Disrupting surfaces of nef required for downregulation of CD4 and for enhancement of virion infectivity attenuates simian immunodeficiency virus replication in vivo.
The multifunctional simian and human immunodeficiency virus (SIV and HIV) Nef proteins are important for virulence. We studied the importance of selected Nef functions using an SIV Nef with mutations in two regions that are required for CD4 downregulation. This Nef mutant is defective for downregulating CD4 and, in addition, for enhancing SIV infectivity and induction of SIV replication from infected quiescent peripheral blood mononuclear cells, but not for other known functions, including downregulation of class I major histocompatibility complex (MHC) cell surface expression. Replication of SIV containing this Nef variant in rhesus monkeys was attenuated early during infection. Subsequent increases in viral load coincided with selection of reversions and second-site compensatory changes in Nef. Our results indicate that the surfaces of Nef that mediate CD4 downregulation and the enhancement of virion infectivity are critical for SIV replication in vivo. Furthermore, these findings indicate that class I MHC downregulation by Nef is not sufficient for SIV virulence early in infection.


What underscored successful malaria elimination in Palestine 100 years ago? Effective Education.
Transmission of malaria by anopheline mosquitoes had been established by 1897, and in 1922, the first start of a successful national malaria elimination campaign began. Until then, only malaria control had been considered anywhere as a feasible project, such malaria control having been conducted primarily through larval source management. From 1922 onwards, in Palestine, by ensuring the breeding sites remained destroyed continuously over years and years, malaria elimination was eventually achieved. However, in order to achieve such continuous destruction, transmission of the disease had to be imaginatively and sensitively explained to all the inhabitants who thereupon willingly accepted the task of ensuring the breeding sites remained destroyed. Without that education, the inhabitants would not have provided the continuous work required, and Palestine would have remained in its severe malarious state.


Health information technology adoption in U.S. acute care hospitals.
Previous studies show that the healthcare industry lags behind many other economic sectors in the adoption of information technology. The purpose of this study is to understand differences in structural characteristics between providers that do and that do not adopt Health Information Technology (HIT) applications. Publicly available secondary data were used from three sources: American Hospital Association (AHA) annual survey, Healthcare Information and Management Systems Society (HIMSS) analytics annual survey, and Healthcare Cost and Utilization Project (HCUP) Nationwide Inpatient Sample (NIS) databases. Fifty-two information technologies were grouped into three clusters: clinical, administrative, and strategic decision making ITs. Negative binomial regression was applied with adoption of technology as the dependent variables and eight organizational and contextual factors as the independent variables. Hospitals adopt a relatively larger proportion of administrative information technology as compared to clinical and strategic IT. Large size, urban location and HMO penetration were found to be the most influential hospital characteristics that positively affect information technology adoption. There are still considerable variations in the adoption of information technology across hospitals and in the type of technology adopted. Organizational factors appear to be more influential than market factors when it comes to information technology adoption. The future research may examine whether the Electronic Health Record (EHR) Incentive Program in 2011 would increase the information technology uses in hospitals as it provides financial incentives for HER adoptions and uses among providers.


Exhaled Propofol Concentrations Correlate With Plasma and Brain Tissue Concentrations in Rats.
Propofol can be measured in exhaled gas. Exhaled and plasma propofol concentrations correlate well, but the relationship with tissue concentrations remains unknown. We thus evaluated the relationship between exhaled, plasma, and various tissue propofol concentrations. Because the drug acts in the brain, we focused on the relationship between exhaled and brain tissue propofol concentrations. Thirty-six male Sprague-Dawley rats were anesthetized with propofol, ketamine, and rocuronium for 6 hours. Animals were randomly assigned to propofol infusions at 20, 40, or 60 mg·kg·h (n = 12 per group). Exhaled propofol concentrations were measured at 15-minute intervals by multicapillary column-ion mobility spectrometry. Arterial blood samples, 110 µL each, were collected 15, 30, and 45 minutes, and 1, 2, 4, and 6 hours after the propofol infusion started. Propofol concentrations were measured in brain, lung, liver, kidney, muscle, and fat tissue after 6 hours. The last exhaled and plasma concentrations were used for linear regression analyses with tissue concentrations. The correlation of exhaled versus plasma concentrations (R = 0.71) was comparable to the correlation of exhaled versus brain tissue concentrations (R = 0.75) at the end of the study. In contrast, correlations between plasma and lung and between lung and exhaled propofol concentrations were poor. Less than a part-per-thousand of propofol was exhaled over 6 hours. Exhaled propofol concentrations correlate reasonably well with brain tissue and plasma concentrations in rats, and may thus be useful to estimate anesthetic drug effect. The equilibration between plasma propofol and exhaled gas is apparently independent of lung tissue concentration. Only a tiny fraction of administered propofol is eliminated via the lungs, and exhaled quantities thus have negligible influence on plasma concentrations.


Effect of NSAIDs on Recovery From Acute Skeletal Muscle Injury: A Systematic Review and Meta-analysis.
There is debate as to whether the use of nonsteroidal anti-inflammatory drugs (NSAIDs) is beneficial after acute skeletal muscle injury. Some studies have suggested that NSAID use may be detrimental to injured muscle. To determine whether NSAID use affects recovery from skeletal muscle injury as assessed by strength loss, soreness, and/or blood creatine kinase level. Systematic review and meta-analysis. An extensive systematic review was completed searching 16 databases (eg, PubMed, Cochrane Library, EMBASE). Inclusion criteria were (1) acute injury to skeletal muscle, (2) use of a control condition, (3) certainty of the NSAID dose administered, and (4) use of 1 or more of the 3 desired outcome measures. A total of 5343 study reports were screened, of which 41 studies were deemed suitable for inclusion. The standardized mean difference was used as the effect size (ES) and was calculated such that a positive ES indicated NSAID efficacy. Meta-analyses were run using a random-effects model. For all studies, time points after injury, and injury markers combined, NSAID use was found to elicit a small to medium, significant decrease in the markers of injury (overall ES = +0.34; P = .0001). Because heterogeneity in study ES was apparent (ie, Q- df = 52.4, P = .000005; I2 = 57%), subgroup meta-analyses and meta-regressions were run in an attempt to explain the heterogeneity. In human studies, study ESs were higher when lower body muscles were injured ( P = .045). In animal studies, study ESs were lower with longer NSAID administration durations ( P = .023) and at longer follow-up times after injury ( P = .010). Overall, our analysis supports NSAID use for reducing strength loss, soreness, and blood creatine kinase level after an acute muscle injury, at least for humans and in the short term. Additional research is required to determine why NSAID use appears to be more effective when lower-body muscles in humans are injured. It would also be important to determine why NSAID use appears detrimental at later times after injury in animals but not humans.


Early total care in polytrauma patient with floating shoulder and occlusion of bilateral subclavian artery: A case report.
Fracture in polytrauma patients have higher risk of morbidity and mortality compared to those found in monotrauma patients. The ideal approach of orthopaedic injuries is to perform definitive fixation of all fractures in one single procedure, an approach known as Early Total Care (ETC). A patient presented with history of being struck by heavy material forklift. He complained of pain in the shoulder and difficulty in breathing. The patient was diagnosed with polytrauma ISS Score 25, which consists of floating shoulder, closed fracture of multiple ribs, open hematopneumothorax, and thrombosis of subclavian artery. The following procedures were performed: debridement, ORIF plate and screw of right clavicle, ORIF lag screw of right glenoid scapula, thoracotomy segmental lobectomy, bypass of bilateral carotid artery to bilateral brachial artery. An early surgical treatment is paramount in the management of this patient, with the aim of restoring the joint and blood flow to the distal part of the hand, which will allow for early motion and more effective physiotherapy for the patient'srecovery. Early definitive fracture fixation in ETC in recommended in for stable patients and those who falls under the category of borderline and unstable patients who responds well to resuscitation. ETC allows for early mobilization and therapies for the patient. Early total care is an appropriate choice of treatment for polytrauma patients presenting with floating shoulder.


"Constructive" vs. "Destructive" deviance in adolescent health-related behaviors.
Previous literature suggests that the adolescent at risk to engage in substance use and other negative health-related behaviors is deviant in a negative sense (i.e., rebellious, antisocial, and alienated from traditional institutions). However, some researchers have distinguished between two types of deviance-a true autonomy and independence that is more positive and constructive, and a reactant "anticonformity" that is more negative and destructive. The current study assessed the roles of both constructive and destructive deviance in adolescent cigarette smoking and positive health-related behaviors. Adolescents who were constructively deviant engaged in higher levels of health-protective behaviors. Moreover, constructive deviance was an independent predictor of both cigarette smoking and positive health behaviors over and above the effects of traditional negative deviance indicators. These data suggest that constructive deviance is not a competing model to more traditional notions, but that it is an additional possible pathway into adolescent positive and negative health behaviors.


Inter-BAN Interference Mitigation: A Correlated Equilibrium Perspective.
A Body Area Network (BAN) is a wireless network of wearable or implantable computing devices. A BAN typically consists of several miniaturized radio-enabled body sensor/actuator that communicate with a single coordinator. Medical applications usually impose stringent constraints on the BAN operational reliability, quality of service, and power consumption. However, as there is no coordination among multiple co-located BANs, cross-interference could make achieving these objectives a challenging problem. Assuming Time Division Multiple Access (TDMA) at each BAN, this paper investigates the ability of regret matching based transmission scheduling algorithm to ease the impact of inter-BAN interference. This scheduling algorithm uses pattern of past interference for implicit coordination between different BAN transmissions. Simulation results demonstrate potential benefits of the proposed scheme for inter-BAN interference mitigation.


[Inhibitory effects of codonopsis pilosula polysaccharides on the deterioration of impaired phagocytosis of alveolar macrophage induced by fine particulate matter in chronic obstructive pulmonary disease mice].
To investigate the inhibitory effects of codonopsis pilosula polysaccharides (CPP) on the deterioration of impaired phagocytosis of alveolar macrophage (AM) induced by fine particulate matter with a mean aerodynamic diameter ≤2.5 μm (PM2.5) in chronic obstructive pulmonary disease (COPD) mice. Sixty BALB/c male mice were randomly divided into control group, COPD group, PM2.5 group, PM2.5 COPD group, CPP COPD group and CPP+ PM2.5 COPD group. COPD mice were established using exposure of cigarette smoking. Meanwhile PM2.5 group, PM2.5 COPD group and CPP+ PM2.5 COPD group were exposed to PM2.5 (770 μg/m(3)) for 90 days. CPP COPD group and CPP+ PM2.5 COPD group were fed with CPP (300 mg/kg) for 90 days whilst other groups were fed with isovolumetric saline. After the models were established, mice peak inspiratory flow (PIF) and peak expiratory flow (PEF) were measured by noninvasive body plethysmograph and lung histopathology and mean linear intercept (MLI) were observed. AMs were isolated from lung tissue by discontinuous density gradient centrifugation. Mean fluorescence intensity (MFI) and the ability of AM phagocytosing flurescein isothiocyanate-labeled Escherichia coli (FITC-E.coli) (AM%) were detected by flow cytometry. Total antioxidative capacity (TAC) was measured by O-phenanthroline colorimetry. Malondialdehyde (MDA) was measured by thiobarbiturieacid colorimetry and glutathione peroxidase (GSH-PX) by improved Hafeman colorimetry. MFI in control group, COPD group, PM2.5 group, PM2.5 COPD group, CPP COPD group and CPP+ PM2.5 COPD group were 10 267±1 358, 4 817±399, 8 469±240, 3 176±501, 5 886±516 and 4 067±453. AM% in each group were (69.0±5.4)%, (30.7±3.0)%, (51.5±2.4)%, (20.4±3.5)%, (38.7±2.6)% and (28.7±4.3)%. MFI and AM% in COPD group and PM2.5 group were decreased than those in control group while those in PM2.5 COPD group were lower than in COPD group (all P<0.01). Comparing to COPD group and PM2.5 COPD group respectively, MFI and AM% in CPP COPD group and CPP + PM2.5 COPD group were increased (all P<0.01). TAC and GSH-PX in each group were (17.99±0.09), (6.83±0.36), (13.84±1.12), (3.61±0.29), (8.80±0.26), (5.43±0.30) U/mg protein and (84.3±5.7), (46.5±2.6), (62.0±2.2), (32.4±3.8), (53.4±4.0), (42.4±4.0) U/mg. TAC and GSH-PX in COPD group and PM2.5 group were lower than those from control group while those from PM2.5 COPD group were decreased than in COPD group (all P<0.01). Comparing to COPD group and PM2.5 COPD group, TAC and GSH-PX in CPP COPD group and CPP+ PM2.5 COPD group were increased respectively (all P<0.01). MDA in each group were (1.74±0.37), (2.73±0.22), (2.01±0.13), (3.55±0.33), (2.22±0.28) and (2.72±0.44) nmol/mg protein. MDA in COPD group and PM2.5 group were higher than that from control group while that from PM2.5 COPD group was higher than in COPD group; MDA in CPP COPD group and CPP+ PM2.5 COPD group were respectively decreased than those in COPD group and PM2.5 COPD group (all P<0.05). Positive correlations were existed between MFI, AM% and TAC, GSH-PX, while negative correlations were existed between MFI, AM% and MDA in COPD group, PM2.5 group, PM2.5 COPD group, CPP COPD group and CPP+ PM2.5 COPD group. PM2.5 further impaired the defective phagocytosing capacity of AM and exacerbated oxidative stress in COPD mice. CPP can inhibit these effects. The protection of CPP may be closely related to its antioxidative effects.


A study of the effectiveness of family planning clinics in the Philippines.
A study of 190 Philippine family planning clinics revealed that certain clinic and physician characteristics were significantly and positively associated with clinic effectiveness, defined in terms of the mean number of monthly acceptors of family planning and the efficacy of the methods selected. Clinics with a high degree of effectiveness were more likely to be staffed by physicians who wanted three or fewer children, who relied more extensively on the IUD (intrauterine device) than physicians in less effective clinics, and who derived more satisfaction from working in family planning than physicians in less effective clinics. Autonomous clinics were more effective than integrated clinics, and clinics staffed by physicians trained by institutions with more experience in family planning were more effective than those staffed by physicians trained by institutions with less experience in this field. The findings regarding the following hypotheses were in the predicted direction: Clinic effectiveness will be greater in clinics staffed by female rather than by male physicians, in urban rather than in rural clinics, in clinics staffed by physicians with a low rather than a high degree of religiosity, and in clinics staffed by physicians with a high rather than a low degree of concern about population growth. The differences in clinic effectiveness, however, in these instances, were not significant.


[Neuroendocrine tumors of the gastrointestinal tract. Prospective analysis of 29 patients and review of the literature].
Neuroendocrine tumors are rare and can be observed in different gastrointestinal organs. The diagnosis often is made by incidence like in carcinoids of the appendix vermiformis, or special clinical symptoms may develop due to tumor growth. 29 consecutive patients were evaluated prospectively during 30 months. In 21 of them, the first diagnosis was obtained after an operative procedure. In 3 cases, the tumor was found incidentally during another oncological procedure. In 4 patients, the histological examination of biopsy specimens had shown an undifferentiated adenocarcinoma. In 8 of the patients, recurrent surgery was necessary because of recurrency of the disease. The mean observation time of all patients was 10.6 months. 6 of 29 patients died on disease-related symptoms. The results are discussed based on the literature with special regard to organ-related characteristics of different gastrointestinal neuroendocrine tumors.


Induction of Gsk3β-β-TrCP interaction is required for late phase stabilization of β-catenin in canonical Wnt signaling.
A pivotal step in canonical Wnt signaling is Wnt-induced β-catenin stabilization. In the absence of Wnt, β-catenin is targeted for β-transducin repeats-containing proteins (β-TrCP)-mediated degradation due to phosphorylation by glycogen synthase kinase 3 (Gsk3). How canonical Wnt signaling regulates Gsk3 to inhibit β-catenin proteolysis remains largely elusive. This study reveals novel key molecular events in Wnt signaling: induction of Gsk3β ubiquitination and Gsk3β-β-TrCP binding. We found that Wnt stimulation induced prolonged monoubiquitination of Gsk3β and Gsk3β-β-TrCP interaction. Monoubiquitination did not cause Gsk3β degradation nor affects its enzymatic activity. Rather, increased monoubiquitination of Gsk3β/Gsk3β-β-TrCP association suppressed β-catenin recruitment of β-TrCP, leading to long-term inhibition of β-catenin ubiquitination and degradation.


Crystal structures of high affinity human T-cell receptors bound to peptide major histocompatibility complex reveal native diagonal binding geometry.
Naturally selected T-cell receptors (TCRs) are characterised by low binding affinities, typically in the range 1-100 microM. Crystal structures of syngeneic TCRs bound to peptide major histocompatibility complex (pMHC) antigens exhibit a conserved mode of binding characterised by a distinct diagonal binding geometry, with poor shape complementarity (SC) between receptor and ligand. Here, we report the structures of three in vitro affinity enhanced TCRs that recognise the pMHC tumour epitope NY-ESO(157-165) (SLLMWITQC). These crystal structures reveal that the docking mode for the high affinity TCRs is identical to that reported for the parental wild-type TCR, with only subtle changes in the mutated complementarity determining regions (CDRs) that form contacts with pMHC; both CDR2 and CDR3 mutations act synergistically to improve the overall affinity. Comparison of free and bound TCR structures for both wild-type and a CDR3 mutant reveal an induced fit mechanism arising from restructuring of CDR3 loops which allows better peptide binding. Overall, an increased interface area, improved SC and additional H-bonding interactions are observed, accounting for the increase in affinity. Most notably, there is a marked increase in the SC for the central methionine and tryptophan peptide motif over the native TCR.


A rest-activity biomarker to predict response to SSRIs in major depressive disorder.
Most adults with Major Depressive Disorder (MDD) will not experience a remission with the first antidepressant trial. No practical biomarkers presently exist to predict responsiveness to antidepressants. Herein we report pilot data for a rest-activity biomarker of antidepressant response. Fifty-eight medication-free adults with MDD underwent a week-long collection of actigraphic data before beginning a 9 week open label trial of fluoxetine, coupled with blinded randomized assignment to eszopiclone/placebo. Depression severity was repeatedly measured with the Hamilton Rating Scale for Depression (HRSD). Baseline actigraphic data was analyzed with functional data analysis to create smoothed 24-h curves of activity. The time of the lowest point of activity (the bathyphase) was calculated for each patient, as well the mean difference between bedtime and the bathyphase (BBD). At the end of treatment, patients were characterized as treatment responders (50% reduction in HRSD) or non-responders, and receiver operating curves were calculated to find the optimal cut point of the BBD for prediction of treatment response. The best cut point for BBD was at 260.2 min, resulting in an effect size of 1.45, and with a positive predictive value of 0.75 and a negative predictive value of 0.88. We conclude that actigraphically-determined measures of rest-activity patterns show promise as potential biomarker predictors of antidepressant response. However, this conclusion is based upon a small number of patients who received only one choice of antidepressant, for a single trial. Replication with a larger sample is needed.


Extraction platform evaluations: a comparison of AutoMate Express™, EZ1® Advanced XL, and Maxwell® 16 Bench-top DNA extraction systems.
The DNA extraction performance of three low-throughput extraction systems was evaluated. The instruments and respective chemistries all use a similar extraction methodology that involves binding DNA to a coated magnetic resin in the presence of chaotropic salt, washing of the resin to remove undesirable compounds, and elution of DNA from the particles in a low-salt solution. The AutoMate Express™ (Life Technologies Corporation, Carlsbad, CA), EZ1® Advanced XL (Qiagen Inc., Valencia, CA), and Maxwell® 16 (Promega Corporation, Madison, WI) were compared using a variety of samples including: blood on swabs, blood on denim, blood on cotton, blood mixed with inhibitors (a mixture of indigo, hematin, humic acid, and urban dust) on cotton, blood on FTA® paper, saliva residue on cigarette butt paper, epithelial cells on cotton swabs, neat semen on cotton, hair roots, bones, and teeth. Each instrument had a recommended pre-processing protocol for each sample type, and these protocols were followed strictly to reduce user bias. All extractions were performed in triplicate for each sample type. The three instruments were compared on the basis of quantity of DNA recovered (as determined by real-time PCR), relative level of inhibitors present in the extract (shown as shifts in the C(T) value for the internal PCR control in the real-time PCR assay), STR peak heights, use of consumables not included in the extraction kits, ease of use, and application flexibility. All three systems performed well; however extraction efficiency varied by sample type and with the preprocessing protocol applied to the various samples.


Spatial heterogeneity regulates plant-pollinator networks across multiple landscape scales.
Mutualistic plant-pollinator interactions play a key role in biodiversity conservation and ecosystem functioning. In a community, the combination of these interactions can generate emergent properties, e.g., robustness and resilience to disturbances such as fluctuations in populations and extinctions. Given that these systems are hierarchical and complex, environmental changes must have multiple levels of influence. In addition, changes in habitat quality and in the landscape structure are important threats to plants, pollinators and their interactions. However, despite the importance of these phenomena for the understanding of biological systems, as well as for conservation and management strategies, few studies have empirically evaluated these effects at the network level. Therefore, the objective of this study was to investigate the influence of local conditions and landscape structure at multiple scales on the characteristics of plant-pollinator networks. This study was conducted in agri-natural lands in Chapada Diamantina, Bahia, Brazil. Pollinators were collected in 27 sampling units distributed orthogonally along a gradient of proportion of agriculture and landscape diversity. The Akaike information criterion was used to select models that best fit the metrics for network characteristics, comparing four hypotheses represented by a set of a priori candidate models with specific combinations of the proportion of agriculture, the average shape of the landscape elements, the diversity of the landscape and the structure of local vegetation. The results indicate that a reduction of habitat quality and landscape heterogeneity can cause species loss and decrease of networks nestedness. These structural changes can reduce robustness and resilience of plant-pollinator networks what compromises the reproductive success of plants, the maintenance of biodiversity and the pollination service stability. We also discuss the possible explanations for these relationships and the implications for landscape planning in agricultural areas.


Response of methanogenic archaeal community to nitrate addition in rice field soil.
Addition of nitrate strongly inhibits CH4 production from anoxic soil. The main mechanisms were considered to be the substrate competition and the toxic effect of denitrification intermediates on the methanogenesis. However, it is unclear whether these inhibitory effects are reflected in the structure and dynamics of methanogenic community in the soil. In the present study, the response of methanogenic archaeal community to nitrate addition was determined using terminal restriction fragment length polymorphisms in combination with cloning and sequencing of archaeal 16S rRNA genes. When nitrate was added at the beginning of an anoxic incubation of rice field soil, denitrification occurred rapidly and the denitrification intermediates were detected only for a short time. Total production of CH4 was reduced, but no obvious effect on the structure of methanogenic community was observed. In contrast, when nitrate was added 20 days after the anoxic incubation, the denitrification intermediates obviously accumulated. CH4 production was completely suppressed for 7 and 16 days from treatments of 5 and 10 mM nitrate respectively. The dynamics of methanogenic community also diverged greatly from the control. While the hydrogenotrophic methanogens increased and the acetoclastic methanogens decreased with the incubation in the control soil, the structure and abundance of the methanogenic community remained unchanged after the addition of nitrate. Methanogenesis resumed when the denitrification intermediates were depleted in soil. The analysis of carbon isotopic signals revealed that hydrogenotrophic methanogenesis recovered faster than acetoclastic methanogenesis. Our study suggests that the accumulation of denitrification intermediates has a strong inhibitory effect on the activity but not the structure of methanogenic community.


Effects of aromatherapy massage on blood pressure and lipid profile in korean climacteric women.
This study investigated the effects of aromatherapy massage on the lipid profile and blood pressure in Korean climacteric women. A wait-listed control group, pretest-posttest design was used. The subjects comprised 58 climacteric women: 30 in the experimental group and 28 in the control group. Aromatherapy massage using lavender, rose geranium, rose, and jasmine was given to the experimental group only. Each massage session lasted 30 minutes, and was performed once weekly for two 8-week periods with self abdominal daily massage at home. The intervention produced significant differences in the systolic blood pressure compare to pretreatment and significant differences in systolic and diastolic blood pressures at posttreatment between the two groups. These results suggest that aromatherapy massage may exert positive effects on blood pressure. However, more objective, clinical measures should be applied in a future study with a randomized placebo-controlled design.


Relative weight and race influence average age at menarche: results from two nationally representative surveys of US girls studied 25 years apart.
1) To establish with nationally representative US data whether menarche occurred earlier in the 1990s than it had 25 years before. 2) To assess whether the occurrence of menarche in relation to weight status and race had changed over this time period. Relative weight, race, and menarcheal status of girls in the National Health Examination Survey cycles II and III (1963-1970) were compared with results from the Third National Health and Nutrition Examination Survey (1988-1994). Probit analysis was used to determine the average age at menarche during the 2 survey periods. Logistic regression was used to assess the association of relative weight to likelihood of having reached menarche. The average age at menarche dropped from 12.75 to 12.54 years, and the percentage of girls between 10 and 15 years old who were above the 85th percentile for body mass index increased from 16% to 27% over the 25 years between the 2 surveys. Higher relative weight was strongly associated with increased likelihood of having reached menarche after controlling for age and race. Black girls had a lower average age at menarche than did white girls, which was independent of the effect of relative weight. These analyses from 2 nationally representative samples of US girls suggest a drop of about 2(1/2) months in the average age of menarche during the time period between 1963-1970 and 1988-1994. This was paralleled by a concurrent shift in the population distribution of body mass index z-score toward higher relative weights.


Limited genetic diversity among genotypes of Enterocytozoon bieneusi strains isolated from HIV-infected patients from Sydney, Australia.
Microsporidia are intracellular parasites, with over 1200 species belonging to 143 genera described to date. They are opportunistic pathogens in humans and can cause chronic diarrhoea in immunosuppressed patients. Both Enterocytozoon bieneusi and Encephalitozoon intestinalis cause intestinal disease, with Enterocytozoon bieneusi more commonly identified in patients with human immunodeficiency virus (HIV) infection. In this study, intestinal microsporidial clinical isolates from patients in Sydney, Australia, were genotyped. All specimens were from HIV-infected men with low CD4(+) T-cell counts (<100 cells mm(-3)). Genotyping of the internal transcribed spacer regions of the rRNA gene showed the presence of only one genotype, the anthroponotic Enterocytozoon bieneusi genotype B strain. This study thus highlighted the limited genetic diversity among Australian Enterocytozoon bieneusi isolates, and it is hypothesized that, due to the reduced incidence of microsporidia and the subsequent reduction in the human reservoir of the anthroponotic genotype B, locally acquired intestinal microsporidiosis will rarely be seen in HIV-infected persons undergoing highly active antiretroviral therapy in the future in Australia.


[14-year experience with implantable cardioverter/defibrillators: determination of prognosis and discharge behavior].
The treatment of life threatening ventricular arrhythmias with implantable cardioverter/defibrillators (ICD) has become the therapy of choice; the survival benefit of ICD treatment compared to drug therapy in patients with aborted sudden cardiac death (SCD) and hemodynamically unstable ventricular tachycardia has been proven. In addition for the primary prevention of SCD in high risk patients, ICD therapy is gaining growing acceptance. We analyzed the long-term follow-up of 274 consecutive patients (211 male, 63 female, age 59 +/- 12 years, left ventricular ejection fraction 39 +/- 15%) provided with an ICD between 1984 and 1998. The aim of the study was to ascertain the survival rate in different subgroups and to discover determining factors of ICD discharge and prognosis. Long-term survival probability at 10 resp. 14 years was 84 resp. 65% for the total collective, and the freedom of event probability (neither shocks nor antitachycardiac pacing from the ICD) to 28% each. The risk to die from SCD was below 3% over time. The most pronounced differences regarding prognosis ensued from dividing the collective into heart insufficiency stages. Thus in NYHA class I and II versus III and IV, the cumulative event rate was 61% vs 82% at 5 years, and survival rate amounted to 94 vs 63% at 5 years and 87% vs 30% at 14 years (p < 0.001). Calculating the relative benefit of ICD therapy survival benefit provided by the ICD was shown to decrease significantly after 5 years for patients in NYHA class III/IV, while it increased progressively for patients in NYHA class I/II up to 10 years. Additional determinants of prognosis and ICD discharge rate were identified left ventricular ejection fraction, age and tendency for the basic cardiac disease, however neither the result of electrophysiological testing nor the results of non-invasive risk stratification. In patients with ischemic heart disease, revascularization procedures improved prognosis only in tendency, while the effect of ICD therapy was significant. In patients with the non-obstructive form of hypertrophic cardiomyopathy ICD, discharges occurred in about 50% of patients; in contrast patients with surgical myectomy for obstructive cardiomyopathy showed no events during follow-up. In patients with chronic inflammatory heart disease and normal left ventricular function (LVF), a very low event rate was expected if patients were treated by immunosuppressive drugs. Patients with dilated cardiomyopathy did not differ from patients with ischemic heart disease with respect to prognosis and ICD discharge rate. Significant determinants of prognosis and ICD discharge rate are left ventricular function, age and with limitations the basic cardiac disease. In contrast to patients with better LVF relative survival benefit decreases significantly after 5 years in patients with a worse LVF. Patients with aborted SCD and preserved LVF experience half the ICD discharges compared to patients with poor LVF and gain at the same time a normalization of life expectancy. Causative treatment of the basic disease has an impact on the overall prognosis and event rate, but should in general not influence the decision for IDC implantation in high risk patients.


Understandings of acupuncture application and mechanisms.
Acupuncture involves the stimulation of acupoints, which are located at specific sites of the human body, by insertion of fine metal needles, followed by manipulation. Acupuncture has been proven to be an effective treatment in pain relief. Available evidence showed that acupuncture alleviates acute pain in conditions such as postoperative pain, acute back pain, labour pain, primary dysmenorrhea, tension-type headaches and migraines. In addition, acupuncture relieves chronic pain, for example, low back pain (LBP), knee osteoarthritis (KOA), headache, shoulder pain, and neck pain. For other diseases like insomnia, drug addiction and stroke, more high-quality randomized control trials (RCTs) are needed to confirm the efficacy of acupuncture, although there are particular difficulties surrounding adequate blinding and control group designs. Recent biomedical technology unveils the mechanisms of acupuncture. Studies have found that adenosine triphosphate (ATP) and transient receptor potential vanilloid (TRPV) channels are involved in the stimulation of acupuncture at the acupoint area. In the central nervous system (CNS), neurotransmissions including opioids, serotonin, norepinephrine, orexin and endocannabinoid are modulated by acupuncture to induce analgesia. Moreover, acupuncture reduces cyclooxygenase-2 (COX-2) and prostaglandin E2 (PGE2) levels on the peripheral level by acting on the hypothalamic-pituitary-adrenal (HPA) axis, mediating peripheral opioid release. Acupuncture helps to treat insomnia by inhibiting sympathetic activity and down-regulating the HPA axis. Additionally, acupuncture reduces the effects of positive and negative reinforcements by modulating dopamine release in the nucleus accumbens. Recently, i-needles have been developed to allow for the analysis of metagenomics, meta-transcriptomics, and host-microbiome relationships following acupuncture, while skin implantable microsensors or needle-shaped microsensors are feasible for monitoring real-time microenvironmental changes in acupoints and even target organs. These studies may further accelerate the understanding of acupuncture's action mechanism.


Spin decontamination of broken-symmetry density functional theory calculations: deeper insight and new formulations.
This work re-examines the problem of the broken-symmetry Density-Functional Theory (DFT) solutions in diradical systems, in particular for the calculation of magnetic couplings. The Ms = 0 solution is not an eigenfunction of the S(2) spin operator and the evaluation of the singlet state energy requires a spin-decontamination. A popular approximation is provided by the so-called Yamaguchi formula, which operates using the expectation values of S(2) relative to both Ms = 1 and Ms =0 solutions. Referring to a previous decomposition of the magnetic coupling in terms of direct exchange, kinetic exchange and core polarization, it is shown that this expression will lead to unreliable values of the singlet-triplet energy gap when the spin polarization of the core orbitals becomes large. The here-proposed method of spin-decontamination is based on the Effective Hamiltonian Theory and uses the overlap between the two degenerate Ms = 0 solutions. An approximate and convenient formula, which uses the expectation values of S(2) of the Ms = 0 solutions before and after core polarization is proposed, which is free from the Yamaguchi's formula artefact, as illustrated on an organic diradical presenting a very high value of 〈S(2)〉 for the Ms = 0 solution, the antiferromagnetic coupling being due to the spin polarization mechanism.


Carrier frequency of CYP1B1 mutations in the United States (an American Ophthalmological Society thesis).
CYP1B1 mutations cause autosomal recessive congenital glaucoma. Disease risk assessment for families with CYP1B1 mutations requires knowledge of the population mutation carrier frequency. The purpose of this study is to determine the CYP1B1 mutation carrier frequency in clinically normal individuals residing in the United States. Because CYP1B1 mutations can exhibit variable expressivity, we hypothesize that the mutation carrier frequency is higher than expected. Two hundred fifty individuals without glaucoma or a family history of glaucoma were enrolled. CYP1B1 mutations were identified by DNA sequencing, and pathogenicity was estimated by PolyPhen-2 or a previous report of disease causality. Based on the disease frequency (1 in 10,000) and prevalence of CYP1B1-related congenital glaucoma (15% to 20%), the frequency of CYP1B1-related congenital glaucoma in the United States is approximately 1 in 50,000. Assuming Hardy-Weinberg equilibrium, the expected CYP1B1 mutation carrier frequency would be 1 in 112, or 0.89%. Among the 250 study participants, 11 (4.4%) are carriers of a single pathogenic mutation, representing a carrier frequency of 1 in 22, which is 5.1 times the expected frequency. A higher-than-expected carrier frequency (1 in 33, 3.0%) was also observed in 4300 white individuals sequenced by the National Heart Lung and Blood Institute Exome Sequencing Project. Our results show that the CYP1B1 mutation carrier frequency in the US population is between 1 in 22 and 1 in 33, which is 5.1 to 3.4 times the expected frequency. These results suggest that more individuals than expected are carriers of a deleterious CYP1B1 mutation, and that the prevalence of CYP1B1-related disease may be higher than expected.


Congenital toxoplasmosis. The Toxoplasmosis Study Group.
Congenital toxoplasmosis is a preventable and treatable disease with predominant neurological and ophthalmologic manifestations. These manifestations and approaches to diagnosis, treatment, and prevention of this infection are considered. Outcomes with and without treatment are emphasized. The following also are discussed: recent advances in understanding the organism; its life cycle, epidemiology, pathogenesis, and pathology; and areas in which improvements in diagnosis, treatment, and prevention are needed.


Low-temperature atomic layer deposition delivers more active and stable Pt-based catalysts.
We tailored the size distribution of Pt nanoparticles (NPs) on graphene nanoplatelets at a given metal loading by using low-temperature atomic layer deposition carried out in a fluidized bed reactor operated at atmospheric pressure. The Pt NPs deposited at low temperature (100 °C) after 10 cycles were more active and stable towards the propene oxidation reaction than their high-temperature counterparts. Crucially, the gap in the catalytic performance was retained even after prolonged periods of time (>24 hours) at reaction temperatures as high as 450 °C. After exposure to such harsh conditions the Pt NPs deposited at 100 °C still retained a size distribution that is narrower than the one of the as-synthesized NPs obtained at 250 °C. The difference in performance correlated with the difference in the number of facet sites as estimated after the catalytic test. Our approach provides not only a viable route for the scalable synthesis of stable supported Pt NPs with tailored size distributions but also a tool for studying the structure-function relationship.


Effect of osmotic shock and shearing forces on ferric enterochelin transport in Escherichia coli K12.
The fes mutation in Escherichia coli K12, which inactivates enterochelin esterase, allows the cell to accumulate ferric enterochelin. The ferric complex of enterochelin was released in significant quantities from a fes mutant after osmotic shock. Analysis of the effects of the individual stages of the shock procedure in wild-type cells showed that prior exposure of cells to sucrose and EDTA was not required, careful dilution of cells into a hypo-osmolar medium being sufficient to induce efflux of Fe3+. Prior treatment with EDTA or exposure to shearing forces served either to enhance efflux or to induce efflux in isotonic media. Neither vitamin B12 nor 5'-nucleotidase was released from the periplasm by these procedures. The release observed under mild conditions was stimulated specifically by Co2+, did not occur at 0 degree C, and was inhibited by 2,4-dinitrophenol at 37 degrees C. From these observations, it was concluded that the efflux of Fe3+ represents a physiological response of the cell to exposure to a hypo-osmolar medium. Such changes may enhance survival following physicochemical stressing of the bacterial outer membrane.


Synthesis and structural determination of zinc complexes based on an anilido-aldimine ligand containing an O-donor pendant arm: zinc alkoxide derivative as an efficient initiator for ring-opening polymerization of cyclic esters.
Zinc complexes bearing the anilido-aldiminate AA(OMe) ligand (AA(OMe)-H = (E)-2,6-diisopropyl-N-(2-(((2-methoxyethyl)imino)methyl)phenyl)aniline) were synthesized in a stepwise method and were structurally characterized. The reaction of AA(OMe)-H (1) with one equivalent of diethyl zinc (ZnEt2) furnishes a three-coordinated and mononuclear zinc complex [(AA(OMe))ZnEt] (2). Further reaction of 2 with a stoichiometric amount of benzyl alcohol (BnOH) affords a four-coordinated and dinuclear zinc benzylalkoxide complex [(AA(OMe))Zn(μ-OBn)]2 (3). In the presence of two equivalents of AA(OMe)-H with ZnEt2, a homoleptic and four-coordinated zinc complex [(AA(OMe))2Zn] (4) is formed. The geometry around the zinc centres of 3 and 4 are both distorted tetrahedrals, while 2 adopts a different coordination mode with a slightly distorted trigonal planar geometry. The variable-temperature (1)H NMR studies of 3 illustrate that 3 exhibits a dinuclear structure in solution at low temperature as well as in the solid state. While raising the temperature, it drifts towards dissociation to form a mononuclear zinc benzylalkoxide species, which coexists in solution. The ring-opening polymerizations of ε-caprolactone (ε-CL) and β-butyrolactone (β-BL) catalyzed by complexes 3 and 4 are investigated. The ε-CL and β-BL polymerizations initiated by zinc alkoxide 3 were demonstrated to have living characteristics and to proceed in a controlled manner with narrow polydispersity indices (PDIs < 1.12). An efficient catalytic performance for the β-BL polymerization with a high monomer-to-initiator ratio (1200/1) initiated by 3 has also been reported.


Current siRNA targets in atherosclerosis and aortic aneurysm.
Atherosclerosis (ATH) and aortic aneurysms (AA) remain challenging chronic diseases that confer high morbidity and mortality despite advances in medical, interventional, and surgical care. RNA interference represents a promising technology that may be utilized to silence genes contributing to ATH and AA. Despite positive results in preclinical and some clinical feasibility studies, challenges such as target/sequence validation, tissue specificity, transfection efficiency, and mitigation of unwanted off-target effects remain to be addressed. In this review the most current targets and some novel approaches in siRNA delivery are being discussed. Due to the plethora of investigated targets, only studies published between 2010 and 2014 were included.


Analysis of the virulence, infection process, and extracellular enzyme activities of Aspergillus nomius against the Asian corn borer, Ostrinia furnacalis guenée (Lepidoptera: Crambidae).
The control of Ostrinia furnacalis, a major pest of maize in Xinjiang, is challenging owing to the occurrence of resistant individuals. Entomopathogenic fungi (EPF) are natural insect regulators used as substitutes for synthetic chemical insecticides. The fungus Aspergillus nomius is highly pathogenic to O. furnacalis; however, its virulence characteristics have not been identified. This study aimed to analyse the lethal efficacy, mode of infection on the cuticle, and extracellular enzyme activity of A. nomius against O. furnacalis. We found that the mortality and mycosis of O. furnacalis were dose-dependent when exposed to A. nomius and varied at different life stages. The egg-hatching and adult emergence rates decreased with an increase in conidial suspension. The highest mortality (83.33%, 7 d post-infection [DPI]) and mycosis (74.33%, 7 DPI) and the lowest mortality response (8.52 × 103 conidia mL-1) and median lethal time (4.91 d) occurred in the 3rd instar larvae of O. furnacalis. Scanning electron microscopy indicated that numerous conidia germination and infection structure formation may have contributed to the high pathogenicity of A. nomius against O. furnacalis. There were significant correlations between O. furnacalis mortality and the activities of extracellular protease, lipase, and chitinase of A. nomius. This study revealed the infection process of the highly pathogenic A. nomius against O. furnacalis, providing a theoretical basis and reference for strain improvement and field application of EPF.


Mutations in EXPH5 result in autosomal recessive inherited skin fragility.
Several different genes have been implicated in the pathophysiology of inherited blistering skin diseases. Recently, autosomal recessive loss-of-function mutations in EXPH5 (encoding exophilin-5, also known as Slac2-b, a protein involved in intracellular vesicle transport) were identified in a new mechanobullous disease resembling a form of epidermolysis bullosa simplex (EBS). Here, we searched for mutations in EXPH5 in a 4-year-old white boy with EBS in whom initial Sanger sequencing of known genes implicated in intraepidermal skin fragility failed to identify pathogenic mutations. Transmission electron microscopy of rubbed nonlesional patient skin revealed disruption of keratinocytes in the lower epidermis with cytolysis and acantholysis, keratin filament clumping and prominent perinuclear cytoplasmic vesicles, and provided the clue to the candidate gene pathology. Sanger sequencing of genomic DNA showed compound heterozygosity for two new mutations in EXPH5, c.1947dupC (p.Pro649fsPro*11) and c.2249C>A (p.Ser750*). Immunofluorescence microscopy of patient skin showed a complete absence of exophilin-5 labelling. This case represents the third pedigree with EXPH5 mutations resulting in inherited skin fragility. The clinical and molecular data expand genotype-phenotype correlation in this new form of EBS and demonstrate the important role of exophilin-5 in keratinocyte cell biology.


Staging laparoscopy for pancreatic cancer should be used to select the best means of palliation and not only to maximize the resectability rate.
Staging laparoscopy, based on the assumption that endobiliary stenting is the best palliation, allegedly saves an "unnecessary" laparotomy for incurable pancreatic cancer. Our aim was to determine survival of patients with clinically resectable pancreatic cancer that is found to be unresectable intraoperatively and thereby infer appropriate utilization of staging laparoscopy. A retrospective analysis was undertaken of 148 patients with ductal adenocarcinoma (1985 to 1992) with a clinically resectable lesion based on current imaging techniques. All were considered candidates for resection but were deemed unresectable at operation because of metastases to the liver (group I; 29 patients), the peritoneum (group II; 22 patients), or distant lymph nodes (group III; 44 patients) or because of vascular invasion (group IV; 53 patients). Overall median survival was 9 months (range 1 to 53 months), but by group was as follows: group I, 6 months; group II, 7 months; group III, 11 months; and group IV, 11 months. Individual comparisons showed shorter survival for patients with distant nodal, liver, or peritoneal metastases than with nodal or vascular involvement (P<0.03). Staging laparoscopy should be performed to identify patients with liver or peritoneal metastases who have an expected survival of approximately 6 months, in whom short-term endoscopic palliation is satisfactory. Extended laparoscopy to identify lymph node or vascular involvement is contingent upon which palliation (operative vs. endoscopic) is considered most appropriate. Because we believe operative bypass provides better, more durable palliation in this latter group, we have not adopted extended laparoscopy.


25-Hydroxyvitamin D3 1-α-hydroxylase-dependent stimulation of renal klotho expression by spironolactone.
Klotho, a transmembrane protein, protease and hormone mainly expressed in kidney, is required for the suppression of 1,25(OH)2D3-generating 25-hydroxyvitamin D3 1-alpha-hydroxylase (Cyp27b1) by FGF23. Conversely, 1,25(OH)2D3 stimulates, by activating the vitamin D3 receptor (Vdr), the expression of klotho, thus establishing a negative feedback loop. Klotho protects against renal and vascular injury. Klotho deficiency accelerates aging and early death, effects at least partially due to excessive formation of 1,25(OH)2D3 and subsequent hyperphosphatemia. Klotho expression is inhibited by aldosterone. The present study explored the interaction of aldosterone and DOCA as well as the moderately selective mineralocorticoid receptor antagonist spironolactone on klotho expression. mRNA levels were determined utilizing quantitative RT-PCR in human embryonic kidney cells (HEK293) or in renal tissues from mice without or with prior mineralocorticoid (aldosterone or DOCA) and/or spironolactone treatment. In HEK293 cells, protein levels were determined by western blotting. The experiments in HEK293 cells were performed without or with silencing of CYP27B1, of vitamin D3 receptor (VDR) or of mineralocorticoid receptor (NR3C2). In HEK293 cells aldosterone and in mice DOCA significantly decreased KLOTHO gene expression, effects opposed by spironolactone treatment. Spironolactone treatment alone significantly increased KLOTHO and CYP27B1 transcript levels in HEK293 cells (24 hours) and mice (8 hours or 5 days). Moreover, spironolactone significantly increased klotho and CYP27B1 protein levels in HEK293 cells (48 hours). Reduced NR3C2 expression following silencing did not significantly affect KLOTHO and CYP27B1 transcript levels in presence or absence of spironolactone. Silencing of CYP27B1 and VDR significantly blunted the stimulating effect of spironolactone on KLOTHO mRNA levels in HEK293 cells. Besides blocking the effects of aldosterone, spironolactone upregulates KLOTHO gene expression by upregulation of 25-hydroxyvitamin D3 1-alpha-hydroxylase with subsequent activation of the vitamin D3 receptor by 1,25(OH)2D3, an effect possibly independent from the mineralocorticoid receptor.


A common telomeric gene silencing assay is affected by nucleotide metabolism.
Telomere-associated position-effect variegation (TPEV) in budding yeast has been used as a model for understanding epigenetic inheritance and gene silencing. A widely used assay to identify mutants with improper TPEV employs the URA3 gene at the telomere of chromosome VII-L that can be counterselected with 5-fluoroorotic acid (5-FOA). 5-FOA resistance has been inferred to represent lack of transcription of URA3 and therefore to represent heterochromatin-induced gene silencing. For two genes implicated in telomere silencing, POL30 and DOT1, we show that the URA3 telomere reporter assay does not reflect their role in heterochromatin formation. Rather, an imbalance in ribonucleotide reductase (RNR), which is induced by 5-FOA, and the specific promoter of URA3 fused to ADH4 at telomere VII-L are jointly responsible for the variegated phenotype. We conclude that metabolic changes caused by the drug employed and certain mutants being studied are incompatible with the use of certain prototrophic markers for TPEV.


Quantitative microautoradiography of arteries: comparison of radioactivity to silver.
The local concentration of silver in developed aortic transmural microautoradiographs was compared to the corresponding 125I-labeled albumin radioactivity concentration [ci(x)] in the subjacent tissue. Silver [s(x)] was measured by electron probe X-ray microanalysis (EPA) and the corresponding ci(x) by direct gamma-ray counting. The results show 1) that the relationship between volume-averaged values of radioactivity (ci) and EPA signal (s) is adequately described by ci = Ks, where K is a proportionality constant, and 2) that ci(x) measured by EPA [i.e., Ks(x)] agrees closely with ci(s) measured direcly from en face microtomy slices of corresponding unfixed tissue specimens.


Clonidine Maintenance Prolongs Opioid Abstinence and Decouples Stress From Craving in Daily Life: A Randomized Controlled Trial With Ecological Momentary Assessment.
The authors tested whether clonidine blocks stress-induced seeking of heroin and cocaine. The study was also intended to confirm translational findings from a rat model of drug relapse by using ecological momentary assessment of patients' stress to test hypotheses about clonidine's behavioral mechanism of action. The authors conducted a randomized double-blind placebo-controlled clinical trial with 208 opioid-dependent patients at an outpatient buprenorphine clinic. The 118 participants (57%) who maintained abstinence during weeks 5-6 were continued on buprenorphine and randomly assigned to receive clonidine (N=61) or placebo (N=57) for 14 weeks. Urine was tested thrice weekly. Lapse was defined as any opioid-positive or missed urine test, and relapse as two or more consecutive lapses. Time to lapse and relapse were examined with Cox regressions; longest period of abstinence was examined with a t test, and ecological momentary assessment data were examined with generalized linear mixed models. In an intent-to-treat analysis, clonidine produced the longest duration (in consecutive days) of abstinence from opioids during the intervention phase (34.8 days [SD=3.7] compared with 25.5 days [SD=2.7]; Cohen's d=0.38). There was no group difference in time to relapse, but the clonidine group took longer to lapse (hazard ratio=0.67, 95% CI=0.45-1.00). Ecological momentary assessment showed that daily-life stress was partly decoupled from opioid craving in the clonidine group, supporting the authors' hypothesized mechanism for clonidine's benefits. Clonidine, a readily available medication, is useful in opioid dependence not just for reduction of withdrawal signs, but also as an adjunctive maintenance treatment that increases duration of abstinence. Even in the absence of physical withdrawal, it decouples stress from craving in everyday life.


Orphan nuclear receptor TLX regulates astrogenesis by modulating BMP signaling.
Neural stem cells (NSCs) are self-renewing multipotent progenitors that generate both neurons and glia. The precise control of NSC behavior is fundamental to the architecture and function of the central nervous system. We previously demonstrated that the orphan nuclear receptor TLX is required for postnatal NSC activation and neurogenesis in the neurogenic niche. Here, we show that TLX modulates bone morphogenetic protein (BMP)-SMAD signaling to control the timing of postnatal astrogenesis. Genes involved in the BMP signaling pathway, such as Bmp4, Hes1, and Id3, are upregulated in postnatal brains lacking Tlx. Chromatin immunoprecipitation and electrophoretic mobility shift assays reveal that TLX can directly bind the enhancer region of Bmp4. In accordance with elevated BMP signaling, the downstream effectors SMAD1/5/8 are activated by phosphorylation in Tlx mutant mice. Consequently, Tlx mutant brains exhibit an early appearance and increased number of astrocytes with marker expression of glial fibrillary acidic protein (GFAP) and S100B. Taken together, these results suggest that TLX tightly controls postnatal astrogenesis through the modulation of BMP-SMAD signaling pathway activity.


A genome wide search for susceptibility loci in three European malignant hyperthermia pedigrees.
Malignant hyperthermia (MH) is an autosomal dominant disorder which is potentially lethal in susceptible individuals on exposure to commonly used inhalational anaesthetics and depolarising muscle relaxants. Crises reflect the consequences of disturbed skeletal muscle calcium homeostasis. Susceptibility was first localised to chromosome 19q13.1 and the skeletal muscle ryanodine receptor, RYR1 (the calcium release channel of the sarcoplasmic reticulum). Defects in this gene have been identified which cosegregate with the MHS phenotype and evidence as to their potential causal roles has accumulated. MH has, however, been shown to be genetically heterogeneous, additional loci on chromosomes 3q, 17q and 7q being proposed. Pedigrees remain in Europe where linkage status is still unclear. In a collaborative search of the human genome conducted with three pedigrees whose disease status was classified according to the European IVCT protocol we have evidence to suggest that at least two further loci exist for MH susceptibility. One of these locates to chromosome 1q, the site of a candidate gene, CACNL1A3, encoding the alpha-subunit of the dihydropyridine receptor. The second region resides on chromosome 5p to where no known candidate has been mapped to date. The third family exhibited inconclusive results which suggests the existence of at least one other locus. This study adds to the evidence for considerable genetic heterogeneity in MH and will provide a route to further our understanding of the molecular pathology of the condition.


[Femur fractures caused by long-term bisphosphonate therapy].
There is increasingly more evidence to suggest that spontaneous femur fractures may be an adverse effect of long-term bisphosphonate use. Patients typically present with prodromal thigh pain, cortical thickening on conventional radiographs or present at A&E with a spontaneous transverse subtrochanteric femur fracture with a medial spike. Discontinuation of bisphosphonate therapy is advisable in patients presenting with a spontaneous femur fracture with specific features while using bisphosphonates. The way in which bisphosphonates alter the mechanical properties of bone is as yet unknown.


Risk factor synergism in aneurysmal subarachnoid hemorrhage: a cross-sectional study.
Spontaneous subarachnoid hemorrhage (SAH) accounts for 5-10% of strokes but a disproportionately large amount of stroke-related morbidity. Several risk factors have been described, including smoking, hypertension, increasing age, and female sex. This cross-sectional study examined all patients with aSAH within a nationally representative catchment from 01/01/2017 to 31/12/2020. Patients with aneurysmal SAH were identified from multiple sources, including a prospective database and death records. The population was estimated from projections from a door-to-door census and risk factors from stratified random sampled surveys conducted on a yearly basis. Poisson regression models were used to estimate the incidence and incidence rate ratios (IRRs) for risk factors with 95% confidence intervals (95% CIs). We identified 875 cases of aSAH in 11,666,807 patient-years of follow-up, which corresponded to a crude incidence of 7.5 per 100,000 patient-years (95% CI 7-8) and a standardized incidence of 6.1/100,000 (95% CI 5.6-6.5). Smoking was the strongest individual risk factor, with a standardized incidence of 24/100,000 (95% CI 20-27) in smokers compared with 2.6/100,000 (2.1-3.2) in non-smokers (age-adjusted IRR 9.2, 95% CI 6.3-13.6). Hypertension (age-adjusted IRR 3.1, 95% CI 2.2-4.3) and female sex (age-adjusted IRR 1.8, 95% CI 1.4-2.3) were also associated with increased incidence. The highest incidence was observed in hypertensive smokers (standardized incidence 63/100,000, 95% CI 41-84), who had a lifetime risk of aSAH of 6.7% (95% CI 5.4-8.1) after age 35. Compared with participants who were non-smokers without hypertension, the age-adjusted IRR in hypertensive smokers was 27.9 (95% CI 15.9-48.8). Smoking is the most prominent individual risk factor for aSAH. Smoking and hypertension appear to interact to increase the risk of aSAH synergistically.


Diabetic patients are almost twice as likely to experience complications from arthroscopic rotator cuff repair.
Large population-based studies have demonstrated increased prevalence of rotator cuff disease amongst diabetics. Recent studies have suggested comparable clinical outcomes from rotator cuff repair despite suggestions of increased complication rates amongst diabetics. However, there is a relative paucity of studies considering the effect of diabetes upon complication rate. We aim to report and quantify the effect of diabetes on complication rates following arthroscopic rotator cuff repair. A retrospective review of a consecutive series of patients undergoing arthroscopic rotator cuff repair between January 2011 and December 2014 was performed. Diabetic status and complication data defined as infection, frozen shoulder, re-tear or re-operation were collected and interrogated. A total of 462 patients were included at median follow-up of 5.6 years. Diabetics were significantly more likely to experience frozen shoulder (15.8% vs. 4.4%, p = 0.001), re-tear (26.3% vs. 15.6%, p = 0.042) or at least one complication following surgery (35.1% vs. 22.7%, p = 0.041) compared to non-diabetics. These equated to odds ratios of 4.03, 1.94 and 1.84, respectively. Diabetic patients are almost twice as likely to experience complications following arthroscopic rotator cuff repair, including double the risk of repair failure and more than four times the risk of frozen shoulder.


Endoscopic endonasal transplanum approach to the paraclinoid internal carotid artery.
Objective To investigate the relevance of an endoscopic transnasal approach to the surgical treatment of paraophthalmic aneurysms. Setting Binasal endoscopic transplanum surgery was performed. Participants Seven cadaver heads were studied. Main Outcome Measures (1) Dimensions of the endonasal corridor, including the operative field depth, lateral limits, and the transplanum craniotomy. (2) The degree of vascular exposure. (3) Surgical maneuverability and access for clip placements. Results The mean operative depth was 90 ± 4 mm. The lateral corridors were limited proximally by the alar rim openings (29 ± 4 mm) and distally by the distance between the opticocarotid recesses (19 ± 2 mm). The mean posteroanterior distance and width of the transplanum craniotomy were 19 ± 2 mm and 17 ± 3 mm, respectively. Vascular exposure was achieved in 100% of cases for the clinoidal internal carotid artery (ICA), ophthalmic artery, superior hypophyseal artery, and the proximal ophthalmic ICA. Surgical access and clip placement was achieved in 97.6% of cases for vessels located anterior to the pituitary stalk (odds ratio [OR] 73.8; 95% confidence interval [CI] 7.66 to 710.8; p = 0.00). Conclusion The endoscopic transnasal approach provides excellent visualization of the paraclinoid region vasculature and offers potential surgical alternative for paraclinoid aneurysms.


Results of one-year antismoking campaign conducted within the framework of multifactorial prevention of ischaemic heart disease.
Among 593 smokers examined within the framework of the programme of multifactorial prevention of ischaemic heart disease [IHD], various smoking control measures were taken for combating smoking (lecture with discussion, group and individual sessions). Smokers with arterial hypertension and/or IHD were given antismoking advice by their physicians. After one year 20.7% persons of the whole group stopped smoking, and 25.3% reduced the number of daily consumed cigarettes by at least one quarter. The total number of smokers decreased by 8.1% (p = 0.0005). In the control group, the smoking habit changed only insignificantly. The greatest success (50% of persons who gave up smoking) was obtained at group discussions, using positive motivation to stop smoking; the smallest effect (12.2%) was reached among patients where the antismoking campaign was entrusted to their physicians. Persons who were given greatest attention in the antismoking campaign (patients participating in group discussions) showed the lowest percentage of those who reported for annual examination within the framework of the programme of multifactorial prevention of IHD.


Duplex in situ hybridization in the study of gene co-regulation in the vertebrate brain.
We describe here a high-sensitivity in situ hybridization protocol, optimized for fresh-frozen brain sections, that enables the detection of two transcripts, at single cell resolution. Riboprobes directed against two mRNAs of interest are synthesized with nucleotides tagged with different haptens (digoxigenin- or biotin-UTP), via in vitro transcription, hybridized simultaneously to brain sections, and independently detected through immunocytochemistry. Sequential detection of each probe involves peroxidase-mediated precipitation of tyramide-linked fluorophores of separate emission wavelengths. In addition, we demonstrate how classic non-fluorescent chromogens, such as 3,3'-diaminobenzidine, can be successfully combined with fluorescence-based detection, to yield reliable detection of two transcript populations. We provide examples of representative results obtained with this protocol and describe necessary controls. Additionally, we discuss common problems associated with this methodology, and detail troubleshooting recommendations. Although this method has been optimized for brain sections, it may be useful to detect two mRNA species in a variety of tissues.


The origin of bmp16, a novel Bmp2/4 relative, retained in teleost fish genomes.
Whole genome sequences have allowed us to have an overview of the evolution of gene repertoires. The target of the present study, the TGFbeta superfamily, contains many genes involved in vertebrate development, and provides an ideal system to explore the relationships between evolution of gene repertoires and that of developmental programs. As a result of a bioinformatic survey of sequenced vertebrate genomes, we identified an uncharacterized member of the TGFbeta superfamily, designated bmp16, which is confined to teleost fish species. Our molecular phylogenetic study revealed a high affinity of bmp16 to the Bmp2/4 subfamily. Importantly, further analyses based on the maximum-likelihood method unambiguously ruled out the possibility that this teleost-specific gene is a product of teleost-specific genome duplication. This suggests that the absence of a bmp16 ortholog in tetrapods is due to a secondary loss. In situ hybridization showed embryonic expression of the zebrafish bmp16 in the developing swim bladder, heart, tail bud, and ectoderm of pectoral and median fin folds in pharyngula stages, as well as gut-associated expression in 5-day embryos. Comparisons of expression patterns revealed (1) the redundancy of bmp16 expression with its homologs in presumably plesiomorphic expression domains, such as the fin fold, heart, and tail bud, which might have permitted its loss in the tetrapod lineage, and (2) the loss of craniofacial expression and gain of swim bladder expression of bmp16 after the gene duplication between Bmp2, -4 and -16. Our findings highlight the importance of documenting secondary changes of gene repertoires and expression patterns in other gene families.


[Effectiveness of super-fractionated irradiation in the radiotherapy of inoperable lung cancer].
The paper is concerned with the results of gamma-beam therapy of 120 patients with inoperable lung cancer using the fractionation of a common daily dose of 2-2.2 Gy into 2 equal fractions as a variant of super-fractionated irradiation used in clinical practice. The direct and short-term therapeutic results were improved: a complete tumor regression rate determined by x-ray was 26.7%, a 2-year survival rate 19.2%. In the group of patients who were irradiated by the commonly used methods, the same values were lower. Improved tolerance of therapy with fractionation of a common daily dose was noted.


Abnormal immune responses in the ocular presentation of Wiskott-Aldrich syndrome.
Wiskott-Aldrich syndrome is an x-linked disorder characterized by a triad of thrombocytopenia, eczema, and recurrent infections. Until recently, survival into the second decade was rare. In the last few years, a better understanding of the syndrome has been achieved, and earlier recognition and newer modes of treatment have improved life. These patients are living longer and ophthalmologic presentation reflects this change. We present three case histories of patients followed for the past 15 years who have had, in association with this syndrome, eczema of the eyelids, episcleritis, marginal keratitis, and blepharitis.


[Interaction of poly A with phospholipid membranes using IR-spectroscopy].
IR-spectra of poly-A phosphatidylcholine complexes in D2O were studied. It has been shown that in the absence of Mg2+ the spectrum was a superposition of spectra of poly-A and phosphatidylcholine. Introduction of MgCl2 to the concentration of 0.03 M led to the dramatic decrease of absorbance at 1629 cm-1 characteristic of backbone vibration of adenine. Further increase of the concentration of MgCl2 up to 0.08 M resulted in the additional absorbance maximum at 1627 cm-1. Under a variety of temperatures the position of the maximum did not change. It was assumed that on adding magnesium ions to the "poly-A-lipid" mixture in D2O the components formed a stable complex. At high Mg2+ concentration a partial disarrangement of stacking interactions in poly-A occurred that might be a consequence of the incorporation of nucleic bases into hydrophobic membrane phase.


[Chronic cough in pediatrics: Review and analysis].
To review and to analyze the existing literature regarding chronic cough in children. We searched published articles in PubMed using the key words: cough, chronic cough, chronic cough in children, persistent cough, gastroesophageal reflux in children. Cough in children is common; the majority of cases reflect respiratory infections. Cough is rarely associated to a serious disorder. All children with cough persisting for more than 8 weeks should be evaluated. A careful history, physical examination, chest x-ray, and spirometry (in an able child) are recommended for all children with chronic cough. If a diagnosis is no evident (nonspecific cough), then an approach based on characterizing the cough as "wet" or "dry" is helpful. In each case, specific etiology-based treatment is recommended when possible; otherwise, a therapeutic trial is indicated with inhaled corticosteroids (ICS) for children with dry cough (for possible asthma) and antibiotics for wet cough (for possible protracted bacterial bronchitis [PPB] or sinusitis). The treatment should be reviewed within the specified time frame for normal response. If no effect is obvious, the treatment should be stopped and alternative diagnosis considered. Multiple etiologies may need to be treated concomitantly. Every effort should be made to identify the underlying cause. Data suggests potential harmful effects of medications used for symptomatic relief of acute or chronic cough in children. Medical history and physical examination, together with appropriate investigations allow the correct diagnosis of chronic cough. The diagnostic approach is summarized in Figures 1 and 2. Most cases can be managed with this algorithm. For the difficult, complex and rare cases referral to a Pediatric specialist in Allergy, Clinical Immunology and Respiratory Diseases is warranted.


Control of Citrus Huanglongbing via Trunk Injection of Plant Defense Activators and Antibiotics.
Citrus huanglongbing (HLB) or greening is a devastating disease of citrus worldwide and no effective control measure is currently available. Plant defense activators environmentally friendly compounds capable of inducing resistance against many plant pathogens. Earlier studies showed that foliar spray of plant defense inducers could slow down HLB disease progress. In this study, eight plant defense activators and three antibiotics were evaluated in three field trials for their effect to control HLB by trunk injection of young and mature sweet orange trees. Results showed that four trunk injections of several activators, including salicylic acid, oxalic acid, acibenzolar-S-methyl, and potassium phosphate, provided significant control of HLB by suppressing 'Candidatus Liberibacter asiaticus' titer and disease progress. Trunk injection of penicillin, streptomycin, and oxytetracycline hydrochloride resulted in excellent control of HLB. In general, antibiotics were more effective in reduction of 'Ca. L. asiaticus' titer and HLB symptom expressions than plant defense activators. These treatments also resulted in increased yield and better fruit quality. Injection of both salicylic acid and acibenzolar-S-methyl led to significant induction of pathogenesis-related (PR) genes PR-1 and PR-2 genes. Meanwhile, injection of either potassium phosphate or oxalic acid resulted in significant induction of PR-2 or PR-15 gene expression, respectively. These results suggested that HLB diseased trees remained inducible for systemic acquired resistance under field conditions. In summary, this study presents information regarding controlling HLB via trunk injection of plant defense activators and antibiotics, which helps citrus growers in decision making regarding developing an effective HLB management program.


Antiapoptotic effect of DDB against hepatic ischemia-reperfusion injury.
Dimethyl-Diphenyl-Bicarboxylate (DDB) is a hepatoprotectant used in treatment of chronic viral hepatitis. The mechanism of DDB against liver injury needs further elucidation. Our objective is to explore whether DDB exerts hepatoprotective effect against hepatic ischemia reperfusion injury (I/R) and to identify its potential effect on apoptotic cell death targeting the extrinsic apoptotic pathway. Rats were divided into four groups: Group I (sham-operated), Group II (I/R group), Group III (DDB Pretreated group): where rats were treated with DDB at 3 dose levels (100 mg/kg, 300 mg/kg, 500 mg/kg) for 10 days, then subjected to I/R and Group IV (DDB treated group): rats were treated with DDB at the same dose levels but without being subjected to I/R. Serum alanine aminotransferase (ALT) & aspartate aminotransferase (AST), tissue levels of GSH, MDA & tumor necrosis factor alpha (TNF-α), activity of signaling Caspases 3, 8, 9 as well as expression of nuclear factor-κB (NF-κB) were measured. End point of apoptosis was detected by DNA fragmentation. DDB administration before I/R, significantly attenuated elevated levels of ALT, AST, MDA and TNF-α and increased GSH content. There were significant decrease in caspases activities, expression of NF-κB as well as DNA fragmentation. DDB administration to normal rats, inhibited lipid peroxidation, enhanced antioxidant activity, decreased TNF-α and downregulated expression of NF-κB. These effects are dose-dependent. DDB exerts potent antioxidant effect in normal and injured livers. It inhibited hepatic I/R induced apoptosis through inhibiting signaling Caspases 3, 8, 9, TNF-α production which regulates the extrinsic apoptotic pathway and down regulates NF-κB expression in liver tissue.


Impact of confining 3-D polymer networks on dynamics of bacterial ingress and self-organisation.
Studies of microbial interactions during motility, micro-structuring and colonisation have predominately been limited to surface associated bacteria involving materials such as semi-solid biomolecular hydrogels and thin liquid films. Recently, these surfaces have been extended to synthetic polymers where they provide defined chemistries and structural properties. However, precise details of microbial ingress into the confined fluid volume of synthetic 3-D hydrogel networks and their subsequent microstructuring remain to be defined. Here, we show that Gram-positive and Gram-negative bacteria internally populate mesoporous polyacrylate hydrogels by quantifying: the dynamic advancing population front and the resultant spontaneous self-organisation into well-defined clusters and micro-colonies. Polymer chain conjugated fluorescent nanoparticles indicated that both bacterial clusters and micro-colonies associated directly with the polymer chains of the mesoporous hydrogel. Protonation of the K-polyacrylate made chains more hydrophobic and globular in conformation, reducing the swelling of the hydrogel by half. However, the bacterial population increased by 30% indicating the dominance of hydrophobic and viscoelastic interactions as well as the cation chemistry within the confined fluids of synthetic polymer hydrogels despite pore size reductions of 50%. Synthetic polymer hydrogels having a range of porosities when swollen together with controllable chemical and structural functionality can potentially offer well-defined microenvironments for bacterial populations in advancing biotechnologies such as inoculants and substrates in the production of therapeutic agents.


Photo-modulation of supramolecular polymorphism in the self-assembly of a scissor-shaped azobenzene dyad into nanotoroids and fibers.
Recent advances in the research field of supramolecularly engineered dye aggregates have enabled the design of simple one-dimensional stacks such as fibers and of closed structures such as nanotoroids (nanorings). More complex and advanced supramolecular systems could potentially be designed using a molecule that is able to provide either of these distinct nanostructures under different conditions. In this study, we introduced bulky but strongly aggregating cholesterol units to a scissor-shaped azobenzene dyad framework, which affords either nanotoroids, nanotubes, or 1D fibers, depending on the substituents. This new dyad with two trans-azobenzene arms shows supramolecular polymorphism in its temperature-controlled self-assembly, leading to not only oligomeric nanotoroids as kinetic products, but also to one-dimensional fibers as thermodynamic products. This supramolecular polymorphism can also be achieved via photo-triggered self-assembly, i.e., irradiation of a monomeric solution of the dyad with two cis-azobenzene arms using strong visible light leads to the preferential formation of nanotoroids, whereas irradiation with weak visible light leads to the predominant formation of 1D fibers. This is the first example of a successful light-induced modulation of supramolecular polymorphism to produce distinctly nanostructured aggregates under isothermal conditions.


Granulosa cell tumours of the ovary.
Granulosa cell tumours are rare, potentially malignant sex cord stromal tumours of the ovary. They are unique in their presentation and histological features. Many of them are hormone-producing and this property helps them to present early unlike other epithelial ovarian cancers. As a result, most of them will be in an early stage at the time of initial diagnosis. The tumour can manifest in young girls as a juvenile form and conservative management with unilateral salpingo-opherectomy may be an option in them as 95% are unilateral. Surgery is the treatment of choice and initial staging laparatomy a determinant recurrence. Advance stage of the tumour, its size (>5 cm), mitotic figures (>10/hpf), nuclear atypia and absence of call-exner bodies are poor prognostic factors. Such tumours are characterised by late recurrences and this necessitates a prolonged follow-up. Tumour markers such as inhibin and estradiol are useful in follow-up. Chemotherapy, radiotherapy and hormone replacement therapy have very little role in the initial treatment and may be suggested in case of recurrences. With appropriate treatment, a better survival rate can be achieved as against other ovarian malignancies. Methods used for locating, selecting and synthesising data: A search of Medline and Cochrane data base for the period from 1999 to 2010 was carried out to include relevant systematic reviews, meta-analysis, randomised controlled and other clinical and rare case reports. The date of the last search was January 2010.


Mutant PIK3CA is a targetable driver alteration in histiocytic neoplasms.
Langerhans cell histiocytosis (LCH) is an inflammatory myeloid neoplasm characterized by the accumulation of clonal mononuclear phagocyte system cells expressing CD1a and CD207. In the past decade, molecular profiling of LCH as well as other histiocytic neoplasms demonstrated that these diseases are driven by MAPK activating alterations, with somatic BRAFV600E mutations in >50% of patients with LCH, and clinical inhibition of MAPK signaling has demonstrated remarkable clinical efficacy. At the same time, activating alterations in kinase-encoding genes, such as PIK3CA, ALK, RET, and CSF1R, which can activate mitogenic pathways independent from the MAPK pathway, have been reported in a subset of histiocytic neoplasms with anecdotal evidence of successful targeted treatment of histiocytoses harboring driver alterations in RET, ALK, and CSF1R. However, evidence supporting the biological consequences of expression of PIK3CA mutations in hematopoietic cells has been lacking, and whether targeted inhibition of PI3K is clinically efficacious in histiocytic neoplasms is unknown. Here, we provide evidence that activating mutations in PIK3CA can drive histiocytic neoplasms in vivo using a conditional knockin mouse expressing mutant PIK3CAH1047R in monocyte/dendritic cell progenitors. In parallel, we demonstrate successful treatment of PIK3CA-mutated, multisystemic LCH using alpelisib, an inhibitor of the alpha catalytic subunit of PI3K. Alpelisib demonstrated a tolerable safety profile at a dose of 750 mg per week and clinical and metabolic complete remission in a patient with PIK3CA-mutated LCH. These data demonstrate PIK3CA as a targetable noncanonical driver of LCH and underscore the importance of mutational analysis-based personalized treatment in histiocytic neoplasms.


Subcortical brain structure and suicidal behaviour in major depressive disorder: a meta-analysis from the ENIGMA-MDD working group.
The aetiology of suicidal behaviour is complex, and knowledge about its neurobiological mechanisms is limited. Neuroimaging methods provide a noninvasive approach to explore the neural correlates of suicide vulnerability in vivo. The ENIGMA-MDD Working Group is an international collaboration evaluating neuroimaging and clinical data from thousands of individuals collected by research groups from around the world. Here we present analyses in a subset sample (n=3097) for whom suicidality data were available. Prevalence of suicidal symptoms among major depressive disorder (MDD) cases ranged between 29 and 69% across cohorts. We compared mean subcortical grey matter volumes, lateral ventricle volumes and total intracranial volume (ICV) in MDD patients with suicidal symptoms (N=451) vs healthy controls (N=1996) or MDD patients with no suicidal symptoms (N=650). MDD patients reporting suicidal plans or attempts showed a smaller ICV (P=4.12 × 10-3) or a 2.87% smaller volume compared with controls (Cohen's d=-0.284). In addition, we observed a nonsignificant trend in which MDD cases with suicidal symptoms had smaller subcortical volumes and larger ventricular volumes compared with controls. Finally, no significant differences (P=0.28-0.97) were found between MDD patients with and those without suicidal symptoms for any of the brain volume measures. This is by far the largest neuroimaging meta-analysis of suicidal behaviour in MDD to date. Our results did not replicate previous reports of association between subcortical brain structure and suicidality and highlight the need for collecting better-powered imaging samples and using improved suicidality assessment instruments.


Gut Microbiota Mediates the Protective Effects of Dietary Capsaicin against Chronic Low-Grade Inflammation and Associated Obesity Induced by High-Fat Diet.
Metabolic endotoxemia originating from dysbiotic gut microbiota has been identified as a primary mediator for triggering the chronic low-grade inflammation (CLGI) responsible for the development of obesity. Capsaicin (CAP) is the major pungent bioactivator in chili peppers and has potent anti-obesity functions, yet the mechanisms linking this effect to gut microbiota remain obscure. Here we show that mice fed a high-fat diet (HFD) supplemented with CAP exhibit lower levels of metabolic endotoxemia and CLGI associated with lower body weight gain. High-resolution responses of the microbiota were examined by 16S rRNA sequencing, short-chain fatty acid (SCFA) measurements, and phylogenetic reconstruction of unobserved states (PICRUSt) analysis. The results showed, among others, that dietary CAP induced increased levels of butyrate-producing Ruminococcaceae and Lachnospiraceae, while it caused lower levels of members of the lipopolysaccharide (LPS)-producing family S24_7. Predicted function analysis (PICRUSt) showed depletion of genes involved in bacterial LPS synthesis in response to CAP. We further identified that inhibition of cannabinoid receptor type 1 (CB1) by CAP also contributes to prevention of HFD-induced gut barrier dysfunction. Importantly, fecal microbiota transplantation experiments conducted in germfree mice demonstrated that dietary CAP-induced protection against HFD-induced obesity is transferrable. Moreover, microbiota depletion by a cocktail of antibiotics was sufficient to block the CAP-induced protective phenotype against obesity, further suggesting the role of microbiota in this context. Together, our findings uncover an interaction between dietary CAP and gut microbiota as a novel mechanism for the anti-obesity effect of CAP acting through prevention of microbial dysbiosis, gut barrier dysfunction, and chronic low-grade inflammation.IMPORTANCE Metabolic endotoxemia due to gut microbial dysbiosis is a major contributor to the pathogenesis of chronic low-grade inflammation (CLGI), which primarily mediates the development of obesity. A dietary strategy to reduce endotoxemia appears to be an effective approach for addressing the issue of obesity. Capsaicin (CAP) is the major pungent component in red chili (genus Capsicum). Little is known about the role of gut microbiota in the anti-obesity effect of CAP. High-throughput 16S rRNA gene sequencing revealed that CAP significantly increased butyragenic bacteria and decreased LPS-producing bacteria (e.g., members of the S24-7 family) and LPS biosynthesis. By using antibiotics and microbiota transplantation, we prove that gut microbiota plays a causal role in dietary CAP-induced protective phenotype against high-fat-diet-induced CLGI and obesity. Moreover, CB1 inhibition was partially involved in the beneficial effect of CAP. Together, these data suggest that the gut microbiome is a critical factor for the anti-obesity effects of CAP.


Predicting pregnancy status from mid-infrared spectroscopy in dairy cow milk using deep learning.
Accurately identifying pregnancy status is imperative for a profitable dairy enterprise. Mid-infrared (MIR) spectroscopy is routinely used to determine fat and protein concentrations in milk samples. Mid-infrared spectra have successfully been used to predict other economically important traits, including fatty acid content, mineral content, body energy status, lactoferrin, feed intake, and methane emissions. Machine learning has been used in a variety of fields to find patterns in vast quantities of data. This study aims to use deep learning, a sub-branch of machine learning, to establish pregnancy status from routinely collected milk MIR spectral data. Milk spectral data were obtained from National Milk Records (Chippenham, UK), who collect large volumes of data continuously on a monthly basis. Two approaches were followed: using genetic algorithms for feature selection and network design (model 1), and transfer learning with a pretrained DenseNet model (model 2). Feature selection in model 1 showed that the number of wave points in MIR data could be reduced from 1,060 to 196 wave points. The trained model converged after 162 epochs with validation accuracy and loss of 0.89 and 0.18, respectively. Although the accuracy was sufficiently high, the loss (in terms of predicting only 2 labels) was considered too high and suggested that the model would not be robust enough to apply to industry. Model 2 was trained in 2 stages of 100 epochs each with spectral data converted to gray-scale images and resulted in accuracy and loss of 0.97 and 0.08, respectively. Inspection on inference data showed prediction sensitivity of 0.89, specificity of 0.86, and prediction accuracy of 0.88. Results indicate that milk MIR data contains features relating to pregnancy status and the underlying metabolic changes in dairy cows, and such features can be identified by means of deep learning. Prediction equations from trained models can be used to alert farmers of nonviable pregnancies as well as to verify conception dates.


Characteristics, in-hospital and long-term clinical outcomes of nonagenarian compared with octogenarian acute myocardial infarction patients.
We compared clinical characteristics, management, and clinical outcomes of nonagenarian acute myocardial infarction (AMI) patients (n=270, 92.3 ± 2.3 yr old) with octogenarian AMI patients (n=2,145, 83.5 ± 2.7 yr old) enrolled in Korean AMI Registry (KAMIR). Nonagenarians were less likely to have hypertension, diabetes and less likely to be prescribed with beta-blockers, statins, and glycoprotein IIb/IIIa inhibitors compared with octogenarians. Although percutaneous coronary intervention (PCI) was preferred in octogenarians than nonagenarians, the success rate of PCI between the two groups was comparable. In-hospital mortality, the composite of in-hospital adverse outcomes and one year mortality were higher in nonagenarians than in octogenarians. However, the composite of the one year major adverse cardiac events (MACEs) was comparable between the two groups without differences in MI or re-PCI rate. PCI improved 1-yr mortality (adjusted hazard ratio [HR], 0.50; 95% confidence interval [CI], 0.36-0.69, P<0.001) and MACEs (adjusted HR, 0.47; 95% CI, 0.37-0.61, P<0.001) without significant complications both in nonagenarians and octogenarians. In conclusion, nonagenarians had similar 1-yr MACEs rates despite of higher in-hospital and 1-yr mortality compared with octogenarian AMI patients. PCI in nonagenarian AMI patients was associated to better 1-yr clinical outcomes.


Na-P(i) cotransport sites in proximal tubule and collecting tubule of winter flounder (Pleuronectes americanus).
Localization of a recently described and cloned Na-Pi cotransport system from flounder was investigated by reverse transcription-polymerase chain reaction (RT-PCR) of microdissected tubules and by immunocytochemistry of kidney of winter flounder. Histological examination showed a small glomerulus, an extremely short proximal tubule PI with a selective affinity to Lens culinaris agglutinin from lentils, and an extensive second proximal tubule segment PII (> 90% of proximal tubules), consisting of cells with numerous apical clear vesicles and extensive amplification of basolateral cell membranes. PII merged with the collecting tubule/ collecting duct (CT/CD) system without a distal segment. By RT-PCR, PII cells revealed high levels of NaPi-II related RNA; low levels were also observed in CTs. Previously characterized antisera against different epitopes of flounder NaPi-II specifically labeled the basolateral regions of PII and the apical cell portion of CT/CD cells and of some PII cells. These results suggest that tubular secretion of P(i) occurs in PII of teleost fish with modulation of urinary P(i) content in the subsequent CT/CD system.


Wnt up your mind - intervention strategies for S100A4-induced metastasis in colon cancer.
Colon cancer is still a burden mainly due to metastasis formation. The latter is often associated with a constitutive activation of the Wnt/beta-catenin signaling pathway and high expression of the metastasis-inducing gene S100A4. We previously identified S100A4 as a transcriptional target of beta-catenin. Intervention strategies targeting Wnt/beta-catenin signaling might therefore represent promising approaches to inhibit tumor growth and metastasis formation when induced by S100A4. Many inhibitors, various strategies, as well as different routes of application targeting key molecules of the Wnt signaling pathway have been reported within the last decade. Consequently, downregulation of beta-catenin target genes lead to altered tumorigenic and metastastic abilities of cancer cells. This review focuses on the potential of Wnt/beta-catenin signaling intervention to restrict colon cancer metastasis formation by interdicting S100A4 expression.


Legislation to Increase Uptake of HPV Vaccination and Adolescent Sexual Behaviors.
Despite preventive health benefits of the human papillomavirus (HPV) vaccination, uptake in the United States remains low. Twenty-four states have enacted legislation regarding HPV vaccination and education. One reason these policies have been controversial is because of concerns that they encourage risky adolescent sexual behaviors. Our aim in this study is to determine if state HPV legislation is associated with changes in adolescent sexual behaviors. This is a difference-in-difference study in which we use data on adolescent sexual behaviors from the school-based state Youth Risk Behavior Surveillance System from 2001 to 2015. Sexual behaviors included ever having sexual intercourse in the last 3 months and condom use during last sexual intercourse. We compared changes in sexual behaviors among high school students before and after HPV legislation to changes among high school students in states without legislation. A total of 715 338 participants reported ever having sexual intercourse in the last 3 months, and 217 077 sexually active participants reported recent condom use. We found no substantive or statistically significant associations between HPV legislation and adolescent sexual behaviors. Recent sexual intercourse decreased by 0.90 percentage points (P = .21), and recent condom use increased by 0.96 percentage points (P = .32) among adolescents in states that enacted legislation compared with states that did not. Results were robust to a number of sensitivity analyses. Implementation of HPV legislation was not associated with changes in adolescent sexual behaviors in the United States. Concern that legislation will increase risky adolescent sexual behaviors should not be used when deciding to pass HPV legislation.