1/2b, were identified by a combination of PCR- and antiserum-based serotyping among isolates from fruits and their packing environment; certain fruits contained a mixture of both serotypes. Single nucleotide polymorphism (SNP)-based whole-genome sequencing (WGS) analysis clustered isolates from two case-patients with the serotype IVb-v1 isolates and distinguished outbreak-associated isolates from pulsed-field gel electrophoresis (PFGE)-matched, but epidemiologically unrelated, clinical isolates. The outbreak-associated isolates differed by up to 42 SNPs. All but one serotype 1/2b isolate formed another WGS cluster and differed by up to 17 SNPs. Fully closed genomes of isolates from the stone fruits were used as references to maximize the resolution and to increase our confidence in prophage analysis. Putative prophages were conserved among isolates of each WGS cluster. All serotype IVb-v1 isolates belonged to singleton sequence type 382 (ST382); all but one serotype 1/2b isolate belonged to clonal complex 5. WGS proved to be an excellent tool to assist in the epidemiologic investigation of listeriosis outbreaks. The comparison at the genome level contributed to our understanding of the genetic diversity and variations among isolates involved in an outbreak or isolates associated with food and environmental samples from one facility. Fully closed genomes increased our confidence in the identification and comparison of accessory genomes. The diversity among the outbreak-associated isolates and the inclusion of PFGE-matched, but epidemiologically unrelated, isolates demonstrate the high resolution of WGS. The prevalence and enumeration data could contribute to our further understanding of the risk associated with Listeria monocytogenes contamination, especially among high-risk populations.