Long-term follow-up of participants with heart failure in the antihypertensive and lipid-lowering treatment to prevent heart attack trial (ALLHAT).
In the Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial (ALLHAT), a randomized, double-blind, practice-based, active-control, comparative effectiveness trial in high-risk hypertensive participants, risk of new-onset heart failure (HF) was higher in the amlodipine (2.5-10 mg/d) and lisinopril (10-40 mg/d) arms compared with the chlorthalidone (12.5-25 mg/d) arm. Similar to other studies, mortality rates following new-onset HF were very high (≥50% at 5 years), and were similar across randomized treatment arms. After the randomized phase of the trial ended in 2002, outcomes were determined from administrative databases. With the use of national databases, posttrial follow-up mortality through 2006 was obtained on participants who developed new-onset HF during the randomized (in-trial) phase of ALLHAT. Mean follow-up for the entire period was 8.9 years. Of 1761 participants with incident HF in-trial, 1348 died. Post-HF all-cause mortality was similar across treatment groups, with adjusted hazard ratios (95% confidence intervals) of 0.95 (0.81-1.12) and 1.05 (0.89-1.25), respectively, for amlodipine and lisinopril compared with chlorthalidone, and 10-year adjusted rates of 86%, 87%, and 83%, respectively. All-cause mortality rates were also similar among those with reduced ejection fractions (84%) and preserved ejection fractions (81%), with no significant differences by randomized treatment arm. Once HF develops, risk of death is high and consistent across randomized treatment groups. Measures to prevent the development of HF, especially blood pressure control, must be a priority if mortality associated with the development of HF is to be addressed. Clinical Trial Registration- http://www.clinicaltrials.gov. Unique identifier: NCT00000542.


The coexistence of primary hyperparathyroidism and thyroid nodules: should the preoperative work-up of the parathyroid and the thyroid diseases be specifically adjusted?
Primary hyperparathyroidism (PHPT) can be found in concomitance with thyroid disease (TD) in a high frequency of cases. In this context the diagnostic exams for localizing the enlarged parathyroid(s) gland(s) could be less reliable or nonconclusive. Moreover, the thyroid carcinoma seems to be more frequent compared to that isolated thyroid desease and, therefore, carefully investigated. The main goal of the present study is to evaluate which diagnostic tool (US, MIBI) is more reliable for localizing the site of the PTH hypersecretion and to confirm if it is always advantageous a combination of both exams. Besides, we evaluated the incidence of thyroid carcinoma in our series of patients. A review of available data of 73 patients who underwent total thyroidectomy + parathyroidectomy from 2003 and 2014 was performed. The preoperative workup included systematically US and MIBI whose results were considered true positive when at least the side (left/right) of the parathyroid affected were concordant with the surgical report, settled as the gold standard, according to the Cox nonnested model. The connection between the diagnostic results of US versus MIBI was calculated with the Cohen K index for evaluating their overlap. The average of the thyroid carcinoma were also calculated. The difference between respectively US versus surgical report (p value=0.73) and MIBI versus surgical report (p value=0.81) were not significant. The low Cohen K index showed that both US and MIBI are complementary. In 23 patients (32,9%) a thyroid carcinoma was found. The association of MIBI and neck US is mandatory in the first evaluation of patients undergoing thyroidectomy and parathyroid excision simultaneously. The high prevalence of thyroid carcinoma in this specific context suggests a more aggressive diagnostic and surgical behaviour.


Carrier proteins-based boron delivery to tumor.
The conjugation of maleimide-functionalized closo-dodecaborate (MID) to transferrin (TF), which has no free SH cysteine residue, proceeded at room temperature for 12 h in PBS buffer (pH = 7.4). MID-TF conjugates were accumulated into the cells in the time- and concentration-dependent manners through the TF receptor mediated mechanism. In vivo biodistribution study of MID-TF conjugates in colon 26-bearing mice revealed the time-dependent selective accumulation of MID-TF conjugates into tumor very similar to that of MID-BSA. The tumor boron concentration of MID-BSA was higher than that of MID-TF conjugates (62 ppm vs. 20 ppm at a dose of 30 mg [B]/kg) 12 h after administration, suggesting that the EPR effect is not remarkably observed in the case of MID-TF conjugates because TF has smaller molecular weight than BSA (48 kDa vs. 68 kDa).


Tuberculosis and nutrition: disease perceptions and health seeking behavior of household contacts in the Peruvian Amazon.
Households of TB patients in the Peruvian Amazon. To investigate how knowledge and beliefs of household contacts about TB affected health seeking behavior. Interviews with 73 patients finishing treatment and 79 of their adult household contacts. Contacts were knowledgeable about free screening and treatment, but contacts who noted weight loss, not cough, were more likely to be screened for TB (P = 0.03). Forty-two per cent reported that TB was prevented by nutrition, 28% by separating eating utensils, and only 19% by avoiding a coughing patient. Only one household contact reported being stigmatized. Stigma centered upon nutrition, and only 12% knew of the association between TB and HIV. Only 14% had a BMI < 20, yet 30% reported regularly going to sleep hungry. Free food packages were reported to be the most important reason for treatment adherence by 33% of patients. Contacts misperceived TB as a nutritional disease and did not fear airborne transmission, which should be corrected by public health education. Weight loss, and not cough, led to screening. Stigma appeared to be minimized because risk was perceived as personal, through malnutrition, rather than exposure-based. Nutritional incentives that utilize these beliefs may reduce diagnostic delay and enhance treatment adherence.


Differential association of postsynaptic signaling protein complexes in striatum and hippocampus.
Distinct physiological stimuli are required for bidirectional synaptic plasticity in striatum and hippocampus, but differences in the underlying signaling mechanisms are poorly understood. We have begun to compare levels and interactions of key excitatory synaptic proteins in whole extracts and subcellular fractions isolated from micro-dissected striatum and hippocampus. Levels of multiple glutamate receptor subunits, calcium/calmodulin-dependent protein kinase II (CaMKII), a highly abundant serine/threonine kinase, and spinophilin, a F-actin and protein phosphatase 1 (PP1) binding protein, were significantly lower in striatal extracts, as well as in synaptic and/or extrasynaptic fractions, compared with similar hippocampal extracts/fractions. However, CaMKII interactions with spinophilin were more robust in striatum compared with hippocampus, and this enhanced association was restricted to the extrasynaptic fraction. NMDAR GluN2B subunits associate with both spinophilin and CaMKII, but spinophilin-GluN2B complexes were enriched in extrasynaptic fractions whereas CaMKII-GluN2B complexes were enriched in synaptic fractions. Notably, the association of GluN2B with both CaMKII and spinophilin was more robust in striatal extrasynaptic fractions compared with hippocampal extrasynaptic fractions. Selective differences in the assembly of synaptic and extrasynaptic signaling complexes may contribute to differential physiological regulation of excitatory transmission in striatum and hippocampus.


Novel CSF biomarkers to discriminate FTLD and its pathological subtypes.
Frontotemporal lobar degeneration (FTLD) is the second most prevalent dementia in young patients and is characterized by the presence of two main protein aggregates in the brain, tau (FTLD-Tau) or TDP43 (FTLD-TDP), which likely require distinct pharmacological therapy. However, specific diagnosis of FTLD and its subtypes remains challenging due to largely overlapping clinical phenotypes. Here, we aimed to assess the clinical performance of novel cerebrospinal fluid (CSF) biomarkers for discrimination of FTLD and its pathological subtypes. YKL40, FABP4, MFG-E8, and the activities of catalase and specific lysosomal enzymes were analyzed in patients with FTLD-TDP (n = 30), FTLD-Tau (n = 20), AD (n = 30), DLB (n = 29), and nondemented controls (n = 29) obtained from two different centers. Models were validated in an independent CSF cohort (n = 188). YKL40 and catalase activity were increased in FTLD-TDP cases compared to controls. YKL40 levels were also higher in FTLD-TDP compared to FTLD-Tau. We identified biomarker models able to discriminate FTLD from nondemented controls (MFG-E8, tTau, and Aβ 42; 78% sensitivity and 83% specificity) and non-FTLD dementia (YKL40, pTau, p/tTau ratio, and age; 90% sensitivity, 78% specificity), which were validated in an independent cohort. In addition, we identified a biomarker model differentiating FTLD-TDP from FTLD-Tau (YKL40, MFGE-8, βHexA together with βHexA/tHex and p/tTau ratios and age) with 80% sensitivity and 82% specificity. This study identifies CSF protein signatures distinguishing FTLD and the two main pathological subtypes with optimal accuracy (specificity/sensitivity > 80%). Validation of these models may allow appropriate selection of cases for clinical trials targeting the accumulation of Tau or TDP43, thereby increasing their efficiency and facilitating the development of successful therapies.


Accurate method for determining the refractive-index profiles of planar waveguides in uniaxial media with the optical axis normal to the surface.
A new method for characterization of uniaxial planar waveguides from their measured effective mode indices is presented. The theory is outlined and expressions for efficient computer analysis are given. Uniaxial waveguide samples have been made in c-cut LiNbO(3) by proton exchange with and without post annealing in order to test the method on both steplike and graded-index profiles. The resulting characterization of the samples is discussed in relation to the inverse WKB method. Finally, the importance of incorporating the effects of material birefringence in the characterization of these kinds of waveguides is investigated.


Prenatal HIV testing in private OB-Gyn practices in Texas.
This study evaluates the prenatal human immunodeficiency virus (HIV) testing behaviors of private obstetrics and gynecology practitioners in Texas. A statewide telephone survey of 614 providers of prenatal care determined the level of HIV testing, how testing is offered, and patient acceptance of the test. Ninety-nine percent of the practices offered HIV testing to all their pregnant patients, and 96% of the practices included HIV testing in the routine panel of tests for pregnant patients. More than 95% of the practices reported that 10% or less of the women refused the test when offered it; 73% of the practices reported no refusals. Less than half of the practices, however, discussed HIV prevention topics, and only 29% of the practices referred high-risk pregnant patients for prevention counseling. Although private practices of obstetrics and gynecology report testing almost all their prenatal patients, survey results suggest that providers could improve their prevention and patient education practices.


Computational modeling of serum-binding proteins and clearance in extrapolations across life stages and species for endocrine active compounds.
One measure of the potency of compounds that lead to the effects through ligand-dependent gene transcription is the relative affinity for the critical receptor. Endocrine active compounds that are presumed to act principally through binding to the estrogen receptor (e.g., estradiol, genistein, bisphenol A, and octylphenol) comprise one class of such compounds. For making simple comparisons, receptor-binding affinity has been equated to in vivo potency, which consequently defines the dose-response characteristics for the compound. Direct extrapolation of in vitro estimated affinities to the corresponding in vivo system and to specific species or life stages (e.g., neonatal, pregnancy) can be misleading. Accurate comparison of the potency of endocrine active compounds requires characterization of biochemical and pharmacokinetic factors that affect their free concentration. Quantitative in vitro and in vivo models were developed for integrating pharmacokinetics factors (e.g., serum protein and receptor-binding affinities, clearance) that affect potency. Data for parameterizing these models for several estrogenic compounds were evaluated and the models exercised. While simulations of adult human or rat sera were generally successful, difficulties in describing early life stages were identified. Exogenous compounds were predicted to be largely ineffective at competing estradiol off serum-binding proteins, suggesting this was unlikely to be physiologically significant. Discrepancies were identified between relative potencies based upon modeling in vitro receptor-binding activity versus in vivo activity in the presence of clearance and serum-binding proteins. The examples illustrate the utility of this approach for integrating available experimental data from in vitro and in vivo studies to estimate the relative potency of these compounds.


All's well that ends well: alternative polyadenylation and its implications for stem cell biology.
Stem cell quiescence, activation, and differentiation are governed by a complex network of molecular pathways. There has been a growing recognition that posttranscriptional modifications, such as alternative polyadenylation (APA) of transcripts, play an important role in regulating gene expression and function. Recent analyses of stem cell populations have suggested that APA controls stem cell fate and behavior. Here, we review recent developments that have shaped our understanding of the control of stem cell behavior by APA and we highlight promising areas for future investigation.


Visual pathway neurodegeneration winged by mitochondrial dysfunction.
To test for structural and functional contribution of mitochondrial dysfunction to neurodegeneration in multiple sclerosis (MS). A visual pathway model void of MS lesions was chosen in order to exclude neurodegeneration secondary to lesion related axonotmesis. A single-centre cohort study (230 MS patients, 63 controls). Spectral domain optical coherence tomography of the retina, 3T magnetic resonance imaging of the brain, spectrophotometric assessment of serum lactate levels. Postmortem immunohistochemistry. The visual pathway was void of MS lesions in 31 patients and 31 age-matched controls. Serum lactate was higher in MS compared to controls (P = 0.029). High serum lactate was structurally related to atrophy of the retinal nerve fiber layer at the optic disc (P = 0.041), macula (P = 0.025), and the macular ganglion cell complex (P = 0.041). High serum lactate was functionally related to poor color vision (P < 0.01), Expanded Disability Status Scale score (R = 0.37, P = 0.041), Guy's Neurological disability score (R = 0.38, P = 0.037), MS walking scale (R = 0.50, P = 0.009), upper limb motor function (R = 0.53, P = 0.002). Immunohistochemistry demonstrated increased astrocytic expression of a key lactate generating enzyme in MS lesions as well as profound vascular expression of monocarboxylate transporter-1, which is involved in lactate transport. This study provides structural, functional, and translational evidence for visual pathway neurodegeneration in MS related to mitochondrial dysfunction.


Avoiding Drug Resistance by Substrate Envelope-Guided Design: Toward Potent and Robust HCV NS3/4A Protease Inhibitors.
Hepatitis C virus (HCV) infects millions of people worldwide, causing chronic liver disease that can lead to cirrhosis, hepatocellular carcinoma, and liver transplant. In the last several years, the advent of direct-acting antivirals, including NS3/4A protease inhibitors (PIs), has remarkably improved treatment outcomes of HCV-infected patients. However, selection of resistance-associated substitutions and polymorphisms among genotypes can lead to drug resistance and in some cases treatment failure. A proactive strategy to combat resistance is to constrain PIs within evolutionarily conserved regions in the protease active site. Designing PIs using the substrate envelope is a rational strategy to decrease the susceptibility to resistance by using the constraints of substrate recognition. We successfully designed two series of HCV NS3/4A PIs to leverage unexploited areas in the substrate envelope to improve potency, specifically against resistance-associated substitutions at D168. Our design strategy achieved better resistance profiles over both the FDA-approved NS3/4A PI grazoprevir and the parent compound against the clinically relevant D168A substitution. Crystallographic structural analysis and inhibition assays confirmed that optimally filling the substrate envelope is critical to improve inhibitor potency while avoiding resistance. Specifically, inhibitors that enhanced hydrophobic packing in the S4 pocket and avoided an energetically frustrated pocket performed the best. Thus, the HCV substrate envelope proved to be a powerful tool to design robust PIs, offering a strategy that can be translated to other targets for rational design of inhibitors with improved potency and resistance profiles.IMPORTANCE Despite significant progress, hepatitis C virus (HCV) continues to be a major health problem with millions of people infected worldwide and thousands dying annually due to resulting complications. Recent antiviral combinations can achieve >95% cure, but late diagnosis, low access to treatment, and treatment failure due to drug resistance continue to be roadblocks against eradication of the virus. We report the rational design of two series of HCV NS3/4A protease inhibitors with improved resistance profiles by exploiting evolutionarily constrained regions of the active site using the substrate envelope model. Optimally filling the S4 pocket is critical to avoid resistance and improve potency. Our results provide drug design strategies to avoid resistance that are applicable to other quickly evolving viral drug targets.


Efficacy of telephone counseling for pregnant smokers: a randomized controlled trial.
Reducing tobacco use in pregnancy is a public health priority. Brief smoking counseling during prenatal care is effective but generates modest cessation rates. Telephone counseling is an effective smoking cessation method that could offer pregnant women convenient access to more intensive smoking cessation counseling. The efficacy of proactive pregnancy-tailored telephone counseling for smoking cessation was compared with a "best-practice" brief-counseling control in a randomized controlled trial of 442 pregnant smokers referred by prenatal providers and a managed care plan. Trained counselors using cognitive-behavioral and motivational interviewing methods called intervention subjects throughout pregnancy and for 2 months postpartum (mean = 5 calls, mean total contact = 68 minutes). Controls received one 5-minute counseling call. Cotinine-validated 7-day tobacco abstinence rates in intervention and control groups were 10.0% and 7.5% at end of pregnancy (odds ratio [OR] 1.37, 95% confidence interval [CI] 0.69-2.70; number needed to treat = 40) and 6.7% versus 7.1% at 3 months postpartum (OR 0.93, 95% CI 0.44-1.99). The intervention increased end-of-pregnancy cessation rates among 201 light smokers (< 10 cigarettes/day at study enrollment) (intervention 19.1% versus control 8.4%; OR 2.58, 95% CI 1.1-6.1; number needed to treat = 9.3) and among 193 smokers who attempted to quit in pregnancy before enrollment (intervention 18.1% versus control 6.8%; OR 3.02, CI 1.15-7.94; number needed to treat = 8.8); 63% of the sample (n = 267) was in one of these subgroups. Proactive pregnancy-tailored telephone counseling did not outperform a brief "best practice" intervention among pregnant smokers. The intervention had efficacy in light smokers and in women who had attempted cessation earlier in pregnancy. Future studies should confirm whether telephone counseling benefits these groups of pregnant smokers. ClinicalTrials.gov, #NCT00181909. I.


Effect of prevention of inorganic sulfate depletion on the pharmacokinetics of acetaminophen in rats.
The elimination of large doses of acetaminophen is associated with substantial depletion of endogenous inorganic sulfate which is utilized for the formation of acetaminophen sulfate. This depletion has pronounced dose- and time-dependent effects on the pharmacokinetics of acetaminophen. The purposes of this investigation were to determine the pharmacokinetics of acetaminophen in rats when endogenous sulfate depletion is prevented by administration of inorganic sulfate and to develop a simple, physiologically based pharmacokinetic model for the elimination of acetaminophen under these conditions. Adult Sprague-Dawley rats received an i.v. injection and a continuous infusion of sodium sulfate as well as an i.v. injection of acetaminophen, either 15, 30, 150 or 300 mg/kg. Serum inorganic sulfate concentrations remained at or above the physiologic level at all times. Plasma concentrations of acetaminophen declined exponentially with time after the two lower doses but exhibited initial downward curvature in log-linear plots after the two larger doses. The time-averaged plasma clearance of acetaminophen decreased with increasing dose whereas the terminal half-life was dose-independent. Most of the drug was eliminated in the urine as acetaminophen sulfate but the dose fractions of acetaminophen glucuronide and unmetabolized drug excreted in the urine increased with increasing dose. The renal clearance of acetaminophen did not exhibit dose-dependence but the apparent formation clearance of acetaminophen glucuronide tended to decrease with increasing dose. The formation of acetaminophen sulfate is describable by Michaelis-Menten kinetics, with a Vmax of about 6.5 mumol/min/kg and an in vivo KM (referenced to plasma) of about 100 microM.(ABSTRACT TRUNCATED AT 250 WORDS)


Intra-operative tissue oxygen tension is increased by local insufflation of humidified-warm CO2 during open abdominal surgery in a rat model.
Maintenance of high tissue oxygenation (PtO2) is recommended during surgery because PtO2 is highly predictive of surgical site infection and colonic anastomotic leakage. However, surgical site perfusion is often sub-optimal, creating an obstructive hurdle for traditional, systemically applied therapies to maintain or increase surgical site PtO2. This research tested the hypothesis that insufflation of humidified-warm CO2 into the abdominal cavity would increase sub-peritoneal PtO2 during open abdominal surgery. 15 Wistar rats underwent laparotomy under general anesthesia. Three sets of randomized cross-over experiments were conducted in which the abdominal cavity was subjected to alternating exposure to 1) humidified-warm CO2 & ambient air; 2) humidified-warm CO2 & dry-cold CO2; and 3) dry-cold CO2 & ambient air. Sub-peritoneal PtO2 and tissue temperature were measured with a polarographic oxygen probe. Upon insufflation of humidified-warm CO2, PtO2 increased by 29.8 mmHg (SD 13.3; p<0.001), or 96.6% (SD 51.9), and tissue temperature by 3.0°C (SD 1.7 p<0.001), in comparison with exposure to ambient air. Smaller, but significant, increases in PtO2 were seen in experiments 2 and 3. Tissue temperature decreased upon exposure to dry-cold CO2 compared with ambient air (-1.4°C, SD 0.5, p = 0.001). In a rat model, insufflation of humidified-warm CO2 into the abdominal cavity during open abdominal surgery causes an immediate and potentially clinically significant increase in PtO2. The effect is an additive result of the delivery of CO2 and avoidance of evaporative cooling via the delivery of the CO2 gas humidified at body temperature.


Effect of radiotherapy interruptions on survival in medicare enrollees with local and regional head-and-neck cancer.
To investigate whether interruptions in radiotherapy are associated with decreased survival in a population-based sample of head-and-neck cancer patients. Using the Surveillance, Epidemiology, and End Results-Medicare linked database we identified Medicare beneficiaries aged 66 years and older diagnosed with local-regional head-and-neck cancer during the period 1997-2003. We examined claims records of 3864 patients completing radiotherapy for the presence of one or more 5-30-day interruption(s) in therapy. We then performed Cox regression analyses to estimate the association between therapy interruptions and survival. Patients with laryngeal tumors who experienced an interruption in radiotherapy had a 68% (95% confidence interval, 41-200%) increased risk of death, compared with patients with no interruptions. Patients with nasal cavity, nasopharynx, oral, salivary gland, and sinus tumors had similar associations between interruptions and increased risk of death, but these did not reach statistical significance because of small sample sizes. Treatment interruptions seem to influence survival time among patients with laryngeal tumors completing a full course of radiotherapy. At all head-and-neck sites, the association between interruptions and survival is sensitive to confounding by stage and other treatments. Further research is needed to develop methods to identify patients most susceptible to interruption-induced mortality.


Rationale and design of the Investigation of the Management of Pericarditis (IMPI) trial: a 2 × 2 factorial randomized double-blind multicenter trial of adjunctive prednisolone and Mycobacterium w immunotherapy in tuberculous pericarditis.
In spite of antituberculosis chemotherapy, tuberculous (TB) pericarditis causes death or disability in nearly half of those affected. Attenuation of the inflammatory response in TB pericarditis may improve outcome by reducing cardiac tamponade and pericardial constriction, but there is uncertainty as to whether adjunctive immunomodulation with corticosteroids and Mycobacterium w (M. w) can safely reduce mortality and morbidity. The primary objective of the IMPI Trial is to assess the effectiveness and safety of prednisolone and M. w immunotherapy in reducing the composite outcome of death, constriction, or cardiac tamponade requiring pericardial drainage in 1,400 patients with TB pericardial effusion. The IMPI trial is a multicenter international randomized double-blind placebo-controlled 2 × 2 factorial study. Eligible patients are randomly assigned to receive oral prednisolone or placebo for 6 weeks and M. w injection or placebo for 3 months. Patients are followed up at weeks 2, 4, and 6 and months 3 and 6 during the intervention period and 6-monthly thereafter for up to 4 years. The primary outcome is the first occurrence of death, pericardial constriction, or cardiac tamponade requiring pericardiocentesis. The secondary outcome is safety of immunomodulatory treatment measured by effect on opportunistic infections (eg, herpes zoster) and malignancy (eg, Kaposi sarcoma) and impact on measures of immunosuppression and the incidence of immune reconstitution disease. IMPI is the largest trial yet conducted comparing adjunctive immunotherapy in pericarditis. Its results will define the role of adjunctive corticosteroids and M. w immunotherapy in patients with TB pericardial effusion.


Coated microneedles for drug delivery to the eye.
To test the hypothesis that coated microneedles can deliver drugs into the eye via intrascleral and intracorneal routes in a minimally invasive manner. Solid metal microneedles measuring 500 to 750 microm in length were coated with model drugs, protein, and DNA; inserted into nonpreserved human cadaveric sclera; and imaged. Microneedles coated with sodium fluorescein were then inserted into rabbit cornea in vivo. After needle removal, fluorescein concentration in the anterior segment of the rabbit eye was measured for 24 hours. Similar experiments were performed using pilocarpine-coated microneedles, and the rabbit pupil size was monitored afterward. In vitro insertion tests showed that microneedles were mechanically strong enough to penetrate into human cadaveric sclera and that the drug coating rapidly dissolved off the needles within the scleral tissue within 30 seconds after insertion. In vivo delivery from fluorescein-coated microneedles showed that fluorescein concentrations in the anterior chamber were 60 times greater than those achieved by topical application without microneedles. Similarly, microneedle delivery of pilocarpine caused rapid and extensive rabbit pupil constriction. There were no measurable inflammatory responses caused by microneedle insertion. This study demonstrated for the first time that coated microneedles can deliver drugs into the eye via intrascleral and intracorneal routes. This minimally invasive approach may avoid the complications associated with intraocular injection and systemic administration.


Fluoride and arsenic contamination in drinking water due to mining activities and its impact on local area population.
Contamination of arsenic and fluoride in drinking water reservoirs is a serious health issue in the Sibi district, Balochistan, Pakistan. The contamination has already been affecting a large population of the district. Dental fluorosis and dermatitis are the most common reported illnesses in the area. This study focused on the evaluation of the root causes and pathway by which it reached to the body. Questioner analysis, simple examination, and pictorial representation were used to study the prevalence of diseases caused by As and F. People of the Sibi district were found to consume both surface and groundwater, which were highly contaminated with arsenic and fluoride. The saturation index, ranging from 1 to 7, showed high enrichment of contaminants in both types of water. Geochemical calculations and the sodium absorption ratio were evaluated. High values of the saturation index of different salts showed high saturation of salts in water. The principal component analysis grouped the data into three clusters, showing that the surface water has no resemblance with the control water. High degree of contamination was observed for most of the samples, whereas, some samples of ground water were closed to the control group; a group of samples within WHO limits. The correlation studies and other calculations also revealed that the F and As reached lethal limits in the drinking water and thus caused severe health damage to the local area population. The diseases found in the area are fluorosis, keratosis, dermatitis, and melanosis.


Eye care providers' attitudes towards tele-ophthalmology.
The rapid rise of e-health and remote care systems will likely change the practice patterns of ophthalmologists. Although telemedicine practices are thriving in many specialties of medicine, telemedicine for ophthalmology has been limited primarily to asynchronous care for diabetic retinopathy. The goal of this research was to evaluate perspectives on and familiarity with telemedicine among eye care providers at a large tertiary-care medical center via an anonymous, descriptive survey. In total, 58 eye care physicians completed surveys (response rates of 86% for physicians-in-training and 49% for faculty physicians, respectively). Although a majority of both faculty and physicians-in-training were willing to participate in telemedicine services, trainees were more likely to be willing to interpret photographs than faculty (p=0.04). Most respondents (71%) indicated that they did not use telemedicine. Over half had received photographs (via phone or e-mail) for interpretation from referring physicians (54%) or patients (56%) within the past 3 months. A majority of providers (82%) would be willing to participate in telemedicine for consultations and for interpreting photographs, but a majority (59%) had low confidence in remote care for providing an opinion on patient care. Most eye care providers viewed telemedicine as part of the future of eye care but were concerned about the use of telemedicine. Although most providers did not practice telemedicine, over half of them were comfortable managing eye care consultations (including patients' photographs) via the Internet.


Endometriosis in association with Herlyn-Werner-Wunderlich syndrome.
To describe the clinical characteristics of patients with Herlyn-Werner-Wunderlich syndrome (HWWS) and explore the relationship between an obstructive genital abnormality and pelvic endometriosis. Case series. University hospital. A total of 94 women with Herlyn-Werner-Wunderlich syndrome (HWWS). Retrospectively reviewed charts of patients diagnosed with HWWS and follow-up contact. Data for each patient regarding demographics, presenting symptoms, concomitant complications, and anatomic variations were obtained from operation notes and clinical records. The occurrence of pelvic endometriosis was 19.15% (18/94) in patients with HWWS. All ovarian endometrial cysts (100%) occurred ipsilateral to the vaginal septum. Pelvic endometriosis was significantly more frequent in patients with complete hemivaginal obstructions (10/27, 37.0%) than those with incomplete obstructions (8/67, 11.9%). One-fifth of patients with HWWS were susceptible to pelvic endometriosis, and all of the ovarian endometriotic cysts were ipsilateral to the vaginal septum. Pelvic endometriosis in adolescents appeared to be related to the obstructed genital abnormality. Vaginal septum resection should be the first step in treatment and surgery has an important role in the treatment of endometriosis and pelvic adhesion.


Planning Implant Placement on 3D Stereolithographic Models Applied with Immediate Loading of Implant-Supported Hybrid Prostheses After Multiple Extractions: A Case Series.
The aim of this noninterventional, retrospective case series was to evaluate the outcome of immediately loaded implants in patients with failing dentitions that require bone tabling using a bone reduction guide and a surgical guide manufactured directly on three-dimensional (3D) stereolithographic models. Consecutive patients with failing dentition and at least two remaining teeth who were treated in a single center between December 14, 2009 and September 23, 2013 were eligible. All patients receiving implants loaded with a hybrid prosthesis on the same day as extraction with their surgery planned on 3D models and performed using a surgical guide manufactured in a laboratory on the planning model were included. Patients who had undergone bone grafting procedures were excluded. Descriptive statistical analyses of available data were performed, including life-table calculations to derive a cumulative survival rate (CSR). Two hundred twenty-eight patients (105 females and 123 males) received 1,657 implants (NobelActive) in 321 arches, in most cases 5 implants (range, 5 to 7) per arch. Ten preexisting implants were used. The mean insertion torque was 60.02 ± 13.1 Ncm (range, 15 to 75 Ncm). The definitive abutment was placed on the same day as surgery in all cases, and the definitive prosthesis (n = 304) was delivered after a mean of 7.9 ± 2.6 months. All implants were followed for 20.01 ± 11.3 months (range, 0 to 52 months) from implant insertion. Four implants (3 patients) had delayed loading, and one implant was left as a sleeping implant. Eight implants among six patients failed, two of the implants after prosthesis delivery. The CSR of the placed implants was 99.4% at implant level and 96.2% at patient level. Planning on 3D models to remove bone and place implants using custom-made bone reduction and surgical guides with immediate loading on the same day as extraction of remaining teeth was safe and effective for implant survival and rehabilitation of patients with periodontitis and failing dentition.


Influence of Sleep Dysfunction on Concussion Assessment Outcomes Among Adolescent Athletes After Concussion and Healthy Controls.
Sleep dysfunction (SD) is associated with a high symptom burden and lower neurocognitive performance after concussion and on baseline testing without injury. However, few studies have compared concussed athletes and controls with and without SD on clinical outcomes. To evaluate differences in clinical outcomes among both concussed athletes and matched controls with and without SD. Retrospective cross-sectional study. Participants aged 12 to 20 years were recruited from a concussion clinic (n = 50 patients) and research registry/flyers (n = 50 healthy age-/sex-matched controls). Participants were categorized by self-reported SD into one of 4 groups: sport-related concussion (SRC) + SD, SRC only, SD only, and controls. Post-Concussion Symptom Scale (PCSS), Vestibular/Oculomotor Screening (VOMS), and neurocognitive testing (Immediate Postconcussion Assessment Cognitive Test). Compared with the SRC only group, the SRC + SD group performed worse on all neurocognitive domains, had a higher total symptom score, and endorsed more symptoms on most VOMS items. In addition, the SRC + SD group was at an increased likelihood of having at least 1 abnormal VOMS item compared with SRC only group. The SRC only group had neurocognitive test scores and symptom reports statistically similar to the SD only group. Sleep dysfunction after concussion is related to worse neurocognitive performance and higher concussion symptom reporting. This study extended findings to suggest vestibular symptomology is worse among athletes with SD after injury compared to injured athletes without SD. Similar performances on concussion assessments for the SRC only and SD only groups suggest SD may appear similar to clinical presentation of concussion, even at baseline in the absence of SRC.


Katrina in Historical Context: Environment and Migration in the U.S.
The massive publicity surrounding the exodus of residents from New Orleans spurred by Hurricane Katrina has encouraged interest in the ways that past migration in the U.S. has been shaped by environmental factors. So has Timothy Egan's exciting book, The Worst Hard Time: The Untold Story of those who survived the Great American Dust Bowl. This paper places those dramatic stories into a much less exciting context, demonstrating that the kinds of environmental factors exemplified by Katrina and the Dust Bowl are dwarfed in importance and frequency by the other ways that environment has both impeded and assisted the forces of migration. We accomplish this goal by enumerating four types of environmental influence on migration in the U.S.: 1) environmental calamities, including floods, hurricanes, earthquakes, and tornadoes, 2) environmental hardships and their obverse, short-term environmental benefits, including both drought and short periods of favorable weather, 3) environmental amenities, including warmth, sun, and proximity to water or mountains, and 4) environmental barriers and their management, including heat, air conditioning, flood control, drainage, and irrigation. In U.S. history, all four of these have driven migration flows in one direction or another. Placing Katrina into this historical context is an important task, both because the environmental calamities of which Katrina is an example are relatively rare and have not had a wide impact, and because focusing on them defers interest from the other kinds of environmental impacts, whose effect on migration may have been stronger and more persistent, though less dramatic.


THE IMPORTANCE OF COLONOSCOPY IN INFLAMMATORY BOWEL DISEASES.
Endoscopic evaluation, particularly the macroscopic mucosal and histological results of ileocolic biopsies, is essential for the management of inflammatory bowel disease. Endoscopic appearance is not always sufficient to differentiate Crohn's disease and ulcerative colitis, but there are some characteristics that favor one or another diagnosis. Both diseases have an increased incidence of colorectal carcinoma; so, surveillance colonoscopy is important for detecting early neoplastic lesions. To update the importance of endoscopy in the evaluation, diagnosis and prognosis of inflammatory bowel disease. Search was done in the scientific literature of the TRIP database, chosen from clinical questions (PICO) with the following descriptors: "inflammatory bowel disease", "endoscopy/colonoscopy", "Crohn's disease", "ulcerative colitis" and "diagnosis/treatment". Endoscopic investigation in patients with chronic colitis is quite accurate for the differential diagnosis between ulcerative colitis and Crohn's disease. Endoscopy is indicated for ulcerative colitis during severe crisis due to its prognostic value. Another accepted indication for endoscopy in inflammatory bowel disease is its use in the screening for dysplastic lesion. Ileocolonoscopy allows an accurate diagnosis of Crohn's disease or ulcerative colitis in up to 90% of cases. The healing of the mucosa assessed by endoscopy after treatments despite not being consensus is still the gold-standard in the evaluation of remission of the disease. Colonoscopy is essential for long-term cancer surveillance and in the future the implementation of Confocal Laser Endomicroscopy seems to be very promising in assessing the initial dysplasia.


Is heart period variability associated with the administration of lifesaving interventions in individual prehospital trauma patients with normal standard vital signs?
To determine whether heart period variability provides added value in identifying the need for lifesaving interventions (LSI) in individual trauma patients with normal standard vital signs upon early medical assessment. Retrospective database review. Helicopter transport to Level 1 trauma center and first 24 hrs of in-hospital care. Prehospital trauma patients requiring helicopter transport to Level 1 trauma center. Heart period variability was analyzed from electrocardiographic recordings collected from 159 prehospital trauma patients with normal standard vital signs (32 LSI patients, 127 No-LSI patients). Although 13 of the electrocardiogram derived metrics demonstrated simple (i.e., univariate) discrimination between groups, at the multivariate level, only fractal dimension by curve length (FD-L) was uniquely associated with group membership (LSI vs. No-LSI, p = .0004). Whereas the area under the receiver operating characteristics curve for FD-L was 0.70, the overall correct classification rate (true positives and true negatives) of 82% was only 2% higher than the baseline prediction rate of 80% (i.e., no information except for the known proportion of overall No-LSI cases, 127 of 159 patients). Furthermore, 84% of the individual FD-L values for the LSI group were within the range of the No-LSI group. Only FD-L was uniquely able to distinguish patient groups based on mean values when standard vital signs were normal. However, the accuracy of FD-L in distinguishing between patients was only slightly better than the baseline prediction rate. There was also very high overlap of individual heart period variability values between groups, so many LSI patients could be incorrectly classified as not requiring an LSI if a single heart period variability value was used as a triage tool. Based on this analysis, heart period variability seems to have limited value for prediction of LSIs in prehospital trauma patients with normal standard vital signs.


Rapid task-dependent tuning of the mouse olfactory bulb.
Adapting neural representation to rapidly changing behavioural demands is a key challenge for the nervous system. Here, we demonstrate that the output of the primary olfactory area of the mouse, the olfactory bulb, is already a target of dynamic and reproducible modulation. The modulation depends on the stimulus tuning of a given neuron, making olfactory responses more discriminable through selective amplification in a demand-specific way.


Prolonged-release fampridine improves walking in a proportion of patients with multiple sclerosis.
Fampridine is indicated to improve walking in adult multiple sclerosis (MS) patients. Indications vary between countries and the prescribing neurologist should be aware of the labeling and indication in his own country. The prolonged-release formulation of 4-aminopyridine has reduced the risk of seizure to a level near the intrinsic MS risk, and the risk can be further minimized if it emphasized that patients should not exceed the recommended dose of 10 mg twice a day, should not catch up on missed doses and should not divide, crush or chew tablets. It is imperative to check the renal function before and during treatment and make sure the patient does not get concomitant medications affecting the renal elimination. The use of fampridine is considered safe, and the side effects are often mild and acceptable. Approximately one-third of MS patients treated with fampridine will experience an improvement of their walking speed above 20% on the timed 25-foot walk test (T25FW), which is considered to be clinically relevant.


Host plant defense produces species-specific alterations to flight muscle protein structure and flight-related fitness traits of two armyworms.
Insects manifest phenotypic plasticity in their development and behavior in response to plant defenses, via molecular mechanisms that produce tissue-specific changes. Phenotypic changes might vary between species that differ in their preferred hosts and these effects could extend beyond larval stages. To test this, we manipulated the diet of southern armyworm (SAW; Spodoptera eridania) and fall armyworm (FAW; Spodoptera frugiperda) using a tomato mutant for jasmonic acid plant defense pathway (def1), and wild-type plants, and then quantified gene expression of Troponin t (Tnt) and flight muscle metabolism of the adult insects. Differences in Tnt spliceform ratios in insect flight muscles correlate with changes to flight muscle metabolism and flight muscle output. We found that SAW adults reared on induced def1 plants had a higher relative abundance (RA) of the A isoform of Troponin t (Tnt A) in their flight muscles; in contrast, FAW adults reared on induced def1 plants had a lower RA of Tnt A in their flight muscles compared with adults reared on def1 and controls. Although mass-adjusted flight metabolic rate showed no independent host plant effects in either species, higher flight metabolic rates in SAW correlated with increased RA of Tnt A Flight muscle metabolism also showed an interaction of host plants with Tnt A in both species, suggesting that host plants might be influencing flight muscle metabolic output by altering Tnt This study illustrates how insects respond to variation in host plant chemical defense by phenotypic modifications to their flight muscle proteins, with possible implications for dispersal.


Genetic architectures of ADME genes in five Eurasian admixed populations and implications for drug safety and efficacy.
Drug absorption, distribution, metabolism and excretion (ADME) contribute to the high heterogeneity of drug responses in humans. However, the same standard for drug dosage has been applied to all populations in China although genetic differences in ADME genes are expected to exist in different ethnic groups. In particular, the ethnic minorities in northwestern China with substantial ancestry contribution from Western Eurasian people might violate such a single unified standard. In this study, we used Affymetrix SNP Array 6.0 to investigate the genetic diversity of 282 ADME genes in five northwestern Chinese minority populations, namely, Tajik, Uyghur, Kazakh, Kirgiz and Hui, and attempted to identify the highly differential SNPs and haplotypes and further explore their clinical implications. We found that genetic diversity of many ADME genes in the five minority groups was substantially different from those in the Han Chinese population. For instance, we identified 10 functional SNPs with substantial allele frequency differences, 14 functional SNPs with highly different heterozygous states and eight genes with significant haplotype differences between these admixed minority populations and the Han Chinese population. We further confirmed that these differences mainly resulted from the European gene flow, that is, this gene flow increased the genetic diversity in the admixed populations. These results suggest that the ADME genes vary substantially among different Chinese ethnic groups. We suggest it could cause potential clinical risk if the same dosage of substances (eg, antitumour drugs) is used without considering population stratification.


[Limnetic zooplankton run-off a high-head dam and their fate in a river with high current velocity (case of the Krasnoiarsk hydroelectric power station on the Yenisei river].
The vertical distribution of net zooplankton in head-water of Krasnoyarsk hydroelectric power station and its horizontal distribution in the tail-water were studied during two years in winter and summer seasons. In order to distinguish living and dead individuals the special staining was used. It was revealed that on average 77% of living plankton pass through high-head dam with deep water scoop to the tailwater. While passing through dam aggregates some individuals of the reservoir plankton are traumatized and die, that results in some increase of portion of dead individuals in the tail water near dam (from 3 to 6%). Alive zooplankton passed through the dam aggregates is eliminated under the Upper Yenisei highly turbulent conditions. There is approximately 10% of it in 32 km from the dam if compare with biomass in 20-40 m layer of reservoir, the portion of dead increases to 11%. The biomass of zooplankton suspended in the water column of the tail-water sometimes increases (till > 1 g/m3) due to large Copepoda Heteroscope borealis, which inhabits near-bottom and near-shore river zones and can be found in the central part of the river during reproductive period. Limnetic zooplankton from the reservoir cannot be considered as important food for planktivores in the tail-water.


Lignin-Containing Photoactive Resins for 3D Printing by Stereolithography.
Generating compatible and competitive materials that are environmentally sustainable and economically viable is paramount for the success of additive manufacturing using renewable materials. We report the successful application of renewable, modified lignin-containing photopolymer resins in a commercial stereolithography system. Resins were fabricated within operable ranges for viscosity and cure properties, using up to 15% modified lignin by weight. A 4-fold increase in ductility in cured parts with higher lignin concentration is noted compared to commercial stereolithography resins. Excellent print quality was seen in modified lignin resins, with good layer fusion, high surface definition, and visual clarity. These materials can be used to generate new products for additive manufacturing applications and help fill vacant material property spaces, where ductility, sustainability, and application costs are critical.


Exploring by whole exome sequencing patients with initial diagnosis of Rubinstein-Taybi syndrome: the interconnections of epigenetic machinery disorders.
Rubinstein-Taybi syndrome (RSTS) is an autosomal-dominant neurodevelopmental disease affecting 1:125,000 newborns characterized by intellectual disability, growth retardation, facial dysmorphisms and skeletal abnormalities. RSTS is caused by mutations in genes encoding for writers of the epigenetic machinery: CREBBP (~ 60%) or its homologous EP300 (~ 10%). No causative mutation is identified in up to 30% of patients. We performed whole-exome sequencing (WES) on eight RSTS-like individuals who had normal high-resolution array CGH testing and were CREBBP- and EP300-mutation -negative, to identify the molecular cause. In four cases, we identified putatively causal variants in three genes (ASXL1, KMT2D and KMT2A) encoding members of the epigenetic machinery known to be associated with the Bohring-Opitz, Kabuki and Wiedemann-Steiner syndromes. Each variant is novel, de novo, fulfills the ACMG criteria and is predicted to result in loss-of-function leading to haploinsufficiency of the epi-gene. In two of the remaining cases, homozygous/compound heterozygous variants in XYLT2 and PLCB4 genes, respectively, associated with spondyloocular and auriculocondylar 2 syndromes and in the latter an additional candidate variant in XRN2, a gene yet unrelated to any disease, were detected, but their pathogenicity remains uncertain. These results underscore the broad clinical spectrum of Mendelian disorders of the epigenetic apparatus and the high rate of WES disclosure of the genetic basis in cases which may pose a challenge for phenotype encompassing distinct syndromes. The overlapping features of distinct intellectual disability syndromes reflect common pathogenic molecular mechanisms affecting the complex regulation of balance between open and closed chromatin.


Official invitation letters to promote male partner attendance and couple voluntary HIV counselling and testing in antenatal care: an implementation study in Mbeya Region, Tanzania.
The benefits of male partner involvement in antenatal care (ANC) and prevention of mother-to-child transmission of HIV (PMTCT) for maternal and infant health outcomes have been well recognised. However, in many sub-Saharan African settings, male involvement in these services remains low. Previous research has suggested written invitation letters as a way to promote male partner involvement. In this implementation study conducted at three study sites in southwest Tanzania, acceptability of written invitation letters for male partners was assessed. Pre-study CVCT rates of 2-19 % had been recorded at the study sites. Pregnant women approaching ANC without a male partner were given an official letter, inviting the partner to attend a joint ANC and couple voluntary counselling and testing (CVCT) session. Partner attendance was recorded at subsequent antenatal visits, and the invitation was repeated if the partner did not attend. Analysis of socio-demographic indices associated with male partner attendance at ANC was also performed. Out of 318 women who received an invitation letter for their partner, 53.5 % returned with their partners for a joint ANC session; of these, 81 % proceeded to CVCT. Self-reported HIV-positive status at baseline was negatively associated with partner return (p = 0.033). Male attendance varied significantly between the rural and urban study sites (p < 0.001) with rates as high as 76 % at the rural site compared to 31 % at the urban health centre. The majority of women assessed the joint ANC session as a favourable experience, however 7 (75 %) of women in HIV-positive discordant or concordant relationships reported problems during mutual disclosure. Beneficial outcomes reported one month after the session included improved client- provider relationship, improved intra-couple communication and enhanced sexual and reproductive health decision-making. Official invitation letters are a feasible intervention in a resource limited sub-Saharan African context, they are highly accepted by couple members, and are an effective way to encourage men to attend ANC and CVCT. Pre-intervention CVCT rates were improved in all sites. However, urban settings might require extra emphasis to reach high rates of partner attendance compared to smaller rural health centres.


Crystal Structures of the Nuclear Receptor, Liver Receptor Homolog 1, Bound to Synthetic Agonists.
Liver receptor homolog 1 (NR5A2, LRH-1) is an orphan nuclear hormone receptor that regulates diverse biological processes, including metabolism, proliferation, and the resolution of endoplasmic reticulum stress. Although preclinical and cellular studies demonstrate that LRH-1 has great potential as a therapeutic target for metabolic diseases and cancer, development of LRH-1 modulators has been difficult. Recently, systematic modifications to one of the few known chemical scaffolds capable of activating LRH-1 failed to improve efficacy substantially. Moreover, mechanisms through which LRH-1 is activated by synthetic ligands are entirely unknown. Here, we use x-ray crystallography and other structural methods to explore conformational changes and receptor-ligand interactions associated with LRH-1 activation by a set of related agonists. Unlike phospholipid LRH-1 ligands, these agonists bind deep in the pocket and do not interact with residues near the mouth nor do they expand the pocket like phospholipids. Unexpectedly, two closely related agonists with similar efficacies (GSK8470 and RJW100) exhibit completely different binding modes. The dramatic repositioning is influenced by a differential ability to establish stable face-to-face π-π-stacking with the LRH-1 residue His-390, as well as by a novel polar interaction mediated by the RJW100 hydroxyl group. The differing binding modes result in distinct mechanisms of action for the two agonists. Finally, we identify a network of conserved water molecules near the ligand-binding site that are important for activation by both agonists. This work reveals a previously unappreciated complexity associated with LRH-1 agonist development and offers insights into rational design strategies.


Small noncoding RNAs controlling pathogenesis.
Infectious diseases are a leading cause of mortality worldwide. A major challenge in achieving their eradication is a better understanding of bacterial pathogenesis processes. The recent discovery of small noncoding RNAs (sRNAs) as modulators of gene expression in response to environmental cues has brought a new insight into bacterial regulation. sRNAs coordinate complex networks of stress adaptation and virulence gene expression. sRNAs generally ensure such a regulation by pairing to mRNAs of effector and/or regulatory genes, or by binding to proteins. An updated view on bacterial models responsible for important infections illustrates the key role of sRNAs in the control of pathogenesis.


Effect of Foliarly Applied Spirotetramat on Reproduction of Heterodera avenae on Wheat Roots.
The cereal cyst nematode, Heterodera avenae, has the potential to reduce yields of cereal crops in the Pacific Northwest. Spirotetramat (Movento) is a foliar-applied insecticide with ambimobile translocation that reduces fecundity of sucking insects which feed on roots as well as foliage. Spirotetramat (88 g/ha) was applied to foliage during 2010 in two wheat fields infested by H. avenae near St. Anthony, ID and Palouse, WA. In Idaho, two applications at 2-week intervals during late spring to plants already exhibiting swollen white females reduced the postharvest density of H. avenae eggs plus juveniles by 35% (P = 0.03) compared to the nontreated control. In Washington, a single application before white females became apparent reduced the nematode density by 78% (P = 0.01). Grain yields and test weights were not significantly affected by application of spirotetramat at either location. In addition, symptomatic plants from the Idaho field were transplanted into greenhouse pots and treated with spirotetramat. One application (110 g/ha) reduced numbers of eggs plus juveniles/plant by 78% (P = 0.02). Spirotetramat effectively reduced H. avenae populations and warrants further evaluation as a substitute for crop rotations or long fallow periods that reduce nematode population densities in infested fields.


Vitamin D supplementation in pregnancy: a systematic review.
It is unclear whether or not the current evidence base allows definite conclusions to be made regarding the optimal maternal circulating concentration of 25-hydroxyvitamin D [25(OH)D] during pregnancy, and how this might best be achieved. To answer the following questions: (1) What are the clinical criteria for vitamin D deficiency in pregnant women? (2) What adverse maternal and neonatal health outcomes are associated with low maternal circulating 25(OH)D? (3) Does maternal supplementation with vitamin D in pregnancy lead to an improvement in these outcomes (including assessment of compliance and effectiveness)? (4) What is the optimal type (D2 or D3), dose, regimen and route for vitamin D supplementation in pregnancy? (5) Is supplementation with vitamin D in pregnancy likely to be cost-effective? We performed a systematic review and where possible combined study results using meta-analysis to estimate the combined effect size. Major electronic databases [including Database of Abstracts of Reviews of Effects (DARE), Centre for Reviews and Dissemination (CRD), Cochrane Database of Systematic Reviews (CDSR) and the Health Technology Assessment (HTA) database] were searched from inception up to June 2012 covering both published and grey literature. Bibliographies of selected papers were hand-searched for additional references. Relevant authors were contacted for any unpublished findings and additional data if necessary. Abstracts were reviewed by two reviewers. pregnant women or pregnant women and their offspring. either assessment of vitamin D status [dietary intake, sunlight exposure, circulating 25(OH)D concentration] or supplementation of participants with vitamin D or food containing vitamin D (e.g. oily fish). offspring - birthweight, birth length, head circumference, bone mass, anthropometry and body composition, risk of asthma and atopy, small for gestational dates, preterm birth, type 1 diabetes mellitus, low birthweight, serum calcium concentration, blood pressure and rickets; mother - pre-eclampsia, gestational diabetes mellitus, risk of caesarean section and bacterial vaginosis. Seventy-six studies were included. There was considerable heterogeneity between the studies and for most outcomes there was conflicting evidence. The evidence base was insufficient to reliably answer question 1 in relation to biochemical or disease outcomes. For questions 2 and 3, modest positive relationships were identified between maternal 25(OH)D and (1) offspring birthweight in meta-analysis of three observational studies using log-transformed 25(OH)D concentrations after adjustment for potential confounding factors [pooled regression coefficient 5.63 g/10% change maternal 25(OH)D, 95% confidence interval (CI) 1.11 to 10.16 g], but not in those four studies using natural units, or across intervention studies; (2) offspring cord blood or postnatal calcium concentrations in a meta-analysis of six intervention studies (all found to be at high risk of bias; mean difference 0.05 mmol/l, 95% CI 0.02 to 0.05 mmol/l); and (3) offspring bone mass in observational studies judged to be of good quality, but which did not permit meta-analysis. The evidence base was insufficient to reliably answer questions 4 and 5. Study methodology varied widely in terms of study design, population used, vitamin D status assessment, exposure measured and outcome definition. The evidence base is currently insufficient to support definite clinical recommendations regarding vitamin D supplementation in pregnancy. Although there is modest evidence to support a relationship between maternal 25(OH)D status and offspring birthweight, bone mass and serum calcium concentrations, these findings were limited by their observational nature (birthweight, bone mass) or risk of bias and low quality (calcium concentrations). High-quality randomised trials are now required. This study is registered as PROSPERO CRD42011001426. The National Institute for Health Research Health Technology Assessment programme.


Interventions for preventing infectious complications in haemodialysis patients with central venous catheters.
Central venous catheters (CVC) continue to play a prominent role in haemodialysis vascular access with 46% to 70% of patients commencing haemodialysis via a CVC. CVC access is associated with catheter-related infections, increased patient hospitalisations and death due to infection. A variety of interventions are used to prevent CVC infection. To evaluate the benefits and harms of prophylactic topical antimicrobials, topical antiseptics, medicated and non-medicated dressings on infectious complications among haemodialysis patients with CVC. We searched the Cochrane Renal Group's specialised register, the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, EMBASE and reference lists of articles without language restriction. We included randomised controlled trials (RCTs) and quasi-RCTs investigating any intervention that prevented infectious complications among haemodialysis patients with CVC. We excluded antimicrobial impregnated CVC or CVC using locking solutions with antimicrobial properties. Two authors assessed study quality and extracted data. Dichotomous outcomes were expressed as risk ratios (RR) with 95% confidence intervals (CI) and continuous outcomes as mean differences (MD). Ten studies (786 patients) were included. Mupirocin ointment reduced the risk of catheter-related bacteraemia (RR 0.17, 95%CI 0.07 to 0.43) and had a significant effect on catheter-related infections caused by S. aureus. The risk of catheter-related bacteraemia was reduced by polysporin (RR 0.40, 95%CI 0.19 to 0.86) and povidone-iodine ointment (RR 0.10, 95%CI 0.01 to 0.72). Subgroup analysis suggested mupirocin (RR 0.12, 95%CI 0.01 to 2.13) and povidone-iodine ointment (RR 0.84, 95%CI 0.24 to 2.98) had no effect on all-cause mortality while polysporin ointment showed a significant reduction (RR 0.22, 95%CI 0.07 to 0.74). Mortality related to infection was not reduced by mupirocin, polysporin or povidone-iodine ointment. Topical honey did not reduce the risk of exit site infection (RR 0.45, 95%CI 0.10 to 2.11) or catheter-related bacteraemia (RR 0.80, 95%CI 0.37 to 1.73). Transparent polyurethane dressing compared to dry gauze dressing did not reduce the risk of CVC or exit site infection, or catheter-related bacteraemia. Mupirocin ointment appears effective in reducing the risk of catheter-related bacteraemia. Insufficient reporting on mupirocin resistance was noted and needs to be considered in future studies. A lack of high quality data on the routine use of povidone-iodine ointment, polysporin ointment and topical honey warrant larger RCTs. Insufficient data were available to determine which dressing type (transparent polyurethane or dry gauze dressing) has the lowest risk of catheter-related infections.


Crybb2 coding for βB2-crystallin affects sensorimotor gating and hippocampal function.
βB2-crystallin (gene symbol: Crybb2/CRYBB2) was first described as a structural protein of the ocular lens. This gene, however, is also expressed in several regions of the mammalian brain, although its function in this organ remains entirely unknown. To unravel some aspects of its function in the brain, we combined behavioral, neuroanatomical, and physiological analyses in a novel Crybb2 mouse mutant, O377. Behavioral tests with male O377 mutants revealed altered sensorimotor gating, suggesting modified neuronal functions. Since these mouse mutants also displayed reduced hippocampal size, we concentrated further investigations on the hippocampus. Free intracellular Ca(2+) levels were increased and apoptosis was enhanced in the hippocampus of O377 mutants. Moreover, the expression of the gene encoding calpain 3 (gene symbol Capn3) was elevated and the expression of genes coding for the NMDA receptor subunits was downregulated. Additionally, the number of parvalbumin-positive interneurons was decreased in the hippocampus but not in the cortex of the mutants. High-speed voltage-sensitive dye imaging demonstrated an increased translation of input-to-output neuronal activity in the dentate gyrus of this Crybb2 mutant. These results point to an important function of βB2-crystallin in the hippocampal network. They indicate pleiotropic effects of mutations in the Crybb2 gene, which previously had been considered to be specific to the ocular lens. Moreover, our results are the first to demonstrate that βB2-crystallin has a role in hippocampal function and behavioral phenotypes. This model can now be further explored by future experiments.


Alcohol intake and risk of incident psoriasis in US women: a prospective study.
To evaluate the independent association between alcohol consumption and risk of developing psoriasis and to determine if this risk is associated with different types of alcoholic beverages. A prospective study of female nurses who were followed up from 1991 to 2005. Nurses' Health Study II, a cohort of 116,671 US women aged 27 to 44 years in 1991. The study population included 82,869 women who reported amount and type of alcohol intake on biennial questionnaires. We excluded participants with a history of psoriasis prior to 1991. Self-report of incident physician-diagnosed psoriasis. For a sensitivity analysis, we had a subset of confirmed psoriasis cases. There were 1150 cases of incident psoriasis, 1069 of which were used for analysis. Compared with women who did not drink alcohol, the multivariate relative risk (RR) of psoriasis was 1.72 (95% confidence interval [CI], 1.15-2.57) for an alcohol consumption of 2.3 drinks/wk or more. When examined by type of alcoholic beverage, there was an association between psoriasis and nonlight beer intake (multivariate RR for ≥ 5 drinks/wk, 1.76; 95% CI, 1.15-2.69); light beer, red wine, white wine, and liquor were not significantly associated with psoriasis risk. The association with nonlight beer intake became stronger in a subset of confirmed psoriasis cases (multivariate RR for ≥ 5 drinks/wk, 2.29; 95% CI, 1.36-3.85). Nonlight beer intake is associated with an increased risk of developing psoriasis among women. Other alcoholic beverages did not increase the risk of psoriasis in this study.


Generation of candidate ligands for nicotinic acetylcholine receptors via in situ click chemistry with a soluble acetylcholine binding protein template.
Nicotinic acetylcholine receptors (nAChRs), which are responsible for mediating key physiological functions, are ubiquitous in the central and peripheral nervous systems. As members of the Cys loop ligand-gated ion channel family, neuronal nAChRs are pentameric, composed of various permutations of α (α2 to α10) and β (β2 to β4) subunits forming functional heteromeric or homomeric receptors. Diversity in nAChR subunit composition complicates the development of selective ligands for specific subtypes, since the five binding sites reside at the subunit interfaces. The acetylcholine binding protein (AChBP), a soluble extracellular domain homologue secreted by mollusks, serves as a general structural surrogate for the nAChRs. In this work, homomeric AChBPs from Lymnaea and Aplysia snails were used as in situ templates for the generation of novel and potent ligands that selectively bind to these proteins. The cycloaddition reaction between building-block azides and alkynes to form stable 1,2,3-triazoles was used to generate the leads. The extent of triazole formation on the AChBP template correlated with the affinity of the triazole product for the nicotinic ligand binding site. Instead of the in situ protein-templated azide-alkyne cycloaddition reaction occurring at a localized, sequestered enzyme active center as previously shown, we demonstrate that the in situ reaction can take place at the subunit interfaces of an oligomeric protein and can thus be used as a tool for identifying novel candidate nAChR ligands. The crystal structure of one of the in situ-formed triazole-AChBP complexes shows binding poses and molecular determinants of interactions predicted from structures of known agonists and antagonists. Hence, the click chemistry approach with an in situ template of a receptor provides a novel synthetic avenue for generating candidate agonists and antagonists for ligand-gated ion channels.


Amoxapine versus amitriptyline for continuation therapy of depression.
The efficacy of continuation therapy with tricyclic antidepressants has been established in a number of controlled trials. This study investigated the efficacy of continuation therapy with a relatively new antidepressant, amoxapine, using a double-blind controlled comparison with amitriptyline. Subjects met DSM-III criteria for major depressive disorder and were randomized to treatment with either amoxapine 400 mg (N = 47) or amitriptyline 300 mg (N = 45). The acute phase lasted up to 8 weeks. Responders were continued on the same drug at the same dose for a 16-week continuation phase. Some measures found more rapid onset for amitriptyline, which is inconsistent with findings from some prior studies. Amitriptyline was more effective in inducing full recovery. There was a trend for higher relapse rates on amoxapine, perhaps related to the fact that there were more partial responders entering continuation therapy from this group. Side effect rates were equivalent in the two drugs. However, physicians rated amoxapine's side effects as more frequently interfering with its therapeutic effect. These data suggest that amoxapine does not offer any clear advantage over amitriptyline for continuation therapy in patients who have major depressive disorder. Of potential clinical relevance is the finding that achieving full recovery in the acute phase may reduce the likelihood of relapse in the continuation phase, regardless of the type of antidepressant medication prescribed.


Safety and efficacy of cold atmospheric plasma for the sterilization of a Pasteurella multocida-contaminated subcutaneously implanted foreign body in rabbits.
To determine whether a stainless steel implant sterilized with a novel cold atmospheric plasma sterilization (CAPS) device adversely affects local tissues in rabbits and whether CAPS was as effective as steam sterilization with an autoclave to inactivate Pasteurella multocida. 31 healthy New Zealand White rabbits. Steam-autoclaved stainless steel implants inoculated with P multocida underwent a second steam autoclave sterilization (AIA) or CAPS (AICAPS). One AIA implant and 3 AICAPS implants were randomly placed subcutaneously at 4 sites in 21 rabbits (84 implants). These rabbits were monitored daily for 5 days for evidence of systemic illness and local tissue reactions at the implantation sites and then euthanized. Samples were taken from each implant site for bacterial culture and histologic examination. Cultures of samples obtained from all sites were negative for bacterial growth. No significant difference was observed in mean skin thickness or erythema between AIA and AICAPS implant sites on any observed day. Also, individual histologic grades for the epidermis, dermis, subcutis, and muscle and total histologic grade were not significantly different between AIA and AICAPS implant sites. Cold atmospheric plasma sterilization was noninferior to steam sterilization of P multocida-contaminated stainless steel implants in the rabbits in the present study. However, studies of the efficacy of CAPS for inactivation of other important bacteria are needed.


Phase II/III weekly nab-paclitaxel plus gemcitabine or carboplatin versus gemcitabine/carboplatin as first-line treatment of patients with metastatic triple-negative breast cancer (the tnAcity study): study protocol for a randomized controlled trial.
Triple-negative breast cancer is an aggressive disease with unmet clinical needs. In a phase III study of patients with metastatic triple-negative breast cancer, first-line gemcitabine/carboplatin resulted in a median progression-free survival of 4.6 months. nab-paclitaxel-based regimens (with gemcitabine or carboplatin±bevacizumab) also demonstrated efficacy and safety in first-line phase II trials of human epidermal growth factor receptor 2-negative metastatic breast cancer. In this international, multicenter, open-label, randomized phase II/III trial, the efficacy and safety of first-line nab-paclitaxel with gemcitabine or with carboplatin will be compared with gemcitabine/carboplatin (control arm) for metastatic triple-negative breast cancer. In the phase II portion, 240 patients with measurable metastatic triple-negative breast cancer and treatment-naive for metastatic disease will be randomized 1:1:1 (stratified by disease-free interval: ≤1 versus>1 year) to nab-paclitaxel 125 mg/m2 plus gemcitabine 1000 mg/m2, nab-paclitaxel 125 mg/m2 plus carboplatin area under the curve 2 mg×min/mL, or gemcitabine 1000 mg/m2 plus carboplatin area under the curve 2 mg×min/mL, all given on days 1 and 8 of a 21-day cycle. Investigator-assessed progression-free survival (primary endpoint), overall response rate, overall survival, and safety will be assessed. A ranking algorithm of five efficacy and safety parameters will be used to pick the "winner" of the nab-paclitaxel regimens. In the phase III portion, 550 patients will be randomized 1:1 (stratified by disease-free interval: ≤1 versus >1 year, and prior adjuvant/neoadjuvant taxane use) to the nab-paclitaxel combination arm selected from the phase II portion or to the control arm. Patients in phase II will not be part of the phase III population. The phase III primary endpoint is blinded, independently-assessed progression-free survival; secondary endpoints include blinded, independently-assessed overall response rate, overall survival, disease control rate, duration of response, and safety. Biomarker and circulating tumor-cell exploratory analyses and quality-of-life assessments will also be performed. A list of approving ethical bodies was provided in Additional file 1. The tnAcity trial aims to identify a new standard cytotoxic chemotherapy regimen for first-line treatment of metastatic triple-negative breast cancer. ClinicalTrials.gov: NCT01881230 . Date of registration: 17 June 2013.


Performance and inter-observer variability of prostate MRI (PI-RADS version 2) outside high-volume centres.
Objective: Despite the growing trend to embrace pre-biopsy MRI in the diagnostic pathway for prostate cancer (PC), its performance and inter-observer variability outside high-volume centres remains unknown. This study aims to evaluate sensitivity of and variability between readers of prostate MRI outside specialized units with radical prostatectomy (RP) specimen as the reference standard.Materials and methods: Retrospective study comprising a consecutive cohort of all 97 men who underwent MRI and subsequent RP between January 2012 and December 2014 at a private hospital in Sweden. Three readers, blinded to clinical data, reviewed all images (including 11 extra prostate MRI to reduce bias). A tumour was considered detected if the overall PI-RADS v2 score was 3-5 and there was an approximate match (same or neighbouring sector) of tumour sector according to a 24 sector system used for both MRI and whole mount sections.Results: Detection rate for the index tumour ranged from 67 to 76%, if PI-RADS 3-5 lesions were considered positive and 54-66% if only PI-RADS score 4-5 tumours were included. Detection rate for aggressive tumours (GS ≥ 4 + 3) was higher; 83.1% for PI-RADS 3-5 and 79.2% for PI-RADS 4-5. The agreement between readers showed average [Formula: see text] values of 0.41 for PI-RADS score 3-5 and 0.51 for PI-RADS score 4-5.Conclusions: Prostate MRI evidenced a moderate detection rate for clinically significant PC with a rather large variability between readers. Clinics outside specialized units must have knowledge of their performance of prostate MRI before considering omitting biopsies in men with negative MRI.


Hyperbaric oxygen as a chemotherapy adjuvant in the treatment of osteosarcoma.
Although hyperbaric oxygen has been shown to enhance the efficacy of radiotherapy and chemotherapy for the treatment of several malignant tumors, the impact of hyperbaric oxygen on osteosarcoma has not yet been demonstrated. In this study, we investigated the efficacy of hyperbaric oxygen alone and in combination with an anti-cancer drug as an adjuvant to chemotherapy. In vitro, highly metastatic murine osteosarcoma cell lines were exposed to hyperbaric oxygen and cell viability was examined. Hyperbaric oxygen alone significantly suppressed cell proliferation, and hyperbaric oxygen plus carboplatin exhibited significant synergism in suppression of cell proliferation. In vivo, C3H mice were subcutaneously inoculated with osteosarcoma cells and divided into four groups: control, hyperbaric oxygen, carboplatin, and carboplatin plus hyperbaric oxygen. After 5 weeks, increase in both tumor volume and number of lung metastases was significantly suppressed in the hyperbaric oxygen group. Concomitant hyperbaric oxygen clearly enhanced the chemotherapeutic effects of carboplatin on both tumor growth and lung metastasis in osteosarcoma-bearing mice. Moreover, mortality in the carboplatin plus hyperbaric oxygen group was significantly lower than in the other three groups. These findings suggest that hyperbaric oxygen plus carboplatin combination therapy could be an appropriate therapeutic regimen for the treatment of patients with osteosarcoma.


Multiplexed shotgun genotyping for rapid and efficient genetic mapping.
We present a new approach to genotyping based on multiplexed shotgun sequencing that can identify recombination breakpoints in a large number of individuals simultaneously at a resolution sufficient for most mapping purposes, such as quantitative trait locus (QTL) mapping and mapping of induced mutations. We first describe a simple library construction protocol that uses just 10 ng of genomic DNA per individual and makes the approach accessible to any laboratory with standard molecular biology equipment. Sequencing this library results in a large number of sequence reads widely distributed across the genomes of multiplexed bar-coded individuals. We develop a Hidden Markov Model to estimate ancestry at all genomic locations in all individuals using these data. We demonstrate the utility of the approach by mapping a dominant marker allele in D. simulans to within 105 kb of its true position using 96 F1-backcross individuals genotyped in a single lane on an Illumina Genome Analyzer. We further demonstrate the utility of our method by genetically mapping more than 400 previously unassembled D. simulans contigs to linkage groups and by evaluating the quality of targeted introgression lines. At this level of multiplexing and divergence between strains, our method allows estimation of recombination breakpoints to a median of 38-kb intervals. Our analysis suggests that higher levels of multiplexing and/or use of strains with lower levels of divergence are practicable.


Effects of Corrective Exercise for Thoracic Hyperkyphosis on Posture, Balance, and Well-Being in Older Women: A Double-Blind, Group-Matched Design.
The purpose of this study was to identify the effects of a corrective exercise for thoracic hyperkyphosis on posture, balance, and well-being in Korean community-dwelling older women. Fifty women 65 years of age and older, recruited from 2 senior centers, participated in this study. Participants were assigned to either the experimental group (EG) or the control group (CG) on the basis of convenience of location, and 22 in each were analyzed. Participants in the EG underwent a thoracic corrective exercise program 1 hour each session, twice per week for 8 weeks (a total of 16 sessions), which consisted of specific exercises to enhance breathing, thoracic mobility and stability, and awareness of thoracic alignment. The CG received education on the same thoracic corrective exercise program and a booklet of the exercises. Outcome measures included the extent of postural abnormality (angle of thoracic kyphosis, kyphosis index calculated both in relaxed- and best posture using flexicurve, the ratio of the kyphosis index calculated best posture/relaxed posture, craniovertebral angle, and tragus-to-wall distance), balance (Short Physical Performance Battery and limit of stability), and well-being (Geriatric Depression Scale Short Form and the 36-Item Short Form Health Survey [SF-36]). All data were collected by 6 blinded assessors at baseline, at 8 weeks after the completion of intervention, and at 16 weeks for follow-up. For participants of the EG, means of all parameters showed significant improvements over time (P < .05), with improved values both in comparison of baseline to postintervention and baseline to follow-up. Means of CG parameters were significantly improved in only the angle of thoracic kyphosis and the tragus-to-wall distance (P < .05). Furthermore, in all parameters, percent change between baseline and postintervention data was significantly (P < .05) higher for the EG than that for the CG, except for the limit of stability and SF-36 which improved but not significantly. All parameters between baseline and follow-up data were significantly (P < .05) higher for the EG than those for the CG, except for the limit of stability. The findings of this study suggest that a well-designed exercise program may be beneficial to improve spinal posture, balance, and well-being in older women with thoracic hyperkyphosis. We recommend the use of the therapeutic strategies utilized in this study to enhance thoracic posture, balance, and well-being of older women with thoracic hyperkyphosis. Future research is needed to apply this exercise protocol on a larger and more diverse population.


Cardiac myocytes control release of endothelin-1 in coronary vasculature.
Alpha-adrenergic vasoconstriction in the coronary circulation is mediated through alpha-adrenoceptors on cardiac myocytes and subsequent release of endothelin, a very potent, long-lasting vasoconstrictor. Recent studies found that adult cardiac myocytes do not express the preproendothelin gene. Thus we hypothesized that alpha-adrenoceptor stimulation on the cardiac myocytes results in the production of an endothelin-releasing factor, which stimulates the coronary vasculature to produce endothelin. We tested this hypothesis by using an in vitro model in which isolated adult rat cardiac myocytes can be stimulated with an alpha-adrenoceptor agonist (phenylephrine). Their bathing fluid is then transferred to isolated coronary arterioles, and vasoactive responses are measured. To identify the source of endothelin, the endothelin-converting enzyme inhibitor phosphoramidon was added to either the myocytes or the isolated arterioles. Phenylephrine enhanced the vasoconstrictor properties of the myocyte bathing fluid. Administration of phosphoramidon (in either the presence or the absence of phenylephrine) to the myocytes had no effect on the vasoactive properties of the bathing fluid. In contrast, administration of phosphoramidon to the isolated arteriole before administration of the bathing fluid converted vasoconstriction to vasodilation, similar to the effect of the endothelin A receptor antagonist JKC-301, indicating that the endothelin is indeed produced by the coronary vasculature. Administration of the angiotensin type 1 receptor antagonist losartan to the vessel bath enhanced vasodilation to the bathing fluid of the phenylephrine-treated but not control myocytes. In conclusion, during alpha-adrenergic activation cardiac myocytes release a factor, probably angiotensin II, that stimulates the vascular production of endothelin. Although the physiological implications of this mechanism are not obvious, this may represent a protective mechanism that integrates neuronal vasoconstrictor mechanisms with myocardial metabolism, which minimizes periods of both coronary underperfusion and overperfusion.


Healthcare Professionals' Knowledge of Pharmacogenetics and Attitudes Towards Antimicrobial Utilization in Zambia: Implications for a Precision Medicine Approach to Reducing Antimicrobial Resistance.
Introduction: Sub-Saharan Africa and other low- and middle-income countries (LMICs) have the highest rates of antimicrobial resistance (AMR) driven by high rates of antimicrobial utilization. This is a concern as AMR appreciably increases morbidity, mortality and costs. Pharmacogenetics (PGx) and precision medicine are emerging approaches to combat AMR. Consequently, as a first step there is a need to assess AMR knowledge and attitudes, and knowledge of PGx, among healthcare professionals and use the findings to guide future interventions. Methodology: We conducted a cross-sectional study involving 304 healthcare professionals at tertiary hospitals in Lusaka, Zambia. Structural Equation Modeling (SEM) was used to analyze relationships among latent variables. Results: Overall correctness of answers concerning AMR among healthcare professionals was 60.4% (7/11). Knowledge of pharmacogenetics was low (38%). SEM showed that high AMR knowledge score correlated with a positive attitude toward combating AMR (p < 0.001). Pharmacists had relatively higher AMR knowledge scores (mean = 7.67, SD = 1.1), whereas nurses had lower scores (mean = 5.57, SD = 1.9). A minority of respondents [31.5% (n = 95)] indicated that poor access to local antibiogram data promoted AMR, with the majority [56.5% (n = 190)] responding that poor adherence to prescribed antimicrobials can lead to AMR. Pharmacists had the highest scores for attitude (mean = 5.60, SD = 1.6) whereas nurses had the lowest scores (mean = 4.02, SD = 1.4). Conclusion: AMR knowledge and attitudes, as well as knowledge on PGx among healthcare professionals in Zambia, is sub-optimal and has the potential to affect the uptake of precision medicine approaches to reduce AMR rates. Educational and positive behavioral change interventions are required to address this and in future, we will be seeking to introduce these to improve the use of antimicrobials.


Enzyme immunosorbent assay for Ebola virus antigens in tissues of infected primates.
A sandwich enzyme immunosorbent assay (EIA) using a mixture of mouse monoclonal antibodies for antigen capture and polyclonal hyperimmune rabbit anti-Ebola virus serum for antigen detection was developed and evaluated on the tissues of monkeys naturally or experimentally infected with strains of Ebola viruses. When compared with virus isolation, the antigen detection EIA was both sensitive and specific: 44 of 45 (97.7%) liver homogenates and 38 of 41 (92.7%) spleen homogenates that were culture positive and tested by both techniques were positive for viral antigen, while 85 of 87 (97.7%) culture-negative liver homogenates and 66 of 66 culture-negative spleen homogenates were found to be antigen negative. The assay, initially developed to detect antigens of prototype African strains of Ebola virus, reliably detected related strains of Ebola virus found during two recent outbreaks of Ebola virus infection among imported, quarantined Macaca fascicularis monkeys in the United States. The assay allows economical and rapid testing of large numbers of tissue specimens. Antigen was found in homogenates of spleen and liver and in serum.


To What Extent Does the EU General Data Protection Regulation (GDPR) Apply to Citizen Scientist-Led Health Research with Mobile Devices?
In this article, we consider the possible application of the European General Data Protection Regulation (GDPR) to "citizen scientist"-led health research with mobile devices. We argue that the GDPR likely does cover this activity, depending on the specific context and the territorial scope. Remaining open questions that result from our analysis lead us to call for lex specialis that would provide greater clarity and certainty regarding the processing of health data by for research purposes, including these non-traditional researchers.


The use of the direct linear plot for determining initial velocities.
A new method is described for estimating initial velocities of enzyme-catalysed reactions. It is simple to apply either graphically or numerically, and is particularly appropriate for experiments in which the initial straight part of the progress curve is very short or non-existent. It requires no more knowledge than is readily available about the details of the system, such as the extent of reaction at equilibrium, the rate of enzyme inactivation, the nature of product inhibition etc., unlike some other methods of analysing progress curves, which are often invalidated by small errors in the defining assumptions.


Homologous pairs of regulatory proteins control activity of Bacillus subtilis transcription factor sigma(b) in response to environmental stress.
In Bacillus subtilis, activity of the general stress transcription factor sigma B is controlled posttranslationally by a regulatory network that transmits signals of environmental and metabolic stress. These signals include heat, ethanol, or osmotic challenge, or a sharp decrease in cellular energy levels, and all ultimately control sigma B activity by influencing the binding decision of the RsbW anti-sigma factor. In the absence of stress, RsbW binds to sigma B and prevents its association with RNA polymerase core enzyme. However, following stress, RsbW binds instead to the RsbV anti-anti-sigma factor, thereby releasing sigma B to direct transcription of its target genes. These two principal regulators of sigmaB activity are encoded in the eight-gene sigB operon, which has the gene order rsbR-rsbS-rsbT-rsbU-rsbV-rsbW-sig B-rsbX (where rsb stands for regulator of sigma B). Notably, the predicted rsbS product has significant amino acid identity to the RsbV anti-anti-sigma factor and the predicted rsbT product resembles the RsbW anti-sigma factor. To determine the roles of rsbS and rsbT, null or missense mutations were constructed in the chromosomal copies or each and tested for their effects on expression of a sigma B-dependent reporter fusion. On the basis of this genetic analysis, our principal conclusions are that (i) the rsbS product is a negative regulator of or" activity, (ii) the rsbT product is a positive regulator, (iii) RsbS requires RsbT for function, and (iv) the RsbS-RsbT and RsbV-RsbW pairs act hierarchically by a common mechanism in which key protein-protein interactions are controlled by phosphorylation events.


A Pyrazolo[3,4-d]pyrimidin-4-amine Derivative Containing an Isoxazole Moiety Is a Selective and Potent Inhibitor of RET Gatekeeper Mutants.
Aberrant RET kinase signaling plays critical roles in several human cancers such as thyroid carcinoma. The gatekeeper mutants (V804L or V804M) of RET are resistant to currently approved RET inhibitors such as cabozantinib and vandetanib. We, for the first time, report a highly selective and extremely potent RET inhibitor, 6i rationally designed. Compound 6i inhibits strongly RET gatekeeper mutants and other clinically relevant RET mutants as well as wt-RET. This substance also significantly suppresses growth of thyroid cancer-derived TT cell lines and Ba/F3 cells transformed with various RET mutants. Docking studies reveal that the isoxazole moiety in 6i is responsible for binding affinity improvement by providing additional site for H-bonding with Lys758. Also, 6i not only substantially blocks cellular RET autophosphorylation and its downstream pathway, it markedly induces apoptosis and anchorage-independent growth inhibition in TT cell lines while having no effect on normal thyroid Nthy ori-3-1 cells.


Macrophage-inducible C-type lectin Mincle-expressing dendritic cells contribute to control of splenic Mycobacterium bovis BCG infection in mice.
The macrophage-inducible C-type lectin Mincle has recently been identified to be a pattern recognition receptor sensing mycobacterial infection via recognition of the mycobacterial cell wall component trehalose-6',6-dimycolate (TDM). However, its role in systemic mycobacterial infections has not been examined so far. Mincle-knockout (KO) mice were infected intravenously with Mycobacterium bovis BCG to mimic the systemic spread of mycobacteria under defined experimental conditions. After intravenous infection with M. bovis BCG, Mincle-KO mice responded with significantly higher numbers of mycobacterial CFU in spleen and liver, while reduced granuloma formation was observed only in the spleen. At the same time, reduced Th1 cytokine production and decreased numbers of gamma interferon-producing T cells were observed in the spleens of Mincle-KO mice relative to the numbers in the spleens of wild-type (WT) mice. The effect of adoptive transfer of defined WT leukocyte subsets generated from bone marrow cells of zDC(+/DTR) mice (which bear the human diphtheria toxin receptor [DTR] under the control of the classical dendritic cell-specific zinc finger transcription factor zDC) to specifically deplete Mincle-expressing classical dendritic cells (cDCs) but not macrophages after diphtheria toxin application on the numbers of splenic and hepatic CFU and T cell subsets was then determined. Adoptive transfer experiments revealed that Mincle-expressing splenic cDCs rather than Mincle-expressing macrophages contributed to the reconstitution of attenuated splenic antimycobacterial immune responses in Mincle-KO mice after intravenous challenge with BCG. Collectively, we show that expression of Mincle, particularly by cDCs, contributes to the control of splenic M. bovis BCG infection in mice.


Knowledge Level, Attitude, and Behaviors of Farmers in Çukurova Region regarding the Use of Pesticides.
Farmers are particularly at high risk of pesticide exposure due to added risk from occupational exposure. The aim of this study is to evaluate knowledge level of farmers in the Çukurova region of the effects of pesticides, toxic symptoms, and protective equipment as well as assessing their attitudes and practices regarding pesticides. A total of 420 seasonal agricultural workers in Karataş District of Adana Province, Turkey, were included in the study. The questionnaire form consisting of 3 sections was administered using face-to-face interview method. The mean age of the participants was 40.2 ± 10.6 years. They were engaged in farming for a mean duration of 18.5 ± 10.6 years. All of them used pesticides, but none of them had been trained on the use of pesticides. Only 26.2% of them stored pesticides in a private depot. The farmers who took empty pesticide containers to special collection bins or centers constituted only 4.3% of them. While 84.0% of them thought that pesticides could have a negative impact on human health, 5.0% of them had experienced a medical problem and 1.0% of them were poisoned after applying pesticides. The results show that knowledge level of farmers of safe use of pesticides is very inadequate. This lack of knowledge adversely affects workers' quality of life as well as occupational health and safety. Appropriate training programs should be organized to increase their level of knowledge.