Exploring the interaction of activity limitations with context, systems, community and personal factors in accessing public health care services: A presentation of South African case studies.
There are many factors that influence access to public health services, such as the context people live in, the existing health services, and personal, cultural and community factors. People with disabilities (activity limitations), through their experience of health services, may offer a particular understanding of the performance of the health services, thus exposing health system limitations more clearly than perhaps any other health service user. This article explores how activity limitations interact with factors related to context, systems, community and personal factors in accessing public health care services in South Africa. We present four case studies of people with disabilities from four low-resource diverse contexts in South Africa (rural, semi-rural, farming community and peri-urban) to highlight challenges of access to health services experienced by people with activity limitations in a variety of contexts. One case study of a person with disabilities was chosen from each study setting to build evidence using an intensive qualitative case study methodology to elucidate individual and household experiences of challenges experienced by people with activity limitations when attempting to access public health services. In-depth interviews were used to collect data, using an interview guide. The analysis was conducted in the form of a thematic analysis using the interview topics as a starting point. First, these four case studies demonstrate that equitable access to health services for people with activity limitations is influenced by a complex interplay of a variety of factors for a single individual in a particular context. Secondly, that while problems with access to public health services are experienced by everyone, people with activity limitations are affected in particular ways making them particularly vulnerable in using public health services. The revitalisation of primary health care and the introduction of national health insurance by the Health Department of South Africa open a window of opportunity for policy makers and policy implementers to revisit and address the areas of access to public health services for people with activity limitations.


Nucleus pulposus cellular longevity by telomerase gene therapy.
Nonviral transfection of nucleus pulposus cells with a telomerase expression construct to assess the effects on cellular lifespan, function, karyotypic stability, and transformation properties. To investigate whether telomerase gene therapy can extend the cellular lifespan while retaining functionality of nucleus pulposus cells in a safe manner. Degeneration of the intervertebral disc is an age-related condition in which cells responsible for the maintenance and health of the disc deteriorate with age. Telomerase can extend the cellular lifespan and function of other musculoskeletal tissues, such as the heart, bones, and connective tissues. Therefore, extension of the cellular lifespan and matrix production of intervertebral disc cells may have the potential to delay the degeneration process. Ovine nucleus pulposus cells were lipofectamine transfected in vitro with a human telomerase reverse transcriptase (hTERT) expression construct. Cellular lifespan and matrix transcript levels were determined by cumulative population doublings and real-time RT-PCR, respectively. G1-cell cycle checkpoint, p53 functionality, growth of transfected cells in anchorage-independent or serum starvation conditions, and karyotypic analysis were performed. Transfection was achieved successfully with 340% +/- 7% (mean +/- SD) relative telomerase activity in hTERT-transfected cells. hTERT transfection enabled a 50% extension in mean cellular lifespan and prolonged matrix production of collagen 1 and 2 for more than 282 days. Karyotypic instability was detected but G1-cell cycle checkpoint and p53 was functionally comparable to parental cells with no growth in serum starvation or anchorage-independent conditions. Telomerase can extend the cellular lifespan of nucleus pulposus cells and prolong the production of extracellular matrix. Safety is still unresolved, as karyotypic instability was detected but no loss of contact inhibition, mitogen dependency, or G1-cell cycle checkpoint control was evident.


Histone deacetylase inhibitor MS-275 alone or combined with bortezomib or sorafenib exhibits strong antiproliferative action in human cholangiocarcinoma cells.
To investigate the antiproliferative effect of the histone deacetylase (HDAC) inhibitor MS-275 on cholangiocarcinoma cells alone and in combination with conventional cytostatic drugs (gemcitabine or doxorubicin) or the novel anticancer agents sorafenib or bortezomib. Two human bile duct adenocarcinoma cell lines (EGI-1 and TFK-1) were studied. Crystal violet staining was used for detection of cell number changes. Cytotoxicity was determined by measuring the release of the cytoplasmic enzyme lactate dehydrogenase (LDH). Apoptosis was determined by measuring the enzyme activity of caspase-3. Cell cycle status reflected by the DNA content was detected by flow cytometry. MS-275 treatment potently inhibited the proliferation of EGI-1 and TFK-1 cholangiocarcinoma cells by inducing apoptosis and cell cycle arrest. MS-275-induced apoptosis was characterized by activation of caspase-3, up-regulation of Bax and down-regulation of Bcl-2. Cell cycle was predominantly arrested at the G(1)/S checkpoint, which was associated with induction of the cyclin-dependent kinase inhibitor p21(Waf/CIP1). Furthermore, additive anti-neoplastic effects were observed when MS-275 treatment was combined with gemcitabine or doxorubicin, while combination with the multi-kinase inhibitor sorafenib or the proteasome inhibitor bortezomib resulted in overadditive anti-neoplastic effects. The growth of human cholangiocarcinoma cells can be potently inhibited by MS-275 alone or in combination with conventional cytostatic drugs or new, targeted anticancer agents.


Use of a management plan for treating asthma in an emergency department.
A standardised management protocol has been developed for the assessment and treatment of adults with acute asthma attending an emergency department. The management protocol consists of an assessment sheet for recording essential features of the history and examination findings and a flow diagram with guidelines for initial management that were based on spirometric recordings. The protocol was introduced at Wellington Hospital in 1986. The effect of this intervention was assessed by analysing emergency department records during the three months before and one year after the introduction of the protocol. The use of the assessment sheet improved history taking and led to the increased use of serial measures of airflow obstruction and improved documentation of follow up arrangements. The provision of management guidelines influenced the emphasis of management, including an increased use of corticosteroids intravenously and more frequent use of an additional dose of nebulised bronchodilator. In the light of the initial experience the protocol has been modified and its use either in an emergency department or in general practice is recommended.


Modeled Osteopathic Manipulative Treatments: A Review of Their in Vitro Effects on Fibroblast Tissue Preparations.
A key osteopathic tenet involves the body's ability to self-heal. Osteopathic manipulative treatment (OMT) has been evolved to improve this healing capacity. The authors' in vitro work has focused on modeling 2 common OMT modalities: myofascial release (MFR) and counterstrain. Their studies have evaluated the effects of these modalities on wound healing, cytokine secretion, and muscle repair. The key components of the host response to mechanical forces are fibroblasts, which are the main fascial cells that respond to different types of strain by secreting anti-inflammatory chemicals and growth factors, thus improving wound healing and muscle repair processes. The purpose of this review is to discuss the cellular and molecular mechanisms by which MFR and other OMT modalities work, in particular, the role of strained fibroblasts in inflammation, wound healing, and muscle repair and regeneration. Changing MFR parameters, such as magnitude, duration, direction, and frequency of strain, might uniquely affect the physiologic response of fibroblasts, muscle contraction, and wound healing. If such results are clinically translatable, the mechanisms underlying the clinical outcomes of OMT modalities will be better understood, and these treatments will be more widely accepted as evidence-based, first-line therapies.


Hospital characteristics associated with feeding tube placement in nursing home residents with advanced cognitive impairment.
Tube-feeding is of questionable benefit for nursing home residents with advanced dementia. Approximately two-thirds of US nursing home residents who are tube fed had their feeding tube inserted during an acute care hospitalization. To identify US hospital characteristics associated with higher rates of feeding tube insertion in nursing home residents with advanced cognitive impairment. The sample included nursing home residents aged 66 years or older with advanced cognitive impairment admitted to acute care hospitals between 2000 and 2007. Rate of feeding tube placement was based on a 20% sample of all Medicare Claims files and was assessed in hospitals with at least 30 such admissions during the 8-year period. A multivariable model with the unit of the analysis being the hospital admission identified hospital-level factors independently associated with feeding tube insertion rates, including bed size, ownership, urban location, and medical school affiliation. Measures of each hospital's care practices for all patients with serious chronic illnesses were evaluated, including intensive care unit (ICU) use in the last 6 months of life, the use of hospice services, and the ratio of specialist to primary care physicians. Patient-level characteristics were also considered. Endoscopic or surgical insertion of a gastrostomy tube during a hospitalization. In 2797 acute care hospitals with 280,869 admissions among 163,022 nursing home residents with advanced cognitive impairment, the rate of feeding tube insertion varied from 0 to 38.9 per 100 hospitalizations (mean [SD], 6.5 [5.3]; median [interquartile range], 5.3 [2.6-9.3]). The mean rate of feeding tube insertions per 100 admissions was 7.9 in 2000, decreasing to 6.2 in 2007. Higher insertion rates were associated with the following hospital features: for-profit ownership vs government owned (8.5 vs 5.5 insertions per 100 hospitalizations; adjusted odds ratio [AOR], 1.33; 95% confidence interval [CI], 1.21-1.46), larger size (>310 beds vs <101 beds: 8.0 vs 4.3 insertions per 100 hospitalizations; AOR, 1.48; 95% CI, 1.35-1.63), and greater ICU use in the last 6 months of life (highest vs lowest decile: 10.1 vs 2.9 insertions per 100 hospitalizations; AOR, 2.60; 95% CI, 2.20-3.06). These differences persisted after controlling for patient characteristics. Specialist to primary care ratio and hospice use were weakly or not associated with feeding tube placement. Among nursing home residents with advanced cognitive impairment admitted to acute care hospitals, for-profit ownership, larger hospital size, and greater ICU use was associated with increased rates of feeding tube insertion, even after adjusting for patient-level characteristics.


Alleles versus mutations: Understanding the evolution of genetic architecture requires a molecular perspective on allelic origins.
Perspectives on the role of large-effect quantitative trait loci (QTL) in the evolution of complex traits have shifted back and forth over the past few decades. Different sets of studies have produced contradictory insights on the evolution of genetic architecture. I argue that much of the confusion results from a failure to distinguish mutational and allelic effects, a limitation of using the Fisherian model of adaptive evolution as the lens through which the evolution of adaptive variation is examined. A molecular-based perspective reveals that allelic differences can involve the cumulative effects of many mutations plus intragenic recombination, a model that is supported by extensive empirical evidence. I discuss how different selection regimes could produce very different architectures of allelic effects under a molecular-based model, which may explain conflicting insights on genetic architecture from studies of variation within populations versus between divergently selected populations. I address shortcomings of genome-wide association study (GWAS) practices in light of more suitable models of allelic evolution, and suggest alternate GWAS strategies to generate more valid inferences about genetic architecture. Finally, I discuss how adopting more suitable models of allelic evolution could help redirect research on complex trait evolution toward addressing more meaningful questions in evolutionary biology.


Perceptual and attentional effects on drivers' speed selection at curves.
This paper describes an experiment comparing the relative effectiveness of various types of warnings on drivers' speed selection at curves. The experiment compared three types of curve warnings across three different curve types in a driving simulator. All of the warnings worked reasonably well for severe curves (45 km/h), regardless of demands from a secondary (cell phone) task. For less demanding curves, only those warnings with a strong perceptual component (i.e., implicit cues) were effective in reducing drivers' curve speeds in the presence of the cell phone task. The design implications of these data appear straightforward; curve warnings that contain perceptual components or emphasise the physical features of the curve work best, particularly in cognitively demanding situations. The cell phone task added to driver workload and drivers became less responsive to primary task demands (i.e., speeds were elevated and reaction times were longer).


Natalizumab, Fingolimod and Dimethyl Fumarate Use and Pregnancy-Related Relapse and Disability in Women With Multiple Sclerosis.
To investigate pregnancy-related disease activity in a contemporary multiple sclerosis (MS) cohort. Using data from the MSBase Registry, we included pregnancies conceived after 31 Dec 2010 from women with relapsing-remitting MS or clinically isolated syndrome. Predictors of intrapartum relapse, and postpartum relapse and disability progression were determined by clustered logistic regression or Cox regression analyses. We included 1998 pregnancies from 1619 women with MS. Preconception annualized relapse rate (ARR) was 0.29 (95% CI 0.27-0.32), fell to 0.19 (0.14-0.24) in third trimester, and increased to 0.59 (0.51-0.67) in early postpartum. Among women who used fingolimod or natalizumab, ARR before pregnancy was 0.37 (0.28-0.49) and 0.29 (0.22-0.37), respectively, and increased during pregnancy. Intrapartum ARR decreased with preconception dimethyl fumarate use. ARR spiked after delivery across all DMT groups. Natalizumab continuation into pregnancy reduced the odds of relapse during pregnancy (OR 0.76 per month [0.60-0.95], p=0.017). DMT re-initiation with natalizumab protected against postpartum relapse (HR 0.11 [0.04-0.32], p<0.0001). Breastfeeding women were less likely to relapse (HR 0.61 [0.41-0.91], p=0.016). 5.6% of pregnancies were followed by confirmed disability progression, predicted by higher relapse activity in pregnancy and postpartum. Intrapartum and postpartum relapse probabilities increased among women with MS after natalizumab or fingolimod cessation. In women considered to be at high relapse risk, use of natalizumab before pregnancy and continued up to 34 weeks gestation, with early re-initiation after delivery is an effective option to minimize relapse risks. Strategies of DMT use have to be balanced against potential fetal/neonatal complications.


Transforming big data into computational models for personalized medicine and health care.
Health care systems generate a huge volume of different types of data. Due to the complexity and challenges inherent in studying medical information, it is not yet possible to create a comprehensive model capable of considering all the aspects of health care systems. There are different points of view regarding what the most efficient approaches toward utilization of this data would be. In this paper, we describe the potential role of big data approaches in improving health care systems and review the most common challenges facing the utilization of health care big data.


Modeling and Planning with Macro-Actions in Decentralized POMDPs.
Decentralized partially observable Markov decision processes (Dec-POMDPs) are general models for decentralized multi-agent decision making under uncertainty. However, they typically model a problem at a low level of granularity, where each agent's actions are primitive operations lasting exactly one time step. We address the case where each agent has macro-actions: temporally extended actions that may require different amounts of time to execute. We model macro-actions as options in a Dec-POMDP, focusing on actions that depend only on information directly available to the agent during execution. Therefore, we model systems where coordination decisions only occur at the level of deciding which macro-actions to execute. The core technical difficulty in this setting is that the options chosen by each agent no longer terminate at the same time. We extend three leading Dec-POMDP algorithms for policy generation to the macro-action case, and demonstrate their effectiveness in both standard benchmarks and a multi-robot coordination problem. The results show that our new algorithms retain agent coordination while allowing high-quality solutions to be generated for significantly longer horizons and larger state-spaces than previous Dec-POMDP methods. Furthermore, in the multi-robot domain, we show that, in contrast to most existing methods that are specialized to a particular problem class, our approach can synthesize control policies that exploit opportunities for coordination while balancing uncertainty, sensor information, and information about other agents.


Intermolecular polarizabilities in H2-rare-gas mixtures (H2-He, Ne, Ar, Kr, Xe): insight from collisional isotropic spectral properties.
The report presents results of theoretical and numerical analysis of the electrical properties related to the isotropic part of the polarizability induced by interactions within compounds built up of a hydrogen H2 molecule and a set of noble gas atoms, Rg, ranging from the least massive helium up to the heaviest xenon perturber. The Cartesian components of the collisional polarizabilities of the H2-Rg systems are found by means of the quantum chemistry methods and their dependence on the intermolecular distance is determined. On the basis of these data, the spherical, symmetry adapted components of the trace polarizability are derived in order to provide data sets that are convenient for evaluating collisional spectral profiles of the isotropic polarized part of light scattered by the H2-Rg mixtures. Three independent methods of numerical computing of the spectral intensities are applied at room temperature (295 K). The properties of the roto-translational profiles obtained are discussed in order to determine the role played by contributions corresponding to each of the symmetry adapted parts of the trace polarizability. By spreading the analysis over the collection of the H2-Rg systems, evolution of the spectral properties with the growing masses of the supermolecular compounds can be observed.


Crystallographically limited submicrometer gratings in (100) and (211) silicon.
We demonstrate the fabrication of submicrometer gratings in (100) and (211) Si with periodicities that are appropriate for normal-incidence coupling into Ge-Si waveguides. Gratings of periodicities in the 0.25-0.75microm range were fabricated by a standard holographic lithography technique. Crystallographically preferential wet etching that effectively terminates at the {111} family of planes facilitates the fabrication of symmetric gratings in (100) Si and of blazed gratings in (211) Si. The gratings used in this study are appropriate for the fabrication of integrated first- and second-order, normal-incidence grating couplers at a 1.30-microm wavelength. The blazed gratings show a preferential coupling of more than 80% of the total diffraction into the (+1) diffraction order at normal incidence. The maximum first-order diffraction efficiency obtained with these gratings was approximately 28% in (100) Si and 23% in (211) Si.


Development of an intervention to facilitate implementation and uptake of diabetic retinopathy screening.
'Implementation interventions' refer to methods used to enhance the adoption and implementation of clinical interventions such as diabetic retinopathy screening (DRS). DRS is effective, yet uptake is often suboptimal. Despite most routine management taking place in primary care and the central role of health care professionals (HCP) in referring to DRS, few interventions have been developed for primary care. We aimed to develop a multifaceted intervention targeting both professionals and patients to improve DRS uptake as an example of a systematic development process combining theory, stakeholder involvement, and evidence. First, we identified target behaviours through an audit in primary care of screening attendance. Second, we interviewed patients (n = 47) and HCP (n = 30), to identify determinants of uptake using the Theoretical Domains Framework, mapping these to behaviour change techniques (BCTs) to develop intervention content. Thirdly, we conducted semi-structured consensus groups with stakeholders, specifically users of the intervention, i.e. patients (n = 15) and HCPs (n = 16), regarding the feasibility, acceptability, and local relevance of selected BCTs and potential delivery modes. We consulted representatives from the national DRS programme to check intervention 'fit' with existing processes. We applied the APEASE criteria (affordability, practicability, effectiveness, acceptability, side effects, and equity) to select the final intervention components, drawing on findings from the previous steps, and a rapid evidence review of operationalised BCT effectiveness. We identified potentially modifiable target behaviours at the patient (consent, attendance) and professional (registration) level. Patient barriers to consent/attendance included confusion between screening and routine eye checks, and fear of a negative result. Enablers included a recommendation from friends/family or professionals and recognising screening importance. Professional barriers to registration included the time to register patients and a lack of readily available information on uptake in their local area/practice. Most operationalised BCTs were acceptable to patients and HCPs while the response to feasibility varied. After considering APEASE, the core intervention, incorporating a range of BCTs, involved audit/feedback, electronic prompts targeting professionals, HCP-endorsed reminders (face-to-face, by phone and letter), and an information leaflet for patients. Using the example of an intervention to improve DRS uptake, this study illustrates an approach to integrate theory with user involvement. This process highlighted tensions between theory-informed and stakeholder suggestions, and the need to apply the Theoretical Domains Framework (TDF)/BCT structure flexibly. The final intervention draws on the trusted professional-patient relationship, leveraging existing services to enhance implementation of the DRS programme. Intervention feasibility in primary care will be evaluated in a randomised cluster pilot trial.


Familial aggregation of myopia in the Tehran eye study: estimation of the sibling and parent offspring recurrence risk ratios.
To determine the potential influence of genetic factors on the prevalence of myopia in Tehran. Of 6497 citizens of Tehran sampled from 160 clusters using stratified random cluster sampling, 4565 (70.3%) participated in the study and were referred to a clinic for an extensive eye examination and interview. These were from 1259 nuclear families with the average size of 3.6. Refraction data obtained from 3321 participants aged 16 years and over are presented. Three definitions of myopia, as the spherical equivalent of -0.5, -1, and -2 diopters or less, were used. Familial aggregation of myopia was evaluated with odds ratios and recurrence risk ratios (lambda(R)) using a multiple logistic regression with generalised estimating equations (GEE), adjusted for age, sex, height, and education. Multivariate analyses showed a strong familial aggregation of myopia among siblings (lambda(R) ranging from 2.09 to 3.86) and parent-offspring pairs (lambda(R) from 1.82 to 3.81) adjusted for age, sex, height, and education. The aggregation increased with higher myopia thresholds and with the use of cycloplegic refraction. The odds ratios for spouse pairs were not significantly different from 1.0. The association of myopia with sex, height, and education (and not age) remained significant in the final GEE2 model. The findings indicate a relatively high degree of familial aggregation of myopia in the Tehran population, independent of age, sex, height, and education. This residual aggregation may be a result of heredity or of an unmeasured common environmental effect.


HIV testing and care in Burkina Faso, Kenya, Malawi and Uganda: ethics on the ground.
The ethical discourse about HIV testing has undergone a profound transformation in recent years. The greater availability of antiretroviral therapy (ART) has led to a global scaling up of HIV testing and counseling as a gateway to prevention, treatment and care. In response, critics raised important ethical questions, including: How do different testing policies and practices undermine or strengthen informed consent and medical confidentiality? How well do different modalities of testing provide benefits that outweigh risks of harm? To what degree do current testing policies and programs provide equitable access to HIV services? And finally, what lessons have been learned from the field about how to improve the delivery of HIV services to achieve public health objectives and protections for human rights? This article reviews the empirical evidence that has emerged to answer these questions, from four sub-Saharan African countries, namely: Burkina Faso, Kenya, Malawi and Uganda. Expanding access to treatment and prevention in these four countries has made the biomedical benefits of HIV testing increasingly clear. But serious challenges remain with regard to protecting human rights, informed consent and ensuring linkages to care. Policy makers and practitioners are grappling with difficult ethical issues, including how to protect confidentiality, how to strengthen linkages to care, and how to provide equitable access to services, especially for most at risk populations, including men who have sex with men. The most salient policy questions about HIV testing in these countries no longer address whether to scale up routine PITC (and other strategies), but how. Instead, individuals, health care providers and policy makers are struggling with a host of difficult ethical questions about how to protect rights, maximize benefits, and mitigate risks in the face of resource scarcity.


The effects of culture conditions on the glycosylation of secreted human placental alkaline phosphatase produced in Chinese hamster ovary cells.
The effects of different culture conditions, suspension and microcarrier culture and temperature reduction on the structures of N-linked glycans attached to secreted human placental alkaline phosphatase (SEAP) were investigated for CHO cells grown in a controlled bioreactor. Both mass spectrometry and anion-exchange chromatography were used to probe the N-linked glycan structures and distribution. Complex-type glycans were the dominant structures with small amounts of high mannose glycans observed in suspension and reduced temperature cultures. Biantennary glycans were the most common structures detected by mass spectrometry, but triantennary and tetraantennary forms were also detected. The amount of sialic acid present was relatively low, approximately 0.4 mol sialic acid/mol SEAP for suspension cultures. Microcarrier cultures exhibited a decrease in productivity compared with suspension culture due to a decrease in both maximum viable cell density (15-20%) and specific productivity (30-50%). In contrast, a biphasic suspension culture in which the temperature was reduced at the beginning of the stationary phase from 37 to 33 degrees C, showed a 7% increase in maximum viable cell density, a 62% increase in integrated viable cell density, and a 133% increase in specific productivity, leading to greater than threefold increase in total productivity. Both microcarrier and reduced temperature cultures showed increased sialylation and decreased fucosylation when compared to suspension culture. Our results highlight the importance of glycoform analysis after process modification as even subtle changes (e.g., changing from one microcarrier to another) may affect glycan distributions.


Blue and grey water footprint of textile industry in China.
Water footprint (WF) is a newly developed idea that indicates impacts of freshwater appropriation and wastewater discharge. The textile industry is one of the oldest, longest and most complicated industrial chains in the world's manufacturing industries. However, the textile industry is also water intensive. In this paper, we applied a bottom-up approach to estimate the direct blue water footprint (WFdir,blue) and direct grey water footprint (WFdir,grey) of China's textile industry at sector level based on WF methodology. The results showed that WFdir,blue of China's textile industry had an increasing trend from 2001 to 2010. The annual WFdir,blue surpassed 0.92 Gm(3)/yr (giga cubic meter a year) since 2004 and rose to peak value of 1.09 Gm(3)/yr in 2007. The original and residuary WFdir,grey (both were calculated based on the concentration of chemical oxygen demand (CODCr)) of China's textile industry had a similar variation trend with that of WFdir,blue. Among the three sub-sectors of China's textile industry, the manufacture of textiles sector's annual WFdir,blue and WFdir,grey were much larger than those of the manufacture of textile wearing apparel, footware and caps sector and the manufacture of chemical fibers sector. The intensities of WFdir,blue and WF(res)dir,grey of China's textile industry were year by year decreasing through the efforts of issuing restriction policies on freshwater use and wastewater generation and discharge, and popularization of water saving and wastewater treatment technologies.


The Impact of COVID-19 Quarantine on Patients With Dementia and Family Caregivers: A Nation-Wide Survey.
Previous studies showed that quarantine for pandemic diseases is associated with several psychological and medical effects. The consequences of quarantine for COVID-19 pandemic in patients with dementia are unknown. We investigated the clinical changes in patients with Alzheimer's disease and other dementias, and evaluated caregivers' distress during COVID-19 quarantine. The study involved 87 Italian Dementia Centers. Patients with Alzheimer's Disease (AD), Dementia with Lewy Bodies (DLB), Frontotemporal Dementia (FTD), and Vascular Dementia (VD) were eligible for the study. Family caregivers of patients with dementia were interviewed by phone in April 2020, 45 days after quarantine declaration. Main outcomes were patients' changes in cognitive, behavioral, and motor symptoms. Secondary outcomes were effects on caregivers' psychological features. 4913 patients (2934 females, 1979 males) fulfilled the inclusion criteria. Caregivers reported a worsening in cognitive functions in 55.1% of patients, mainly in subjects with DLB and AD. Aggravation of behavioral symptoms was observed in 51.9% of patients. In logistic regression analysis, previous physical independence was associated with both cognitive and behavioral worsening (odds ratio 1.85 [95% CI 1.42-2.39], 1.84 [95% CI 1.43-2.38], respectively). On the contrary, pandemic awareness was a protective factor for the worsening of cognitive and behavioral symptoms (odds ratio 0.74 [95% CI 0.65-0.85]; and 0.72 [95% CI 0.63-0.82], respectively). Approximately 25.9% of patients showed the onset of new behavioral symptoms. A worsening in motor function was reported by 36.7% of patients. Finally, caregivers reported a high increase in anxiety, depression, and distress. Our study shows that quarantine for COVID-19 is associated with an acute worsening of clinical symptoms in patients with dementia as well as increase of caregivers' burden. Our findings emphasize the importance to implement new strategies to mitigate the effects of quarantine in patients with dementia.


Myocardial intracellular pH in a perfused rainbow trout heart during extracellular acidosis in the presence and absence of adrenaline.
Myocardial intracellular pH was measured in a perfused rainbow trout, Salmo gairdneri, with DMO (5,5-dimethyl-2,4-oxazlidinedione), to test the hypothesis that catecholamines promote active regulation of myocardial pH in order to protect contractility during a respiratory acidosis comparable to that observed after exercise. Under control conditions (extracellular pH = 8.0; PCO2 = 2 Torr), myocardial pH was 7.53 +/- 0.01 (N = 5). Acidosis (extracellular pH = 7.45; PCO2 = 8.6 Torr) reduced contractility, mechanical efficiency and intracellular pH (7.25 +/- 0.04), but did not affect myocardial O2 consumption. The addition of 0.5 mumol l-1 adrenaline during extracellular acidosis prevented the loss of contractility, restored mechanical efficiency, but did not change intracellular pH significantly. Thus, adrenaline enabled cardiac contractility to recover, without intracellular pH regulation, possibly by modulation of sarcolemmal calcium changes. The absence of a myocardial acidosis after exercise in vivo is discussed with respect to possible intracellular pH regulation via lactate uptake and metabolism.


Cannabinoids in the treatment of pain and spasticity in multiple sclerosis.
There is a large amount of evidence to support the view that the psychoactive ingredient in cannabis, delta9-tetrahydrocannabinol (delta9-THC), and cannabinoids in general, can reduce muscle spasticity and pain under some circumstances. Cannabinoid (CB1) receptors in the CNS appear to mediate both of these effects and endogenous cannabinoids may fulfil these functions to some extent under normal circumstances. However, in the context of multiple sclerosis (MS), it is still questionable whether cannabinoids are superior to existing, conventional medicationsfor the treatment of spasticity and pain. In the case of spasticity, there are too few controlled clinical trials to draw any reliable conclusion at this stage. In the case of pain, most of the available trials suggest that cannabinoids are not superior to existing treatments; however, few trials have examined chronic pain syndromes that are relevant to MS. Whether or not cannabinoids do have therapeutic potential in the treatment of MS, a further issue will be whether synthetic cannabinoids should be used in preference to cannabis itself. Smoking cannabis is associated with significant risks of lung cancer and other respiratory dysfunction. Furthermore, delta9-THC, as a broad-spectrum cannabinoid receptor agonist, will activate both CB1 and CB2 receptors. Synthetic cannabinoids, which target specific cannabinoid receptor subtypes in specific parts of the CNS, are likely to be of more therapeutic use than delta9-THC itself. If rapid absorption is necessary, such synthetic drugs could be delivered via aerosol formulations.


Neurophysiological Evaluation of Students' Experience during Remote and Face-to-Face Lessons: A Case Study at Driving School.
Nowadays, fostered by technological progress and contextual circumstances such as the economic crisis and pandemic restrictions, remote education is experiencing growing deployment. However, this growth has generated widespread doubts about the actual effectiveness of remote/online learning compared to face-to-face education. The present study was aimed at comparing face-to-face and remote education through a multimodal neurophysiological approach. It involved forty students at a driving school, in a real classroom, experiencing both modalities. Wearable devices to measure brain, ocular, heart and sweating activities were employed in order to analyse the students' neurophysiological signals to obtain insights into the cognitive dimension. In particular, four parameters were considered: the Eye Blink Rate, the Heart Rate and its Variability and the Skin Conductance Level. In addition, the students filled out a questionnaire at the end to obtain an explicit measure of their learning performance. Data analysis showed higher cognitive activity, in terms of attention and mental engagement, in the in-presence setting compared to the remote modality. On the other hand, students in the remote class felt more stressed, particularly during the first part of the lesson. The analysis of questionnaires demonstrated worse performance for the remote group, thus suggesting a common "disengaging" behaviour when attending remote courses, thus undermining their effectiveness. In conclusion, neuroscientific tools could help to obtain insights into mental concerns, often "blind", such as decreasing attention and increasing stress, as well as their dynamics during the lesson itself, thus allowing the definition of proper countermeasures to emerging issues when introducing new practices into daily life.


Validity of the Arabic version of the eating disorders inventory (EDI).
We aimed to validate the Eating Disorders Inventory (EDI) in Arabic. Subjects were chosen randomly from female school students. Only healthy Saudi students were included. They were asked to fill the EDI and undergo a semi-structured interview by a psychiatrist who was unaware of the EDI scores. Of 146 students approached 12 did not meet our criteria and were excluded; 16 other students were excluded for incomplete responses. The difference between the two diagnostic methods in the proportion of caseness was statistically significantly (P < 0.0001). The scores on Perfectionism, Maturity Fears and Interoceptive Awareness were significantly higher in the Saudi students compared with Canadians (P < 0.05). The EDI-DT subscale has a sensitivity of 100% and a specificity of 85%. Despite its low positive predictive value of 5%, it may be useful for screening large non-clinical groups for eating disorders.


Terahertz near-field imaging using subwavelength plasmonic apertures and a quantum cascade laser source.
The first demonstration, to our knowledge, of near-field imaging using subwavelength plasmonic apertures with a terahertz quantum cascade laser source is presented. "Bull's-eye" apertures, featuring subwavelength circular apertures flanked by periodic annular corrugations were created using a novel fabrication method. A fivefold increase in intensity was observed for plasmonic apertures over plain apertures of the same diameter. Detailed studies of the transmitted beam profiles were undertaken for apertures with both planarized and corrugated exit facets, with the former producing spatially uniform intensity profiles and subwavelength spatial resolution. Finally, a proof-of-concept imaging experiment is presented, where an inhomogeneous pharmaceutical drug coating is investigated.


Functional and structural roles of the glutathione-binding residues in maize (Zea mays) glutathione S-transferase I.
The isoenzyme glutathione S-transferase (GST) I from maize (Zea mays) was cloned and expressed in Escherichia coli, and its catalytic mechanism was investigated by site-directed mutagenesis and dynamic studies. The results showed that the enzyme promotes proton dissociation from the GSH thiol and creates a thiolate anion with high nucleophilic reactivity by lowering the pK(a) of the thiol from 8.7 to 6.2. Steady-state kinetics fit well to a rapid equilibrium, random sequential Bi Bi mechanism, with intrasubunit modulation between the GSH binding site (G-site) and the electrophile binding site (H-site). The rate-limiting step of the reaction is viscosity-dependent, and thermodynamic data suggest that product release is rate-limiting. Five residues of GST I (Ser(11), His(40), Lys(41), Gln(53) and Ser(67)), which are located in the G-site, were individually replaced with alanine and their structural and functional roles in the 1-chloro-2,4-dinitrobenzene (CDNB) conjugation reaction were investigated. On the basis of steady-state kinetics, difference spectroscopy and limited proteolysis studies it is concluded that these residues: (1) contribute to the affinity of the G-site for GSH, as they are involved in side-chain interaction with GSH; (2) influence GSH thiol ionization, and thus its reactivity; (3) participate in k(cat) regulation by affecting the rate-limiting step of the reaction; and (4) in the cases of His(40), Lys(41) and Gln(53) play an important role in the structural integrity of, and probably in the flexibility of, the highly mobile short 3(10)-helical segment of alpha-helix 2 (residues 35-46), as shown by limited proteolysis experiments. These structural perturbations are probably transmitted to the H-site through changes in Phe(35) conformation. This accounts for the modulation of K(CDNB)(m) by His(40), Lys(41) and Gln(53), and also for the intrasubunit communication between the G- and H-sites. Computer simulations using CONCOORD were applied to maize GST I monomer and dimer structures, each with bound lactoylglutathione, and the results were analysed by the essential dynamics technique. Differences in dynamics were found between the monomer and the dimer simulations showing the importance of using the whole structure in dynamic analysis. The results obtained confirm that the short 3(10)-helical segment of alpha-helix 2 (residues 35-46) undergoes the most significant structural rearrangements. These rearrangements are discussed in terms of enzyme catalytic mechanism.


Predictive Ability of Laboratory Indices for Liver Fibrosis in Patients with Chronic Hepatitis C after the Eradication of Hepatitis C Virus.
Liver fibrosis remains an important risk factor for hepatocarcinogenesis in patients with chronic hepatitis C even after the eradication of hepatitis C virus (HCV). However, it is difficult to estimate liver fibrosis based on liver biopsy after the eradication of HCV. We investigated the ability of laboratory indices to predict liver fibrosis in patients with sustained virologic response (SVR) to antiviral therapy. Three laboratory liver fibrosis indices (aspartate aminotransferase-platelet ratio index (APRI), FIB-4 index, and Forns index) were calculated based on data at the time of initial pretreatment liver biopsy and at second liver biopsy performed approximately 5 years after SVR in 115 patients who underwent serial liver biopsies. The indices at the time of initial biopsy were compared to histological degree of liver fibrosis in initial biopsy, and laboratory indices at the time of second liver biopsy were compared to the degree of fibrosis in second biopsy. In both comparisons, there were significant increases in all 3 indices with the increase of liver fibrosis grade as assessed in liver biopsy specimens. All 3 indices at the time of second biopsy were able to predict moderate to advanced (METAVIR score F2-4) and advanced (F3-4) fibrosis on liver biopsy, with the area under the receiver-operating characteristics curve >0.8 and the accuracy >70%. All 3 laboratory indices of fibrosis accurately reflected liver fibrosis in patients with SVR for 5 years despite the normalization of serum liver transaminase activity and the lack of liver inflammation.


Murine cytomegalovirus is regulated by a discrete subset of natural killer cells reactive with monoclonal antibody to Ly49H.
Antiviral roles of natural killer (NK) cell subsets were examined in C57BL/6 mice infected with murine cytomegalovirus (MCMV) and other viruses, including lymphocytic choriomeningitis virus (LCMV), vaccinia virus (VV), and mouse hepatitis virus (MHV). Each virus vigorously induced an NK cell infiltrate into the peritoneal cavity and liver, causing some redistributions of NK cell subsets defined by monoclonal antibody (mAb) directed against Ly49A, C/I, D, and G2. Striking results were seen with a mAb (1F8) reactive with the positively signaling molecule Ly49H, present in MCMV-resistant C57BL/6 mice. mAb 1F8 also stains Ly49 C and I, but exclusion of those reactivities with mAb 5E6, which recognizes Ly49 C and I, indicated that Ly49H(+) cells infiltrated the peritoneal cavity and liver and were particularly effective at synthesizing interferon gamma. Depletion of 1F8(+) but not 5E6(+) cells in vivo by mAb injections enhanced MCMV titers by 20-1,000-fold in the spleen and approximately fivefold in the liver. Titers of LCMV or VV were not enhanced. These anti-MCMV effects were attributed to prototypical NK1.1(+)CD3(-) NK cells and not to NK1.1(+)CD3(+) "NK/T" cells. This is the first evidence that control of a virus infection in vivo is mediated by a distinct NK cell subset.


Data-mining of medication records to improve asthma management.
To use community pharmacy medication records to identify patients whose asthma may not be well managed and then implement and evaluate a multidisciplinary educational intervention to improve asthma management. We used a multisite controlled study design. Forty-two pharmacies throughout Tasmania ran a software application that "data-mined" medication records, generating a list of patients who had received three or more canisters of inhaled short-acting beta(2)-agonists in the preceding 6 months. The patients identified were allocated to an intervention or control group. Pre-intervention data were collected for the period May to November 2006 and post-intervention data for the period December 2006 to May 2007. Intervention patients were contacted by the community pharmacist via mail, and were sent educational material and a letter encouraging them to see their general practitioner for an asthma management review. Pharmacists were blinded to the control patients' identities until the end of the post-intervention period. Dispensing ratio of preventer medication (inhaled corticosteroids [ICSs]) to reliever medication (inhaled short-acting beta(2)-agonists). Thirty-five pharmacies completed the study, providing 702 intervention and 849 control patients. The intervention resulted in a threefold increase in the preventer-to-reliever ratio in the intervention group compared with the control group (P < 0.01) and a higher proportion of patients in the intervention group using ICS therapy than in the control group (P < 0.01). Community pharmacy medication records can be effectively used to identify patients with suboptimal asthma management, who can then be referred to their GP for review. The intervention should be trialled on a national scale to determine the effects on clinical, social, emotional and economic outcomes for people in the Australian community, with a longer follow-up to determine sustainability of the improvements noted.


Developmental profile of the aberrant dopamine D2 receptor response in striatal cholinergic interneurons in DYT1 dystonia.
DYT1 dystonia, a severe form of genetically determined human dystonia, exhibits reduced penetrance among carriers and begins usually during adolescence. The reasons for such age dependence and variability remain unclear. We characterized the alterations in D2 dopamine receptor (D2R) signalling in striatal cholinergic interneurons at different ages in mice overexpressing human mutant torsinA (hMT). An abnormal excitatory response to the D2R agonist quinpirole was recorded at postnatal day 14, consisting of a membrane depolarization coupled to an increase in spiking frequency, and persisted unchanged at 3 and 9 months in hMT mice, compared to mice expressing wild-type human torsinA and non-transgenic mice. This response was blocked by the D2R antagonist sulpiride and depended upon G-proteins, as it was prevented by intrapipette GDP-β-S. Patch-clamp recordings from dissociated interneurons revealed a significant increase in the Cav2.2-mediated current fraction at all ages examined. Consistently, chelation of intracellular calcium abolished the paradoxical response to quinpirole. Finally, no gross morphological changes were observed during development. These results suggest that an imbalanced striatal dopaminergic/cholinergic signaling occurs early in DYT1 dystonia and persists along development, representing a susceptibility factor for symptom generation.


QTL-seq for rapid identification of candidate genes for 100-seed weight and root/total plant dry weight ratio under rainfed conditions in chickpea.
Terminal drought is a major constraint to chickpea productivity. Two component traits responsible for reduction in yield under drought stress include reduction in seeds size and root length/root density. QTL-seq approach, therefore, was used to identify candidate genomic regions for 100-seed weight (100SDW) and total dry root weight to total plant dry weight ratio (RTR) under rainfed conditions. Genomewide SNP profiling of extreme phenotypic bulks from the ICC 4958 × ICC 1882 population identified two significant genomic regions, one on CaLG01 (1.08 Mb) and another on CaLG04 (2.7 Mb) linkage groups for 100SDW. Similarly, one significant genomic region on CaLG04 (1.10 Mb) was identified for RTR. Comprehensive analysis revealed four and five putative candidate genes associated with 100SDW and RTR, respectively. Subsequently, two genes (Ca_04364 and Ca_04607) for 100SDW and one gene (Ca_04586) for RTR were validated using CAPS/dCAPS markers. Identified candidate genomic regions and genes may be useful for molecular breeding for chickpea improvement.


Posterior fossa tumor with distinct choroid plexus papilloma and ependymoma components.
Synchronous occurrence of multiple primary central nervous system (CNS) tumors of different histological types is uncommon in patients without radiation history or genetic syndrome association. We herein report a sporadic case of posterior fossa tumor with synchronous choroid plexus papilloma (CPP) and ependymoma (EP) components. A 7-year-old girl with no significant past or familial medical history presented with 2 years of migraine type headaches. Brain magnetic resonance imaging showed a cystic mass with a mural enhancing nodule centered within the cerebellar vermis. The patient underwent gross total resection of the tumor. Histologic examination showed a tumor with two distinct components. The predominant component demonstrated classic morphological and immunohistochemical characteristics of choroid plexus papilloma CPP, WHO grade I. However, there were a few discrete foci, where tumor cells showed architectural, cytological, and immunohistochemical features characteristic of an ependymoma, WHO grade II. In addition, there was exuberant piloid gliosis secondary to infiltration of the CPP component into the adjacent brain parenchyma. Followup brain imaging at 14 months after surgery showed no evidence of residual or recurrent tumor. To the best of our knowledge, this is the first reported case of synchronous CPP and EP in the posterior fossa.


Psychosocial factors at work in relation to back and limb disorders.
This study was a 10-year follow-up of the associations between work content, work control, social relationships at work, mental overstrain, physical work load, and musculoskeletal morbidity in the neck, shoulder and upper limb region, the low back, and the lower limbs among workers in the metal industry. A sample of 902 blue- and white-collar employees were studied. Measurements were made twice at the a 10-year interval by questionnaire and clinical examination. At the beginning of the study, mental overstrain showed several associations with both the symptoms and the findings. The other psychosocial factors were the most consistently associated with the symptoms in the region of the neck, shoulder, and upper limbs and also in the lower-limb region among the middle-aged men. Prospectively, the social relations and the work content scores predicted the change in several morbidity scores. The associations were independent of physical work load. Work-related psychosocial factors were associated with, and predicted, the change in the occurrence of musculoskeletal disorders when age, gender, social class, and physical work load were controlled for.


Canadian Consensus for Biomarker Testing and Treatment of TRK Fusion Cancer in Adults.
The tyrosine receptor kinase (TRK) inhibitors larotrectinib and entrectinib were recently approved in Canada for the treatment of solid tumours harbouring neurotrophic tyrosine receptor kinase (NTRK) gene fusions. These NTRK gene fusions are oncogenic drivers found in most tumour types at a low frequency (<5%), and at a higher frequency (>80%) in a small number of rare tumours (e.g., secretory carcinoma of the salivary gland and of the breast). They are generally mutually exclusive of other common oncogenic drivers. Larotrectinib and entrectinib have demonstrated impressive overall response rates and tolerability in Phase I/II trials in patients with TRK fusion cancer with no other effective treatment options. Given the low frequency of TRK fusion cancer and the heterogeneous molecular testing landscape in Canada, identifying and optimally managing such patients represents a new challenge. We provide a Canadian consensus on when and how to test for NTRK gene fusions and when to consider treatment with a TRK inhibitor. We focus on five tumour types: thyroid carcinoma, colorectal carcinoma, non-small cell lung carcinoma, soft tissue sarcoma, and salivary gland carcinoma. Based on the probability of the tumour harbouring an NTRK gene fusion, we also suggest a tumour-agnostic consensus for NTRK gene fusion testing and treatment. We recommend considering a TRK inhibitor in all patients with TRK fusion cancer with no other effective treatment options.


A Comparison of Three Computer-based Methods Used to Determine EMG Signal Amplitude.
Electromyography is a commonly used method to determine relative effort and neuromuscular drive to skeletal muscle. A limitation of the interpretation of EMG within the literature is the many methods used to determine the intensity of muscle activation. In the current study, ten healthy young adults performed a level walking task while EMG was recorded from the tibialis anterior, medial gastrocnemius and fibularis longus. The EMG data were rectified and smoothed using the root mean squared (RMS). Peak RMS (pRMS), mean RMS (mRMS) and integrated EMG (iEMG) were normalized to the peak value within the subject and were used to determine EMG amplitude. A 3×3 repeated measures analysis of variance was used to determine significant differences between the methods of determining EMG amplitude. The findings of the current study show that pRMS produced significantly lower EMG amplitudes than mRMS or iEMG values. Furthermore, mRMS and iEMG produced nearly identical normalized EMG amplitudes. Based on the findings of this study and the components of each measurement of EMG amplitude, it is suggested to use mRMS to determine EMG amplitude.


Epidemic of Isothiazolinone Allergy in North America: Prevalence Data From the North American Contact Dermatitis Group, 2013-2014.
Preservative sensitivity patterns evolve with changing use patterns in products. During the last decade, the use of methylisothiazolinone (MI) at higher concentrations in both leave-on and rinse-off products has significantly increased. This is the first North American Contact Dermatitis Group reporting cycle that includes both methylchloroisothiazolinone (MCI)/MI and MI data. The aim of this study was to report the prevalence of isothiazolinone allergy (MCI/MI and MI) in the North American Contact Dermatitis Group patch-test population from January 1, 2013, to December 31, 2014. At 13 centers in North America, 4860 patients were patch tested in a standardized manner with a series of 70 allergens, including MCI/MI 0.01% aqueous (aq) and MI 0.2% aq. Three hundred five patients (6.3%) had a positive reaction to MCI/MI; this is a significant increase from the previous cycle (5.0%, 2011-2012; P = 0.011). Five hundred twenty-one patients (10.7%) had a positive reaction to MI. These 2 isothiazolinones were among the most common preservative allergens in the 2013 to 2014 cycle; 11.9% of patch-tested individuals were allergic to 1 or both isothiazolinones. Individuals with MCI/MI and MI allergy were significantly more likely to have occupationally related skin disease (P < 0.0001) and hand dermatitis (P < 0.0001, P = 0.0474). The epidemic of isothiazolinone sensitivity documented in Europe is now in North America. Patch testing with only MCI/MI 0.01% aq will miss approximately half of isothiazolinone allergy cases, whereas testing with only MI 0.2% aq will miss approximately 10% of isothiazolinone allergy cases.


Volume dependence of the dielectric properties of amorphous SiO2.
Using first principles calculations, the analysis of the dielectric properties of amorphous SiO2 (am-SiO2) was performed. We found that the am-SiO2 properties are volume dependent, and the dependence is mainly induced by the variation of nanoporosity at the atomic scale. In particular, both ionic and electronic contributions to the static dielectric constants are functions of volume with clear trends. Moreover, using the unique parameterization of the dielectric function provided in this work, we predict dielectric functions at imaginary frequencies of different SiO2 polymorphs having similar band gap energies.


The effect of nondialyzable material (NDM) cranberry extract on formation of contact lens biofilm by Staphylococcus epidermidis.
To assess the effects of NDM from cranberries on Staphylococcus epidermidis biofilm formed on soft contact lenses. Soft contact lenses were incubated in Tryptic Soy Broth (TSB) together with S. epidermidis (ATCC35984/RP62A) and various concentrations of NDM, and inspected by scanning electron and confocal microscopy. The TSB was collected after sonification and monitored turbidometrically. NDM at ≥500 μg/mL concentration caused a significant (P < 0.01) reduction of biofilm. Scanning electron microscopy of biofilm in the presence of 500 to 1000 μg/mL NDM confirmed these results. In control lenses, multilayered mushroom-shaped biofilm and complete coverage of the lens surface were seen, whereas after incubation with 500 μg NDM per mL TSB, the biofilm was thinner with smaller protuberances, and exposed lens surface was partially seen. In samples incubated with 1000 μg NDM per mL TSB, the lens surface was clearly seen between sporadic microcolonies. NDM reduces formation of biofilm on soft contact lenses. This has important implications for the prevention of contact lens-related corneal infections caused by S. epidermidis.


Shared epitopes between Mycoplasma pneumoniae major adhesin protein P1 and a 140-kilodalton protein of Mycoplasma genitalium.
Previous serological data have demonstrated cross-reactive antigens between two pathogenic species of mycoplasmas, M. pneumoniae and M. genitalium. Preliminary analysis of sera and monoclonal antibodies (MAbs) to protein antigens of these species showed an immunodominance of adhesin P1 (165 kilodaltons [kDa]) of M. pneumoniae in mice and hamsters and a 140-kDa protein of M. genitalium in mice and experimentally infected chimpanzees. To further characterize these two proteins, we assayed multiple anti-P1 and anti-140-kDa protein MAbs by enzyme-linked immunosorbent assay, immunoblot, and radioimmunoprecipitation techniques. The 140-kDa M. genitalium protein was shown to be surface accessible and insensitive to levels of trypsin which readily degrade protein P1. Peptide mapping was used to identify a unique class of MAbs which bound a cross-reactive molecule common to both the major adhesin protein P1 of M. pneumoniae and the 140-kDa protein of M. genitalium. MAbs generated against both M. pneumoniae and M. genitalium which were reactive with this determinant blocked M. pneumoniae attachment to chicken erythrocytes.


Functional expression, purification and reconstitution of the recombinant phosphate transporter Pho89 of Saccharomyces cerevisiae.
The Saccharomyces cerevisiae high-affinity phosphate transporter Pho89 is a member of the inorganic phosphate (Pi) transporter (PiT) family, and shares significant homology with the type III Na(+)/Pi symporters, hPit1 and hPit2. Currently, detailed biochemical and biophysical analyses of Pho89 to better understand its transport mechanisms are limited, owing to the lack of purified Pho89 in an active form. In the present study, we expressed functional Pho89 in the cell membrane of Pichia pastoris, solubilized it in Triton X-100 and foscholine-12, and purified it by immobilized nickel affinity chromatography combined with size exclusion chromatography. The protein eluted as an oligomer on the gel filtration column, and SDS/PAGE followed by western blotting analysis revealed that the protein appeared as bands of approximately 63, 140 and 520 kDa, corresponding to the monomeric, dimeric and oligomeric masses of the protein, respectively. Proteoliposomes containing purified and reconstituted Pho89 showed Na(+)-dependent Pi transport activity driven by an artificially imposed electrochemical Na(+) gradient. This implies that Pho89 operates as a symporter. Moreover, its activity is sensitive to the Na(+) ionophore monensin. To our knowledge, this study represents the first report on the functional reconstitution of a Pi-coupled PiT family member.


Facial soft tissue changes after orthodontic treatment.
To successfully meet expectations on facial esthetics, it is important to understand normal craniofacial growth and the impact of orthodontic treatment thereon. To date, there have been few studies documenting changes in facial esthetics through photography. The objective of this study was to compare facial soft tissue esthetics before and after orthodontic treatment by means of photographic analysis. The 45 children were divided into 3 groups according to Angle's classification: Groups I, II, and III comprised children with class I, II, and III malocclusion, respectively. Photographs were analyzed with a software. Twenty-one soft tissue landmarks were identified on profile and frontal photographs, ratios and angles were calculated. For group I, there was no difference between pre- and post-treatment facial analysis. For group II, there were significant changes in 5 values. The most significant changes were observed for A-N-B and Al-Me/Ch-Me. For group III, we noted significant changes for 5 values. The most significant change was observed for N-Pn-Pog. There were significant changes in facial soft tissue esthetics after orthodontic treatment for class II and III cases. Changes in A-N-B and nose tip angle (N-Pn-Cm) were observed for class II and class III subjects.


Fragmented QRS as a marker of conduction abnormality and a predictor of prognosis of Brugada syndrome.
Conduction abnormalities serve as a substrate for ventricular fibrillation (VF) in patients with Brugada syndrome (BS). Signal-averaged electrograms can detect late potentials, but the significance of conduction abnormalities within the QRS complex is still unknown. The latter can present as multiple spikes within the QRS complex (fragmented QRS [f-QRS]). We hypothesized that f-QRS could indicate a substrate for VF and might predict a high risk of VF for patients with BS. In study 1, we analyzed the incidence of f-QRS in 115 patients with BS (13 resuscitated from VF, 28 with syncope, and 74 asymptomatic). f-QRS was observed in 43% of patients, more often in the VF group (incidence of f-QRS: VF 85%, syncope 50%, and asymptomatic 34%, P<0.01). SCN5A mutations occurred more often in patients with f-QRS (33%) than in patients without f-QRS (5%). In patients with syncope or VF, only 6% without f-QRS experienced VF during follow-up (43+/-25 months), but 58% of patients with f-QRS had recurrent syncope due to VF (P<0.01). In study 2, to investigate the mechanism of f-QRS, we studied in vitro models of BS in canine right ventricular tissues (n=4) and optically mapped multisite action potentials. In the experimental model of BS, ST elevation resulted from a large phase 1 notch of the action potential in the epicardium, and local epicardial activation delay reproduced f-QRS in the transmural ECG. f-QRS appears to be a marker for the substrate for spontaneous VF in BS and predicts patients at high risk of syncope.


Chronic conditions and the decline in late-life disability.
Using data from the 1997-2004 National Health Interview Survey (NHIS), we examine the role of chronic conditions in recent declines in late-life disability prevalence. Building upon prior studies, we decompose disability declines into changes in the prevalence of chronic conditions and in the risk of disability given a condition. In doing so, we extend Kitigawa's (1955) classical decomposition technique to take advantage of the annual data points in the NHIS. Then we use respondents' reports of conditions causing their disability to repartition these traditional decomposition components. We find a general pattern of increasing prevalence of chronic conditions accompanied by declines in the percentage reporting disability among those with a given condition. We also find declines in heart and circulatory conditions, vision impairments, and possibly arthritis and increases in obesity as reported causes of disability. Based on decomposition analyses, we conclude that heart and circulatory conditions as well as vision limitations played a major role in recent declines in late-life disability prevalence and that arthritis may also be a contributing factor. We discuss these findings in light of improvements in treatments and changes in the environments of older adults.


Mini-open lateral retropleural/retroperitoneal approaches for thoracic and thoracolumbar junction anterior column pathologies.
Advancements in less invasive lateral retropleural/retroperitoneal approaches aim to address the limitation of posterolateral approaches and avoid complications associated with anterior open thoracotomy or thoracoabdominal approaches. Consecutive patients treated with a mini-open lateral approach for thoracic or thoracolumbar anterior column pathologies were analyzed in a retrospective case series including clinical and radiographic outcomes. Special attention is given to operative techniques and surgical nuances. Eleven patients underwent a mini-open lateral retropleural or combined retropleural/retroperitoneal approach for thoracic or thoracolumbar junction lesions. Surgical indications included chronic fracture/deformity (n = 5), acute fracture (n = 2), neoplasm (n = 2), and osteomyelitis (n = 2). The mean length of postoperative hospital stay was 7.2 days (range 2-19 days). All patients ultimately had successful decompression and reconstruction with a mean follow-up of 16.7 months (range 6-29 months). Axial back pain assessed by the visual analog scale improved from a mean score of 8.2 to 2.2. Complications included 1 patient with deep venous thrombosis and pulmonary embolism and 1 with pneumonia. One patient developed increased leg weakness, which subsequently improved. One patient undergoing corpectomy with only lateral plate fixation developed cage subsidence requiring posterior stabilization. Mini-open lateral retropleural and retroperitoneal corpectomies can safely achieve anterior column reconstruction and spinal deformity correction for various thoracic and thoracolumbar vertebral pathologies.


Global transcriptional profiling of longitudinal clinical isolates of Mycobacterium tuberculosis exhibiting rapid accumulation of drug resistance.
The identification of multidrug resistant (MDR), extensively and totally drug resistant Mycobacterium tuberculosis (Mtb), in vulnerable sites such as Mumbai, is a grave threat to the control of tuberculosis. The current study aimed at explaining the rapid expression of MDR in Directly Observed Treatment Short Course (DOTS) compliant patients, represents the first study comparing global transcriptional profiles of 3 pairs of clinical Mtb isolates, collected longitudinally at initiation and completion of DOTS. While the isolates were drug susceptible (DS) at onset and MDR at completion of DOTS, they exhibited identical DNA fingerprints at both points of collection. The whole genome transcriptional analysis was performed using total RNA from H37Rv and 3 locally predominant spoligotypes viz. MANU1, CAS and Beijing, hybridized on MTBv3 (BuG@S) microarray, and yielded 36, 98 and 45 differentially expressed genes respectively. Genes encoding transcription factors (sig, rpoB), cell wall biosynthesis (emb genes), protein synthesis (rpl) and additional central metabolic pathways (ppdK, pknH, pfkB) were found to be down regulated in the MDR isolates as compared to the DS isolate of the same genotype. Up regulation of drug efflux pumps, ABC transporters, trans-membrane proteins and stress response transcriptional factors (whiB) in the MDR isolates was observed. The data indicated that Mtb, without specific mutations in drug target genes may persist in the host due to additional mechanisms like drug efflux pumps and lowered rate of metabolism. Furthermore this population of Mtb, which also showed reduced DNA repair activity, would result in selection and stabilization of spontaneous mutations in drug target genes, causing selection of a MDR strain in the presence of drug pressures. Efflux pump such as drrA may play a significant role in increasing fitness of low level drug resistant cells and assist in survival of Mtb till acquisition of drug resistant mutations with least fitness cost.


[Antimicrobial resistance of the most frequently isolated microorganisms in the Hospital General La Mancha Centro between June 2009 and May 2010].
During the last two decades an increased incidence of infections caused by multiresistant bacteria has been observed. The spread of these microorganisms in the hospital is a major therapeutic and epidemiological problem. The aim of this study was to determine local resistance patterns of microorganisms causative of multirresistant infections in patients admitted to our hospital. A retrospective study was designed, including Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and Acinetobacter baumannii strains isolated from clinical and surveillance samples of patients admitted to the Hospital General La Mancha Centro, between June 2009 and May 2010. The rate of S. aureus isolates resistant to oxacillin was 50%, with 0% resistance to vancomycin. The percentage of resistance to 3rd generation cephalosporins in E. coli and K. pneumoniae was 17 and 19%, respectively; the ESBL-production in enterobacterial strains was 15 and 19%, respectively, and the quinolone resistance was 41 and 28%, respectively. The resistance of P. aeruginosa to ceftazidime and imipenem was 30 and 40%, respectively. Most strains of A. baumannii studied came from a single multidrug-resistant clone, endemic in the ICU of our hospital. Of particular concern is the high rate of MRSA, E. coli and K. pneumoniae ESBL producers and resistant to fluoroquinolones as well as P. aeruginosa multiresistant. A. baumannii isolates belong mainly to endemic multidrug-resistant clone from the ICU.


Autoregulation of neuroblastoma growth by vasoactive intestinal peptide.
Elevated serum levels of vasoactive intestinal peptide (VIP) are associated with some cases of neuroblastoma and correlate with a favorable prognosis. VIP has previously been shown in our laboratory to cause the in vitro growth inhibition and morphological differentiation of the human neuroblastoma cell line, LA-N-5. It is now shown that LA-N-5 cells express immunoreactive VIP and bear specific VIP receptors. Antagonism of endogenous VIP, either by competitive inhibition or receptor blockade, increased cell proliferation, suggesting that VIP is operative in normal growth regulation. Intracellular and extracellular levels of VIP were also shown to increase significantly during the retinoic acid-induced differentiation of these cells. Furthermore, a concomitant marked increase in VIP receptor expression was demonstrated with cellular differentiation. These receptors remain functional as evidenced by a matching increase in the level of detectable cAMP generated in response to exogenous VIP. It is concluded that VIP is a normal autoregulator of neuroblastoma cell growth and differentiation, and that retinoic acid-mediated differentiation may be, in part, due to endogenous VIP.


Age- and gender-specific prevalence of depression in latest-life--systematic review and meta-analysis.
The objective of the study is to systematically analyze the prevalence of depression in latest life (75+), particularly focusing on age- and gender-specific rates across the latest-life age groups. Relevant articles were identified by systematically searching the databases MEDLINE, Web of Science, Cochrane Library and Psycinfo and relevant literature from 1999 onwards was reviewed. Studies based on the community-based elderly population aged 75 years and older were included. Quality of studies was assessed. Meta-analysis was performed using random effects model. 24 studies reporting age- and gender-specific prevalence of depression were found. 13 studies had a high to moderate methodical quality. The prevalence of major depression ranged from 4.6% to 9.3%, and that of depressive disorders from 4.5% to 37.4%. Pooled prevalence was 7.2% (95% CI 4.4-10.6%) for major depression and 17.1% (95% CI 9.7-26.1%) for depressive disorders. Potential sources of high heterogeneity of prevalence were study design, sampling strategy, study quality and applied diagnostics of latest life depression. Despite the wide variation in estimates, it is evident that latest life depression is common. To reduce variability of study results, particularly sampling strategies (inclusion of nursing home residents and severe cognitively impaired individuals) for the old age study populations should be addressed more thoroughly in future research.


Persistently highest risk areas for hantavirus pulmonary syndrome: potential sites for refugia.
Interannual variation in the number of cases of human disease caused by hantaviruses in North America has been hypothesized to reflect environmental changes that influence rodent reservoir populations. This hypothesis postulates that when cases are rare reservoir populations are geographically restricted in patches of suitable habitat. Identifying these sites, which is needed to test the hypothesis, has proven to be a challenge. Satellite imagery of the U.S. Southwest has shown associations among the likelihood of human hantaviral disease and increases in the rodent populations, as well as increased prevalence of Sin Nombre virus (SNV) in rodent populations. In this study we characterize local areas that had environmental signatures that persisted as predicted highest risk sites for human disease through much of the 1990s. These areas represent a small percentage (0.3%) of the region. Exploratory analyses indicate that these areas were not randomly distributed, but were associated with certain landscape characteristics. Characteristics of elevation, slope, aspect, and land cover were associated with persistent high risk. Using multivariate Poisson regression to control for confounding effects, sites with deciduous- or mixed-forest land cover on moderate to steep slopes (>5 degrees) above 2130 m elevation were associated with increasing numbers of years at highest risk. These are candidate locations for refugia. Sites associated with cleared ground or shrubland were less often associated with high risk compared to reference conditions. The seasonal patterns of vegetation growth in persistently high-risk areas were compared to matched locations using MODIS (moderate resolution imaging spectroradiometer) NDVI (normalized difference vegetation index) during a time of a severe drought in the region from 2002 to 2004. Despite the drought and regardless of land cover, the NDVI in persistently highest risk areas had an early onset, with significantly higher levels of green vegetation that lasted longer than at comparable sites. These observations identify locations that can be monitored for the abundance of P. maniculatus and presence of SNV. If these sites are refugia, we predict they will be occupied by infected deer mice when other monitored sites are unoccupied.


Genotypes do not confer risk for delinquency but rather alter susceptibility to positive and negative environmental factors: gene-environmentinteractions of BDNF Val66Met, 5-HTTLPR, and MAOA-uVNTR [corrected].
Previous evidence of gene-by-environment interactions associated with emotional and behavioral disorders is contradictory. Differences in findings may result from variation in valence and dose of the environmental factor, and/or failure to take account of gene-by-gene interactions. The present study investigated interactions between the brain-derived neurotrophic factor gene (BDNF Val66Met), the serotonin transporter gene-linked polymorphic region (5-HTTLPR), the monoamine oxidase A (MAOA-uVNTR) polymorphisms, family conflict, sexual abuse, the quality of the child-parent relationship, and teenage delinquency. In 2006, as part of the Survey of Adolescent Life in Västmanland, Sweden, 1 337 high-school students, aged 17-18 years, anonymously completed questionnaires and provided saliva samples for DNA analyses. Teenage delinquency was associated with two-, three-, and four-way interactions of each of the genotypes and the three environmental factors. Significant four-way interactions were found for BDNF Val66Met × 5-HTTLPR×MAOA-uVNTR × family conflicts and for BDNF Val66Met × 5-HTTLPR×MAOA-uVNTR × sexual abuse. Further, the two genotype combinations that differed the most in expression levels (BDNF Val66Met Val, 5-HTTLPR LL, MAOA-uVNTR LL [girls] and L [boys] vs BDNF Val66Met Val/Met, 5-HTTLPR S/LS, MAOA-uVNTR S/SS/LS) in interaction with family conflict and sexual abuse were associated with the highest delinquency scores. The genetic variants previously shown to confer vulnerability for delinquency (BDNF Val66Met Val/Met × 5-HTTLPR S × MAOA-uVNTR S) were associated with the lowest delinquency scores in interaction with a positive child-parent relationship. Functional variants of the MAOA-uVNTR, 5-HTTLPR, and BDNF Val66Met, either alone or in interaction with each other, may be best conceptualized as modifying sensitivity to environmental factors that confer either risk or protection for teenage delinquency.


MR findings in growth hormone deficiency: correlation with severity of hypopituitarism.
Growth hormone deficiency may present as an isolated deficit (IGHD) or in association with multiple deficiencies (MPHD). Previous studies have not compared the MR imaging findings with the severity of hypopituitarism. Our purpose was to determine whether MR imaging can distinguish between IGHD and MPHD. Forty-four patients with growth hormone deficiency who were examined by MR imaging were included in this retrospective study. On the basis of the endocrinologic findings, 21 were determined to have IGHD and 23 to have MPHD. The presence, size, location, and morphologic characteristics of the stalk, the neurohypophysis, and the adenohypophysis were recorded in each case. Findings in the two groups were compared. Statistical significance was determined by t-test. The stalk was normal in one patient with IGHD and in none of those with MPHD; it was truncated or thin in 19 patients with IGHD (90%) and in only one with MPHD (4%); it was absent in 22 patients with MPHD (96%) and in only one patient with IGHD (5%). These differences between the two groups were highly significant. In 81% of the IGHD patients and in 91% of the MPHD patients the location of the neurohypophysis was ectopic. This difference between the two groups was not significant. Among IGHD patients, the adenohypophysis was of normal size in 13 patients (62%), small in six (29%), and absent in two (9%); the corresponding findings in MPHD patients were seven (30%), six (26%), and 10 (44%). The majority of IGHD patients had a truncated or thin stalk and a normal or small adenohypophysis. An absent stalk and adenohypophysis are characteristic of MPHD. MR imaging can contribute to the prediction of the pattern and severity of hypopituitarism in patients with growth hormone deficiency.


Screening urine samples for the absence of urinary tract infection using the sediMAX automated microscopy analyser.
Urinalysis culminates in a workload skew within the clinical microbiology laboratory. Routine processing involves screening via manual microscopy or biochemical dipstick measurement, followed by culture for each sample. Despite this, as many as 80% of specimens are reported as negative; thus, there is vast wastage of resources and time, as well as delayed turnaround time of results as numerous negative cultures fulfil their required incubation time. Automation provides the potential for streamlining sample screening by efficiently (>30% sample exclusion) and reliably [negative predictive value (NPV) ≥ 95%] ruling out those likely to be negative, whilst also reducing resource usage and hands-on time. The present study explored this idea by using the sediMAX automated microscopy urinalysis platform. We prospectively collected and processed 1411 non-selected samples directly after routine laboratory processing. The results from this study showed multiple optimum cut-off values for microscopy. However, although optimum cut-off values permitted rule-out of 40.1% of specimens, an associated 87.5% NPV was lower than the acceptable limit of 95%. Sensitivity and specificity of leukocytes and bacteria in determining urinary tract infection was assessed by receiver operator characteristic curves with area under the curve values found to be 0.697 [95% confidence interval (CI): 0.665-0.729] and 0.587 (95% CI: 0.551-0.623), respectively. We suggested that the sediMAX was not suitable for use as a rule-out screen prior to culture and further validation work must be carried out before routine use of the analyser.


Comparing the Prognostic Value of the Eighth Edition of the American Joint Committee on Cancer/Tumor Node Metastasis Staging System Between Papillary and Follicular Thyroid Cancer.
Recently, the eighth edition of the American Joint Committee on Cancer (AJCC)/tumor node metastasis (TNM) staging system for differentiated thyroid cancer (DTC) was published. Studies evaluating this new edition have so far only comprised patients with papillary thyroid cancer (PTC) or made no distinction between PTC and follicular thyroid cancer (FTC). Therefore, this study evaluated the prognostic value of the eighth edition of the AJCC/TNM staging system in a European population with DTC, focusing on potential differences between PTC and FTC. Adult patients with DTC who were diagnosed and/or treated at a Dutch university hospital between January 2002 and April 2016 were retrospectively studied. Overall survival (OS) and disease-specific survival (DSS) were analyzed for DTC and for PTC and FTC separately according to the seventh and eighth editions using the Kaplan-Meier method. Cox's proportional hazards model was used to compare the effect of PTC and FTC on survival. The statistical model performance was assessed using the C-index, Akaike information criterion (AIC), and the Bayesian information criterion. The study included 792 patients with DTC (79% PTC, 21% FTC) with mean age of 49 years. Median follow-up was 7.2 years. Reclassification using the eighth edition resulted in the downstaging of 282 (36%) patients, an increased number of patients in stages I and II, and an equivalent decrease in patients with stages III and IV. For DTC, as well as for PTC and FTC separately, stage at diagnosis was significantly related to both OS and DSS (p < 0.001). When using the seventh edition, FTC patients had a significantly lower survival rate than PTC patients in stage I and stage IV for OS, and in stage IV for DSS. This difference in survival rates disappeared using the eighth edition. In general, the statistical model performance was better for the eighth than for the seventh edition. In a European population of patients with DTC, the eighth edition of the AJCC/TNM staging system is a better predictor for both OS and DSS than the previous seventh edition for both PTC and FTC. Furthermore, differences in survival rates between PTC and FTC that were present using the seventh edition disappeared using the eighth edition, implying that this new edition is predicting well, regardless of DTC subtype.


Sorting of Fas ligand to secretory lysosomes is regulated by mono-ubiquitylation and phosphorylation.
Fas ligand (FasL), a potent mediator of apoptosis expressed by CTL and NK cells, is sorted into the inner vesicles of secretory lysosomes for release via exosome-like vesicles. Previous studies identified a proline-rich domain in the cytoplasmic tail required for sorting FasL to secretory lysosomes, but the mechanisms by which this occurs have not been identified. Here we demonstrate that the PRD of FasL binds Fgr, Fyn and Lyn tyrosine kinases, leading to phosphorylation of FasL. Loss of phosphorylation reduces internalisation of FasL into multivesicular bodies. FasL is also directly mono-ubiquitylated at lysines flanking the PRD and mutation of these lysines reduces MVB localisation of FasL. Phosphorylation is not required for ubiquitylation because FasL lacking all tyrosines undergoes mono-ubiquitylation. These studies show that phosphorylation and ubiquitin signals regulate the sorting of FasL to secretory lysosomes by controlling entry into multivesicular bodies.


Glycated hemoglobin and risk of hypertension in the atherosclerosis risk in communities study.
Diabetes and hypertension often co-occur and share risk factors. Hypertension is known to predict diabetes. However, hyperglycemia also may be independently associated with future development of hypertension. We investigated glycated hemoglobin (HbA(1c)) as a predictor of incident hypertension. We conducted a prospective analysis of 9,603 middle-aged participants in the Atherosclerosis Risk in Communities Study without hypertension at baseline. Using Cox proportional hazards models, we estimated the association between HbA(1c) at baseline and incident hypertension by two definitions 1) self-reported hypertension during a maximum of 18 years of follow-up and 2) measured blood pressure or hypertension medication use at clinic visits for a maximum of 9 years of follow-up. We observed 4,800 self-reported and 1,670 visit-based hypertension cases among those without diagnosed diabetes at baseline. Among those with diagnosed diabetes at baseline, we observed 377 self-reported and 119 visit-based hypertension cases. Higher baseline HbA(1c) was associated with an increased risk of hypertension in subjects with and without diabetes. Compared with nondiabetic adults with HbA(1c) <5.7%, HbA(1c) in the prediabetic range (5.7-6.4%) was independently associated with incident self-reported hypertension (hazard ratio 1.14 [95% CI 1.06-1.23]) and visit-detected hypertension (1.17 [1.03-1.33]). We observed that individuals with elevated HbA(1c), even without a prior diabetes diagnosis, are at increased risk of hypertension. HbA(1c) is a known predictor of incident heart disease and stroke. Our results suggest that the association of HbA(1c) with cardiovascular risk may be partially mediated by the development of hypertension.


Catalytic asymmetric desymmetrization approaches to enantioenriched cyclopentanes.
Catalytic asymmetric desymmetrization represents an excellent strategy for accessing highly functionalized chiral building blocks. However, the application of desymmetrization for the synthesis of enantioenriched cyclopentane derivatives remained limited, when compared to chiral cyclohexanes. We have recently developed a desymmetrization protocol for prochiral 2,2-disubstituted cyclopentene-1,3-diones by direct catalytic asymmetric vinylogous nucleophilic addition of deconjugated butenolides. In this perspective, we give an overview of asymmetric desymmetrization reactions leading to enantioenriched cyclopentanes and their derivatives. The focus is kept confined to the diverse nature of reactions used for this purpose. A brief discussion on the potential future directions is also provided.


Endocrino-metabolic features in women with polycystic ovary syndrome during pregnancy.
To elucidate the mechanism of metabolic adaptation of women with polycystic ovary syndrome (PCOS) during pregnancy, the endocrino-metabolic features of a group of PCOS patients with or without gestational diabetes were studied longitudinally during the three trimesters of gestation. Oral glucose tolerance test (OGTT, 100 g) and hyperinsulinaemic-euglycaemic clamp were performed throughout the study. Plasma concentrations of insulin and glucose were determined by radioimmunoassay and glucose oxidase technique, respectively. Five of 13 PCOS patients developed gestational diabetes (GD) at the third trimester (PCOS-GD), while the other eight patients did not develop any alteration of glucose metabolism (PCOS-nGD). Both fasting glucose and insulin plasma concentrations did not change significantly during pregnancy and no difference was seen between the two groups. On the contrary PCOS-GD group early exhibited higher values of area under the curve (AUC) for glucose and insulin response to OGTT with respect to those found in PCOS-nGD group. This difference was already significant in the first gestational trimester. Moreover insulin sensitivity value (M) was significantly lower in the first trimester of gestation in PCOS-GD as compared with that found in PCOS-nGD group. However, as gestation proceeded, M value decreased in PCOS-nDG group and the difference from PCOS patients developing gestational diabetes was not sustained into the second and third trimesters. Both groups had similar body mass index values and AUC insulin increase from first to third trimester of gestation. It is concluded that early alteration of insulin sensitivity and secretion constitute specific risk factors in PCOS patients for the development of abnormalities of glucose tolerance.


Comparative Study to Determine the Prevalence and Socio-Cultural Practices of Infertility in Rural and Urban Field Practice Area of Tertiary Care Hospital, Vijayapura, Karnataka.
Infertility is an iceberg phenomenon where the majority of couples are undiagnosed. The main challenge in estimating the actual burden of infertility is the paucity of population-based studies. Various sociocultural practices such as believing infertility as curse and seeking healing from supernatural powers are still predominant in the community. Hence, the present study was undertaken to know the prevalence and sociocultural practices of infertility in field practice area. A cross-sectional study was conducted in rural and urban field practice areas of tertiary care hospital. A complete enumeration of all the houses was done to list all the eligible couples residing in the area; among them, those at risk of pregnancy were identified so as to find out couples with either primary or secondary infertility. Data were tabulated and analyzed using the SPSS software version 16. The results were expressed in terms of percentages, and Fisher's exact test was used wherever applicable. The prevalence of infertility in rural area was 7.6% and in urban slum, it was 8.8%. Most of them think infertility as a result of past sins and practice unscientific methods to overcome the problem. Infertility stigma is high and there is no proper knowledge about infertility, fertile period and most of them think it as result of past sins.


Characterization of mercury bioremediation by transgenic bacteria expressing metallothionein and polyphosphate kinase.
The use of transgenic bacteria has been proposed as a suitable alternative for mercury remediation. Ideally, mercury would be sequestered by metal-scavenging agents inside transgenic bacteria for subsequent retrieval. So far, this approach has produced limited protection and accumulation. We report here the development of a transgenic system that effectively expresses metallothionein (mt-1) and polyphosphate kinase (ppk) genes in bacteria in order to provide high mercury resistance and accumulation. In this study, bacterial transformation with transcriptional and translational enhanced vectors designed for the expression of metallothionein and polyphosphate kinase provided high transgene transcript levels independent of the gene being expressed. Expression of polyphosphate kinase and metallothionein in transgenic bacteria provided high resistance to mercury, up to 80 μM and 120 μM, respectively. Here we show for the first time that metallothionein can be efficiently expressed in bacteria without being fused to a carrier protein to enhance mercury bioremediation. Cold vapor atomic absorption spectrometry analyzes revealed that the mt-1 transgenic bacteria accumulated up to 100.2 ± 17.6 μM of mercury from media containing 120 μM Hg. The extent of mercury remediation was such that the contaminated media remediated by the mt-1 transgenic bacteria supported the growth of untransformed bacteria. Cell aggregation, precipitation and color changes were visually observed in mt-1 and ppk transgenic bacteria when these cells were grown in high mercury concentrations. The transgenic bacterial system described in this study presents a viable technology for mercury bioremediation from liquid matrices because it provides high mercury resistance and accumulation while inhibiting elemental mercury volatilization. This is the first report that shows that metallothionein expression provides mercury resistance and accumulation in recombinant bacteria. The high accumulation of mercury in the transgenic cells could present the possibility of retrieving the accumulated mercury for further industrial applications.


Structural basis of the selective block of Kv1.2 by maurotoxin from computer simulations.
The 34-residue polypeptide maurotoxin (MTx) isolated from scorpion venoms selectively inhibits the current of the voltage-gated potassium channel Kv1.2 by occluding the ion conduction pathway. Here using molecular dynamics simulation as a docking method, the binding modes of MTx to three closely related channels (Kv1.1, Kv1.2 and Kv1.3) are examined. We show that MTx forms more favorable electrostatic interactions with the outer vestibule of Kv1.2 compared to Kv1.1 and Kv1.3, consistent with the selectivity of MTx for Kv1.2 over Kv1.1 and Kv1.3 observed experimentally. One salt bridge in the bound complex of MTx-Kv1.2 forms and breaks in a simulation period of 20 ns, suggesting the dynamic nature of toxin-channel interactions. The toxin selectivity likely arises from the differences in the shape of the channel outer vestibule, giving rise to distinct orientations of MTx on block. Potential of mean force calculations show that MTx blocks Kv1.1, Kv1.2 and Kv1.3 with an IC(50) value of 6 µM, 0.6 nM and 18 µM, respectively.


Estimating Total Corneal Astigmatism From Anterior Corneal Data.
To determine keratometric astigmatism (KA), posterior corneal astigmatism (PCA), and total corneal astigmatism (TCA) in 951 normal eyes, to establish a model for estimating TCA from anterior corneal data, and to test this method in a new group of eyes with toric intraocular lenses (TIOLs). We used a Pentacam HR (high-resolution) Scheimpflug camera to determine KA, PCA, and TCA in 951 normal eyes. A model to estimate TCA from anterior corneal data was evaluated by the difference (=error) between the measured TCA and the estimated value. The model was tested in 40 eyes with TIOLs. KA, TCA, and PCA averaged 1.06 (±0.85) D, 1.05 (±0.83) D, and 0.33 (±0.17) D. The error of the model to estimate TCA averaged zero with an SD of ±0.21 D. Application of this model and of direct Pentacam TCA measurements in TIOL calculation gave similar results, namely a slight reduction of overcorrection in with-the-rule astigmatism, but an eradication of undercorrection in against-the-rule astigmatism. It was possible to estimate TCA accurately from anterior corneal data with a new formula. However, application of both this model on keratometric data and of direct Pentacam measurements in a group of 40 eyes with TIOLs did not completely eradicate the refractive error in TIOL calculation. More studies comparing Pentacam TCA and refractive astigmatism are required.


Detailed Anatomical Orientations for Certain Types of Morphometric Measurements Can Be Determined Automatically With Geometric Algorithms.
Morphometric datasets only convey useful information about variation when measurement landmarks and relevant anatomical axes are clearly defined. We propose that anatomical axes of 3D digital models of bones can be standardized prior to measurement using an algorithm that automatically finds a universal geometric alignment among sampled bones. As a case study, we use teeth of "prosimian" primates. In this sample, equivalent occlusal planes are determined automatically using the R-package auto3dgm. The area of projection into the occlusal plane for each tooth is the measurement of interest. This area is used in computation of a shape metric called relief index (RFI), the natural log of the square root of crown area divided by the square root of occlusal plane projection area. We compare mean and variance parameters of area and RFI values computed from these automatically orientated tooth models with values computed from manually orientated tooth models. According to our results, the manual and automated approaches yield extremely similar mean and variance parameters. The only differences that plausibly modify interpretations of biological meaning slightly favor the automated treatment because a greater proportion of differences among subsamples in the automated treatment are correlated with dietary differences. We conclude that-at least for dental topographic metrics-automated alignment recovers a variance pattern that has meaning similar to previously published datasets based on manual data collection. Therefore, future applications of dental topography can take advantage of automatic alignment to increase objectivity and repeatability.