PCR targeting Plasmodium mitochondrial genome of DNA extracted from dried blood on filter paper compared to whole blood.
Monitoring mortality and morbidity attributable to malaria is paramount to achieve elimination of malaria. Diagnosis of malaria is challenging and PCR is a reliable method for identifying malaria with high sensitivity. However, blood specimen collection and transport can be challenging and obtaining dried blood spots (DBS) on filter paper by finger-prick may have advantages over collecting whole blood by venepuncture. DBS and whole blood were collected from febrile children admitted at the general paediatric wards at a referral hospital in Dar es Salaam, Tanzania. DNA extracted from whole blood and from DBS was tested with a genus-specific PCR targeting the mitochondrial Plasmodium genome. Positive samples by PCR of DNA from whole blood were tested with species-specific PCR targeting the 18S rRNA locus, or sequencing if species-specific PCR was negative. Rapid diagnostic test (RDT) and thin blood smear microscopy was carried out on all patients where remnant whole blood and a blood slide, respectively, were available. Positivity of PCR was 24.5 (78/319) and 11.2% (52/442) by whole blood and DBS, respectively. All samples positive on DBS were also positive on Plasmodium falciparum species-specific PCR. All RDT positive cases were also positive by DBS PCR. All but three cases with positive blood slides were also positive by DBS. In this study, PCR for malaria mitochondrial DNA extracted from whole blood was more sensitive than from DBS. However, DBS are a practical alternative to whole blood and detected approximately the same number of cases as RDTs and, therefore, remain relevant for research purposes.


Urinary citrate, bone resorption and intestinal alkali absorption in stone formers with fasting hypercalciuria.
Reduced citrate in urine and increased fasting excretion of calcium are abnormalities frequently reported in stone forming (SF) patients. Increased dietary acid (or reduced alkali) introduction or absorption may be a potential cause of both these pathological findings. To test this hypothesis, we studied 64 SF patients (32 with fasting hypercalciuria (FH) and 32 without FH (NFH)). After a basal evaluation for nephrolithiasis, while on a 500 mg calcium diet, they were evaluated for: (1) daily intestinal alkali absorption (IAA), by urinary electrolyte excretion; (2) basal concentrations of PTH, calcitonin (CT) and 1,25(OH)2-VitD; (3) oral calcium load for evaluation of changes in calcium and hydroxyproline urinary excretions; (4) intestinal calcium absorption (18 patients), with double curve analysis (stable Sr as tracer); and (5) changes in citrate excretion after an alkali load (50 mEq of a mixture of calcium gluconate, lactate and carbonate) in 10 patients. The results demonstrated: (1) FH stone formers had reduced citrate excretion and lower mean IAA levels than NFH stone formers; (2) FH stone formers also had higher bone resorption levels with lower PTH and higher CT levels; (3) IAA levels were related to both citrate excretion and bone turnover indices; and (4) the increases in citrate excretion after oral alkali load were strictly related to basal IAA values (index of alkali absorption and/or generation after oral load), demonstrating that a different absorptive capacity of alkali rather than a different dietary content may underlie these metabolic abnormalities.


Predictors of Participation in a Fire Department Community Canvassing Program.
An urban fire department has been distributing free smoke alarms for more than 30 years. A community-academic partnership was developed to conduct a community intervention trial as part of the fire department's home visiting program. The trial comprised 170 canvassing events held across 12 census tracts; half of the census tracts were assigned to the treatment condition and received prepromotion of the home visit events. The objectives of this analysis were to identify environmental and programmatic predictors of 1) whether someone would be at home at the time of a visit, and 2) if at home, whether the resident would participate. A separate multilevel analysis was conducted to address each objective. The canvassing event served as the first level to account for variation in implementation of the program, with the census tract as the second level. All environmental and program characteristics were included as fixed effects in both models. Throughout 170 events, 8080 eligible residential addresses were visited, of which 3216 had someone at home, and 2197 homes participated in the program. Canvassing events held on weekends and during the evening hours was associated with higher odds of a resident being at home. Canvassing events without rain and held in the treatment census tract areas was associated with higher odds of resident participation. Environmental and programmatic factors can impact the reach of home visiting programs. These findings can contribute to emerging best practices for fire department home visiting programs.


Genetic epidemiologic studies on age-specified traits. NIA Aging and Genetic Epidemiology Working Group.
This commentary calls attention to the value of combining genetic and epidemiologic methods in studies to understand the determinants of two crucial aspects of aging: ages at which certain outcomes (e.g., disease, mortality) occur and rates of change with age of individual's characteristics (e.g., physiologic functions, disease risk factors). Inclusion of age in the specification of traits in genetic epidemiologic studies could lead to improved strategies to increase healthy life expectancy and evaluate individuals' risk for age-related morbidity. Special issues that make genetic epidemiologic approaches important for studies of age-specified phenomena as well as opportunities and challenges for such studies are discussed, including study designs, sampling frames, databases, analytic tools, and related methodological issues. This commentary is based on a report prepared by the Aging and Genetic Epidemiology Working Group, convened by the National Institute on Aging to review opportunities for research on the genetic epidemiology of aging-related outcomes. The report, which contains more extensive discussion, literature review, and references, is available on the World Wide Web at http://www.nih.gov/nia/conferences/GeneticReport111199.htm.


Nodal root diameter and node number in maize (Zea mays L.) interact to influence plant growth under nitrogen stress.
Under nitrogen limitation, plants increase resource allocation to root growth relative to shoot growth. The utility of various root architectural and anatomical phenotypes for nitrogen acquisition are not well understood. Nodal root number and root cross-sectional area were evaluated in maize in field and greenhouse environments. Nodal root number and root cross-sectional area were inversely correlated under both high and low nitrogen conditions. Attenuated emergence of root nodes, as opposed to differences in the number of axial roots per node, was associated with substantially reduced root number. Greater root cross-sectional area was associated with a greater stele area and number of cortical cell files. Genotypes that produced few, thick nodal roots rather than many, thin nodal roots had deeper rooting and better shoot growth in low nitrogen environments. Fewer nodal roots offset the respiratory and nitrogen costs of thicker diameter roots, since total nodal root respiration and nitrogen content was similar for genotypes with many, thin and few, thick nodal roots. We propose that few, thick nodal roots may enable greater capture of deep soil nitrogen and improve plant performance under nitrogen stress. Synergistic interactions between an architectural and anatomical trait may be an important strategy for nitrogen acquisition. Understanding trait interactions among different root nodes has important implications in for improving crop nutrient uptake and stress tolerance.


The easy road to genome-wide medium density SNP screening in a non-model species: development and application of a 10 K SNP-chip for the house sparrow (Passer domesticus).
With the advent of next generation sequencing, new avenues have opened to study genomics in wild populations of non-model species. Here, we describe a successful approach to a genome-wide medium density Single Nucleotide Polymorphism (SNP) panel in a non-model species, the house sparrow (Passer domesticus), through the development of a 10 K Illumina iSelect HD BeadChip. Genomic DNA and cDNA derived from six individuals were sequenced on a 454 GS FLX system and generated a total of 1.2 million sequences, in which SNPs were detected. As no reference genome exists for the house sparrow, we used the zebra finch (Taeniopygia guttata) reference genome to determine the most likely position of each SNP. The 10 000 SNPs on the SNP-chip were selected to be distributed evenly across 31 chromosomes, giving on average one SNP per 100 000 bp. The SNP-chip was screened across 1968 individual house sparrows from four island populations. Of the original 10 000 SNPs, 7413 were found to be variable, and 99% of these SNPs were successfully called in at least 93% of all individuals. We used the SNP-chip to demonstrate the ability of such genome-wide marker data to detect population sub-division, and compared these results to similar analyses using microsatellites. The SNP-chip will be used to map Quantitative Trait Loci (QTL) for fitness-related phenotypic traits in natural populations.


Reflectance measurements and optical constants in the extreme ultraviolet for thin films of ion-beam-deposited SiC, Mo, Mg2Si, and InSb and of evaporated Cr.
Reflectance measurements and optical constants of thin films of ion-beam-deposited SiC, Mo, Mg(2)Si, and InSb and of evaporated Cr have been measured in the extreme-ultraviolet (EUV) spectral region from 49.0 to 200.0 nm. In this spectral region no optical constant data were available for materials deposited by ion-beam deposition. We compared our data with those for bulk samples and for thin films prepared by different techniques. The goal of this research has been to study candidate materials for multilayer coatings in the EUV.


Sucralfate reduces the gastrointestinal absorption of norfloxacin.
The effect of sucralfate on the bioavailability of norfloxacin after single 400-mg doses of norfloxacin was evaluated in eight healthy males. Subjects received each of the following treatments in random sequence: (i), norfloxacin, 400 mg alone; (ii) sucralfate, 1 g, concurrently with norfloxacin, 400 mg; and (iii) sucralfate, 1 g, followed by norfloxacin, 400 mg, 2 h later. One day before administration of treatments 2 and 3, 1 g of sucralfate was given at 7 a.m., 11 a.m., 5 p.m., and 10 p.m. Blood samples were collected immediately before the norfloxacin dose and at 0.25, 0.5, 0.75, 1.0, 1.5, 2, 3, 4, 6, 8, 12, and 24 h postdose. Urine was collected in divided intervals: from 0 to 12, from 12 to 24, and from 24 to 48 h. Norfloxacin concentrations in plasma and urine were determined by high-performance liquid chromatography. Mean area under the plasma concentration-versus-time curve extrapolated to infinity decreased significantly (P less than 0.001) after norfloxacin was given with and 2 h after sucralfate. The relative bioavailabilities were 1.8% when norfloxacin was taken with sucralfate and 56.6% when it was taken 2 h after sucralfate. After norfloxacin was given alone, the mean norfloxacin concentrations in urine collected during intervals of 0 to 12, 12 to 24, and 24 to 28 h were 118.9 +/- 72.3, 18.8 +/- 12.5, and 2.4 +/- 2.2 micrograms/ml, respectively. After norfloxacin was given with sucralfate, however, the mean norfloxacin concentrations in urine collected during the same time intervals were 6.8 +/- 4.7, 1.8 +/- 1.4, and 0 +/- 0 microgram/ml, respectively. Because of low pH and relatively high magnesium concentration in urine, susceptibilities of bacteria in urine are 8- to 32-fold lower than in broth. This fact, in combination with the reduced bioavailability of norfloxacin in the presence of sucralfate or antacids, is likely to result in treatment failure. The effect of sucralfate given after norfloxacin was not examined, nor was the effect of sucralfate given more than 2 h before norfloxacin. Administration or norfloxacin with sucralfate should therefore by avoided.


Meta-analysis of randomized controlled trials on cognitive effects of Bacopa monnieri extract.
Bacopa monnieri has a long history in Ayurvedic medicine for neurological and behavioral defects. To assess its efficacy in improving cognitive function. MEDLINE, EMBASE, CINAHL, AMED, Cochrane Central of clinical trial, WHO registry, Thai Medical Index, Index Medicus Siriraj library and www.clinicaltrial.gov were searched from the inception date of each database to June 2013 using scientific and common synonyms of Bacopa monnieri, cognitive performance or memory. The reference lists of retrieved articles were also reviewed. Randomized, placebo controlled human intervention trials on chronic ≥ 12 weeks dosing of standardized extracts of Bacopa monnieri without any co-medication were included in this study. The methodological quality of studies was assessed using Cochrane's risk of bias assessment and Jadad's quality scales. The weighted mean difference and 95% confidence interval (95% CI) were performed using the random-effects model of the Dersimonian-Laird method. Nine studies met the inclusion criteria using 518 subjects. Overall quality of all included trials was low risk of bias and quality of reported information was high. Meta-analysis of 437 eligible subjects showed improved cognition by shortened Trail B test (-17.9 ms; 95% CI -24.6 to -11.2; p<0.001) and decreased choice reaction time (10.6 ms; 95% CI -12.1 to -9.2; p<0.001). This meta-analysis suggests that Bacopa monnieri has the potential to improve cognition, particularly speed of attention but only a large well designed 'head-to-head' trial against an existing medication will provide definitive data on its efficacy on healthy or dementia patients using a standardized preparation.


Current evidence and opportunities in child and adolescent public mental health: a research review.
A public mental health lens is increasingly required to better understand the complex and multifactorial influences of interpersonal, community and institutional systems on the mental health of children and adolescents. This research review (1) provides an overview of public mental health and proposes a new interactional schema that can guide research and practice, (2) summarises recent evidence on public mental health interventions for children and adolescents, (3) highlights current challenges for this population that might benefit from additional attention and (4) discusses methodological and conceptual hurdles and proposes potential solutions. In our evidence review, a broad range of universal, selective and indicated interventions with a variety of targets, mechanisms and settings were identified, some of which (most notably parenting programmes and various school-based interventions) have demonstrated small-to-modest positive effects. Few, however, have achieved sustained mental health improvements. There is an opportunity to re-think how public mental health interventions are designed, evaluated and implemented. Deliberate design, encompassing careful consideration of the aims and population-level impacts of interventions, complemented by measurement that embraces complexity through more in-depth characterisation, or 'phenotyping', of interpersonal and environmental elements is needed. Opportunities to improve child and adolescent mental health outcomes are gaining unprecedented momentum. Innovative new methodology, heightened public awareness, institutional interest and supportive funding can enable enhanced study of public mental health that does not shy away from complexity.


The 'Pants-Sign': A Predictor for Falling in People with Parkinson's Disease?
A history of falls is the most established predictor of future falls in people with Parkinson's disease (PD). However, predicting a first fall remains challenging. To assess whether experiencing difficulties putting on pants while standing is a viable predictor of future falling, and specifically a first fall, in persons with PD. We define this 'Pants-sign' as people who resort to putting on their pants only while seated. 264 persons with PD were included. Information on the Pants-sign, history of falls, disease severity (MDS-UPDRS part III), freezing of gait (N-FOGQ > 0), cognitive function (MoCA), self-reported disability (Schwab & England scale), health-related quality of life (SF-12), Timed-Up-and-Go, and one-legged stance were determined at baseline and after one-year follow-up. The association between the Pants-sign and future falling was examined by univariate logistic regression analysis. A multivariate step-wise logistic regression with forward selection was employed to identify the strongest associations in the entire cohort and a sub-cohort of people without falls in the year prior to baseline. The Pants-sign was univariably associated with a future fall (OR = 2.406, 95% CI [1.313-4.409], p = 0.004]), but was not an independent predictor in the multivariate logistic regression; predictors were higher MDS-UPDRS part III scores (OR = 1.088, 95% CI [1.056-1.121], p < 0.001] and history of falls (OR = 5.696, 95% CI [2.650-12.243], p≤0.001]. For the sub-cohort of people without falls in the previous year (n = 189), the Pants-sign was not associated with future falls. The Pants-sign is simple to assess and is associated with future falling in PD but is not an independent predictor.


American Cancer Society guidelines on nutrition and physical activity for cancer prevention: Reducing the risk of cancer with healthy food choices and physical activity.
The American Cancer Society (ACS) has set aggressive challenge goals for the nation to decrease cancer incidence and mortality--and to improve the quality of life of cancer survivors--by the year 2015. To address these critical goals, the ACS publishes the Nutrition and Physical Activity Guidelines to serve as a foundation for its communication, policy, and community strategies and ultimately, to affect dietary and physical activity patterns among Americans. These guidelines, published every five years, are developed by a national panel of experts in cancer research, prevention, epidemiology, public health, and policy, and as such, they represent the most current scientific evidence related to dietary and activity patterns and cancer risk. The American Cancer Society guidelines include recommendations for individual choices regarding diet and physical activity patterns, but those choices occur within a community context that either facilitates or interferes with healthy behaviors. Therefore, this committee presents one key recommendation for community action to accompany the four recommendations for individual choices for nutrition and physical activity to reduce cancer risk. This recommendation for community action underscores just how important community measures are to the support of healthy behaviors by means of increasing access to healthful food choices and opportunities to be physically active. The ACS guidelines are consistent with guidelines from the American Heart Association for the prevention of coronary heart disease as well as for general health promotion, as defined by the Department of Health and Human Services' 2000 Dietary Guidelines for Americans.


Discovery of Metabolic Biomarkers for Duchenne Muscular Dystrophy within a Natural History Study.
Serum metabolite profiling in Duchenne muscular dystrophy (DMD) may enable discovery of valuable molecular markers for disease progression and treatment response. Serum samples from 51 DMD patients from a natural history study and 22 age-matched healthy volunteers were profiled using liquid chromatography coupled to mass spectrometry (LC-MS) for discovery of novel circulating serum metabolites associated with DMD. Fourteen metabolites were found significantly altered (1% false discovery rate) in their levels between DMD patients and healthy controls while adjusting for age and study site and allowing for an interaction between disease status and age. Increased metabolites included arginine, creatine and unknown compounds at m/z of 357 and 312 while decreased metabolites included creatinine, androgen derivatives and other unknown yet to be identified compounds. Furthermore, the creatine to creatinine ratio is significantly associated with disease progression in DMD patients. This ratio sharply increased with age in DMD patients while it decreased with age in healthy controls. Overall, this study yielded promising metabolic signatures that could prove useful to monitor DMD disease progression and response to therapies in the future.


Genomes and Transcriptomes of Duckweeds.
Duckweeds (Lemnaceae family) are the smallest flowering plants that adapt to the aquatic environment. They are regarded as the promising sustainable feedstock with the characteristics of high starch storage, fast propagation, and global distribution. The duckweed genome size varies 13-fold ranging from 150 Mb in Spirodela polyrhiza to 1,881 Mb in Wolffia arrhiza. With the development of sequencing technology and bioinformatics, five duckweed genomes from Spirodela and Lemna genera are sequenced and assembled. The genome annotations discover that they share similar protein orthologs, whereas the repeat contents could mainly explain the genome size difference. The gene families responsible for cell growth and expansion, lignin biosynthesis, and flowering are greatly contracted. However, the gene family of glutamate synthase has experienced expansion, indicating their significance in ammonia assimilation and nitrogen transport. The transcriptome is comprehensively sequenced for the genera of Spirodela, Landoltia, and Lemna, including various treatments such as abscisic acid, radiation, heavy metal, and starvation. The analysis of the underlying molecular mechanism and the regulatory network would accelerate their applications in the fields of bioenergy and phytoremediation. The comparative genomics has shown that duckweed genomes contain relatively low gene numbers and more contracted gene families, which may be in parallel with their highly reduced morphology with a simple leaf and primary roots. Still, we are waiting for the advancement of the long read sequencing technology to resolve the complex genomes and transcriptomes for unsequenced Wolffiella and Wolffia due to the large genome sizes and the similarity in their polyploidy.


The thermo- and acido-stable ORF-99 from the archaeal virus AFV1.
Acidianus Filamentous Virus 1 (AFV1), isolated from acidic hot springs, is an enveloped lipid-containing archaeal filamentous virus with a linear double-stranded DNA genome. It infects Acidianus, which is a hyperthermostable archaea growing at 85 degrees C and acidic pHs, below pH 3. AFV1-99, a protein of 99 amino acids of unknown function, has homologues in the archaeal virus families Lipothrixviridae and Rudiviridae. We determined the crystal structure of AFV1-99 at 2.05 A resolution. AFV1-99 has a new fold, is hyperthermostable (up to 95 degrees C) and resists to extreme pH (between pH 0 and 11) and to the combination of high temperature (95 degrees C) and low pH (pH 0). It possesses characteristics of hyperthermostable proteins, such as a high content of charged residues.


Should we listen to our clock to prevent type 2 diabetes mellitus?
The circadian clock drives a number of metabolic processes including energy intake, storage and utilization coupled with the sleep/wake cycles. Globally, the increasing prevalence of type 2 diabetes (T2DM) has become a significant international public health concern. In view of the heavy societal burden caused by diabetes, and further, to reduce its growing incidence, it is clearly essential to understand the causes of this disease and to devise more effective strategies for its treatment. Although many factors cause T2DM, this article centers on the role of circadian regulation of metabolism. The correlation between the increased occurrence of T2DM and the ubiquity of modern social pressures such as 24/7 lifestyles as well as nocturnal lighting conditions point strongly to the hypothesis that malfunctioning of circadian controls may be involved in the etiology of the illness. Nocturnal light exposure, unusual timing of food, irregular sleep/wake schedules and traveling between different time zones are some of the factors responsible for improper entrainment of the clock. Recent reports have proposed that strengthening of circadian clock functioning and proper timing of food intake could stabilize glucose homeostasis. This strategy thus represents a chronotherapeutic option for non-pharmaceutical intervention in treating T2DM patients.


Patterns of chemotherapy use and outcomes in advanced non-small cell lung cancer by age in England: A retrospective analysis of the population-based Systemic Anti-Cancer Treatment (SACT) dataset.
We described the patterns of chemotherapy use and outcomes in patients diagnosed with stage III or IV non-small cell lung cancer (NSCLC) by age in England. In this retrospective population-based study, we included 20,716 (62% stage IV) patients with NSCLC diagnosed from 2014 to 2017 treated with chemotherapy. We used the Systemic Anti-Cancer Treatment (SACT) dataset to describe changes in treatment plan and estimated 30 and 90-day mortality rates and median, 6-, and 12-month overall survival (OS) using Kaplan Meier estimator for patients aged <75 and ≥ 75 by stage. Using flexible hazard regression models we assessed the impact of age, stage, treatment intent (stage III), and performance status on survival. Patients aged ≥75 years were less likely to receive two or more regimens, more likely to have their treatment modified because of comorbidities and their doses reduced compared to younger patients. However, early mortality rates and overall survival were similar across ages, apart from the oldest patients with stage III disease. This observational study demonstrates that age is associated with treatment patterns in an older population with advanced NSCLC in England. Although this reflects a pre-immunotherapy period, given the median age of NSCLC patients and increasingly older population, these results suggest older patients (>75 yrs) may benefit from more intense treatments.


Flat, rectangular frequency comb generation with tunable bandwidth and frequency spacing.
The generation of flat, rectangular frequency combs with tunable frequency spacing and bandwidth is demonstrated. Therefore, several lines or sidebands are extracted out of an existing frequency comb, for example a femtosecond fiber laser. Subsequently, these lines are processed via two Mach-Zehnder modulators in order to generate a flat frequency comb with tunable frequency spacing. Optical frequency combs with various spacing and a maximum bandwidth of 260 GHz are generated. However, much higher bandwidth can be reached easily. The overall flatness of the generated combs is within 0.6 dB.


Crystal structure of the CD2-binding domain of CD58 (lymphocyte function-associated antigen 3) at 1.8-A resolution.
The binding of the cell surface molecule CD58 (formerly lymphocyte function-associated antigen 3) to its ligand, CD2, significantly increases the sensitivity of antigen recognition by T cells. This was the first heterophilic cell adhesion interaction to be discovered and is now an important paradigm for analyzing the structural basis of cell-cell recognition. The crystal structure of a CD2-binding chimeric form of CD58, solved to 1.8-A resolution, reveals that the ligand binding domain of CD58 has the expected Ig superfamily V-set topology and shares several of the hitherto unique structural features of CD2, consistent with previous speculation that the genes encoding these molecules arose via duplication of a common precursor. Nevertheless, evidence for considerable divergence of CD2 and CD58 is also implicit in the structures. Mutations that disrupt CD2 binding map to the highly acidic surface of the AGFCC'C" beta-sheet of CD58, which, unexpectedly, lacks marked shape complementarity to the equivalent, rather more basic CD58-binding face of human CD2. The specificity of the very weak interactions of proteins mediating cell-cell recognition may often derive largely from electrostatic complementarity, with shape matching at the protein-protein interface being less exact than for interactions that combine specificity with high affinity, such as those involving antibodies.


PEGylation extends circulation half-life while preserving in vitro and in vivo activity of tissue inhibitor of metalloproteinases-1 (TIMP-1).
Excess proteolytic activity of matrix metalloproteinases (MMPs) contributes to the development of arthritis, cardiovascular diseases and cancer progression, implicating these enzymes as therapeutic targets. While many small molecule inhibitors of MMPs have been developed, clinical uses have been limited, in part by toxicity and off-target effects. Development of the endogenous tissue inhibitors of metalloproteinases (TIMPs) as recombinant biopharmaceuticals represents an alternative therapeutic approach; however, the short plasma half-life of recombinant TIMPs has restricted their potential in this arena. To overcome this limitation, we have modified recombinant human TIMP-1 (rhTIMP-1) by PEGylation on lysine residues. We analyzed a mixture of mono- and di-PEGylated rhTIMP-1 species modified by attachment of 20 kDa mPEG chains (PEG(20K)-TIMP-1), as confirmed by SELDI-TOF mass spectrometry. This preparation retained complete inhibitory activity toward the MMP-3 catalytic domain and partial inhibitory activity toward full length MMP-9. Pharmacokinetic evaluation showed that PEGylation extended the plasma half-life of rhTIMP-1 in mice from 1.1 h to 28 h. In biological assays, PEG(20K)-TIMP-1 inhibited both MMP-dependent cancer cell invasion and tumor cell associated gelatinase activity. Overall these results suggest that PEGylated TIMP-1 exhibits improved potential for development as an anti-cancer recombinant protein therapeutic, and additionally may offer potential for clinical applications in the treatment of other diseases.


Refinement of the use of food and fluid control as motivational tools for macaques used in behavioural neuroscience research: report of a Working Group of the NC3Rs.
This report provides practical guidance on refinement of the use of food and fluid control as motivational tools for macaques used in behavioural neuroscience research. The guidance is based on consideration of the scientific literature and, where data are lacking, expert opinion and professional experience, including that of the members of a Working Group convened by the United Kingdom National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs). The report should be useful to researchers, veterinarians and animal care staff responsible for the welfare of macaques used in food and fluid control protocols, as well as those involved with designing, performing and analysing studies that use these protocols. It should also assist regulatory authorities and members of local ethical review processes or institutional animal care and use committees concerned with evaluating such protocols. The report provides a framework for refinement that can be tailored to meet local requirements. It also identifies data gaps and areas for future research and sets out the Working Group's recommendations on contemporary best practice.


Use of AUDIT, and measures of drinking frequency and patterns to detect associations between alcohol and sexual behaviour in male sex workers in Kenya.
Previous research has linked alcohol use with an increased number of sexual partners, inconsistent condom use and a raised incidence of sexually transmitted infections (STIs). However, alcohol measures have been poorly standardised, with many ill-suited to eliciting, with adequate precision, the relationship between alcohol use and sexual risk behaviour. This study investigates which alcohol indicator--single-item measures of frequency and patterns of drinking ( > = 6 drinks on 1 occasion), or the Alcohol Use Disorders Identification Test (AUDIT)--can detect associations between alcohol use and unsafe sexual behaviour among male sex workers. A cross-sectional survey in 2008 recruited male sex workers who sell sex to men from 65 venues in Mombasa district, Kenya, similar to a 2006 survey. Information was collected on socio-demographics, substance use, sexual behaviour, violence and STI symptoms. Multivariate models examined associations between the three measures of alcohol use and condom use, sexual violence, and penile or anal discharge. The 442 participants reported a median 2 clients/week (IQR = 1-3), with half using condoms consistently in the last 30 days. Of the approximately 70% of men who drink alcohol, half (50.5%) drink two or more times a week. Binge drinking was common (38.9%). As defined by AUDIT, 35% of participants who drink had hazardous drinking, 15% harmful drinking and 21% alcohol dependence. Compared with abstinence, alcohol dependence was associated with inconsistent condom use (AOR = 2.5, 95%CI = 1.3-4.6), penile or anal discharge (AOR = 1.9, 95%CI = 1.0-3.8), and two-fold higher odds of sexual violence (AOR = 2.0, 95%CI = 0.9-4.9). Frequent drinking was associated with inconsistent condom use (AOR = 1.8, 95%CI = 1.1-3.0) and partner number, while binge drinking was only linked with inconsistent condom use (AOR = 1.6, 95%CI = 1.0-2.5). Male sex workers have high levels of hazardous and harmful drinking, and require alcohol-reduction interventions. Compared with indicators of drinking frequency or pattern, the AUDIT measure has stronger associations with inconsistent condom use, STI symptoms and sexual violence. Increased use of the AUDIT tool in future studies may assist in delineating with greater precision the explanatory mechanisms which link alcohol use, drinking contexts, sexual behaviours and HIV transmission.


Correlates of sleep and waking in Drosophila melanogaster.
Drosophila exhibits a circadian rest-activity cycle, but it is not known whether fly rest constitutes sleep or is mere inactivity. It is shown here that, like mammalian sleep, rest in Drosophila is characterized by an increased arousal threshold and is homeostatically regulated independently of the circadian clock. As in mammals, rest is abundant in young flies, is reduced in older flies, and is modulated by stimulants and hypnotics. Several molecular markers modulated by sleep and waking in mammals are modulated by rest and activity in Drosophila, including cytochrome oxidase C, the endoplasmic reticulum chaperone protein BiP, and enzymes implicated in the catabolism of monoamines. Flies lacking one such enzyme, arylalkylamine N-acetyltransferase, show increased rest after rest deprivation. These results implicate the catabolism of monoamines in the regulation of sleep and waking in the fly and suggest that Drosophila may serve as a model system for the genetic dissection of sleep.


Aurora-A kinase maintains the fidelity of early and late mitotic events in HeLa cells.
Aurora-A, a member of the Aurora/Ipl1-related kinase family, is overexpressed in various types of cancer and considered to play critical roles in tumorigenesis. To better understand the pathological effect of Aurora-A activation, it is first necessary to elucidate the physiological functions of Aurora-A. Here, we have investigated the roles of Aurora-A in mitotic progression with the small interfering RNA, antibody microinjection, and time lapse microscopy using human cells. We demonstrated that suppression of Aurora-A by small interfering RNA caused multiple events to fail in mitosis, such as incorrect separation of centriole pairs, misalignment of chromosomes on the metaphase plate, and incomplete cytokinesis. Antibody microinjection of Aurora-A into late G2 cells induced dose-dependent failure in separation of centriole pairs at prophase, indicating that Aurora-A is essential for proper separation of centriole pairs. When we injected anti-Aurora-A antibodies into prometaphase cells that had separated their centriole pairs, chromosomes were severely misaligned on the metaphase plate, indicating that Aurora-A is required for proper movement of chromosomes on the metaphase plate. Furthermore, inhibition of Aurora-A at metaphase by microinjected antibodies prevented cells from completing cytokinesis, suggesting that Aurora-A also has important functions in late mitosis. These results strongly suggest that Aurora-A is essential for many crucial events during mitosis and that the phosphorylation of a series of substrates by Aurora-A at different stages of mitosis may promote diverse critical events in mitosis to maintain chromosome integrity in human cells.


NetOglyc: prediction of mucin type O-glycosylation sites based on sequence context and surface accessibility.
The specificities of the UDP-GalNAc:polypeptide Nacetylgalactosaminyltransferases which link the carbohydrate GalNAc to the side-chain of certain serine and threonine residues in mucin type glycoproteins, are presently unknown. The specificity seems to be modulated by sequence context, secondary structure and surface accessibility. The sequence context of glycosylated threonines was found to differ from that of serine, and the sites were found to cluster. Non-clustered sites had a sequence context different from that of clustered sites. Charged residues were disfavoured at position -1 and +3. A jury of artificial neural networks was trained to recognize the sequence context and surface accessibility of 299 known and verified mucin type O-glycosylation sites extracted from O-GLYCBASE. The cross-validated NetOglyc network system correctly found 83% of the glycosylated and 90% of the non-glycosylated serine and threonine residues in independent test sets, thus proving more accurate than matrix statistics and vector projection methods. Predictions of O-glycosylation sites in the envelope glycoprotein gp120 from the primate lentiviruses HIV-1, HIV-2 and SIV are presented. The most conserved O-glycosylation signals in these evolutionary-related glycoproteins were found in their first hypervariable loop, V1. However, the strain variation for HIV-1 gp120 was significant. A computer server, available through WWW or E-mail, has been developed for prediction of mucin type O-glycosylation sites in proteins based on the amino acid sequence. The server addresses are http://www.cbs.dtu.dk/services/NetOGlyc/ and firstname.lastname@example.com.


Variable angle locking intercarpal fusion system for four-corner arthrodesis: indications and surgical technique.
Four-corner fusion (4CF) is an accepted and regularly performed procedure when managing posttraumatic degenerative disorders in the wrist. This procedure consists of excision of the entire scaphoid in association with midcarpal fusion of the remaining four ulnar carpal bones (hamate, capitate, lunate, and triquetrum). In the majority of cases, the long-term outcome is a functional painless wrist. However, the exact procedure to best achieve a rapid solid bone union of the fusion mass without hardware complications remains controversial. The authors have developed a precise system to ensure precise positioning, firm fixation, and fusion at the midcarpal joint together with an early postoperative recovery, avoiding some of the issues reported with other implants used for 4CF. The described implant is a circular plate accommodating variable angle locking screws as well as compression screws that can firmly fix the plate to the carpal bones. The locking technology produces a very solid construct. A special reaming-distraction-compression guide has also been developed to both countersink the plate on the underlying carpal bone mass and allow distraction of the midcarpal joint for debridement and cancellous bone graft interposition. The features of the implant, its surgical technique, and a relevant case are described.


Novel Nonsense Mutation in SLC39A13 Initially Presenting as Myopathy: Case Report and Review of the Literature.
Myopathies comprise a heterogeneous group of disorders characterized by variable phenotypes. The increasing use of next-generation sequencing allows identification of the causative genes in a much higher percentage of patients with hereditary muscle disorders and also illustrates a considerable degree of overlap with other clinical entities, including connective tissue disorders. Here, we present a 14-year-old German patient who was initially suspected to suffer from myopathy based on his clinical, radiological, and muscle biopsy findings. Exome sequencing revealed a novel homozygous nonsense mutation in the SLC39A13 gene, causative for spondylocheiro dysplastic Ehlers Danlos syndrome (SCD-EDS), suggesting a connective tissue disorder. Including our patient, only 9 affected individuals from 4 families have been described for SCD-EDS so far. The previously reported patients did not show obvious evidence of myopathy, suggesting a broader clinical presentation than originally suspected. We summarize herein the current knowledge on clinical features as well as pathophysiological pathways for this rare connective tissue disease and discuss the high degree of clinical overlap between myopathic and connective tissue disorders.


Relationship of diabetes with cardiovascular disease-related hospitalization rates, length of stay, and charges: analysis by race/ethnicity, age, and sex.
Determine relationship of diabetes with risk of cardiovascular disease hospitalizations and the effect on hospital length of stay and charges. A cross-sectional analysis of Georgia hospital discharge data for 1998 through 2001. Patients hospitalized principally with one of six cardiovascular disease (CVD) conditions (myocardial infarction, ischemic heart disease, cardiac dysrhythmia, heart failure, cerebrovascular events, peripheral vascular disease) were identified in the hospital discharge data. Aggregated CVD-related hospitalization rates, length of stay, and charges were compared by presence of diabetes. Analyses were adjusted for age, sex, and race/ethnicity. A total of 3,900,337 discharges were recorded between 1998 to 2001. Of these, 468,957 discharges (12%) had one of the six selected CVD diagnoses (average age 67 years, average length of stay 4.7 days, average total charge $15,702, 48% women, 76% non-Hispanic Whites, 22% non-Hispanic Blacks, and 1% Hispanics). Diabetes was a concurrent diagnosis in 30% of these CVD-related discharges. CVD hospitalization rates were significantly higher and length of stay and total charges were significantly greater among non-Hispanic Whites and Blacks-but not in Hispanics-with diabetes compared to persons without diabetes. Diabetes had a similar effect on CVD hospitalizations among men and women, but the effect of diabetes was lessened with increasing age. These data suggests that aggressive outpatient modification of metabolic abnormalities in diabetes patients should be attempted to decrease risk of CVD-related hospitalization and lower the economic impact of these combined conditions.


Single-cell isolation by a modular single-cell pipette for RNA-sequencing.
Single-cell transcriptome sequencing highly requires a convenient and reliable method to rapidly isolate a live cell into a specific container such as a PCR tube. Here, we report a modular single-cell pipette (mSCP) consisting of three modular components, a SCP-Tip, an air-displacement pipette (ADP), and ADP-Tips, that can be easily assembled, disassembled, and reassembled. By assembling the SCP-Tip containing a hydrodynamic trap, the mSCP can isolate single cells from 5-10 cells per μL of cell suspension. The mSCP is compatible with microscopic identification of captured single cells to finally achieve 100% single-cell isolation efficiency. The isolated live single cells are in submicroliter volumes and well suitable for single-cell PCR analysis and RNA-sequencing. The mSCP possesses merits of convenience, rapidness, and high efficiency, making it a powerful tool to isolate single cells for transcriptome analysis.


Deconstructing the hedgehog pathway in development and disease.
The Hedgehog (Hh) family of secreted signaling proteins is a master regulator of cell fate determination in metazoans, contributing to both pattern formation during embryonic development and postembryonic tissue homeostasis. In a universally used mode of action, graded distribution of Hh protein induces differential cell fate in a dose-dependent manner in cells that receive Hh. Though much of this pathway has been elucidated from genetically based studies in model organisms, such as Drosophila and mice, the importance of Hh-mediated signaling in humans is clearly evident from malformations and a broad range of cancers that arise when the pathway is corrupted.


Two-dimensional GeP3 as a high capacity electrode material for Li-ion batteries.
Two-dimensional (2D) materials are promising for use in lithium (Li) electrodes due to their high surface ratio. By using density functional theory (DFT) calculations, we investigate the adsorption and diffusion of Li on a newly predicted 2D GeP3 material [Nano Lett., 2016, 17, 1833]. The most favourable adsorption sites for Li are identified, and a semiconducting to metallic transition induced by Li adsorption is found, which indicates excellent electrical conductivity. The GeP3 monolayer has an estimated capacity of 648 mA h g-1, which is almost twice that of commercially used graphite (375 mA h g-1). During full Li intercalation, the GeP3 layer undergoes only 1.2% lattice parameter reduction. Moreover, GeP3 possesses the advantages of a small diffusion barrier (∼0.5 eV) and low average open-circuit voltages (∼0.4 V). Our results highlight a new class of promising anode materials, i.e. 2D phosphide, as potential rechargeable lithium batteries with ultrahigh-capacity, superior ionic conductivity, and low average open-circuit voltage.


[Current Progresses in Developing PET Radiopharmaceuticals for Patients in the Czech Republic].
In Masaryk Memorial Cancer Institute (MMCI), there is a long-running intensive joint effort of the RECAMO project and commercial entities, involving mainly clinical evaluations of state-of-the-art PET radiopharmaceuticals leading to their future availability for Czech physicians and their patients. Recently, the PET tracers [11C]methionine and [18F]fluorocholine, among others, were developed in this cooperation, both of them tracers with high importance for oncologic positron emission tomography diagnostics. [11C]methionine, labeled by carbon-11 with a half-life of 20 min, is a proteosynthesis marker used primarily for brain tumor visualization, whereas [18F]fluorocholine, labeled by fluorine-18 with a half-life of 109 min, is a marker of synthesis of cellular membranes and cell proliferation, its primary use being PET diagnostics of prostate carcinoma. The results of clinical evaluations of both PET radiopharmaceuticals, performed on the basis of parameters agreed and approved beforehand in cooperation of MMCI, RECAMO and the manufacturer of said radiopharmaceuticals, aimed to prove the efficiency and suitability of both compounds for oncologic PET diagnostics for said tumors. In both cases, the radiopharmaceuticals were evaluated in regard to their major use. The obtained results prove the benefits and efficiency of both compounds in PET diagnostics of respective tumors. The results, in the form of clinical evaluation reports, will be used as part of the documentation required for marketing authorization of these compounds for use in the Czech Republic.Key words: positron emission tomography - radiopharmaceuticals - L-methyl-11C-methionine - 18F-fluorocholineThis work was supported by the project MEYS - NPS I - LO1413.The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study.The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers.Submitted: 10. 6. 2016Accepted: 17. 6. 2016.


Three-dimensional True Spine Length: A Novel Technique for Assessing the Outcomes of Scoliosis Surgery.
Current assessment of spine growth for patients undergoing growth friendly surgical treatment for early-onset scoliosis (EOS) is the use of serial, 1-dimensional standard-of-care coronal vertical spine height (SoCVH) measurements. Any growth of the spine out of the coronal plane is missed by the SoCVH, which may underestimate the actual growth of the spine. This study set to validate the novel 3-dimensional true spine length (3DTSL) radiographic measurement technique for measuring growth of EOS patients. 3DTSL measurement accuracy, reliability, and repeatability was assessed using 10 physical model configurations. In addition, interrater and intrarater reliabilities (IRRs) were assessed using interclass coefficient (ICC) analyses of 23 retrospective EOS patient clinical radiographs. 3DTSL measurements were compared with SoCVH measurements. The model assessment showed excellent accuracy with a mean error of 1.2 mm (SD=0.9; range, 0.0 to 3.0) and mean ICC of 0.999.IRR ICCs of the clinical radiographs averaged 0.952 for the 3DTSL and 0.975 for the vertical height whereas IRRs averaged 0.944 and 0.965, respectively (all P<0.001).Mean clinical 3DTSL curve lengths were 193.9 mm (SD=30.0; range, 142.8 to 276.8), whereas the SoCVH averaged 156.1 mm (SD=29.7; range, 74.7 to 207.3). The mean difference between the matched 3DTSL and SoCVH measurements was 37.8 mm (SD=21.4; range, 1.3 to 95.4) and was statistically significant (P<0.0001). On average, the 3DTSL of the measured spines was 124.2% of the measured SoCVH, with a progressive difference as the Cobb or kyphosis angles increased. The novel 3DTSL measurement is accurate, repeatable, and complements the current growth assessments for EOS treatments. Level II-diagnostic study-development of a diagnostic criteria on basis of consecutive patients, with gold standard.


Cytosolic Free N-Glycans Are Retro-Transported Into the Endoplasmic Reticulum in Plant Cells.
During endoplasmic reticulum (ER)-associated degradation, free N-glycans (FNGs) are produced from misfolded nascent glycoproteins via the combination of the cytosolic peptide N-glycanase (cPNGase) and endo-β-N-acetylglucosaminidase (ENGase) in the plant cytosol. The resulting high-mannose type (HMT)-FNGs, which carry one GlcNAc residue at the reducing end (GN1-FNGs), are ubiquitously found in developing plant cells. In a previous study, we found that HMT-FNGs assisted in protein folding and inhibited β-amyloid fibril formation, suggesting a possible biofunction of FNGs involved in the protein folding system. However, whether these HMT-FNGs occur in the ER, an organelle involved in protein folding, remained unclear. On the contrary, we also reported the presence of plant complex type (PCT)-GN1-FNGs, which carry the Lewisa epitope at the non-reducing end, indicating that these FNGs had been fully processed in the Golgi apparatus. Since plant ENGase was active toward HMT-N-glycans but not PCT-N-glycans that carry β1-2xylosyl and/or α1-3 fucosyl residue(s), these PCT-GN1-FNGs did not appear to be produced from fully processed glycoproteins that harbored PCT-N-glycans via ENGase activity. Interestingly, PCT-GN1-FNGs were found in the extracellular space, suggesting that HMT-GN1-FNGs formed in the cytosol might be transported back to the ER and processed in the Golgi apparatus through the protein secretion pathway. As the first step in elucidating the production mechanism of PCT-GN1-FNGs, we analyzed the structures of free oligosaccharides in plant microsomes and proved that HMT-FNGs (Man9-7GlcNAc1 and Man9-8GlcNAc2) could be found in microsomes, which almost consist of the ER compartments.


A genetic algorithm for the automated generation of molecules within constraints.
A genetic algorithm has been designed which generates molecular structures within constraints. The constraints may be any useful function, for example an enzyme active site, a pharmacophore or molecular properties from pattern recognition or rule-induction analyses. The starting point may be random or may utilise known molecules. These are modified to 'grow' into families of structures which, using the evolutionary operators of selection, crossover and mutation evolve to better fit the constraints. The basis of the algorithm is described together with some applications in lead generation, 3D database construction and drug design. Genetic algorithms of this type may have wider applications in chemistry, for example in the design and optimisation of new polymers, materials (e.g. superconducting materials) or synthetic enzymes.


Functional imaging in a low-field, mobile intraoperative magnetic resonance scanner: expanded paradigms.
We previously demonstrated the capability to obtain functional magnetic resonance imaging (MRI) scans of the motor cortex in healthy volunteers using a low-field mobile operating room-based MRI scanner with 0.12-T field strength. Using an expanded (0.15-T), but still mobile, version of this system, our goal was to acquire data showing activation of other areas of functionally important cortex. Five healthy volunteers were scanned with the low-field scanner using finger tapping, hand touch, silent word generation, text listening, and visual stimulation paradigms. The data was analyzed offline using publicly available software. For comparison, the volunteers were then scanned with a 3-T diagnostic MRI scanner. Significant cortical activation was demonstrated on 16 out of 22 images obtained on the operating room-based scanner. Motor activation was most robust, followed by silent word generation, text listening, and hand touch paradigms. The correlation coefficients compared favorably with the images obtained on the 3-T scanner. The signal changes were higher for images obtained with the low-field, mobile scanner compared with those performed with the 3-T diagnostic MRI scanner. Functional MRI scans of multiple cortical areas can be acquired with a low-field strength magnet designed for intraoperative imaging. Further refinement of this technique may allow for the acquisition of true intraoperative functional MRI scans immediately, before, and even during cranial surgery in select patients.


Skeletal consequences of deletion of steroid receptor coactivator-2/transcription intermediary factor-2.
Both estrogen receptor (ER) and peroxisome proliferator-activated receptor gamma (PPARgamma) regulate bone metabolism, and because steroid receptor coactivator (SRC)-2 (TIF-2) enhances ER and PPARgamma activity, we examined the consequences of deletion of SRC-2 on bone using SRC-2 knock out (KO) mice. Loss of SRC-2 resulted in increased bone mass, with SRC-2 KO mice having 80% higher trabecular bone volume as compared with wild type mice. SRC-2 KO mice also had a marked decrease (by 50%) in bone marrow adipocytes. These data suggested that marrow precursor cells in the SRC-2 KO mice may be resistant to the inhibitory effects of endogenous PPARgamma ligands on bone formation. Consistent with this, compared with cultures from wild type mice, marrow stromal cultures from SRC-2 KO mice formed significantly more mineralized nodules (by 3-fold) in the presence of the PPARgamma agonist, rosiglitazone. Using chromatin immunoprecipitation analysis, we demonstrated that in bone marrow stromal cells, loss of SRC-2 leads to destabilization of the transcription complex at the peroxisome proliferator response elements of a number of PPARgamma target genes, resulting in an overall decrease in the expression of adipocyte-related genes and a marked decrease in adipocyte development. Using ovariectomy with or without estrogen replacement, we also demonstrated that SRC-2 KO mice were partially resistant to the skeletal actions of estrogen. Collectively, these findings indicate that loss of SRC-2 leads to partial skeletal resistance to the ER and PPARgamma, but resistance to PPARgamma is dominant, leading to increased bone mass. Modulating SRC-2 action may, thus, represent a novel therapeutic target for osteoporosis.


Characterization of trabecular bone density with ultra-short echo-time MRI at 1.5, 3.0 and 7.0 T--comparison with micro-computed tomography.
The goal of this study was to test the potential of ultra-short echo-time (UTE) MRI at 1.5, 3.0 and 7.0 T for depiction of trabecular bone structure (of the wrist bones), to evaluate whether T2* relaxation times of bone water and parametric maps of T2* of trabecular bone could be obtained at all three field strengths, and to compare the T2* relaxation times with structural parameters obtained from micro-computed tomography (micro-CT) as a reference standard. Ex vivo carpal bones of six wrists were excised en bloc and underwent MRI at 1.5, 3.0 and 7.0 T in a whole-body MR imager using the head coil. A three-dimensional radial fat-suppressed UTE sequence was applied with subsequent acquisitions, with six different echo times TE of 150, 300, 600, 1200, 3500 and 7000 µs. The T2* relaxation time and pixel-wise computed T2* parametric maps were compared with a micro-computed-tomography reference standard providing trabecular bone structural parameters including porosity (defined as the bone-free fraction within a region of interest), trabecular thickness, trabecular separation, trabecular number and fractal dimension (Dk). T2* relaxation curves and parametric maps could be computed from datasets acquired at all field strengths. Mean T2* relaxation times of trabecular bone were 4580 ± 1040 µs at 1.5 T, 2420 ± 560 µs at 3.0 T and 1220 ± 300 µs at 7.0 T, when averaged over all carpal bones. A positive correlation of T2* with trabecular bone porosity and trabecular separation, and a negative correlation of T2* relaxation time with trabecular thickness, trabecular number and fractal dimension, was detected (p < 0.01 for all field strengths and micro-CT parameters). We conclude that UTE MRI may be useful to characterize the structure of trabecular bone, comparable to micro-CT.


Neuronal regulation of cochlear blood flow in the guinea-pig.
1. Previous studies have shown that electrical stimulation (ES) of the guinea-pig cochlea causes a neurally mediated increase in cochlear blood flow (CBF). It is known that the centrifugal neuronal input to the cochlea comes through the perivascular sympathetic plexus from the cervical sympathetic chain and along the vestibular nerve (VN) from the periolivary area of the brainstem. Both of these neuronal systems are distributed topographically in the cochlea. 2. In order to study the neural origins of ES-evoked CBF increase, laser Doppler flowmetry was used to test the following hypotheses. (a) The response is regional, that is, limited to the area of the cochlea stimulated. To test this we performed differential ES of the cochlear turns. CBF was measured from either the third or the first turn. (b) The response is mediated via autonomic receptors within the cochlea. To study this, we applied atropine, succinylcholine and idazoxan locally to the cochlea. (c) The response is influenced by neuronal input via the sympathetic cervical chain (SC) and components of the VN. We stimulated and sectioned the SC, and sectioned the VN, to test this hypothesis. 3. We observed that the CBF response was topographically restricted to the stimulated region. Locally applied muscarinic or nicotinic antagonists (atropine and succinylcholine respectively) did not affect the response. However, local idazoxan (an alpha 2-blocker) eliminated the response. Locally applied adrenaline and SC stimulation modified the dynamic range of the response. SC sectioning enhanced the responsiveness of the cochlear vasculature to ES. The VN section caused a temporary decrease in CBF and elimination of the ES-evoked CBF response. 4. We conclude that the release of dilating agents is topographical with respect to ES current flow, the ES-evoked CBF increase is peripherally mediated via alpha 2-receptors, and the response is influenced by input via the SC. The elimination of the response by VN sectioning proximal to the brainstem indicated that fibres of the VN mediate the CBF increase during direct cochlear ES. The data suggest that these fibres may be the efferent limb of a neural loop involved with the regulation of CBF. Such a system could provide a mechanism for the rapid increase in CBF with organ stress.


The natural history of sciatica associated with disc pathology. A prospective study with clinical and independent radiologic follow-up.
The purpose of this study was to assess the natural history of sciatica due to lumbosacral nerve root compromise and to evaluate the pathomorphologic changes that accompany the natural resolution of the disease. One hundred sixty-five consecutive patients, 114 males and 51 females, with an average age of 41 years (range, 17-72) and an average duration of symptoms of 4.2 months (range, 1-72) presenting with sciatica thought to be due to lumbosacral nerve root compromise were admitted to the study. The cornerstone of treatment was the serial epidural administration of steroid and local anesthetic by the caudal route on an outpatient basis. Lumbar epidural injection or periradicular infiltration at the appropriate level, confirmed under image intensifier, was the next step before considering surgical decompression. An average of three injections (range, 0-8) was received by each patient. Patients underwent clinical examination and computed tomography. Twenty-three patients (14%) underwent surgical decompression. The remainder were clinically assessed at 1 year after presentation, and 111 were rescanned at the appropriate levels. All conservatively managed patients made a satisfactory clinical recovery: average reduction of pain on the visual analog scale was 94% (range, 45-100), and 64 (76%) of the 84 disc herniations and 7 (26%) of the 27 disc bulges showed partial or complete resolution (chi-square = 20.27, P = 0.0001). Thus a high proportion of patients with discogenic sciatica make a satisfactory recovery with aggressive conservative management, and this recovery is accompanied by resolution of disc herniations in a significant number. Only a small proportion of patients needed surgical decompression.


Isolation & antimicrobial susceptibility of Shigella from patients with acute gastroenteritis in Western Nepal.
Shigellae play an important role as a causative organism of acute gastroenteritis, which is a global health problem with significant morbidity and mortality in especially in developing countries. This study was carried out to determine the isolation and pattern of antimicrobial resistance of Shigella in patients with acute gastroenteritis in western Nepal. The study included all patients with acute gastroenteritis who visited a tertiary care hospital at Pokhara, Nepal during a 2-year period (2002-2004). The isolates was confirmed as Shigella by biochemical reaction and slide agglutination test using specific antisera. Antibiotic sensitivity test was determined by agar diffusion method and minimum inhibitory concentration (MIC) of the drugs was detected. Of the 770 stool samples, 83 (10.8%) yielded Shigella. Shigella flexneri caused 56 (67.4%) of the total cases of shigellosis followed by S. dysenteriae 12 (14.5%), S. sonnei 10 (12%) and S. boydii 5 (6%). Of the 83 isolates, 67 (80.7%) showed resistance to various drugs and 62 (74.7%) were resistant to two or more drugs. Resistance to cotrimoxazole was 80.7 per cent followed by tetracycline 74.7 per cent, ampicillin 53.0 per cent, nalidixic acid 31.3 per cent and ciprofloxacin 2.4 per cent. The MIC(50) and MIC(90) values of those drugs were also very high. All isolates were sensitive to cefotaxime and ceftriaxone. The findings of our study suggested that Shigellae was an important etiological agent for acute gastroenteritis, with a high rate of drug resistance and requires constant monitoring in this region.


A 4-way wavelength demultiplexer based on the plasmonic broadband slow wave system.
We propose a broadband slow wave system based on the thin metal-insulator-metal (MIM) graded grating structure composed of two corrugated metal strips with periodic array of grooves on a thin dielectric substrate. The guided spoof surface plasmon polaritons (SSPPs) at different frequencies can be localized at different positions along the ultrathin MIM grating. By introducing specially designed non-corrugated MIM branches with specific lengths at the locations where the EM waves are trapped, the trapped EM waves can be released and propagate along these branches. A 4-way wavelength demultiplexer based on such plasmonic broadband slow wave system is then demonstrated and fabricated. To improve the isolations between different branches at lower frequencies, band-reject filters are inserted at the front of some MIM branches. The measurements and the simulation results have shown very good agreements, which validate the feasibility of the 4-way wavelength demultiplexer.


A five year audit of mammography in a tertiary hospital, North Central Nigeria.
Breast imaging plays a vital role in the multidisciplinary approach to management of breast disease. A baseline data is apt and necessary for collaborative studies. This is a prospective descriptive study carried out between January 2009 and December 2013 at our institution. Patients who had breast imaging were recruited into the study. Film-screen mammograms were acquired with a General Electric (GE) Senographe DMR machine. Breast ultrasound done using an Aloka Prosound SSD-350+ ultrasound machine equipped with linear and curvilinear 7.5-10 MHz transducer. Findings were categorized using the ACR-BIRADS (American college of Radiologists-Breast imaging reporting and data system). Data was collated and analysed using social statistical package (SPSS) version 17. The mammograms of 824 patients were evaluated during this study period. Their age ranged from 40-85 years with a mean age of 50.9 ± 8.1 years. Eight hundred and sixteen (99%) were females and eight (1%) were males. The commonest clinical indication was breast lump (23.9%). The commonest density pattern was BIRADS 2-scattered fibroglandular pattern (43.9%). Mammograms were normal in 266 (32.3%) and positive in 558 (67.7%). The final BIRADS assessment showed BIRADS 0, 1, 2, 3, 4, 5 and 6 constituted 6.6%, 30.1%, 29.7%, 22.2%, 5.9%, 5.0% and 0.5% respectively. The level of awareness of breast cancer is quite high with the positive mammographic yield emphasizing the value of a multidisciplinary approach in the management of breast diseases.


The evolution of H5N1 influenza viruses in ducks in southern China.
The pathogenicity of avian H5N1 influenza viruses to mammals has been evolving since the mid-1980s. Here, we demonstrate that H5N1 influenza viruses, isolated from apparently healthy domestic ducks in mainland China from 1999 through 2002, were becoming progressively more pathogenic for mammals, and we present a hypothesis explaining the mechanism of this evolutionary direction. Twenty-one viruses isolated from apparently healthy ducks in southern China from 1999 through 2002 were confirmed to be H5N1 subtype influenza A viruses. These isolates are antigenically similar to A/Goose/Guangdong/1/96 (H5N1) virus, which was the source of the 1997 Hong Kong "bird flu" hemagglutinin gene, and all are highly pathogenic in chickens. The viruses form four pathotypes on the basis of their replication and lethality in mice. There is a clear temporal pattern in the progressively increasing pathogenicity of these isolates in the mammalian model. Five of six H5N1 isolates tested replicated in inoculated ducks and were shed from trachea or cloaca, but none caused disease signs or death. Phylogenetic analysis of the full genome indicated that most of the viruses are reassortants containing the A/Goose/Guangdong/1/96-like hemagglutinin gene and the other genes from unknown Eurasian avian influenza viruses. This study is a characterization of the H5N1 avian influenza viruses recently circulating in ducks in mainland China. Our findings suggest that immediate action is needed to prevent the transmission of highly pathogenic avian influenza viruses from the apparently healthy ducks into chickens or mammalian hosts.


How genetically heterogeneous is Kabuki syndrome?: MLL2 testing in 116 patients, review and analyses of mutation and phenotypic spectrum.
MLL2 mutations are detected in 55 to 80% of patients with Kabuki syndrome (KS). In 20 to 45% patients with KS, the genetic basis remains unknown, suggesting possible genetic heterogeneity. Here, we present the largest yet reported cohort of 116 patients with KS. We identified MLL2 variants in 74 patients, of which 47 are novel and a majority are truncating. We show that pathogenic missense mutations were commonly located in exon 48. We undertook a systematic facial KS morphology study of patients with KS at our regional dysmorphology meeting. Our data suggest that nearly all patients with typical KS facial features have pathogenic MLL2 mutations, although KS can be phenotypically variable. Furthermore, we show that MLL2 mutation-positive KS patients are more likely to have feeding problems, kidney anomalies, early breast bud development, joint dislocations and palatal malformations in comparison with MLL2 mutation-negative patients. Our work expands the mutation spectrum of MLL2 that may help in better understanding of this molecule, which is important in gene expression, epigenetic control of active chromatin states, embryonic development and cancer. Our analyses of the phenotype indicates that MLL2 mutation-positive and -negative patients differ systematically, and genetic heterogeneity of KS is not as extensive as previously suggested. Moreover, phenotypic variability of KS suggests that MLL2 testing should be considered even in atypical patients.


Modeling protein-protein and protein-peptide complexes: CAPRI 6th edition.
We present the sixth report evaluating the performance of methods for predicting the atomic resolution structures of protein complexes offered as targets to the community-wide initiative on the Critical Assessment of Predicted Interactions (CAPRI). The evaluation is based on a total of 20,670 predicted models for 8 protein-peptide complexes, a novel category of targets in CAPRI, and 12 protein-protein targets in CAPRI prediction Rounds held during the years 2013-2016. For two of the protein-protein targets, the focus was on the prediction of side-chain conformation and positions of interfacial water molecules. Seven of the protein-protein targets were particularly challenging owing to their multicomponent nature, to conformational changes at the binding site, or to a combination of both. Encouragingly, the very large multiprotein complex with the nucleosome was correctly predicted, and correct models were submitted for the protein-peptide targets, but not for some of the challenging protein-protein targets. Models of acceptable quality or better were obtained for 14 of the 20 targets, including medium quality models for 13 targets and high quality models for 8 targets, indicating tangible progress of present-day computational methods in modeling protein complexes with increased accuracy. Our evaluation suggests that the progress stems from better integration of different modeling tools with docking procedures, as well as the use of more sophisticated evolutionary information to score models. Nonetheless, adequate modeling of conformational flexibility in interacting proteins remains an important area with a crucial need for improvement. Proteins 2017; 85:359-377. © 2016 Wiley Periodicals, Inc.


Intramolecular halogen stabilization of silylium ions directs gearing dynamics.
2,6-Bis(2,6-difluorophenyl)phenyldimethylsilanium ion (1a) adopts a ground-state C(2) trigonal-bipyramidal geometry in which fluorines from opposing 2,6-difluorophenyl groups coordinate to the apical positions of the pentavalent silyl cation. Exchange of fluorine at silicon occurs by a disrotatory gearing mechanism wherein one fluorine remains coordinated to silicon throughout the circuit. The "cogwheel" transition state is C(s)-symmetric, with one ring having a dihedral angle of 0 degrees and the other a dihedral angle of 90 degrees. This correlated dynamic process is a function of the coordinating halogen. The chloro derivative (1b) adopts a similar ground-state geometry, but exchange of chlorine at silicon follows a conrotatory process.


The effects of Treponema pallidum on human dendritic cells.
Cell mediated immune responses play a prominent role in syphilis, which is caused by Treponema pallidum. The role of dendritic cells (DC) in the syphilitic infection is not well understood in human. In the present study, we studied interaction of T. pallidum with DC, generated from human peripheral blood mononuclear cells with GM-CSF and IL-4. After adding T. pallidum for 16 hours to immature DC at culture day 7, the change of surface antigens on DC was monitored by flow cytometry, the amount of IL-12 in culture supernatant of DC was measured by ELISA and T cell stimulatory capacity of DC was checked in mixed lymphocyte reaction (MLR). We have observed an efficient phagocytosis of T. pallidum by electron microscopy as early as 2 hours after addition of T. pallidum to DC. Interaction of DC with T. pallidum resulted in increased surface expression of CD83 which was proportionally increased according to the number of T. pallidum. Expressions of CD80, CD86 and HLA-DR on DC were slightly increased. The amount of IL-12 in the culture supernatant of DC was increased (1,099 pg/ml) after the addition of T. pallidum. T. pallidum-infected DC also displayed enhanced T cell stimulatory capacity in MLR. As seen from the above, we observed phagocytosis of T. pallidum by DC as early as 2 hours after addition of T. pallidum to DC and found that T. pallidum can stimulate DC maturation which mean that DC modulate an protective immune response during T. pallidum infection.


Beyond Mars and Venus: The role of gender essentialism in support for gender inequality and backlash.
It has been argued that gender essentialism impedes progress towards greater gender equality. Here we present a new gender essentialism scale (GES), and validate it in two large nationally representative samples from Denmark and Australia. In both samples the GES was highly reliable and predicted lack of support for sex-role egalitarianism and support for gender discrimination, as well as perceived fairness of gender-based treatment in the Australian sample, independently of two established predictors (i.e., social dominance orientation and conservative political orientation). In addition, gender essentialism assessed by the GES moderated some manifestations of the backlash effect: high essentialists were more likely to respond negatively towards a power-seeking female political candidate relative to a male candidate. Given the implications for possible workplace interventions, further work could usefully explore whether gender essentialism moderates other well-established forms of gender bias.


Does collaborative care improve social functioning in adults with depression? The application of the WHO ICF framework and meta-analysis of outcomes.
Collaborative care has proven efficacy in improving symptoms of depression, yet patients value improvements in their social function also. We used the World Health Organisation's International classification of functioning, disability, and health (WHO ICF) to robustly identify measures of social function and explored whether collaborative care interventions improve social functioning using meta-analysis. We performed a secondary data analysis on studies identified from our previous Cochrane review of collaborative care interventions for depression and search update (December 2013). The WHO ICF framework was applied to identify studies that included self-report measures of social functioning. Outcomes were extracted at short-term (6 months) and medium-term (≥7 months) and analysed using random-effects meta-analysis. The relationship between improvements in depression outcomes and improvements in social functioning was also explored using bivarable meta-regression. Eighteen trials were identified that measured social functioning and met our remaining inclusion criteria. Collaborative care was associated with small improvements in social functioning in the short (Standardised Mean Difference, SMD=0.23, 95% confidence interval 0.12 to 0.34) and medium term (SMD=0.19, 95% confidence interval 0.09 to 0.29). Improvements in depressive symptoms were associated with moderate improvements in social function (ß=-0.55, 95% confidence interval -0.82 to -0.28) but cross-sectionally only. The small number of studies (N=18) prevented more complex analyses to explore moderators of social functioning outcomes. Collaborative care improves social functioning but the mechanisms through which this occurs are unknown. Future depression interventions need to consider a person's degree of social function equally alongside their depressive symptoms.


Diurnal variation of pulmonary artery pressure in chronic heart failure.
Variation in pulmonary artery pressure has important consequences for the interpretation of isolated pressure measurements in patients with chronic heart failure. To investigate the nature of diurnal variation in pulmonary artery pressure in chronic heart failure, eight angina-free men (aged 50-72 years) with treated chronic heart failure caused by ischaemic heart disease underwent continuous ambulatory pulmonary artery pressure recording by a transducer tipped catheter. The mean (1 SD) daytime pulmonary artery pressure was 29.6 (5.0) mm Hg systolic and 13.7 (5.6) mm Hg diastolic. The mean change in pressure from day to night was +5.1 (3.2) mm Hg systolic and +3.8 (1.7) mm Hg diastolic; and the mean change from standing to lying +9.3 (2.3) mm Hg systolic and +6.4 (2.1) mm Hg diastolic. In six of the eight patients there was considerable rise in pulmonary artery pressure at night, but in the two patients with the most severe symptoms there was no nocturnal rise. In patients with chronic heart failure, nocturnal pulmonary artery pressure is not determined by postural change alone. But interpretation of isolated pulmonary artery pressure measurements must take the posture of the patient into account.


Role of cytochrome P450 2E1 in the metabolism of acrylamide and acrylonitrile in mice.
Acrylonitrile (AN) and acrylamide (AM) are commonly used in the synthesis of plastics and polymers. In rodents, AM and AN are metabolized to the epoxides glycidamide and cyanoethylene oxide, respectively. The aim of this study was to determine the role of cytochrome P450 in the metabolism of AM and AN in vivo. Wild-type (WT) mice, WT mice pretreated with aminobenzotriazole (ABT, 50 mg/kg ip, 2 h pre-exposure), and mice devoid of cytochrome P450 2E1 (P450 2E1-null) were treated with 50 mg/kg [(13)C]AM po. WT mice and P450 2E1-null mice were treated with 2.5 or 10 mg/kg [(13)C]AN po. Urine was collected for 24 h, and metabolites were characterized using (13)C NMR. WT mice excreted metabolites derived from the epoxides and from direct GSH conjugation with AM or AN. Only metabolites derived from direct GSH conjugation with AM or AN were observed in the urine from ABT-pretreated WT mice and P450 2E1-null mice. On the basis of evaluation of urinary metabolites at these doses, these data suggest that P450 2E1 is possibly the only cytochrome P450 enzyme involved in the metabolism of AM and AN in mice, that inhibiting total P450 activity does not result in new pathways of non-P450 metabolism of AM, and that mice devoid of P450 2E1 do not excrete metabolites of AM or AN that would be produced by oxidation by other cytochrome P450s. P450 2E1-null mice may be an appropriate model for the investigation of the role of oxidative metabolism in the toxicity or carcinogenicity of these compounds.


Homogeneous cationic substitution for two-dimensional layered metal oxide nanosheets via a galvanic exchange reaction.
The galvanic exchange reaction of an exfoliated 2D layered metal oxide nanosheet (NS) with excess substituent metal cations enables the synthesis of a mixed metal oxide 2D NS with controllable cation compositions and physicochemical properties. The reaction of the exfoliated MnO2 NS with Fe2+ or Sn2+ ions at 90 °C induces the uniform galvanic replacement of Mn ions with these substituent ions, whereas the same reaction at 25 °C results in the intercalative restacking of the negatively-charged MnO2 NS with Fe2+ or Sn2+ cations. Upon the galvanic exchange reaction, the highly anisotropic MnO2 2D NS retains its original 2D morphology and layered structure, which is in stark contrast to 0D nanoparticles yielding hollow nanospheres via the galvanic exchange reaction. This observation is attributable to the thin thickness of the 2D NS allowing the simultaneous replacement of all the component surface-exposed metal ions. The resulting substitution of the MnO2 NS with Fe and Sn ions remarkably improves the electrode performance of the carbon-coated derivatives of the MnO2 NS for lithium ion batteries. The present study clearly demonstrates that the galvanic exchange reaction can provide an efficient method not only to tailor cation compositions but also to improve the functionalities of 2D metal oxide NSs and their carbon-coated derivatives.


Structural and molecular modeling features of P2X receptors.
Currently, adenosine 5'-triphosphate (ATP) is recognized as the extracellular messenger that acts through P2 receptors. P2 receptors are divided into two subtypes: P2Y metabotropic receptors and P2X ionotropic receptors, both of which are found in virtually all mammalian cell types studied. Due to the difficulty in studying membrane protein structures by X-ray crystallography or NMR techniques, there is little information about these structures available in the literature. Two structures of the P2X4 receptor in truncated form have been solved by crystallography. Molecular modeling has proven to be an excellent tool for studying ionotropic receptors. Recently, modeling studies carried out on P2X receptors have advanced our knowledge of the P2X receptor structure-function relationships. This review presents a brief history of ion channel structural studies and shows how modeling approaches can be used to address relevant questions about P2X receptors.


[Effect of Aromatherapy on Menopausal Symptoms, Heart Rate Variability, and Sleep Quality in Women].
Studies have shown that aromatherapy improve health problems related to anxiety, depression, heart rate variability (HRV), and sleep. However, the effect of aromatherapy in women who suffer from menopausal syndrome and its specific effects on HRV and sleep quality are unknown. This study designed an aromatherapy intervention and evaluated its effect on menopausal syndrome, HRV, and sleep quality in women. This double-blinded randomized controlled trial was conducted at a medical center hospital. A total of 84 participants who met the study criteria were randomly assigned using permuted block randomization. The experimental group received a 20-min inhalation of essential oil and the control group received a 20-min inhalation of sweet almond oil every night for 4 weeks. Posttest data was collected at 2 weeks after completion of the intervention. Data were collected using the Green Menopausal Symptom Scale, HRV device, and Pittsburgh Sleep Quality Index. After adjusting for age, the results of the generalized estimation equations (GEE) showed that all outcomes were significantly different in both the experimental group and the control group (p < .05) for the interaction effect of group and time, and that the outcome of sleep quality on the 2nd week in the control group was not significantly different (p < .066). This study supports that the 4-week aromatherapy intervention improves menopausal symptoms, the activity of autonomic nervous systems, and sleep quality in women with no adverse side effects. In the future, this intervention may be applied in outpatient departments to promote the health of menopausal women.


Association of over-the-counter pharmaceutical sales with influenza-like-illnesses to patient volume in an urgent care setting.
We studied the association between OTC pharmaceutical sales and volume of patients with influenza-like-illnesses (ILI) at an urgent care center over one year. OTC pharmaceutical sales explain 36% of the variance in the patient volume, and each standard deviation increase is associated with 4.7 more patient visits to the urgent care center (p<0.0001). Cross-correlation function analysis demonstrated that OTC pharmaceutical sales are significantly associated with patient volume during non-flu season (p<0.0001), but only the sales of cough and cold (p<0.0001) and thermometer (p<0.0001) categories were significant during flu season with a lag of two and one days, respectively. Our study is the first study to demonstrate and measure the relationship between OTC pharmaceutical sales and urgent care center patient volume, and presents strong evidence that OTC sales predict urgent care center patient volume year round.


Dementia and risk of adverse warfarin-related events in the nursing home setting.
Little attention has been focused on the safety of medications administered to treat non illnesses in nursing home residents with dementia. It is unclear whether this population is at increased risk of adverse drug events. To test the hypotheses that in nursing home residents with dementia prescribed warfarin have less time in therapeutic range and a higher incidence of nonpreventable and preventable adverse warfarin events compared to nursing home residents without dementia after controlling for facility and patient characteristics. A prospective cohort embedded in a clinical trial of nursing home residents prescribed warfarin in 26 nursing homes in Connecticut was observed for up to 12 months. The primary outcome measures included adverse warfarin events (AWEs) (injuries resulting from warfarin use), potential AWEs (INR [international normalized ratio] >4.5 and management error), and AWE preventability based on physician reviews of medical record abstractions. Potential confounders included nursing home structural characteristics (eg, number of beds and for-profit status), nursing staff time, and nursing home regulatory deficiencies (pharmacy, administrative, quality of care, and all other deficiencies). Multivariable Poisson regression analysis was used to determine the independent association of dementia with potential and preventable AWEs using generalized estimating equations to account for clustering within nursing homes. Residents with dementia had no difference in the number of INR monitoring tests or percentage of days in the therapeutic range, but did have an increased risk of AWEs (adjusted incidence rate ratio [IRR], 1.47; 95% confidence interval [CI], 1.20-1.82), and preventable or potential AWEs (adjusted IRR, 1.36; 95% CI, 1.06-1.76) after adjustment for patient characteristics, nursing home quality, and case mix. Greater nursing staff time was protective for preventable and potential AWEs (adjusted IRR, 0.66; 95% CI, 0.48-0.90) but not for nonpreventable AWEs. A diagnosis of dementia was associated with increased risk of nonpreventable and preventable or potential AWEs. Greater nursing staff time was associated with lower risk of preventable AWEs. These findings have implications for quality-of-care reporting and patient safety.


Mortality following stroke during and after acute care according to neighbourhood deprivation: a disease registry study.
Neighbourhood deprivation has been shown to be inversely associated with mortality 1 month after stroke. Whether this disadvantage begins while patients are still receiving acute care is unclear. We aimed to study mortality after stroke specifically in the period while patients are under acute care and the ensuing period when they are discharged to home or other care settings. Our sample includes 1760 incident strokes (mean age 75, 48% men, 86% ischaemic) identified between 1998 and 2010 by the population-based stroke registry of Dijon (France). We used Cox regression to study all-cause mortality up to 90 days after stroke occurrence. Overall, 284 (16.1%) patients died during the 90 days following stroke. Prior to stroke, risk factors prevalence (eg, high blood pressure and diabetes) and acute care management did not vary across deprivation levels. There was no association between deprivation and mortality while patients were in acute care (HR comparing the highest to the lowest tertiles of deprivation: 1.01, 95% CI 0.71 to 1.43). After discharge, however, age and gender adjusted mortality gradually increased with deprivation (HR 2.08, 95% CI 1.07 to 4.02). This association was not modified when stroke type and severity were accounted for. The gradient of higher poststroke mortality with increasing neighbourhood deprivation was noticeable only after acute hospital discharge. Quality of postacute care and social support are potential determinants of these variations.


PIK3CA mutations in androgen receptor-positive triple negative breast cancer confer sensitivity to the combination of PI3K and androgen receptor inhibitors.
Triple negative breast cancer (TNBC) is a heterogeneous collection of biologically diverse cancers, which contributes to variable clinical outcomes. Previously, we identified a TNBC subtype that has a luminal phenotype and expresses the androgen receptor (AR+). TNBC cells derived from these luminal AR + tumors have high frequency phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) mutations. The purpose of this study was to determine if targeting phosphoinositide 3-kinase (PI3K) alone or in combination with an AR antagonist is effective in AR + TNBC. We determined the frequency of activating PIK3CA mutations in AR + and AR- TNBC clinical cases. Using AR + TNBC cell line and xenograft models we evaluated the effectiveness of PI3K inhibitors, used alone or in combination with an AR antagonist, on tumor cell growth and viability. PIK3CA kinase mutations were highly clonal, more frequent in AR + vs. AR- TNBC (40% vs. 4%), and often associated with concurrent amplification of the PIK3CA locus. PI3K/mTOR inhibitors had an additive growth inhibitory effect when combined with genetic or pharmacological AR targeting in AR + TNBC cells. We also analyzed the combination of bicalutamide +/- the pan-PI3K inhibitor GDC-0941 or the dual PI3K/mTOR inhibitor GDC-0980 in xenograft tumor studies and observed additive effects. While approximately one third of TNBC patients respond to neoadjuvant/adjuvant chemotherapy, recent studies have shown that patients with AR + TNBC are far less likely to benefit from the current standard of care chemotherapy regimens and novel targeted approaches need to be investigated. In this study, we show that activating PIK3CA mutations are enriched in AR + TNBC; and, we show that the growth and viability of AR + TNBC cell line models is significantly reduced after treatment with PI3K inhibitors used in combination with an AR antagonist. These results provide rationale for pre-selection of TNBC patients with a biomarker (AR expression) to investigate the use of AR antagonists in combination with PI3K/mTOR inhibitors.


Bigh3 is upregulated in regenerating zebrafish fin.
Zebrafish is a good model for studying regeneration because of the rapidity with which it occurs. Better understanding of this process may lead in the future to improvement of the regenerating capacity of humans. Signaling factors are the second largest category of genes, regulated during regeneration after the regulators of wound healing. Major developmental signaling pathways play a role in this multistep process, such as Bmp, Fgf, Notch, retinoic acid, Shh, and Wnt. In the present study, we focus on TGF-β-induced genes, bigh3 and bambia. Bigh3 encodes keratoepithelin, a protein first identified as an extracellular matrix protein reported to play a role in cell adhesion, as well as in cornea formation and osteogenesis. The expression of bigh3 in zebrafish fins has previously been reported. Here we demonstrate that tgf-b1 and tgf-b3 mRNA reacted with delay, first showing no regulation at 3 dpa, followed by upregulation at 4 and 5 dpa. Tgf-b1, tgf-2, and tgf-brII mRNA were back to normal levels at 10 dpa. Only tgf-b3 mRNA was still upregulated at that time. Bigh3 mRNA followed the upregulation of tgf-b1, while bambia mRNA behaved similarly to tgf-b2 mRNA. We show that upregulation of bigh3 and bambia mRNA correlated with the process of fin regeneration and regulation of TGF-b signaling, suggesting a new role for these proteins.


Evolution of nonsurgical therapy for colorectal cancer.
The management of colorectal cancer (CRC) has changed considerably in the past 15 years with the introduction of multiple novel active therapeutic agents. Chemotherapy regimens combining a fluoropyrimidine with either oxaliplatin or irinotecan are standard first-line and second-line therapy for advanced and metastatic disease. The first-line use of these combinations produces tumor response rates of approximately 50% and a median overall survival of approximately 20 months. Addition of bevacizumab to first-line treatment and addition of cetuximab to salvage therapy for patients who fail to respond to irinotecan have contributed to further increases in tumor response rates and enhanced progression-free survival. Such approaches have produced only marginal overall survival benefits, however, and entail considerable cost. Adjuvant chemotherapy, delivered after surgical resection of the primary tumor, increases cure rates by approximately 10% for stage III disease and approximately 3-4% for stage II disease. Encouraging reductions in local relapse rates have been observed in patients with early rectal cancer who have undergone chemoradiotherapy, and increasingly complex regimens are currently being explored in phase II clinical trials in an attempt to increase both the operability and long-term local control of CRC. The greater the therapeutic choice, the greater the cost (both financial and in terms of toxicity), thus the keener the clinical community becomes to develop biomarkers to select patient populations who will be most likely to benefit from a specific agent.


Oligodendrocyte lineage cells contribute unique features to Rett syndrome neuropathology.
Mutations in the methyl-CpG binding protein 2 gene, Mecp2, affect primarily the brain and lead to a wide range of neuropsychiatric disorders, most commonly Rett syndrome (RTT). Although the neuropathology of RTT is well understood, the cellular and molecular mechanism(s), which lead to the disease initiation and progression, has yet to be elucidated. RTT was initially attributed only to neuronal dysfunction, but our recent studies and those of others show that RTT is not exclusively neuronal but rather also involves interactions between neurons and glia. Importantly, studies have shown that MeCP2-restored astrocytes and microglia are able to attenuate the disease progression in otherwise MeCP2-null mice. Here we show that another type of glia, oligodendrocytes, and their progenitors are also involved in manifestation of specific RTT symptoms. Mice that lost MeCP2 specifically in the oligodendrocyte lineage cells, although overall normal, were more active and developed severe hindlimb clasping phenotypes. Inversely, restoration of MeCP2 in oligodendrocyte lineage cells, in otherwise MeCP2-null mice, although only mildly prolonging their lifespan, significantly improved the locomotor deficits and hindlimb clasping phenotype, both in male and female mice, and fully restored the body weight in male mice. Finally, we found that the level of some myelin-related proteins was impaired in the MeCP2-null mice. Expression of MeCP2 in oligodendrocytes of these mice only partially restored their expression, suggesting that there is a non-cell-autonomous effect by other cell types in the brains on the expression of myelin-related proteins in oligodendrocytes.


Effectiveness of N-acetylcysteine in Treating Clinical Symptoms of Substance Abuse and Dependence: A Meta-analysis of Randomized Controlled Trials.
Treatment with N-acetylcysteine (NAC) is believed to reduce the clinical symptoms among individuals with substance abuse or dependence. We conducted a meta-analysis of randomized controlled trials to evaluate the effectiveness of NAC in treating substance abuse and dependence. PubMed, EMBASE, ClinicalTrials.gov registry, and the Cochrane Library were searched for trials published before June 2020. A total of 16 trials were analyzed. The treatment effectiveness domains assessed in this study were craving and depressive symptoms, withdrawal syndrome, adverse events, and smoking frequency. Standardized mean difference (SMD), weighted mean difference (WMD), and odds ratio (OR) were used for evaluation where appropriate. A significant decrease in craving symptoms was observed in the NAC treatment group compared with the control group (SMD, -0.67; 95% confidence interval [CI], -1.21 to 0.21). When withdrawal and depressive symptoms were considered as a single domain, the NAC treatment group demonstrated a significantly higher overall improvement than the control group (SMD, -0.35; 95% CI, -0.64 to -0.06). No between-group differences in term of the OR of adverse events (OR, 1.18; 95% CI, 0.68 to 2.06) and a non-significant trend toward reduction in smoking frequency was observed in the NAC treatment group compared with the control group (WMD, -3.09; 95% CI, -6.50 to 0.32). NAC provides certain noticeable benefits in attenuating substance craving and might help alleviate depressive symptoms and withdrawal syndrome. Precautious measures should be considered when using NAC although no difference in adverse effects was found between NAC treatment and control group.