Comprehensive whole-genome sequencing of an early-stage primary myelofibrosis patient defines low mutational burden and non-recurrent candidate genes.
In order to identify novel somatic mutations associated with classic BCR/ABL1-negative myeloproliferative neoplasms, we performed high-coverage genome sequencing of DNA from peripheral blood granulocytes and cultured skin fibroblasts from a patient with MPL W515K-positive primary myelofibrosis. The primary myelofibrosis genome had a low somatic mutation rate, consistent with that observed in similar hematopoietic tumor genomes. Interfacing of whole-genome DNA sequence data with RNA expression data identified three somatic mutations of potential functional significance: i) a nonsense mutation in CARD6, implicated in modulation of NF-kappaB activation; ii) a 19-base pair deletion involving a potential regulatory region in the 5'-untranslated region of BRD2, implicated in transcriptional regulation and cell cycle control; and iii) a non-synonymous point mutation in KIAA0355, an uncharacterized protein. Additional mutations in three genes (CAP2, SOX30, and MFRP) were also evident, albeit with no support for expression at the RNA level. Re-sequencing of these six genes in 178 patients with polycythemia vera, essential thrombocythemia, and myelofibrosis did not identify recurrent somatic mutations in these genes. Finally, we describe methods for reducing false-positive variant calls in the analysis of hematologic malignancies with a low somatic mutation rate. This trial is registered with ClinicalTrials.gov (NCT01108159).


Oral bioavailability of cinnarizine in dogs: relation to SNEDDS droplet size, drug solubility and in vitro precipitation.
The in vivo performance of self-nanoemulsifying drug delivery systems (SNEDDSs) with different in vitro physicochemical properties were determined with the purpose of elucidating the parameters determining the in vivo performance of SNEDDSs. The in vitro characterisation included the use of pulsed field gradient NMR and the dynamic lipolysis model. In vivo characterisation was carried out in dogs with elevated gastric pH. Four SNEDDSs containing cinnarizine were dosed orally, and the obtained PK profiles were related to in vitro characterisation data. The SNEDDSs with the lowest solubility of cinnarizine in the preconcentrates and the smallest droplet size had the highest AUC values after oral administration. No difference in C(max) and t(max) was observed between the SNEDDSs. Despite of precipitation occurring during in vitro lipolysis of one of the SNEDDS this SNEDDS performed as well in vivo as another SNEDDS that did not show any precipitation. The area under the colloidal dispersion curves as well as under the lipolysis curves could be used to rank order the in vivo performance of the SNEDDSs. Selection of in vitro optimisation parameters for SNEDDSs should be done carefully. It may not always be best to aim for the highest solubility in the preconcentrate and to avoid precipitation during in vitro lipolysis.


Atypical antipsychotic poisoning in young children: a multicentre analysis of poisons centres data.
Although paediatric patients frequently suffer from intoxications with atypical antipsychotics, the number of studies in young children, which have assessed the effects of acute exposure to this class of drugs, is very limited. The aim of this study was to achieve a better characterization of the acute toxicity profile in young children of the atypical antipsychotics clozapine, olanzapine, quetiapine, and risperidone. We performed a multicentre retrospective analysis of cases with atypical antipsychotics intoxication in children younger than 6 years, reported by physicians to German, Austrian, and Swiss Poisons Centres for the 9-year period between January 1, 2001 and December 31, 2009. One hundred and six cases (31 clozapine, 29 olanzapine, 12 quetiapine, and 34 risperidone) were available for analysis. Forty-seven of the children showed minor, 28 moderate, and 2 severe symptoms. Twenty-nine cases were asymptomatic. No fatalities were recorded. Symptoms predominantly involved the central nervous and cardiovascular systems. Minor reduction in vigilance (Glasgow Coma Scale score >9) (62 %) was the most frequently reported symptom, followed by miosis (12 %) and mild tachycardia (10 %). Extrapyramidal motor symptoms were observed in one case (1 %) after ingestion of risperidone. In most cases, surveillance and supportive care were sufficient to achieve a good outcome, and all children made full recovery. Paediatric antipsychotic exposure can result in significant poisoning; however, in most cases only minor or moderate symptoms occurred and were followed by complete recovery. Symptomatic patients should be monitored for central nervous system depression and an electrocardiogram should be obtained.


Animal Functional Magnetic Resonance Imaging: Trends and Path Toward Standardization.
Animal whole-brain functional magnetic resonance imaging (fMRI) provides a non-invasive window into brain activity. A collection of associated methods aims to replicate observations made in humans and to identify the mechanisms underlying the distributed neuronal activity in the healthy and disordered brain. Animal fMRI studies have developed rapidly over the past years, fueled by the development of resting-state fMRI connectivity and genetically encoded neuromodulatory tools. Yet, comparisons between sites remain hampered by lack of standardization. Recently, we highlighted that mouse resting-state functional connectivity converges across centers, although large discrepancies in sensitivity and specificity remained. Here, we explore past and present trends within the animal fMRI community and highlight critical aspects in study design, data acquisition, and post-processing operations, that may affect the results and influence the comparability between studies. We also suggest practices aimed to promote the adoption of standards within the community and improve between-lab reproducibility. The implementation of standardized animal neuroimaging protocols will facilitate animal population imaging efforts as well as meta-analysis and replication studies, the gold standards in evidence-based science.


Ectopic corticotropin-releasing hormone (CRH) syndrome from metastatic small cell carcinoma: a case report and review of the literature.
Cushing's Syndrome (CS) which is caused by isolated corticotropin-releasing hormone (CRH) production, rather than adrenocorticotropin (ACTH) production, is extremely rare. We describe the clinical presentation, course, laboratory values and pathologic findings of a patient with isolated ectopic CRH causing CS. We review the literature of the types of tumors associated with this unusual syndrome and the behavior of these tumors by endocrine testing. A 56 year old woman presented with clinical and laboratory features consistent with ACTH-dependent CS. Pituitary imaging was normal and cortisol did not suppress with a high dose dexamethasone test, consistent with a diagnosis of ectopic ACTH. CT imaging did not reveal any discrete lung lesions but there were mediastinal and abdominal lymphadenopathy and multiple liver lesions suspicious for metastatic disease. Laboratory testing was positive for elevated serum carcinoembryonic antigen and the neuroendocrine marker chromogranin A. Serum markers of carcinoid, medullary thyroid carcinoma, and pheochromocytoma were in the normal range. Because the primary tumor could not be identified by imaging, biopsy of the presumed metastatic liver lesions was performed. Immunohistochemistry was consistent with a neuroendocrine tumor, specifically small cell carcinoma. Immunostaining for ACTH was negative but was strongly positive for CRH and laboratory testing revealed a plasma CRH of 10 pg/ml (normal 0 to 10 pg/ml) which should have been suppressed in the presence of high cortisol. This case illustrates the importance of considering the ectopic production of CRH in the differential diagnosis for presentations of ACTH-dependent Cushing's Syndrome.


Temporal Trends in the Incidence and Natural History of Diverticulitis: A Population-Based Study.
Data on the incidence and natural history of diverticulitis are largely hospital-based and exclude the majority of diverticulitis patients, who are treated in an outpatient setting for uncomplicated diverticulitis. We assessed temporal trends in the epidemiology of diverticulitis in the general population. Through the Rochester Epidemiology Project we reviewed the records of all individuals with a diagnosis of diverticulitis from 1980 to 2007 in Olmsted County, Minnesota, USA. In 1980-1989, the incidence of diverticulitis was 115/100,000 person-years, which increased to 188/100,000 in 2000-2007 (P<0.001). Incidence increased with age (P<0.001); however, the temporal increase was greater in younger people (P<0.001). Ten years after the index and second diverticulitis episodes, 22% and 55% had a recurrence, respectively. This recurrence rate was greater in younger people (hazard ratio (HR) per decade 0.63; 95% confidence interval (CI), 0.59-0.66) and women (HR 0.68; 95% CI, 0.58-0.80). Complications were seen in 12%; this rate did not change over time. Recurrent diverticulitis was associated with a decreased risk of complications (P<0.001). Age was associated with increased risk of local (odds ratio (OR) 1.27 per decade; 95% CI, 1.04-1.57) and systemic (OR 1.83; 95% CI, 1.20-2.80) complications. Survival after diverticulitis was lower in older people (P<0.001) and men (P<0.001) and worsened over time (P<0.001). The incidence of surgery for diverticulitis did not change from 1980 to 2007. The incidence of diverticulitis has increased by 50% in 2000-2007 compared with 1990-1999, and more so in younger people. Complications are relatively uncommon. Recurrent diverticulitis is frequent but typically uncomplicated. Younger people with diverticulitis have less severe disease, more recurrence, and better survival.


The impact of parental migration on the mental health of children left behind.
This study aims to examine the impact of parental international migration on the mental health of children left behind. Data for this paper were taken from the "Child Health and Migrant Parents in South-East Asia, Thailand, 2008" project. A total of 1,030 children (519 from emigrant parents and 511 from non-migrant parents) were covered. Data were collected from the caregivers of children using the Strength and Difficulties Questionnaire. Multivariate analyses were performed to determine the association between parental migration and the mental health status of children left behind. Several factors were significantly associated with children's mental health. Our study did not find any association between current parental migration status and mental health status of the children left behind. The study found, however, that mother's earlier migration history had a significant, independent association with mental health problems of the children left behind. Therefore, effective strategies to prevent such mental health problems among children are warranted.


Sequence context effects in DNA replication blocks induced by aflatoxin B1.
The genotoxic effects of the potent mutagenic carcinogen aflatoxin B1 (AFB1) are believed to be mediated by its reaction with the N-7 atom of guanine residues in DNA. We have analyzed the effect of AFB1-induced chemical modification on the template function of single-stranded DNA in vitro. The experimental strategy involves the elongation of a primer on a modified template by Escherichia coli DNA polymerase I (large fragment) and analysis of the products by high-resolution gel electrophoresis. Our data show that (i) AFB1 induces specific replication blocks one nucleotide 3' to the sites of occurrence of guanine residues on template DNA; (ii) AFB1-induced replication blocks occur predominantly at sequences capable of participation in intrastrand base pairing; (iii) within the intrastrand base-paired regions there are strong sequence context effects, in accordance with the previously described [Muench, K. F., Misra, R. P. & Humayun, M. Z. (1983) Proc. Natl. Acad. Sci. USA 80, 6-10] specificity "rules" that apply to the reaction of AFB1 with guanine residues in double-stranded DNA; (iv) there is evidence that the (7-guanyl)-AFB1 adducts as well as secondary derivatives such as the formamidopyrimidine-AFB1 act as replication blocks. In summary, these data suggest that previously observed inhibition of DNA replication and transcription by AFB1 is directly attributable to (7-guanyl)-AFB1 adducts or their secondary reaction products.


Stress effects on rats chronically receiving a highly palatable diet are sex-specific.
The stress response is known to lead to behavioral and metabolic changes. Exposure to chronic stress can promote the development of physiological and behavioral dysfunctions, including alterations in feeding behavior. The aim of this study was to verify whether chronic restraint stress alters the consumption of a highly palatable, highly caloric diet (chocolate), chronically offered to the animals. Male rats ate more chocolate than females, and they also exhibited a higher weight gain, abdominal fat deposition, and higher plasma levels of total cholesterol, LDL-cholesterol and glucose. The stress exposure decreased body weight, increased adrenal weight and decreased plasma insulin levels. Overall, female rats had lower plasma insulin levels and chocolate consumption prevented the increased adrenal gland weight after exposure to chronic stress, suggesting a reduction of stress effects induced by palatable food consumption. Taken together, these results suggest a peculiar metabolic pattern, related to energy store and expenditure, in stressed animals receiving a palatable diet. Since these effects were sex-specific, we may also propose that females and males subjected to restraint stress and chocolate consumption are differentially affected.


Mobile-based effective vaccine management tool: An m-health initiative implemented by UNICEF in Bihar.
UNICEF launched the mobile-based Effective Vaccine Management (EVM) system in Bihar in 2014 along with the state government to electronically capture information and identify gaps in the existing vaccine management system for appropriate action. This study accessed the implementation process of this initiative. Quantitative data related to vaccine supply chain management indicators were collected in November-December 2015 using factsheets and dashboards, representing the situation of the vaccine supply and cold chain management system at regular intervals since the launch. In-depth interviews were conducted with the program specialists to understand the initiative's genesis, its challenges and strengths. This initiative resulted in an increased cold chain space from 49% (July 2014) to 87% (September 2015), deployment of sufficient human resource; 38 cold chain technicians for regular maintenance of the machines and equipment, installation of necessary equipment, and upgradation of state and regional vaccine stores. In health facilities, district vaccine stores, and regional vaccine stores, marked improvements were observed in the overall EVM criteria indicators (82%, 84%, and 80% in September 2015, respectively, as against 51%, 46%, and 43% in July 2014, respectively) as well as EVM category indicators (83%, 84%, and 76% in September 2015, respectively, as compared to 54%, 53%, and 54% in July 2014, respectively). The EVM mobile initiative was successfully implemented and it complies with its objective of providing experienced guidance to the human resource responsible for vaccine cold chain management. The initiative is scalable and its sustainability depends on its thoughtful merger with the existing immunization ecosystem.


Transforming governance or reinforcing hierarchies and competition: examining the public and hidden transcripts of the Global Fund and HIV in India.
Global health initiatives (GHIs) have gained prominence as innovative and effective policy mechanisms to tackle global health priorities. More recent literature reveals governance-related challenges and their unintended health system effects. Much less attention is received by the relationship between these mechanisms, the ideas that underpin them and the country-level practices they generate. The Global Fund has leveraged significant funding and taken a lead in harmonizing disparate efforts to control HIV/AIDS. Its growing influence in recipient countries makes it a useful case to examine this relationship and evaluate the extent to which the dominant public discourse on Global Fund departs from the hidden resistances and conflicts in its operation. Drawing on insights from ethnographic fieldwork and 70 interviews with multiple stakeholders, this article aims to better understand and reveal the public and the hidden transcript of the Global Fund and its activities in India. We argue that while its public transcript abdicates its role in country-level operations, a critical ethnographic examination of the organization and governance of the Fund in India reveals a contrasting scenario. Its organizing principles prompt diverse actors with conflicting agendas to come together in response to the availability of funds. Multiple and discrete projects emerge, each leveraging control and resources and acting as conduits of power. We examine how management of HIV is punctuated with conflicts of power and interests in a competitive environment set off by the Fund protocol and discuss its system-wide effects. The findings also underscore the need for similar ethnographic research on the financing and policy-making architecture of GHIs.


Phototropism: translating light into directional growth.
Phototropism allows plants to align their photosynthetic tissues with incoming light. The direction of incident light is sensed by the phototropin family of blue light photoreceptors (phot1 and phot2 in Arabidopsis), which are light-activated protein kinases. The kinase activity of phototropins and phosphorylation of residues in the activation loop of their kinase domains are essential for the phototropic response. These initial steps trigger the formation of the auxin gradient across the hypocotyl that leads to asymmetric growth. The molecular events between photoreceptor activation and the growth response are only starting to be elucidated. In this review, we discuss the major steps leading from light perception to directional growth concentrating on Arabidopsis. In addition, we highlight links that connect these different steps enabling the phototropic response.


Targeted disruption of the mouse transforming growth factor-beta 1 gene results in multifocal inflammatory disease.
Transforming growth factor-beta 1 (TGF-beta 1) is a multifunctional growth factor that has profound regulatory effects on many developmental and physiological processes. Disruption of the TGF-beta 1 gene by homologous recombination in murine embryonic stem cells enables mice to be generated that carry the disrupted allele. Animals homozygous for the mutated TGF-beta 1 allele show no gross developmental abnormalities, but about 20 days after birth they succumb to a wasting syndrome accompanied by a multifocal, mixed inflammatory cell response and tissue necrosis, leading to organ failure and death. TGF-beta 1-deficient mice may be valuable models for human immune and inflammatory disorders, including autoimmune diseases, transplant rejection and graft versus host reactions.


Rehabilitation outcomes of stroke patients treated with multi-modal endovascular reperfusion therapy.
The aim of this study was to investigate the influence of multi-modal endovascular reperfusion therapy (MMRT) on functional outcomes following rehabilitation. Data from 14 MMRT-treated patients were analyzed and compared to MMRT-ineligible, age and stroke severity-matched patients treated at the same Neurological and Rehabilitation departments. Neurological evaluation was assessed with the NIH stroke scale (NIHSS). Activity of daily living was measured using the FIMTM instrument. Functional outcome was measured using the modified Rankin scale (mRS). The baseline characteristics of both groups were similar. NIHSS scores were lower in the MMRT group and they had slightly better functional and rehabilitation scores on admission to rehabilitation. At the end of rehabilitation, more MMRT-treated patients reached functional independence (mRS≤2; 50% vs. 7% respectively P=0.03). FIM scores were also higher in the MMRT group (mean score 93.3 vs. 87.7, respectively) but the difference did not reach significance. The delta in FIM and NIHSS scores obtained during rehabilitation did not significantly differ between the groups. MMRT remained a significant modifier of good outcome after regression analysis (OR 21.5 95% CI 1.1-410). MMRT-treated patients have better chances of attaining independence after rehabilitation therapy. However, the additional improvements gained while in active rehabilitation were independent of reperfusion status.


The neuroendocrine protein VGF is sorted into dense-core granules and is secreted apically by polarized rat thyroid epithelial cells.
We have expressed the neuroendocrine VGF protein in FRT rat thyroid cells to study the molecular mechanisms of its sorting to the regulated and polarized pathways of secretion. By immunoelectron microscopy, we have demonstrated that VGF localizes in dense-core granules. Rapid secretion of VGF is induced by PMA stimulation. Moreover, human chromogranin B, a protein of the regulated pathway, co-localizes in the same granules with VGF. In confluent, FRT monolayers on filters protein secretion occur from the apical cell domain. VGF deletion mutants have been generated. By confocal microscopy, we have found that in transient transfection, all mutant proteins are sorted into granules and co-localize with the full-length VGF. They all retain the apical polarity of secretion. We also found that intracellular VGF and its deletion mutants are largely in an aggregated form. We conclude that FRT thyroid cells correctly decode the sorting information of VGF. The signals present on the protein to enter the granules and to be secreted apically cannot be separated from each other and are not in just one discrete portion of the protein. We propose that selective aggregation might represent the signal for sorting VGF to the regulated, apical route.


Cdc5 interacts with the Wee1 kinase in budding yeast.
Development of a multicellular organism requires that mitosis and morphogenesis be coordinated. These processes must also be synchronized during the growth of unicellular organisms. In the yeast Saccharomyces cerevisiae, mitosis is dependent on the prior growth of a daughter cell in the form of a bud. Overexpression of wild-type Polo-like kinase Cdc5 or a catalytically inactive form resulted in the formation of multinucleate cells in budding yeast. Immunofluorescence analysis of these multinulceate cells showed that mitosis and bud formation were no longer linked. Others have shown that Swe1 is required for coupling mitosis to bud formation during a perturbed cell cycle. When the normal pathway of bud formation is perturbed, Swe1 functions to delay mitosis through negative regulation of Clb/Cdk. In cells lacking Swe1, multinucleate cells are formed in response to delays in bud formation. Affinity purification, two-hybrid analysis, and mutant characterization results suggested that Cdc5 and Swe1 interact. From these results, we conclude that multinucleate formation in response to Cdc5 overexpression is linked to titration of Swe1 function. These results also suggest that Cdc5 may be a negative regulator of Swe1.


Investigation of the origin of selectivity in cavitand-based supramolecular sensors.
The sensing properties of functionalized cavitands have been studied by thin-film coating TMSR chemical sensors and by measuring their responses towards model analytes. We studied the relationship between the sensor performance, in terms of sensitivity and selectivity, and the molecular recognition properties of the cavitands. The Langmuir-like shape of the adsorption isotherm, obtained in the case of short-chain alcohols, demonstrated that selective binding can be achieved by the synergistic interactions of the cavity and the bridging PO(in) groups. In the absence of these substituents, the peripheral alkyl chains necessary for the formation of highly permeable thin films attenuate the cavity effect because of nonspecific dispersion interactions. This completely overrides the response originating from molecular recognition. The same effect is observed when the PO groups are oriented outward from the cavity. The use of multivariate chemometrics and the study of the correlations between sensors sensitivity and analyte properties provided further evidence of molecular recognition phenomena, whose intensity is enhanced by the permanent free volume created by the rigid cavity surrounding the PO(in) group.


Cognitive functioning transitions, health expectancies, and inequalities among elderly people in China: A nationwide longitudinal study.
In China, cognitive impairment has become a huge challenge owing to the rapid aging process. We investigate cognitive health expectancy and potential factors leading to inequalities of cognitive health for Chinese older people. The study included 19 943 participants aged 65 to 95 at the first observation in Chinese Longitudinal Healthy Longevity Survey collected during 2002-2014. Cognitive impairment was categorized into none, mild, and severe by the score of cMMSE. Multistate models based on continuous-time Markov process were applied to calculate cognitive health expectancies and estimate hazard ratio from no impairment to impairment for potential factors. Urban women and men aged 65 were expected to spend 31.18% and 19.82% of their remaining 17.46 and 15.45 years with cognitive impairment; meanwhile, rural women and men aged 65 were expected to spend 35.31% and 21.39% of their remaining 16.73 and 14.87 years with cognitive impairment. Women achieving lower educational attainment (HR1-6 years = 0.78, 95% CI, 0.71-0.87; HR7+ years = 0.59, 95% CI, 0.49-0.70) than men and rural residents having less access to medical service (HR = 0.79, 95% CI, 0.68-0.92) and doing less regular exercise (HR = 0.87, 95% CI, 0.80-0.96) than urban people caused the differences of cognitive health for genders and regions. Women and rural people experience less cognitive health expectancies compared with their counterparts, respectively. Chinese government should pay more attention to rural women and make efforts to reduce the inequalities of cognitive health by increasing opportunities of education for women and improving access to medical service and healthy lifestyle for rural people.


Lipid composition and biosynthesis in the gingiva of the domestic cat.
Periodontal disease in the domestic cat may assume debilitating and serious consequences; however, little is known of the biochemical composition or metabolism of feline gingiva in health or disease. In this report the chemical composition and metabolism of gingival lipids from inflamed an non-inflamed sites is presented and compared to other species with naturally occurring periodontitis. The neutral and phospholipid composition of feline gingiva was found to be distinct from that of porcine and human. As a measure of de novo lipid synthesis, the total incorporation of 14C-acetate into fractionated lipid components was determined and revealed an approximate 2 to 3 fold decrease in inflamed versus non-inflamed gingiva. The decrease in 14C-acetate incorporation appeared to result from a 2-fold increase in free acetate pools in inflamed compared to non-inflamed gingival samples, since total lipase and phospholipase activity were comparable in inflamed and non-inflamed gingiva and total lipid composition was not changed between inflamed and non-inflamed sites. These data are similar to those reported for periodontally involved human gingival tissue and suggest a common effect of periodontal inflammation on lipid metabolism in both species.


A suspended core nanofiber with unprecedented large diameter ratio of holey region to core.
For a suspended core nanofiber, the holey region is expected to be as large as possible to propagate the light at wavelengths as long as possible. Additionally, a large holey region is significant for its applications in sensors. However, the fabrication of nanofiber with large holey region is still a challenge so far. In this paper a method, which involves pumping positive pressure of nitrogen gas in both the cane fabrication and fiber-drawing processes, was proposed. A suspended core nanofiber, with a core diameter of around 480 nm and an unprecedented diameter ratio of holey region to core (DRHC) of at least 62, was fabricated in the length of several hundred meters. Owing to the large holey region, the confinement loss of the suspended core nanofiber is insignificant when the wavelength of light propagated in it is 1700 nm. For this fabrication technique, the nanowire length, fabrication efficiency, and the uniformity in the diameter are much superior to those of the nanowires fabricated in other ways. Finally, single mode third harmonic generation was observed by this nanofiber under the pump of a 1557 nm femtosecond fiber laser. This work shows the prospect of fabrication of nanostructured waveguide in glass materials by an inflation technique.


Glycosylation Effects on FSH-FSHR Interaction Dynamics: A Case Study of Different FSH Glycoforms by Molecular Dynamics Simulations.
The gonadotropin known as follicle-stimulating hormone (FSH) plays a key role in regulating reproductive processes. Physiologically active FSH is a glycoprotein that can accommodate glycans on up to four asparagine residues, including two sites in the FSHα subunit that are critical for biochemical function, plus two sites in the β subunit, whose differential glycosylation states appear to correspond to physiologically distinct functions. Some degree of FSHβ hypo-glycosylation seems to confer advantages toward reproductive fertility of child-bearing females. In order to identify possible mechanistic underpinnings for this physiological difference we have pursued computationally intensive molecular dynamics simulations on complexes between the high affinity site of the gonadal FSH receptor (FSHR) and several FSH glycoforms including fully-glycosylated (FSH24), hypo-glycosylated (e.g., FSH15), and completely deglycosylated FSH (dgFSH). These simulations suggest that deviations in FSH/FSHR binding profile as a function of glycosylation state are modest when FSH is adorned with only small glycans, such as single N-acetylglucosamine residues. However, substantial qualitative differences emerge between FSH15 and FSH24 when FSH is decorated with a much larger, tetra-antennary glycan. Specifically, the FSHR complex with hypo-glycosylated FSH15 is observed to undergo a significant conformational shift after 5-10 ns of simulation, indicating that FSH15 has greater conformational flexibility than FSH24 which may explain the more favorable FSH15 kinetic profile. FSH15 also exhibits a stronger binding free energy, due in large part to formation of closer and more persistent salt-bridges with FSHR.


Polymorphisms of the P-selectin gene and risk of myocardial infarction in men and women in the ECTIM extension study. Etude cas-temoin de l'infarctus myocarde.
Studies in animal models and humans implicate cell adhesion molecules in atherogenesis but their role in mediating the risk of myocardial infarction is unclear. The ECTIM (étude cas-temoin de l'infarctus myocarde) extension study was established to determine whether a previously implicated polymorphism of the P-selectin gene was associated with myocardial infarction risk in men and women in Belfast and Glasgow. PATIENTS AND STUDY SETTING: 696 cases with a recent myocardial infarction and 561 age matched controls (both male and female) were recruited into a case-control study in MONICA project areas of Belfast and Glasgow. Demographic and lifestyle information was collected by interview administered questionnaire, and each subject was examined and provided a blood sample for DNA extraction. The polymerase chain reaction (PCR) was used to amplify regions encompassing the P-selectin Thr-->Pro (A/C) polymorphism at position 715. Genotype odds ratios for myocardial infarction were estimated by logistic regression adjusted for population, age, and sex. There was no significant association between conventional risk factors (such as hypercholesterolaemia, increased body mass index, or raised blood pressure) and either the rare or the common Pro(715) allele of the P-selectin gene in controls. Overall, comparing Pro(715)/Pro(715) and Pro(715)/Thr(715) with Thr(715)/Thr(715), with adjustment for centre, age, and sex, the odds ratio was 0.78 (95% confidence interval 0.60 to 1.00) (p = 0.054), indicating a "protective" effect of the less common Pro(715) allele. There was no significant heterogeneity in odds ratios between men and women either in this sample or when combined with the original ECTIM subjects. In a large population based study in two regions of the UK, we have been able to corroborate the earlier ECTIM findings of a lower frequency of the Thr/Pro(715) polymorphism in subjects with myocardial infarction. An apparently "protective effect" of similar magnitude also seems to apply to women.


A rapid and precise method for the determination of urea.
A method is described for the determination of urea in plasma and urine. The effects of variations in the experimental conditions are examined and the results of recovery experiments and other tests of precision and accuracy are reported. In comparison with other methods in current use, this method has distinct advantages in sensitivity, simplicity, and precision, thus economizing in time, sample volume, reagents, and equipment.


Epidermal growth factor receptor in vulvar malignancies and its relationship to metastasis and patient survival.
To evaluate the level of epidermal growth factor receptor (EGF-R) expression in vulvar malignancies and to determine if a correlation exists between EGF-R levels and metastasis or patient survival. All patients with a diagnosis of invasive squamous cell carcinoma of the vulva who were treated at our institution with a primary radical vulvectomy and inguinal lymph node dissection from 1983 to 1993 were eligible for the study. Sixty-one patients with available tissue blocks of benign vulvar epithelium, the primary malignant vulvar lesion, and groin node metastasis (when positive) were included in the study. Semiquantitative EGF-R expression was determined in a blinded fashion utilizing immunohistochemical staining of appropriate tissue samples. Survival was calculated utilizing Kaplan-Meier life table analysis based upon disease-free survival. A significant increase (P < 0.001) in mean EGF-R levels was demonstrated in the primary tumor (67%) versus benign vulvar epithelium (31%). In the 14 patients with lymph node metastasis, the mean EGF-R level in the primary tumor was 65% versus 88% in the metastatic lesion (P < 0.001). The likelihood of lymph node metastasis was elevated in those patients with a benign tissue EGF-R level > or =40% (P < 0.03) and in those patients with a primary tumor EGF-R level > or =90% (P < 0.025). Life table analysis revealed a cumulative disease-free survival of 45% for all patients. Disease-free survival in those patients with EGF-R levels > or =90% in the primary tumor was 25%, contrasting with a disease-free survival of 54% in those patients with EGF-R levels <90% (P < 0.05). There is a progressive increase in EGF-R expression from benign vulvar epithelium to primary malignant tissue to metastatic lesions within the same patient. Increased expression of EGF-R in the primary vulvar malignancy is significantly associated with lymph node metastasis and decreased patient survival. Increased expression of EGF-R in histologically benign vulvar epithelium has a significant association with lymph node metastasis and may predict decreased patient survival.


Breakage is the norm: use of condoms and lubrication in anal sex among Black South African men who have sex with men.
This paper explores condom use and lubrication practices among Black men who have sex with men in South African townships. Results are from 81 in-depth individual interviews conducted among a purposive sample from four townships surrounding Pretoria as part of a larger qualitative study. Awareness that condoms should be used to have safer anal sex was ubiquitous. Fewer men reported that lubricants should be used to facilitate anal intercourse. Partner pressure and partner distrust were the most common barriers cited for not using condoms and lubricants. Knowledge about condom-lubricant compatibility was rare. Condom problems were a norm, with widespread expectations of condom failure. Men's subjectivities - their perceptions of and preferences for specific brands, types and flavours of condoms and lubricants - influenced engagement with such safer-sex technologies. However, what was available in these settings was often neither what men needed nor preferred. Findings show the need to enhance access to appropriate and comprehensive: safer-sex supplies, health services and health education, and underline the importance of efforts to develop targeted programmes relevant to experiences of men who have sex with men in the South African context.


Consumer Preferences and Socioeconomic Factors Decided on Plantain and Plantain-Based Products in the Central Region of Cameroon and Oyo State, Nigeria.
Plantain is a key staple food in Central and West Africa, but there is limited understanding of its market in Africa. In addition, the cooking methods for enhancing the nutritional value, consumer preference, and willingness to pay for plantain and plantain-based products are not well understood. The knowledge gaps in the market and consumer dimension of the food chain need to be known to increase plantain utilization and guide breeding efforts. This research contributes by examining the cooking methods, consumer preference, and willingness to pay for plantain and plantain-based products in Cameroon and Nigeria. A household survey sample of 454 Cameroonian consumers in four divisions of Central Region and 418 Nigerian consumers in seven government areas of Oyo State in southwest Nigeria was the basis for the analysis. The results showed some levels of similarity and difference in the consumption and cooking of boiled, roasted, and fried plantain in both countries. The trend in consumption of all plantain-based products was constant in Cameroon but increased in Nigeria. The most important factor influencing Cameroonian consumers' choice of plantain and its products was taste, while the nutrition trait influenced Nigerian consumers. Both Cameroonian and Nigerian consumers considered packaging, location of produce, and size and quantity as the least important factors. In addition, socioeconomic characteristics were significant determinants of consumers' choices to consume plantain and its products. Gender significantly influenced (p < 0.05) taste, while nutrition was significantly driven (p < 0.05) by education and annual income. Household size played a significant role (p < 0.05) in consumers' choices when the price was considered. These findings serve as a guideline to improve existing products to match the needs of consumers in each country and develop products for different consumer segments and potentially increase production.


Disturbances of the Perioperative Microbiome Across Multiple Body Sites in Patients Undergoing Pancreaticoduodenectomy.
The goals of this study were to characterize bacterial communities within fecal samples, pancreatic fluid, bile, and jejunal contents from patients undergoing pancreaticoduodenectomy (PD) and to identify associations between microbiome profiles and clinical variables. Fluid was collected from the pancreas, common bile duct, and proximal jejunum from 50 PD patients. Postoperative fecal samples were also collected. The microbial burden within samples was quantified with droplet digital polymerase chain reaction. Bacterial 16S ribosomal RNA gene sequences were amplified, sequenced, and analyzed. Data from fecal samples were compared with publicly available data obtained from volunteers. Droplet digital polymerase chain reaction confirmed the presence of bacteria in all sample types, including pancreatic fluid. Relative to samples from the American Gut Project, fecal samples from PD patients were enriched with Klebsiella and Bacteroides and were depleted of anaerobic taxa (eg, Roseburia and Faecalibacterium). Similar patterns were observed within PD pancreas, bile, and jejunal samples. Postoperative fecal samples from patients with a pancreatic fistula contained increased abundance of Klebsiella and decreased abundance of commensal anaerobes, for example, Ruminococcus. This study confirms the presence of altered bacterial populations within samples from PD patients. Future research must validate these findings and may evaluate targeted microbiome modifications to improve outcomes in PD patients.


Assessment of regional variation in streamflow responses to urbanization and the persistence of physiography.
Aquatic ecosystems are sensitive to the modification of hydrologic regimes, experiencing declines in stream health as the streamflow regime is altered during urbanization. This study uses streamflow records to quantify the type and magnitude of hydrologic changes across urbanization gradients in nine U.S. cities (Atlanta, GA, Baltimore, MD, Boston, MA, Detroit, MI, Raleigh, NC, St. Paul, MN, Pittsburgh, PA, Phoenix, AZ, and Portland, OR) in two physiographic settings. Results indicate similar development trajectories among urbanization gradients, but heterogeneity in the type and magnitude of hydrologic responses to this apparently uniform urban pattern. Similar urban patterns did not confer similar hydrologic function. Study watersheds in landscapes with level slopes and high soil permeability had less frequent high-flow events, longer high-flow durations, lower flashiness response, and lower flow maxima compared to similarly developed watersheds in landscape with steep slopes and low soil permeability. Our results suggest that physical characteristics associated with level topography and high water-storage capacity buffer the severity of hydrologic changes associated with urbanization. Urbanization overlain upon a diverse set of physical templates creates multiple pathways toward hydrologic impairment; therefore, we caution against the use of the urban homogenization framework in examining geophysically dominated processes.


Overview of magnetic resonance imaging for management of relapsing-remitting multiple sclerosis in everyday practice.
Although the value of magnetic resonance imaging (MRI) for diagnosis/differential diagnosis of patients with clinically isolated syndromes suggestive of multiple sclerosis (MS) is widely accepted, adoption of MRI into clinical practice to monitor disease evolution remains a work in progress. However, an accumulating body of evidence points to a central role for MRI in managing patients with relapsing-remitting MS along the disease continuum. Routine MRI surveillance provides insight into disease activity that is not evident clinically and this information, in turn, can be used to inform prognosis and guide treatment decisions. In Europe, practical guidelines have been developed to reduce the heterogeneity of imaging (both intracentre and intercentre) and improve the quality of MRI assessment and interpretation. Aimed at the general neurologist, this review explores some of the issues associated with MRI and examines evidence supporting its use for routine monitoring of MS patients in everyday practice.


Analysis of the Clinicopathological Characteristics of Gastric Cancer in Extremely Old Patients.
Gastric cancer is the third-leading cause of cancer-related death in Korea. As the Korean population is ageing, the number of extremely old patients with this disease is increasing. This study examined the clinicopathological characteristics of gastric cancer in extremely old (over 85 years) patients who received treatment or conservative observations and compared the treatment outcomes according to the treatment modality. A total of 170 patients over 85 years of age were diagnosed with gastric cancer. Of these, 81 underwent treatment for gastric cancer and 89 received conservative observations. The clinicopathological characteristics of the treatment and conservative groupswere compared. The mean age of the patients was 86.5 years. The conservative group included significantly more patients with older ages, macroscopically advanced cancer and upper-middle located cancer. The overall survival rate of the treatment group was significantly higher than that of the conservative group. The disease-specific mortality rate was significantly lower in the treatment group than in the conservative group. Multivariate analysis revealed the clinical course, alarm sign, and macroscopic classification to be independent prognosis factors. By itself, the chronological age should not be used as a strategy to determine whether treatmentwill be administered for gastric cancer. Patients who have early gastric cancer or lower-risk preexisting comorbidities should not be discouraged from treatment, even if they are older than 85 years.


Dosimetric uncertainties of three-dimensional dose reconstruction from two-dimensional data in a multi-institutional study.
Inconsistencies in the treatment planning process leading to dosimetric uncertainties may affect conclusions drawn from interinstitutional radiation oncology clinical trials. The purpose of this study was to assess the dosimetric uncertainties resulting from the process of reconstructing three-dimensional dose distributions from two-dimensional treatment plan information provided by participating institutions in a randomized clinical trial. This study was based on American College of Radiology Protocol #427, Locally Advanced Multi-Modality Protocol; a multi-institutional phase II randomized study involving radiation therapy for patients with inoperable non-small cell lung cancer. Several sources of dosimetric uncertainty were identified and analyzed, including image quality of hard-copy computed tomography (CT) images, slice spacing of CT scans, treatment position, interpretations of target volumes by radiation oncologists, the contouring of normal anatomic structures, and the use of common beam models for all dose calculations. Each source of uncertainty was investigated using a set of plans, with the ideal characteristics of digital images with 3-mm axial slice spacing and a flat couch, consisting of eight cases from Vanderbilt University Medical Center with electronically transferred CT data. The target volume DVH values were dependent on the additional uncertainty introduced by differences in delineation of the target volumes by the participating radiation oncologists. The DVH values for the lungs and heart were dependent on image quality and treatment position. Esophagus DVH values were not dependent on any of the sources of uncertainty. None of the structure DVH values were dependent on slice thickness or variations in the contouring of normal anatomic structures. Reconstruction of three-dimensional dose distributions from two-dimensional treatment plan information may be useful in cases for which digital CT data is not available or for historical data review. However, dosimetric accuracy will depend on image quality of the treatment planning CT data and consistency in the delineation of tumor volumes.


Determinants of rural women's health: a qualitative study in Southwest Ontario.
The influences of gender and geography are increasingly being acknowledged as central to a comprehensive understanding of health. Since little research on rural women's health has been conducted, an in-depth qualitative approach is necessary to gain a better initial understanding of this population. To explore the determinants of health and their influence on rural women's health. From November 2004 to September 2005, 9 focus groups and 3 individual interviews were conducted in 7 rural southwestern Ontario communities. Sixty-five rural residents aged 26 years and older participated in the study. Semi-structured interview questions were used to elicit participants' perceptions regarding determinants of rural women's health. Four Health Canada determinants (employment, gender, health services, and social environments) and 3 new determinants (rural change, rural culture, and rural pride) emerged as key to rural women's health. Although health determinants affect both urban and rural people, this qualitative study revealed that rural women experience health determinants in unique ways and that rural residents may indeed have determinants of their health that are particular to them. More research is needed to explore the nature and effects of determinants of health for rural residents in general, and rural women in particular.


Providing Trainees With an Introduction and Decision-Making Framework for Pursuing an Academic Residency Position.
Medical students are a significant source of the future academic medicine workforce, but surprisingly little is typically done to promote academic medicine career awareness during the early stages of medical education. To raise awareness among medical students of the critical role an academic residency plays in pursuing a future academic career, we developed a workshop that describes what an academic residency entails and what to consider when selecting such a program. The six-step Kern model was applied to develop a multimodal workshop to help trainees understand characteristics of and explore their interest in an academic residency program. The 60-minute workshop consists of a PowerPoint presentation, reflection exercise, and case scenarios to help trainees learn and apply new skills in selecting a residency program. The workshop was offered to diverse medical students at nine different medical schools across the country between July 2016 and May 2017. Of the 115 workshop participants who completed the evaluation form, over 95% agreed or strongly agreed that each of the learning objectives was met. A paired t test of learner responses pre- and postworkshop demonstrated significantly increased confidence in finding an academic residency position. This workshop fills an important gap in raising medical students' awareness of how to pursue an academic medicine career by highlighting the critical role of an academic residency program. Participants liked the use of vignettes and small-group discussion to better prepare them in exploring and benefiting from an academic residency position.


Dual-wavelength low-coherence instantaneous phase-shifting interferometer to measure the shape of a segmented mirror with subnanometer precision.
We present a noncontact optical metrology measuring the pistons and tip/tilt angles of the 61 hexagonal segments of a compact-sized segmented mirror. The instrument has been developed within the scope of a design study for the European Extremely Large Telescope (E-ELT). It is used as reference sensor for cophasing of the mirror segments in closed-loop control. The mirror shape is also measured by different types of stellar light-based phasing cameras whose performances will be evaluated with regard to a future E-ELT. Following a description of the system architecture, the second part of the paper presents experimental results demonstrating the achieved precision: 0.48 nm rms in piston and 74 nrad rms in tip/tilt.


miniMDS: 3D structural inference from high-resolution Hi-C data.
Recent experiments have provided Hi-C data at resolution as high as 1 kbp. However, 3D structural inference from high-resolution Hi-C datasets is often computationally unfeasible using existing methods. We have developed miniMDS, an approximation of multidimensional scaling (MDS) that partitions a Hi-C dataset, performs high-resolution MDS separately on each partition, and then reassembles the partitions using low-resolution MDS. miniMDS is faster, more accurate, and uses less memory than existing methods for inferring the human genome at high resolution (10 kbp). A Python implementation of miniMDS is available on GitHub: https://github.com/seqcode/miniMDS . email@example.com. Supplementary data are available at Bioinformatics online.


Research Opportunities in the Area of People With Autism Spectrum Disorder.
The American Occupational Therapy Association (AOTA) Evidence-Based Practice Project has developed a table summarizing the research opportunities on people with autism spectrum disorder. The table provides an overview of the state of the current available evidence on interventions within the scope of occupational therapy practice and is based on the systematic reviews from the AOTA Evidence-Based Practice Guidelines Series. Researchers, students, and clinicians can use this information in developing innovative research to answer important questions within the occupational therapy field.


Effects of jasmonic acid, ethylene, and salicylic acid signaling on the rhizosphere bacterial community of Arabidopsis thaliana.
Systemically induced resistance is a promising strategy to control plant diseases, as it affects numerous pathogens. However, since induced resistance reduces one or both growth and activity of plant pathogens, the indigenous microflora may also be affected by an enhanced defensive state of the plant. The aim of this study was to elucidate how much the bacterial rhizosphere microflora of Arabidopsis is affected by induced systemic resistance (ISR) or systemic acquired resistance (SAR). Therefore, the bacterial microflora of wild-type plants and plants affected in their defense signaling was compared. Additionally, ISR was induced by application of methyl jasmonate and SAR by treatment with salicylic acid or benzothiadiazole. As a comparative model, we also used wild type and ethylene-insensitive tobacco. Some of the Arabidopsis genotypes affected in defense signaling showed altered numbers of culturable bacteria in their rhizospheres; however, effects were dependent on soil type. Effects of plant genotype on rhizosphere bacterial community structure could not be related to plant defense because chemical activation of ISR or SAR had no significant effects on density and structure of the rhizosphere bacterial community. These findings support the notion that control of plant diseases by elicitation of systemic resistance will not significantly affect the resident soil bacterial microflora.


SvSXP: a Strongylus vulgaris antigen with potential for prepatent diagnosis.
Strongyle parasites are ubiquitous in grazing horses. Strongylus vulgaris, the most pathogenic of the large strongyles, is known for its extensive migration in the mesenteric arterial system. The lifecycle of S. vulgaris is characterised by a long prepatent period where the migrating larvae are virtually undetectable as there currently is no test available for diagnosing prepatent S. vulgaris infection. Presence of S. vulgaris larvae in the arterial system causes endarteritis and thrombosis with a risk of non-strangulating intestinal infarctions. Emergence of anthelmintic resistance among cyathostomins has led to recommendations of reduced treatment intensity by targeting horses that exceed a predetermined strongyle faecal egg count threshold. One study suggests an apparent increase in prevalence of S. vulgaris on farms where reduced anthelmintic treatment intensity has been implemented. These issues highlight the need for an accurate and reliable assay for diagnosing prepatent S. vulgaris infection. Immunoscreening of a larval S. vulgaris cDNA library using hyperimmune serum raised against S. vulgaris excretory/secretory antigens was performed to identify potential diagnostic antigens. Immunoreactive clones were sequenced, one potential antigen was characterised, expressed as a recombinant protein, initially evaluated by western blot (WB) analysis, the diagnostic potential of the IgG subclasses was evaluated by ELISA, and the diagnostic accuracy evaluated using serum from 102 horses with known S. vulgaris infection status. The clone expressing the potential antigen encoded a S. vulgaris SXP/RAL2 homologue. The recombinant protein, rSvSXP, was shown to be a potential diagnostic antigen by WB analysis, and a target of serum IgGa, IgG(T) and total IgG in naturally infected horses, with IgG(T) antibodies being the most reliable indicator of S. vulgaris infection in horses. Evaluation of diagnostic accuracy of the ELISA resulted in a sensitivity of 73.3%, a specificity of 81.0%, a diagnostic odds ratio of 11.69; a positive likelihood ratio (LR) of 3.85 and a negative LR was 0.33. The area under the ROC curve was 0.820. IgG(T) antibodies to recombinant SvSXP show potential for use as an antigen for prepatent diagnosis of migrating stages of S. vulgaris with moderate to good diagnostic accuracy.


A nonparametric Bayesian framework for constructing flexible feature representations.
Representations are a key explanatory device used by cognitive psychologists to account for human behavior. Understanding the effects of context and experience on the representations people use is essential, because if two people encode the same stimulus using different representations, their response to that stimulus may be different. We present a computational framework that can be used to define models that flexibly construct feature representations (where by a feature we mean a part of the image of an object) for a set of observed objects, based on nonparametric Bayesian statistics. Austerweil and Griffiths (2011) presented an initial model constructed in this framework that captures how the distribution of parts affects the features people use to represent a set of objects. We build on this work in three ways. First, although people use features that can be transformed on each observation (e.g., translate on the retinal image), many existing feature learning models can only recognize features that are not transformed (occur identically each time). Consequently, we extend the initial model to infer features that are invariant over a set of transformations, and learn different structures of dependence between feature transformations. Second, we compare two possible methods for capturing the manner that categorization affects feature representations. Finally, we present a model that learns features incrementally, capturing an effect of the order of object presentation on the features people learn. We conclude by considering the implications and limitations of our empirical and theoretical results.


Modeling daily and weekly moderate and vigorous physical activity using zero-inflated mixture Poisson distribution.
Recently developed accelerometer devices have been used in large epidemiological studies for continuous and objective monitoring of physical activities. Typically, physical movements are summarized as minutes in light, moderate, and vigorous physical activities in each wearing day. Because of preponderance of zeros, zero-inflated distributions have been used for modeling the daily moderate or higher levels of physical activity. Yet, these models do not fully account for variations in daily physical activity and cannot be extended to model weekly physical activity explicitly, while the weekly physical activity is considered as an indicator for a subject's average level of physical activity. To overcome these limitations, we propose to use a zero-inflated Poisson mixture distribution that can model daily and weekly physical activity in same family of mixture distributions. Under this method, the likelihood of an inactive day and the amount of exercise in an active day are simultaneously modeled by a joint random effects model to incorporate heterogeneity across participants. If needed, the method has the flexibility to include an additional random effect to address extra variations in daily physical activity. Maximum likelihood estimation can be obtained through Gaussian quadrature technique, which is implemented conveniently in an R package GLMMadaptive. Method performances are examined using simulation studies. The method is applied to data from the Hispanic Community Health Study/Study of Latinos to examine the relationship between physical activity and BMI groups and within a participant the difference in physical activity between weekends and weekdays.


Access to green space, physical activity and mental health: a twin study.
Increasing global urbanisation has resulted in a greater proportion of the world's population becoming exposed to risk factors unique to urban areas, and understanding these effects on public health is essential. The aim of this study was to examine the association between access to green space and mental health among adult twin pairs. We used a multilevel random intercept model of same-sex twin pairs (4338 individuals) from the community-based University of Washington Twin Registry to analyse the association between access to green space, as measured by the Normalised Difference Vegetation Index and self-reported depression, stress, and anxiety. The main parameter of interest was the within-pair effect for identical (monozygotic, MZ) twins because it was not subject to confounding by genetic or shared childhood environment factors. Models were adjusted for income, physical activity, neighbourhood deprivation and population density. When treating twins as individuals and not as members of a twin pair, green space was significantly inversely associated with each mental health outcome. The association with depression remained significant in the within-pair MZ univariate and adjusted models; however, there was no within-pair MZ effect for stress or anxiety among the models adjusted for income and physical activity. These results suggest that greater access to green space is associated with less depression, but provide less evidence for effects on stress or anxiety. Understanding the mechanisms linking neighbourhood characteristics to mental health has important public health implications. Future studies should combine twin designs and longitudinal data to strengthen causal inference.


Local administration of sarizotan into the subthalamic nucleus attenuates levodopa-induced dyskinesias in 6-OHDA-lesioned rats.
Dyskinesia affects the majority of levodopa-treated parkinsonian patients within 5-10 years of treatment with levodopa. Clinical and preclinical observations suggest that an increase in serotoninergic transmission can contribute to the appearance of dyskinesias. It is thus conceivable that a modulation of synaptic dopamine (DA) levels induced by the inhibition of serotonin (5-HT) release, as a consequence of 5-HT(1A) agonists administration, might alleviate dyskinesias. Since 5-HT(1A) receptors are expressed in the subthalamic nucleus (STN), the aim of the present study was to assess the effect of the intrasubthalamic administration of sarizotan, a compound with full 5-HT(1A) agonist properties, on levodopa-induced dyskinesias in the 6-hydroxydopamine (6-OHDA) model of parkinsonism. Male Sprague-Dawley rats received a unilateral 6-OHDA administration in the nigrostriatal pathway. A test of apomorphine was performed to evaluate dopamine depletion. One week later, a cannula was implanted in the STN. Animals were treated with levodopa (6 mg/kg, i.p., twice at day) for 22 consecutive days. On day 23, several doses (1 ng, 10 ng, or 1 microg) of sarizotan were administered through the cannula to the STN. The higher doses of sarizotan effectively attenuated all levodopa-induced dyskinesias including axial, limb, and orolingual subtypes. These results suggest that the STN is a target structure for the antidyskinetic action of sarizotan and indicate that drug-mediated modulation of STN activity may be an alternative option for the treatment of levodopa-induced dyskinesias in Parkinson's disease.


Enhanced Platelet-Rich Plasma (ePRP) Stimulates Wound Healing through Effects on Metabolic Reprogramming in Fibroblasts.
As a source of growth factors for expediting wound healing and tissue regeneration, plasma-rich plasma (PRP) has been extensively applied in diverse fields including orthopaedics, ophthalmology, oral and maxillofacial surgery, dentistry, and gynaecology. However, the function of PRP in metabolic regulations remains enigmatic. A standardized method was devised herein to enrich growth factors and to lyophilize it as enhanced PRP (ePRP) powder, which could become ubiquitously available without mechanical centrifugation in clinical practice. To identify metabolic reprogramming in human dermal fibroblasts under ePRP treatment, putative metabolic targets were identified by transcriptome profiling and validated for their metabolic effects and mechanism. ePRP does not only promote wound healing but re-aligns energy metabolism by shifting to glycolysis through stimulation of glycolytic enzyme activity in fibroblasts. On the contrary, oxygen consumption rates and several mitochondrial respiration activities were attenuated in ePRP-treated fibroblasts. Furthermore, ePRP treatment drives the mitochondrial resetting by hindering the mitochondrial biogenesis-related genes and results in a dampened mitochondrial mass. Antioxidant production was further increased by ePRP treatment to prevent reactive oxygen species formation. Besides, ePRP also halts the senescence progression of fibroblasts by activating SIRT1 expression. Importantly, the glycolytic inhibitor 2-DG can completely reverse the ePRP-enhanced wound healing capacity, whereas the mitochondrial inhibitor oligomycin cannot. This is the first study to utilize PRP for comprehensively investigating its effects on the metabolic reprogramming of fibroblasts. These findings indicate that PRP's primary metabolic regulation is to promote metabolic reprogramming toward glycolytic energy metabolism in fibroblasts, preserving redox equilibrium and allowing anabolic pathways necessary for the healing and anti-ageing process.


Anticholinergic burden in older women: not seeing the wood for the trees?
To identify medicines contributing to and describe predictors of anticholinergic burden among community-dwelling older Australian women. Retrospective longitudinal analysis of data from the Australian Longitudinal Study on Women's Health linked to Pharmaceutical Benefits Scheme medicines data from 1 January 2008 to 30 December 2010; for 3694 women born in 1921-1926. Anticholinergic burden calculated from Anticholinergic Drug Scale (ADS) scores derived from ADS levels (0 to 3) for all medicines used by each woman, summed over each 6-month period (semester), medicines commonly used by women with high semester ADS scores (defined as 75th percentile of scores). 1126 women (59.9%) used at least one medicine with anticholinergic properties. The median ADS score was 4 or 5 across all semesters. Most anticholinergic medicines used by women who had a high anticholinergic burden (ADS score, ≥ 9) had a low anticholinergic potency (ADS level 1). Increasing age, cardiovascular disease, and number of other medicines used were predictive of a higher anticholinergic burden. A high anticholinergic medicines burden in this group was driven by the use of multiple medicines with lower anticholinergic potency rather than the use of medicines with higher potency. This is a novel and important finding for clinical practice as doctors would readily identify the risk of a high anticholinergic burden for patients using high potency medicines, but may be less likely to identify this risk for users of multiple medicines with low anticholinergic potency.


Factors influencing the induction of high affinity antibodies to Plasmodium falciparum merozoite antigens and how affinity changes over time.
Understanding the functional characteristics of naturally acquired antibodies against P. falciparum merozoite antigens is crucial for determining the protective functions of antibodies. Affinity (measured as kd) of naturally acquired antibodies against two key targets of acquired immunity, EBA175 and PfRh2, was determined using Surface Plasmon Resonance (SPR) in a longitudinal survey in Nigeria. A majority of the participants, 79% and 67%, maintained stable antibody affinities to EBA175 and PfRh2, respectively, over time. In about 10% of the individuals, there was a reciprocal interaction with a reduction over time in antibody affinity for PfRh2 and an increase for EBA175. In general, PfRh2 elicited antibodies with higher affinity compared to EBA175. Individuals with higher exposure to malaria produced antibodies with higher affinity to both antigens. Younger individuals (5-15 years) produced comparable or higher affinity antibodies than adults (>15 years) against EBA175, but not for PfRh2. Correlation between total IgG (ELISA) and affinity varied between individuals, but PfRh2 elicited antibodies with a higher correlation in a majority of the participants. There was also a correlation between antibody inhibition of erythrocyte invasion by merozoites and PfRh2 affinity. This work gives new insights into the generation and maintenance of antibody affinity over time.


Analysing organisational context: case studies on the contribution of absorptive capacity theory to understanding inter-organisational variation in performance improvement.
Organisational context is frequently cited as an important consideration when implementing and evaluating quality improvement interventions in healthcare, but limited guidance is available on which aspects of context are most influential or modifiable. This paper examines how internal and external contextual factors mediate organisational-level performance improvement through applying the knowledge-based theory of absorptive capacity (AC). Three healthcare case studies are presented. Each case is a UK National Health Service organisation that had been identified as having performance problems. Qualitative data were collected through semi-structured interviews with general and clinical managers within the organisation and members of external teams supporting or overseeing performance improvement (n=22). Interview data were analysed using an existing AC framework from the literature. The organisation with the highest AC showed the quickest and most comprehensive performance improvement. Internal characteristics including strategic priorities, processes for managing information, communication and orientation to learning and development impacted on the organisation's ability to engage successfully with external stakeholders and make use of available knowledge. This enabled the organisation to thrive despite the challenging external environment. Lower levels of AC appeared to delay or limit the improvement trajectory. Developing a more detailed and nuanced understanding of how context influences improvement is an important step towards achieving more effective and sustainable quality improvement programmes in healthcare. AC, with its focus on knowledge and organisational learning, provides a useful way to explore the relationship between context and quality improvement and represents a potentially valuable area for future research and development.


Ventral hippocampal alpha 7 nicotinic receptor blockade and chronic nicotine effects on memory performance in the radial-arm maze.
Chronic nicotine administration has been shown to significantly improve working memory. Nicotinic involvement in memory function critically involves the ventral hippocampus. Local ventral hippocampal infusions of the nicotinic antagonists mecamylamine, dihydro-beta-erythroidine (DH beta E) and methyllycaconitine (MLA) significantly impair working memory. The impairment caused by hippocampal infusion of the alpha 4 beta 2 antagonist DH beta E is reversed by chronic systemic nicotine. This study determined the interaction of chronic systemic nicotine with acute ventral hippocampal infusions of the alpha 7 antagonist MLA. Adult female Sprague-Dawley rats were trained on an 8-arm radial maze working memory task. Then they underwent ventral hippocampal cannulation and received sc implants of minipumps delivering nicotine (0 or 5 mg/kg/day for 28 days). Acute ventral hippocampal infusions of MLA (0, 4.88, 14.64 and 43.92 microg/side) were given during 3-4 weeks of chronic nicotine. MLA caused a significant dose-related memory impairment. In the rats not receiving nicotine, the 14.64 and 43.92 microg/side MLA doses caused significant memory impairment. Chronic systemic nicotine exposure did not block the MLA-induced memory impairment. Comparing the current results with MLA with previous results with DH beta E, equimolar ventral hippocampal DH beta E more effectively impaired memory than MLA, but the DH beta E-induced impairment was more effectively reversed by chronic systemic nicotine administration.


First Report of Bacterial Leaf Spot of Basil Caused by Pseudomonas viridiflava in Hungary.
In April 2011, typical bacterial spot symptoms were observed on sweet basil plantlets (Ocimum basilicum L.) in a supermarket in Budapest, Hungary. Affected plants had dark brown-to-black lesions on the cotyledons. Spots on the leaves were first water soaked and then became necrotic and progressed inward from the margins. Symptoms were similar to those reported by Little et al. (3) on basil affected by Pseudomonas viridiflava. Bacteria consistently isolated from leaf lesions formed mucoid colonies with a green fluorescent pigment on King's B medium. Strains were gram negative. In LOPAT (levan-oxidase-potato rot-arginine dihydrolase-tobacco hypersensitivity) tests (2), all induced a hypersensitive reaction (HR) in tobacco (Nicotiana tabacum L. cv. White Burley) leaves (1), caused soft rot of potato tuber slices, and were negative for levan, oxidase, and arginine dihydrolase. Biochemical tests, API 20NE and API 50 CH (Biomérieux, Marcy l'Etoile, France), were also used for identification. The pathogenicity of three isolates was tested twice by injecting 20-day-old healthy basil plants with a bacterial suspension (107 CFU/ml). Controls were injected with sterile distilled water. Plants were kept at 25 to 28°C and 80 to 100% relative humidity. Forty-eight hours after inoculation, dark brown-to-black lesions were observed only on inoculated plants. The bacterium was reisolated from lesions of all plants tested, fulfilling Koch's postulates. No lesions were observed on controls. To identify the pathogen, a PCR technique was used. The 16SrDNA region was amplified with general bacterial primer pair (63f forward and 1389r reverse) (4) then the PCR products were cloned into Escherichia coli DH5α cells and a recombinant plasmid was sequenced by M13 forward and reverse primers. The sequence was deposited in GenBank (Accession No. HE585219). On the basis of the symptoms, biochemical tests, and 16SrDNA sequence homology (99% sequence similarity with a number of P. viridiflava isolates), the pathogen was identified as P. viridiflava. To our knowledge, this is the first report of bacterial leaf spot of basil in Hungary, which can seriously affect the basil production. References: (1) Z. Klement. Nature 199:299, 1963. (2) R. A. Lelliot et al. Appl. Bacteriol. 29:470, 1966. (3) E. L. Little et al. Plant Dis. 78:831, 1994. (4) A. M. Osborn et al. Environ. Microbiol. 2:39, 2000.


Guest-Dependent Stabilization of the Low-Spin State in Spin-Crossover Metal-Organic Frameworks.
Computer simulations are carried out to characterize the variation of spin-crossover (SCO) behavior of the prototypical {Fe(pz)[Pt(CN)4]} metal-organic framework (MOF) upon adsorption of chemically and structurally different guest molecules. A detailed analysis of both strength and anisotropy of guest molecule-framework interactions reveals direct correlations between the mobility of the guest molecules inside the MOF pores, the rotational mobility of the pyrazine rings of the framework, and the stabilization of the low-spin state of the material. On the basis of these correlations, precise molecular criteria are established for predicting the spin state of {Fe(pz)[Pt(CN)4]} upon guest adsorption. Finally, predictions of the SCO temperature upon adsorption of various toxic gases demonstrate that in silico modeling can provide fundamental insights and design principles for the development of spin-crossover MOFs for applications in gas detection and chemical sensing.


Tofacitinib Loaded Squalenyl Nanoparticles for Targeted Follicular Delivery in Inflammatory Skin Diseases.
Tofacitinib (TFB), a Janus kinase inhibitor, has shown excellent success off-label in treating various dermatological diseases, especially alopecia areata (AA). However, TFB's safe and targeted delivery into hair follicles (HFs) is highly desirable due to its systemic adverse effects. Nanoparticles (NPs) can enhance targeted follicular drug delivery and minimize interfollicular permeation and thereby reduce systemic drug exposure. In this study, we report a facile method to assemble the stable and uniform 240 nm TFB loaded squalenyl derivative (SqD) nanoparticles (TFB SqD NPs) in aqueous solution, which allowed an excellent loading capacity (LC) of 20%. The SqD NPs showed an enhanced TFB delivery into HFs compared to the aqueous formulations of plain drug in an ex vivo pig ear model. Furthermore, the therapeutic efficacy of the TFB SqD NPs was studied in a mouse model of allergic dermatitis by ear swelling reduction and compared to TFB dissolved in a non-aqueous mixture of acetone and DMSO (7:1 v/v). Whereas such formulation would not be acceptable for use in the clinic, the TFB SqD NPs dispersed in water illustrated a better reduction in inflammatory effects than plain TFB's aqueous formulation, implying both encouraging good in vivo efficacy and safety. These findings support the potential of TFB SqD NPs for developing a long-term topical therapy of AA.


Honorary Authorships in Surgical Literature.
Honorary authorship (HA) is defined as an enlisted co-author who did not make sufficient contributions to merit being included as a co-author according to the ICMJE guidelines on authorship. It is unknown if HA is present in the surgical literature. We analysed studies published in 2016 in five journals with the highest impact factor in general surgery. All original articles, reviews and clinical trials with more than one author were included. Corresponding authors of these manuscripts received an online survey by email. The survey consisted of three parts focussing on demographics, knowledge and application of the ICMJE guidelines, and deciding authorship. In total, 320 of the 1037 surveys were completed (30.9%). Two hundred and seventy-two (88.6%) of the corresponding authors were aware of the ICMJE authorship guidelines, and 203 (66.3%) were aware of the general issue of honorary authorship. One hundred and thirty-five (44.0%) responders reported at least one co-author who only performed tasks which should not merit actual authorship according to the ICMJE guidelines. Furthermore, only 46 (15.0%) of the responders believed that a co-author listed for their article did not make sufficient contribution to merit being included as co-author. No significant differences were found between the journals investigated. Despite ICMJE guidelines to reduce HA, the prevalence may still exist to a higher level than preferable. The authors plead for more transparent authorship systems in which journal editors and senior department members take more responsibility into enforcement of the ICMJE guidelines.


Integrins αvβ3 and α4β1 act as coreceptors for fractalkine, and the integrin-binding defective mutant of fractalkine is an antagonist of CX3CR1.
The membrane-bound chemokine fractalkine (FKN, CX3CL1) on endothelial cells plays a role in leukocyte trafficking. The chemokine domain (FKN-CD) is sufficient for inducing FKN signaling (e.g., integrin activation), and FKN-CD binds to its receptor CX3CR1 on leukocytes. Whereas previous studies suggest that FKN-CD does not directly bind to integrins, our docking simulation studies predicted that FKN-CD directly interacts with integrin α(v)β(3). Consistent with this prediction, we demonstrated that FKN-CD directly bound to α(v)β(3) and α(4)β(1) at a very high affinity (K(D) of 3.0 × 10(-10) M to α(v)β(3) in 1 mM Mn(2+)). Also, membrane-bound FKN bound to integrins α(v)β(3) and α(4)β(1), suggesting that the FKN-CD/integrin interaction is biologically relevant. The binding site for FKN-CD in α(v)β(3) was similar to those for other known α(v)β(3) ligands. Wild-type FKN-CD induced coprecipitation of integrins and CX3CR1 in U937 cells, suggesting that FKN-CD induces ternary complex formation (CX3CR1, FKN-CD, and integrin). Based on the docking model, we generated an integrin-binding defective FKN-CD mutant (the K36E/R37E mutant). K36E/R37E was defective in ternary complex formation and integrin activation, whereas K36E/R37E still bound to CX3CR1. These results suggest that FKN-CD binding to CX3CR1 is not sufficient for FKN signaling, and that FKN-CD binding to integrins as coreceptors and the resulting ternary complex formation are required for FKN signaling. Notably, excess K36E/R37E suppressed integrin activation induced by wild-type FKN-CD and effectively suppressed leukocyte infiltration in thioglycollate-induced peritonitis. These findings suggest that K36E/R37E acts as a dominant-negative CX3CR1 antagonist and that FKN-CD/integrin interaction is a novel therapeutic target in inflammatory diseases.


Evaluating User Engagement with a Reminiscence App Using Cross-Comparative Analysis of User Event Logs and Qualitative Data.
The aim of this study was to evaluate the usage of a reminiscence app by people living with dementia and their family carers, by comparing event log data generated from app usage alongside the qualitative experience of the process. A cross-comparative analysis of electronic event logging data with qualitative interview data was conducted. Electronic event logging data were obtained for 28 participating dyads (n = 56) and the interview sample comprised 14 people living with dementia and 16 family carers (n = 30). A thematic analysis framework was used in the analysis of interview transcripts and the identification of recurrent themes. The cross-comparison of electronic event log data and qualitative data revealed 25 out of 28 dyads regularly engaged with a reminiscence app, with the analysis of usage patterns revealing four clusters classifying different levels of user engagement. The cross-comparison of data revealed that the nature of the relationship was a significant factor in ongoing user engagement. The comparative analysis of the electronic event logs as "ground truth" in combination with the qualitative lived experience can provide a deeper understanding on the usage of a reminiscence app for those living with dementia and their family carers. This work not only shows the benefits of using automated event log data mining but also shows its clear limitations without using complementary qualitative data analysis. As such, this work also provides key insights into using mixed methods for evaluating human-computer interaction technologies.


Speckle-free, shaded 3D images produced by computer-generated holography.
A hologram display technique that provides speckle-free, shaded reconstructed images is proposed. A three-dimensional object consists of object points; these object points are divided into plural object point groups that are generated in a time-sequential manner. Each object point group consists of a two-dimensional (2D) array of object points that are separated so as to prevent interference among them. Each object point group is generated by displaying a 2D array of zone plates on a high-speed spatial light modulator (SLM). The amplitude distribution of the zone plates is modulated two-dimensionally based on Phong shading to shade the reconstructed images. The 2D amplitude distribution of the zone plates is decomposed into multiple binary patterns that are displayed by the SLM in a time-sequential manner. The proposed method is experimentally verified.


Perceptions of cisplatin-related toxicity among ovarian cancer patients and gynecologic oncologists.
We conducted a pilot study to evaluate issues related to chemotherapy-induced toxicities by eliciting assessments of toxicity from women with advanced stage ovarian cancer and gynecologic oncologists. Fifteen ovarian cancer patients and ten gynecologic oncologists completed the survey exercises. All patients surveyed had received at least six courses of a cisplatin-containing chemotherapy regimen. For both patients and physicians, there was good face validity to the utility exercise as assessments of health states with cisplatin were (1) consistently associated with less favorable assessments than the health state with no toxicity and (2) neurotoxicity was viewed less favorably than either ototoxicity or nephrotoxicity. While the 15 patients as a group viewed health states with toxicity more favorably than physicians (P < 0.05 for each toxicity), patient assessments varied, depending on individual experiences with cisplatin. Physician assessments of toxicity were most similar to those obtained from patients who had not experienced cisplatin toxicity and were less favorable than those elicited from patients who had experienced any toxicity. In deciding upon therapeutic strategies, women with advanced stage ovarian cancer and treating physicians markedly differ in their assessment of the impact of specific toxicities on quality of life.


Identification of macrophage and stress-induced proteins of Mycobacterium tuberculosis.
Using phosphorimager technology to quantitate differences in protein expression, we have investigated the modulation of protein synthesis by Mycobacterium tuberculosis in response to intracellular residence in human macrophages and, for comparison, in response to various stress conditions during extracellular growth. Proteins of M. tuberculosis growing intracellularly in human THP-1 cells and extracellularly in broth were labeled with [35S]methionine; during intracellular growth, host cell protein synthesis was inhibited with cycloheximide. The metabolically labeled proteins were separated by two-dimensional gel electrophoresis and quantitatively analyzed. Intracellular residence in macrophages induced a profound change in the overall phenotype of M. tuberculosis. The expression of at least 16 M. tuberculosis proteins was induced (at least a twofold increase compared with growth in broth) and 28 proteins repressed (at least a twofold decrease). Many of the phenotypic changes in protein expression induced during intracellular growth occurred during extracellular growth in response to stress conditions including heat-shock, low pH, and H2O2. However, the pattern of induced and repressed proteins was unique to each stress condition. Of the 16 macrophage-induced proteins, 6 were absent during extracellular growth under both normal and stress conditions. Such proteins are potential virulence determinants and/or they may be important in the cell-mediated and protective immune response to M. tuberculosis infection.


Antifungal effects of phytocompounds on Candida species alone and in combination with fluconazole.
Invasive fungal infections caused by Candida spp. remain the most predominant nosocomial fungal infections. Owing to the increased use of antifungal agents, resistance of Candida spp. to antimycotics has emerged frequently, especially to fluconazole (FLC). To cope with this issue, new efforts have been dedicated to discovering novel antimycotics or new agents that can enhance the susceptibility of Candida spp. to existing antimycotics. The secondary metabolites of plants represent a large library of compounds that are important sources for new drugs or compounds suitable for further modification. Research on the anti-Candida activities of phytocompounds has been carried out in recent years and the results showed that a series of phytocompounds have anti-Candida properties, such as phenylpropanoids, flavonoids, terpenoids and alkaloids. Among these phytocompounds, some displayed potent antifungal activity, with minimum inhibitory concentrations (MICs) of ≤8 µg/mL, and several compounds were even more effective against drug-resistant Candida spp. than FLC or itraconazole (e.g. honokiol, magnolol and shikonin). Interestingly, quite a few phytocompounds not only displayed anti-Candida activity alone but also synergised with FLC against Candida spp., even leading to a reversal of FLC resistance. This review focuses on summarising the anti-Candida activities of phytocompounds as well as the interactions of phytocompounds with FLC. In addition, we briefly overview the synergistic mechanisms and present the structure of the antimycotic phytocompounds. Hopefully, this analysis will provide insight into antifungal agent discovery and new approaches against antifungal drug resistance.


Clinical study on the reasons for and location of failures of metal-ceramic restorations and survival of repairs.
A recently introduced technique, the Cojet system, using SiOx (Al2O3 coated with silisic acid), provides ultrafine mechanical retention by sandblasting, as well as a chemicophysical bond between the metal-ceramic and the composite resin. This study determined the reasons for and locations of failures of metal-ceramic restorations and evaluated the survival of the intraoral silica-coating system used for the repair of failed restorations. A total of 153 patients possessing 289 fractured crowns were involved in this study; 255 of these fractures were on fixed partial dentures, whereas 34 were on single crowns. The mean observation period was 34.6 months. The materials used for the repair process were ESPE-Sil for silane; Visiogem, Sinfony, and Dentacolor as opaquers; and Pertac II Aplitip, Sinfony, and Charisma as repairing composite resins. The majority of the failures (65%) occurred in the anterior region. Sixty percent of the failures were observed at the labial, 27% at the buccal, 5% at the incisal, and 8% at the occlusal regions. The fractures were mainly in the maxilla (75%), predominantly at the labial surface. The overall cumulative survival rates of the repairs (89%) showed that the first failures happened mostly from 1 week to 3 months after the repair, without any difference between the composite resins. The original failures happened mainly during chewing function or because of accidents, iatrogenic factors, and surgical operations, whereas secondary failures after repair using intraoral silica coating and composite resin were due to trauma, chewing function, or lack of rubber dam application.


A curcumin-loaded liquid crystal precursor mucoadhesive system for the treatment of vaginal candidiasis.
Women often develop vaginal infections that are caused primarily by organisms of the genus Candida. The current treatments of vaginal candidiasis usually involve azole-based antifungals, though fungal resistance to these compounds has become prevalent. Therefore, much attention has been given to molecules with antifungal properties from natural sources, such as curcumin (CUR). However, CUR has poor solubility in aqueous solvents and poor oral bioavailability. This study attempted to overcome this problem by developing, characterizing, and evaluating the in vitro antifungal action of a CUR-loaded liquid crystal precursor mucoadhesive system (LCPM) for vaginal administration. A low-viscosity LCPM (F) consisting of 40% wt/wt polyoxpropylene-(5)-polyoxyethylene-(20)-cetyl alcohol, 50% wt/wt oleic acid, and 10% wt/wt chitosan dispersion at 0.5% with the addition of 16% poloxamer 407 was developed to take advantage of the lyotropic phase behavior of this formulation. Notably, F could transform into liquid crystal systems when diluted with artificial vaginal mucus at ratios of 1:3 and 1:1 (wt/wt), resulting in the formation of F30 and F100, respectively. Polarized light microscopy and rheological studies revealed that F behaved like an isotropic formulation, whereas F30 and F100 behaved like an anisotropic liquid crystalline system (LCS). Moreover, F30 and F100 presented higher mucoadhesion to porcine vaginal mucosa than F. The analysis of the in vitro activity against Candida albicans revealed that CUR-loaded F was more potent against standard and clinical strains compared with a CUR solution. Therefore, the vaginal administration of CUR-loaded LCPMs represents a promising platform for the treatment of vaginal candidiasis.


Site-specific labeling of T7 DNA polymerase with a conformationally sensitive fluorophore and its use in detecting single-nucleotide polymorphisms.
Like most enzymes, DNA polymerases undergo a large conformational change on the binding of a correct nucleotide. To determine how the conformational change contributes to substrate specificity, we labeled the T7 DNA polymerase with a conformationally sensitive fluorophore at a position that provides a signal coincident with structural changes following nucleotide binding and distinguishes correct base pairs from incorrect ones by the sign of the fluorescence change. Here we describe methods to document that only one site on the polymerase was labeled with the fluorophore based on mass spectral analysis of tryptic peptides. In addition, we show by equilibrium titrations of opposing signals that mismatches and correct bases compete for the same site. This analysis forms an essential basis for characterization of a fluorescently labeled enzyme intended for mechanistic studies. Finally, we show that the labeled enzyme can be used to identify single-nucleotide mutations in a procedure that could be automated.


Invalidation of TASK1 potassium channels disrupts adrenal gland zonation and mineralocorticoid homeostasis.
TASK1 (KCNK3) and TASK3 (KCNK9) are two-pore domain potassium channels highly expressed in adrenal glands. TASK1/TASK3 heterodimers are believed to contribute to the background conductance whose inhibition by angiotensin II stimulates aldosterone secretion. We used task1-/- mice to analyze the role of this channel in adrenal gland function. Task1-/- exhibited severe hyperaldosteronism independent of salt intake, hypokalemia, and arterial 'low-renin' hypertension. The hyperaldosteronism was fully remediable by glucocorticoids. The aldosterone phenotype was caused by an adrenocortical zonation defect. Aldosterone synthase was absent in the outer cortex normally corresponding to the zona glomerulosa, but abundant in the reticulo-fasciculata zone. The impaired mineralocorticoid homeostasis and zonation were independent of the sex in young mice, but were restricted to females in adults. Patch-clamp experiments on adrenal cells suggest that task3 and other K+ channels compensate for the task1 absence. Adrenal zonation appears as a dynamic process that even can take place in adulthood. The striking changes in the adrenocortical architecture in task1-/- mice are the first demonstration of the causative role of a potassium channel in development/differentiation.


A voxel-wise assessment of growth differences in infants developing autism spectrum disorder.
Autism Spectrum Disorder (ASD) is a phenotypically and etiologically heterogeneous developmental disorder typically diagnosed around 4 years of age. The development of biomarkers to help in earlier, presymptomatic diagnosis could facilitate earlier identification and therefore earlier intervention and may lead to better outcomes, as well as providing information to help better understand the underlying mechanisms of ASD. In this study, magnetic resonance imaging (MRI) scans of infants at high familial risk, from the Infant Brain Imaging Study (IBIS), at 6, 12 and 24 months of age were included in a morphological analysis, fitting a mixed-effects model to Tensor Based Morphometry (TBM) results to obtain voxel-wise growth trajectories. Subjects were grouped by familial risk and clinical diagnosis at 2 years of age. Several regions, including the posterior cingulate gyrus, the cingulum, the fusiform gyrus, and the precentral gyrus, showed a significant effect for the interaction of group and age associated with ASD, either as an increased or a decreased growth rate of the cerebrum. In general, our results showed increased growth rate within white matter with decreased growth rate found mostly in grey matter. Overall, the regions showing increased growth rate were larger and more numerous than those with decreased growth rate. These results detail, at the voxel level, differences in brain growth trajectories in ASD during the first years of life, previously reported in terms of overall brain volume and surface area.


Potential impact of task-shifting on costs of antiretroviral therapy and physician supply in Uganda.
Lower-income countries face severe health worker shortages. Recent evidence suggests that this problem can be mitigated by task-shifting--delegation of aspects of health care to less specialized health workers. We estimated the potential impact of task-shifting on costs of antiretroviral therapy (ART) and physician supply in Uganda. The study was performed at the Infectious Diseases Institute (IDI) clinic, a large urban HIV clinic. We built an aggregate cost-minimization model from societal and Ministry of Health (MOH) perspectives. We compared physician-intensive follow-up (PF), the standard of care, with two methods of task-shifting: nurse-intensive follow-up (NF) and pharmacy-worker intensive follow-up (PWF). We estimated personnel and patient time use using a time-motion survey. We obtained unit costs from IDI and the literature. We estimated physician personnel impact by calculating full time equivalent (FTE) physicians saved. We made national projections for Uganda. Annual mean costs of follow-up per patient were $59.88 (societal) and $31.68 (medical) for PF, $44.58 (societal) and $24.58 (medical) for NF and $18.66 (societal) and $10.5 (medical) for PWF. Annual national societal ART follow-up expenditure was $5.92 million using PF, $4.41 million using NF and $1.85 million using PWF, potentially saving $1.51 million annually by using NF and $4.07 million annually by using PWF instead of PF. Annual national MOH expenditure was $3.14 million for PF, $2.43 million for NF and $1.04 for PWF, potentially saving $0.70 million by using NF and $2.10 million by using PWF instead of PF. Projected national physician personnel needs were 108 FTE doctors to implement PF and 18 FTE doctors to implement NF or PWF. Task-shifting from PF to NF or PWF would potentially save 90 FTE physicians, 4.1% of the national physician workforce or 0.3 FTE physicians per 100,000 population. Task-shifting results in substantial cost and physician personnel savings in ART follow-up in Uganda and can contribute to mitigating the heath worker crisis.