Mining and analysis of audiology data to find significant factors associated with tinnitus masker.
The objective of this research is to find the factors associated with tinnitus masker from the literature, and by using the large amount of audiology data available from a large NHS (National Health Services, UK) hearing aid clinic. The factors evaluated were hearing impairment, age, gender, hearing aid type, mould and clinical comments. The research includes literature survey for factors associated with tinnitus masker, and performs the analysis of audiology data using statistical and data mining techniques. This research uses a large audiology data but it also faced the problem of limited data for tinnitus. It uses 1,316 records for tinnitus and other diagnoses, and 10,437 records of clinical comments from a hearing aid clinic. The research is looking for variables associated with tinnitus masker, and in future, these variables can be combined into a single model to develop a decision support system to predict about tinnitus masker for a patient. The results demonstrated that tinnitus maskers are more likely to be fit to individuals with milder forms of hearing loss, and the factors age, gender, type of hearing aid and mould were all found significantly associated with tinnitus masker. In particular, those patients having Age < =55 years were more likely to wear a tinnitus masker, as well as those with milder forms of hearing loss. ITE (in the ear) hearing aids were also found associated with tinnitus masker. A feedback on the results of association of mould with tinnitus masker from a professional audiologist of a large NHS (National Health Services, UK) was also taken to better understand them. The results were obtained with different accuracy for different techniques. For example, the chi-squared test results were obtained with 95% accuracy, for Support and Confidence only those results were retained which had more than 1% Support and 80% Confidence. The variables audiograms, age, gender, hearing aid type and mould were found associated with the choice of tinnitus masker in the literature and by using statistical and data mining techniques. The further work in this research would lead to the development of a decision support system for tinnitus masker with an explanation that how that decision was obtained.


Neuropsychological studies in anorexia nervosa.
Neuropsychological findings in eating disorders are somewhat inconsistent. This may be because individual studies have used a broad range of tests on relatively small, heterogeneous clinical groups, thus limiting the detection of subtle neuropsychological differences in these patients. Therefore, rather than using broad assessments of a variety of neuropsychological functions, adoption of a more focused, hypothesis-driven approach based on clinical practice is proposed. This will allow more in-depth investigations of targeted functions and will improve the chances of detecting a problem, of exploring its ecologic validity, and of tailoring a treatment. We have demonstrated this approach using our neuropsychological studies of cognitive flexibility in anorexia nervosa (AN).


Smoking cessation, but not reduction, reduces cardiovascular disease incidence.
The aim of this study was to assess the association of smoking cessation and reduction with risk of cardiovascular disease (CVD). A total of 897 975 current smokers aged ≥40 years who had undergone two consecutive national health examinations (in 2009 and 2011) were included. Participants were classified as quitters (20.6%), reducers I (≥50% reduction, 7.3%), reducers II (20-50% reduction, 11.6%), sustainers (45.7%), and increasers (≥20% increase, 14.5%). During 5 575 556 person-years (PY) of follow-up, 17 748 stroke (3.2/1000 PY) and 11 271 myocardial infarction (MI) (2.0/1000 PY) events were identified. Quitters had significantly decreased risk of stroke [adjusted hazard ratio (aHR) 0.77 95% confidence interval (CI) 0.74-0.81; absolute risk reduction (ARR) -0.37, 95% CI -0.43 to -0.31] and MI (aHR 0.74, 95% CI 0.70-0.78; ARR -0.27, 95% CI -0.31 to -0.22) compared to sustainers after adjustment for demographic factors, comorbidities, and smoking status. The risk of stroke and MI incidence in reducers I (aHR 1.02, 95% CI 0.97-1.08 and aHR 0.99, 95% CI 0.92-1.06, respectively) and reducers II (aHR 1.00, 95% CI 0.95-1.05 and aHR 0.97, 95% CI 0.92-1.04, respectively) was not significantly different from the risk in sustainers. Further analysis with a subgroup who underwent a third examination (in 2013) showed that those who quit at the second examination but had starting smoking again by the third examination had 42-69% increased risk of CVD compared to sustained quitters. Smoking cessation, but not reduction, was associated with reduced CVD risk. Our study emphasizes the importance of sustained quitting in terms of CVD risk reduction.


Three classes of inhibitory amino acid terminals in the cochlear nucleus of the guinea pig.
Electron microscopic postembedding immunocytochemistry was used to analyze and assess the synaptic distribution of glycine (GLY) and gamma-amino butyric acid (GABA) immunoreactivities in the guinea pig cochlear nucleus (CN). Three classes of endings were identified containing immunolabeling for glycine, GABA, or both glycine and GABA (GLY/GABA). All classes were similar in that the terminals contained pleomorphic vesicles and formed symmetric synapses with their postsynaptic targets. A fourth class, which labeled with neither antibody, contained round vesicles and formed asymmetric synapses. Glycine endings predominated in the ventral CN, while GLY/GABA endings were prevalent in the dorsal CN. GABA endings were the least common and smallest in size. Glycine, GLY/GABA, and GABA endings differed in their proportions and patterns of distribution on the different classes of projection neurons in the CN, including spherical bushy, type I stellate/multipolar, and octopus cells in the ventral CN and fusiform cells in the dorsal CN. The vast majority of anatomically-defined, putative inhibitory endings contain GLY, GABA, or both, suggesting that most of the inhibition in the cochlear nucleus is mediated by these three cytochemically and, probably, functionally distinct classes of endings. The results of this study also suggest that a large proportion of the GABA available for inhibition in the CN coexists in terminals with glycine.


Watson will see you now: a supercomputer to help clinicians make informed treatment decisions.
IBM has collaborated with several cancer care providers to develop and train the IBM supercomputer Watson to help clinicians make informed treatment decisions. When a patient is seen in clinic, the oncologist can input all of the clinical information into the computer system. Watson will then review all of the data and recommend treatment options based on the latest evidence and guidelines. Once the oncologist makes the treatment decision, this information can be sent directly to the insurance company for approval. Watson has the ability to standardize care and accelerate the approval process, a benefit to the healthcare provider and the patient.


Gene expression profiles of ileal inflammatory bowel disease correlate with disease phenotype and advance understanding of its immunopathogenesis.
Pouchitis may develop in patients with ulcerative colitis undergoing pouch surgery. We aimed to evaluate the de novo inflammation developing in the ileal pouch, hypothesizing that it may be similar to ileitis in Crohn's disease (CD). Patients with ulcerative colitis pouch were prospectively recruited, stratified according to disease behavior into normal pouch, chronic pouchitis, and Crohn's-like disease of the pouch groups, and compared with controls. Gene expression analysis was performed using microarrays, validated by real-time polymerase chain reaction. Gene ontology and clustering were evaluated using bioinformatic tools. Sixty-six subjects were recruited. Although in ulcerative colitis ileum there were no significant gene expression alterations, patients with normal pouch had 168 significant alterations (fold change ≥ 2, corrected P ≤ 0.05). In chronic pouchitis and Crohn's-like disease of the pouch, 490 and 1152 alterations were detected, respectively. High degree of overlap in gene expression alterations between the pouch subgroups was demonstrated. The magnitude of change correlated with pouch disease behavior. Gene expression profiles were more reflective of disease behavior compared with inflammatory indices. CD ileitis had 358 alterations, with a 90% overlap with pouchitis. Gene ontology analyses revealed multiple biological processes associated with pouch inflammation, including response to chemical stimulus, small molecule metabolic and immune system processes, and specific infection-related pathways such as Staphylococcus aureus, leishmaniasis, and tuberculosis. Gene alterations in pouch inflammation and CD overlap, suggesting that inflammatory bowel diseases is a spectrum, rather than distinct diseases. Pouchitis may serve as a model of CD. The novel pathways associated with inflammatory bowel diseases may decipher pathophysiology and suggest targets for intervention.


[Gestational malaria: HELLP syndrome mistaken diagnosis].
Malaria is one of the most important parasitic infections in Mexico and Latin America. Here we report a case of a 21 year-old female with 38.4 weeks of pregnancy and previous hospitalization due to malaria. Showing a thick drop negative test she was referred to Mexico City Hospital de la Mujer with presumptive diagnosis of preeclampsia and HELLP syndrome. During her stay in ICU she developed malarial paroxysm and Plasmodium vivax was identified, conducting to specific therapy.


Himalayan Saccharomyces eubayanus Genome Sequences Reveal Genetic Markers Explaining Heterotic Maltotriose Consumption by Saccharomyces pastorianus Hybrids.
Saccharomyces pastorianus strains are hybrids of Saccharomyces cerevisiae and Saccharomyces eubayanus that have been domesticated for centuries in lager beer brewing environments. As sequences and structures of S. pastorianus genomes are being resolved, molecular mechanisms and evolutionary origins of several industrially relevant phenotypes remain unknown. This study investigates how maltotriose metabolism, a key feature in brewing, may have arisen in early S. eubayanus × S. cerevisiae hybrids. To address this question, we generated a nearly complete genome assembly of Himalayan S. eubayanus strains of the Holarctic subclade. This group of strains has been proposed to be the S. eubayanus subgenome origin of current S. pastorianus strains. The Himalayan S. eubayanus genomes harbored several copies of an S. eubayanusAGT1 (SeAGT1) α-oligoglucoside transporter gene with high sequence identity to genes encountered in S. pastorianus Although Himalayan S. eubayanus strains cannot grow on maltose and maltotriose, their maltose-hydrolase and SeMALT1 and SeAGT1 maltose transporter genes complemented the corresponding null mutants of S. cerevisiae Expression, in Himalayan S. eubayanus of a functional S. cerevisiae maltose metabolism regulator gene (MALx3) enabled growth on oligoglucosides. The hypothesis that the maltotriose-positive phenotype in S. pastorianus is a result of heterosis was experimentally tested by constructing an S. cerevisiae × S. eubayanus laboratory hybrid with a complement of maltose metabolism genes that resembles that of current S. pastorianus strains. The ability of this hybrid to consume maltotriose in brewer's wort demonstrated regulatory cross talk between subgenomes and thereby validated this hypothesis. These results support experimentally the new postulated hypothesis on the evolutionary origin of an essential phenotype of lager brewing strains and valuable knowledge for industrial exploitation of laboratory-made S. pastorianus-like hybrids.IMPORTANCES. pastorianus, an S. cerevisiae × S. eubayanus hybrid, is used for production of lager beer, the most produced alcoholic beverage worldwide. It emerged by spontaneous hybridization and colonized early lager brewing processes. Despite accumulation and analysis of genome sequencing data of S. pastorianus parental genomes, the genetic blueprint of industrially relevant phenotypes remains unresolved. Assimilation of maltotriose, an abundant sugar in wort, has been postulated to be inherited from the S. cerevisiae parent. Here, we demonstrate that although Asian S. eubayanus isolates harbor a functional maltotriose transporter SeAGT1 gene, they are unable to grow on α-oligoglucosides, but expression of S. cerevisiae regulator MAL13 (ScMAL13) was sufficient to restore growth on trisaccharides. We hypothesized that the S. pastorianus maltotriose phenotype results from regulatory interaction between S. cerevisiae maltose transcription activator and the promoter of SeAGT1 We experimentally confirmed the heterotic nature of the phenotype, and thus these results provide experimental evidence of the evolutionary origin of an essential phenotype of lager brewing strains.


Roles of Na,K-ATPase in early development and trophectoderm differentiation.
Before implantation into the uterine wall, the mammalian embryo undergoes a period of cell division, cell shape change, and cell differentiation leading to the formation of an outer epithelium, the trophectoderm. The trophectoderm is the part of the embryo that initiates uterine contact and, after transformation to become the trophoblast, uterine invasion. Similar to the kidney nephron, the trophectoderm is a transporting epithelium with distinct apical and basolateral membrane domains; its function is to facilitate transepithelial Na+ and fluid transport for blastocoel formation. That transport is driven by Na,K-adenosine triphosphatase (ATPase) localized in basolateral membranes of the trophectoderm. Preimplantation embryos express multiple alpha and beta subunit isoforms of Na,K-ATPase, potentially constituting multiple isozymes, but the basolaterally located alpha1beta1 isozyme appears to function uniquely to drive fluid transport. Embryos unable to express alpha1 subunits because of targeted deletion of the gene are able to form a blastocoel, but they fail to maintain their integrity and expire during the peri-implantation period. Preimplantation embryos also express the gamma subunit, a modulator of Na,K-ATPase activity, but targeted deletion of that gene did not reveal an essential developmental role. The preimplantation embryo offers a unique model for understanding the roles of Na,K-ATPase subunit isoforms in epithelial development and transepithelial transport.


EBNA-1 reactivity and HLA DRB1*1501 as statistically independent risk factors for multiple sclerosis: a case-control study.
The interaction between the two best documented risk factors (human leukocyte antigen [HLA] class II [DRB1*1501 positivity] and Epstein-Barr virus [elevated Epstein-Barr nuclear antigen 1 (EBNA-1) antibody reactivity]) for multiple sclerosis (MS) was studied in a case-control study of biobank samples from 109 MS cases and 212 matched referents. Multivariate logistic regression analysis showed that both were statistically significant in both sexes. HLA DRB1*1501-positive referents had higher EBNA-1 reactivity than HLA-negative referents. Less EBNA-1 reactivity was required to increase the MS risk in HLA DRB1*1501-positives than in HLA-negatives. We suggest that HLA DRB1*1501-positive individuals have an increased vulnerability to EBV-induced autoimmunity.


Induction of rapid and highly efficient expression of the human ND4 complex I subunit in the mouse visual system by self-complementary adeno-associated virus.
To demonstrate the high efficiency and rapidity of allotopic expression of a normal human ND4 subunit of complex I in the vertebrate retina using a self-complementary adeno-associated virus (scAAV) vector for ocular gene delivery to treat acute visual loss in Leber hereditary optic neuropathy (LHON). The nuclear-encoded human ND4 subunit fused to the P1 isoform of subunit C of adenosine triphosphate synthase (ATPc) mitochondrial targeting sequence and FLAG epitope was packaged in scAAV2 capsids or single-stranded (ss) AAV2 capsids. These constructs were injected into the vitreous cavities of mice. The contralateral eyes were injected with scAAV-green fluorescent protein (GFP). One week later, pattern electroretinograms and gene expression of the human ND4 subunit and GFP were evaluated. Quantitative analysis of ND4FLAG-injected eyes was assessed relative to Thy1.2-labeled retinal ganglion cells (RGCs). Pattern electroretinogram amplitudes remained normal in eyes inoculated with scAAV-ND4FLAG, ssAAV-ND4FLAG, and GFP. Confocal microscopy revealed the typical perinuclear mitochondrial expression of scAAV-ND4FLAG in almost the entire retinal flat mount. In contrast, scAAV-GFP expression was cytoplasmic and nuclear. Relative to Thy1.2-positive RGCs, quantification of scAAV-ND4FLAG-positive RGCs was 91% and that of ssAAV-ND4FLAG-positive RGCs was 51%. Treatment of acute visual loss due to LHON may be possible with a normal human ND4 subunit gene of complex I, mutated in most cases of LHON, when delivered by an scAAV vector. Clinical Relevance Unlike most retinal degenerations that result in slowly progressive loss of vision over many years, LHON due to mutated mitochondrial DNA results in apoplectic, bilateral severe and usually irreversible visual loss. For rescue of acute visual loss in LHON, a highly efficient and rapid gene expression system is required.


The groEL2 gene, but not groEL1, is required for biosynthesis of the secondary metabolite myxovirescin in Myxococcus xanthus DK1622.
Myxococcus xanthus DK1622 possesses two copies of the groEL gene: groEL1, which participates in development, and groEL2, which is involved in the predatory ability of cells. In this study, we determined that the groEL2 gene is required for the biosynthesis of the secondary metabolite myxovirescin (TA), which plays essential roles in predation. The groEL2-knockout mutant strain was defective in producing a zone of inhibition and displayed decreased killing ability against Escherichia coli, while the groEL1-knockout mutant strain exhibited little difference from the wild-type strain DK1622. HPLC revealed that deletion of the groEL2 gene blocked the production of TA, which was present in the groEL1-knockout mutant. The addition of exogenous TA rescued the inhibition and killing abilities of the groEL2-knockout mutant against E. coli. Analysis of GroEL domain-swapping mutants indicated that the C-terminal equatorial domain of GroEL2 was essential for TA production, while the N-terminal equatorial or apical domains of GroEL2 were not sufficient to rescue TA production of the groEL2 knockout.


Automatic detection of ST-T complex changes on the ECG using filtered RMS difference series: application to ambulatory ischemia monitoring.
A new detector is presented which finds changes in the repolarization phase (ST-T complex) of the cardiac cycle. It operates by applying a detection algorithm to the filtered root mean square (rms) series of differences between the beat segment (ST segment or ST-T complex) and an average pattern segment. The detector has been validated using the European ST-T database, which contains ST-T complex episodes manually annotated by cardiologists, resulting in sensitivity/positive predictivity of 85/86%, and 85/76%, for ST segment deviations and ST-T complex changes, respectively. The proposed detector has a performance similar to those which have a more complicated structure. The detector has the advantage of finding both ST segment deviations and entire ST-T complex changes thereby providing a wider characterization of the potential ischemic events. A post-processing stage, based on a cross-correlation analysis for the episodes in the rms series, is presented. With this stage subclinical events with repetitive pattern were found in around 20% of the recordings and improved the performance to 90/85%, and 89/76%, for ST segment and ST-T complex changes, respectively.


Kidney transplant rejection and tissue injury by gene profiling of biopsies and peripheral blood lymphocytes.
A major challenge for kidney transplantation is balancing the need for immunosuppression to prevent rejection, while minimizing drug-induced toxicities. We used DNA microarrays (HG-U95Av2 GeneChips, Affymetrix) to determine gene expression profiles for kidney biopsies and peripheral blood lymphocytes (PBLs) in transplant patients including normal donor kidneys, well-functioning transplants without rejection, kidneys undergoing acute rejection, and transplants with renal dysfunction without rejection. We developed a data analysis schema based on expression signal determination, class comparison and prediction, hierarchical clustering, statistical power analysis and real-time quantitative PCR validation. We identified distinct gene expression signatures for both biopsies and PBLs that correlated significantly with each of the different classes of transplant patients. This is the most complete report to date using commercial arrays to identify unique expression signatures in transplant biopsies distinguishing acute rejection, acute dysfunction without rejection and well-functioning transplants with no rejection history. We demonstrate for the first time the successful application of high density DNA chip analysis of PBL as a diagnostic tool for transplantation. The significance of these results, if validated in a multicenter prospective trial, would be the establishment of a metric based on gene expression signatures for monitoring the immune status and immunosuppression of transplanted patients.


Enhancement of drought tolerance in diverse Vicia faba cultivars by inoculation with plant growth-promoting rhizobacteria under newly reclaimed soil conditions.
Water deficit has devastating impacts on legume production, particularly with the current abrupt climate changes in arid environments. The application of plant growth-promoting rhizobacteria (PGPR) is an effective approach for producing natural nitrogen and attenuating the detrimental effects of drought stress. This study investigated the influence of inoculation with the PGPR Rhizobium leguminosarum biovar viciae (USDA 2435) and Pseudomonas putida (RA MTCC5279) solely or in combination on the physio-biochemical and agronomic traits of five diverse Vicia faba cultivars under well-watered (100% crop evapotranspiration [ETc]), moderate drought (75% ETc), and severe drought (50% ETc) conditions in newly reclaimed poor-fertility sandy soil. Drought stress substantially reduced the expression of photosynthetic pigments and water relation parameters. In contrast, antioxidant enzyme activities and osmoprotectants were considerably increased in plants under drought stress compared with those in well-watered plants. These adverse effects of drought stress reduced crop water productivity (CWP) and seed yield-related traits. However, the application of PGPR, particularly a consortium of both strains, improved these parameters and increased seed yield and CWP. The evaluated cultivars displayed varied tolerance to drought stress: Giza-843 and Giza-716 had the highest tolerance under well-watered and moderate drought conditions, whereas Giza-843 and Sakha-4 were more tolerant under severe drought conditions. Thus, co-inoculation of drought-tolerant cultivars with R. leguminosarum and P. putida enhanced their tolerance and increased their yield and CWP under water-deficit stress conditions. This study showed for the first time that the combined use of R. leguminosarum and P. putida is a promising and ecofriendly strategy for increasing drought tolerance in legume crops.


Divergent immune priming responses across flour beetle life stages and populations.
Growing evidence shows that low doses of pathogens may prime the immune response in many insects, conferring subsequent protection against infection in the same developmental stage (within-life stage priming), across life stages (ontogenic priming), or to offspring (transgenerational priming). Recent work also suggests that immune priming is a costly response. Thus, depending on host and pathogen ecology and evolutionary history, tradeoffs with other fitness components may constrain the evolution of priming. However, the relative impacts of priming at different life stages and across natural populations remain unknown. We quantified immune priming responses of 10 natural populations of the red flour beetle Tribolium castaneum, primed and infected with the natural insect pathogen Bacillus thuringiensis. We found that priming responses were highly variable both across life stages and populations, ranging from no detectable response to a 13-fold survival benefit. Comparing across stages, we found that ontogenic immune priming at the larval stage conferred maximum protection against infection. Finally, we found that various forms of priming showed sex-specific associations that may represent tradeoffs or shared mechanisms. These results indicate the importance of sex-, life stage-, and population-specific selective pressures that can cause substantial divergence in priming responses even within a species. Our work highlights the necessity of further work to understand the mechanistic basis of this variability.


Endoscopically-Assisted Scar-Free Midline Neck Mass Excision.
Traditional open surgery is indicated for the treatment of selected tumor subsites in the head and neck. However, it can cause major cosmetic problems and functional pathologies. The endoscopically-assisted transoral approach is increasingly preferred in some surgical fields due to its superior cosmetic and functional outcomes. Nonetheless, endoscopically-assisted transoral approach is not yet standard in the head and neck due to their anatomical complexity. The transoral surgical approach has been used for head and neck masses since the 1960s, and its application continues to evolve with changing disease conditions and recent innovations in surgical instruments. The potential for wide application of transoral surgery continues to be investigated, with a focus on minimizing occurrence of the complications. This review presents details of the surgical procedure and postoperative clinical outcomes, as well as endoscopically-assisted scar-free techniques for the resection of midline neck masses.


Clinical factors associated with sperm DNA fragmentation in male patients with infertility.
The clinical factors associated with sperm DNA fragmentation (SDF) were investigated in male patients with infertility. Fifty-four ejaculates from infertile Japanese males were used. Thirty-three and twenty-one were from the patients with varicoceles and idiopathic causes of infertility, respectively. We performed blood tests, including the serum sex hormone levels, and conventional and computer-assisted semen analyses. The sperm nuclear vacuolization (SNV) was evaluated using a high-magnification microscope. The SDF was evaluated using the sperm chromatin dispersion test (SCDt) to determine the SDF index (SDFI). The SDFI was compared with semen parameters and other clinical variables, including lifestyle factors. The SDFI was 41.3 ± 22.2% (mean ± standard deviation) and did not depend on the cause of infertility. Chronic alcohol use increased the SDFI to 49.6 ± 23.3% compared with 33.9 ± 18.0% in nondrinkers. The SDFI was related to adverse conventional semen parameters and sperm motion characteristics and correlated with the serum FSH level. The SNV showed a tendency to increase with the SDFI. The multivariate analysis revealed that the sperm progressive motility and chronic alcohol use were significant predictors of the SDF. The SCDt should be offered to chronic alcohol users and those with decreased sperm progressive motility.


Cellular reactions during drug-induced cleft palate.
Responses of the epithelial and mesenchymal cells of the developing hamster secondary palate to hadacidin, 5-fluorouracil, and hydrocortisone were analysed. Correlation of data on cellular response with morphological and biochemical observations suggested that several aspects of differentiation of palatal cells needs to be analysed before pathogenesis of teratogen-induced cleft palate can be defined.


Judgement of mood in music following right hemisphere damage.
Evidence on hemispheric specialization has implicated the right hemisphere as having a special role in the mediation of emotion. Since music is an area in which both cognitive and affective aspects of perception can be assessed, the aim of this study was to investigate the effect of right hemisphere damage on perception of emotional meaning or mood in music. An initial pilot study was conducted to select music on which normal subjects were consistent in their judgement of musical mood. The musical stimuli consisted of extracts of classical piano music. The technique used as a measure of musical mood was the Semantic Differential. Tracks of music and adjectival scales were selected for the experimental study in which 15 right hemisphere lesioned patients and normal controls were compared in their response to music. The right hemisphere group demonstrated a characteristic response in their judgement of mood in music. The results are discussed in terms of the role of pitch in judgement of mood in music and in terms of the relationship between music and language.


Estimation of gestational age of Nigerian newborn infants by measurement of thoracic spine length on chest radiographs.
Thoracic spine length was measured on chest radiographs of forty-nine Nigerian neonates of known gestational age. There is a strong correlation between gestational age and thoracic spine length (r = 0.93). The small-for-gestational-age (SGA) infants had thoracic spine lengths below two standard deviations (s.d.) of the mean for the appropriate-for-gestational-age infants. This method is useful for identifying SGA infants on chest radiographs particularly when they are too ill to be subjected to the usual physical and neurological assessment of gestational age.


Postprandial response of ghrelin and PYY and indices of low-grade chronic inflammation in lean young women with polycystic ovary syndrome.
The aim of the study were to answer the question 1.) Whether circulating pro-inflammatory markers of endothelial dysfunction and due to chronic low-grade inflammation of obesity, are altered in untreated lean, young relatively healthy polycystic ovary syndrome (PCOS) patients in comparison with healthy controls; 2.) Whether postprandial plasma concentration pattern of ghrelin and PYY can be predictable as risk factors for atherosclerosis and depend of obesity. Forty young women with PCOS were divided in two groups: 19 lean and 21 obese. The control group included 20 lean, healthy volunteers. Plasma total and active ghrelin, total PYY and PYY(3-36), serum adiponectin and insulin were measured using RIA technique, serum sCD40L, visfatin, sP-, sE-selectins, resistin by EIA. Composition of test meal was: 527 kcal total and consisted of 24.1% fat, 54.4% carbohydrate and 21.5% protein. Total and active ghrelin and total PYY were significantly lower in obese PCOS women, whereas active ghrelin was also significantly lower in lean PCOS women compared to controls. Postprandial plasma total ghrelin levels decrease were blunted in lean and obese compared to controls (12.8 % and 18.2% vs 28.2 %). Postprandial plasma active ghrelin decreased in lean and obese PCOS groups (49.9 % and 44.1 %) and controls (63.8 %). PCOS subjects exhibited smaller rises in postprandial levels of total PYY. Postprandial plasma PYY(3-36) levels increased in obese PCOS women (30.9 %) and controls (41%), whereas lean PCOS women exhibited blunted increase (11.5%). sCD40L levels increased, whereas adiponectin decreased in PCOS groups independently, whereas rise in visfatin, sE- and sP-selectin and the fall in adiponectin was associated with obesity. sP- and sE -selectins correlated positively with obesity. In summary, our study provides the first evidence that lean untreated young PCOS women contribute to the so called "pancreatic islet adaptation to insulin resistance" because of ghrelin and PYY profiles. We confirmed existing of low-grade chronic inflammation in early stage of visceral obesity in lean PCOS patients. The lost endogenous "islet adaptation to insulin resistance" may lead to endothelial dysfunction and promote acceleration of atherosclerosis.


Antimicrobial Peptide-Conjugated Hierarchical Antifouling Polymer Brushes for Functionalized Catheter Surfaces.
Catheter-related infection is a great challenge to modern medicine, which causes significant economic burden and increases patient morbidity. Hence, there is a great requirement for functionalized surfaces with inherently antibacterial properties and biocompatibility that prevent bacterial colonization and attachment of blood cells. Herein, we developed a strategy for constructing polymer brushes with hierarchical architecture on polyurethane (PU) via surface-initiated atom-transfer radical polymerization (SI-ATRP). Surface-functionalized PU (PU-DMH) was readily prepared, which comprised of poly(3-[dimethyl-[2-(2-methylprop-2-enoyloxy)ethyl]azaniumyl]propane-1-sulfonate) (PDMAPS) brushes as the lower layer and antimicrobial peptide-conjugated poly(methacrylic acid) (PMAA) brushes as the upper layer. The PU-DMH surface showed excellent bactericidal property against both Gram-positive and Gram-negative bacteria and could prevent accumulation of bacterial debris on surfaces. Simultaneously, the PU-DMH samples possessed good hemocompatibility and low cytotoxicity. Furthermore, the integrated antifouling and bactericidal properties of PU-DMH under hydrodynamic conditions were confirmed by an in vitro circulating model. The functionalized surface possessed persistent antifouling and bactericidal performances both under static and hydrodynamic conditions. The microbiological and histological results of animal experiments also verified the in vivo anti-infection performance. The present work might find promising clinical applications for preventing catheter-related infection.


Behavior and growth parameters of large white turkeys as affected by floor space and beak trimming. II. Females.
The effects of floor space allowance and beak trimming on behavior and growth performance of female Large White turkeys were studied. Floor space allowances were 5.6, 7.0, and 9.3 dm2/bird from 0 to 8 weeks of age and 14.0, 18.6, and 23.2 dm2/bird from 8 to 16 weeks of age. Floor space was adjusted by changing pen size to assure a constant number of birds per pen. Results showed that neither floor space allowance nor beak trimming had any significant effect of any of the variables under study through 8 weeks of age. At 16 weeks of age, growth, feed consumption, feed efficiency, feather scores, live market quality grades, and mortality were still unaffected. Beak trimming significantly reduced body weight gain, body weight, and feed consumption but had no significant effect on feed efficiency, feather scores, or mortality. Agonistic (aggressive) behavior was very low in this population of females. Of significance, however, were the nonaggressive behaviors of social "pecking" and "pecking and pulling" of feathers. Feather-pecking activity was unaffected by previous or subsequent floor space allowances through 16 weeks of age; however, the incidence of pecking and pulling of feathers was highest among females previously assigned the largest floor space allowance (9.3 dm2/bird) in the brooder house and subsequently allowed the smallest floor space allowance (14.0 dm2/bird) in the grower house. Beak trimming significantly increased the frequency of both "pecking" and "pecking and pulling" of feathers over that of their untrimmed contemporaries when the females were 12 and 16 weeks of age. Possible reasons for this are discussed.


On-road motor vehicle emissions and fuel consumption in urban driving conditions.
This paper reports on the analysis of on-road vehicle speed, emission, and fuel consumption data collected by four instrumented vehicles. Time-, distance-, and fuel-based average fuel consumption, as well as CO, HC, NOx, and soot emission factors, were derived. The influences of instantaneous vehicle speed on emissions and fuel consumption were studied. It was found that the fuel-based emission factors varied much less than the time- and distance-based emission factors as instantaneous speed changed. The trends are similar to the results obtained from laboratory tests. The low driving speed contributed to a significant portion of the total emissions over a trip. Furthermore, the on-road data were analyzed using the modal approach. The four standard driving modes are acceleration, cruising, deceleration, and idling. It was found that the transient driving modes (i.e., acceleration and deceleration) were more polluting than the steady-speed driving modes (i.e., cruising and idling) in terms of g/km and g/sec. These results indicated that the on-road emission measurement is feasible in deriving vehicle emissions and fuel consumption factors in urban driving conditions.


Effects of wogonin on the levels of cytokines and functions of leukocytes associated with NF-kappa B expression in Sprague-Dawley rats.
The objectives of the study were to investigate the effects of wogonin on the cytotoxicity, natural killer activity, macrophage phagocytosis, release of interleukin-1alpha (IL-1alpha) and tumor necrosis factor-alpha (TNF-alpha), the expression of nuclear factor-kappaB (NF-kappaB) in leukocytes from Sprague-Dawley rats. Wogonin was added to the medium and the levels of IL-1alpha and TNF-alpha in the medium were measured using enzyme-linked immunosorbent assay. Both IL-1alpha and TNF-alpha mRNAs were measured by reverse transcription polymerase chain reaction and the levels of NF-kappaB and I-kappaB were measured by Western blotting. Natural killer cell activity and macrophage phagocytosis were measured by flow cytometry. The results demonstrated that wogonin induced cytotoxicity in rat leukocytes and promoted the activities of natural killer cells and macrophages. Wogonin increased the IL-1alpha and TNF-alpha levels in the leukocytes in culture media dose-dependently. IL-1alpha and TNF-alpha mRNA elevations were also promoted by wogonin. The data suggests that wogonin may affect the IL-1alpha and TNF-alpha mRNA expressions by affecting the NF-kappaB in rat leukocytes.


Canadian cost-effectiveness analysis of solifenacin compared to oxybutynin immediate-release in patients with overactive bladder.
To estimate the cost effectiveness of solifenacin 5 mg/day compared to oxybutynin immediate-release (IR) 15 mg/day in patients with overactive bladder, from the perspective of the Canadian healthcare (payer) system. A Markov model was adapted to estimate the incremental cost per quality-adjusted life-year (QALY) of solifenacin and oxybutynin IR over a 1-year time horizon, based on efficacy and discontinuation data from the Canadian VECTOR (VEsicare in Comparison To Oxybutynin for oveRactive bladder patients) study. In the model, patients who discontinued treatment were offered tolterodine extended release 4 mg/day as second-line. Model robustness was tested using various sensitivity analyses. Utility values were derived from published literature; incontinence pads were included in a secondary analysis. In the base-case analysis, total costs over 1 year were CAN$695 and CAN$550 in the solifenacin and oxybutynin IR groups, respectively. When including incontinence pad costs, there was an incremental saving of CAN$1,831 per patient with solifenacin. Solifenacin was associated with an incremental QALY gain of 0.01 over 1 year. In the base-case analysis without incontinence pads, the incremental cost-utility ratio for solifenacin was CAN$14,092. Probabilistic analyses showed no overlap in the 95% confidence intervals for total costs or QALYs with or without incontinence pads. Solifenacin was cost effective in >90% of cases, based on a willingness-to-pay threshold of CAN$50,000 per additional QALY, irrespective of whether pad costs were included in the model. The most influential variables were the discontinuation rates and the cost of incontinence pads. Limitations of the analysis relate mainly to the fact that data in the VECTOR study were collected using a direct questioning approach, which might have increased the reporting of dry mouth. Solifenacin 5 mg/day was a cost-effective treatment compared with oxybutynin IR 15 mg/day. NCT00431041 (of the VECTOR study, upon which the analysis in this paper was based).


Microleakage at the interface between composite veneering material and precious alloy surface constructed either by new metallurgical sintering technique or the conventional casting technique.
The new metallurgical sintering technique in construction of precious metallic structure of the resin veneered restorations was introduced recently in the prosthetic dental work. This study compared the microleakage at the interface between photocured composite veneering material and gold platinum alloy surface constructed either by the new sintering technique or the conventional casting technique. Two chemically adhesive systems were used, a) silicoater and b) Lee metal primer, in order to retain the veneering material. The samples were subjected to thermal cycling and the procedures used in determination microleakage were the radioactive isotope and the autoradiograph methods. Two time intervals were compared 1--one month and 3--three months. It was concluded that the two different techniques used in construction of the metallic part showed no statistical significant difference in microleakage at their interface with the veneering material retained either by silicoater or Lee metal primer systems.


High-viability lyophilized Bacille Calmette-Guérin vaccine produced by deep-culture technique.
Evidence suggests that Bacille Calmette-Guérin (BCG) vaccine for use in cancer immunotherapy should have the following characteristics: high viability which is maintained on storage; high ratio of live to dead cells; high proportion of single cells; and low content of soluble antigen. The production of a vaccine with these characteristics was accomplished by use of a deep-culture technique. The medium was modified Proskauer and Beck medium containing Tween 80 and glucose. The mass culture was grown in a Wheaton double-side-arm bottle (6 liters of medium in an 8-liter container), aerated by means of an aquarium aerator and mixed by a magnetic stirrer. The culture was incubated 7 to 9 days at 37 degrees C, concentrated 11 to 15 times by ultrafiltration, diluted with equal parts of 25% lactose, and then lyophilized. The lyophilized ampoules, stored at -70 degrees C, were cultured at intervals ranging from 3 days to 450 days, and no loss in viability was observed. The mean number of viable BCG per ml of reconstituted vaccine was 8.75 log10. The viable count was 90% of the total bacterial count. Moreover, 85% of the cells were present as single bacilli.


Detrimental effect of postnatal blockade of N-methyl-D-aspartate receptors on sensorimotor gating is reversed by neuroleptic drugs.
Postnatal hypofunction of N-methyl-D-aspartate (NMDA) receptors leads to several behavioral deficits in adult rats resembling deficits typical of schizophrenia-like deficits of sensorimotor gating. Thus far, it is not known whether the above disruptions are sensitive to neuroleptic drugs. In order to verify the above model in pharmacological terms, we investigated whether deficits in the sensorimotor gating evoked by administration of NMDA receptor antagonists in the postnatal period is sensitive to neuroleptic drugs. We also investigated whether such treatment evoked alterations in the expression of dopamine D(1), D(2) and D(3) receptors in the nucleus accumbens, a key structure for dopamine-dependent alterations in sensorimotor gating. CGP 40116, a competitive antagonist of NMDA receptors was given in doses of 1.25 mg/kg on days 1, 3, 6 and 9; 2.5 mg/kg on days 12, 15 and 18; and 5 mg/kg on day 21 (all injections were sc). The efficacy of sensorimotor gating was tested on rats at the age of 60 days using a prepulse-induced inhibition of the startle reflex. In order to measure the expression of dopamine D(1), D(2) and D(3) receptors, we used quantitative autoradiography and tritiated ligands i.e. [(3)H]-SCH 23390, [(3)H]-Spiperone and [(3)H]-7-OH-DPAT, respectively. Haloperidol (0.1 mg/kg, sc), risperidone (1.0 mg/kg, sc) and clozapine (2.5 mg/kg, sc) reversed deficits of sensorimotor gating observed in adult rats evoked by the postnatal administration of CGP 40116. We also observed enhanced density of dopamine D(3), but not D(1) and D(2) receptors in the nucleus accumbens of CGP40116 treated rats. It is concluded that models of cognitive dysfunction, typical for schizophrenia based on postnatal administration of NMDA receptor antagonists, are sensitive to neuroleptic drugs and possibly not dependent on alteration in the density of dopaminergic receptors.


In vitro and in vivo toxicity evaluation of plant virus nanocarriers.
The use of biological self-assembling materials, plant virus nanoparticles in particular, appears very intriguing as it allows a great choice of symmetries and dimensions, easy chemical and biological engineering of both surface and/or internal cavity as well as safe and rapid production in plants. In this perspective, we present an initial evaluation of the safety profile of two structurally different plant viruses produced in Nicotiana benthamiana L. plants: the filamentous Potato virus X and the icosahedral Tomato bushy stunt virus. In vitro haemolysis assay was used to test the cytotoxic effects, which could arise by pVNPs interaction with cellular membranes, while early embryo assay was used to evaluate toxicity and teratogenicity in vivo. Data indicates that these structurally robust particles, still able to infect plants after incubation in serum up to 24h, have neither toxic nor teratogenic effects in vitro and in vivo. This work represents the first safety-focused characterization of pVNPs in view of their possible use as drug delivery carriers.


Randomized trial of high-dose chemotherapy and hematopoietic progenitor-cell support in operable breast cancer with extensive lymph node involvement: final analysis with 7 years of follow-up.
The aim of this study was to present an update of overall (OS) and disease-free survival (DFS) and to evaluate the correlation between outcome and pathological findings at surgery in a randomized trial of high-dose chemotherapy following neoadjuvant chemotherapy and surgery in high-risk breast cancer patients. Ninety-seven women <60 years of age with breast cancer and extensive axillary lymph node involvement received three courses of FE120C (5-fluorouracil 500 mg/m2, epirubicin 120 mg/m2, cyclophosphamide 500 mg/m2) followed by surgery. Eighty-one patients were randomized to receive either a fourth FE120C course alone or a fourth FE120C course followed by high-dose chemotherapy (cyclophosphamide 6 g/m2, thiotepa 480 mg/m2, carboplatin 1600 mg/m2). We performed a univariate analysis on possible prognostic factors and analyzed the sites of relapse. After a median follow-up of 6.9 years, 47 (48%) patients were alive, of whom 36 (38%) were without disease. Sixty patients relapsed after treatment. One patient died of myelodysplastic syndrome, without a relapse. In intention-to-treat analysis, the 5-year DFS rates were 47.5% in the conventional treatment arm and 49% in the high-dose arm, and the 5-year OS rates were 62.5% and 61%, respectively. In the univariate analysis, the clinical T-stage before chemotherapy and the number of tumor-positive axillary lymph nodes after induction chemotherapy (P = 0.027) were significant prognostic factors for OS. The same factors (both P = 0.06) plus the estrogen receptor (P = 0.08) were borderline significant factors for DFS. After a median follow-up of 6.9 years there was no difference in OS or DFS rates between the two treatment groups. The number of tumor-positive axillary lymph nodes after induction chemotherapy and the clinical T-stage before chemotherapy were significant factors for OS.


Speckle from branching vasculature: dependence on number density.
Purpose: Recent theories examine the role of the fractal branching vasculature as a primary site of Born scattering from soft normal tissues. These derivations postulate that the first-order statistics of speckle from soft tissue, such as the liver, thyroid, and prostate, will follow a Burr distribution with a power law parameter that can be related back to the underlying power law, which governs the branching network. However, the issue of scatterer spacing, or the number of cylindrical vessels per sample volume of the interrogating pulse, has not been directly addressed. Approach: Speckle statistics are examined with a 3D simulation that varies the number density and the governing power law parameter of an ensemble of different sized cylinders. Several in vivo liver scans are also analyzed for confirmation across different conditions. Results: The Burr distribution is found to be an appropriate model for the histogram of amplitudes from speckle regions, where the parameters track the underlying power law and scatterer density conditions. These results are also tested in a more general model of rat liver scans in normal versus abnormal conditions, and the resulting Burr parameters are also found to be appropriate and sensitive to underlying scatterer distributions. Conclusions: These preliminary results suggest that the classical Burr distribution may be useful in the quantification of scattering of ultrasound from soft vascularized tissues and as a tool in tissue characterization.


Uncovering and deciphering the pro-invasive role of HACE1 in melanoma cells.
HACE1 is an E3 ubiquitin ligase described as a tumour suppressor because HACE1-knockout mice develop multi-organ, late-onset cancers and because HACE1 expression is lost in several neoplasms, such as Wilms' tumours and colorectal cancer. However, a search of public databases indicated that HACE1 expression is maintained in melanomas. We demonstrated that HACE1 promoted melanoma cell migration and adhesion in vitro and was required for mouse lung colonisation by melanoma cells in vivo. Transcriptomic analysis of HACE1-depleted melanoma cells revealed an inhibition of ITGAV and ITGB1 as well changes in other genes involved in cell migration. We revealed that HACE1 promoted the K27 ubiquitination of fibronectin and regulated its secretion. Secreted fibronectin regulated ITGAV and ITGB1 expression, as well as melanoma cell adhesion and migration. Our findings disclose a novel molecular cascade involved in the regulation of fibronectin secretion, integrin expression and melanoma cell adhesion. By controlling this cascade, HACE1 displays pro-tumoural properties and is an important regulator of melanoma cell invasive properties.


Comparison of a Five-Year Survival and Cancer Recurrence between Laparoscopically Assisted and Open Colonic Resections due to Adenocarcinoma-a Single Centre Experience.
Background and objectives: When resecting colon adenocarcinoma, surgeons decide between the use of laparoscopically assisted and open surgery. Laparoscopic resection is known to have short-term benefits over an open operation. However, researchers are not as unified about the long-term findings. The aim of this research is to elaborate on five-year post-operative differences in survival and cancer recurrence between these two different approaches. Materials and methods: 74 enrolled patients were evaluated five years after a primary operation. We collected dates of deaths of deceased patients and time after operation of possible recurrences. Carcinoma staging was done by a pathologist after operation. Blood samples were taken before surgery in order to measure tumor markers (CA19-9 and CEA). Results: Survival after colonic adenocarcinoma surgery did not differ between the two different surgical approaches (p = 0.151). Recurrence of cancer was not associated with the type of operation (p = 0.532). Patients with recurrence had a 37.6 times greater hazard ratio of dying (95% CI: [12.0, 118]; p < 0.001). Advanced age adversely affected survival: patients aged <65 and 65 years had a 97%, and 57% survival rate, respectively. Patients with elevated tumor markers at operation had a 19.1 greater hazard ratio of dying (95% CI: [5.16, 70.4]; p<0.001). Patients with different TNM stages did not have any statistically significant differences in survival (HRII = 2.49; 95% CI: [0.67, 9.30]; pII = 0.173) (HRIII = 2.18; 95% CI: [0.58, 8.12]; pIII = 0.246) or recurrence (p = 0.097). Conclusion: The obtained results suggest that laparoscopic resection of colon cancer is not inferior from an oncologic point of view and results in a similar long-term survival and disease-free interval. Recurrence of carcinoma, older age at initial operation and elevated tumor markers, above a pre-set threshold at operation, were found to be independent factors of lower survival. We believe that the obtained results will be of benefit when choosing treatment for colon adenocarcinoma.


Firefly luciferin-activated rose bengal: in vitro photodynamic therapy by intracellular chemiluminescence in transgenic NIH 3T3 cells.
Photodynamic therapy (PDT) of cancer (1, 2) is a well-established treatment modality that uses light excitation of a photosensitive substance to produce oxygen-related cytotoxic intermediates, such as singlet oxygen or free radicals (3, 4). Although PDT is advantageous over other forms of cancer treatments because of its limited side effects, its main disadvantage is the poor accessibility of light to more deeply lying malignancies. External light sources such as lasers or lamps can be applied either noninvasively to reach tumors that lie well within the penetration depth of the light or in a minimally invasive fashion (interstitial treatments) in which optical fibers are placed intratumorally through needles. Even with the second approach, light distribution over the tumor is not homogeneous and nonidentified metastatic disease is left untreated. CL, the chemical production of light, is exemplified by firefly light emission mediated by the enzymatic (luciferase + ATP) oxidation of D-luciferin to oxyluciferin (5). This mobile light source is a targetable alternative to external sources of illumination. Here we show the in vitro photodynamic effect of rose bengal activated by intracellular generation of light, in luciferase-transfected NIH 3T3 murine fibroblasts.


Purification of a major membrane protein of Toxoplasma gondii by immunoabsorption with a monoclonal antibody.
The principal iodinatable surface protein (P30) of our cloned RH strain of Toxoplasma gondii has an apparent molecular weight of 30,000, as measured by acrylamide gel electrophoresis in the presence of sodium dodecyl sulfate under reducing conditions. Monoclonal antibody B specifically immunoprecipitated protein P30 from a detergent extract of surface radioiodinated T. gondii. Monoclonal antibody B in the presence of complement was also parasiticidal for T. gondii, and this parasiticidal effect could be blocked by protein P30. Monoclonal antibody B was purified from mouse ascitic fluid and linked to cyanogen bromide-activated Sepharose. The resulting immunoabsorbent was used to purify 1.7 mg of protein P30 from a large number of parasites. The efficiency of recovery of protein P30 was measured by assays of radioactivity and of parasiticidal blocking activity. Protein P30 represented 3 to 5% of the total protein. It is also present in a recently isolated strain of T. gondii. A convalescent human antitoxoplasma serum immunoprecipitated radiolabeled protein P30. Three convalescent antisera when quantitated by an ELISA test had a high anti-protein P30 titer. Charge shift electrophoresis showed that protein P30 has an extensive hydrophobic region and thus is probably an integral membrane protein. Electrophoresis under nonreducing conditions showed no evidence that protein P30 exists as a disulfide linked homo- or heterodimer, although it probably has intramolecular disulfide bonds.


Collaborative and Overinvolved Parenting Differentially Predict Outcomes in Adolescents With Type 1 Diabetes.
The current study examined the impact of collaborative and intrusive parenting on depression and glycemic control in children with Type 1 diabetes (T1D). Research examining the association between parenting and child adjustment in this population has been limited by a reliance primarily on questionnaire data and cross-sectional analyses. To address these gaps, the current study used an observational coding system to measure the effects of parenting on child adjustment over a 1-year period. Youth (10 to 16 years old) with T1D and their mothers (N = 81) were recruited from an outpatient pediatrics diabetes clinic. Mothers' symptoms of anxiety and depression and children's depressive symptoms were assessed by self- reports; parenting behaviors were assessed via video-recorded observations coded using the Iowa Family Interaction Rating Scales; and adolescents' glycosylated hemoglobin (HbA1c) was obtained from medical records. Bivariate correlations and linear regression analyses revealed that higher levels of observed collaborative parenting were related to significantly lower HbA1c 12 months later, and higher levels of observed overinvolved parenting were related to significantly greater child depressive symptoms 12 months later. Further, age and treatment type moderated the relation between overinvolved parenting and child depressive symptoms. Collaborative and overinvolved parenting appears important for adolescents in predicting both psychological and health-related outcomes over time. Parenting behaviors may serve as an important target for future interventions to enhance adjustment in these children. (PsycINFO Database Record


Temporal compositional shifts in an activated sludge microbiome during estrone biodegradation.
Microbial biodegradation is a key process for the removal of estrogens during wastewater treatment. At least four degradation pathways for natural estrogens have been proposed. However, major estrogen degraders and the occurrence of different estrogen biodegradation pathways in wastewater treatment plants have been rarely investigated. This study was conducted to elucidate estrone biodegradation pathway and to identify key estrone-degrading bacteria in activated sludge from a major wastewater treatment plant in Bahrain. The biodegradation experiments were performed in activated sludge microcosms supplemented with estrone. Sludge samples were retrieved at time intervals to analyze the biodegradation metabolites and the temporal shifts in the bacterial community composition. Chemical analysis revealed the biodegradation of more than 90% of the added estrone within 6 days, and the compounds 4-hydroxyestrone and pyridinestrone acid, which are typical markers of the 4,5-seco pathway of aerobic estrone biodegradation, were detected. Temporal shifts in the relative abundance of bacteria were most prominent among members of Proteobacteria and Bacteroidetes. While the alphaproteobacterial genera Novosphingobium and Sphingoaurantiacus were significantly enriched (from ≤ 6% to an average of 31%) in the estrone-amended activated sludge after 2 days of incubation, the bacteroidete Pedobacter was uniquely detected in these microcosms at day 10. The relative abundance of Polyangia (Nannocyctis) increased to an average of 10 ± 0.4% in the estrone-amended activated sludge after 4 days of incubation. Enrichment cultivation of bacteria from the activated sludge on estrone resulted in a mixed culture that was capable of degrading estrone. An estrone-degrading strain was isolated from this mixed culture and was affiliated with the known estrogen-degrading Alphaproteobacteria Sphingobium estrogenivorans. We conclude that estrone degradation in the activated sludge from the studied wastewater treatment plant proceeds via the 4,5-seco pathway and is most likely mediated by alphaproteobacterial taxa.


Comparison of Parametric Survival Extrapolation Approaches Incorporating General Population Mortality for Adequate Health Technology Assessment of New Oncology Drugs.
Survival extrapolation of trial outcomes is required for health economic evaluation. Generally, all-cause mortality (ACM) is modeled using standard parametric distributions, often without distinguishing disease-specific/excess mortality and general population background mortality (GPM). Recent National Institute for Health and Care Excellence guidance (Technical Support Document 21) recommends adding GPM hazards to disease-specific/excess mortality hazards in the log-likelihood function ("internal additive hazards"). This article compares alternative extrapolation approaches with and without GPM adjustment. Survival extrapolations using the internal additive hazards approach (1) are compared to no GPM adjustment (2), applying GPM hazards once ACM hazards drop below GPM hazards (3), adding GPM hazards to ACM hazards (4), and proportional hazards for ACM versus GPM hazards (5). The fit, face validity, mean predicted life-years, and corresponding uncertainty measures are assessed for the active versus control arms of immature and mature (30- and 75-month follow-up) multiple myeloma data and mature (64-month follow-up) breast cancer data. The 5 approaches yielded considerably different outcomes. Incremental mean predicted life-years vary most in the immature multiple myeloma data set. The lognormal distribution (best statistical fit for approaches 1-4) produces survival increments of 3.5 (95% credible interval: 1.4-5.3), 8.5 (3.1-13.0), 3.5 (1.3-5.4), 2.9 (1.1-4.5), and 1.6 (0.4-2.8) years for approaches 1 to 5, respectively. Approach 1 had the highest face validity for all data sets. Uncertainty over parametric distributions was comparable for GPM-adjusted approaches 1, 3, and 4, and much larger for approach 2. This study highlights the importance of GPM adjustment, and particularly of incorporating GPM hazards in the log-likelihood function of standard parametric distributions.


Food Losses and Waste: A Needed Assessment for Future Policies.
About one third of food produced for human consumption is lost or wasted. For this reason, food losses and waste has become a key priority within worldwide policy circles. This is a major global issue that not only threatens the viability of a sustainable food system but also generates negative externalities in environmental terms. The avoidance of this forbidding wastage would have a positive economic impact on national economies in terms of resource savings. In this paper we look beyond this somewhat traditional resource savings angle and we shift the focus to explore the distributional consequences of food losses and waste reduction using a resource constrained modeling perspective. The impact due to the behavioral shift of each household is therefore explained by two factors. One is the amount of resources saved when the behavioral shift takes place, whereas the other one has to do with the position of households in the food supply chain. By considering the whole supply chain, instead of the common approach based only in reducing waste by consumers, we enrich the empirical knowledge of this issue and improve the quantification of its economic impact. We examine data for three EU countries that present different economic structures (Germany, Spain and Poland) so as to have a broader and more robust viewpoint of the potential results. We find that distributional effects are different for consumers and producers and also across countries. Our results could be useful for policymakers since they indicate that policies should not be driven merely by the size waste but rather on its position within the food supply chain.


Long circulating half-life and high tumor selectivity of the photosensitizer meta-tetrahydroxyphenylchlorin conjugated to polyethylene glycol in nude mice grafted with a human colon carcinoma.
In a mode of nude mice bearing a human colon carcinoma xenograft, the biodistribution and tumor localization of metatetrahydroxyphenylchlorin (m-THPC) coupled to polyethylene glycol (PEG) were compared with those of the free form of this photosensitizer used in photodynamic therapy (PDT). At different times after i.v. injection of both forms of 125I-labeled photosensitizer, m-THPC-PEG gave on average a 2-fold higher tumor uptake than free m-THPC. In addition, at early times after injection, m-THPC-PEG showed a 2-fold longer blood circulating half-life and a 4-fold lower liver uptake than free m-THPC. The tumor to normal tissue ratios of radioactivity concentrations were always higher for m-THPC-PEG than for free m-THPC at any time point studied from 2 to 96 hr post-injection. Significant coefficients of correlation between direct fluorescence measurements and radioactivity counting were obtained within each organ tested. Fluorescence microscopy studies showed that m-THPC-PEG was preferentially localized near the tumor vessels, whereas m-THPC was more diffusely distributed inside the tumor tissue. To verify whether m-THPC-PEG conjugate remained phototoxic in vivo, PDT experiments were performed 72 hr after injection and showed that m-THPC-PEG was as potent as free m-THPC in the induction of tumor regression provided that the irradiation does for m-THPC-PEG conjugate was adapted to a well-tolerated 2-fold higher level. The overall results demonstrate first the possibility of improving the in vivo tumor localization of a hydrophobic dye used for PDT by coupling it to PEG and second that a photosensitizer conjugated to a macromolecule can remain phototoxic in vivo.


The health care costs of violence against women.
The aim of this study is to analyze the health care costs of violence against women. For the study, we used a register-based approach where we identified victims of violence and assessed their actual health care costs at individual level in a bottom-up analysis. Furthermore, we identified a reference population. We computed the attributable costs, that is, the excess health care costs for victims compared to an identified reference population of nonvictims. Only costs within the health care sector were included, that is, somatic and psychiatric hospital costs, costs within the primary health care sector and costs of prescription pharmaceuticals. We estimated the attributable health care costs of violence against women in Denmark, using a generalized linear model where health care costs were modeled as a function of age, childbirth, and exposure to violence. In addition we tested whether socioeconomic status, multiple episodes of violence, and psychiatric contacts had any impact on health care costs. We found that the health care costs were about €1,800 higher for victims of violence than for nonvictims per year, driven mostly by higher psychiatric costs and multiple episodes of violence.


Management strategy influences coral oxidative stress responses in a marine protected area in the Southwestern Atlantic.
Coral reefs are experiencing accelerated degradation due to global and local stressors. The understanding of how corals cope with these disturbances is urgent. We focused on elucidating antioxidant capacity responses of the Mussismilia harttii and Siderastrea sp. corals, in reefs with use management in a marine protected area. We tested whether the activity of antioxidant enzymes in healthy colonies is higher at multiple-use reefs than at no-take reef, and whether the activity of antioxidant enzymes is higher for bleached than for healthy Siderastrea sp. colonies. Lipid peroxidation and enzymatic activity found in bleached colonies evidence chronic stress and cellular damage not related to thermal anomalies. Chronic stress in healthy colonies was also found but responses differed among species, being higher at multiple use reefs, mainly for Siderastrea sp. We highlight the role of the local conservation actions in the integrity of coral physiology and reef resilience under global climate changes.


Myocardial protection from ischemic arrest: potassium and verapamil cardioplegia.
Prolonged normothermic myocardial ischemic arrest results in myocardial dysfunction. This study has investigated the technique of preserving myocardial function by a single bolus intracoronary infusion of combination potassium and verapamil at the onset of ischemic arrest. Sixty-one dogs underwent cardiopulmonary bypass with 60 min of ischemic arrest: 25 received no myocardial protection, 12 received a single intracoronary bolus of KCl, 12 received combination verapamil and KCl, and 12 received verapamil alone. Following the ischemic arrest, hearts protected by combination of potassium and verapamil demonstrated better survival evidenced by the ability of all 12 dogs to resume normal hemodynamic function. The hemodynamic function in the combination potassium and verapamil group also demonstrated better cardiac output, left ventricular dF/dt, and myocardial segment shortening than survivors in the other groups. Subsarcolemmal (SSL) and intermyofibrillar IMF) mitochondria were isolated from these hearts and function evaluated. NADH-linked oxygen consumption was impaired as was calcium transport in the SSL from unprotected ischemic hearts. Intermyofibrillar mitochondria were not different from control or sham. The hearts protected by verapamil and potassium demonstrated normal mitochondrial function.


A comparison of the isometric force fatigue-recovery profile in two posterior chain lower limb tests following simulated soccer competition.
To evaluate the reliability of isometric peak force (IPF) in a novel "long-length" 90°Hip:20°Knee (90:20) strength test and to compare the simulated soccer match induced fatigue-recovery profile of IPF in this test with that of an isometric 90°Hip:90°Knee (90:90) position test. Twenty semi-professional soccer players volunteered for the study of which 14 participated in the first part of the study which assessed 90:20 reliability (age = 21.3 ± 2.5 years, height = 1.79 ± 0.07 m, body mass = 73.2 ± 8.8 kg), while 17 completed the second part of the study evaluating fatigue-recovery (age 21.2±2.4 yrs., height = 180 ± 0.09 m, body mass 73.8 ± 8.9 kg). We evaluated the inter-session reliability of IPF in two 90:20 test protocols (hands on the wall (HW); and hands on chest (HC)) both performed on two occasions, 7 days apart. We then assessed 90:20 (HC) and 90:90 IPF immediately before (PRE) and after (POST) after a simulated soccer match protocol (BEAST90mod) and 48 (+48 h) and 72 hours (+72 h) later. Part one: the 90:20 showed moderate to high overall reliability (CV's of 7.3% to 11.0%) across test positions and limbs. CV's were lower in the HW than HC in the dominant (7.3% vs 11.0%) but the opposite happened in the non-dominant limb where CV's were higher in the HW than HC (9.7% vs 7.3%). Based on these results, the HC position was used in part two of the study. Part two: 90:20 and 90:90 IPF was significantly lower POST compared to PRE BEAST90mod across all testing positions (p<0.001). IPF was significantly lower at +48 h compared to PRE in the 90:20 in both limbs (Dominant: p<0.01,Non-dominant: p≤0.05), but not in the 90:90. At +72 h, IPF was not significantly different from PRE in either test. Simple to implement posterior IPF tests can help to define recovery from competition and training load in football and, potentially, in other multiple sprint athletes. Testing posterior chain IPF in a more knee extended 90:20 position may provide greater sensitivity to fatigue at 48 h post simulated competition than testing in the 90:90 position, but also may require greater degree of familiarization due to more functional testing position.


Homoeopathic therapy in rheumatoid arthritis: evaluation by double-blind clinical therapeutic trial.
Twenty-three patients with rheumatoid arthritis on orthodox first-line anti-inflammatory treatment plus homeopathy were compared wtih a similar group of twenty-three patients on orthodox first-line treatment plus an inert preparation. There was a significant improvement in subjective pain, articular index, stiffness and grip strength in those patients receiving homoeopathic remedies whereas there was no significant change in the patients who received placebo. Two physicians were involved in prescribing for the patients and there were no significant differences in the results which they obtained. No side effects were observed with the homoeopathic remedies.


Comparisons of phase I and phase II in vitro hepatic enzyme activities of human, dog, rhesus monkey, and cynomolgus monkey.
The metabolism of probe substrates of phase I and phase II enzymes in vitro were compared in hepatic subcellular fractions from humans, cynomolgus monkeys, rhesus monkeys, and beagle dogs. These studies were undertaken to compare the suitability of these species as models of metabolism in drug development. Eight cytochrome P450-dependent activities were measured in microsomal incubations: ethoxyresorufin O-deethylase, coumarin 7-hydroxylase, tolbutamide 4-hydroxylase, S-mephenytoin 4'-hydroxylase, bufuralol 1'-hydroxylase, N-nitrosodimethylamine N-demethylase, midazolam 1'-hydroxylase, and erythromycin N-demethylase. Seven phase II activities were determined in the appropriate subcellular fractions:acetaminophen UDP-glucurono-syltransferase, acetaminophen sulfotransferase, 17 alpha-ethinylestradiol UDP-glucuronosyltransferase, 17 alpha-ethinylestradiol sulfotransferase, 6-mercaptopurine methylase, dichloronitrobenzene (DCNB) glutathione S-transferase, and isoniazid N-acetylase. Hepatic subcellular fractions from cynomolgus and rhesus monkeys showed significantly higher activities than those from humans for ethoxyresorufin O-deethylase, bufuralol 1'-hydroxylase, midazolam 1'-hydroxylase, erythromycin N-demethylase, acetaminophen UDP-glucuronosyltransferase, acetaminophen sulfotransferase, and tolbutamide 4-hydroxylase. Cynomolgus monkey had higher activity than humans and rhesus monkeys for S-mephenytoin 4'-hydroxylase erythromycin N-demethylase. Rhesus monkey and human cytosol displayed an apparent genetic polymorphism in the N-acetylation of isoniazid, whereas cynomolgus monkey cytosol did not. All other monkey activities were not significantly different than human. Dog subcellular fractions showed higher activity than humans for midazolam 1'-hydroxylase, erythromycin N-demethylase, acetaminophen UDP-glucuronosyltransferase, acetaminophen sulfotransferase, 17 alpha-ethinylestradiol sulfotransferase, and DCNB glutathione S-transferase. Furthermore, dog samples had significantly lower activity for coumarin 7-hydroxylase and 6-mercaptopurine methylase, and no detectable activity for tolbutamide 4-hydroxylase or isoniazid N-acetylase. All other activities were not significantly different from human. These results reveal minor differences between the cynomolgus and rhesus monkey in drug metabolism capacities in vitro, but both species are generally more metabolically active than humans in both phase I and phase II metabolism, whereas dogs had more diverse deviations from humans.


Differences in the folding transition state of ubiquitin indicated by phi and psi analyses.
We compare the folding transition state (TS) of ubiquitin previously identified by using psi analysis to that determined by using analysis. Both methods attempt to identify interactions and their relative populations at the rate-limiting step for folding. The TS ensemble derived from psi analysis has an extensive native-like chain topology, with a four-stranded beta-sheet network and a portion of the major helix. According to analysis, however, the TS is much smaller and more polarized, with only a local helix/hairpin motif. We find that structured regions can have values far from unity, the canonical value for such sites, because of structural relaxation of the TS. Consequently, these sites may be incorrectly interpreted as contributing little to the structure of the TS. These results stress the need for caution when interpreting and drawing conclusions from analysis alone and highlight the need for more specific tools for examining the structure and energetics of the TS ensemble.


The climate and health benefits from intensive building energy efficiency improvements.
Intensive building energy efficiency improvements can reduce emissions from energy use, improving outdoor air quality and human health, but may also affect ventilation and indoor air quality. This study examines the effects of highly ambitious, yet feasible, building energy efficiency upgrades in the United States. Our energy efficiency scenarios, derived from the literature, lead to a 6 to 11% reduction in carbon dioxide emissions and 18 to 25% reductions in particulate matter (PM2.5) emissions in 2050. These reductions are complementary with a carbon pricing policy on electricity. However, our results also point to the importance of mitigating indoor PM2.5 emissions, improving PM2.5 filtration, and evaluating ventilation-related policies. Even with no further ventilation improvements, we estimate that intensive energy efficiency scenarios could prevent 1800 to 3600 premature deaths per year across the United States in 2050. With further investments in indoor air quality, this can rise to 2900 to 5100.


Rabbit anti-rat thymocyte immunoglobulin preserves renal function during ischemia/reperfusion injury in rat kidney transplantation.
Ischemia/reperfusion (I/R) injury is an important cause of renal graft dysfunction in humans. Increases in cold and warm ischemia times lead to a higher risk of early post-transplant complications including delayed graft function and acute rejection. Moreover, prolonged cold ischemia is a predictor of long-term kidney graft loss. The protective effect of rabbit anti-rat thymocyte immunoglobulin (rATG) was evaluated in a rat model of I/R injury following syngeneic kidney transplantation. Serum creatinine concentration was evaluated at 16 h and 24 h post-transplant. Animals were sacrificed 24 h post-transplant for evaluation of histology, infiltrating leukocytes, nitrotyrosine staining, and apoptosis. rATG was effective in preventing renal function impairment, tissue damage and tubular apoptosis associated with I/R only when was given 2 h before transplantation but not at the time of reperfusion. Pretransplant rATG treatment of recipient animals effectively reduced the amount of macrophages, CD4(+), CD8(+) T cells and LFA-1(+) cells infiltrating renal graft subjected to cold ischemia as well as granzyme-B expression within ischemic kidney. On the other hand, granulocyte infiltration and oxidative stress were not modified by rATG. If these results will be translated into the clinical setting, pretransplant administration of Thymoglobuline(®) could offer the additional advantage over peri-transplant administration of limiting I/R-mediated kidney graft damage.


Chiral disubstituted piperidinyl ureas: a class of dual diacylglycerol lipase-α and ABHD6 inhibitors.
Inhibitors of diacylglycerol lipases and α,β-hydrolase domain containing protein 6 (ABHD6) are potential leads for the development of therapeutic agents for metabolic and neurodegenerative disorders. Here, we report the enantioselective synthesis and structure activity relationships of triazole ureas featuring chiral, hydroxylated 2-benzylpiperidines as dual inhibitors of DAGLα and ABHD6. The chirality of the carbon bearing the C2 substituent, as well as the position of the hydroxyl (tolerated at C5, but not at C3) has profound influence on the inhibitory activity of both DAGLα and ABHD6, as established using biochemical assays and competitive activity-based protein profiling on mouse brain extracts.


Incidence of device-related infection in 97 750 patients: clinical data from the complete Danish device-cohort (1982-2018).
Device-related infection (DRI) is a severe complication to cardiac implantable electronic devices (CIED) therapy. Device-related infection incidence and its risk factors differ between previous studies. We aimed to define the long-term incidence and incidence rates of DRI for different types of CIEDs in the complete Danish device-cohort and identify patient-, operation- and device-related risk factors for DRI. From the Danish Pacemaker (PM) and implantable cardioverter-defibrillator (ICD) Register, we included consecutive Danish patients undergoing CIED implantation or reoperation from January 1982 to April 2018, resulting in 97 750 patients, 128 045 operations and follow-up of in total 566 275 device years (DY). We identified 1827 DRI causing device removals. Device-related infection incidence during device lifetime was 1.19% (1.12-1.26) for PM, 1.91% (1.71-2.13) for ICD, 2.18% (1.78-2.64) for cardiac resynchronization therapy (CRT)-pacemakers (CRT-P), and 3.35% (2.92-3.83) for CRT-defibrillators (CRT-D). Incidence rates in de novo implantations were 2.04/1000 DY for PM, 3.84 for ICD, 4.38 for CRT-P, and 6.76 for CRT-D. Using multiple-record and multiple-event per subject proportional hazard analysis, we identified implantation of complex devices (ICD and CRT), reoperations, prior DRI, male sex, and younger age as significantly associated with higher DRI risk. Overall risk of infection was low in PM implantations but considerably higher in CRT systems and after reinterventions. These data support the importance of evaluating all patients considered for CIED therapy thoroughly, in order to identify potential modifiable risk factors and reduce the risk of early reoperations.


[Risk factors for ischemic stroke. I. Conventional risk factors].
Stroke, together with ischemic cardiopathy, may be considered to be a vascular epidemic in the developed world, where it is a major social and health problem. In Spain it is the commonest cause of death in women and the second commonest in men. In this article we review the risk factors which cannot be modified (age, sex, inheritance, race, geography and climate) and the well-established risk factors which can be modified (arterial hypertension, cardiopathies with a high risk of embolization, markers for atherosclerosis, diabetes mellitus and transient ischemic accidents). A wide understanding of these risk factors permits the identification of persons with a high risk of having an ischemic stroke. Therefore preventive and therapeutic measures to reduce individual risk from strokes (and their recurrence) and hence reduce morbimortality from this, can be designed.


Effect of catchment characteristics on aquatic carbon export from a boreal catchment and its importance in regional carbon cycling.
Inland waters transport and emit into the atmosphere large amounts of carbon (C), which originates from terrestrial ecosystems. The effect of land cover and land-use practises on C export from terrestrial ecosystems to inland waters is not fully understood, especially in heterogeneous landscapes under human influence. We sampled for dissolved C species in five tributaries with well-determined subcatchments (total size 174.5 km(2)), as well as in various points of two of the subcatchments draining to a boreal lake in southern Finland over a full year. Our aim was to find out how land cover and land-use affect C export from the catchments, as well as CH4 and CO2 concentrations of the streams, and if the origin of C in stream water can be determined from proxies for quality of dissolved organic matter (DOM). We further estimated the gas evasion from stream surfaces and the role of aquatic fluxes in regional C cycling. The export rate of C from the terrestrial system through an aquatic conduit was 19.3 g C m(-2) (catchment) yr(-1), which corresponds to 19% of the estimated terrestrial net ecosystem exchange of the catchment. Most of the C load to the recipient lake consisted of dissolved organic carbon (DOC, 6.1 ± 1.0 g C m(-2) yr(-1)); the share of dissolved inorganic carbon (DIC) was much smaller (1.0 ± 0.2 g C m(-2) yr(-1)). CO2 and CH4 emissions from stream and ditch surfaces were 7.0 ± 2.4 g C m(-2) yr(-1) and 0.1 ± 0.04 g C m(-2) yr(-1), respectively, C emissions being thus equal with C load to the lake. The proportion of peatland in the catchment and the drainage density of peatland increased DOC in streams, whereas the proportion of agricultural land in the catchment decreased it. The opposite was true for DIC. Drained peatlands were an important CH4 source for streams.


Language, speech sound production, and cognition in three-year-old children in relation to otitis media in their first three years of life.
As part of a prospective study of possible effects of early-life otitis media on speech, language, cognitive, and psychosocial development, we tested relationships between children's cumulative duration of middle ear effusion (MEE) in their first 3 years of life and their scores on measures of language, speech sound production, and cognition at 3 years of age. We enrolled 6350 healthy infants by 2 months of age who presented for primary care at 1 of 2 urban hospitals or 1 of 2 small town/rural and 4 suburban private pediatric practices. We intensively monitored the children's middle ear status by pneumatic otoscopy, supplemented by tympanometry, throughout their first 3 years of life; we monitored the validity of the otoscopic observations on an ongoing basis; and we treated children for otitis media according to specified guidelines. Children who met specified minimum criteria regarding the persistence of MEE became eligible for a clinical trial in which they were assigned randomly to undergo tympanostomy tube placement either promptly or after a defined extended period if MEE remained present. From among those remaining, we selected randomly, within sociodemographic strata, a sample of 241 children who represented a spectrum of MEE experience from having no MEE to having MEE whose cumulative duration fell just short of meeting randomization criteria. In subjects so selected, the estimated duration of MEE ranged from none to 65.6% of the first year of life and 44.8% of the first 3 years of life. In these 241 children we assessed language development, speech sound production, and cognition at 3 years of age, using both formal tests and conversational samples. We found weak to moderate, statistically significant negative correlations between children's cumulative durations of MEE in their first year of life or in age periods that included their first year of life, and their scores on formal tests of receptive vocabulary and verbal aspects of cognition at 3 years of age. However, the percent of variance in these scores explained by time with MEE in the first year of life beyond that explained by sociodemographic variables ranged only from 1.2% to 2.9%, and the negative correlations were concentrated in the subgroup of children whose families had private health insurance (rather than Medicaid). We found no significant correlations in the study population as a whole or in any subgroup between time with MEE during antecedent periods and children's scores on measures of spontaneous expressive language, speech sound production, or other measured aspects of cognition. In contrast, by wide margins, scores on all measures were consistently highest among the most socioeconomically advantaged children and lowest among the most socioeconomically disadvantaged children. Our findings suggest either that persistent early-life MEE actually causes later small, circumscribed impairments of receptive language and verbal aspects of cognition in certain groups of children or that unidentified, confounding factors predispose children both to early-life otitis media and to certain types of developmental impairment. Findings in the randomized clinical trial component of the larger study should help distinguish between causality and confounding as explanations for our findings.language, speech, cognition, development, otitis media, middle ear effusion.


Aquaporin-1 promotes angiogenesis, fibrosis, and portal hypertension through mechanisms dependent on osmotically sensitive microRNAs.
Changes in hepatic vasculature accompany fibrogenesis, and targeting angiogenic molecules often attenuates fibrosis in animals. Aquaporin-1 (AQP1) is a water channel, overexpressed in cirrhosis, that promotes angiogenesis by enhancing endothelial invasion. The effect of AQP1 on fibrogenesis in vivo and the mechanisms driving AQP1 expression during cirrhosis remain unclear. The purpose of this study was to test the effect of AQP1 deletion in cirrhosis and explore mechanisms regulating AQP1. After bile duct ligation, wild-type mice overexpress AQP1 that colocalizes with vascular markers and sites of robust angiogenesis. AQP1 knockout mice demonstrated reduced angiogenesis compared with wild-type mice, as evidenced by immunostaining and endothelial invasion/proliferation in vitro. Fibrosis and portal hypertension were attenuated based on immunostaining, portal pressure, and spleen/body weight ratio. AQP1 protein, but not mRNA, was induced by hyperosmolality in vitro, suggesting post-transcriptional regulation. Endothelial cells from normal or cirrhotic mice were screened for microRNA (miR) expression using an array and a quantitative PCR. miR-666 and miR-708 targeted AQP1 mRNA and were decreased in cirrhosis and in cells exposed to hyperosmolality, suggesting that these miRs mediate osmolar changes via AQP1. Binding of the miRs to the untranslated region of AQP1 was assessed using luciferase assays. In conclusion, AQP1 promotes angiogenesis, fibrosis, and portal hypertension after bile duct ligation and is regulated by osmotically sensitive miRs.


Imaging of Endothelial Cell Dynamic Behavior in Zebrafish.
In recent years, use of the zebrafish embryo as a model organism to study vascular development in vivo has provided valuable insights into the genetic and cellular events shaping the embryonic vasculature. In this chapter, we aim to present the methods for the measurement of some of the most commonly investigated dynamic parameters in endothelial cells during developmental angiogenesis, namely, migration speed and acceleration, filopodia extension, front-rear polarity, cell cycle progression, membrane deformations, and junctional rearrangements. We also offer suggestions on how to deal with the most common imaging and quantifications challenges faced when acquiring and quantifying endothelial cell dynamic behavior in vivo.We intend this section to serve as an experience-based imaging primer for scientists interested in endothelial cell imaging in the zebrafish embryo.


Comparison of two beta-lactamase-producing strains of Streptococcus faecalis.
A second strain of enterococcus (PA) producing beta-lactamase (Bla+ phenotype) was compared with the previously reported Bla+ enterococcus, strain HH22. As with the original strain, there was a marked inoculum effect when PA was tested with penicillin, ampicillin, and piperacillin; no difference was noted with methicillin, cephalothin, imipenem, or vancomycin; the difference with ticarcillin was intermediate. High-level gentamicin resistance (Gmr) transferred from PA to an enterococcal recipient strain at a frequency approximately 100-fold lower than for HH22; all Gmr transconjugants from both strains were Bla+, but only PA showed linkage of Gmr and Bla+ with transfer of resistance to streptomycin, tetracycline, and chloramphenicol. EcoRI digestion of plasmid DNA from Gmr Bla+ transconjugants showed no similarities between the two strains. A 5.1-kilobase EcoRI Bla+-encoding fragment derived from HH22 was cloned into an Escherichia coli cloning vector and shown to hybridize to a 10.2-kilobase EcoRI fragment derived from PA; both fragments hybridized to an 840-base-pair staphylococcal Bla+ gene probe. These data indicate that the penicillinases are similar but encoded on different or differently arranged plasmids. The fact that both are transferable emphasizes the potential for this new streptococcal resistance determinant to disseminate.


Fine needle aspiration diagnosis of pilomatrixoma. A monomorphic population of basaloid cells with squamous differentiation not to be mistaken for carcinoma.
In a case of pilomatrixoma, an initial fine needle aspiration (FNA) biopsy of a skin lesion yielded only a monomorphic population of mitotically active basaloid cells with squamous differentiation, without anucleated ghost cells, chronic inflammatory cells or foreign-body giant cells. These findings suggested a metastatic squamous-cell carcinoma or a malignant tumor of adnexal skin. The correct diagnosis was made by repeat FNA, which yielded the basaloid cells and ghost cells characteristics of pilomatrixoma; the FNA diagnosis was confirmed by subsequent excisional biopsy. This case demonstrates that pilomatrixoma should be considered in the differential diagnosis of skin aspirates that contain a dominant population of basaloid cells. The differentiation of pilomatrixoma from malignant lesions of the head and neck region is discussed.


[Activation and regulation of nociceptive transient receptor potential (TRP) channels, TRPV1 and TRPA1].
TRP channels are well recognized for their contributions to sensory transduction, responding to a wide variety of stimuli including temperature, nociceptive stimuli, touch, osmolarity and pheromones. In particular, the involvement of TRP channels in nociception has been extensively studied following the cloning of the capsaicin receptor, TRPV1. Painful diabetic peripheral neuropathy is described as a superficial burning pain, and it is one of the most commonly encountered neuropathic pain syndromes in clinical practice. We found that hypoxic and high glucose conditions commonly observed in diabetes potentiate TRPV1 activity without affecting TRPV1 expression both in native rat sensory neurons and HEK293 cells expressing rat TRPV1. The potentiation seems to be caused by phosphorylation of the serine residues of TRPV1 by PKC. These data indicate that PKC-dependent potentiation of TRPV1 activities under hypoxia and hyperglycemia might be involved in early diabetic neuropathy. Mechanisms for the detection of alkaline pH by sensory neurons are not well understood, although it is well accepted that acidic pH monitoring can be attributed to several ion channels, including TRPV1 and ASICs. We found that alkaline pH activates TRPA1 and that the TRPA1 activation is involved in nociception, using Ca(2+)-imaging and patch-clamp methods. In addition, intracellular alkalization activated TRPA1 at the whole-cell level, and single-channel openings were observed in the inside-out configuration. Furthermore, intraplantar injection of ammonium chloride into the mouse hind paw caused pain-related behaviors, which were not observed in TRPA1-deficient mice. These results suggest that alkaline pH causes pain sensation through activation of TRPA1.


Student Perception of the Flipped Classroom in Nursing Education.
As nurse educators consider new pedagogy for teaching nursing, the students' views and perceptions must be considered. Many studies and commentaries have been written to date on the flipped classroom in nursing education, but few have used research methods to tease out student views of this teaching approach. The purpose of this study was to investigate student perceptions of the flipped classroom using a phenomenological approach. Students who had experienced traditional lecture instruction and the flipped classroom in the same semester were asked about their perceptions and comparisons of the two instructional methods. Qualitative analysis of fifteen individual interviews and two focus groups revealed seven themes, each encompassing a rich data set to inform nurse educators about student perceptions of the flipped classroom. This research informs educators on the effects of launching innovative teaching and learning strategies while considering the audience within the classroom.


Further characterization of Escherichia coli endonuclease V. Mechanism of recognition for deoxyinosine, deoxyuridine, and base mismatches in DNA.
Endonuclease V from Escherichia coli has a wide substrate spectrum. In addition to deoxyinosine-containing DNA, the enzyme cleaves DNA containing urea residues, AP sites, base mismatches, insertion/deletion mismatches, flaps, and pseudo-Y structures. The gene coding for the enzyme was identified to be orf 225 or nfi (endonuclease five). Using enzyme purified from an overproducing strain, the deoxyinosine- and mismatch-specific activities of endonuclease V was found to have different divalent metal requirements. The affinity of the enzyme is greater than 20-fold higher for DNA containing deoxyinosine than deoxynebularine or base mismatches. Under optimal cleavage conditions, endonuclease V forms two stable complexes with DNA containing deoxyinosine, but not with DNA containing base mismatches or deoxynebularine, suggesting that the 6-keto group of hypoxanthine in DNA is critical for stable interactions with the protein. The enzyme recognizes deoxyuridine in DNA but exhibits a much lower affinity to DNA containing deoxyuridine compared with DNA containing deoxyinosine. Interestingly, deoxyuridine-specific endonuclease activity of endonuclease V has a divalent metal requirement similar to the mismatch activity. A model for the mechanism of substrate recognition is proposed to explain these different activities.