Paradigm shift: should the elderly undergo propofol sedation for DBE? A prospective cohort study.
Evaluate the safety of propofol-assisted double balloon enteroscopy (DBE) in elderly patients against a younger cohort. Prospective cohort study. All patients undergoing DBE over a 30-month period were recruited at our tertiary centre. 215 procedures in 161 patients were performed. An age cut-off of 65 years and above was used to define those who were elderly. Patients were subcategorised into four groups: elderly or young undergoing DBE with propofol or conventional sedation (with midazolam±fentanyl). Patient demographics, comorbidities, procedural data, complications, diagnostic and therapeutic yield were compared. Cardiovascular disease and a higher American Society of Anaethesiologists (ASA) status were more prevalent in elderly patients undergoing DBE with propofol (p<0.05). Common indications for DBE were occult and overt obscure gastrointestinal bleeding and suspected Crohn's disease (elderly vs young: 50.7% vs 42.3%, 17.8% vs 12% and 19.2% vs 26.1%, respectively). Diagnostic yield was higher in elderly compared with young patients (75.3% vs 58.5%, p=0.016). The most common findings in elderly and young patients were angioectasia (30.1% and. 18.3%, respectively) and ulcers (17.8% and 9.2%, respectively), while therapeutic intervention rates were comparable (42.5% vs 32.4%, p=0.18). ASA status did not affect propofol dose (p=0.55) or procedure duration (p=0.31). Tolerance scores were favourable in those receiving propofol compared with conventional sedation (p<0.05). There was no difference in complications between the four groups (p=0.17). Compared with young patients, propofol-assisted DBE in the elderly is safe and has a high diagnostic yield.


Subjective judgements of synergistic risks: a cognitive reasoning perspective.
Mounting evidence that certain hazard combinations present synergistic risks for adverse outcomes, including violent crime, cancer, and species extinction, highlights the importance of understanding the risk attributable to combined hazards. However, previous studies indicate that individuals often misjudge synergistic risks as additive or sub-additive risks, and there is little research that explores the cognitive reasoning that may lead individuals to make such judgements. This study aims to fill this gap. Participants were asked to review several scenarios that described the risk magnitude presented by a combined hazard. They were required to judge whether each scenario was possible and to explain the reasoning that led to their judgement. The results show that many participants demonstrated an awareness of synergistic risk and that their reasoning was typically characterized by rudimentary knowledge of an underlying causal mechanism for the increased risk (e.g., a chemical reaction between drugs). Conversely, several participants adopted a line of reasoning that precluded the concept of synergistic risk. Many of these participants appeared to employ an additive model of risk, corresponding to the notion of 'adding' one hazard to another. Contrary to much previous research, we found little evidence to indicate that people tend to employ a sub-additive model of risk for combined hazards. Implications for future research and the improvement of risk communications concerning synergistic risks are discussed.


Bin1 ablation increases susceptibility to cancer during aging, particularly lung cancer.
Age is the major risk factor for cancer, but few genetic pathways that modify cancer incidence during aging have been described. Bin1 is a prototypic member of the BAR adapter gene family that functions in vesicle dynamics and nuclear processes. Bin1 limits oncogenesis and is often attenuated in human cancers, but its role in cancer suppression has yet to be evaluated fully in vivo. In the mouse, homozygous deletion of Bin1 causes developmental lethality, so to assess this role, we examined cancer incidence in mosaic null mice generated by a modified Cre-lox technology. During study of these animals, one notable phenotype was an extended period of female fecundity during aging, with mosaic null animals retaining reproductive capability until the age of 17.3 +/- 1.1 months. Through 1 year of age, cancer incidence was unaffected by Bin1 ablation; however, by 18 to 20 months of age, approximately 50% of mosaic mice presented with lung adenocarcinoma and approximately 10% with hepatocarcinoma. Aging mosaic mice also displayed a higher incidence of inflammation and/or premalignant lesions, especially in the heart and prostate. In mice where colon tumors were initiated by a ras-activating carcinogen, Bin1 ablation facilitated progression to more aggressive invasive status. In cases of human lung and colon cancers, immunohistochemical analyses evidenced frequent attenuation of Bin1 expression, paralleling observations in other solid tumors. Taken together, our findings highlight an important role for Bin1 as a negative modifier of inflammation and cancer susceptibility during aging.


Effects of Mindfulness-Based Stress Reduction on Depression in Adolescents and Young Adults: A Systematic Review and Meta-Analysis.
Background: Mindfulness as a positive mental health intervention approach has been increasingly applied to address depression in young people. This systematic review and meta-analysis evaluated the effects of mindfulness-based stress reduction (MBSR) in the treatment of depression among adolescents and young adults. Methods: Electronic databases and references in articles were searched. Randomized controlled trials (RCTs) evaluating MBSR and reporting outcomes for depressive symptoms among young people aged 12 to 25 years were included. Data extraction and risk of bias assessment were conducted by two reviewers independently. Hedges' g with a 95% confidence interval was calculated to represent intervention effect. Results: Eighteen RCTs featuring 2,042 participants were included in the meta-analysis. Relative to the control groups (e.g., no treatment, treatment as usual, or active control), MBSR had moderate effects in reducing depressive symptoms at the end of intervention (Hedges' g = -0.45). No statistically significant effects were found in follow-up (Hedges' g = -0.24) due to a lack of statistical power. Meta-regression found that the average treatment effect might be moderated by control condition, treatment duration, and participants' baseline depression. Conclusion: MBSR had moderate effects in reducing depression in young people at posttest. Future research is needed to assess the follow-up effects of MBSR on depressive symptoms among adolescents and young adults.


Executive functioning in highly talented soccer players.
Executive functions might be important for successful performance in sports, particularly in team sports requiring quick anticipation and adaptation to continuously changing situations in the field. The executive functions motor inhibition, attention and visuospatial working memory were examined in highly talented soccer players. Eighty-four highly talented youth soccer players (mean age 11.9), and forty-two age-matched amateur soccer players (mean age 11.8) in the age range 8 to 16 years performed a Stop Signal task (motor inhibition), the Attention Network Test (alerting, orienting, and executive attention) and a visuospatial working memory task. The highly talented soccer players followed the talent development program of the youth academy of a professional soccer club and played at the highest national soccer competition for their age. The amateur soccer players played at a regular soccer club in the same geographical region as the highly talented soccer players and play in a regular regional soccer competition. Group differences were tested using analyses of variance. The highly talented group showed superior motor inhibition as measured by stop signal reaction time (SSRT) on the Stop Signal task and a larger alerting effect on the Attention Network Test, indicating an enhanced ability to attain and maintain an alert state. No group differences were found for orienting and executive attention and visuospatial working memory. A logistic regression model with group (highly talented or amateur) as dependent variable and executive function measures that significantly distinguished between groups as predictors showed that these measures differentiated highly talented soccer players from amateur soccer players with 89% accuracy. Highly talented youth soccer players outperform youth amateur players on suppressing ongoing motor responses and on the ability to attain and maintain an alert state; both may be essential for success in soccer.


Rehabilitation of the atrophic maxilla with tilted implants: review of the literature.
We review the evidence-based literature on the use of tilted implants in the rehabilitation of patients with maxillary atrophy. Studies from 1999 to 2010 on patients with atrophic maxilla rehabilitated with tilted implants were reviewed. Clinical series with at least 10 patients rehabilitated using tilted implants and a follow-up of at least 12 months after prosthetic load were included. Case reports and studies with missing data were excluded. In each study the following was assessed: surgical technique, prosthesis type, timing of implant loading, success rate and marginal bone loss of tilted and axial implants, complications and patient satisfaction level. Thirteen studies were included, reporting a total of 782 tilted and 666 axial implants in 319 patients. Success rates went from 91.3% to 100% for axial implants and from 92.1% to 100% for tilted implants; radiographic marginal bone loss went from 0.4 mm to 0.92 mm in tilted implants and from 0.35 mm to 1.21 mm in axial implants. No statistically significant differences were found in any of the studies. No surgical complications and only minor prosthetic complications were reported. High patient satisfaction was found with all types of prosthesis (full-arch fixed, partial fixed and overdentures) placed over tilted implants. The literature on tilted implants shows that implants placed with this technique, both used alone and combined with axially placed implants, and rehabilitated with different prosthetic options have high success rates, minimal complications and high patient satisfaction. However, lack of homogeneity among studies and relatively short follow-up periods for most studies make necessary more studies.


Effect of dexmedetomidine on erythrocyte deformability during ischemia-reperfusion injury of liver in diabetic rats.
The aim of this study is to evaluate the effect of dexmedetomidine on erythrocyte deformability during IR injury of liver in diabetic rats. Twenty- eight Wistar Albino rats were included in the study after a 4 week streptozocin (65 mg/kg) treatment to observe the existence of diabetes. The animals were randomly assigned to one of the four experimental groups: GroupC and DC (sham-control group): The abdomen was dissected with a median laparotomy and the liver was collected. GroupDIR: The liver was collected after IR following the abdominal median laparotomy. GroupDIRD: The liver was collected after IR following the abdominal median laparotomy and 30 min of infusion of dexmedetomidine 100 µg/kg ip The deformability measurements were performed in erythrocyte suspensions containing Htc 5% in PBS buffer. The deformability index was significantly increased in diabetic rats, however it was similar in the GroupC and DIRD. It was significantly increased in the GroupDIR when compared to the GroupC, DIRD and DC. The relative resistance was increased in IR models. Erythrocyte deformability was damaged in rats having diabetes and IR injury. This injury might lead to further problems in microcirculation. It was shown that dexmedetomidine may be useful in enhancing the adverse effects of this injury (Tab. 1, Fig. 2, Ref. 41).


Cerebellar-motor dysfunction in schizophrenia and psychosis-risk: the importance of regional cerebellar analysis approaches.
Motor abnormalities in individuals with schizophrenia and those at-risk for psychosis are well documented. An accumulating body of work has also highlighted motor abnormalities related to cerebellar dysfunction in schizophrenia including eye-blink conditioning, timing, postural control, and motor learning. We have also recently found evidence for motor dysfunction in individuals at ultra high-risk for psychosis (1-3). This is particularly relevant as the cerebellum is thought to be central to the cognitive dysmetria model of schizophrenia, and these overt motor signs may point to more general cerebellar dysfunction in the etiology of psychotic disorders. While studies have provided evidence indicative of motor cerebellar dysfunction in at-risk populations and in schizophrenia, findings with respect to the cerebellum have been mixed. One factor potentially contributing to these mixed results is the whole-structure approach taken when investigating the cerebellum. In non-human primates, there are distinct closed-loop circuits between the cerebellum, thalamus, and brain with motor and non-motor cortical regions. Recent human neuroimaging has supported this finding and indicates that there is a cerebellar functional topography (4), and this information is being missed with whole-structure approaches. Here, we review cerebellar-motor dysfunction in individuals with schizophrenia and those at-risk for psychosis. We also discuss cerebellar abnormalities in psychosis, and the cerebellar functional topography. Because of the segregated functional regions of the cerebellum, we propose that it is important to look at the structure regionally in order to better understand its role in motor dysfunction in these populations. This is analogous to approaches taken with the basal ganglia, where each region is considered separately. Such an approach is necessary to better understand cerebellar pathophysiology on a macro-structural level with respect to the pathogenesis of psychosis.


Oleoresin chemistry mediates oviposition behavior and fecundity of a tree-killing bark beetle.
Many herbivores are sensitive to the secondary chemistry of their host plants. However, the influence of pine secondary chemicals (monoterpenes) on bark beetle fitness is poorly understood. We tested the hypothesis that the monoterpene composition of the phloem oleoresin of ponderosa pine, Pinus ponderosa var scopulorum, mediates rates of host acceptance, oviposition behavior, and fecundity of the western pine beetle, Dendroctonus brevicomis. We performed reciprocal rearing experiments, controlling for the monoterpene composition (chemotype) of host material. We tested the effects of two geographically interspersed host chemotypes on beetles with unknown (wild) and known (reared F(1)) chemical histories. Host chemotype and insect chemical history did not affect rates of acceptance of host material by female beetles. Insect chemical history affected egg gallery construction, and beetles constructed egg galleries that were on average 24.3% longer when reared in host material that was chemically similar to their natal host material. However, mean egg gallery lengths did not differ between host chemotypes. Insect chemical history also influenced fecundity: F(1) beetles produced 52.7% more offspring on average when reared in host material that was chemically similar to their natal host. Our experiments demonstrate that the chemical history of bark beetles mediates egg gallery construction and fecundity, but not host acceptance. This implicates chemical history as a more important factor than host chemotype in the oviposition behavior and fecundity of D. brevicomis.


Nucleotide diversity of the TNF gene region in an African village.
The wide variety of disease associations reported at the TNF locus raises the question of how much variation exists within a single population. To address this question, we sequenced the entire TNF gene in 72 chromosomes from healthy residents of a village in The Gambia, West Africa. We found 12 polymorphisms in 4393 nucleotides, of which five have not been previously described, giving an estimated nucleotide diversity (theta) of 5.6 x 10(-4). A significantly higher frequency of polymorphisms was found in the promoter region than in the coding region (8/1256 vs 0/882 nucleotides, P = 0.02). All polymorphisms with the exception of one rare allele were found to be present in Malawi, which is both geographically and genetically distant from The Gambia. Genotyping of 424 Gambian and 121 Malawian adults showed a significant frequency difference between the two populations for eight of the 12 polymorphisms, but the average fixation index across the variable sites was relatively low (F(ST) = 0.007). We conclude that, at the TNF locus, the nucleotide diversity found within a single African village is similar to the global value for human autosomal genes sampled across different continents.


Helicobacter pylori-induced REDD1 modulates Th17 cell responses that contribute to gastritis.
Regulated in development and DNA damage responses-1 (REDD1) is a conserved and ubiquitous protein, which is induced in response to multiple stimuli. However, the regulation, function and clinical relevance of REDD1 in Helicobacter pylori-associated gastritis are presently unknown. Immunohistochemistry, real-time PCR and Western blot analyses were performed to examine the levels of REDD1 in gastric samples from H. pylori-infected patients and mice. Gastric tissues from Redd1-/- and wildtype (WT, control) mice were examined for inflammation. Gastric epithelial cells (GECs), monocytes and T cells were isolated, stimulated and/or cultured for REDD1 regulation and functional assays. REDD1 was increased in gastric mucosa of H. pylori-infected patients and mice. H. pylori induced GECs to express REDD1 via the phosphorylated cytotoxin associated gene A (cagA) that activated MAPKp38 pathway to mediate NF-κB directly binding to REDD1 promoter. Human gastric REDD1 increased with the severity of gastritis, and mouse REDD1 from non-marrow chimera-derived cells promoted gastric inflammation that was characterized by the influx of MHCII+ monocytes. Importantly, gastric inflammation, MHCII+ monocyte infiltration, IL-23 and IL-17A were attenuated in Redd1-/- mice. Mechanistically, REDD1 in GECs regulated CXCL1 production, which attracted MHCII+ monocytes migration by CXCL1-CXCR2 axis. Then H. pylori induced MHCII+ monocytes to secrete IL-23, which favored IL-17A-producing CD4+ cell (Th17 cell) polarization, thereby contributing to the development of H. pylori-associated gastritis. The present study identifies a novel regulatory network involving REDD1, which collectively exert a pro-inflammatory effect within gastric microenvironment. Efforts to inhibit this REDD1-dependent pathway may prove valuable strategies in treating of H. pylori-associated gastritis.


Experimental demonstration of ray-rotation sheets.
We have built microstructured sheets that rotate, on transmission, the direction of light rays by an arbitrary, but fixed, angle around the sheet normal. These ray-rotation sheets comprise two pairs of confocal lenticular arrays. In addition to rotating the direction of transmitted light rays, our sheets also offset ray position sideways on the scale of the diameter of the lenticules. If this ray offset is sufficiently small so that it cannot be resolved, our ray-rotation sheets appear to perform generalized refraction.


Economic evaluation of interventions to reduce solar ultraviolet radiation (UVR) exposure among construction workers.
Solar ultraviolet radiation is one of the most common occupational carcinogens in Canada and is responsible for approximately 5,556 non-melanoma skin cancers per year. A large part of these cases are preventable by reducing solar ultraviolet radiation exposure. In this study, investigators estimated the potential economic impacts of different solar ultraviolet radiation reduction interventions among construction workers, as they are one of the largest at-risk occupational groups. Investigators performed an economic evaluation from a societal perspective, by comparing incremental costs in relation to incremental benefits achieved by two interventions-use of personal protective equipment by all exposed individuals and use of shade structure wherever and whenever feasible. Interventions costs were estimated for 2020-2050, and benefits with a 10-year delay, i.e., for the period 2030-2060. Economic evaluation estimates were reported by intervention costs, total costs of non-melanoma skin cancers cases averted, incremental cost per avert case, return on investment, and the break-even point. Various sensitivity analyses were undertaken with key parameters. Our findings indicate that if the rising trend of incidence continues, cases will be double in 2060, whereas by using personal protective equipment or shade structure, with the best-case scenario of full ultraviolet radiation removal, would result in 6,034 and 2,945 cases averted over 30 years, respectively. This translates into a total of $38.0 and $20.5 million of averted costs (all monetary values represented in 2017 Canadian dollars). Under this scenario investigators expect that by 2060, for every dollar invested in personal protective equipment and shade structures, $0.49 and $0.35 will be returned, respectively. Findings also suggested that under a conservative scenario, prevention of non-melanoma skin cancer cases by personal protective equipment and shade structures resulted in $5,812 and $7,355 incremental costs, respectively, over the 30-year period. This study provides important insights for decision makers about the potential impacts of solar ultraviolet radiation reduction interventions in the construction sector and other sectors with substantial outdoor work. Our estimates also can raise awareness of the importance of solar ultraviolet radiation reduction interventions.


Thalassaemia.
Thalassaemia is one of the most common genetic diseases worldwide, with at least 60,000 severely affected individuals born every year. Individuals originating from tropical and subtropical regions are most at risk. Disorders of haemoglobin synthesis (thalassaemia) and structure (eg, sickle-cell disease) were among the first molecular diseases to be identified, and have been investigated and characterised in detail over the past 40 years. Nevertheless, treatment of thalassaemia is still largely dependent on supportive care with blood transfusion and iron chelation. Since 1978, scientists and clinicians in this specialty have met regularly in an international effort to improve the management of thalassaemia, with the aim of increasing the expression of unaffected fetal genes to improve the deficiency in adult β-globin synthesis. In this Seminar we discuss important advances in the understanding of the molecular and cellular basis of normal and abnormal expression of globin genes. We will summarise new approaches to the development of tailored pharmacological agents to alter regulation of globin genes, the first trial of gene therapy for thalassaemia, and future prospects of cell therapy.


Hyperinsulinemia: a Cause of Obesity?
This perspective is motivated by the need to question dogma that does not work: that the problem is insulin resistance (IR). We highlight the need to investigate potential environmental obesogens and toxins. The prequel to severe metabolic disease includes three interacting components that are abnormal: (a) IR, (b) elevated lipids and (c) elevated basal insulin (HI). HI is more common than IR and is a significant independent predictor of diabetes. We hypothesize that (1) the initiating defect is HI that increases nutrient consumption and hyperlipidemia (HL); (2) the cause of HI may include food additives, environmental obesogens or toxins that have entered our food supply since 1980; and (3) HI is sustained by HL derived from increased adipose mass and leads to IR. We suggest that HI and HL are early indicators of metabolic dysfunction and treating and reversing these abnormalities may prevent the development of more serious metabolic disease.


Novel alanyl-tRNA synthetase 2 (AARS2) homozygous mutation in a consanguineous Chinese family with premature ovarian insufficiency.
To explore the candidate pathogenic gene in a premature ovarian insufficiency (POI) proband from a consanguineous marriage and detect the potential effects of mutation on cellular energy metabolism. Genetic and functional studies. Reproductive medicine center. A patient with POI, from a consanguineous family, and her family members were recruited from the Reproductive Center of the First Affiliated Hospital of Anhui Medical University. Whole exome sequencing (WES) was performed for the proband. Variation revealed by WES sequencing was validated by Sanger sequencing in her family. Sequencing data were combined with those of other sporadic cases listed in public databases to identify the causative gene. Rare homozygous nonsynonymous variants were identified and included in subsequent analysis. Metabolic analyzes were performed using Seahorse XFe96 analyzers to measure oxygen consumption and then obtain further results of ATP production and extracellular acidification rate. The differences in energy metabolism measurements between wild type and mutation were analyzed and compared. A novel alanyl-tRNA synthetase 2 (AARS2) homozygous mutation (NM_020745: exon2: c.337G>C: p. G113R) was identified in the aminoacylation region using WES. The mutation was highly conserved among species and predicted to be disease causing. AARS2 encodes mitochondrial alanyl-tRNA synthetase, which attaches alanine onto tRNA-ala. AARS2 mutations were previously reported in female leukodystrophy patients with POI. In mitochondrial stress tests, the ATP productions of the mutation group (3.58 ± 0.46 fmol/min/cell) was significantly lower than that of the wild type group (6.96 ± 1.56 fmol/min/cell). This is the first report of a homozygous pathogenic AARS2 mutation in POI. This mutation may lead to incorrect aminoacylation of tRNA, affect mitochondrial translation, and cause oxidative phosphorylation defects, which may be responsible for POI.


Dietary Docosahexaenoic Acid Improves Cognitive Function, Tissue Sparing, and Magnetic Resonance Imaging Indices of Edema and White Matter Injury in the Immature Rat after Traumatic Brain Injury.
Traumatic brain injury (TBI) is the leading cause of acquired neurologic disability in children. Specific therapies to treat acute TBI are lacking. Cognitive impairment from TBI may be blunted by decreasing inflammation and oxidative damage after injury. Docosahexaenoic acid (DHA) decreases cognitive impairment, oxidative stress, and white matter injury in adult rats after TBI. Effects of DHA on cognitive outcome, oxidative stress, and white matter injury in the developing rat after experimental TBI are unknown. We hypothesized that DHA would decrease early inflammatory markers and oxidative stress, and improve cognitive, imaging and histologic outcomes in rat pups after controlled cortical impact (CCI). CCI or sham surgery was delivered to 17 d old male rat pups exposed to DHA or standard diet for the duration of the experiments. DHA was introduced into the dam diet the day before CCI to allow timely DHA delivery to the pre-weanling pups. Inflammatory cytokines and nitrates/nitrites were measured in the injured brains at post-injury Day (PID) 1 and PID2. Morris water maze (MWM) testing was performed at PID41-PID47. T2-weighted and diffusion tensor imaging studies were obtained at PID12 and PID28. Tissue sparing was calculated histologically at PID3 and PID50. DHA did not adversely affect rat survival or weight gain. DHA acutely decreased oxidative stress and increased anti-inflammatory interleukin 10 in CCI brains. DHA improved MWM performance and lesion volume late after injury. At PID12, DHA decreased T2-imaging measures of cerebral edema and decreased radial diffusivity, an index of white matter injury. DHA improved short- and long-term neurologic outcomes after CCI in the rat pup. Given its favorable safety profile, DHA is a promising candidate therapy for pediatric TBI. Further studies are needed to explore neuroprotective mechanisms of DHA after developmental TBI.


Simultaneous temperature and sensitive two-species concentration measurements by single-shot CARS.
Simultaneous spatially and temporally resolved measurements of N(2) and O(2) mole fractions and of temperature are performed using coherent anti-Stokes Raman scattering (CARS). The CARS setup is used with the crossed-beam arrangement (BOXCARS) and nonresonant-background suppression. The technique employs two Stokes lasers, broadband and narrowband, in combination with a frequency-doubled Nd:YAG laser. Temperature and N(2) mole fractions are obtained by single-shot multiplex CARS spectra of N(2) using the broadband laser; O(2) mole fractions are deduced from a particular rovibrational Q-line of O(2) using the narrowband dye laser. The single-shot detectivity limit is better than 0.4% for oxygen at 2200 K and atmospheric pressure, i.e., 10(16) molecules x cm(-3). The capability of the technique for measuring 2-D probability density functions is demonstrated in the simple cases of an isothermal jet and a laminar premixed flame of air and ethylene. The experimental work reveals grave difficulties in using CARS for precise measurements of mole fractions: appreciable signals can be created very far from the geometrical focus; beam disruption by turbulence and the Stark effect cause large mole fraction measurement errors. These problems are discussed. Referencing the mole fraction of the second species by nitrogen mole fraction is demonstrated to be a solution for the turbulence effect in premixed flames.


The search for a human equivalent of the mouse T-locus - negative results from a study of HL-A types in spina bifida.
The spina bifida/anencephaly complex in man in some ways resembles the effects of T-locus mutants in the mouse. To test the hypothesis that these entities are genetically related, HL-A typing was performed in families with multiple cases of central nervous system abnormality. No evidence of linkage was found between HL-A type and the congenital malformation.


Germline mutations in the alternative pathway of complement predispose to HELLP syndrome.
HELLP (hemolysis, elevated liver enzymes, and low platelets) syndrome is a severe variant of hypertensive disorders of pregnancy affecting approximately 1% of all pregnancies, and has significant maternal and fetal morbidity. Previously, we showed that upregulation of the alternative pathway of complement (APC) plays a role in HELLP syndrome. We hypothesize that HELLP syndrome follows a 2-hit disease model similar to atypical hemolytic uremic syndrome (aHUS), requiring both genetic susceptibility and an environmental risk factor. Our objective was to perform a comparative analysis of the frequency of APC activation and germline mutations in affected women and to create a predictive model for identifying HELLP syndrome. Pregnant women with HELLP syndrome, and healthy controls after 23 weeks of gestation were recruited, along with aHUS and thrombotic thrombocytopenic purpura participants. We performed a functional assay, the mHam, and targeted genetic sequencing in all groups. Significantly more participants with rare germline mutations in APC genes were present in the HELLP cohort compared with controls (46% versus 8%, P = 0.01). In addition, significantly more HELLP participants were positive for the mHam when compared with controls (62% versus 16%, P = 0.009). Testing positive for both a germline mutation and the mHam was highly predictive for the diagnosis of HELLP syndrome. HELLP syndrome is characterized by both activation of the APC and frequent germline mutations in APC genes. Similar to aHUS, treatment via complement inhibition to mitigate maternal and fetal morbidity and mortality may be possible. National Heart Lung and Blood Institute grants T32HL007525 and R01HL133113.


Predisposing factors of restless legs syndrome in pregnancy.
The occurrence of restless legs syndrome in pregnancy is well known. However, the mechanism of this association is unclear. In this study, we aimed to identify the factors that predispose women to have restless legs syndrome during pregnancy. A total of 146 pregnant women were included in the study. Patients were asked questions regarding demographic characteristics, complications of pregnancy, medical therapy (vitamin and iron intake), sleep disorders, muscle cramps, and excessive daytime sleepiness. Electroneurography, routine blood biochemistry tests, complete blood count, and thyroid function tests were performed and vitamin B12, folic acid, serum iron, iron-binding capacity, ferritin, iron saturation, prolactin, estradiol, and progesterone were measured. Of the participants, 38 were diagnosed as having restless legs syndrome. In women with restless legs syndrome, additional medical problems, night cramps, and excessive daytime sleepiness were more frequent. In women without restless legs syndrome, serum hemoglobin levels were significantly higher and the use of supplemental iron or vitamins was greater. Among the women with restless legs syndrome, progesterone levels were slightly higher but this difference was not statistically significant. In summary, in this study, lower hemoglobin levels and supplementation deficits of iron and vitamins were found be the risk factors for restless legs syndrome in pregnancy.


A comparison of instrument sensitivity to negative symptom change.
Accurate assessment of negative symptom changes in schizophrenic patients is crucial to determining the efficacy of new treatments. The present study examined the sensitivity to change over the course of hospitalization in negative symptomatology assessed by the Scale for the Assessment of Negative Symptoms (SANS), the Negative Symptom Assessment (NSA), and an expanded version of the Brief Psychiatric Rating Scale (BPRS) in a sample of 60 schizophrenic patients. Symptoms were assessed when the patients were acutely ill and again when they were stabilized. Effect sizes were compared across all three rating scales. The retardation factor of the BPRS had a relatively small effect size (0.32). Effect size for the total NSA was 0.78 and ranged from 0.38 to 0.87 for the SANS. Individual factors had moderate to large effect sizes that ranged from 0.18 to 0.91 for both scales. Separate analyses were performed to calculate effect sizes for a five-factor version of the NSA and to examine only those symptoms specified in DSM-IV (alogia, affective flattening, and avolition). Effect sizes relatively comparable to those found for the entire SANS and NSA scales were found for the separate calculations. Results indicate that the addition of a negative symptom assessment instrument to research protocols increases the ability to detect changes in negative symptoms with substantially fewer subjects than would be required with the BPRS retardation factor alone.


Strategies to Optimize Adult Stem Cell Therapy for Tissue Regeneration.
Stem cell therapy aims to replace damaged or aged cells with healthy functioning cells in congenital defects, tissue injuries, autoimmune disorders, and neurogenic degenerative diseases. Among various types of stem cells, adult stem cells (i.e., tissue-specific stem cells) commit to becoming the functional cells from their tissue of origin. These cells are the most commonly used in cell-based therapy since they do not confer risk of teratomas, do not require fetal stem cell maneuvers and thus are free of ethical concerns, and they confer low immunogenicity (even if allogenous). The goal of this review is to summarize the current state of the art and advances in using stem cell therapy for tissue repair in solid organs. Here we address key factors in cell preparation, such as the source of adult stem cells, optimal cell types for implantation (universal mesenchymal stem cells vs. tissue-specific stem cells, or induced vs. non-induced stem cells), early or late passages of stem cells, stem cells with endogenous or exogenous growth factors, preconditioning of stem cells (hypoxia, growth factors, or conditioned medium), using various controlled release systems to deliver growth factors with hydrogels or microspheres to provide apposite interactions of stem cells and their niche. We also review several approaches of cell delivery that affect the outcomes of cell therapy, including the appropriate routes of cell administration (systemic, intravenous, or intraperitoneal vs. local administration), timing for cell therapy (immediate vs. a few days after injury), single injection of a large number of cells vs. multiple smaller injections, a single site for injection vs. multiple sites and use of rodents vs. larger animal models. Future directions of stem cell-based therapies are also discussed to guide potential clinical applications.


Extracorporeal lung support technologies - bridge to recovery and bridge to lung transplantation in adult patients: an evidence-based analysis.
For cases of acute respiratory distress syndrome (ARDS) and progressive chronic respiratory failure, the first choice or treatment is mechanical ventilation. For decades, this method has been used to support critically ill patients in respiratory failure. Despite its life-saving potential, however, several experimental and clinical studies have suggested that ventilator-induced lung injury can adversely affect the lungs and patient outcomes. Current opinion is that by reducing the pressure and volume of gas delivered to the lungs during mechanical ventilation, the stress applied to the lungs is eased, enabling them to rest and recover. In addition, mechanical ventilation may fail to provide adequate gas exchange, thus patients may suffer from severe hypoxia and hypercapnea. For these reasons, extracorporeal lung support technologies may play an important role in the clinical management of patients with lung failure, allowing not only the transfer of oxygen and carbon dioxide (CO(2)) but also buying the lungs the time needed to rest and heal. The objective of this analysis was to assess the effectiveness, safety, and cost-effectiveness of extracorporeal lung support technologies in the improvement of pulmonary gas exchange and the survival of adult patients with acute pulmonary failure and those with end-stage chronic progressive lung disease as a bridge to lung transplantation (LTx). The application of these technologies in primary graft dysfunction (PGD) after LTx is beyond the scope of this review and is not discussed. CLINICAL APPLICATIONS OF EXTRACORPOREAL LUNG SUPPORT: Extracorporeal lung support technologies [i.e., Interventional Lung Assist (ILA) and extracorporeal membrane oxygenation (ECMO)] have been advocated for use in the treatment of patients with respiratory failure. These techniques do not treat the underlying lung condition; rather, they improve gas exchange while enabling the implantation of a protective ventilation strategy to prevent further damage to the lung tissues imposed by the ventilator. As such, extracorporeal lung support technologies have been used in three major lung failure case types: As a bridge to recovery in acute lung failure - for patients with injured or diseased lungs to give their lungs time to heal and regain normal physiologic function.As a bridge to LTx - for patients with irreversible end stage lung disease requiring LTx.As a bridge to recovery after LTx - used as lung support for patients with PGD or severe hypoxemia. EX-VIVO LUNG PERFUSION AND ASSESSMENT: Recently, the evaluation and reconditioning of donor lungs ex-vivo has been introduced into clinical practice as a method of improving the rate of donor lung utilization. Generally, about 15% to 20% of donor lungs are suitable for LTx, but these figures may increase with the use of ex-vivo lung perfusion. The ex-vivo evaluation and reconditioning of donor lungs is currently performed at the Toronto General Hospital (TGH) and preliminary results have been encouraging (Personal communication, clinical expert, December 17, 2009). If its effectiveness is confirmed, the use of the technique could lead to further expansion of donor organ pools and improvements in post-LTx outcomes. EXTRACORPOREAL LUNG SUPPORT TECHNOLOGIES: ECMO: The ECMO system consists of a centrifugal pump, a membrane oxygenator, inlet and outlet cannulas, and tubing. The exchange of oxygen and CO(2) then takes place in the oxygenator, which delivers the reoxygenated blood back into one of the patient's veins or arteries. Additional ports may be added for haemodialysis or ultrafiltration. TWO DIFFERENT TECHNIQUES MAY BE USED TO INTRODUCE ECMO: venoarterial and venovenous. In the venoarterial technique, cannulation is through either the femoral artery and the femoral vein, or through the carotid artery and the internal jugular vein. In the venovenous technique cannulation is through both femoral veins or a femoral vein and internal jugular vein; one cannula acts as inflow or arterial line, and the other as an outflow or venous line. Venovenous ECMO will not provide adequate support if a patient has pulmonary hypertension or right heart failure. Problems associated with cannulation during the procedure include bleeding around the cannulation site and limb ischemia distal to the cannulation site. ILA: Interventional Lung Assist (ILA) is used to remove excess CO(2) from the blood of patients in respiratory failure. The system is characterized by a novel, low-resistance gas exchange device with a diffusion membrane composed of polymethylpentene (PMP) fibres. These fibres are woven into a complex configuration that maximizes the exchange of oxygen and CO(2) by simple diffusion. The system is also designed to operate without the help of an external pump, though one can be added if higher blood flow is required. The device is then applied across an arteriovenous shunt between the femoral artery and femoral vein. Depending on the size of the arterial cannula used and the mean systemic arterial pressure, a blood flow of up to 2.5 L/min can be achieved (up to 5.5 L/min with an external pump). The cannulation is performed after intravenous administration of heparin. Recently, the first commercially available extracorporeal membrane ventilator (NovaLung GmbH, Hechingen, Germany) was approved for clinical use by Health Canada for patients in respiratory failure. The system has been used in more than 2,000 patients with various indications in Europe, and was used for the first time in North America at the Toronto General Hospital in 2006. EVIDENCE-BASED ANALYSIS: The research questions addressed in this report are: Does ILA/ECMO facilitate gas exchange in the lungs of patients with severe respiratory failure?Does ILA/ECMO improve the survival rate of patients with respiratory failure caused by a range of underlying conditions including patients awaiting LTx?What are the possible serious adverse events associated with ILA/ECMO therapy?To address these questions, a systematic literature search was performed on September 28, 2009 using OVID MEDLINE, MEDLINE In-Process and Other Non-Indexed Citations, EMBASE, the Cumulative Index to Nursing & Allied Health Literature (CINAHL), the Cochrane Library, and the International Agency for Health Technology Assessment (INAHTA) for studies published from January 1, 2005 to September 28, 2008. Abstracts were reviewed by a single reviewer and, for those studies meeting the eligibility criteria, full-text articles were obtained. Reference lists were also examined for any additional relevant studies not identified through the search. Articles with an unknown eligibility were reviewed with a second clinical epidemiologist and then a group of epidemiologists until consensus was established. Studies in which ILA/ECMO was used as a bridge to recovery or bridge to LTxStudies containing information relevant to the effectiveness and safety of the procedureStudies including at least five patients Studies reporting the use of ILA/ECMO for inter-hospital transfers of critically ill patientsStudies reporting the use of ILA/ECMO in patients during or after LTxAnimal or laboratory studiesCase reports Reduction in partial pressure of CO(2)Correction of respiratory acidosisImprovement in partial pressure of oxygenImprovement in patient survivalFrequency and severity of adverse eventsThe search yielded 107 citations in Medline and 107 citations in EMBASE. After reviewing the information provided in the titles and abstracts, eight citations were found to meet the study inclusion criteria. One study was then excluded because of an overlap in the study population with a previous study. Reference checking did not produce any additional studies for inclusion. Seven case series studies, all conducted in Germany, were thus included in this review (see Table 1). Also included is the recently published CESAR trial, a multicentre RCT in the UK in which ECMO was compared with conventional intensive care management. The results of the CESAR trial were published when this review was initiated. In the absence of any other recent RCT on ECMO, the results of this trial were considered for this assessment and no further searches were conducted. A literature search was then conducted for application of ECMO as bridge to LTx patients (January, 1, 2005 to current). A total of 127 citations on this topic were identified and reviewed but none were found to have examined the use of ECMO as bridge to LTx. To grade the quality of evidence, the grading system formulated by the GRADE working group and adopted by MAS was applied. The GRADE system classifies the quality of a body of evidence as high, moderate, low, or very low according to four key elements: study design, study quality, consistency across studies, and directness. TRIALS ON ILA: Of the seven studies identified, six involved patients with ARDS caused by a range of underlying conditions; the seventh included only patients awaiting LTx. All studies reported the rate of gas exchange and respiratory mechanics before ILA and for up to 7 days of ILA therapy. Four studies reported the means and standard deviations of blood gas transfer and arterial blood pH, which were used for meta-analysis. Fischer et al. reported their first experience on the use of ILA as a bridge to LTx. In their study, 12 patients at high urgency status for LTx, who also had severe ventilation refractory hypercapnea and respiratory acidosis, were connected to ILA prior to LTx. Seven patients had a systemic infection or sepsis prior to ILA insertion. Six hours after initiation of ILA, the partial pressure of CO(2) in arterial blood significantly decreased (P < .05) and arterial blood pH significantly improved (P < .05) and remained stable for one week (last time point reported). (ABSTRACT TRUNCATED)


Constructing a yeast to express the largest cellulosome complex on the cell surface.
Cellulosomes, which are multienzyme complexes from anaerobic bacteria, are considered nature's finest cellulolytic machinery. Thus, constructing a cellulosome in an industrial yeast has long been a goal pursued by scientists. However, it remains highly challenging due to the size and complexity of cellulosomal genes. Here, we overcame the difficulties by synthesizing the Clostridium thermocellum scaffoldin gene (CipA) and the anchoring protein gene (OlpB) using advanced synthetic biology techniques. The engineered Kluyveromyces marxianus, a probiotic yeast, secreted a mixture of dockerin-fused fungal cellulases, including an endoglucanase (TrEgIII), exoglucanase (CBHII), β-glucosidase (NpaBGS), and cellulase boosters (TaLPMO and MtCDH). The confocal microscopy results confirmed the cell-surface display of OlpB-ScGPI and fluorescence-activated cell sorting analysis results revealed that almost 81% of yeast cells displayed OlpB-ScGPI. We have also demonstrated the cellulosome complex formation using purified and crude cellulosomal proteins. Native polyacrylamide gel electrophoresis and mass spectrometric analysis further confirmed the cellulosome complex formation. Our engineered cellulosome can accommodate up to 63 enzymes, whereas the largest engineered cellulosome reported thus far could accommodate only 12 enzymes and was expressed by a plasmid instead of chromosomal integration. Interestingly, CipA 2B9C (with two cellulose binding modules, CBM) released significantly higher quantities of reducing sugars compared with other CipA variants, thus confirming the importance of cohesin numbers and CBM domain on cellulosome complex. The engineered yeast host efficiently degraded cellulosic substrates and released 3.09 g/L and 8.61 g/L of ethanol from avicel and phosphoric acid-swollen cellulose, respectively, which is higher than any previously constructed yeast cellulosome.


Register based monitoring shows decreasing socioeconomic differences in Finnish perinatal health.
Several studies on differences in infant outcome by socioeconomic position have been done, but these have usually been based on ad hoc data linkages. The aim of this paper was to investigate whether socioeconomic differences in perinatal health in Finland could be regularly monitored using routinely collected data from one single register. Since October 1990, the Finnish Medical Birth Register (MBR) has included data on maternal occupation. A special computer program that converted the occupation name into an occupational code and into a socioeconomic position was prepared. Perinatal health was measured with five different indicators. The Finnish MBR data for years 1991 to 1999 (n=565 863 newborns) were used in the study. The study period was divided into three, three year periods to study time trends. An occupational code was derived for 95% of women, but it was not possible to define a socioeconomic position for 22% of women, including, for example, students and housewives (the group "Others"). For the rest, the data showed socioeconomic differences in all perinatal health indicators. Maternal smoking explained up to half of the excess risk for adverse perinatal outcome in the lowest socioeconomic group. The socioeconomic differences narrowed during the 1990s: infant outcome improved in the lowest socioeconomic group, but remained at the same level or even deteriorated in other groups. When comparing the lowest group with the highest group, the odds ratios (OR) adjusted for maternal background characteristics at least halved for prematurity (from 1.32 (95% confidence intervals 1.24 to 1.43) in 1991-1993 to 1.16 (1.08 to 1.25) in 1997-1999), for low birth weight (from 1.49 (1.36 to 1.63) to 1.25 (1.17 to 1.40)), and for perinatal mortality (from 1.79 (1.44 to 2.21) to 1.33 (1.07 to 1.66)). Social inequality in perinatal health outcomes exists in Finland, but seems to have diminished in the 1990s. These data showed that routinely collected birth register data provide a good source for studies on socioeconomic health differences in the perinatal period, but that uncertainty, mainly attributable to the large group of women with difficult to classify socioeconomic status, remains.


Investigations on the effects of dietary essential oils and different husbandry conditions on the gut ecology in piglets after weaning.
Essential oils (EO) are being considered as possible alternatives to in-feed antibiotic growth promoters in pig nutrition. The effects of an EO mixture consisting of limonene, eugenol and pinene (10.0, 2.0, and 4.8 mg/kg diet, resp.) on gut physiology and ecology were studied in piglets. The experiment was conducted at low (commercial farm) and high hygienic conditions (experimental farm), to elucidate interactions between EO supplementation and husbandry methods. Piglets were weaned at 28 days of age, when they were offered either a control diet (C) or C with EO. Four piglets were sacrificed in each group on day 29, 30, 33 and 39. Digesta from the third distal part of the small intestine and from the colon were sampled and analysed for pH, dry matter, lactic acid, short chain fatty acids and ammonia concentrations. Enterobacteria, enterococci, lactobacilli and yeast counts were obtained by plating. Genomic DNA was extracted from digesta and polymerase chain reaction-denaturing gradient gel electrophoresis was performed. Individual microbial communities were identified at each farm. Age affected the intestinal parameters. No effects of the EO with exception for a significant reduction in colon bacterial diversity at 39 days of age could be recorded at experimental farm.


Looking like Limulus? - Retinula axons and visual neuropils of the median and lateral eyes of scorpions.
Despite ongoing interest in the neurophysiology of visual systems in scorpions, aspects of their neuroanatomy have received little attention. Lately sets of neuroanatomical characters have contributed important arguments to the discussion of arthropod ground patterns and phylogeny. In various attempts to reconstruct phylogeny (from morphological, morphological + molecular, or molecular data) scorpions were placed either as basalmost Arachnida, or within Arachnida with changing sister-group relationships, or grouped with the extinct Eurypterida and Xiphosura inside the Merostomata. Thus, the position of scorpions is a key to understanding chelicerate evolution. To shed more light on this, the present study for the first time combines various techniques (Cobalt fills, DiI / DiO labelling, osmium-ethyl gallate procedure, and AMIRA 3D-reconstruction) to explore central projections and visual neuropils of median and lateral eyes in Euscorpius italicus (Herbst, 1800) and E. hadzii Di Caporiacco, 1950. Scorpion median eye retinula cells are linked to a first and a second visual neuropil, while some fibres additionally connect the median eyes with the arcuate body. The lateral eye retinula cells are linked to a first and a second visual neuropil as well, with the second neuropil being partly shared by projections from both eyes. Comparing these results to previous studies on the visual systems of scorpions and other chelicerates, we found striking similarities to the innervation pattern in Limulus polyphemus for both median and lateral eyes. This supports from a visual system point of view at least a phylogenetically basal position of Scorpiones in Arachnida, or even a close relationship to Xiphosura. In addition, we propose a ground pattern for the central projections of chelicerate median eyes.


Intramolecular hydride transfer onto arynes: redox-neutral and transition metal-free C(sp3)-H functionalization of amines.
Transition metal-free intramolecular hydride transfer onto arynes is reported for the first time. This unique transformation is utilized in redox-neutral intermolecular α-functionalization reactions of different tertiary amines, generating C(sp3)-C(sp3/sp2/sp) bonds in a single synthetic operation. Deuterium labeling studies support initial cleavage of the α-C-H bond via intramolecular 1,5-hydride transfer onto the aryne, which leads to activation of a range of integrated pronucleophiles and ultimately affords a new approach to cross-dehydrogenative coupling reactions which utilize aryne intermediates.


Identification of tissues and patterning events required for distinct steps in early migration of zebrafish primordial germ cells.
In many organisms, the primordial germ cells have to migrate from the position where they are specified towards the developing gonad where they generate gametes. Extensive studies of the migration of primordial germ cells in Drosophila, mouse, chick and Xenopus have identified somatic tissues important for this process and demonstrated a role for specific molecules in directing the cells towards their target. In zebrafish, a unique situation is found in that the primordial germ cells, as marked by expression of vasa mRNA, are specified in random positions relative to the future embryonic axis. Hence, the migrating cells have to navigate towards their destination from various starting positions that differ among individual embryos. Here, we present a detailed description of the migration of the primordial germ cells during the first 24 hours of wild-type zebrafish embryonic development. We define six distinct steps of migration bringing the primordial germ cells from their random positions before gastrulation to form two cell clusters on either side of the midline by the end of the first day of development. To obtain information on the origin of the positional cues provided to the germ cells by somatic tissues during their migration, we analyzed the migration pattern in mutants, including spadetail, swirl, chordino, floating head, cloche, knypek and no isthmus. In mutants with defects in axial structures, paraxial mesoderm or dorsoventral patterning, we find that certain steps of the migration process are specifically affected. We show that the paraxial mesoderm is important for providing proper anteroposterior information to the migrating primordial germ cells and that these cells can respond to changes in the global dorsoventral coordinates. In certain mutants, we observe accumulation of ectopic cells in different regions of the embryo. These ectopic cells can retain both morphological and molecular characteristics of primordial germ cells, suggesting that, in zebrafish at the early stages tested, the vasa-expressing cells are committed to the germ cell lineage.


Isotretinoin does not prolong QT intervals and QT dispersion in patients with severe acne: a surprising finding for a drug with numerous side effects.
Isotretinoin is a widely prescribed drug for the treatment of severe acne. Several adverse cardiac effects due to isotretinoin have been previously reported. However, no data exist on the effects of isotretinoin therapy on QT intervals. To investigate the effects of isotretinoin therapy on QT intervals and QT dispersion, and also to see if it is related to serum lipids, homocysteine and lipoprotein (a) or not. Forty-five patients with severe acne (mean age 21±6 years, range 14-38 years; 26 female) were included in the study. Twelve-lead surface electrocardiograms (ECGs) were acquired at three stages: before therapy and at the ends of the first and sixth months of 0.8 mg/kg/day of isotretinoin therapy. Serum levels of triglycerides, total cholesterol, low density lipoprotein cholesterol, high density lipoprotein cholesterol, very low density lipoprotein cholesterol, homocysteine and lipoprotein (a) were also measured at the day of ECG recordings. Minimum and maximum QT intervals were measured and QT dispersion was calculated. Mean heart rates were similar throughout the isotretinoin therapy. Serum levels of lipids, homocysteine and lipoprotein (a) all increased significantly at the end of the first month and remained significantly elevated at the end of sixth month (P is less than 0.05 for both stages). QT intervals and QT dispersion did not differ significantly throughout the six months of isotretinoin therapy (P is greater than 0.05). In patients with severe acne, six months of 0.8 mg/kg/day of isotretinoin therapy neither prolongs QT interval, nor increases QT dispersion. This effect is not related to blood lipids, homocysteine or lipoprotein (a) levels. Our findings indicate that from the point of polymorphic ventricular tachycardia risk, 0.8 mg/kg/day of isotretinoin therapy is a safe choice in acne treatment.


Identification and cardiotropic actions of brain/gut-derived tachykinin-related peptides (TRPs) from the American lobster Homarus americanus.
Two tachykinin-related peptides (TRPs) are known in decapods, APSGFLGMRamide and TPSGFLGMRamide. The former peptide appears to be ubiquitously conserved in members of this taxon, while the latter has been suggested to be a genus (Cancer)- or infraorder (Brachyura)-specific isoform. Here, we characterized a cDNA from the American lobster Homarus americanus (infraorder Astacidea) that encodes both TRPs: six copies of APSGFLGMRamide and one of TPSGFLGMRamide. Mass spectral analyses of the H. americanus supraoesophageal ganglion (brain) and commissural ganglia confirmed the presence of both peptides in these neural tissues; both isoforms were also detected in the midgut. Physiological experiments showed that both APSGFLGMRamide and TPSGFLGMRamide are cardioactive in H. americanus, eliciting identical increases in both heart contraction frequency and amplitude. Collectively, our data represent the first genetic confirmation of TRPs in H. americanus and of TPSGFLGMRamide in any species, demonstrate that TPSGFLGMRamide is not restricted to brachyurans, and show that both this peptide and APSGFLGMRamide are brain-gut isoforms, the first peptides thus far confirmed to possess this dual tissue distribution in H. americanus. Our data also suggest a possible role for TRPs in modulating the output of the lobster heart.


Adverse events following vaccination in premature infants.
The aims of this study were to study the frequency, severity and types of adverse reactions following DPT/Hib (diphtheria and tetanus toxoids and pertussis/Haemophilus influenzae type B conjugate) immunization in very preterm infants and to identify possible risk factors. Case notes of 45 preterm babies vaccinated in the neonatal intensive care unit between January 1993 and December 1998 were studied retrospectively. Birthweight, gestational age, duration of ventilation, oxygen dependency, timing of vaccination, weight, corrected gestation at vaccination and apparent adverse effects were noted. Apparent adverse events were noted in 17 of 45 (37.8%) babies: 9 (20%) had major events, i.e. apnoea, bradycardia or desaturations, and 8 (17.8%) had minor events, i.e. increased oxygen requirements, temperature instability, poor handling and feed intolerance. Babies with major events were significantly younger (p < 0.05), had a lower postmenstrual age (p < 0.05) and weighed less (p < 0.05) at the time of vaccination compared with babies without major events. No differences in the mean birthweight, gestational age, duration of ventilation or oxygen dependency were found between the two groups. Age at vaccination of 70 days or less was significantly associated with increased risk (p < 0.01). Of 27 babies vaccinated at 70 days or less, 9 (33.3%) developed major events compared with none when vaccinated over 70 d. Vaccine-related cardiorespiratory events are relatively common in preterm babies. Problems were much more common if vaccine is administered at or before 70 d. These babies should therefore be monitored postvaccination. Further prospective studies are needed to clarify whether delaying vaccination offers protection against these adverse events.


Enhanced dopamine efflux in the amygdala by a predictive, but not a non-predictive, stimulus: facilitation by prior repeated D-amphetamine.
Extracellular levels of dopamine within the amygdala were monitored using in vivo microdialysis during performance of an appetitive Pavlovian conditioning task in sensitized rats and unsensitized controls. Animals received exposure either to D-amphetamine or to vehicle for seven consecutive days (2 mg/kg/day, i.p.) in the home cage. Training began following a further seven injection-free days. Animals were exposed to two session types: during conditioning sessions, a stimulus (tone or light) immediately preceded sucrose pellet delivery. During control sessions, the alternative stimulus was also presented, but not in temporal proximity to an otherwise identical schedule of pellet delivery. There was a total of three alternating presentations of each session type during training. Sensitization enhanced Pavlovian conditioned approach behaviour to the stimulus predictive of imminent pellet delivery, and was without effect upon approach behaviours either to the food pellets themselves or to the control stimulus. Extracellular levels of dopamine within the amygdala were assessed during the fourth conditioning and control sessions. Mesoamygdaloid dopamine efflux increased significantly during the conditioning test session, but not during the control session, and this dopaminergic response was more marked in rats with prior repeated D-amphetamine experience. Hence, these results add to evidence suggesting a role for amygdaloid dopamine in appetitive Pavlovian conditioning, and in the facilitation of associative learning following prior experience of D-amphetamine.


Expression of heat shock protein 70 is altered by age and diet at the level of transcription.
Because heat shock proteins have been shown to play a critical role in protecting cells from hyperthermia and other types of physiological stresses, it was of interest to determine what effect age and caloric restriction have on the ability of cells to regulate the expression of heat shock protein 70 (hsp70), the most prominent and most evolutionarily conserved of the heat shock proteins. Caloric restriction is the only experimental manipulation known to retard aging and increase survival of mammals. The ability of hepatocytes isolated from young/adult (4- to 7-month-old) and old (22- to 28-month-old) male Fischer F344 rats fed ad libitum or a caloric restriction diet (60% of the content of the ad libitum diet) to express hsp70 was determined after a mild heat shock (42.5 degrees C for 30 min). We found that the induction of hsp70 synthesis and mRNA levels by heat shock was 40 to 50% lower in hepatocytes isolated from old rats than in hepatocytes isolated from young rats. Using in situ hybridization, we found that essentially all hepatocytes from the young/adult and old rats expressed hsp70 in response to heat shock; therefore, the age-related decrease in the induction of hsp70 expression was not due to an age-related accumulation of cells that do not respond to heat shock. Measurements of hsp70 mRNA stability and hsp70 transcription demonstrated that the age-related decline in hsp70 expression arose from a decline in hsp70 transcription. Interestingly, the age-related decline in the induction of hsp70 expression was reversed by caloric restriction; e.g., the induction of hsp70 synthesis, mRNA levels, and nuclear transcription were significantly higher in hepatocytes isolated from old rats fed the caloric restricted diet than in hepatocytes isolated from old rats fed ad libitum. The levels of the heat shock transcription factor in nuclear extracts isolated from heat-shocked hepatocytes were measured in a gel shift assay. Binding of the heat shock transcription factor to the heat shock element decreased with age and was significantly higher in hepatocyte extracts isolated from old rats fed the caloric restriction diet than in those from old rats fed ad libitum. Thus, our study demonstrates that the ability of hepatocytes to respond to hyperthermia and express hsp70 decreases significantly with age and that this decrease occurs at the transcriptional level. In addition, caloric restriction, which retards aging, reversed the age-related decline in the induction of hsp70 transcription in hepatocytes.


Reciprocal relations between parents' physical discipline and children's externalizing behavior during middle childhood and adolescence.
Using data from two long-term longitudinal projects, we investigated reciprocal relations between maternal reports of physical discipline and teacher and self-ratings of child externalizing behavior, accounting for continuity in both discipline and externalizing over time. In Study 1, which followed a community sample of 562 boys and girls from age 6 to 9, high levels of physical discipline in a given year predicted high levels of externalizing behavior in the next year, and externalizing behavior in a given year predicted high levels of physical discipline in the next year. In Study 2, which followed an independent sample of 290 lower income, higher risk boys from age 10 to 15, mother-reported physical discipline in a given year predicted child ratings of antisocial behavior in the next year, but child antisocial behavior in a given year did not predict parents' use of physical discipline in the next year. In neither sample was there evidence that associations between physical discipline and child externalizing changed as the child aged, and findings were not moderated by gender, race, socioeconomic status, or the severity of the physical discipline. Implications for the reciprocal nature of the socialization process and the risks associated with physical discipline are discussed.


The amount of ergonomics and user involvement in 151 design processes.
Ergonomics, usability and user-centered design are terms that are well known among designers. Yet, products often seem to fail to meet the users' needs, resulting in a gap between expected and experienced usability. To understand the possible causes of this gap the actions taken by the designer during the design process are studied in this paper. This can show whether and how certain actions influence the user-friendliness of the design products. The aim of this research was to understand whether ergonomic principles and methods are included in the design process, whether users are involved in this process and whether the experience of the designer (in ergonomics/user involvement) has an effect on the end product usability. In this study the design processes of 151 tangible products of students in design were analyzed. It showed that in 75% of the cases some ergonomic principles were applied. User involvement was performed in only 1/3 of the design cases. Hardly any correlation was found between the designers' experience in ergonomic principles and the way they applied it and no correlations were found between the designers' experience in user involvement and the users' involvement in the design process.


Enzyme systems for biodegradation of polychlorinated dibenzo-p-dioxins.
The angular dioxygenase, cytochrome P450, lignin peroxidase, and dehalogenase are known as dioxin-metabolizing enzymes. All of these enzymes have metal ions in their active centers, and the enzyme systems except for peroxidase have each distinct electron transport chain. Although the enzymatic properties of the angular dioxygenase, lignin peroxidase, and cytochrome P450 have been studied well, the information about dehalogenase is much less than other enzyme systems due to its instability under the aerobic conditions. However, this enzyme system appears to be quite promising from the viewpoint of practical use for bioremediation, because dehalogenases are capable of degradation of polychlorinated dibenzo-p-dioxins (PCDDs) with more than four chlorine substituents, whereas the other three enzyme systems prefer low-chlorinated PCDDs. On the other hand, protein engineering of angular dioxygenase, lignin peroxidase, and cytochrome P450 based on their tertiary structures has great potential to generate highly efficient dioxin-metabolizing enzymes. Actually, we successfully generated 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-metabolizing enzyme by site-directed mutagenesis of cytochrome P450. We hope that recombinant microorganisms harboring genetically engineered dioxin-metabolizing enzymes will be used for bioremediation of soil contaminated with PCDDs and polychlorinated dibenzofurans in the near future.


Coordinated control of replication and transcription by a SAPK protects genomic integrity.
Upon environmental changes or extracellular signals, cells are subjected to marked changes in gene expression. Dealing with high levels of transcription during replication is critical to prevent collisions between the transcription and replication pathways and avoid recombination events. In response to osmostress, hundreds of stress-responsive genes are rapidly induced by the stress-activated protein kinase (SAPK) Hog1 (ref. 6), even during S phase. Here we show in Saccharomyces cerevisae that a single signalling molecule, Hog1, coordinates both replication and transcription upon osmostress. Hog1 interacts with and phosphorylates Mrc1, a component of the replication complex. Phosphorylation occurs at different sites to those targeted by Mec1 upon DNA damage. Mrc1 phosphorylation by Hog1 delays early and late origin firing by preventing Cdc45 loading, as well as slowing down replication-complex progression. Regulation of Mrc1 by Hog1 is completely independent of Mec1 and Rad53. Cells carrying a non-phosphorylatable allele of MRC1 (mrc1(3A)) do not delay replication upon stress and show a marked increase in transcription-associated recombination, genomic instability and Rad52 foci. In contrast, mrc1(3A) induces Rad53 and survival in the presence of hydroxyurea or methyl methanesulphonate. Therefore, Hog1 and Mrc1 define a novel S-phase checkpoint independent of the DNA-damage checkpoint that permits eukaryotic cells to prevent conflicts between DNA replication and transcription, which would otherwise lead to genomic instability when both phenomena are temporally coincident.


Phylogenetic distribution of branched RNA-linked multicopy single-stranded DNA among natural isolates of Escherichia coli.
Multicopy single-stranded DNA (msDNA), a branched DNA-RNA molecule, has been shown in Escherichia coli B and clinical strain Cl-1 to be synthesized by reverse transcriptase. We report that 13% of the strains of the ECOR collection, a sample of 72 E. coli isolates representing the breadth of genetic variation of the species, produce msDNA. Three of the four major subspecific groups include msDNA-producing strains. Screening of 25 isolates that are genetically related to msDNA-producing clinical strains uncovered 22 additional msDNA-producing strains. A phylogenetic tree based on allelic variation detected electrophoretically at 20 enzyme-encoding loci revealed two major clusters and several deep branches composed of strains that synthesize msDNA. Although E. coli K-12 does not harbor msDNA, other closely related strains of the K-12 family do. The results support the hypothesis that msDNA-synthesizing systems, including reverse transcriptase genes, were acquired recently and independently in different lineages of E. coli.


Structures of ABCB10, a human ATP-binding cassette transporter in apo- and nucleotide-bound states.
ABCB10 is one of the three ATP-binding cassette (ABC) transporters found in the inner membrane of mitochondria. In mammals ABCB10 is essential for erythropoiesis, and for protection of mitochondria against oxidative stress. ABCB10 is therefore a potential therapeutic target for diseases in which increased mitochondrial reactive oxygen species production and oxidative stress play a major role. The crystal structure of apo-ABCB10 shows a classic exporter fold ABC transporter structure, in an open-inwards conformation, ready to bind the substrate or nucleotide from the inner mitochondrial matrix or membrane. Unexpectedly, however, ABCB10 adopts an open-inwards conformation when complexed with nonhydrolysable ATP analogs, in contrast to other transporter structures which adopt an open-outwards conformation in complex with ATP. The three complexes of ABCB10/ATP analogs reported here showed varying degrees of opening of the transport substrate binding site, indicating that in this conformation there is some flexibility between the two halves of the protein. These structures suggest that the observed plasticity, together with a portal between two helices in the transmembrane region of ABCB10, assist transport substrate entry into the substrate binding cavity. These structures indicate that ABC transporters may exist in an open-inwards conformation when nucleotide is bound. We discuss ways in which this observation can be aligned with the current views on mechanisms of ABC transporters.


Retrospective Analysis of 91 Kaposi's Sarcoma Cases: A Single-Center Experience and Review of the Literature.
Kaposi's sarcoma (KS) is a multifocal angioproliferative tumor involving primarily the skin. The aim of this study was to describe the clinical, demographic, histopathological characteristics, treatment modalities, and outcome of 91 KS patients, and compare them with other contemporary research. Medical records of 91 KS patients followed between January 2005 and September 2017 were evaluated retrospectively. Most of our patients were male (male-to-female ratio was 4.05). The median age at diagnosis was 69 years (range, 6-93 years). The duration of the lesions varied between 3 and 25 years. The lower extremities were the most commonly involved area (51.6%). Of the 91 patients, classic type KS was seen in 75 patients. Radiotherapy was used successfully in approximately half of our patients. Recurrence was observed in approximately one third of the patients. All KS patients in this study except 1 were classic KS. The clinical and demographic characteristics of our patients were compatible with the previous literature suggesting that KS is a tumor that tends to be limited to the skin. Close follow-up of patients is important to monitor for recurrence. This is the largest report from Turkey to date.


Estimating Rn-induced lung cancer in the United States.
The proportion of lung cancer deaths attributable to Rn among residents of single-family homes in the U.S. (approximately 70% of the housing stock) is estimated using the log-normal distribution of Rn concentrations proposed by Nero et al. (1986) and the risk model developed by the National Academy of Sciences' BEIR IV Committee. The risk model, together with the exposure distribution, predicts that approximately 14% of lung cancer deaths among such residents (about 13,300 deaths per year, or 10% of all U.S. lung cancer deaths) may be due to indoor Rn exposure. The 95% confidence interval is 7%-25%, or approximately 6600 to 24,000 lung cancer deaths. These estimated attributable risks due to Rn are similar for males and females and for smokers and nonsmokers, but higher baseline risks of lung cancer result in much larger absolute numbers of Rn-attributable cancers among males (approximately 9000) and among smokers (approximately 11,000). Because of the apparent skewness of the exposure distribution, most of the contribution to the attributable risks arises from exposure rates below 148 Bq m-3 (4 pCi L-1), i.e., below the EPA "action level." As a result, if all exposure rates that exceed 148 Bq m-3 (approximately 8% of homes) were eliminated, the models predict that the total annual lung cancer burden in the U.S. would drop by 4-5%, or by about 3800 lung cancer deaths, in contrast to a maximum reduction of 14% if all indoor Rn exposure above the 1st percentile were eliminated.


Probing the correlation between insulin activity and structural stability through introduction of the rigid A6-A11 bond.
The development of fast-acting and highly stable insulin analogues is challenging. Insulin undergoes structural transitions essential for binding and activation of the insulin receptor (IR), but these conformational changes can also affect insulin stability. Previously, we substituted the insulin A6-A11 cystine with a rigid, non-reducible C=C linkage ("dicarba" linkage). A cis-alkene permitted the conformational flexibility of the A-chain N-terminal helix necessary for high-affinity IR binding, resulting in surprisingly rapid activity in vivo Here, we show that, unlike the rapidly acting LysB28ProB29 insulin analogue (KP insulin), cis-dicarba insulin is not inherently monomeric. We also show that cis-dicarba KP insulin lowers blood glucose levels even more rapidly than KP insulin, suggesting that an inability to oligomerize is not responsible for the observed rapid activity onset of cis-dicarba analogues. Although rapid-acting, neither dicarba species is stable, as assessed by fibrillation and thermodynamics assays. MALDI analyses and molecular dynamics simulations of cis-dicarba insulin revealed a previously unidentified role of the A6-A11 linkage in insulin conformational dynamics. By controlling the conformational flexibility of the insulin B-chain helix, this linkage affects overall insulin structural stability. This effect is independent of its regulation of the A-chain N-terminal helix flexibility necessary for IR engagement. We conclude that high-affinity IR binding, rapid in vivo activity, and insulin stability can be regulated by the specific conformational arrangement of the A6-A11 linkage. This detailed understanding of insulin's structural dynamics may aid in the future design of rapid-acting insulin analogues with improved stability.


Enhanced bioavailability and efficiency of curcumin for the treatment of asthma by its formulation in solid lipid nanoparticles.
Curcumin has shown considerable pharmacological activity, including anti-inflammatory, but its poor bioavailability and rapid metabolization have limited its application. The purpose of the present study was to formulate curcumin-solid lipid nanoparticles (curcumin-SLNs) to improve its therapeutic efficacy in an ovalbumin (OVA)-induced allergic rat model of asthma. A solvent injection method was used to prepare the curcumin-SLNs. Physiochemical properties of curcumin-SLNs were characterized, and release experiments were performed in vitro. The pharmacokinetics in tissue distribution was studied in mice, and the therapeutic effect of the formulation was evaluated in the model. The prepared formulation showed an average size of 190 nm with a zeta potential value of -20.7 mV and 75% drug entrapment efficiency. X-ray diffraction analysis revealed the amorphous nature of the encapsulated curcumin. The release profile of curcumin-SLNs was an initial burst followed by sustained release. The curcumin concentrations in plasma suspension were significantly higher than those obtained with curcumin alone. Following administration of the curcumin-SLNs, all the tissue concentrations of curcumin increased, especially in lung and liver. In the animal model of asthma, curcumin-SLNs effectively suppressed airway hyperresponsiveness and inflammatory cell infiltration and also significantly inhibited the expression of T-helper-2-type cytokines, such as interleukin-4 and interleukin-13, in bronchoalveolar lavage fluid compared to the asthma group and curcumin-treated group. These observations implied that curcumin-SLNs could be a promising candidate for asthma therapy.


Hypothalamic UDP Increases in Obesity and Promotes Feeding via P2Y6-Dependent Activation of AgRP Neurons.
Activation of orexigenic AgRP-expressing neurons in the arcuate nucleus of the hypothalamus potently promotes feeding, thus defining new regulators of AgRP neuron activity could uncover potential novel targets for obesity treatment. Here, we demonstrate that AgRP neurons express the purinergic receptor 6 (P2Y6), which is activated by uridine-diphosphate (UDP). In vivo, UDP induces ERK phosphorylation and cFos expression in AgRP neurons and promotes action potential firing of these neurons in brain slice recordings. Consequently, central application of UDP promotes feeding, and this response is abrogated upon pharmacologic or genetic inhibition of P2Y6 as well as upon pharmacogenetic inhibition of AgRP neuron activity. In obese animals, hypothalamic UDP content is elevated as a consequence of increased circulating uridine concentrations. Collectively, these experiments reveal a potential regulatory pathway in obesity, where peripheral uridine increases hypothalamic UDP concentrations, which in turn can promote feeding via PY6-dependent activation of AgRP neurons.


Comparison of three inhaled non-steroidal anti-inflammatory drugs on the airway response to sodium metabisulphite and adenosine 5'-monophosphate challenge in asthma.
Non-steroidal anti-inflammatory drugs (NSAIDs) are used to assess the role of prostaglandins in asthma but their effects on bronchoconstrictor challenges have been inconsistent. The effects of three nebulised nonsteroidal anti-inflammatory drugs on the airway response to inhaled sodium metabisulphite (MBS) and adenosine 5'-monophosphate (AMP) were compared in the same asthmatic subjects to see whether contractile prostaglandins were involved in MBS or AMP induced bronchoconstriction. A possible protective effect of the osmolarity or pH of the inhaled solutions was also assessed. Two double blind placebo controlled studies were carried out. In study 1, 15 non-aspirin sensitive patients with mild asthma attended on four occasions and inhaled 5 ml of lysine aspirin (L-aspirin) 900 mg, indomethacin 50 mg, sodium salicylate 800 mg, or saline 20 minutes before an inhaled MBS challenge. On four further occasions 14 of the patients inhaled the same solutions followed by an inhaled AMP challenge. In study 2, 10 of the patients attended on four additional occasions and inhaled 5 ml of 0.9%, 3%, 10%, or 9.5% saline with indomethacin 50 mg 20 minutes before an inhaled MBS challenge. In study 1 inhaled lysine aspirin had a similar effect on MBS and AMP induced bronchoconstriction, increasing the provocative dose causing a 20% fall in FEV1 (PD20) by 1.29 (95% CI 0.54 to 2.03) and 1.23 (95% CI 0.53 to 1.93) doubling doses, respectively. Indomethacin increased the MBS PD20 and AMP PD20 by 0.64 (95% CI -0.1 to 1.38) and 0.99 (95% CI 0.29 to 1.69) doubling doses, respectively. Sodium salicylate had no significant effect on either challenge. The two solutions causing most inhibition were the most acidic and the most alkaline. In study 2 inhaled 9.5% saline with indomethacin (osmolarity 3005 mOsm/kg) increased the MBS PD20 by 1.1 doubling doses (95% CI 0.2 to 2.0) compared with only 0.09 (95% CI -0.83 to 1.0) and 0.04 (95% CI -0.88 to 0.95) doubling doses with 3% saline (918 mOsm/kg) and 10% saline (2994 mOsm/ kg), respectively. Inhaled L-aspirin and indomethacin have broadly similar protective effects against MBS and AMP induced bronchoconstriction in the doses given, although the effect of indomethacin on MBS was not quite statistically significant. The osmolarity and pH of the solutions did not appear to be important determinants of the response. The effect of L-aspirin and indomethacin is likely to be the result of cyclooxygenase inhibition reducing the production of contractile prostaglandins during MBS and AMP challenge.


Racial microaggressions in everyday life: implications for clinical practice.
Racial microaggressions are brief and commonplace daily verbal, behavioral, or environmental indignities, whether intentional or unintentional, that communicate hostile, derogatory, or negative racial slights and insults toward people of color. Perpetrators of microaggressions are often unaware that they engage in such communications when they interact with racial/ethnic minorities. A taxonomy of racial microaggressions in everyday life was created through a review of the social psychological literature on aversive racism, from formulations regarding the manifestation and impact of everyday racism, and from reading numerous personal narratives of counselors (both White and those of color) on their racial/cultural awakening. Microaggressions seem to appear in three forms: microassault, microinsult, and microinvalidation. Almost all interracial encounters are prone to microaggressions; this article uses the White counselor--client of color counseling dyad to illustrate how they impair the development of a therapeutic alliance. Suggestions regarding education and training and research in the helping professions are discussed.


Larotrectinib: A Targeted Therapy for Solid Tumors.
Although neurotrophic tyrosine receptor kinase (NTRK) gene fusions are not common in most cancers, they are present in more than 90% of some rare tumors. The U.S. Food and Drug Administration has approved larotrectinib for patients with NTRK gene fusion-positive cancers that meet certain criteria. With ongoing advancements in tumor sequencing, it is anticipated that cancer treatment will be determined by genetic variants rather than by cancer type in the future. This article provides an overview of larotrectinib, a targeted therapy. This article reviews clinical trial results and highlights implications for oncology nurses caring for patients taking larotrectinib. Larotrectinib is an effective treatment option for some patients with NTRK gene fusion-positive cancers. Oncology nurses are key to educating patients on dosing, administration, side effects, and precautions.


Multi-omic signature of body weight change: results from a population-based cohort study.
Excess body weight is a major risk factor for cardiometabolic diseases. The complex molecular mechanisms of body weight change-induced metabolic perturbations are not fully understood. Specifically, in-depth molecular characterization of long-term body weight change in the general population is lacking. Here, we pursued a multi-omic approach to comprehensively study metabolic consequences of body weight change during a seven-year follow-up in a large prospective study. We used data from the population-based Cooperative Health Research in the Region of Augsburg (KORA) S4/F4 cohort. At follow-up (F4), two-platform serum metabolomics and whole blood gene expression measurements were obtained for 1,631 and 689 participants, respectively. Using weighted correlation network analysis, omics data were clustered into modules of closely connected molecules, followed by the formation of a partial correlation network from the modules. Association of the omics modules with previous annual percentage weight change was then determined using linear models. In addition, we performed pathway enrichment analyses, stability analyses, and assessed the relation of the omics modules with clinical traits. Four metabolite and two gene expression modules were significantly and stably associated with body weight change (P-values ranging from 1.9 × 10(-4) to 1.2 × 10(-24)). The four metabolite modules covered major branches of metabolism, with VLDL, LDL and large HDL subclasses, triglycerides, branched-chain amino acids and markers of energy metabolism among the main representative molecules. One gene expression module suggests a role of weight change in red blood cell development. The other gene expression module largely overlaps with the lipid-leukocyte (LL) module previously reported to interact with serum metabolites, for which we identify additional co-expressed genes. The omics modules were interrelated and showed cross-sectional associations with clinical traits. Moreover, weight gain and weight loss showed largely opposing associations with the omics modules. Long-term weight change in the general population globally associates with serum metabolite concentrations. An integrated metabolomics and transcriptomics approach improved the understanding of molecular mechanisms underlying the association of weight gain with changes in lipid and amino acid metabolism, insulin sensitivity, mitochondrial function as well as blood cell development and function.


Fine-mapping and candidate gene analysis of bovine spinal muscular atrophy.
Bovine spinal muscular atrophy (SMA), an autosomal recessive neurodegenerative disease, has been mapped at moderate resolution to the distal part of Chromosome 24. In this article we confirm this location and fine-map the SMA locus to an interval of approximately 0.8 cM at the very distal end of BTA24. Despite remarkable similarity to human SMA, the causative gene SMN can be excluded in bovine SMA. However, the interval where the disease now has been mapped contains BCL2, like SMN an antiapoptotic factor, and shown to bind to SMN. Moreover, knockout mice lacking the BCL2 gene show rapid motor neuron degeneration with early postnatal onset, as observed in bovine SMA. A comparative cattle/human map of the distal end of BTA24, based on the emerging bovine genome sequencing data, shows conserved synteny to HSA18 with hints of a segmental duplication and pericentric inversion just after the last available bovine marker DIK4971. This synteny lets us conclude that SMA is in immediate vicinity of the telomere. Candidate gene analysis of BCL2, however, excludes most of this gene, except its promoter region, and draws attention to the neighboring gene VPS4B, part of the endosomal protein-sorting machinery ESCRT-III which is involved in several neurodegenerative diseases.


Glycemic Control for Patients With Type 2 Diabetes Mellitus: Our Evolving Faith in the Face of Evidence.
We sought to determine the concordance between the accumulating evidence about the impact of tight versus less tight glycemic control in patients with type 2 diabetes mellitus since the publication of UKPDS (UK Prospective Diabetes Study) in 1998 until 2015 with the views about that evidence published in journal articles and practice guidelines. We searched in top general medicine and specialty journals for articles referring to glycemic control appearing between 2006 and 2015 and identified the latest practice guidelines. To summarize the evidence, we included all published systematic reviews and meta-analyses of contemporary randomized trials of glycemic control measuring patient-important microvascular and macrovascular outcomes, and completed a meta-analysis of their follow-up extensions. We identified 16 guidelines and 328 statements. The body of evidence produced estimates warranting moderate confidence. This evidence reported no significant impact of tight glycemic control on the risk of dialysis/transplantation/renal death, blindness, or neuropathy. In the past decade, however, most published statements (77%-100%) and guidelines (95%) unequivocally endorsed benefit. There is also no significant effect on all-cause mortality, cardiovascular mortality, or stroke; however, there is a consistent 15% relative-risk reduction of nonfatal myocardial infarction. Between 2006 and 2008, most statements (47%-83%) endorsed the benefit; after 2008 (ACCORD), only a minority (21%-36%) did. Discordance exists between the research evidence and academic and clinical policy statements about the value of tight glycemic control to reduce micro- and macrovascular complications. This discordance may distort priorities in the research and practice agendas designed to improve the lives of patients with type 2 diabetes mellitus.


Longitudinal changes in cortical thickness associated with normal aging.
Imaging studies of anatomic changes in regional gray matter volumes and cortical thickness have documented age effects in many brain regions, but the majority of such studies have been cross-sectional investigations of individuals studied at a single point in time. In this study, using serial imaging assessments of participants in the Baltimore Longitudinal Study of Aging (BLSA), we investigate longitudinal changes in cortical thickness during aging in a cohort of 66 older adults (mean age 68.78; sd. 6.6; range 60-84 at baseline) without dementia. We used the Cortical Reconstruction Using Implicit Surface Evolution CRUISE suite of algorithms to automatically generate a reconstruction of the cortical surface and identified twenty gyral based regions of interest per hemisphere. Using mixed effects regression, we investigated longitudinal changes in these regions over a mean follow-up interval of 8 years. The main finding in this study is that age-related decline in cortical thickness is widespread, but shows an anterior-posterior gradient with frontal and parietal regions, in general, exhibiting greater rates of decline than temporal and occipital. There were fewer regions in the right hemisphere showing statistically significant age-associated longitudinal decreases in mean cortical thickness. Males showed greater rates of decline in the middle frontal, inferior parietal, parahippocampal, postcentral, and superior temporal gyri in the left hemisphere, right precuneus and bilaterally in the superior parietal and cingulate regions. Significant nonlinear changes over time were observed in the postcentral, precentral, and orbitofrontal gyri on the left and inferior parietal, cingulate, and orbitofrontal gyri on the right.


Genome-wide SNP discovery in walnut with an AGSNP pipeline updated for SNP discovery in allogamous organisms.
A genome-wide set of single nucleotide polymorphisms (SNPs) is a valuable resource in genetic research and breeding and is usually developed by re-sequencing a genome. If a genome sequence is not available, an alternative strategy must be used. We previously reported the development of a pipeline (AGSNP) for genome-wide SNP discovery in coding sequences and other single-copy DNA without a complete genome sequence in self-pollinating (autogamous) plants. Here we updated this pipeline for SNP discovery in outcrossing (allogamous) species and demonstrated its efficacy in SNP discovery in walnut (Juglans regia L.). The first step in the original implementation of the AGSNP pipeline was the construction of a reference sequence and the identification of single-copy sequences in it. To identify single-copy sequences, multiple genome equivalents of short SOLiD reads of another individual were mapped to shallow genome coverage of long Sanger or Roche 454 reads making up the reference sequence. The relative depth of SOLiD reads was used to filter out repeated sequences from single-copy sequences in the reference sequence. The second step was a search for SNPs between SOLiD reads and the reference sequence. Polymorphism within the mapped SOLiD reads would have precluded SNP discovery; hence both individuals had to be homozygous. The AGSNP pipeline was updated here for using SOLiD or other type of short reads of a heterozygous individual for these two principal steps. A total of 32.6X walnut genome equivalents of SOLiD reads of vegetatively propagated walnut scion cultivar 'Chandler' were mapped to 48,661 'Chandler' bacterial artificial chromosome (BAC) end sequences (BESs) produced by Sanger sequencing during the construction of a walnut physical map. A total of 22,799 putative SNPs were initially identified. A total of 6,000 Infinium II type SNPs evenly distributed along the walnut physical map were selected for the construction of an Infinium BeadChip, which was used to genotype a walnut mapping population having 'Chandler' as one of the parents. Genotyping results were used to adjust the filtering parameters of the updated AGSNP pipeline. With the adjusted filtering criteria, 69.6% of SNPs discovered with the updated pipeline were real and could be mapped on the walnut genetic map. A total of 13,439 SNPs were discovered by BES re-sequencing. BESs harboring SNPs were in 677 FPC contigs covering 98% of the physical map of the walnut genome. The updated AGSNP pipeline is a versatile SNP discovery tool for a high-throughput, genome-wide SNP discovery in both autogamous and allogamous species. With this pipeline, a large set of SNPs were identified in a single walnut cultivar.