Interleukin-1 beta and phenytoin reduce alpha 1 (I) procollagen mRNA expression in human gingival fibroblasts.
Effects of and interactions between interleukin-1 beta (IL-1 beta) and phenytoin (PHT) on alpha 1 (I) procollagen gene and protein expression in human gingival fibroblasts and its relation to prostaglandin E2 (PGE2) formation were studied. IL-1 beta (300 pg/ ml) reduced the steady-state level of alpha 1 (I) procollagen mRNA by 50% and decreased the amount of procollagen I by 35%. PHT (10 micrograms/ml) reduced the level of alpha 1 (I) procollagen mRNA by 40% but the amount of procollagen I in the medium was unchanged. In combination with IL-1 beta, PHT potentiated the inhibitory effect of IL-1 beta on alpha 1 (I) procollagen mRNA level that was accompanied by an increased PGE2 formation. Preincubation with indomethacin (10(-6) M) partially reduced the inhibitory effect of IL-1 beta as well as of IL-1 beta in combination with PHT on the mRNA level of alpha 1 (I) procollagen. The inhibitory effect of PHT was unaffected by indomethacin treatment. Addition of exogenous PGE2 (> or = 10 nM) dose-dependently reduced steady-state level of alpha 1 (I) procollagen mRNA as well as the amount of procollagen 1. The study indicates that IL-1 reduces the expression of alpha 1 (I) procollagen mRNA in human gingival fibroblasts partly by a prostaglandin endoperoxide (PGH) synthase-mediated pathway and partly by a PGH-synthase independent pathway, whereas PHT reduces alpha 1 (I) procollagen gene expression by a PGH-synthase independent pathway. The potentiation of the inhibitory effect of IL-1 induced by PHT was mediated mainly by a PGH-synthase dependent pathway.


Developmental switches in chemokine response profiles during B cell differentiation and maturation.
Developing B cells undergo dramatic changes in their responses to chemoattractant cytokines (chemokines) and in expression of chemokine receptors. Bone marrow pre-pro-B cells (AA4.1(+)/natural killer 1.1(-) Fraction A cells) and cells capable of generating pro-B colonies in the presence of interleukin 7 and flt3 ligand migrate to thymus-expressed chemokine (TECK), a response lost in later stages of B cell development. B cell-attracting chemokine 1 (BCA-1) responses correlate with CXC chemokine receptor (CXCR)5 expression, are first displayed by a pro-B cell subset, are lost in pre-B cells, and then are regained just before and after egress from the marrow. All peripheral B cell subsets, including follicular and germinal center as well as marginal zone and peritoneal B1 B cells, respond to BCA-1, implying that responsiveness to this follicular chemokine is not sufficient to predict follicle localization. Responses to the CC chemokine receptor (CCR)7 ligands secondary lymphoid tissue chemoattractant (SLC) and macrophage inflammatory protein (MIP)-3beta, implicated in homing to lymphoid tissues, are upregulated before B cell exit from the marrow, but increase further in the periphery and are shared by all peripheral B cells. In contrast, responsiveness to MIP-3alpha and expression of CCR6 are acquired only after emigration to the periphery and during maturation into the recirculating B cell pool. Chemotaxis to stromal cell-derived factor 1alpha is observed at all stages of B cell differentiation. Thus, unique patterns of chemokine responses may help define developing B cell populations and direct their maturation in the marrow and migration to the periphery.


[The impact of maternal education on the dietary and nutritional status of preschool children--a case study in 8 provinces of China].
This analysis is based on the data from China Health and Nutrition Survey in collaboration with University of North Carolina at Chapel Hill undertaken in 8 provinces of China from 1993 to 1996. Totally, 1098 mother-child pairs under age 6 were investigated. Descriptive analysis and stratified analysis are applied to investigate the impact of maternal education on the dietary and nutritional status of children. Both the crude analysis and after controlling for confounding variables, maternal education in the population remains statistically relevant to children's dietary and nutritional status. The proportion of low body weight and stunting children is the highest in the group with maternal education level being below preliminary school, the lowest in the group with education level being above high middle school. Moreover, in these three groups, the higher the level of maternal education, the greater the percentage of nutrients intake of China RDA, especially those from animal food. These results fully illustrate the importance of maternal education for the prevention of malnutrition in children.


Inactivation of RASSF1C during in vivo tumor growth identifies it as a tumor suppressor gene.
RASSF1A, a major member of the RASSF1 gene family, is silenced by promoter methylation at a high frequency in a large number of human solid tumors. Controlled expression of RASSF1A reverts the tumorigenic phenotype of several human cancer cell lines. Here we investigated another main isoform, RASSF1C, and compared it with RASSF1A in the gene inactivation test (GIT), based on a tetracycline regulation system. In the small-cell lung cancer (SCLC) line U2020, only RASSF1A has shown growth inhibitory activity in vitro, while in the prostate cell line LNCaP and renal cell carcinoma (RCC) line KRC/Y both RASSF1A and RASSF1C showed similar (approximately 90%) suppressing activity in vitro. Both RASSF1C and RASSF1A suppressed the tumorigenicity of the KRC/Y RCC cell line in SCID mice. Mutations, deletions and loss of expression of RASSF1A and RASSF1C transgenes were identified in all 15 grown SCID tumors. In contrast, the mutant RASSF1A containing Cys65Arg and Val211Ala had reduced growth suppression activity both in vitro and in vivo and did not show any further changes in four grown SCID tumors. In addition, RASSF1C was shown to induce cell cycle arrest in KRC/Y cells. These results strongly imply that like RASSF1A the RASSF1C gene could serve a tumor suppressor function.


Curcumin dietary supplementation ameliorates disease phenotype in an animal model of Huntington's disease.
Huntington's disease (HD) has traditionally been described as a disorder purely of the brain; however, evidence indicates that peripheral abnormalities are also commonly seen. Among others, severe unintended body weight loss represents a prevalent and often debilitating feature of HD pathology, with no therapies available. It correlates with disease progression and significantly affects the quality of life of HD patients. Curcumin, a naturally occurring polyphenol with multiple therapeutic properties, has been validated to exert important beneficial effects under health conditions as well as in different pathological settings, including neurodegenerative and gastrointestinal (GI) disorders. Here, we investigated the potential therapeutic action that curcumin-supplemented diet may exert on central and peripheral dysfunctions in R6/2 mice, a well-characterized HD animal model which recapitulates some features of human pathology. Maintenance of normal motor function, protection from neuropathology and from GI dysfunction and preservation of GI emptying and conserved intestinal contractility, proved the beneficial role of life-long dietary curcumin in HD and corroborated the potential of the compound to be exploited to alleviate very debilitating symptoms associated with the disease.


Glucose loading prevents freezing injury in rapidly cooled wood frogs.
The wood frog (Rana sylvatica) is the most commonly studied of ten species of freeze-tolerant vertebrates. Under natural (i.e., slow) rates of cooling, freezing initiates the production of the cryoprotectant glucose, which is mobilized from the liver and distributed to tissues throughout the body. Rapid cooling during freezing is injurious to wood frogs, probably because cryoprotectant production and mobilization are inhibited. To test this hypothesis, we investigated whether rapid-cooling injury is reduced if exogenous glucose is experimentally introduced to tissues before freezing. Glucose-loaded and control (saline-injected) wood frogs were rapidly cooled during freezing to -2.5 degrees C and subsequently assayed for injury at both cellular (erythrocyte) and neuromuscular (behavioral reflex) levels. Rapid cooling produced substantial hemolysis in control frogs, but erythrocyte injury was significantly reduced in glucose-loaded frogs. Similarly neuromuscular injury was significantly higher in control frogs than in glucose-loaded frogs. These findings suggest that rapid-cooling injury results from an inadequate production and distribution of endogenous glucose during freezing. Furthermore, the inverse relationship between the degree of freezing injury and the quantity of exogenous glucose present strongly implicates glucose as a cryoprotectant in R. sylvatica.


A genome search identifies major quantitative trait loci on human chromosomes 3 and 4 that influence cholesterol concentrations in small LDL particles.
Small, dense LDL particles are associated with increased risk of cardiovascular disease. To identify the genes that influence LDL size variation, we performed a genome-wide screen for cholesterol concentrations in 4 LDL size fractions. Samples from 470 members of randomly ascertained families were typed for 331 microsatellite markers spaced at approximately 15 cM intervals. Plasma LDLs were resolved by using nondenaturing gradient gel electrophoresis into 4 fraction sizes (LDL-1, 26.4 to 29.0 nm; LDL-2, 25.5 to 26.4 nm; LDL-3, 24.2 to 25.5 nm; and LDL-4, 21.0 to 24.2 nm) and cholesterol concentrations were estimated by staining with Sudan Black B. Linkage analyses used variance component methods that exploited all of the genotypic and phenotypic information in the large extended pedigrees. In multipoint linkage analyses with quantitative trait loci for the 4 fraction sizes, only LDL-3, a fraction containing small LDL particles, gave peak multipoint log10 odds in favor of linkage (LOD) scores that exceeded 3.0, a nominal criterion for evidence of significant linkage. The highest LOD scores for LDL-3 were found on chromosomes 3 (LOD=4.1), 4 (LOD=4.1), and 6 (LOD=2.9). In oligogenic analyses, the 2-locus LOD score (for chromosomes 3 and 4) increased significantly (P=0.0012) to 6.1, but including the third locus on chromosome 6 did not significantly improve the LOD score (P=0.064). Thus, we have localized 2 major quantitative trait loci that influence variation in cholesterol concentrations of small LDL particles. The 2 quantitative trait loci on chromosomes 3 and 4 are located in regions that contain the genes for apoD and the large subunit of the microsomal triglyceride transfer protein, respectively.


[The relation between metabolism of biopterin and dystonia-parkinsonism].
Tetrahydrobiopterin (BH4) is an essential cofactor for tyrosine hydroxylase. BH4 can be synthesized from GTP through three enzymatic reactions. The rate-limiting step of the BH4 synthesis is catalyzed by GTP cyclohydrolase I (GCH). Recently, we found that GCH is a causative gene for hereditary progressive dystonia/dopa-responsive dystonia (HPD/DRD). However, several problems still remain to be solved. The first concern is the presence of asymptomatic carriers in the disease. The difference between symptomatic and asymptomatic carriers is unknown. Second, we cannot find any mutation in the coding region of the GCH gene in about 40% of the patients. What kind of mutation would be present in these patients. The last concern is the molecular mechanism how the enzymatic activity is decreased to less than 20% of normal values. Further studies are required to solve the questions.


Recreational physical activity and leisure-time sitting in relation to postmenopausal breast cancer risk.
Epidemiologic evidence supports an inverse association between physical activity and postmenopausal breast cancer. Whether associations exist for moderate activities, such as walking, and whether associations differ by estrogen receptor (ER) status, body mass index (BMI, kg/m(2)), adult weight gain, or use of postmenopausal hormones (PMH) is unclear. The relation between time spent sitting and breast cancer also is unclear. Among 73,615 postmenopausal women in the American Cancer Society Cancer Prevention Study II Nutrition Cohort, 4,760 women were diagnosed with breast cancer between 1992 and 2009. Extended Cox regression was used to estimate multivariable-adjusted relative risks (RR) of breast cancer in relation to total recreational physical activity, walking, and leisure-time sitting. Differences in associations by ER status, BMI, weight gain, and PMH use were also evaluated. The most active women (those reporting >42 MET-hours/week physical activity) experienced 25% lower risk of breast cancer than the least active [0-<7 MET-hours/week; 95% confidence interval (CI), 0.63-0.89; Ptrend = 0.01]. Forty-seven percent of women reported walking as their only recreational activity; among these women, a 14% lower risk was observed for ≥7 hours/week relative to ≤3 hours/week of walking (95% CI, 0.75-0.98). Associations did not differ by ER status, BMI, weight gain, or PMH use. Sitting time was not associated with risk. These results support an inverse association between physical activity and postmenopausal breast cancer that does not differ by ER status, BMI, weight gain, or PMH use. The finding of a lower risk associated with ≥7 hours/week of walking may be of public health interest.


Contribution of horizontal gene transfer to the emergence of VIM-4 carbapenemase producer Enterobacteriaceae in Kuwait.
Carbapenem-resistant Enterobacteriaceae encountered in countries of the Arabian Peninsula usually produce OXA-48-like and New Delhi metallo-beta-lactamases (NDM) carbapenemases. However, a temporary increase in VIM-4-producing, clonally unrelated Enterobacteriaceae strains was described earlier in a Kuwaiti hospital. We investigated the genetic support of blaVIM-4 in six Klebsiella pneumoniae strains, one Escherichia coli, and one Enterobacter cloacae strain and compared it to that of VIM-4-producing isolates from other countries of the region. Five K. pneumoniae strains and the E. coli strain from Kuwait carried an ~165 kb IncA/C-type plasmid indistinguishable by restriction fragment length polymorphism. The complete sequence of one of them (pKKp4-VIM) was established. pKKp4-VIM exhibited extensive similarities to episomes pKP-Gr642 carrying blaVIM-19 encountered in Greece and to the partially sequenced pCC416 harboring blaVIM-4 detected in Italy. In other countries of the region, the only similar plasmid was the one detected in the isolate from the UAE. In all Kuwaiti strains, irrespective of the species and their VIM plasmids, the blaVIM-4 gene was located within the same integron structure (In416), different from those of other countries of the region. Our data show that the spread of this IncA/C plasmid and particularly that of the In416 integron caused a considerable, albeit temporary, increase in the rate of mostly clonally unrelated VIM-producing Enterobacteriaceae strains of multiple species. Monitoring of such events is of high importance as the interference with the spread of mobile genetic elements may represent a formidable challenge to infection control.


Immunoaffinity techniques coupled to mass spectrometry for the analysis of human peptide hormones: advances and applications.
The accurate and comprehensive determination of peptide hormones from biological fluids has represented a considerable challenge to analytical chemists for decades. Besides long-established bioanalytical ligand binding assays (or ELISA, RIA, etc.), more and more mass spectrometry-based methods have been developed recently for purposes commonly referred to as targeted proteomics. Eventually the combination of both, analyte extraction by immunoaffinity and subsequent detection by mass spectrometry, has shown to synergistically enhance the test methods' performance characteristics. Areas covered: The review provides an overview about the actual state of existing methods and applications concerning the analysis of endogenous peptide hormones. Here, special focus is on recent developments considering the extraction procedures with immobilized antibodies, the subsequent separation of target analytes, and their detection by mass spectrometry. Expert commentary: Key aspects of procedures aiming at the detection and/or quantification of peptidic analytes in biological matrices have experienced considerable improvements in the last decade, particularly in terms of the assays' sensitivity, the option of multiplexing target compounds, automatization, and high throughput operation. Despite these advances and progress as expected to be seen in the near future, immunoaffinity purification coupled to mass spectrometry is not yet a standard procedure in routine analysis compared to ELISA/RIA.


Targeted sequencing identifies risk variants in 202 candidate genes for neurodevelopmental disorders.
With the continuous deepening of genetic research on neurodevelopmental disorders (NDDs), more patients have been identified the causal or candidate genes. However, it is still urgent needed to increase the sample size to confirm the associations between variants and clinical manifestations. We previously performed molecular inversion probe sequencing of autism spectrum disorder (ASD) candidate genes in 1,543 ASD patients. In this study, we used the same method to detect de novo variants (DNVs) in 665 NDD patients with intellectual disability (ID) and/or epilepsy (EP) for genetic analysis and diagnosis. We compared findings from ID/EP and ASD patients to improve our understanding of different subgroups of NDDs. We identified 72 novel variants and 39 DNVs. A totally of 5.71% (38/665) of the patients were genetically diagnosed by this sequencing strategy. ID/EP patients demonstrated a higher prevalence of likely gene disruptive DNVs in ASD genes than the healthy population. Regarding high-risk genes, SCN1A and CKDL5 were more frequently mutated in ID/EP patients than in ASD patients. Our data provide an overview of the mutation burden in ID/EP patients from the perspective of high risk ASD genes, indicating the differences and association of NDDs subgroups.


Distribution of gamma-carboxyglutamic acid residues in partially carboxylated human prothrombins.
The role of gamma-carboxyglutamic acid in prothrombin has been examined using partially carboxylated variant prothrombins isolated from a person with a hereditary defect in vitamin K-dependent carboxylation. These species differ in gamma-carboxyglutamic acid content, distribution, and function, as monitored by metal binding properties, conformational transitions, phospholipid binding, and calcium-dependent coagulant activity (Borowski, M., Furie, B. C., Goldsmith, G. H., and Furie, B. (1985) J. Biol. Chem. 260, 9258-9264). The distribution of gamma-carboxyglutamic acids in the variant prothrombin species was determined by specific tritium incorporation into gamma-carboxyglutamic acid residues, thermal decarboxylation, and automated Edman degradation. gamma-Carboxyglutamic acid residues in the partially carboxylated prothrombins were identified by the assay of tritium in the resultant glutamic acid residues in the acarboxyprothrombins. The results indicate that variant prothrombins 1-3 are nearly homogeneous populations of partially carboxylated prothrombins. The ability of prothrombin to undergo a metal-induced conformational change and to bind to phospholipid vesicles correlated closely to the presence of a gamma-carboxyglutamic acid at residue 16. This residue is likely involved in the formation of a critical high affinity metal-binding site, possibly formed by Gla 16 and Gla 25 and/or Gla 26. A second high affinity metal-binding site, present in all of the variant prothrombin species, is defined, as an upper limit, by Gla 6, Gla 14, Gla 19, and Gla 20. This region is likely responsible for the interaction of certain of the conformation-specific antibodies to the metal-stabilized conformer of prothrombin.


Synthesis of indolo[2,1-a]isoquinoline derivatives via visible-light-induced radical cascade cyclization reactions.
We describe a photocatalyzed transformation for the synthesis of the indolo[2,1-a]isoquinoline core structure. This redox neutral reaction features mild reaction conditions and exceptional functional group tolerance. A series of valuable indolo[2,1-a]isoquinoline derivatives bearing various functional groups were synthesized using this method in good to excellent yields.


Clinical roundtable monograph: safety and efficacy of lipid-based amphotericin B.
Invasive fungal infections are an important cause of morbidity and mortality in critically ill and immunocompromised patients. Amphotericin B has remained the gold standard treatment for systemic fungal infections. The primary obstacle to amphotericin B therapy is its poor solubility and dose-related toxicities, especially renal impairment. As a result, newer lipid-based formulations of amphotericin B have been developed. This monograph compares available phamacokinetic, efficacy, and safety data for the 3 lipid-based formulations of amphotericin B, with reference to various patient populations. Potential causes of infusion-related reactions are described, and a premedication algorithm for the prevention of infusion-related reactions is developed.


[Targeting of anthracyclines and hepatomas].
DNR can be targeted to the liver by linking to galactosylated human serum albumin (AG). The linkage is stable in the blood stream and allows the release of active DNR after endocytosis of the conjugate by the target cells. Receptors for AG are present at the cell membrane of hepatocytes, primary human hepatoma cells and lung metastases. After i.v. administration of AG-DNR more than 70% of the drug is taken up by the liver and in rats 50% of DNR is eliminated in the bile after 16 h. Nude mice bearing human hepatoblastoma cells implanted s.c. were treated twice a week with a dose equivalent to 6.6 mg/kg of DNR either for the free drug or the conjugate AG-DNR. After 7 injections, tumor growth is inhibited in both case, however, the conjugate seems more active and is at least twice less toxic in terms of LD50. A phase I clinical trial of AG-DNR on 11 patients bearing hepatocarcinoma revealed that despite a transitory hyperthermia (37-38 degrees C) during the first day of the 4 day-perfusion, and modifications of hepatic enzymes in 3 cirrhotic patients, no hematologic and cardiac toxicity could be detected. A subjective response has been obtained in half of the patients with a decrease of plasmatic alpha-foetoprotein levels by more than 50% in 4 patients and one complete remission of more than 23 months with disparition of pulmonary metastases.


Spermiogenesis in phytophagous bug (Hemiptera, Pentatomidae): an ultrastructural study.
Ultrastructural and immunocytochemical studies were carried out on spermiogenesis in two species of phytophagous bug, Acrosternum aseadum and Nezara viridula. The nucleus development involved changes in the shape and in the degree of chromatin condensation, with specific aggregation patterns of DNA-histone complex occurring during this process. The acrosome showed a three-layered and the acrosomal content a paracrystalline arrangement. The axoneme presented a 9 + 9 + 2 microtubule pattern and bridges occurred between axonemal microtubules 1 and 5, and mitochondrial derivatives. Two paracrystalline structures embedded in amorphous regions were observed in the mitochondrial derivatives. The use of the negative staining technique shows a zig-zag profile in the mitochondria due to infolding to the cristae, regularly spaced with approximately 40 nm. An electron dense rod was observed near the centriolar adjunct; it presented labelling for tubulin suggesting that this structure may be involved in the microtubule organization during spermiogenesis of these insects.


Muscle responses to transcranial stimulation in man depend on background oscillatory activity.
Muscle responses to transcranial stimulation show high sweep-to-sweep variability, which may reflect an underlying noise process in the motor system. We examined whether response amplitude correlated with the level of prestimulus background EMG, and network oscillations. Transcranial magnetic or electrical stimulation was delivered to primary motor cortex whilst human subjects performed a precision grip task known to promote beta-band ( approximately 20 Hz) cortical oscillations. Responses were recorded from two intrinsic hand muscles. Response magnitude correlated significantly with the level of background EMG (mean r(2) = 0.20). Using a novel wavelet method, we quantified the amplitude and phase of oscillations in prestimulus sensorimotor EEG. Surprisingly, response magnitude showed no significant correlation with EEG oscillations at any frequency. However, oscillations in the prestimulus EMG were significantly correlated with response size; the correlation coefficient had peaks around 20 Hz. When oscillations in one muscle were used to predict response amplitude in a different muscle, correlations were substantially smaller. Finally, for each recording, we calculated the best possible prediction of response size obtainable from up to 20 measures of prestimulus EEG and EMG oscillations. Such optimal predictions had low correlation coefficients (mean r(2) = 0.2; 76% were below 0.3). We conclude that prestimulus oscillations, mainly in the beta-band, do explain some of the variability in responses to transcranial stimulation. Oscillations may likewise increase the noise of natural motor processing, explaining why this form of network activity is usually suppressed prior to dynamic movements. However, the majority of the variation is determined by other factors, which are not accessible by noninvasive recordings.


Genetic differences in naloxone enhancement of ethanol-induced conditioned taste aversion.
The influence of the opioid system on acquisition of an ethanol-induced conditioned taste aversion was examined in alcohol-preferring and avoiding inbred strains of mice (C57BL/6J and DBA/2J). Fluid-deprived mice from each strain received either ethanol alone, naloxone alone, or both ethanol and naloxone immediately after access to a novel tasting fluid. Naloxone alone (1 or 3 mg/kg) did not induce a conditioned taste aversion in either strain of mice. Administration of ethanol (1.5 g/kg) to DBA/2J mice produced a moderate taste aversion that was not affected by co-administration of naloxone. Although ethanol administered alone (3 g/kg) did not cause a taste aversion in C57BL/6J mice, the combination of ethanol and the higher dose of naloxone produced a significant taste aversion that increased across trials. A second experiment addressed the possibility that naloxone failed to enhance the ethanol-induced condition taste aversion in DBA/2J mice due to a "floor" effect on consumption. A lower ethanol dose (1 g/kg) was given alone or in combination with naloxone (1 or 3 mg/kg). Again, ethanol produced a moderate conditioned taste aversion that was not potentiated by naloxone. Subsequent conditioning with a high ethanol dose produced further suppression of intake, confirming that naloxone's failure to enhance aversion on earlier trials was not due to a "floor" effect. These data demonstrate a strain specific interaction between the aversive effect of ethanol and naloxone. More specifically, the results indicate that blockade of opioid receptors enhances the aversive effect of ethanol in C57BL/6J but not DBA/2J mice, suggesting that genetically determined differences in the endogenous opioid system of alcohol-preferring mice may mitigate ethanol's aversive effect.


Visual excitation and blood clotting.
Both processes involve extremely large amplification between the stimulus and the response. In vision it had been suggested that thismight be achieved by a chain of successive proenzyme-enzyme activations. Such a chain has now been found to underlie the mechanism of blood clotting. Methods are suggested for pursuing this comparison further.


Nucleolus, ribosomes, and cancer.
The complex aspects linking the nucleolus and ribosome biogenesis to cancer are reviewed here. The available evidence indicates that the morphological and functional changes in the nucleolus, widely observed in cancer tissues, are a consequence of both the increased demand for ribosome biogenesis, which characterizes proliferating cells, and the changes in the mechanisms controlling cell proliferation. In fact, the loss or functional changes in the two major tumor suppressor proteins pRB and p53 cause an up-regulation of ribosome biogenesis in cancer tissues. In this context, the association in human carcinomas of nucleolar hypertrophy with bad prognoses is worthy of note. Further, an increasing amount of data coming from studies on both hepatitis virus-induced chronic liver diseases and a subset of rare inherited disorders, including X-linked dyskeratosis congenita, suggests an active role of the nucleolus in tumorigenesis. Both an up-regulation of ribosome production and changes in the ribosome structure might causally contribute to neoplastic transformation, by affecting the balance of protein translation, thus altering the synthesis of proteins that play an important role in the genesis of cancer.


Inhibition of AP-1 DNA binding by nitric oxide involving conserved cysteine residues in Jun and Fos.
Nitric oxide (NO), which has diverse biological effects, can modulate AP-1 activity. Since DNA binding of Jun-Jun and Jun-Fos dimers is regulated in vitro by redox control involving conserved cysteines, we hypothesized that the action of NO is mediated via these residues. We performed electrophoretic mobility-shift analyses using Jun and Fos recombinant proteins and NO solutions. Cysteine-to-serine mutants showed that the inhibition of AP-1 activity following NO treatment was dependent on the presence of Cys7272 and Cys154 in the DNA binding domain of Jun and Fos, respectively. The inhibitory effect of NO was reversed by DTT and the thioredoxin system. Our results demonstrate that NO mediates its inhibitory effect by reacting specifically with the conserved cysteine residues in Jun and Fos.


Of mice and men: tyrosinase modification of congenital glaucoma in mice but not in humans.
Primary congenital glaucoma (PCG) is an autosomal recessive ocular trait caused by mutations in the gene for cytochrome P4501B1 (CYP1B1). Although PCG is often considered to be fully penetrant, the disease shows 50% penetrance in some Saudi Arabian families. The familial segregation of the nonpenetrance suggests a genetic modifier. Recently, tyrosinase (Tyr) deficiency was found to worsen the drainage structure/ocular dysgenesis phenotype of Cyp1b1-/- mice, suggesting that Tyr is a modifier of the phenotype. In the current study, tyrosinase (TYR) was investigated in human PCG. A genome-wide screen, a single nucleotide polymorphism (SNP) analysis in the TYR chromosomal region 11q13-q21, and sequencing of the TYR gene was performed with individuals from Saudi Arabian families with multiple, clinically confirmed, molecularly proven, nonpenetrant members. The study outcome did not support TYR as a modifier of the PCG phenotype in this population. The sequencing data showed no TYR mutations in the nonpenetrant family members and no difference in polymorphism frequencies between nonpenetrant or fully penetrant families. TYR is not a modifier of the CYP1B1-associated PCG phenotype in the Saudi Arabian population.


Senescence and learning in honeybee (Apis mellifera) workers.
Foraging by honeybee workers was investigated from the moment of the first foraging flight until death. To minimise the influence of factors other than senescence the foragers were trained to collect food from an artificial flower close to their hive. During each foraging trip the workers repeatedly visited an artificial flower, collecting one microlitre of 50% sugar solution per visit. During the first 50 flights the mean time taken to collect one portion of food decreased significantly and the number of visits to the artificial flower per flight increased significantly. During flights following the 50th flight, the mean time taken to collect one portion of food increased significantly and the number of visits to the artificial flower per flight decreased significantly. The results confirm earlier observations that the foraging behaviour of honeybee workers is not only influenced by learning, but also by the effects of senescence.


alpha1,4-Glucosidase-albumin polymers: in vitro properties and advantages for enzyme replacement therapy.
Soluble polymers of rat (or human) albumin and alpha-1,4-glucosidase are prepared using the cross-linking agent glutaraldehyde. The resulting polymer has an average molecular weight of 800 000 indicating an average composition of 12 albumin molecules for each enzyme molecule. Compared with an equivalent amount of free enzyme, the enzyme-albumin polymer has an increased resistance to heat denaturation (half-life of 15 h compared with 1 h for free enzyme at 37 degrees C) and to proteolysis by trypsin (half-life of 180 min compared with 10 min). The high degree of resistance to bioinactivation of the enzyme-albumin polymer is discussed in relation to requirements for enzyme replacement therapy in a range of metabolic diseases including type II glycogenosis (Pompe's disease) where alpha-1,4-glucosidase is the defective enzyme.


Anorectal malformations and their associated anomalies.
Congenital malformation of the rectum and anus is a problem frequently encountered in the neonatal period. 88 patients presenting with anorectal malformations were analysed and physical, operative, radiological and autopsy findings were reviewed to determine the types and incidence of associated lesions. Genitourinary malformations form the major group of associated lesions and contribute a significant degree of morbidity in the overall management of the anorectal deformities.


Engaging community pharmacists in the primary prevention of cardiovascular disease: protocol for the Pharmacist Assessment of Adherence, Risk and Treatment in Cardiovascular Disease (PAART CVD) pilot study.
Cardiovascular disease (CVD) is the leading cause of death globally. Community pharmacist intervention studies have demonstrated clinical effectiveness for improving several leading individual CVD risk factors. Primary prevention strategies increasingly emphasise the need for consideration of overall cardiovascular risk and concurrent management of multiple risk factors. It is therefore important to demonstrate the feasibility of multiple risk factor management by community pharmacists to ensure continued currency of their role. This study will be a longitudinal pre- and post-test pilot study with a single cohort of up to 100 patients in ten pharmacies. Patients aged 50-74 years with no history of heart disease or diabetes, and taking antihypertensive or lipid-lowering medicines, will be approached for participation. Assessment of cardiovascular risk, medicines use and health behaviours will be undertaken by a research assistant at baseline and following the intervention (6 months). Validated interview scales will be used where available. Baseline data will be used by accredited medicines management pharmacists to generate a report for the treating community pharmacist. This report will highlight individual patients' overall CVD risk and individual risk factors, as well as identifying modifiable health behaviours for risk improvement and suggesting treatment and behavioural goals. The treating community pharmacist will use this information to finalise and implement a treatment plan in conjunction with the patient and their doctor. Community pharmacists will facilitate patient improvements in lifestyle, medicines adherence, and medicines management over the course of five counselling sessions with monthly intervals. The primary outcome will be the change to average overall cardiovascular risk, assessed using the Framingham risk equation. This study will assess the feasibility of implementing holistic primary CVD prevention programs into community pharmacy, one of the most accessible health services in most developed countries. Australia and New Zealand Clinical Trial Registry Number: ACTRN12609000677202.


Genetic analysis of the postsynaptic transmembrane X-linked neuroligin 3 gene in autism.
Autism is a complex neurodevelopmental disorder, the prevalence of which has increased drastically in India in recent years. Neuroligin is a type I transmembrane protein that plays a crucial role in synaptogenesis. Alterations in synaptic genes are most commonly implicated in autism and other cognitive disorders. The present study investigated the neuroligin 3 gene in the Indian autistic population by sequencing and in silico pathogenicity prediction of molecular changes. In total, 108 clinically described individuals with autism were included from the North Karnataka region of India, along with 150 age-, sex-, and ethnicity-matched healthy controls. Genomic DNA was extracted from peripheral blood, and exonic regions were sequenced. The functional and structural effects of variants of the neuroligin 3 protein were predicted. One coding sequence variant (a missense variant) and four non-coding variants (two 5'-untranslated region [UTR] variants and two 3'-UTR variants) were recorded. The novel missense variant was found in 25% of the autistic population. The C/C genotype of c.551T>C was significantly more common in autistic children than in controls (p = 0.001), and a significantly increased risk of autism (24.7-fold) was associated with this genotype (p = 0.001). The missense variant showed pathogenic effects and high evolutionary conservation over the functions of the neuroligin 3 protein. In the present study, we reported a novel missense variant, V184A, which causes abnormal neuroligin 3 and was found with high frequency in the Indian autistic population. Therefore, neuroligin is a candidate gene for future molecular investigations and functional analysis in the Indian autistic population.


Role of angiotensin II in potassium-mediated stimulation of aldosterone secretion in the dog.
Potassium is known to enhance the aldosterone-stimulating action of angiotensin II. Such a synergistic interaction of potassium with angiotensin II could represent an action by angiotensin II to potentiate potassium as a stimulus. To examine for this effect of angiotensin II on potassium, plasma aldosterone levels were measured before and after an infusion of potassium chloride (15 meq i.v.) into dogs without and with prevention of angiotensin II formation by captopril, an angiotensin converting-enzyme inhibitor. In addition, responses to potassium were measured in a group of dogs receiving angiotensin II plus captopril. After potassium infusion, control dogs showed an increase of 7.7 +/- 1.9 (SEM) ng/dl (P less than 0.001) in the level of plasma aldosterone. In contrast, captopril-treated dogs showed no change in plasma aldosterone concentration in response to potassium. When angiotensin II was administered to captopril-treated dogs responsiveness to potassium administration was restored (plasma aldosterone concentration increased by 7.4 +/- 2.1 ng/dl, P less than 0.002). ACTH stimulated aldosterone secretion despite captopril treatment (P less than 0.001), however, ACTH produced a greater increase in the plasma aldosterone concentration in controls than in captopril-treated animals. It is evident from these results that stimulation of aldosterone secretion by potassium is considerably enhanced by angiotensin II. There appears to exist an important interdependence of these stimuli in the regulation of aldosterone secretion.


The mechanism of formation of renal stone crystals.
Typical renal stone minerals such as the calcium phosphates and oxalate hydrates may nucleate, grow, or dissolve in the fluctuating concentration conditions of the urinary tract. A study has been made of the rates of formation of these minerals under conditions in which the concentrations of ionic species were maintained constant by the potentiometrically controlled addition of lattice ions. The kinetic studies were made under conditions of low supersaturation similar to those in vivo and, in the case of calcium phosphates, at least three solid phases have been shown to participate in the overall reaction depending upon factors such as pH, nature and concentration of supporting medium, supersaturation, ionic strength, temperature, and the presence of inhibitors of crystallisation. For the calcium oxalates, the mineralisation process may involve the initial precipitation of less stable hydrates before conversion to the most stable whewellite. Urinary stone inhibitors may have a considerable influence in stabilising thermodynamically less stable hydrates.


Hepatolithiasis in East Asia. Retrospective study.
Hepatolithiasis is a major disease in Asia but differences in operative incidence between countries have not been examined. A retrospective study was conducted in Taiwan, Hong Kong, and Singapore, and the results were compared with those in Japan with the aim of defining factors involved in the etiology of the condition. In order to ensure uniformity of the data collected, the same form was used throughout the study and was completed by the same personnel after reviewing the patient's record and radiographs in each case. The years 1976-1980 were chosen for the study, since the newer methods of diagnosis such as ultrasound, endoscopic retrograde cholangiography, and percutaneous transhepatic cholangiography became available during that period. The most significant finding was the difference in the relative prevalence of hepatolithiasis as a proportion of all gallstone cases in Taiwan, Hong Kong, and Singapore, where the majority of the population consisted of patients of Chinese descent. The highest prevalence, 53.5%, was found in Taiwan, while in Hong Kong it was 3.1% and in Singapore 1.7%. Environmental rather than ethnic factors are implicated in the cause of hepatolithiasis.


Prophylactic effect of phenytoin in bipolar disorder: a controlled study.
Phenytoin is an effective anticonvulsant that has not previously been studied prophylactically in bipolar (BP) patients. Thus a study of phenytoin prophylaxis was undertaken and is herein reported. Bipolar patients were studied who had at least one episode per year in the previous 2 years despite ongoing prophylaxis. Patients were stable for a mean of 4 months (range 1-13) before entering the study. Phenytoin or placebo was added to their current therapy in a double-blind cross-over design for 6 months in each phase. Thirty observation periods of 6 months each were studied for 23 patients. Three patients had relapse on phenytoin and nine had relapse on placebo. There was a significant prophylactic effect of phenytoin in BP disorder [Cox's F-test for comparing survival in two groups: F(6, 18) = 3.44, p = 0.02]. This study suggests prophylactic effects of add-on phenytoin in BP illness. However, the number of patients was small and confirmation is necessary.


Influence of catheter position on estimated strain in intravascular elastography.
In elastography, an erroneous strain estimate is obtained when the radial strain and the probing ultrasound beam are not properly aligned: the "strain projection artifact". In practice, an angle between the strain and the ultrasound beam will be present in most of the cases due to inhomogeneities or nonuniform compression. In this study, a theoretical function describing the strain projection artifact is derived as a function of the angle between the radial strain and the ultrasound beam. Two main factors for an angle between strain and ultrasound beam in intravascular elastographic experiments are eccentricity and tilt of the transducer. The theoretical functions describing these errors are corroborated with strain estimates from an experiment with a circular, homogeneous gel-based vessel phantom. Comparison between the theoretical functions and the experimental results reveals that the strain projection artifact is well described by the theoretical findings. As a result, the experimental data can be corrected for this artifact. The corrected elastograms reveal that correct strain estimates are obtained when the eccentricity of the intravascular catheter is less than 63%. An "off-the-wall" device may be required to advance intravascular elastography to in vivo implementation.


Copper Transport and Disease: What Can We Learn from Organoids?
Many metals have biological functions and play important roles in human health. Copper (Cu) is an essential metal that supports normal cellular physiology. Significant research efforts have focused on identifying the molecules and pathways involved in dietary Cu uptake in the digestive tract. The lack of an adequate in vitro model for assessing Cu transport processes in the gut has led to contradictory data and gaps in our understanding of the mechanisms involved in dietary Cu acquisition. The recent development of organoid technology has provided a tractable model system for assessing the detailed mechanistic processes involved in Cu utilization and transport in the context of nutrition. Enteroid (intestinal epithelial organoid)-based studies have identified new links between intestinal Cu metabolism and dietary fat processing. Evidence for a metabolic coupling between the dietary uptake of Cu and uptake of fat (which were previously thought to be independent) is a new and exciting finding that highlights the utility of these three-dimensional primary culture systems. This review has three goals: (a) to critically discuss the roles of key Cu transport enzymes in dietary Cu uptake; (b) to assess the use, utility, and limitations of organoid technology in research into nutritional Cu transport and Cu-based diseases; and (c) to highlight emerging connections between nutritional Cu homeostasis and fat metabolism.


Extreme ultraviolet digital in-line holography using a tabletop source.
Digital in-line holography (DIH) offers fast, lensless, and aberration-free imaging with diffraction-limited resolution and inherently combines phase- and amplitude-contrast imaging, as well as three-dimensional imaging. Extending this technique to shorter wavelengths allows increasing the achievable spatial and phase-contrast resolution, as well as accessing material parameters not accessible in the optical domain. In this paper, we report on DIH experiments conducted with a coherent tabletop ultrafast high harmonic source operated at 38 nm wavelength. Applying a twin-image-free reconstruction scheme optimized for highly absorbing samples, we were able to demonstrate the phase-contrast imaging of silicon nitride sheets of 15 nm thickness and the use of the strong absorption of extreme ultraviolet in matter for amplitude-contrast imaging of thin films with spatial resolution below 1 μm. High-resolution morphology determination in combination with phase-contrast imaging is of special importance in thin-film characterization and applications arising thereof.


Iatrogenic meningitis: an increasing role for resistant viridans streptococci? Case report and review of the last 20 years.
Iatrogenic meningitis following lumbar puncture is a rare event. We present a 52-y-old man who developed symptoms of meningitis within 12 h after spinal anaesthesia. Cerebrospinal fluid cultures grew Streptococcus salivarius partially resistant to penicillin and ceftriaxone. The patient was successfully treated with ceftriaxone and vancomycin and left the hospital with minor sequelae. A literature review of 60 cases revealed the median age of the patients to be 44 y. The median incubation period was 24 h. Most cases occurred after spinal anaesthesia (n = 27), myelography (n = 20) and diagnostic lumbar puncture (n = 5). Organisms were isolated in 52 cases, and streptococcal species were responsible for 33 (63%) of them. An upward trend in resistance of S. viridans isolates is cause for concern and may change empirical treatment strategies. Death was reported in 3 cases (5%) and was associated with Pseudomonas and staphylococcal isolates. The recognition of this entity and the importance of proper infection control measures are underlined.


Cognitive Impairments in Schizophrenia as Assessed Through Activation and Connectivity Measures of Magnetoencephalography (MEG) Data.
The cognitive dysfunction present in patients with schizophrenia is thought to be driven in part by disorganized connections between higher-order cortical fields. Although studies utilizing electroencephalography (EEG), PET and fMRI have contributed significantly to our understanding of these mechanisms, magnetoencephalography (MEG) possesses great potential to answer long-standing questions linking brain interactions to cognitive operations in the disorder. Many experimental paradigms employed in EEG and fMRI are readily extendible to MEG and have expanded our understanding of the neurophysiological architecture present in schizophrenia. Source reconstruction techniques, such as adaptive spatial filtering, take advantage of the spatial localization abilities of MEG, allowing us to evaluate which specific structures contribute to atypical cognition in schizophrenia. Finally, both bivariate and multivariate functional connectivity metrics of MEG data are useful for understanding how these interactions in the brain are impaired in schizophrenia, and how cognitive and clinical outcomes are affected as a result. We also present here data from our own laboratory that illustrates how some of these novel functional connectivity measures, specifically imaginary coherence (IC), are quite powerful in relating disconnectivity in the brain to characteristic behavioral findings in the disorder.


Jahnula species from North and Central America, including three new species.
Three new species of loculoascomycetes collected from freshwater habitats in North America are described as new species of Jahnula (Jahnulales, Dothideomycetes). All three share these morphological features: hyaline to blackish translucent, membranous ascomata with subtending, wide, septate brown, spreading hyphae; peridia composed of large angular cells; hamathecium of septate pseudoparaphyses; 8-spored, clavate to cylindrical asci; and 1-septate, broadly fusiform, brown, multiguttulate ascospores. Four additional species, J. aquatica, J. bipolaris, J. potamophila, and J. seychellensis, are reported for the first time from the western hemisphere.


Interrupted Intramolecular Hydroaminomethylation of N-Protected-2-vinyl Anilines: Novel Access to 3-Substitued Indoles or Indoline-2-ols.
A new synthetic alternative to the synthesis of 3-methyl indoles and 3-methyl indoline-2-ols with an excellent atomic economy is presented in this study. It is demonstrated that the intramolecular interrupted hydroaminomethylation (HAM) reaction is a powerful tool for the formation of these compounds, which exhibit wide-ranging biological activity. Several N-Protected-2-vinyl anilines were synthesized and involved in the reaction producing the corresponding 3-methylindole or 3-methyl indoline-2-ol depending on the nature of the N-protecting groups.


Excess homocysteine upregulates the NRF2-antioxidant pathway in retinal Müller glial cells.
This study evaluated the effects of elevated homocysteine (Hcy) on the oxidative stress response in retinal Müller glial cells. Elevated Hcy has been implicated in retinal diseases including glaucoma and optic neuropathy, which are characterized by retinal ganglion cell (RGC) loss. To understand the mechanisms of Hcy-induced RGC loss, in vitro and in vivo models have been utilized. In vitro isolated RGCs are quite sensitive to elevated Hcy levels, while in vivo murine models of hyperhomocysteinemia (HHcy) demonstrate a more modest RGC loss (∼20%) over a period of many months. This differential response to Hcy between isolated cells and the intact retina suggests that the retinal milieu invokes mechanisms that buffer excess Hcy. Oxidative stress has been implicated as a mechanism of Hcy-induced neuron loss and NRF2 is a transcription factor that plays a major role in regulating cytoprotective responses to oxidative stress. In the present study we investigated whether HHcy upregulates NRF2-mediated stress responses in Müller cells, the chief retinal glial cell responsible for providing trophic support to retinal neurons. Primary Müller cells were exposed to L-Hcy-thiolactone [50μM-10mM] and assessed for viability, reactive oxygen species (ROS), and glutathione (GSH) levels. Gene/protein levels of Nrf2 and levels of NRF2-regulated antioxidants (NQO1, CAT, SOD2, HMOX1, GPX1) were assessed in Hcy-exposed Müller cells. Unlike isolated RGCs, isolated Müller cells are viable over a wide range of Hcy concentrations [50 μM - 1 mM]. Moreover, when exposed to elevated Hcy, Müller cells demonstrate decreased oxidative stress and decreased ROS levels. GSH levels increased by ∼20% within 24 h exposure to Hcy. Molecular analyses revealed 2-fold increase in Nrf2 expression. Expression of antioxidant genes Nqo1, Cat, Sod2, Hmox1, Gpx1 increased significantly. The consequences of Hcy exposure were evaluated also in Müller cells harvested from Nrf2-/- mice. In contrast to WT Müller cells, in which oxidative stress decreased upon exposure to Hcy, the Nrf2-/- Müller cells showed a significant increase in oxidative stress. Our data suggest that at least during early stages of Hhcy, a cytoprotective response may be in place, mediated in part by NRF2 in Müller cells.


Role of the intramolecular hydrogen bond network in the inhibitory power of chymotrypsin inhibitor 2.
A series of mutants of chymotrypsin inhibitor 2 (CI2), at residues involved in intramolecular interactions that shape and constrain the binding loop, were studied to determine their relative importance for inhibition of the serine protease subtilisin BPN', and for resistance of the inhibitor to proteolysis. These functional properties were investigated in tandem with the crystal structures of the mutant inhibitor-enzyme complexes. A dense hydrogen bonding network that supports the binding loop in the vicinity of the scissile bond was found to be important both for enzyme affinity and for stability to proteolysis. Structural analysis, in combination with biochemical measurements, allows differentiation of the structural components most important for resistance to proteolysis and/or binding. The most critical participating residues in the network were found to be Thr-58, Glu-60, Arg-65, and Gly-83. Glu-60 is more important for resistance to proteolysis than for binding, while Arg-65 and two other Arg residues play a greater role in binding than in resistance to proteolysis. Structural comparisons reveal a wide variety of subtle conformational changes in response to mutation, with built-in robustness in the hydrogen bond network, such that loss of one contact is compensated by other new contacts.


Long-acting bronchodilators and arterial stiffness in patients with COPD: a comparison of fluticasone furoate/vilanterol with tiotropium.
Increased arterial stiffness as measured by aortic pulse wave velocity (aPWV) predicts cardiovascular events and mortality and is elevated in patients with COPD. Prior investigation suggests that a long-acting β-agonist (LABA)/inhaled corticosteroid (ICS) lowers aPWV in patients with baseline aPWV ≥ 11 m/s. This study compared the effect of the ICS/LABA fluticasone furoate/vilanterol (FF/VI), 100/25 μg, delivered via the ELLIPTA dry powder inhaler, with tiotropium bromide (TIO), 18 μg, on aPWV. This multicenter, randomized, blinded, double-dummy, parallel-group, 12-week study compared FF/VI and TIO, both administered once daily. The primary end point was aPWV change from baseline at 12 weeks. Safety end points included adverse events (AEs), vital signs, and clinical laboratory tests. Two hundred fifty-seven patients with COPD and aPWV ≥ 11 m/s were randomized; 87% had prior cardiovascular events and/or risk. The mean difference in aPWV between FF/VI and TIO at week 12 was not significant (P = .484). Because the study did not contain a placebo arm, a post hoc analysis was performed to show that both treatments lowered aPWV by an approximate difference of 1 m/s compared with baseline. The proportion of patients reporting AEs was similar with FF/VI (24%) and TIO (18%). There were no changes in clinical concern for vital signs or clinical laboratory tests. No differences on aPWV were observed between FF/VI and TIO. However, further studies with a placebo arm are required to establish definitively whether long-acting bronchodilators lower aPWV. Both treatments demonstrated an acceptable tolerability profile. ClinicalTrials.gov; No.: NCT01395888; URL: www.clinicaltrials.gov.


Use of Surface Electromyography to Measure Muscle Fatigue in Patients in an Acute Care Hospital.
Functional outcomes with early rehabilitation in the acute care setting have improved; however, an improved understanding of muscle fatigue using surface electromyography (sEMG) is warranted to better guide patient-centered exercise prescription. The objectives of this study were to assess the safety and feasibility of collecting sEMG at the acute care bedside and to determine differences in muscle fatigue during isometric and dynamic submaximal contractions among patients in the hospital, healthy younger participants, and healthy older participants. The study used an observational cohort design. There were 37 participants. Median frequency (Fmed) of the myoelectric signal of the quadriceps femoris muscles and time to task failure (TTTF) were measured using sEMG during an isometric and dynamic fatiguing contraction. Primary analysis compared TTTF between groups for both types of contractions. Secondary analysis compared Fmed at initiation and termination of fatiguing contraction. High-quality sEMG measures were safe and feasible to collect at the acute care bedside with no adverse events. There was a statistically significant difference in TTTF between groups after isometric and dynamic contractions; hospitalized patients fatigued faster than healthy younger and healthy older participants after both contractions. With the exception of the vastus lateralis during a dynamic contraction in healthy younger and hospitalized patients, there was a statistically significant difference between Fmed at initiation and termination of contraction, indicating that subjects' muscles did truly fatigue. A limitation of the study was the small sample size of patients who were hospitalized without matched controls. sEMG is a lab quantitative technique that was found to be safe and feasible to assess muscle fatigue in the acute care environment. The protocol yielded similar results to previously published literature for healthy younger and healthy older people. Further research is needed to better understand how to integrate sEMG findings into patient-centered exercise prescriptions.


The effects of public education on the incidence and presentation of cutaneous melanoma in Central Texas.
Despite intensive public education, the incidence of melanoma in Central Texas has remained relatively constant (from 175 per 100,000 during 1981-1990 to 190 per 100,000 during 1991-2000). Although other investigators have hypothesized that the slight increase in incidence results from increased detection and reporting of early-stage (thinner) tumors, we found the average thickness was 0.99 mm from 1981 through 1990 and 1.11 mm from 1990 through 2000 (P = .30). These data suggest that despite efforts at public education, the incidence of melanoma in Central Texas is not decreasing and people are not identifying their tumors at an earlier stage.


Intrapulmonary airway smooth muscle is hyperreactive with a distinct proteome in asthma.
Constriction of airways during asthmatic exacerbation is the result of airway smooth muscle (ASM) contraction. Although it is generally accepted that ASM is hypercontractile in asthma, this has not been unambiguously demonstrated. Whether airway hyperresponsiveness (AHR) is the result of increased ASM mass alone or also increased contractile force generation per unit of muscle directly determines the potential avenues for treatment.To assess whether ASM is hypercontractile we performed a series of mechanics measurements on isolated ASM from intrapulmonary airways and trachealis from human lungs. We analysed the ASM and whole airway proteomes to verify if proteomic shifts contribute to changes in ASM properties.We report an increase in isolated ASM contractile stress and stiffness specific to asthmatic human intrapulmonary bronchi, the site of increased airway resistance in asthma. Other contractile parameters were not altered. Principal component analysis (PCA) of unbiased mass spectrometry data showed clear clustering of asthmatic subjects with respect to ASM specific proteins. The whole airway proteome showed upregulation of structural proteins. We did not find any evidence for a difference in the regulation of myosin activity in the asthmatic ASM.In conclusion, we showed that ASM is indeed hyperreactive at the level of intrapulmonary airways in asthma. We identified several proteins that are upregulated in asthma that could contribute to hyperreactivity. Our data also suggest enhanced force transmission associated with enrichment of structural proteins in the whole airway. These findings may lead to novel directions for treatment development in asthma.


Refractive surgery for the correction of myopia.
Interest in the surgical correction of myopia is increasing. At the present time, two procedures are clinically employed: radial keratotomy and myopic keratomileusis. The two techniques are described, and a detailed clinical comparison is presented.


The influence of changes in lipid values induced by cholestyramine and diet on progression of coronary artery disease: results of NHLBI Type II Coronary Intervention Study.
The National Heart, Lung and Blood Institute Type II Coronary Intervention Study, a double-blind, placebo-controlled trial, evaluated the efficacy of reduction in cholesterol levels induced by cholestyramine on progression of coronary artery disease (CAD). The rate of CAD progression in patients treated with cholestyramine plus diet was compared with that of patients treated with placebo plus diet. CAD progression was defined angiographically. Significant decrease in total cholesterol (TC) and low-density lipoprotein cholesterol (LDLc) and increases in high-density lipoprotein cholesterol (HDLc), as well as in HDLc/TC and HDLc/LDLc ratios, were observed with cholestyramine. HDLc change was due to increase in HDL2A and HDL2B. When the relationship between CAD progression and lipid changes was examined independent of specific treatment group, a significant inverse relationship was found between progression at 5 years and the combination of an increase in HDLc and a decrease in LDLc; changes in HDLc/TC and HDLc/LDLc were the best predictors of CAD change. While the testing of these relationships independent of treatment group was not part of the initial study design, the trends were observed in both the placebo-treated and cholestyramine-treated groups. Moreover, with multivariate analysis, the effect of cholestyramine treatment on CAD progression was eliminated by adding changes in HDLc/TC to the regression model. These findings support the hypothesis that increases in HDLc and decreases in TC (or LDLc) can prevent or delay CAD progression.


Health status among US Hispanics: ethnic variation, nativity, and language moderation.
The relationship between health status and Hispanic ethnicities, language, and nativity is poorly understood, due to the limitations and conflicting findings of previous studies. To examine the effects of language and nativity on health status in Hispanic ethnic subgroups and non-Hispanic whites (whites). Cross-sectional analyses of data from the 1998-2004 National Health Interview Survey linked to the 1999-2005 Medical Expenditure Panel Survey. Health status was regressed on race/ethnicity, interview language, and nativity, with adjustment for demographic and socioeconomic variables. A total of 16,489 Hispanics (13,522 Mexicans, 778 Cubans, 1360 Puerto Ricans, and 829 Dominicans) and 45,422 whites. SF-12 mental (MCS-12) and physical (PCS-12) component summary scores. In adjusted analyses, Mexicans had significantly higher MCS-12 scores than other Hispanics and whites, with the largest advantage noted for Spanish-speaking Mexicans. Ethnic origin * nativity interaction effects were significant for both MCS-12 [adjusted Wald test, F (3236) = 7.27, P = 0.0001] and PCS-12 [F (3236) = 4.75, P = 0.0031]. Continental US-born Mexicans had worse mental and physical health status than non-US-born Mexicans. By contrast, continental US birth was associated with better mental health status for Cubans and Dominicans, and better physical health status for Puerto Ricans. Complex interactions exist among language, nativity, ethnicity, and health status among Hispanics. Mexicans have better health status than whites and other Hispanics, and the moderating effects of nativity and language differ for Mexicans compared with other Hispanics. Future research should approach Hispanics as a diverse grouping rather than a monolithic entity.


Reduced effect of percutaneous retrogasserian glycerol rhizolysis in trigeminal neuralgia affecting the third branch.
The objective of this study was to determine the long-term efficacy of percutaneous glycerol rhizolysis of the trigeminal ganglion for treating patients with trigeminal neuralgia and search for predictors associated with (long-term) benefit to improve patient selection. A retrospective study in 60 consecutive patients treated with percutaneous glycerol rhizolysis of the trigeminal ganglion for trigeminal neuralgia. Charts were reviewed in combination with follow-up by questionnaire (n = 55, 92% response). Initial pain relief was achieved in 92% of the patients. Pain-free survival was 59% of the patients at 12 months and 53% at 24 months. Most common side effects were hypesthesia (15%), dry eye (5%), and meningitis (2%). In patients without involvement of the third branch of the trigeminal nerve, the initial effect was 79%, of which 90% achieved more than 2 years pain-free survival, compared with 97% initial effect and less than 40% pain-free survival of more than 2 years when the third trigeminal branch was involved. The present study demonstrates that involvement of the third branch is a negative predictor for long-term outcome in percutaneous glycerol rhizolysis of the trigeminal ganglion in patients with classical trigeminal neuralgia. However, in the absence of third-branch involvement, glycerol rhizolysis yields excellent long-term results when initial positive effect is obtained.


Packaging of prions into exosomes is associated with a novel pathway of PrP processing.
Prion diseases are fatal, transmissible neurodegenerative disorders associated with conversion of the host-encoded prion protein (PrP(C)) into an abnormal pathogenic isoform (PrP(Sc)). Following exposure to the infectious agent (PrP(Sc)) in acquired disease, infection is propagated in lymphoid tissues prior to neuroinvasion and spread within the central nervous system. The mechanism of prion dissemination is perplexing due to the lack of plausible PrP(Sc)-containing mobile cells that could account for prion spread between infected and uninfected tissues. Evidence exists to demonstrate that the culture media of prion-infected neuronal cells contain PrP(Sc) and infectivity but the nature of the infectivity remains unknown. In this study we have identified PrP(C) and PrP(Sc) in association with endogenously expressing PrP neuronal cell-derived exosomes. The exosomes from our prion-infected neuronal cell line were efficient initiators of prion propagation in uninfected recipient cells and to non-neuronal cells. Moreover, our neuronal cell line was susceptible to infection by non-neuronal cell-derived exosome PrP(Sc). Importantly, these exosomes produced prion disease when inoculated into mice. Exosome-associated PrP is packaged via a novel processing pathway that involves the N-terminal modification of PrP and selection of distinct PrP glycoforms for incorporation into these vesicles. These data extend our understanding of the relationship between PrP and exosomes by showing that exosomes can establish infection in both neighbouring and distant cell types and highlight the potential contribution of differentially processed forms of PrP in disease distribution. These data suggest that exosomes represent a potent pool of prion infectivity and provide a mechanism for studying prion spread and PrP processing in cells endogenously expressing PrP.


A birth cohort study of viral infections in Vietnamese infants and children: study design, methods and characteristics of the cohort.
In Ho Chi Minh City, Vietnam, more than one-third of admissions to the two paediatric hospitals are attributable to four infectious syndromes: dengue, diarrhoeal disease, acute respiratory infection, and hand, foot and mouth disease. We have established a large prospective birth cohort study to investigate individual, environmental, virological, and immunological determinants of infection and disease in infants. Specific research questions are focused on the role of maternal antibody in protection against infection in infancy, and the adaptive immune response to vaccination and natural infection. This paper presents the cohort design, methods, and baseline characteristics of the participants enrolled in the first two years. Women are enrolled prior to delivery at one hospital in each of two catchment areas: an urban district in central HCMC, and a mixed urban/rural district in the Mekong Delta 150 km southwest of HCMC. Infants are enrolled within 3 days of birth, and maternal and cord blood samples are collected. Routine blood samples and data on growth, health status and vaccinations are collected from infants at scheduled visits at 4, 9 and 12 months. Clinical data and specimens are collected from infants presenting at a study clinic, or admitted to hospital, with any of the the four infectious syndromes of interest. In four years since since the study began in July 2009, >6400 infants have been enrolled, and enrolment is ongoing. Attrition is low: 84% of participants have completed the full 12-month follow-up period. Baseline characteristics of the first 4300 enrollees are presented here. We have demonstrated the feasibility of establishing a large prospective study of infectious diseases in infancy in a resource-limited setting, with minimal loss to follow-up. Our linked socio-demographic, clinical and laboratory data will help elucidate the viral aetiology and epidemiology of common infectious diseases of infancy, and can inform the implemention of existing and future vaccines. This study furthermore provides a platform to which additional endpoints could be added in the future.


Exploring the gating mechanisms of aquaporin-3: new clues for the design of inhibitors?
The pH gating of human AQP3 and its effects on both water and glycerol permeabilities have been fully characterized for the first time using a human red blood cell model (hRBC). For comparison, the effects of pH on the gating of rat AQP3 have also been characterized in yeast. The obtained results highlight similarities as well as differences between the two isoforms. In addition, we investigated the molecular mechanism of hAQP3 pH gating in silico, which may disclose new pathways to AQP regulation by small molecule inhibitors, and therefore may be important for drug development.


Antimicrobial resistance and investigation of the molecular epidemiology of Listeria monocytogenes in dairy products.
Listeria monocytogenes is a ubiquitous microorganism in nature and is responsible for listeriosis, an infectious disease caused by consumption of contaminated food. Molecular characterization was performed on 19 strains of Listeria monocytogenes (serovars 1/2a, 1/2b, 4b and 4c), isolated from dairy products in Rio Grande do Sul, Brazil. The molecular techniques applied were random amplification of polymorphic DNA and restriction enzyme analysis. In addition to the molecular analysis, the antimicrobial resistance profile was determined. The strains studied showed a low degree of diversity. In relation to the antimicrobial resistance profile of those microorganisms from the samples analyzed, all of them were susceptible to the antimicrobials tested. The molecular techniques that were used presented good discriminatory power for the strains studied. Furthermore, all of the samples that were analyzed were susceptible to the antimicrobials tested.


A Longitudinal Examination of Decisions to Ride and Decline Rides with Drinking Drivers.
Riding with a drinking driver (RWDD) is a serious concern that leads to numerous preventable deaths every year. There is a significant gap in research on empirically tested predictors of RWDD that could be implemented in prevention efforts. College students are in need of such prevention efforts, as they have some of the highest rates of alcohol-related crash fatalities and may engage in RWDD more than their noncollege peers. This study utilized behavioral decision-making approach to examine predictors of RWDD and declining a ride from a drinking driver (Decline) in older college students. Students (n = 791) in their third year of college were enrolled from 3 large and diverse universities. Psychosocial (e.g., expectancies, norms) and decision-making variables (willingness to RWDD and intentions to use alternatives) were assessed in the fall of their third year. One year later, RWDD and Decline behaviors were assessed. Zero-inflated Poisson analyses were used to assess how decision-making variables predicted RWDD and Decline behavior. Associations between psychosocial and decision-making variables were also assessed. Thirteen percent of students reported RWDD and ~28% reported Decline behavior. Willingness to RWDD and typical weekly drinking were both associated with increases in RWDD (odds ratio [OR] = 1.58 and 1.40, respectively), whereas intentions to use alternatives, sex, and ethnicity were not associated with RWDD. Only weekly drinking was associated with Decline, with an increase in drinking associated with increased Decline (OR = 1.48). All psychosocial variables were significantly associated with the decision-making variables except positive expectancies. Results provide evidence that willingness to RWDD is a predictor of future RWDD, even if students intend to use safe alternatives. Future research is needed to better understand decision-making factors that influence Decline. Results also suggest prevention and interventions efforts, such as brief motivational intervention, Parent-Based Interventions, and normative feedback interventions could be adapted to reduce RWDD.


Value of histopathologic analysis of skin excisions by GPs.
The clinical diagnoses of skin lesions in general practice may sometimes not be very accurate. The aim of this study was to compare clinical versus final histopathological diagnosis status (benign, pre-malignant/malignant) in 4595 consecutive submissions by GPs. The final diagnosis was pre-malignant or malignant in 215 cases (4.7%). From the 4436 lesions clinically diagnosed as benign, 134 (3.0%) were pre-malignant or malignant on final histological diagnosis. From the 159 lesions clinically diagnosed as pre-malignant or malignant, 78 (49.1%) were in fact benign, and 81 (50.9%) were indeed pre-malignant or malignant on final diagnosis. The sensitivity for a malignant diagnosis was 38%, and the specificity 98%. The proportion of pre-malignancies or malignancies was 0.9% below and 9.2% above the age of 40 years. In conclusion, histopathological investigation of skin excisions by GPs yields a high percentage of unexpected pre-malignancies and malignancies. The number of misdiagnoses was age dependent, with a proportion of 1% and 9% of pre-malignancies/malignancies in patients below and above the age of 40 years, respectively. This indicates that all skin excisions by GPs must undergo routine histopathological investigation to ensure that serious malignancies are not missed.


Brassicaceae express multiple isoforms of biotin carboxyl carrier protein in a tissue-specific manner.
Plastidial acetyl-coenzyme A carboxylase from most plants is a multi-enzyme complex comprised of four different subunits. One of these subunits, the biotin carboxyl carrier protein (BCCP), was previously proposed to be encoded by a single gene in Arabidopsis. We report and characterize here a second Arabidopsis BCCP (AtBCCP2) cDNA with 42% amino acid identity to AtBCCP1 and 75% identity to a class of oilseed rape (Brassica napus) BCCPs. Both Arabidopsis BCCP isoforms were expressed in Escherichia coli and found to be biotinylated and supported carboxylation activity when reconstituted with purified, recombinant Arabidopsis biotin carboxylase. In vitro translated AtBCCP2 was competent for import into pea (Pisum sativum) chloroplasts and processed to a 25-kD polypeptide. Extracts of Arabidopsis seeds contained biotinylated polypeptides of 35 and 25 kD, in agreement with the masses of recombinant AtBCCP1 and 2, respectively. AtBCCP1 protein was present in developing tissues from roots, leaves, flowers, siliques, and seeds, whereas AtBCCP2 protein was primarily expressed in 7 to 10 d-after-flowering seeds at levels approximately 2-fold less abundant than AtBCCP1. AtBCCP1 transcript reflected these protein expression profiles present in all developing organs and highest in 14-d leaves and siliques, whereas AtBCCP2 transcript was present in flowers and siliques. In protein blots, four different BCCP isoforms were detected in developing seeds from oilseed rape. Of these, a 35-kD BCCP was detected in immature leaves and developing seeds, whereas developing seeds also contained 22-, 25-, and 37-kD isoforms highly expressed 21 d after flowering. These data indicate that oilseed plants in the family Brassicaceae contain at least one to three seed-up-regulated BCCP isoforms, depending upon genome complexity.


Endosulfan-induced hepatotoxicity is route of exposure independent in rats.
Endosulfan is an important hepatotoxic agent that generates free oxygen radicals in liver. With the widespread use of endosulfan in agriculture, human beings are most likely to be exposed to it by eating food contaminated with endosulfan, exposure to its low levels by skin contact with contaminated soil, smoking cigarettes made from tobacco that has endosulfan residues on it, or by nose and whole body inhalation exposure in the farms during its application. Since endosulfan is a frequently used pesticide, and the incidence of toxic injury to the liver tissue in relation to its widespread use reported in the literature, we considered it necessary to investigate whether endosulfan-induced liver injury could be route of exposure dependent. Eighteen mature male albino Wistar rats, weighing between 180 and 220 g, were used in this study. The hepatotoxic effects of oral administration of endosulfan (5 mg/kg body weight) daily for 30 days, and 30 days whole body inhalation exposure to ungraded concentration of endosulfan were investigated in rats using serum liver enzymes and histopathological assay. At the end of the experimental period, serum alanine aminotransferase, aspartate amino transferase, alkaline phosphatase, and creatine kinase activities obtained for the group of rats exposed orally to endosulfan were not significantly different (p ≥ 0.05) from the activities obtained for rats exposed by whole body inhalation. However, the activity of these enzymes obtained for the rats exposed to endosulfan by both oral and inhalation routes were significantly increased (p ≤ 0.05) compared, respectively, to the control. Also, on microscopic examination, the liver tissues of experimental groups exhibited severe damage histopathologically. The results of the enzyme and histological analyses showed that both oral and whole body inhalation exposure to endosulfan may cause liver tissue damage in rats. The exposure to endosulfan in rats caused liver tissue damage independent of the route of exposure.


Isolated acute non-cystic white matter injury in term infants presenting with neonatal encephalopathy.
We discuss possible aetiological factors, MRI evolution of injury and neuro-developmental outcomes of neonatal encephalopathy (NE). Thirty-six consecutive infants diagnosed with NE were included. In this cohort, four infants (11%) were identified with injury predominantly in the deep white matter on MRI who were significantly of younger gestation, lower birthweight with higher Apgars at one and five minutes compared to controls. Placental high grade villitis of unknown aetiology (VUA) was identified in all four of these infants. Our hypothesis states VUA may induce white matter injury by causing a local inflammatory response and/or oxidative stress during the perinatal period. We underline the importance of continued close and systematic evaluation of all cases of NE, including examination of the placenta, in order to come to a better understanding of the clinical presentation, the patterns of brain injury and the underlying pathophysiological processes.


Cavernous hemangioma: unusual benign tumor of the transverse colon.
Cavernous hemangioma of the colon is an uncommon disease and a rare cause of bleeding. Few cases have been reported in the literature. The rectosigmoid is the most common site of this disease in the gastrointestinal tract, while colonic localization is very uncommon. We report the case of a 66-year-old female with recurrent episodes of rectal bleeding. She underwent a colonoscopy and was diagnosed with diffuse cavernous hemangioma of the transverse colon. A laparoscopic extended right hemicolectomy was performed. Colonic hemangiomas are very rare vascular malformations and their clinical presentation is usually acute, recurrent or chronic rectal bleeding. This tumor can be diagnosed as solitary, multiple, or part of a more complex syndrome with cutaneous manifestations. They can also invade adjacent structures. Colonoscopy is a useful diagnostic test. The extension of the lesion, its morphology and its localization can be established, but imaging such as magnetic resonance or computerized tomography scan has to be performed. Sometimes, however, recognition of these tumors is difficult and can be a cause of failed surgical treatment and severe complications. Surgical treatment is recommended, after a full diagnostic evaluation for other causes of gastrointestinal bleeding.


Competitive reaction pathways in vibrationally induced photodissociation of H2SO4.
Vibrationally induced photodissociation of sulfuric acid into H2O + SO3 is investigated based on reactive molecular dynamics (MD) simulations. Multisurface adiabatic reactive MD simulations allow us to follow both, H-transfer and water elimination after excitation of the ν9 OH-stretching mode. Analysis of several thousand trajectories finds that the H2O and SO3 fragments have distinct final state distributions with respect to translational, rotational, and vibrational degrees of freedom. Rotational distributions peak at quantum numbers j ≤ 5 for water and j ≈ 60 for SO3. The final state distributions should be useful in identifying products in forthcoming experiments. Based on the MD trajectories, a kinetic scheme has been developed which is able to explain most of the trajectory data and suggests that IVR is very rapid. Typical lifetimes of the excited complex range from several 10 picoseconds to hundreds of nanoseconds, depending on the excitation level. Including temperature and pressure profiles characteristic for the stratosphere in the kinetic model shows that excitations higher than ν9 = 4 can significantly contribute to the photolysis rate. This extends and specifies earlier work in that multi-level modeling is required to understand the significance of vibrationally induced decomposition pathways of sulfuric acid in the middle atmosphere.


Does Candida albicans Als5p amyloid play a role in commensalism in Caenorhabditis elegans?
Candida albicans, a dimorphic fungus and an opportunistic pathogen, possesses a myriad of adherence factors, including members of the agglutinin-like sequence (Als) family of mannoproteins. The adhesin Als5p mediates adhesion to many substrates and is upregulated during commensal interactions but is downregulated during active C. albicans infections. An amyloid-forming core sequence at residues 325 to 331 is important for Als5p function, because a single-amino-acid substitution at position 326 (V326N) greatly reduces Als5p-mediated adherence. We evaluated the role of Als5p in host-microbe interactions by using Caenorhabditis elegans nematodes as a host model and feeding them Saccharomyces cerevisiae expressing Als5p on the surface. Als5p-expressing yeast had 8.5- and 3.5-fold-increased intestinal accumulation rates compared to Als5p-nonexpressing S. cerevisiae or yeast expressing amyloid-deficient Als5p(V326N), respectively. Surprisingly, this accumulation delayed S. cerevisiae-induced killing of C. elegans. The median survival time was nearly twice as long as that of nematodes fed nonexpressing or non-amyloid-forming Als5p(V326N)-expressing S. cerevisiae. Treatment with the amyloid-inhibiting dye Congo red or repression of Als5p expression abrogated the protective effect of Als5p. Furthermore, Als5p had no effect on oocyte quantity or quality, since nematodes fed either empty vector (EV)- or Als5p(V326N)-expressing S. cerevisiae had similar egg-laying and egg-hatching rates. This study is the first, to our knowledge, to show that expression of an amyloid-forming protein can attenuate pathogenicity in C. elegans.


Cooling and heating degree days in the US: The role of macroeconomic variables and its impact on environmental sustainability.
Beyond employing the cooling and heating degree days variables for evaluating the climatic conditions and the expected energy needs in the United States, a subtle concern is the underpinning role of the environmental sustainability amidst socio-economic activities. As such, the current study is design to examine the role of fossil fuel energy consumption, ecological footprint, and urban population on the degree days viz-a-vis the cooling and heating days in the United States over the period 1960-2015. The Autoregressive Distributed Lag bound testing model employed reveals the importance of the ecological footprint, fossil fuel energy consumption, and urban population on the cooling and heating degree days of the United States. Result posits that each of fossil fuel and the urban population plays a positive and negative role as regard the cooling degree days and the heating degree days respectively, especially in the long run. Importantly, the empirical results support the argument that the increase in the consumption of the fossil fuel sources of energies are responsible to cause more cooling degree days, thus resulting to longer and hotter periods in the United States but vice versa for the heating degree days. Similarly, the investigation outcome draws from the argument that the increase in the urban population is a potential cause of high environmental temperature, thus responsible for lengthy heat periods (cooling degree days) and resulting in more energy needs and technologies for cooling. As expected, the reverse is the case for the heating degree days, especially in the long-run. As a policy standpoint, policymakers are to further adopt improved and effective guideline for housing and building constructions that are weather specifics. In formulating policy vehicle for each of the seasonal dynamics, the economic benefits of each of the climatic measurements should be considered especially for both the short- and long-run environmental sustainability.


Effects of a Self-efficacy Theory-Based Training Program for Peers of Patients with Type 2 Diabetes.
Training peer leaders to deliver patient education is expected to be a low-cost approach to providing healthcare in urban-poor areas affected by a shortage of healthcare professionals. The purpose of this study was to examine the effects of a training program on the self-efficacy and knowledge of peer leaders with type 2 diabetes. A single-group longitudinal survey with baseline, intervention, and follow-up periods was conducted at a diabetes clinic in a small municipality in Metro Manila, Philippines. The intervention, a self-efficacy theory-based training program for peer-leaders of diabetic patients conducted in August 2017, comprised hands-on learning, demonstrations, quizzes, role-playing, group sharing, physical exercise, and a buffet lunch. The primary outcome was participants' self-efficacy for management of their diabetes. Secondary outcomes were participants' knowledge of diabetes and levels of emotional distress, motivation, and confidence for guiding their peers, satisfaction with the training program, hemoglobin A1c, and quality of life. At 12 and 18 months after the intervention, participants' knowledge of diabetes was significantly increased compared with baseline (both P < 0.05). At earlier time points, an increasing, but not significant, trend was observed. The change in knowledge of diabetes from baseline to 18 months after intervention tended to be positively correlated with the change in self-efficacy (r = 0.594, P = 0.054). No significant differences were observed for any of the other outcomes, although the descriptive statistics showed an increasing trend for all of the outcomes except motivation. The training program significantly improved participants' knowledge of diabetes at 12 and 18 months after the training programs compared with baseline. A positive correlation between the changes in the levels of knowledge and self-efficacy suggested that the observed improvement of self-efficacy was facilitated by the improvement of knowledge of diabetes.


Uncovering disassortativity in large scale-free networks.
Mixing patterns in large self-organizing networks, such as the Internet, the World Wide Web, and social and biological networks, are often characterized by degree-degree dependencies between neighboring nodes. In this paper, we propose a new way of measuring degree-degree dependencies. One of the problems with the commonly used assortativity coefficient is that in disassortative networks its magnitude decreases with the network size. We mathematically explain this phenomenon and validate the results on synthetic graphs and real-world network data. As an alternative, we suggest to use rank correlation measures such as Spearman's ρ. Our experiments convincingly show that Spearman's ρ produces consistent values in graphs of different sizes but similar structure, and it is able to reveal strong (positive or negative) dependencies in large graphs. In particular, we discover much stronger negative degree-degree dependencies in Web graphs than was previously thought. Rank correlations allow us to compare the assortativity of networks of different sizes, which is impossible with the assortativity coefficient due to its genuine dependence on the network size. We conclude that rank correlations provide a suitable and informative method for uncovering network mixing patterns.


Granulin-epithelin precursor overexpression promotes growth and invasion of hepatocellular carcinoma.
Granulin-epithelin precursor (GEP) is a novel growth factor. Our earlier cDNA microarray study indicated that GEP was overexpressed in hepatocellular carcinoma (HCC). The aim of this study was to investigate the clinical significance of GEP expression and its potential as a therapeutic target in HCC. A total of 110 pairs of HCCs and adjacent nontumor liver tissues, and 22 normal liver tissues were examined. The GEP RNA level was examined by quantitative reverse transcription-PCR, and protein localization by immunohistochemistry. The GEP function was examined by transfection experiments. The RNA levels of the HCCs were significantly higher than those of the nontumor liver tissues and normal livers (P <0.001). GEP protein staining was observed in tumor cytoplasm, and the GEP protein levels of the HCCs were also significantly higher than those of the nontumor liver tissues and normal livers (P <0.001). The majority of HCCs demonstrated up-regulation of GEP protein compared with their adjacent liver tissues [79 (71.8%) of 110]. Positive correlation of GEP RNA with protein levels was observed in HCCs (P <0.01). Strong GEP expression was associated with large HCCs, venous infiltration, and early intrahepatic recurrence (P <0.05). Functional studies on the HCC cell line Hep3B demonstrated that reduction of GEP protein levels resulted in decreased cell proliferation rates, tumor invasion ability, anchorage-independent growth in soft agar, and tumorigenicity in nude mice (P <0.05). GEP is an important factor for HCC growth, invasion, and metastasis. GEP has the potential to serve as a tumor marker and therapeutic target.


Reversible Charge Trapping in Bis-Carbazole-Diimide Redox Polymers with Complete Luminescence Quenching Enabling Nondestructive Read-Out by Resonance Raman Spectroscopy.
The coupling of substituted carbazole compounds through carbon-carbon bond formation upon one-electron oxidation is shown to be a highly versatile approach to the formation of redox polymer films. Although the polymerization of single carbazole units has been proposed earlier, we show that by tethering pairs of carbazoles double sequential dimerization allows for facile formation of redox polymer films with fine control over film thickness. We show that the design of the monomers and in particular the bridging units is key to polymer formation, with the diaminobenzene motif proving advantageous, in terms of the matching to the redox potentials of the monomer and polymer film and thereby avoiding limitations in film thickness (autoinsulation), but introduces unacceptable instability due to the intrinsic redox activity of this moiety. The use of a diimide protecting group both avoids complications due to p-diamino-benzene redox chemistry and provides for a redox polymer in which the photoluminescence of the bis-carbazole moiety can be switched reversibly (on/off) with redox control. The monomer design approach is versatile enabling facile incorporation of additional functional units, such as naphthalene. Here we show that a multicomponent carbazole/naphthalene containing monomer (APCNDI) can form redox polymer films showing both p- and n- conductivity under ambient conditions and allows access to five distinct redox states, and a complex electrochromic response covering the whole of the UV/vis-NIR spectral region. The highly effective quenching of the photoluminescence of both components in poly-APCNDI enables detailed characterization of the redox polymer films. The poly-APCNDI films show extensive charge trapping, which can be read out spectroscopically in the case of films and is characterized as kinetic rather than chemical in origin on the basis of UV/vis-NIR absorption and resonance Raman spectroscopic analyses. The strong resonantly enhanced Raman scattering for the various oxidized and reduced states of APCNDI enables nondestructive "read-out" of the state of the polymer, including that in which charges are trapped kinetically at the surface, making poly-APCNDI highly suitable for application as a component in organic nonvolatile memory devices.


L1 interaction domains of papillomavirus l2 necessary for viral genome encapsidation.
BPHE-1 cells, which harbor 50 to 200 viral episomes, encapsidate viral genome and generate infectious bovine papillomavirus type 1 (BPV1) upon coexpression of capsid proteins L1 and L2 of BPV1, but not coexpression of BPV1 L1 and human papillomavirus type 16 (HPV16) L2. BPV1 L2 bound in vitro via its C-terminal 85 residues to purified L1 capsomers, but not with intact L1 virus-like particles in vitro. However, when the efficiency of BPV1 L1 coimmunoprecipitation with a series of BPV1 L2 deletion mutants was examined in vivo, the results suggested that residues 129 to 246 and 384 to 460 contain independent L1 interaction domains. An L2 mutant lacking the C-terminal L1 interaction domain was impaired for encapsidation of the viral genome. Coexpression of BPV1 L1 and a chimeric L2 protein composed of HPV16 L2 residues 1 to 98 fused to BPV1 L2 residues 99 to 469 generated infectious virions. However, inefficient encapsidation was seen when L1 was coexpressed with either BPV1 L2 with residues 91 to 246 deleted or with BPV1 L2 with residues 1 to 225 replaced with HPV16 L2. Impaired genome encapsidation did not correlate closely with impairment of the L2 proteins either to localize to promyelocytic leukemia oncogenic domains (PODs) or to induce localization of L1 or E2 to PODs. We conclude that the L1-binding domain located near the C terminus of L2 may bind L1 prior to completion of capsid assembly, and that both L1-binding domains of L2 are required for efficient encapsidation of the viral genome.


Hospitalization risk of the 2009 H1N1 pandemic cases in Hong Kong.
Reliable assessment for the severity of the 2009 H1N1 pandemic influenza is critical for evaluation of vaccination strategies for future pandemics. This study aims to estimate the age-specific hospitalization risks of the 2009 pandemic cases during the first wave in Hong Kong, by combining the findings from the serology and disease burden studies. Excess hospitalization rates associated with the pandemic H1N1 were estimated from Poisson regression models fitted to weekly total numbers of non-accidental hospitalization from 2005 to 2010. Age-specific infection-hospitalization risks were calculated as excess hospitalization rates divided by the attack rates in the corresponding age group, which were estimated from serology studies previously conducted in Hong Kong. Excess hospitalization rate associated with pandemic H1N1 was highest in the 0-4 age group (881.3 per 100,000 population), followed by the 5-14, 60+, 15-29, 50-59, 30-39 and 40-49 age groups. The hospitalization risk of the infected cases (i.e. infection-hospitalization risk) was found highest in the 60+ age group and lowest in the 15-29 age group, with the estimates of 17.5% and 0.7%, respectively. People aged 60 or over had a relatively high infection-hospitalization risk during the first wave of the 2009 H1N1 pandemic, despite of a low attack rate in this age group. The findings support the policy of listing older people as the priority group for pandemic vaccination.