Practice patterns for the elicitation of sexual history, education, and counseling among providers of STD services: results from the gonorrhea community action project (GCAP).
The frequently asymptomatic nature and high incidence of severe complications of sexually transmitted diseases (STD) calls for targeted efforts to identify those at greatest risk. Earlier studies have shown inconsistencies regarding STD evaluation by primary care clinicians and physicians. However, the literature regarding the consistency of practice patterns regarding elicitation of sexual history is limited. We examined practice patterns for the elicitation of sexual history among providers across seven sites nationwide. As part of a multisite study to encourage health seeking for populations specifically at risk for gonorrhea (GC) and other STDs, semistructured interviews that included questions regarding sexual history elicitation were conducted with 208 service providers in a total of 121 publicly and privately funded clinics, managed care organizations (MCOs), hospital clinics, community- and school-based clinics in Denver, New York, Los Angeles, Birmingham, St. Louis, Indianapolis, and Prince Georges County, MD. Among the providers interviewed, practice patterns for the elicitation of sexual history were inconsistent. Sexual histories were described as routine (i.e., solicited from every client regardless of reason for visit) in 57% of sites. Providers most frequently asked clients their number of sex partners (57%), their contraceptive history (55%), and STD history (34%). Client discomfort among 46% and provider discomfort among 13% was cited as barriers to the elicitation of sexual history. A quarter (26%) of providers agreed that the elicitation of sexual history can be fostered by improved provider communication skills and 16% agreed increasing training and experience for providers is needed. These findings suggest that interventions with providers to standardize sexual history elicitation can help to reduce barriers to prevention, diagnosis, and treatment of STD.


Pre-validation methods for developing a patient reported outcome instrument.
Measures that reflect patients' assessment of their health are of increasing importance as outcome measures in randomised controlled trials. The methodological approach used in the pre-validation development of new instruments (item generation, item reduction and question formatting) should be robust and transparent. The totality of the content of existing PRO instruments for a specific condition provides a valuable resource (pool of items) that can be utilised to develop new instruments. Such 'top down' approaches are common, but the explicit pre-validation methods are often poorly reported. This paper presents a systematic and generalisable 5-step pre-validation PRO instrument methodology. The method is illustrated using the example of the Aberdeen Glaucoma Questionnaire (AGQ). The five steps are: 1) Generation of a pool of items; 2) Item de-duplication (three phases); 3) Item reduction (two phases); 4) Assessment of the remaining items' content coverage against a pre-existing theoretical framework appropriate to the objectives of the instrument and the target population (e.g. ICF); and 5) qualitative exploration of the target populations' views of the new instrument and the items it contains. The AGQ 'item pool' contained 725 items. Three de-duplication phases resulted in reduction of 91, 225 and 48 items respectively. The item reduction phases discarded 70 items and 208 items respectively. The draft AGQ contained 83 items with good content coverage. The qualitative exploration ('think aloud' study) resulted in removal of a further 15 items and refinement to the wording of others. The resultant draft AGQ contained 68 items. This study presents a novel methodology for developing a PRO instrument, based on three sources: literature reporting what is important to patient; theoretically coherent framework; and patients' experience of completing the instrument. By systematically accounting for all items dropped after the item generation phase, our method ensures that the AGQ is developed in a transparent, replicable manner and is fit for validation. We recommend this method to enhance the likelihood that new PRO instruments will be appropriate to the research context in which they are used, acceptable to research participants and likely to generate valid data.


Correlation between small-volume spinal cord doses for spine stereotactic body radiotherapy (SBRT).
Doses to small spinal cord isodose volume (such as those ranging from Dmax 0.0 cc to 0.5 cc) as well as to large volumes (such as those ranging from 0.5 cc to 3.0 cc) are critical parameters to guide safe practice of spine SBRT. We here report a mathematical formula that links the most probable dose volume limits together for common spine SBRT cases.Methods and materials: A dose ripple formula parameterized with equivalent dose radius (EDR) was derived to model spinal cord small-volume doses for a spine SBRT treatment. A cohort of spine SBRT cases (n=68), treated with either a robotic x-band linac or a conventional S-band linac, was selected to verify the model predictions. The mean prescription dose was 22± 4 Gy (range, 12-40 Gy) delivered in 2±1 fractions. The mean and median target volume was 39.4±42.5 cc and 30.3 cc (range, 0.24-264.2 cc), respectively. Direct correlations between the spinal cord Dmax and variable spinal cord doses of increasing isodose volumes (ranging from 0.0 cc to 3.0 cc) of different planning organ-at-risk volumes (PRVs) were investigated. The PRV structures for the study included the true cord, thecal sac and the true cord plus variable margins ranging from 1.0 mm to 3.0 mm. No direct linear correlation was observed amongst the small volume doses to the spinal cord PRVs. However, strong linear correlations (R2 > 0.96) for all the studied PRVs were observed when correlating EDRs amongst isodose volumes ranging from 0.0 cc to 3.0 cc. In particular, EDR dependence was found to differ significantly for the thecal sac versus the spinal cord with or without 1-3 millimeter margins. With strong EDR correlation, the most probable relationship among the small-volume dose limits was derived for the spinal cord PRVs. An analytical formula linked the most probable pin-point/small isodose volume doses with relatively large isodose volume doses of the spinal cord for spine SBRT. As a result, a small number of dose limits such as Dmax or D(0.35cc) are likely sufficient to surrogate the spinal cord dose tolerance for consistent treatment planning optimization and outcome analysis.


Structure and drug resistance of the Plasmodium falciparum transporter PfCRT.
The emergence and spread of drug-resistant Plasmodium falciparum impedes global efforts to control and eliminate malaria. For decades, treatment of malaria has relied on chloroquine (CQ), a safe and affordable 4-aminoquinoline that was highly effective against intra-erythrocytic asexual blood-stage parasites, until resistance arose in Southeast Asia and South America and spread worldwide1. Clinical resistance to the chemically related current first-line combination drug piperaquine (PPQ) has now emerged regionally, reducing its efficacy2. Resistance to CQ and PPQ has been associated with distinct sets of point mutations in the P. falciparum CQ-resistance transporter PfCRT, a 49-kDa member of the drug/metabolite transporter superfamily that traverses the membrane of the acidic digestive vacuole of the parasite3-9. Here we present the structure, at 3.2 Å resolution, of the PfCRT isoform of CQ-resistant, PPQ-sensitive South American 7G8 parasites, using single-particle cryo-electron microscopy and antigen-binding fragment technology. Mutations that contribute to CQ and PPQ resistance localize primarily to moderately conserved sites on distinct helices that line a central negatively charged cavity, indicating that this cavity is the principal site of interaction with the positively charged CQ and PPQ. Binding and transport studies reveal that the 7G8 isoform binds both drugs with comparable affinities, and that these drugs are mutually competitive. The 7G8 isoform transports CQ in a membrane potential- and pH-dependent manner, consistent with an active efflux mechanism that drives CQ resistance5, but does not transport PPQ. Functional studies on the newly emerging PfCRT F145I and C350R mutations, associated with decreased PPQ susceptibility in Asia and South America, respectively6,9, reveal their ability to mediate PPQ transport in 7G8 variant proteins and to confer resistance in gene-edited parasites. Structural, functional and in silico analyses suggest that distinct mechanistic features mediate the resistance to CQ and PPQ in PfCRT variants. These data provide atomic-level insights into the molecular mechanism of this key mediator of antimalarial treatment failures.


Generation and analysis of novel plant-derived antibody-based therapeutic molecules against West Nile virus.
Previously, our group engineered a plant-derived monoclonal antibody (MAb) (pHu-E16) that efficiently treated West Nile virus (WNV) infection in mice. In this study, we developed several pHu-E16 variants to improve its efficacy. These variants included a single-chain variable fragment (scFv) of pHu-E16 fused to the heavy chain (HC) constant domains (CH(1-3)) of human IgG (pHu-E16scFv-CH(1-3)) and a tetravalent molecule (Tetra pHu-E16) assembled from pHu-E16scFv-CH(1-3) with a second pHu-E16scFv fused to the light chain (LC) constant region. pHu-E16scFv-CH(1-3) and Tetra pHu-E16 were efficiently expressed and assembled in plants. To assess the impact of differences in N-linked glycosylation on pHu-E16 variant assembly and function, we expressed additional pHu-E16 variants with various combinations of HC and LC components. Our study revealed that proper pairing of HC and LC was essential for the complete N-glycan processing of antibodies in both plant and animal cells. Associated with their distinct N-glycoforms, pHu-E16, pHu-E16scFv-CH(1-3) and Tetra pHu-E16 exhibited differential binding to C1q and specific Fcγ receptors (FcγR). Notably, none of the plant-derived Hu-E16 variants showed antibody-dependent enhancement (ADE) activity in CD32A+ human cells, suggesting the potential of plant-produced antibodies to minimize the adverse effect of ADE. Importantly, all plant-derived MAb variants exhibited at least equivalent in vitro neutralization and in vivo protection in mice compared to mammalian cell-produced Hu-E16. This study demonstrates the capacity of plants to express and assemble a large, complex and functional IgG-like tetravalent mAb variant and also provides insight into the relationship between MAb N-glycosylation, FcγR and C1q binding, and ADE. These new insights may allow the development of safer and cost effective MAb-based therapeutics for flaviviruses, and possibly other pathogens.


Water quality and health in northern Canada: stored drinking water and acute gastrointestinal illness in Labrador Inuit.
One of the highest self-reported incidence rates of acute gastrointestinal illness (AGI) in the global peer-reviewed literature occurs in Inuit communities in the Canadian Arctic. This high incidence of illness could be due, in part, to the consumption of contaminated water, as many northern communities face challenges related to the quality of municipal drinking water. Furthermore, many Inuit store drinking water in containers in the home, which could increase the risk of contamination between source and point-of-use (i.e., water recontamination during storage). To examine this risk, this research characterized drinking water collection and storage practices, identified potential risk factors for water contamination between source and point-of-use, and examined possible associations between drinking water contamination and self-reported AGI in the Inuit community of Rigolet, Canada. The study included a cross-sectional census survey that captured data on types of drinking water used, household practices related to drinking water (e.g., how it was collected and stored), physical characteristics of water storage containers, and self-reported AGI. Additionally, water samples were collected from all identified drinking water containers in homes and analyzed for presence of Escherichia coli and total coliforms. Despite municipally treated tap water being available in all homes, 77.6% of households had alternative sources of drinking water stored in containers, and of these containers, 25.2% tested positive for total coliforms. The use of transfer devices and water dippers (i.e., smaller bowls or measuring cups) for the collection and retrieval of water from containers were both significantly associated with increased odds of total coliform presence in stored water (ORtransfer device = 3.4, 95% CI 1.2-11.7; ORdipper = 13.4, 95% CI 3.8-47.1). Twenty-eight-day period prevalence of self-reported AGI during the month before the survey was 17.2% (95% CI 13.0-22.5), which yielded an annual incidence rate of 2.4 cases per person per year (95% CI 1.8-3.1); no water-related risk factors were significantly associated with AGI. Considering the high prevalence of, and risk factors associated with, indicator bacteria in drinking water stored in containers, potential exposure to waterborne pathogens may be minimized through interventions at the household level.


Ruptured hydatid cyst presenting as pneumothorax.
Patients with echinococcus infection are mostly asymptomatic. The documented rates of simple pneumothorax in patients with pulmonary hydatidosis ranged from 2.4% - 6.2%. We report a case of a forty-year-old male patient who was referred to our hospital for management of recurrent pneumothorax. A video assisted thoracoscope (VATS) was first introduced which showed a large amount of pus in the pleural cavity and a perforated hydatid cyst. The VATS was converted to an open thoracotomy and decortication was done with removal of the ruptured hydatid. The patient made an unremarkable recovery and was discharged after one week with empyema tubes. The empyema tubes were gradually removed over a period of six weeks. An extraordinary number of management options for pulmonary hydatid disease have been offered. This case report highlights surgical treatment as the management opti.


Increased formate overflow is a hallmark of oxidative cancer.
Formate overflow coupled to mitochondrial oxidative metabolism\ has been observed in cancer cell lines, but whether that takes place in the tumor microenvironment is not known. Here we report the observation of serine catabolism to formate in normal murine tissues, with a relative rate correlating with serine levels and the tissue oxidative state. Yet, serine catabolism to formate is increased in the transformed tissue of in vivo models of intestinal adenomas and mammary carcinomas. The increased serine catabolism to formate is associated with increased serum formate levels. Finally, we show that inhibition of formate production by genetic interference reduces cancer cell invasion and this phenotype can be rescued by exogenous formate. We conclude that increased formate overflow is a hallmark of oxidative cancers and that high formate levels promote invasion via a yet unknown mechanism.


The contribution of parental smoking history and socio-demographic factors to the smoking behavior of Israeli women.
This study examined the interplay between sociodemographic factors and parental smoking history in shaping the smoking behavior of Israeli women (N = 302). The study was conducted in the Negev region, which is characterized by a high proportion of immigrants and high percentage of low socioeconomic and educational groups. The specific objectives of this study were to examine: (1) The prevalence and characteristics of women smokers, ex-smokers and never-smokers; and (2) the contribution of education and parent smoking history to women's current smoking. Low levels of education, being Israeli born or veteran immigrants of European-American origin significantly increased the risk of smoking, whereas an orthodox lifestyle and new immigrant status significantly reduced the likelihood of smoking. Occasional smokers reported significantly higher primary care utilization than never smokers. A significant relationship between smoking and pain, gynecological symptoms and depression was found. Results indicate that childhood exposure to maternal smoking was a significant risk factor for smoking, whereas paternal past smoking negatively affects smoking in women. Also, results show that parental educational level affects women's smoking behavior indirectly by influencing their own educational attainment, which in turn is negatively associated with the likelihood of smoking. Mothers with higher education were more likely to smoke, an effect that was reversed for their daughters. Our results demonstrate how demographic, parental and lifestyle factors affect women's smoking in a multi-ethnic society and highlight the need to examine both generational and intergenerational effects.


Ethnic differences in socioeconomic status, diagnosis, treatment, and survival among older women with epithelial ovarian cancer.
The purpose of the study was to determine the ethnic disparities in socioeconomic status (SES) and in receiving definitive surgical treatment and adjuvant chemotherapy and to examine if these differences contribute to ethnic disparities in survival. We studied a population-based cohort of 5131 women diagnosed with epithelial ovarian cancer at age >or=65 between 1992 and 1999, identified from the Surveillance, Epidemiology and End Results-Medicare linked databases with up to 11 years of follow-up. The percentage of women diagnosed with epithelial ovarian cancer at advanced stage (stage III or IV) was 71.6% in Caucasians and 69.7% in African-Americans. Of these 4264 with stage IC-IV disease who are recommended for chemotherapy, fewer African-Americans received chemotherapy compared to Caucasians (50.2% versus 64.7%, P < 0.001). The risk of all-cause mortality in African-Americans was not significantly different from Caucasians (hazard ratio [HR] = 1.00, 95% CI = 0.88-1.13) after controlling for patient demographics, tumor characteristics, and comorbidity. The HR remained not significant in African-Americans compared to Caucasians after additionally adjusting for treatments (0.93, 0.82-1.06) or SES (0.94, 0.82-1.08) or both (0.88, 0.77-1.01). Women who underwent cancer-directed surgery and received adjuvant chemotherapy were 50% less likely to die than those who did not. The survival benefits from these therapies were similar in Caucasian and African-American women with ovarian cancer. There was no significant difference in survival between African-American and Caucasian women with ovarian cancer after adjusting for tumor characteristics, treatment, and sociodemographic factors. Although adjuvant chemotherapy was effective in prolonging survival, substantial numbers of women with ovarian cancer still did not receive chemotherapy.


Essential oil composition, antimicrobial and cytotoxic activities of two endemic Stachys cretica subspecies (Lamiaceae) from Turkey.
The chemical compositions of the water-distilled essential oil of Stachys cretica ssp. lesbiaca Rech. fil. and S. cretica ssp. trapezuntica Rech. fil. were determined by GC and GC-MS. Altogether 63 compounds were identified. The sesquiterpene hydrocarbon, germacrene D (20.3% and 12.9% respectively) was the main component identified in both oils. Furthermore, ethanol, light petroleum, dichloromethane, ethyl acetate and n-butanol extracts prepared from the aerial parts of the plants were tested for their antimicrobial activities against six bacterial strains and the yeast Candida albicans. The extracts exhibited no antibacterial activity, but the light petroleum and n-butanolic fractions showed low antifungal activities. Crude ethanolic extracts of the two subspecies were tested for their ability to inhibit the growth of HL-60 and Ishikawa human tumor cell lines. The IC50 values were 100 microg/mL for the HL-60 cell line and 200 microg/mL for the Ishikawa cell line.


Monitoring anthrax toxin receptor dissociation from the protective antigen by NMR.
The binding of the Bacillus anthracis protective antigen (PA) to the host cell receptor is the first step toward the formation of the anthrax toxin, a tripartite set of proteins that include the enzymatic moieties edema factor (EF), and lethal factor (LF). PA is cleaved by a furin-like protease on the cell surface followed by the formation of a donut-shaped heptameric prepore. The prepore undergoes a major structural transition at acidic pH that results in the formation of a membrane spanning pore, an event which is dictated by interactions with the receptor and necessary for entry of EF and LF into the cell. We provide direct evidence using 1-dimensional (13)C-edited (1)H NMR that low pH induces dissociation of the Von-Willebrand factor A domain of the receptor capillary morphogenesis protein 2 (CMG2) from the prepore, but not the monomeric full length PA. Receptor dissociation is also observed using a carbon-13 labeled, 2-fluorohistidine labeled CMG2, consistent with studies showing that protonation of His-121 in CMG2 is not a mechanism for receptor release. Dissociation is likely caused by the structural transition upon formation of a pore from the prepore state rather than protonation of residues at the receptor PA or prepore interface.


Cancer of the oral cavity and pharynx in Karachi--identification of potential risk factors.
The objective of the study was to provide an overview of the demographics of cancer of the oral cavity and pharynx in Karachi South (1995-2001), and identify potential risk factors. Cases recorded for Karachi South, at Karachi Cancer Registry during 1(st) January 1995 to 31(st) December 2002 were analysed. For maximum completion of data cancer cases, recorded from 1(st) January 1995 to 31(st) December 2001 were included for final analysis. The age standardized incidence rates per 100000 population (ASIRs) for cancer of the oral cavity (excluding salivary gland) in Karachi South were 17.1 and 16.5 in males and females whereas the ASIRs for cancer of the pharynx (excluding nasopharynx) were 7.1 and 2.4 in males and females, respectively. The oral pharyngeal ratios were 2.4 and 6.9 for males and females and gender ratios (M F) were 1.04 for the oral cavity and 3.0 for the pharynx. The mean ages were 51 years (95% CI 49.6; 52.2) and 56.1 years (95% CI 54.4; 57.8) respectively. Cancer of the oral cavity ranked 2(nd) in Karachi in both genders. Cancer of the pharynx ranked 7(th) in males and 14(th) in females. Approximately 97% of the oral cavity and pharyngeal cancers were histologically confirmed. The majority of the oral (47.1%) and pharyngeal (51.9%) cancer cases presented as grade II lesions, and were discovered at advanced stages. Of the cancers reported during 1995-2001, 60.4% of the oral and 78.1% of the pharyngeal lesions had spread to a distant site at the time of diagnosis. Squamous cell carcinoma comprised 96.5% and 91.8% of the totals. The incidences of these cancers are comparable to the highest risk regions of the world. As distinct from other geographical areas oral cancer is as common in females as in males, which may reflect the pattern of exposure to known risk factors such as betal quid, arecanut and tobacco and the absence of alcohol as a risk factor in both genders. Apergillus contamination of arecanut could also be a risk factor but no confirmation studies or quantification is available. Despite the common risk factors, incidence of pharyngeal cancer is three times higher in men as compared with women. The keys to reducing the incidence and mortality due to oral and pharyngeal cancers are prevention and control, emphasizing cessation of tobacco use and cancer screening. However a targeted cancer and tobacco control program does not presently exist in Pakistan.


Age at diagnosis, but not HPV type, is strongly associated with clinical course in recurrent respiratory papillomatosis.
Recurrent Respiratory Papillomatosis (RRP) is a rare disease characterized by the growth of papillomas in the airway and especially the larynx. The clinical course is highly variable among individuals and there is poor understanding of the factors that drive an aggressive vs an indolent course. A convenience cohort of 339 affected subjects with papillomas positive for only HPV6 or HPV11 and clinical course data available for 1 year or more, from a large multicenter international study were included. Exploratory data analysis was conducted followed by inferential analyses with frequentist and Bayesian statistics. We examined 339 subjects: 82% were diagnosed prior to the age of 18 years, 65% were infected with HPV6, and 69% had an aggressive clinical course. When comparing age at diagnosis with clinical course, the probability of aggressiveness is high for children under five years of age then drops rapidly. For patients diagnosed after the age of 10 years, an indolent course is more common. After accounting for confounding between HPV11 and young age, HPV type was minimally associated with aggressiveness. Fast and Frugal Trees (FFTs) were utilized to determine which algorithms yield the highest accuracy to classify patients as having an indolent or aggressive clinical course and consistently created a branch for diagnostic age at ~5 years old. There was no reliable strong association between clinical course and socioeconomic or parental factors. In the largest cohort of its type, we have identified a critical age at diagnosis which demarcates a more aggressive from less aggressive clinical course.


Developing infrared spectroscopic detection for stratifying brain tumour patients: glioblastoma multiforme vs. lymphoma.
Over a third of brain tumour patients visit their general practitioner more than five times prior to diagnosis in the UK, leading to 62% of patients being diagnosed as emergency presentations. Unfortunately, symptoms are non-specific to brain tumours, and the majority of these patients complain of headaches on multiple occasions before being referred to a neurologist. As there are currently no methods in place for the early detection of brain cancer, the affected patients' average life expectancy is reduced by 20 years. These statistics indicate that the current pathway is ineffective, and there is a vast need for a rapid diagnostic test. Attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy is sensitive to the hallmarks of cancer, as it analyses the full range of macromolecular classes. The combination of serum spectroscopy and advanced data analysis has previously been shown to rapidly and objectively distinguish brain tumour severity. Recently, a novel high-throughput ATR accessory has been developed, which could be cost-effective to the National Health Service in the UK, and valuable for clinical translation. In this study, 765 blood serum samples have been collected from healthy controls and patients diagnosed with various types of brain cancer, contributing to one of the largest spectroscopic studies to date. Three robust machine learning techniques - random forest, partial least squares-discriminant analysis and support vector machine - have all provided promising results. The novel high-throughput technology has been validated by separating brain cancer and non-cancer with balanced accuracies of 90% which is comparable to the traditional fixed diamond crystal methodology. Furthermore, the differentiation of brain tumour type could be useful for neurologists, as some are difficult to distinguish through medical imaging alone. For example, the highly aggressive glioblastoma multiforme and primary cerebral lymphoma can appear similar on magnetic resonance imaging (MRI) scans, thus are often misdiagnosed. Here, we report the ability of infrared spectroscopy to distinguish between glioblastoma and lymphoma patients, at a sensitivity and specificity of 90.1% and 86.3%, respectively. A reliable serum diagnostic test could avoid the need for surgery and speed up time to definitive chemotherapy and radiotherapy.


Non-peptidergic primary afferents are presynaptic to neurokinin-1 receptor immunoreactive lamina I projection neurons in rat spinal cord.
Pain-related (nociceptive) information is carried from the periphery to the dorsal horn of the spinal cord mostly by two populations of small diameter primary afferents, the peptidergic and the non-peptidergic. The peptidergic population expresses neuropeptides, such as substance P and calcitonin gene-related peptide, while the non-peptidergic fibers are devoid of neuropeptides, express the purinergic receptor P2X3, and bind the isolectin B4 (IB4). Although it has been known for some time that in rat the peptidergic afferents terminate mostly in lamina I and outer lamina II and non-peptidergic afferents in inner lamina II, the extent of the termination of the latter population in lamina I was never investigated as it was considered as very minor. Because our preliminary evidence suggested otherwise, we decided to re-examine the termination of non-peptidergic afferents in lamina I, in particular with regards to their innervation of projection neurons expressing substance P receptors (NK-1r). We used retrograde labeling of neurons from the parabrachial nucleus combined with lectin IB4 binding and immunocytochemistry. Samples were examined by confocal and electron microscopy. By confocal microscopy, we studied the termination of non-peptidergic afferents in lamina I using IB4 binding and P2X3 immunoreactivity as markers, in relation to CGRP immunoreactivy, a marker of peptidergic afferents. The number of IB4 or P2X3-labeled fibers in lamina I was higher than previously thought, although they were less abundant than CGRP-labeled afferents. There were very few fibers double-labeled for CGRP and either P2X3 or IB4. We found a considerable number of IB4-positive fiber varicosities in close apposition to NK-1r-positive lamina I projection neurons, which were distinct from peptidergic varicosities. Furthermore, we confirmed at the ultrastructural level that there were bona fide synapses between P2X3-immunoreactive non-peptidergic boutons and neurokinin-1 receptor-positive lamina I dendrites. These results indicate the presence of direct innervation by non-peptidergic nociceptive afferents of lamina I projection neurons expressing NK-1r. Further investigations are needed to better understand the role of these connections in physiological conditions and chronic pain states.


Mechanisms of methylmercury-induced neurotoxicity: evidence from experimental studies.
Neurological disorders are common, costly, and can cause enduring disability. Although mostly unknown, a few environmental toxicants are recognized causes of neurological disorders and subclinical brain dysfunction. One of the best known neurotoxins is methylmercury (MeHg), a ubiquitous environmental toxicant that leads to long-lasting neurological and developmental deficits in animals and humans. In the aquatic environment, MeHg is accumulated in fish, which represent a major source of human exposure. Although several episodes of MeHg poisoning have contributed to the understanding of the clinical symptoms and histological changes elicited by this neurotoxicant in humans, experimental studies have been pivotal in elucidating the molecular mechanisms that mediate MeHg-induced neurotoxicity. The objective of this mini-review is to summarize data from experimental studies on molecular mechanisms of MeHg-induced neurotoxicity. While the full picture has yet to be unmasked, in vitro approaches based on cultured cells, isolated mitochondria and tissue slices, as well as in vivo studies based mainly on the use of rodents, point to impairment in intracellular calcium homeostasis, alteration of glutamate homeostasis and oxidative stress as important events in MeHg-induced neurotoxicity. The potential relationship among these events is discussed, with particular emphasis on the neurotoxic cycle triggered by MeHg-induced excitotoxicity and oxidative stress. The particular sensitivity of the developing brain to MeHg toxicity, the critical role of selenoproteins and the potential protective role of selenocompounds are also discussed. These concepts provide the biochemical bases to the understanding of MeHg neurotoxicity, contributing to the discovery of endogenous and exogenous molecules that counteract such toxicity and provide efficacious means for ablating this vicious cycle.


Respiratory syncytial virus-associated mortality in hospitalized infants and young children.
Respiratory syncytial virus (RSV) is a common cause of pediatric hospitalization, but the mortality rate and estimated annual deaths are based on decades-old data. Our objective was to describe contemporary RSV-associated mortality in hospitalized infants and children aged <2 years. We queried the Healthcare Cost and Utilization Project Kids' Inpatient Database (KID) for 2000, 2003, 2006, and 2009 and the Pediatric Health Information System (PHIS) administrative data from 2000 to 2011 for hospitalizations with International Classification of Diseases, Ninth Revision, Clinical Modification diagnosis codes for RSV infection and mortality. The KID data sets identified 607 937 RSV-associated admissions and 550 deaths (9.0 deaths/10 000 admissions). The PHIS data set identified 264 721 RSV-associated admissions and 671 deaths (25.4 deaths/10 000 admissions) (P < .001 compared with the KID data set). The 2009 KID data set estimated 42.0 annual deaths (3.0 deaths/10 000 admissions) for those with a primary diagnosis of RSV. The PHIS data set identified 259 deaths with a primary diagnosis of RSV, with mortality rates peaking at 14.0/10 000 admissions in 2002 and 2003 and decreasing to 4.0/10 000 patients by 2011 (odds ratio: 0.27 [95% confidence interval: 0.14-0.52]). The majority of deaths in both the KID and PHIS data sets occurred in infants with complex chronic conditions and in those with other acute conditions such as sepsis that could have contributed to their deaths. Deaths associated with RSV are uncommon in the 21st century. Children with complex chronic conditions account for the majority of deaths, and the relative contribution of RSV infection to their deaths is unclear.


Cerebellin-like peptide: tissue distribution in rat and guinea-pig and its release from rat cerebellum, hypothalamus and cerebellar synaptosomes in vitro.
We have developed a specific radioimmunoassay for "cerebellin", a 16-amino acid peptide recently isolated from rat cerebellum. In both rat and guinea-pig, cerebellin-like immunoreactivity was highest in the cerebellum but was also present in high concentrations elsewhere in the central nervous system, especially in the hypothalamus. In both species, cerebellin-like immunoreactivity was found in other organs (heart, kidney and stomach) and at lower concentrations in the gastrointestinal tract. In the brain of both species, cerebellin-like immunoreactivity consisted of a single molecular form with an elution position on gel filtration and high-performance liquid chromatography identical to that of synthetic rat cerebellin. However, peripheral tissue contained an additional immunoreactive peak of higher molecular weight. Cerebellin was concentrated in synaptosomal preparations of rat brain, and its subcellular distribution pattern in rat brain was identical to that of two other known synaptosomal peptides, vasoactive intestinal polypeptide and substance P. Studies with superfused cerebellar synaptosomes and slices of rat cerebellum and hypothalamus demonstrated calcium-dependent cerebellin release when stimulated by high potassium concentrations as well as the addition of the calcium ionophore A23187. Cerebellin has therefore a widespread distribution and fulfils two criteria for a neurotransmitter, in that it is found in brain synaptosomes and shows calcium dependent, depolarization-induced release from nervous tissues and isolated nerve endings. It may, therefore, be a component of a novel neurotransmitter system.


vpr deletion mutant of simian immunodeficiency virus induces AIDS in rhesus monkeys.
In previous experiments, animals infected with SIVmac239 containing a point mutation in the vpr and nef genes developed AIDS-like symptoms after early reversion of the vpr and nef genes. Here we show that two animals in which the nef gene but not the vpr gene had reverted in the first few months did not develop disease during a 3-year observation period even after reversion to a functional vpr gene 70 weeks postinfection. To study the influence of a stable vpr mutation on virus load and pathogenesis, a 43-bp deletion was introduced into the vpr gene of SIVmac239on, a nef-open mutant of SIVmac239. Four rhesus monkeys were inoculated with the vpr deletion mutant (SIV delta vpr), and two control animals were infected with SIVmac239on. Both control animals had persistent antigenemia, high cell-associated virus loads, and elevated neopterin levels. They had to be euthanized 20 and 30 weeks postinfection because of AIDS-related symptoms. However, all four rhesus monkeys inoculated with SIV delta vpr showed only transiently detectable antigenemia. The cell-associated virus loads were high in three of the four animals. Two animals with AIDS-like symptoms had to be euthanized 71 and 73 weeks postinfection. The two remaining monkeys infected with SIV delta vpr were still alive 105 weeks postinfection. In contrast to the SIVmac239on-infected animals, SIV delta vpr-infected animals had strong humoral immune responses and intermittent cellular immune responses to SIV antigens. Our data show that a functional vpr gene is not necessary for pathogenesis. However, vpr-deficient SIVmac239 variants might be slightly attenuated, allowing some animals to resist progression to disease for an extended period of time.


Correlation between plasma leptin concentration and body fat content in dogs.
To evaluate the relationship between plasma leptin concentration and body fat content in dogs. 20 spayed female Beagles that were 10 months old at the start of the experiment. Dogs were kept under regulated feeding and exercise conditions for 21 weeks, resulting in a wide range of body weights, body condition scores (BCS), and subcutaneous thicknesses. Plasma leptin concentration was measured by use of a canine leptin-specific ELISA test to evaluate its correlation to body fat content estimated by the deuterium oxide dilution method. Plasma concentrations of glucose, cholesterol, triglycerides (TG), and nonesterified fatty acids (NEFA) were also measured. Body fat content (9 to 60% of body weight) was positively and closely correlated (r = 0.920; n = 20; P < 0.001) to plasma leptin concentration (0.67 to 8.06 ng/ml), compared with other variables (ie, glucose, cholesterol, TG, and NEFA; r = 0.142, 0.412, 0.074, and 0.182, respectively). The positive relationship between plasma leptin concentration and body fat content in dogs was similar to correlations reported for humans and rodents, suggesting that plasma leptin is a quantitative marker of adiposity in dogs.


Relationship Between Diet, Tinnitus, and Hearing Difficulties.
Diet may affect susceptibility of the inner ear to noise and age-related effects that lead to tinnitus and hearing loss. This study used complementary single nutrient and dietary pattern analysis based on statistical grouping of usual dietary intake in a cross-sectional analysis of tinnitus and hearing difficulties in a large population study sample. The research was conducted using the UK Biobank resource. Tinnitus was based on report of ringing or buzzing in one or both ears that lasts more than five minutes at a time and is currently experienced at least some of the time. Identification of a hearing problem was based on self-reported difficulties with hearing. Usual dietary intake and dietary patterns (involving statistical grouping of intake to account for how foods are combined in real-life diets) were estimated based on between two and five administrations of the Oxford Web-Q 24-hour dietary recall questionnaire over the course of a year for 34,576 UK adult participants aged 40 to 69. In a multivariate model, higher intake of vitamin B12 was associated with reduced odds of tinnitus, while higher intakes of calcium, iron, and fat were associated with increased odds (B12, odds ratio [OR] 0.85, 95% confidence interval [CI] 0.75 to 0.97; Calcium, OR 1.20, 95% CI 1.08 to 1.34; Iron, OR 1.20, 95% CI 1.05 to 1.37; Fat, OR 1.33, 95% CI 1.09 to 1.62, respectively, for quintile 5 versus quintile 1). A dietary pattern characterised by high protein intake was associated with reduced odds of tinnitus (OR 0.90, 95% CI 0.82 to 0.99 for quintile 5 versus quintile 1). Higher vitamin D intake was associated with reduced odds of hearing difficulties (OR 0.90, 95% CI 0.81 to 1.00 for quintile 5 versus quintile 1), as were dietary patterns high in fruit and vegetables and meat and low in fat (Prudent diet: OR 0.89, 95% CI 0.83 to 0.96; High protein: OR 0.88, 95% CI 0.82 to 0.95; High fat: OR 1.16, 95% CI 1.08 to 1.24, respectively, for quintile 5 versus quintile 1). There were associations between both single nutrients and dietary patterns with tinnitus and hearing difficulties. Although the size of the associations was small, universal exposure for dietary factors indicates that there may be a substantial impact of diet on levels of tinnitus and hearing difficulties in the population. This study showed that dietary factors might be important for hearing health.


Precocious puberty with congenital hypothyroidism.
Precocious puberty associated with profound hypothyroidism is a rare condition. It is usually characterized by breast development, vaginal bleeding, lack of pubic hair and delayed bone age. Multicystic ovaries in profound hypothyroid patients with precocious puberty have been rarely described. Vaginal bleeding in adolescent girls should be considered as a clinical significance particularly when it is prolonged or heavy, whereas vaginal bleeding in younger girls, regardless of its duration and quantity is always of clinical importance. Bleeding in such patients could be caused by local causes such as vulvar or vaginal lesions, or it could be from the endometrium, which is usually a sign of systemic hormonal disturbance [1]. In this report a rare case of vaginal bleeding, large, multicystic ovaries, precocious puberty and delayed bone age in a 7 years old girl with profound hypothyroidism is described.


Monocular rivalry exhibits three hallmarks of binocular rivalry: evidence for common processes.
Binocular rivalry occurs when different images are presented one to each eye: the images are visible only alternately. Monocular rivalry occurs when different images are presented both to the same eye: the clarity of the images fluctuates alternately. Could both sorts of rivalry reflect the operation of a general visual mechanism for dealing with perceptual ambiguity? We report four experiments showing similarities between the two phenomena. First, we show that monocular rivalry can occur with complex images, as with binocular rivalry, and that the two phenomena are affected similarly by the size (Experiment 1) and colour (Experiment 2) of the images. Second, we show that the distribution of dominance periods during monocular rivalry has a gamma shape and is stochastic (Experiment 3). Third, we show that during periods of monocular-rivalry suppression, the threshold to detect a probe (a contrast pulse to the suppressed stimulus) is raised compared with during periods of dominance (Experiment 4). The threshold elevation is much weaker than during binocular rivalry, consistent with monocular rivalry's weak appearance. We discuss other similarities between monocular and binocular rivalry, and also some differences, concluding that part of the processing underlying both phenomena is a general visual mechanism for dealing with perceptual ambiguity.


Agricultural policy, food policy, and communicable disease policy.
Food and agricultural policy is an essential element of a communicable disease policy. The European Union has developed a more systematic and broadly based interest in questions of food safety and animal health and welfare linked to modernization of the Common Agricultural Policy, reflected in a new treaty obligation on animal welfare. Following the bovine spongiform encephalopathy crisis, moves were made to create a European competency, but implementation and enforcement resources reside with the member states. The European Animal Health Strategy is meant to lead to an EU animal health law, but this has already been constrained by fiscal austerity. The development of such a law may lead to a lowest common denominator formula that does little to enhance consumer protection or improve animal welfare. This is an inherent risk with top-down forms of Europeanization; more attention should be paid to lessons to be learned from bottom-up initiatives of the type used to counteract the bovine diarrhea virus. There will always be a tension among what is good policy for reducing the incidence of communicable disease, policy that is popular with EU citizens, and policy that is acceptable to member states.


The expression and activity of cathepsins D, H and K in asthmatic airways.
Tumstatin is an anti-angiogenic collagen IV α3 fragment, levels of which are reduced in the airways of asthmatics. Its reduction may be due to the degradation by extracellular matrix (ECM) proteases. Cathepsins play a role in ECM remodelling, with cathepsin D, H and K (CTSD, CTSH and CTSK) being associated with lung diseases. CTSD modulates the NC1 domains of collagen molecules including tumstatin, while CTSH and CTSK are involved in ECM degradation. The role of these cathepsins in the regulation of tumstatin in the lung has not previously been examined. We demonstrated that CTSB, D, F, H, K, L and S mRNA was expressed in the airways. Quantification of immunohistochemistry showed that there is no difference in the global expression of CTSD, CTSH and CTSK between asthmatics and non-asthmatics. CTSD and CTSK, but not CTSH had the capacity to degrade tumstatin. No difference was observed in the activity of CTSD and H in bronchoalveolar lavage fluid of asthmatic and non-asthmatics, while CTSK was undetectable. This indicates that while CTSD possesses the potential to directly regulate tumstatin, and thus angiogenesis through this mechanism however, it is not likely to be involved in the dysregulation of tumstatin found in asthmatic airways.


The Gln/Gln genotype of XPD codon 751 as a genetic marker for melanoma risk and Lys/Gln as an important predictor for melanoma progression: a case control study in the Swedish population.
The Xeroderma pigmentosum complementation group D (XPD) is a critical protein in the nucleotide excision repair system for DNA damage. Genetic variations in XPD exert an important effect on the capacity of DNA repair. In this study, we examined Lys751Gln polymorphism at the XPD gene in 244 melanoma patients and 251 healthy individuals (as controls) from the south-eastern region of Sweden. The associations of polymorphism with melanoma risk, as well as with melanoma features and pigment phenotypes of the melanoma patients were analysed. DNA was extracted from the mononuclear cells of venous blood of the melanoma patients and controls. XPD codon 751 was genotyped by the PCR restriction fragment length polymorphism technique. Results showed that there was no difference in the distribution of the XPD codon 751 genotypes between the melanoma patients and healthy controls. However, the Gln/Gln genotype was found to be associated with melanoma risk in the male population. Furthermore, the frequency of the Gln/Gln genotype was significantly higher in the early stages of melanomas, whereas Lys/Gln was more frequent in the later stages and in the patients with melanoma located on intermittently UV-exposed areas. No correlations between the polymorphisms and phenotypes of the patients were found. In conclusion, Gln/Gln was a useful genetic marker for melanoma risk in the males, while Lys/Gln was an important predictor for melanoma progression.


Neurofilament-L homopolymers are less mechanically stable than native neurofilaments.
Neurofilaments are cytoskeletal components of neurones that are thought to play an important structural role in the axon. Specific functions of neurofilaments are not yet well defined; however, other intermediate filaments are known to have structural and mechanical functions in different cell types. The atomic force microscope (AFM) can be used to visualize and manipulate biological structures through direct physical contact. This allows the AFM to be used to probe the mechanical properties of these structures. In this paper we present AFM images of native neurofilaments isolated from bovine spinal cord, composed of NF-L, NF-M and NF-H, and filaments polymerized in vitro from purified NF-L. Morphologically these structures, in solution and under ambient conditions, are in agreement with previous data from electron microscopy. However, repeated scanning of NF-L homopolymers (in solution) produced significant disruptions of segments of filaments, both within and at the ends of the filaments. This disruption resulted in complete loss of portions of the filaments and in breaks in the continuity of the filaments. Repeated scanning of isolated native neurofilaments under similar conditions produced no detectable structural changes. Under extremely high applied forces the native neurofilaments were bent and distorted by the action of the AFM tip, but were never broken. These data suggest that purified NF-L is not sufficient to confer complete mechanical stability to neurofilaments.


Association between inflammatory cytokines and ADHD symptoms in children and adolescents with obesity: A pilot study.
Whilst the association between Attention-Deficit/Hyperactivity Disorder (ADHD) and obesity is supported by meta-analytic evidence, the mechanisms underpinning this link need to be further elucidated. Inflammatory processes may increase the risk of ADHD symptoms in individuals with obesity. This pilot study set out to start testing this hypothesis by assessing the correlation between serum levels of inflammatory cytokines and ADHD symptoms severity in a sample of children and adolescents with obesity. We measured ADHD symptoms severity in 52 children/adolescents with obesity (BMI > 95th centile) with the Conners questionnaire, revised, short version, parent (CPRS-R:S) and teacher (CTRS-R:S) versions. Additionally, a categorical diagnosis of ADHD was established using the Kiddie-SADS-PL. Serum levels of IL-6, Il-10, and TNF-alpha were also obtained. The prevalence of ADHD was 9.6%. We found a significant correlation between IL-6, as well as TNF-alpha, and hyperactivity/impulsivity subscores of the CPRS-R:S and CTRS-R:S, that held even after controlling for BMI and oppositional symptoms. This study provides a rationale for larger, longitudinal studies to gain insight into inflammatory processes underpinning the link between obesity and ADHD. This line of research has the potential to lead to novel, pathophysiologically-based management strategies for individuals with obesity and ADHD.


Effect of chronic hypoxia on rat pulmonary resistance vessels: vasodilatation by atrial natriuretic peptide.
1. We have investigated the vasoreactivity of isolated pulmonary resistance vessels of rats after acclimatization to chronic hypoxia in a normobaric, hypoxic chamber. Vasoconstriction, in response to KCl and prostaglandin F2 alpha, and vasodilation, in response to atrial natriuretic peptide, were studied isometrically in a small-vessel myograph. Resting tensions were set to simulate transmural pressures of 17.5 mmHg or 35 mmHg. 2. There were no significant differences between intergroup internal vessel diameters or maximal contractile responses to either agonist. Both control and chronically hypoxic vessels generated significantly greater active contractions at 35 mmHg than at 17.5 mmHg. There were significant positive correlations between vessel diameter and maximum contractility for both control and chronically hypoxic vessels, but when contraction was expressed as equivalent transmural pressure no correlation existed. 3. There was a significant increase in potency (as measured by the concentration necessary to produce 50% of the maximum response) of KCl in chronically hypoxic vessels compared with control vessels at 35 mmHg, but not at 17.5 mmHg. In contrast, the potency of prostaglandin F2 alpha was significantly increased in chronically hypoxic vessels at 17.5 mmHg, but not at 35 mmHg. Thus the change in contractile responses of vessels from chronically hypoxic animals, in terms of maximal response and potency, is dependent on both resting pressure and agonist used. 4. After exposure to chronic hypoxia, atrial natriuretic peptide induced significantly greater maximal relaxation of pulmonary resistance vessels at both resting pressures, but its potency was unaffected.


The anti-obesity and health-promoting effects of tea and coffee.
This paper reviews provenance, chemical composition and properties of tea (Camelia sinensis L.) and coffee (Coffee arabica, L. and Coffeacaniphora, L.), their general health effects, as well as the currently available knowledge concerning their action on fat storage, physiological mechanisms of their effects, as well as their safety and recommended dosage for treatment of obesity. Both tea and coffee possess the ability to promote health and to prevent, to mitigate and to treat numerous disorders. This ability can be partially due to presence of caffeine in both plants. Further physiological and medicinal effects could be explained by other molecules (theaflavins, catechins, their metabolites and polyphenols in tea and polyphenol chlorogenic acid in coffee). These plants and plant molecules can be efficient for prevention and treatment of numerous metabolic disorders including metabolic syndrome, cardiovascular diseases, type 2 diabetes and obesity. Both plants and their constituents can reduce fat storage through suppression of adipocyte functions, and support of gut microbiota. In addition, tea can prevent obesity via reduction of appetite, food consumption and food absorption in gastrointestinal system and through the changes in fat metabolism.


Comparative analyses of multi-species sequences from targeted genomic regions.
The systematic comparison of genomic sequences from different organisms represents a central focus of contemporary genome analysis. Comparative analyses of vertebrate sequences can identify coding and conserved non-coding regions, including regulatory elements, and provide insight into the forces that have rendered modern-day genomes. As a complement to whole-genome sequencing efforts, we are sequencing and comparing targeted genomic regions in multiple, evolutionarily diverse vertebrates. Here we report the generation and analysis of over 12 megabases (Mb) of sequence from 12 species, all derived from the genomic region orthologous to a segment of about 1.8 Mb on human chromosome 7 containing ten genes, including the gene mutated in cystic fibrosis. These sequences show conservation reflecting both functional constraints and the neutral mutational events that shaped this genomic region. In particular, we identify substantial numbers of conserved non-coding segments beyond those previously identified experimentally, most of which are not detectable by pair-wise sequence comparisons alone. Analysis of transposable element insertions highlights the variation in genome dynamics among these species and confirms the placement of rodents as a sister group to the primates.


ACR appropriateness criteria asymptomatic patient at risk for coronary artery disease.
Atherosclerotic cardiovascular disease is the leading cause of death for both men and women in the United States. Coronary artery disease has a long asymptomatic latent period and early targeted preventive measures can reduce mortality and morbidity. It is important to accurately classify individuals at elevated risk in order to identify those who might benefit from early intervention. Imaging advances have made it possible to detect subclinical coronary atherosclerosis. Coronary artery calcium score correlates closely with overall atherosclerotic burden and provides useful prognostic information for patient management. Our purpose is to discuss use of diagnostic imaging in asymptomatic patients at elevated risk for future cardiovascular events. The goal for these patients is to further refine targeted preventative efforts based on risk. The following imaging modalities are available for evaluating asymptomatic patients at elevated risk: radiography, fluoroscopy, multidetector CT, ultrasound, MRI, cardiac perfusion scintigraphy, echocardiography, and PET. The ACR Appropriateness Criteria are evidence-based guidelines for specific clinical conditions that are reviewed every 2 years by a multidisciplinary expert panel. The guideline development and review include an extensive analysis of current medical literature from peer-reviewed journals and the application of a well-established consensus methodology (modified Delphi) to rate the appropriateness of imaging and treatment procedures by the panel. In those instances where evidence is lacking or not definitive, expert opinion may be used to recommend imaging or treatment.


Recurrence of bovine tuberculosis breakdowns in Great Britain: risk factors and prediction.
Bovine tuberculosis (bTB) is an important economic disease worldwide with implications for both animal and human health. In Great Britain the number of herds that test positive for bTB, termed "breakdowns", has increased over the last two decades. Despite more intensive testing during a breakdown, around 23% of breakdowns recur within 12 months of the previous breakdown ending, and around 38% within 24 months. These "recurrent" breakdowns may be important for onward transmission of infection. Detailed case-control data were analysed to identify factors associated with recurrence within 12 months. The model predicted 83% of all recurrent breakdowns, with a positive predictive value (PPV) of 44%. A further model, restricted to data currently available nationally, was not sufficient to predict recurrence reliably; at a sensitivity of 72-76%, the PPV was 33-36%, when validated on independent data. Factors identified to be associated with recurrence are consistent with previous studies; namely, the number of reactors, a recent bTB history in the herd and a lack of association with the confirmation status of the initial breakdown. These variables are indicative of a higher level of infection or residual infection, and could be useful in the future development of predictive models for bTB recurrence.


A Comparison of Gluteus Maximus, Biceps Femoris, and Vastus Lateralis Electromyographic Activity in the Back Squat and Barbell Hip Thrust Exercises.
The back squat and barbell hip thrust are both popular exercises used to target the lower body musculature; however, these exercises have yet to be compared. Therefore, the purpose of this study was to compare the surface electromyographic (EMG) activity of the upper and lower gluteus maximus, biceps femoris, and vastus lateralis between the back squat and barbell hip thrust. Thirteen trained women (n = 13; age = 28.9 years; height = 164 cm; mass = 58.2 kg) performed estimated 10-repetition maximums (RM) in the back squat and barbell hip thrust. The barbell hip thrust elicited significantly greater mean (69.5% vs 29.4%) and peak (172% vs 84.9%) upper gluteus maximus, mean (86.8% vs 45.4%) and peak (216% vs 130%) lower gluteus maximus, and mean (40.8% vs 14.9%) and peak (86.9% vs 37.5%) biceps femoris EMG activity than the back squat. There were no significant differences in mean (99.5% vs 110%) or peak (216% vs 244%) vastus lateralis EMG activity. The barbell hip thrust activates the gluteus maximus and biceps femoris to a greater degree than the back squat when using estimated 10RM loads. Longitudinal training studies are needed to determine if this enhanced activation correlates with increased strength, hypertrophy, and performance.


Different brain strategies underlie the categorical perception of foreign and native phonemes.
The present study using near-infrared spectroscopy examined the neuronal correlates of Japanese long/short vowel contrast discrimination and its relationship with behavioral performance by comparing native Japanese (L1) subjects and Korean subjects learning Japanese as a second language (L2). Phoneme-specific responses were predominantly observed in the left auditory area only in the L1 subjects, although the behavioral scores of the L2 subjects indicated categorical perception (CP) that was indistinguishable from that of the L1 subjects. These inconsistent relationships were more evident in the correlation coefficients between the brain recording and behavior. However, slower reaction times and non-specific brain responses in the L2 listeners suggest differences in their cortical processes from those of the L1 subjects. These findings suggest that the CP of L2 phonemes as determined by behavioral scores alone does not always predict a language-specific neural processing as employed by the L1 listeners.


Synthesis of functionalised azepanes and piperidines from bicyclic halogenated aminocyclopropane derivatives.
A series of 6,6-dihalo-2-azabicyclo[3.1.0]hexane and 7,7-dihalo-2-azabicyclo[4.1.0]heptane compounds were prepared by the reaction of dihalocarbene species with N-Boc-2,3-dihydro-1H-pyrroles or -1,2,3,4-tetrahydropyridines. Monochloro substrates were synthesised as well, using a chlorine-to-lithium exchange reaction. The behaviour of several aldehydes and ketones under reductive amination conditions with deprotected halogenated secondary cyclopropylamines was investigated, showing that this transformation typically triggers cyclopropane ring cleavage to give access to interesting nitrogen-containing ring-expanded products.


Versatile use of Azospirillum brasilense strains tagged with egfp and mCherry genes for the visualization of biofilms associated with wheat roots.
This study reports the introduction of egfp or mCherry markers to the Sp245, Sp7, and M40 wild-type strains of Azospirillum brasilense and the hhkB (encoding for a putative hybrid histidine kinase) minus mutant an isogenic strain of A. brasilense Sp245 to monitor colonization of wheat (Triticum aestivum). Two plasmids were constructed: (1) the pJMS-2 suicide plasmid derived from pSUP202 and harboring the mCherry gene expressed under the constitutive kanamycin resistance promoter to create a cis tag and (2) the broad-range plasmid pMP2449-5 that carries the mCherry gene under the lac promoter, which is derived from the plasmid pMP2444; to create the in trans tag. The stability of the plasmids encoding egfp and mCherry were confirmed in vitro for seven days of bacterial growth, and then, the A. brasilense strains harboring the plasmids were studied under nonselective conditions for adherence to seeds and, at seven or 14 days post-inoculation, for wheat root colonization. The utility of the labeled strains was proven by observation, using fluorescence microscopy and confocal laser scanning microscopy (CLSM) in wheat plants inoculated with the labeled strains and compared with the CFU g-1 for seed and wheat root. The method was suitable for observation of the in situ formation of mini-colonies, enabled visualization of bacterial colonization sites on large root fragments, and showed adherence to germinated seeds and root colonization of all strains by cell counts and direct microscopic examination. Thus, we are able to quantify the structures of the biofilms formed by each strain.


Management of healthcare waste in circumstances of limited resources: a case study in the hospitals of Nablus city, Palestine.
The objectives of this study were the assessment of healthcare waste management and the characterization of healthcare waste material generated in the hospitals in Nablus city, Palestine, and furthermore, to estimate the prevalence of hepatitis B among the cleaning personnel working in these hospitals. The medical waste generation rate in kg per bed per day was between 0.59 and 0.93 kg bed(-1) day(-1). The waste generation rate in the healthcare facilities of Nablus city hospitals was similar to some other developing countries; however, the percentage of medical wastes in the total waste stream was comparatively high. The density of medical waste at the four hospitals studied ranged between 144.9 and 188.4 kg m(-3) with a mean value of 166.7 kg m(-3). The waste segregation and handling practices were very poor. Other alternatives for waste treatment rather than incineration such as a locally made autoclave integrated with a shredder should be evaluated and implemented. The system of healthcare waste management in Nablus city is in need of immediate improvement and attention. Formulating rules and guidelines for medical waste and developing strategies for overcoming the obstacles related to waste management should be considered as an urgent matter.


Adding diverse noncanonical backbones to rosetta: enabling peptidomimetic design.
Peptidomimetics are classes of molecules that mimic structural and functional attributes of polypeptides. Peptidomimetic oligomers can frequently be synthesized using efficient solid phase synthesis procedures similar to peptide synthesis. Conformationally ordered peptidomimetic oligomers are finding broad applications for molecular recognition and for inhibiting protein-protein interactions. One critical limitation is the limited set of design tools for identifying oligomer sequences that can adopt desired conformations. Here, we present expansions to the ROSETTA platform that enable structure prediction and design of five non-peptidic oligomer scaffolds (noncanonical backbones), oligooxopiperazines, oligo-peptoids, [Formula: see text]-peptides, hydrogen bond surrogate helices and oligosaccharides. This work is complementary to prior additions to model noncanonical protein side chains in ROSETTA. The main purpose of our manuscript is to give a detailed description to current and future developers of how each of these noncanonical backbones was implemented. Furthermore, we provide a general outline for implementation of new backbone types not discussed here. To illustrate the utility of this approach, we describe the first tests of the ROSETTA molecular mechanics energy function in the context of oligooxopiperazines, using quantum mechanical calculations as comparison points, scanning through backbone and side chain torsion angles for a model peptidomimetic. Finally, as an example of a novel design application, we describe the automated design of an oligooxopiperazine that inhibits the p53-MDM2 protein-protein interaction. For the general biological and bioengineering community, several noncanonical backbones have been incorporated into web applications that allow users to freely and rapidly test the presented protocols (http://rosie.rosettacommons.org). This work helps address the peptidomimetic community's need for an automated and expandable modeling tool for noncanonical backbones.


Adaptation of chondrocytes to low oxygen tension: relationship between hypoxia and cellular metabolism.
In endochondral bone, the growth cartilage is the site of rapid growth. Since the vascular supply to the cartilage is limited, it is widely assumed that cells of the cartilage are hypoxic and that limitations in the oxygen supply regulate the energetic state of the maturing cells. In this report, we evaluate the effects of oxygen tension on chondrocyte energy metabolism, thiol status, and expression of transcription elements, HIF and AP-1. Imposition of an hypoxic environment on cultured chondrocytes caused a proportional increase in glucose utilization and elevated levels of lactate synthesis. Although we observed a statistical increase in the activities of phosphofructokinase, pyruvate kinase, lactate dehydrogenase, and creatine kinase after exposure to lowered oxygen concentrations, the effect was small. The cultured cells exhibited a decreased utilization of glutamine, possibly due to down regulation of mitochondrial function and inhibition of oxidative deamination. With respect to total energy generation, we noted that these cells are quite capable of maintaining the energy charge of the cell at low oxygen tensions. Indeed, no changes in the absolute quantity of adenine nucleotides or the energy charge ratio was observed. Hypoxia caused a decrease in the glutathione content of cultured chondrocytes and a concomitant rise in cell and medium cysteine levels. It is likely that the fall in cell glutathione level is due to decreased synthesis of the tripeptide under reduced oxygen stress and the limited supply of glutamate. The observed rise in cellular and medium cysteine levels probably reflects an increase in the rate of degradation of glutathione and a decrease in synthesis of the peptide. To explore how cells transduce these metabolic effects, gel retardation assays were used to study chondrocyte HIF and AP-1 binding activities. Chondrocyte nuclear preparations bound an HIF-oligonucleotide; however, at low oxygen tensions, no increase in HIF binding was observed. In addition, we found that AP-1 binding activities in chondrocytes exposed to low oxygen tensions was elevated, although the response was lower than that exhibited by fibroblasts exposed to the same range of oxygen concentrations. We compared these results to HIF and AP-1 binding by cells in the growth plate. There was strong HIF and AP-1 binding throughout the plate, but no evidence of selective binding to any one zone. The results of the study lend strong support to the view that chondrocytes are very well adapted to low oxygen tensions; thus, under hypoxic conditions, there is a high level of expression of both HIF and AP-1, and energy conservation appears to be near-maximum.


Family poly-victimization and cyberbullying among adolescents in a Chinese school sample.
The sustained increase in their use of social networking facilitates the development of adolescents but comes with the risk of cyberbullying, which creates new challenges in regard to adolescent protection. Past evidence shows that family victimization may play an essential role in the way adolescents learn cyberbullying behaviors. Yet, research on the co-occurrence of family victimization and cyberbullying is limited. This study aims to investigate the associations between cyberbullying and family victimization among adolescents, and to examine the health correlates of cyberbullying and family poly-victimization. A large sample of 18,341 students, aged 15-17, from six cities in China, collected between 2009 and 2010 is employed in the present study, which investigated the association between various kinds of family victimization and adolescent cyberbullying. Data analysis was conducted in 2017. In-law conflict, intimate partner violence, elder abuse and neglect, and child maltreatment were associated with a higher possibility of children becoming internet victims. Parents' divorce and separation, low family income, mother's low level of education, and father's unemployment were all associated with cyberbullying victimization. Cyber victimization was positively correlated to symptoms of PTSD and depression, self-harm, and other physical and mental health variables. Possible explanations for the relationships found in this study are discussed and implications for future research and services are provided. Proactive screening for family poly-victimization and cyberbullying is suggested. Schools are highly recommended to cooperate with parents to promote cyber safety.


Validated and longitudinally stable asthma phenotypes based on cluster analysis of the ADEPT study.
Asthma is a disease of varying severity and differing disease mechanisms. To date, studies aimed at stratifying asthma into clinically useful phenotypes have produced a number of phenotypes that have yet to be assessed for stability and to be validated in independent cohorts. The aim of this study was to define and validate, for the first time ever, clinically driven asthma phenotypes using two independent, severe asthma cohorts: ADEPT and U-BIOPRED. Fuzzy partition-around-medoid clustering was performed on pre-specified data from the ADEPT participants (n = 156) and independently on data from a subset of U-BIOPRED asthma participants (n = 82) for whom the same variables were available. Models for cluster classification probabilities were derived and applied to the 12-month longitudinal ADEPT data and to a larger subset of the U-BIOPRED asthma dataset (n = 397). High and low type-2 inflammation phenotypes were defined as high or low Th2 activity, indicated by endobronchial biopsies gene expression changes downstream of IL-4 or IL-13. Four phenotypes were identified in the ADEPT (training) cohort, with distinct clinical and biomarker profiles. Phenotype 1 was "mild, good lung function, early onset", with a low-inflammatory, predominantly Type-2, phenotype. Phenotype 2 had a "moderate, hyper-responsive, eosinophilic" phenotype, with moderate asthma control, mild airflow obstruction and predominant Type-2 inflammation. Phenotype 3 had a "mixed severity, predominantly fixed obstructive, non-eosinophilic and neutrophilic" phenotype, with moderate asthma control and low Type-2 inflammation. Phenotype 4 had a "severe uncontrolled, severe reversible obstruction, mixed granulocytic" phenotype, with moderate Type-2 inflammation. These phenotypes had good longitudinal stability in the ADEPT cohort. They were reproduced and demonstrated high classification probability in two subsets of the U-BIOPRED asthma cohort. Focusing on the biology of the four clinical independently-validated easy-to-assess ADEPT asthma phenotypes will help understanding the unmet need and will aid in developing tailored therapies. NCT01274507 (ADEPT), registered October 28, 2010 and NCT01982162 (U-BIOPRED), registered October 30, 2013.


Dose-dependent in vitro effect of recombinant human DNase on rheological and transport properties of cystic fibrosis respiratory mucus.
Recombinant human deoxyribonuclease (rhDNase) has been demonstrated to reduce the viscosity of purulent cystic fibrosis (CF) respiratory mucus, to improve pulmonary function and to reduce the risk of respiratory tract infectious exacerbations, but its effect on mucus transportability has not so far been investigated. The dose-dependent effect of rhDNase was analysed in vitro on mucus transport rate (tr) by ciliary activity and by simulated cough (cough transport (ct)), as well as on mucus viscosity and surface properties. Purulent CF sputa (n = 15) were incubated for 30 min at 37 degrees C with either rhDNase at three different concentrations (final concentrations 0.2, 2 or 20 micrograms.ml-1 of mucus) or placebo. No significant dose-dependent effect of rhDNase on the mucociliary transport rate was observed when the samples wer statistically analysed together. However, in the larger group of mucus samples (n = 11) with a low initial mucociliary transport rate, the latter was improved at each rhDNase concentration (tr0.2 = 0.69, tr2 = 0.88 and tr20 = 0.87) as compared to placebo (trp = 0.58). In the smaller group of mucus samples (n = 4) with high initial transport rate, a decrease in mucociliary transport rate was observed, particularly at the highest concentration rhDNase assayed, i.e. 20 micrograms.ml-1 of mucus (tr20 = 0.58) as compared to placebo (trp = 0.86). The mucus cough transport was increased by rhDNase (ct0.2 = 25 mm, ct2 = 27.5 mm and ct20 = 31 mm) as compared to placebo (ctp = 23.5 mm).(ABSTRACT TRUNCATED AT 250 WORDS)


Cellular pathways controlling integron cassette site folding.
By mobilizing small DNA units, integrons have a major function in the dissemination of antibiotic resistance among bacteria. The acquisition of gene cassettes occurs by recombination between the attI and attC sites catalysed by the IntI1 integron integrase. These recombination reactions use an unconventional mechanism involving a folded single-stranded attC site. We show that cellular bacterial processes delivering ssDNA, such as conjugation and replication, favour proper folding of the attC site. By developing a very sensitive in vivo assay, we also provide evidence that attC sites can recombine as cruciform structures by extrusion from double-stranded DNA. Moreover, we show an influence of DNA superhelicity on attC site extrusion in vitro and in vivo. We show that the proper folding of the attC site depends on both the propensity to form non-recombinogenic structures and the length of their variable terminal structures. These results draw the network of cell processes that regulate integron recombination.


A recently developed bifacial platelet-rich fibrin matrix.
Platelet-rich plasma (PRP) is used clinically in liquid or gel form to promote tissue repair. Because of the poor mechanical properties, conventional PRP is often difficult to handle when used in clinical settings and requires secure implantation in a specific site, otherwise when released growth factors could be washed out during an operation. In this study, we analyzed the end product of a recently developed commercially available system (FIBRINET), which is a dense pliable, platelet-rich fibrin matrix (PRFM). We characterized the mechanical properties of PRFM and tested whether PRFM releases growth factors and whether released factors induce the proliferation of mesenchymal stem cells (MSC). Mechanical properties as well as platelet distribution were evaluated in PRFM. PRFM demonstrated robust mechanical properties, with a tear elastic modulus of 937.3 +/- 314.6 kPa, stress at a break of 1476.0 +/- 526.3 kPa, and an elongation at break of 146.3 +/- 33.8 %. PRFM maintained its mechanical properties throughout the testing process. Microscopic observations showed that the platelets were localized on one side of the matrix. Elevated levels of PDGF-AA, PDGF-AB, EGF, VEGF, bFGF and TGF-beta1 were measured in the day 1-conditioned media (CM) of PRFM and growth factor levels decreased thereafter. BMP2 and BMP7 were not detectable. MSC culture media supplemented with 20% PRFM-CM stimulated MSC cell proliferation; at 24 and 48 hours the induction of the proliferation was significantly greater than the induction obtained with media supplemented with 20% foetal bovine serum. The present study shows that the production of a dense, physically robust PRFM made through high-speed centrifugation of intact platelets and fibrin in the absence of exogenous thrombin yields a potential tool for accelerating tissue repair.


Recombinant vaccinia viruses for the characterization of Plasmodium falciparum-specific cytotoxic T lymphocytes: recognition of processed antigen despite limited re-stimulation efficacy.
Cytotoxic T lymphocytes (CTL) have been implicated in immunity to Plasmodium falciparum infection and disease. We have previously described the use of peptides to define malaria-specific CTL epitopes. To determine whether these peptide epitopes are processed intracellularly from the whole antigen we have developed recombinant vaccinia viruses (rVV) expressing three malaria antigens: thrombospondin-related adhesive protein (TRAP), Pfs16 and the C-terminal half of liver-stage antigen (LSA)-1. Target cells infected with recombinant viruses were lysed by malaria-specific CTL from semi-immune African donors. We also tested the ability of cells infected with these recombinant vaccinia viruses to re-stimulate malaria-specific CTL in peripheral blood lymphocytes from malaria immune adults. Two other pox virus recombinants, NYVAC, an attenuated vaccinia virus, and ALVAC, a canarypox virus, both expressing malaria antigens were also evaluated for their ability to stimulate malaria-specific CTL in contrast to peptide, none of these viruses successfully re-stimulated CTL from the peripheral blood lymphocytes of semi-immune donors. The ability of human CTL from naturally exposed individuals to recognize processed antigen supports the relevance of these cells in protective immunity to malaria.


Structure and kinetic characterization of human sperm-specific glyceraldehyde-3-phosphate dehydrogenase, GAPDS.
hGAPDS (human sperm-specific glyceraldehyde-3-phosphate dehydrogenase) is a glycolytic enzyme essential for the survival of spermatozoa, and constitutes a potential target for non-hormonal contraception. However, enzyme characterization of GAPDS has been hampered by the difficulty in producing soluble recombinant protein. In the present study, we have overexpressed in Escherichia coli a highly soluble form of hGAPDS truncated at the N-terminus (hGAPDSΔN), and crystallized the homotetrameric enzyme in two ligand complexes. The hGAPDSΔN-NAD+-phosphate structure maps the two anion-recognition sites within the catalytic pocket that correspond to the conserved Ps site and the newly recognized Pi site identified in other organisms. The hGAPDSΔN-NAD+-glycerol structure shows serendipitous binding of glycerol at the Ps and new Pi sites, demonstrating the propensity of these anion-recognition sites to bind non-physiologically relevant ligands. A comparison of kinetic profiles between hGAPDSΔN and its somatic equivalent reveals a 3-fold increase in catalytic efficiency for hGAPDSΔN. This may be attributable to subtle amino acid substitutions peripheral to the active centre that influence the charge properties and protonation states of catalytic residues. Our data therefore elucidate structural and kinetic features of hGAPDS that might provide insightful information towards inhibitor development.


Comparative analysis of showering protocols for mass-casualty decontamination.
A well-established provision for mass-casualty decontamination that incorporates the use of mobile showering units has been developed in the UK. The effectiveness of such decontamination procedures will be critical in minimizing or preventing the contamination of emergency responders and hospital infrastructure. The purpose of this study was to evaluate three empirical strategies designed to optimize existing decontamination procedures: (1) instructions in the form of a pictorial aid prior to decontamination; (2) provision of a washcloth within the showering facility; and (3) an extended showering period. The study was a three-factor, between-participants (or "independent") design with 90 volunteers. The three factors each had two levels: use of washcloths (washcloth/no washcloth), washing instructions (instructions/no instructions), and shower cycle duration (three minutes/six minutes). The effectiveness of these strategies was quantified by whole-body fluorescence imaging following application of a red fluorophore to multiple, discrete areas of the skin. All five showering procedures were relatively effective in removing the fluorophore "contaminant", but the use of a cloth (in the absence of instructions) led to a significant ( appox. 20%) improvement in the effectiveness of decontamination over the standard protocol (p <0.05). Current mass-casualty decontamination effectiveness, especially in children, can be optimized by the provision of a washcloth. This simple but effective approach indicates the value of performing controlled volunteer trials for optimizing existing decontamination procedures.


Suicide risk and firearm ownership and storage behavior in a large military sample.
Firearms account for the majority of suicide deaths in the United States military and access to firearms is a risk factor for suicide. Prior research has shown service members tend to store firearms unsafely, with some research indicating this is particularly true among those with elevated suicide risk. Existing research has focused on individuals at known risk for suicide; however, those who die by suicide using a firearm are prone to avoiding mental healthcare and underreporting suicidal ideation, thereby necessitating an understanding of this phenomenon among firearm owners outside of the mental healthcare system. The present study examined firearm storage and suicide risk in a large nonclinical sample of service members (total sample n = 953; firearm owning sample = 473). Lifetime suicidal ideation, current depressive symptoms, and perceived likelihood of making a future suicide attempt were associated with unsafe firearm storage. In contrast, lifetime suicidal ideation was not associated with a greater likelihood to own firearms. These findings suggest those at risk of suicide are more likely to store firearms unsafely, which increases ease of access to firearms. These findings reiterate the importance of means safety as a suicide prevention strategy.


Glucocorticoid inhibition of gene expression and proliferation of murine lymphoid cells in vitro.
Glucocorticoids inhibit the proliferation of murine T-lymphoma P1798 cells. P1798 cells do not die in the presence of dexamethasone, and the process of inhibition of proliferation is completely reversible. As cells cease to divide, expression of a number of genes is inhibited. Among these are genes the expression of which is regulated in some manner that is linked to cell proliferation. We have undertaken to study the mechanism whereby glucocorticoids inhibit the expression of genes in P1798 cells. Three model systems will be reviewed. In all cases, these appear to be examples of secondary regulation. Glucocorticoid-mediated inhibition of transcription of the DNA encoding ribosomal RNA (rDNA) has been investigated in some detail. The data indicate that dexamethasone causes a decrease in the amount or activity of an RNA polymerase I transcription initiation factor. This factor exhibits a short biological half-life and the data are consistent with the hypothesis that glucocorticoids regulate the synthesis of this transcription factor. The gene encoding thymidine kinase appears to be regulated in a similar fashion. On this basis, we propose that glucocorticoids may have the general property of regulating the synthesis of certain transcription factors. Glucocorticoids also regulate the translation of a certain class of mRNAs, including those that encode ribosomal proteins. These are characterized by a low efficiency of translation in untreated cells. Upon exposure to dexamethasone, the translation of these mRNAs is disproportionately inhibited. We speculate that translation of the mRNAs encoding certain transcription factors may be regulated in a similar fashion. Specifically, we propose that transcription of certain proliferation-related genes may be dependent upon factors of short biological half-life. These are encoded by mRNAs that are poorly translated under optimal growth conditions. Any slight perturbation in translation efficiency, as caused by glucocorticoids, results in a disproportionate inhibition of synthesis of these hypothetical transcription factors. Transcription of a class of proliferation-related genes ceases as a result.


Obesity/Type II diabetes alters macrophage polarization resulting in a fibrotic tendon healing response.
Type II Diabetes (T2DM) dramatically impairs the tendon healing response, resulting in decreased collagen organization and mechanics relative to non-diabetic tendons. Despite this burden, there remains a paucity of information regarding the mechanisms that govern impaired healing of diabetic tendons. Mice were placed on either a high fat diet (T2DM) or low fat diet (lean) and underwent flexor tendon transection and repair surgery. Healing was assessed via mechanical testing, histology and changes in gene expression associated with collagen synthesis, matrix remodeling, and macrophage polarization. Obese/diabetic tendons healed with increased scar formation and impaired mechanical properties. Consistent with this, prolonged and excess expression of extracellular matrix (ECM) components were observed in obese/T2DM tendons. Macrophages are involved in both inflammatory and matrix deposition processes during healing. Obese/T2DM tendons healed with increased expression of markers of pro-inflammatory M1 macrophages, and elevated and prolonged expression of M2 macrophages markers that are involved in ECM deposition. Here we demonstrate that tendons from obese/diabetic mice heal with increased scar formation and increased M2 polarization, identifying excess M2 macrophage activity and matrix synthesis as a potential mechanism of the fibrotic healing phenotype observed in T2DM tendons, and as such a potential target to improve tendon healing in T2DM.


Learning interpretable SVMs for biological sequence classification.
Support Vector Machines (SVMs)--using a variety of string kernels--have been successfully applied to biological sequence classification problems. While SVMs achieve high classification accuracy they lack interpretability. In many applications, it does not suffice that an algorithm just detects a biological signal in the sequence, but it should also provide means to interpret its solution in order to gain biological insight. We propose novel and efficient algorithms for solving the so-called Support Vector Multiple Kernel Learning problem. The developed techniques can be used to understand the obtained support vector decision function in order to extract biologically relevant knowledge about the sequence analysis problem at hand. We apply the proposed methods to the task of acceptor splice site prediction and to the problem of recognizing alternatively spliced exons. Our algorithms compute sparse weightings of substring locations, highlighting which parts of the sequence are important for discrimination. The proposed method is able to deal with thousands of examples while combining hundreds of kernels within reasonable time, and reliably identifies a few statistically significant positions.


L-Carnitine potentiates the anti-inflammatory and antinociceptive effects of diclofenac sodium in an experimentally-induced knee osteoarthritis rat model.
The aim of the present research is to investigate the efficacy of L-carnitine (LC) as a complementary therapy to diclofenac sodium (Dic) treatment in a mono-iodoacetate (MIA) induced knee osteoarthritis (OA) rat model, with respect to pain relief and the underlying pathology. Fifty adult male albino rats were randomly divided into five groups (n=10): Control, OA, OA/Dic, OA/LC, and OA/Dic+LC. Knee diameter and pain assessment tests were done weekly. After four weeks, serum malondialdehyde, reduced glutathione, interleukin 1-β, tumor necrosis factor-alpha, prostaglandin E2, and bone-specific alkaline phosphatase were measured. The injected knees were removed and processed for the histological and immunohistological study of matrix metalloproteinase-13 (MMP-13) and cyclooxygenase 2 (COX-2). Also, histological examination of dorsal root ganglia and calcitonin gene-related peptide (CGRP) expression in the spinal cord were assessed. Treatment with Dic and/or LC significantly reduced knee swelling, improved pain-related behaviors, inflammatory and oxidative stress markers, attenuated the MIA-mediated histopathological alteration in the knee joint, and down-regulated expression of MMP-13 and COX-2 in the knee joint. It, also, significantly reduced CGRP expression, compared with the OA group. Dic+LC showed a better effect in improving some parameters than each treatment alone. LC plus Dic is a more effective therapy than Dic alone for OA treatment.


Interferon-gamma- and interleukin-4-targeted gene therapy for atopic allergic disease.
Two cytokines, interferon-gamma (IFN-gamma) and interleukin-4 (IL-4), which play critical roles in the regulation of serum IgE level by directing the interplay of T helper (Th)1 and Th2 cells, were chosen as targets for gene therapy. Anti-allergic activity was evaluated by determining the serum IgE level, and the functional status of each helper T cell was monitored by the serum concentrations of IgG1 and IgG2a. Experimental animals (BALB/c mice) were divided into four groups: the control group; the ovalbumin (OVA) group; the IFN-gamma group; and the IL-4 group. The control group was injected with saline and the OVA group with OVA-alum. The IFN-gamma and IL-4 groups were treated with OVA-alum plus the cDNAs of mouse IFN-gamma and IL-4 in an expression vector. These treatments were applied intramuscularly on a monthly basis for 4 months. OVA-alum treatment significantly increased the serum IgE and IgG1 concentrations, but did not affect IgG2a. Concomitant treatments with the cDNA of IFN-gamma or IL-4 returned the serum IgE almost to the control level and significantly suppressed the OVA-induced increase of IgG1. IFN-gamma cDNA increased the serum IgG2a but IL-4 cDNA had no affect. These results suggest that IFN-gamma inhibited the OVA-induced IgE production by suppressing the Th2 pathway and by enhancing the Th1 pathway. Administration of IL-4 cDNA suppressed the OVA-induced enhancement of IgE production by inhibiting the Th2 pathway rather than by potentiating it.


Variation in gene expression within clones of the earthworm Dendrobaena octaedra.
Gene expression is highly plastic, which can help organisms to both acclimate and adapt to changing environments. Possible variation in gene expression among individuals with the same genotype (among clones) is not widely considered, even though it could impact the results of studies that focus on gene expression phenotypes, for example studies using clonal lines. We examined the extent of within and between clone variation in gene expression in the earthworm Dendrobaena octaedra, which reproduces through apomictic parthenogenesis. Five microsatellite markers were developed and used to confirm that offspring are genetic clones of their parent. After that, expression of 12 genes was measured from five individuals each from six clonal lines after exposure to copper contaminated soil. Variation in gene expression was higher over all genotypes than within genotypes, as initially assumed. A subset of the genes was also examined in the offspring of exposed individuals in two of the clonal lines. In this case, variation in gene expression within genotypes was as high as that observed over all genotypes. One gene in particular (chymotrypsin inhibitor) also showed significant differences in the expression levels among genetically identical individuals. Gene expression can vary considerably, and the extent of variation may depend on the genotypes and genes studied. Ensuring a large sample, with many different genotypes, is critical in studies comparing gene expression phenotypes. Researchers should be especially cautious inferring gene expression phenotypes when using only a single clonal or inbred line, since the results might be specific to only certain genotypes.


Recreational use in dispersed public lands measured using social media data and on-site counts.
Outdoor recreation is one of many important benefits provided by public lands. Data on recreational use are critical for informing management of recreation resources, however, managers often lack actionable information on visitor use for large protected areas that lack controlled access points. The purpose of this study is to explore the potential for social media data (e.g., geotagged images shared on Flickr and trip reports shared on a hiking forum) to provide land managers with useful measures of recreational use to dispersed areas, and to provide lessons learned from comparing several more traditional counting methods. First, we measure daily and monthly visitation rates to individual trails within the Mount Baker-Snoqualmie National Forest (MBSNF) in western Washington. At 15 trailheads, we compare counts of hikers from infrared sensors, timelapse cameras, and manual on-site counts, to counts based on the number of shared geotagged images and trip reports from those locations. Second, we measure visitation rates to each National Forest System (NFS) unit across the US and compare annual measurements derived from the number of geotagged images to estimates from the US Forest Service National Visitor Use Monitoring Program. At both the NFS unit and the individual-trail scales, we found strong correlations between traditional measures of recreational use and measures based on user-generated content shared on the internet. For national forests in every region of the country, correlations between official Forest Service statistics and geotagged images ranged between 55% and 95%. For individual trails within the MBSNF, monthly visitor counts from on-site measurements were strongly correlated with counts from geotagged images (79%) and trip reports (91%). The convenient, cost-efficient and timely nature of collecting and analyzing user-generated data could allow land managers to monitor use over different seasons of the year and at sites and scales never previously monitored, contributing to a more comprehensive understanding of recreational use patterns and values.


Treatment of small hepatocellular carcinomas.
There is growing interest in screening to detect symptomless hepatocellular carcinoma (HCC), which should be easier to treat than symptomatic tumours. Combined alpha-fetoprotein and ultrasound monitoring can detect HCCs of 1 cm, and Lipiodol retention can be detected in tumours smaller than 1 cm. A number of treatment options are available. Surgical resection may be curative in selected patients with a single small tumour, but the cirrhotic patient is left with a diseased liver and the risk of tumour recurrence or death from underlying liver dysfunction. Orthotopic liver transplantation is a rational treatment for patients with decompensating cirrhosis and a small HCC, but it is expensive and necessitates immunosuppression. A variety of targeted or local therapies, either individually or in combination, can be used to treat HCC. These include percutaneous alcohol injection into an HCC, which may be an alternative to surgical resection. Tumour necrosis can be seen after targeted Lipiodol chemotherapy or radiotherapy. Transcatheter arterial embolisation selectively embolises the feeding artery, and can be combined with Lipiodol chemotherapy. Small tumours are thus amenable to treatment, even in patients who cannot have surgery. Screening and treatment for symptomless HCC seems justified, unless controlled trials teach us differently.