Rich micronutrient fortification of locally produced infant food does not improve mental and motor development of Zambian infants: a randomised controlled trial.
It is uncertain whether multiple micronutrients benefit the mental and psychomotor development of young children in developing countries. We conducted a randomised double-blind controlled trial to evaluate the effect of a richly micronutrient-fortified v. a basal fortified porridge on mental and psychomotor development in Zambian infants. Infants (n 743) were randomised at age 6 months to receive either the richly fortified or the basal fortified infant food and were followed up until 18 months of age. All the infants were evaluated monthly for achievement of a series of developmental milestones. The Bayley scales of infant development II were administered to a subsample of 502 infants at 6, 12 and 18 months. Rich micronutrient fortification had no significant benefit on the following: (a) number of developmental milestones achieved (rate ratio at 12 months = 1·00; 95 % CI 0·96, 1·05; P = 0·81, adjusted for sex, socio-economic status and maternal education, with similar results at 15 and 18 months); (b) ages of walking unsupported (hazard ratio (HR) 1·04; 95 % CI 0·88, 1·24; P = 0·63, adjusted for the above covariates) and of speaking three or four clear words (HR 1·01; 95 % CI 0·84, 1·20; P = 0·94, adjusted for the above covariates); (c) mental development index (MDI) and psychomotor development index (PDI) of the Bayley scales (scores difference adjusted for baseline scores, age at the assessment, sex, socio-economic status, maternal education, language, age and HIV status: MDI 0·3 (95 % CI - 0·5, 1·1), P = 0·43; PDI - 0·1 (95 % CI - 0·9, 0·7), P = 0·78). In conclusion, the results do not support the hypothesis that rich micronutrient fortification improves Zambian infants' mental and motor development.


A facile protocol for the immobilisation of vesicles, virus particles, bacteria, and yeast cells.
Immobilisation of liposomes and cells is often a prerequisite for long-term observations. The most common immobilisation approaches rely on surface modifications, encapsulation in porous materials or trapping in microfluidic channels by means of hurdle-like structures. While these approaches are useful for larger mammalian cells, the immobilisation of smaller organisms like bacteria and yeast or membrane model systems such as liposomes typically requires modification of their outer membrane to ensure that they are stably arrested at a defined position. Here, we present a protocol to immobilise biological objects, which can interact with hydrophobic cholesterol. A water-soluble molecule (cholesterol-PEG-biotin) is used as a linker, which can bind via avidin to biotinylated BSA (bBSA) previously absorbed on a glass surface. For better visualization, bBSA is arranged in a dot pattern by means of microcontact printing, and a microfluidic channel is used for sample supply. We show that our approach can be used to successfully immobilise artificial liposomes of different sizes, native (cell-derived) vesicles, vaccinia virions, Saccharomyces cerevisiae and Escherichia coli, simply by flushing the objects through the channel. Under these conditions, small liposomes and biological objects are stably arrested at high flow rates, while larger cells and liposomes can be released again by application of high shear stress. This protocol can be applied for long-term studies where fluids must be changed repeatedly, for measuring fast kinetics where rapid fluid exchange is essential, and to study the effects of shear stress.


Isolation and characterization of glycolipids from some photosynthetic bacteria.
Glycosyl glycerides have been found in substantial amounts in Chloropseudomonas ethylicum but could not be detected in two strains of Rhodopseudomonas palustris. Rhodospirillum molischianum possibly contains small amounts of monoglycosyl diglyceride. The glycolipids of C. ethylicum have been separated into two components. One of these, glycolipid I, is a monogalactosyl diglyceride. Glycolipid II, upon acid hydrolysis, yields galactose, rhamnose, and a third, unidentified sugar. The glycolipids or total lipids of the photosynthetic bacteria examined contained saturated and monounsaturated, but none of the more highly unsaturated, fatty acids.


Bariatric surgery versus intensive medical therapy in obese patients with diabetes.
Observational studies have shown improvement in patients with type 2 diabetes mellitus after bariatric surgery. In this randomized, nonblinded, single-center trial, we evaluated the efficacy of intensive medical therapy alone versus medical therapy plus Roux-en-Y gastric bypass or sleeve gastrectomy in 150 obese patients with uncontrolled type 2 diabetes. The mean (±SD) age of the patients was 49±8 years, and 66% were women. The average glycated hemoglobin level was 9.2±1.5%. The primary end point was the proportion of patients with a glycated hemoglobin level of 6.0% or less 12 months after treatment. Of the 150 patients, 93% completed 12 months of follow-up. The proportion of patients with the primary end point was 12% (5 of 41 patients) in the medical-therapy group versus 42% (21 of 50 patients) in the gastric-bypass group (P=0.002) and 37% (18 of 49 patients) in the sleeve-gastrectomy group (P=0.008). Glycemic control improved in all three groups, with a mean glycated hemoglobin level of 7.5±1.8% in the medical-therapy group, 6.4±0.9% in the gastric-bypass group (P<0.001), and 6.6±1.0% in the sleeve-gastrectomy group (P=0.003). Weight loss was greater in the gastric-bypass group and sleeve-gastrectomy group (-29.4±9.0 kg and -25.1±8.5 kg, respectively) than in the medical-therapy group (-5.4±8.0 kg) (P<0.001 for both comparisons). The use of drugs to lower glucose, lipid, and blood-pressure levels decreased significantly after both surgical procedures but increased in patients receiving medical therapy only. The index for homeostasis model assessment of insulin resistance (HOMA-IR) improved significantly after bariatric surgery. Four patients underwent reoperation. There were no deaths or life-threatening complications. In obese patients with uncontrolled type 2 diabetes, 12 months of medical therapy plus bariatric surgery achieved glycemic control in significantly more patients than medical therapy alone. Further study will be necessary to assess the durability of these results. (Funded by Ethicon Endo-Surgery and others; ClinicalTrials.gov number, NCT00432809.).


Requirement of FoxD3-class signaling for neural crest determination in Xenopus.
Fox factors (winged-helix transcription factors) play important roles in early embryonic patterning. We show here that FoxD3 (Forkhead 6) regulates neural crest determination in Xenopus embryos. Expression of FoxD3 in the presumptive neural crest region starts at the late gastrula stage in a manner similar to that of Slug, and overlaps with that of Zic-r1. When overexpressed in the embryo and in ectodermal explants, FoxD3 induces expression of neural crest markers. Attenuation of FoxD3-related signaling by a dominant-negative FoxD3 construct (FoxD3delN) inhibits neural crest differentiation in vivo without suppressing the CNS marker Sox2. Interestingly, these loss-of-function phenotypes are reversed by coinjecting SLUG: In animal cap explants, neural crest differentiation induced by Slug and Wnt3a is also inhibited by FoxD3delN but not by a dominant-negative form of XBF2. Loss-of-function studies using dominant-negative forms of FoxD3 and Slug indicate that Slug induction by Zic factors requires FoxD3-related signaling, and that FoxD3 and Slug have different requirements in inducing downstream neural crest markers. These data demonstrate that FoxD3 (or its closely related factor) is an essential upstream regulator of neural crest determination.


Factors associated with sexual health and well being in older adulthood.
To provide an update of recent studies on factors associated with sexual well being in older people with a special focus on sexual activity, satisfaction and function. Most recent studies confirmed the relationship between mental health status, especially negative affect and depressive symptoms, and sexual health in older adulthood. However, when this relationship is investigated more deeply, it seems that in fact positive psychological well being (positive affect and quality of life) accounts for sexual activity rather than the lack of depressive symptoms. Moreover, recent studies provided more insight into the relationship between marital characteristics, religion, cognitive functioning and sleeping difficulties and different dimensions of sexual health in older adulthood. In summary, there is substantial previous research revealing associations between various psychosocial, health-related and demographic variables and sexual health in older adulthood. Most considered variables are, for example, age, sex, general physical and mental health. For future research, it is important to consider that relationships between specific variables and sexual health in higher age are usually more complex than they are expected to be and factors differ between different dimensions of sexual health. Communication about sexuality between health-care providers and older patients still implies a lot of barriers and lack of knowledge. Therefore, the provision of communication training for health-care providers to older people in which knowledge is gained about correlates of sexual health in older adulthood should be implemented.


Lexically guided phonetic retuning of foreign-accented speech and its generalization.
Listeners use lexical knowledge to retune phoneme categories. When hearing an ambiguous sound between /s/ and /f/ in lexically unambiguous contexts such as gira[s/f], listeners learn to interpret the sound as /f/ because gira[f] is a real word and gira[s] is not. Later, they apply this learning even in lexically ambiguous contexts (perceiving knife rather than nice). Although such retuning could help listeners adapt to foreign-accented speech, research has focused on single phonetic contrasts artificially manipulated to create ambiguous sounds; however, accented speech varies along many dimensions. It is therefore unclear whether analogies to adaptation to accented speech are warranted. In the present studies, the to-be-adapted ambiguous sound was embedded in a global foreign accent. In addition, conditions of cross-speaker generalization were tested with focus on the extent to which perceptual similarity between 2 speakers' fricatives is a condition for generalization to occur. Results showed that listeners retune phoneme categories manipulated within the context of a global foreign accent, and that they generalize this short-term learning to the perception of phonemes from previously unheard speakers. However, generalization was observed only when exposure and test speakers' fricatives were sampled across a similar perceptual space.


Introducing supported self-management for depression to primary care in Vietnam: A feasibility study in preparation for a randomized controlled trial.
Although depression is a major contributor to the global burden of disease, services remain scarce in many low- and middle-income countries. In Vietnam, depression services are limited, and the government has recently prioritized primary care and community-based service integration. We conducted a pilot study in 2 districts of Hanoi to test the feasibility of (a) introducing a supported self-management (SSM) intervention for adult depression in primary care in Vietnam, and (b) conducting a randomized controlled trial (RCT) to test the effectiveness of the intervention. We conducted focus groups with providers (n = 16) and community members (n = 32) to assess the appropriateness of an Antidepressant Skills Workbook for use in Vietnam. We trained providers (n = 23) to screen patients using the Self-Reporting Questionnaire-20 (SRQ-20) depression scale and to deliver SSM for a 2-month period. A total of 71 patients were eligible to participate in the study, with depression (SRQ-20) and disability (World Health Organization Disability Assessment Schedule 2.0) scores assessed at baseline and 1 and 2 months. Study results demonstrate the feasibility of conducting a full RCT in Vietnam and suggest that SSM is an appropriate care model for the Vietnamese context. There was a statistically significant decrease in depression symptoms on the SRQ-20 and in functional disability in all domains for the World Health Organization Disability Assessment Schedule 2.). Feasibility study results suggested that a full RCT was warranted. An unanticipated outcome of the study was the uptake of the model by the Ministry of Labor, Invalids, and Social Affairs in 2 additional provinces. (PsycINFO Database Record


Novel Simulation Model That Realizes Arterial and Venous Blood Flow for Ultrasound-Guided Central Venous Catheter Insertion in Children.
We developed and validated a realistic simulation model for ultrasound-guided central venous catheter insertion in children that is easy to build and inexpensive and can automatically reproduce arterial and venous blood flow. The simulation model was constructed with a chicken breast, two DWP-385 water pumps, two types of tubes and a controller. An elastic rubber tourniquet and a silicone tube were connected to each water pump, which generated different continuous flows mimicking those of the pediatric internal carotid artery and internal jugular vein, respectively. Both tubes were inserted into a piece of chicken breast and connected to the controller. Then, we provided a simulation program of ultrasound-guided central venous catheter insertion using our novel model to resident emergency medicine physicians. We also collected data on their knowledge and confidence levels regarding the procedure before and after the simulation via questionnaires utilizing a 5-point Likert scale. The flow patterns of the artery and vein were well demonstrated with our model. A total of 11 resident physicians were enrolled. The knowledge and confidence regarding the discrimination of arteries and veins were significantly improved after training with our simulation model (p-value < 0.01). The subjective similarity and usefulness of our model also scored high on the questionnaire (median: 4; interquartile range in both categories: 4-5). Our novel simulation model is useful and realistic for ultrasound-guided central venous catheter insertion training. Clinical impact: This controlled motor system can be applied to many simulation models of artery and vein circulation.


The role of visual experience in the development of columns in cat visual cortex.
The role of experience in the development of the cerebral cortex has long been controversial. Patterned visual experience in the cat begins when the eyes open about a week after birth. Cortical maps for orientation and ocular dominance in the primary visual cortex of cats were found to be present by 2 weeks. Early pattern vision appeared unimportant because these cortical maps developed identically until nearly 3 weeks of age, whether or not the eyes were open. The naïve maps were powerfully dominated by the contralateral eye, and experience was needed for responses to the other eye to become strong, a process unlikely to be strictly Hebbian. With continued visual deprivation, responses to both eyes deteriorated, with a time course parallel to the well-known critical period of cortical plasticity. The basic structure of cortical maps is therefore innate, but experience is essential for specific features of these maps, as well as for maintaining the responsiveness and selectivity of cortical neurons.


Lipopolysaccharide Exposure Induces Maternal Hypozincemia, and Prenatal Zinc Treatment Prevents Autistic-Like Behaviors and Disturbances in the Striatal Dopaminergic and mTOR Systems of Offspring.
Autism is characterized by social deficits, repetitive behaviors, and cognitive inflexibility. The risk factors appear to include genetic and environmental conditions, such as prenatal infections and maternal dietary factors. Previous investigations by our group have demonstrated that prenatal exposure to lipopolysaccharide (LPS), which mimics infection by gram-negative bacteria, induces autistic-like behaviors. To understand the causes of autistic-like behaviors, we evaluated maternal serum metal concentrations, which are involved in intrauterine development and infection/inflammation. We identified reduced maternal levels of zinc, magnesium, selenium and manganese after LPS exposure. Because LPS induced maternal hypozincemia, we treated dams with zinc in an attempt to prevent or ease the impairments in the offspring. We evaluated the social and cognitive autistic-like behaviors and brain tissues of the offspring to identify the central mechanism that triggers the development of autism. Prenatal LPS exposure impaired play behaviors and T-maze spontaneous alternations, i.e., it induced autistic-like behaviors. Prenatal LPS also decreased tyrosine hydroxylase levels and increased the levels of mammalian target of rapamycin (mTOR) in the striatum. Thus, striatal dopaminergic impairments may be related to autism. Moreover, excessive signaling through the mTOR pathway has been considered a biomarker of autism, corroborating our rat model of autism. Prenatal zinc treatment prevented these autistic-like behaviors and striatal dopaminergic and mTOR disturbances in the offspring induced by LPS exposure. The present findings revealed a possible relation between maternal hypozincemia during gestation and the onset of autism. Furthermore, prenatal zinc administration appears to have a beneficial effect on the prevention of autism.


Vascular endothelial growth factor is an essential molecule for mouse kidney development: glomerulogenesis and nephrogenesis.
Homeostasis of body fluid is maintained by the kidneys, which contain two million glomeruli for blood filtration. A glomerulus is formed by growth of Bowman's capsule harmonized with a capillary during kidney development. The vascular endothelial growth factor (VEGF) is an essential angiogenic cytokine, and VEGF deficiency is known to be fatal in mice in early embryonic stages. As secretions of VEGF from cultured kidneys vary according to developmental stages, the role of VEGF in kidney development was studied in vivo by blocking the endogenous VEGF activity with antibody in newborn mice, in which most organs are already developed but kidneys are still developing. The antibody-treated animals showed normal growth but systemic edema. Vessel formation in the superficial renal cortex was disturbed, nephrogenic areas were diminished, and the number of developing nephrons decreased significantly. Many abnormal glomeruli, lacking capillary tufts, were observed in the antibody-treated mice, and VEGF expression in their Bowman's capsule showed a compensatory increase. These results suggest that VEGF mediates communication between the Bowman's capsule and capillary endothelial cells for developing a glomerulus as well as promoting nephrogenesis. In conclusion, VEGF is likely to be an essential molecule for kidney development, and especially for glomerulogenesis.


Changes in white matter in long-term survivors of severe non-missile traumatic brain injury: a computational analysis of magnetic resonance images.
The aim of this study was to investigate long-term consequences of severe non-missile traumatic brain injury (nmTBI) in patients without macroscopic focal brain lesions (>1.6 cm(3)) on regional white-matter density (WMd), and possible correlations with days of coma and memory performances. T1-weighted magnetic-resonance images (MRI) were acquired in 19 nmTBI patients, 3-113 months following the injury, and in 19 control subjects matched for age and gender. In addition, nmTBI patients underwent a battery of standardised memory tests. The MRIs were processed in a fully automatic system using voxel-by-voxel methods. Corpus callosum, fornix, anterior limb of the internal capsule, superior frontal gyrus, para-hippocampal gyrus, optic radiation and chiasma showed significant WMd reduction in nmTBI when compared to control subjects. None of the correlations between days of coma and memory performance scores with nmTBI voxels value that showed WMd reduction reached significance, with the exception of a significant negative correlation between WMd in the mid body of corpus callosum and short-story delayed recall. We detected reductions in WM density in several brain locations similar to those described in previous post mortem investigations. In addition, we observed WMd reduction in the optic chiasma and in the optic radiations; this finding may reflect transneural degeneration along the visual pathway. The weak correlations between specific anatomical sites of the reduced WMd and behavior may reflect the diffuse nature of the brain damage and/or the different time of onset between behavioral manifestations and neuropathological modifications occurring in nmTBI.


Impact of donor-specific HLA antibodies in transplantation, a review of the literature published in the last three years.
This chapter summarizes some of the recent findings published on the role in organ transplantation of HLA antibodies, and--more important--donor-specific HLA antibodies. The negative impact of both, preformed and de novo DSA is now better recognized in recipients of kidney, heart, lung, liver, pancreas, islet cells and bone marrow transplants. An appropriate design of a schedule to monitor HLA antibodies may identify patients at higher risk for immunological events earlier and allow interventions to avoid later graft loss. The value of strategies like preemptive treatment of antibodies and the use of new agents like bortezomib and eculizumab are of interest and need further investigation.


Antiplaque, Antifungal Effectiveness of Aloevera Among Intellectually Disabled Adolescents: Pilot Study.
Various candida species have been associated with poor oral hygiene and active carious lesions. The purpose of this study was to evaluate the effectiveness of aloe vera compared to triclosan toothpaste against total candida, C. albicans, C. tropicalis, Candida krusei, and plaque/gingivitis among intellectually disabled adolescents over 30 days. A double-blind prospective randomized trial was conducted among 40 intellectually disabled adolescents randomly allocated into aloe vera/triclosan groups. The gingival (Löe and Silness index), plaque (Silness and Löe index), and candidal carriage counts were assessed at baseline and follow-up. Caregivers brushed the participant's teeth twice a day using a modified bass method and refrained from any other oral hygiene practices for at least two hours prior to assessment. Aloe vera-contaiing toothpaste caused significant reductions in gingival inflammation and plaque index scores compared to the triclosan group at the end of 30 days. Also, total candidal counts and C. albicans counts were significantly lower in the aloe vera group compared to triclosan at the end of the 30-day follow-up (P<0.05). Aloe vera-containing toothpaste was effective in reducing plaque, gingivitis, and overall candidal counts compared to triclosan. Moreover, individuals treated with aloe vera experienced improved oral health status without any negative side effects.


A multi-subunit chlamydial vaccine induces antibody and cell-mediated immunity in immunized koalas (Phascolarctos cinereus): comparison of three different adjuvants.
Chlamydial infections represent a major threat to the survival of the koala. Infections caused by Chlamydia pecorum cause blindness, infertility, pneumonia and urinary tract infections and represent a threat to the survival of the species. Little is known about the immune response in koalas, or the safety of commonly used adjuvants for induction of protective systemic and mucosal immunity. of study In the present study, we immunized 18 healthy female koalas subcutaneously with a combination of three chlamydial antigens [major outer membrane protein (MOMP), NrdB and TC0512 (Omp85)] mixed with one of three different adjuvants [Alhydrogel, Immunostimulating Complex (ISC) and TiterMax Gold]. All adjuvants induced strong neutralizing IgG responses in plasma against the three antigens with prolonged responses lasting more than 270 days seen in Alhydrogel and ISC immunized animals. Cloacal IgG responses lasting >270 days were also induced in ISC-immunized animals. Chlamydia-specific peripheral blood mononuclear cell proliferative responses were elicited by both Alhydrogel and ISC, and these lasted >270 days in the ISC group. The data show that a multi-subunit chlamydial vaccine, given subcutaneously, can elicit Chlamydia-specific cell-mediated and antibody responses in the koala demonstrating that the development of a protective vaccine is feasible.


Staphylococcus aureus Virulence Affected by an Alternative Nisin A Resistance Mechanism.
Nisin A is a bacteriocin produced by Lactococcus lactis and is widely used as a food preservative. Staphylococcus aureus has the BraRS-VraDE system that provides resistance against low concentrations of nisin A. BraRS is a two-component system that induces the expression of the ABC transporter VraDE. Previously, we isolated a highly nisin A-resistant strain with increased VraDE expression due to a mutation in braRS In this study, we isolated S. aureus MW2 mutants with BraRS-VraDE-independent nisin A resistance. These mutants, designated SAN2 ( S.aureusnisin resistant) and SAN469, had a mutation in pmtR, which encodes a transcriptional regulator responsible for the expression of the pmtABCD operon. As a result, these mutants exhibited increased expression of PmtABCD, a transporter responsible for the export of phenol-soluble modulin (PSM). Characterization of the mutants revealed that they have decreased susceptibility to human β-defensin-3 (hBD3) and LL37, which are innate immune factors. Additionally, these mutants showed higher hemolytic activity than the original MW2 strain. Furthermore, in a mouse bacteremia model, the SAN2 strain exhibited a lower survival rate than the original MW2 strain. These results indicate that the increased expression of pmtABCD due to a pmtR mutation is an alternative nisin A resistance mechanism that also affects virulence in S. aureusIMPORTANCE Recently, the emergence of antibiotic-resistant bacteria has resulted in serious problems for chemotherapy. In addition, many antibacterial agents, such as disinfectants and food additives, are widely used. Therefore, there is a possibility that bacteria are becoming resistant to some antibacterial agents. In this study, we investigated whether Staphylococcus aureus can become resistant to nisin A, one of the bacteriocins applied as a food additive. We isolated a highly nisin A-resistant strain designated SAN2 that displayed increased expression of Pmt proteins, which are involved in the secretion of virulence factors called phenol-soluble modulins (PSMs). This strain also showed decreased susceptibility to human antimicrobial peptides and increased hemolytic activity. In addition, SAN2 showed increased lethal activity in a mouse bacteremia model. Our study provides new insights into the possibility that the acquisition of resistance against food preservatives may modulate virulence in S. aureus, suggesting that we need to pay more attention to the use of food preservatives together with antibiotics.


The nuclear localization of ERK2 occurs by mechanisms both independent of and dependent on energy.
The mitogen-activated protein (MAP) kinases ERK1 and ERK2 often accumulate in the nuclei of stimulated cells to mediate changes in transcription. The mechanisms underlying stimulus-dependent redistribution of these kinases remain unclear. We have used a permeabilized cell reconstitution assay in HeLa cells and human foreskin fibroblasts to explore the processes by which ERK2 enters and exits the nucleus. We previously reported that entry of unphosphorylated ERK2 into the nucleus occurs by facilitated diffusion not requiring cytosolic transport factors. We find that export, like import, can occur by an energy- and carrier-independent mechanism. An energy-dependent mechanism of ERK2 export can also be distinguished, mediated at least in part through the exportin CRM1. We have also examined import and export of thiophosphorylated, active ERK2. Import of active ERK2 is significantly enhanced by the addition of exogenous transport factors and an energy regeneration system. These studies support a model in which multiple constitutive and regulated processes control the subcellular distribution of ERK2.


Demystifying computer science for molecular ecologists.
In this age of data-driven science and high-throughput biology, computational thinking is becoming an increasingly important skill for tackling both new and long-standing biological questions. However, despite its obvious importance and conspicuous integration into many areas of biology, computer science is still viewed as an obscure field that has, thus far, permeated into only a few of the biology curricula across the nation. A national survey has shown that lack of computational literacy in environmental sciences is the norm rather than the exception [Valle & Berdanier (2012) Bulletin of the Ecological Society of America, 93, 373-389]. In this article, we seek to introduce a few important concepts in computer science with the aim of providing a context-specific introduction aimed at research biologists. Our goal was to help biologists understand some of the most important mainstream computational concepts to better appreciate bioinformatics methods and trade-offs that are not obvious to the uninitiated.


Evaluation of the Wii Balance Board for walking aids prediction: proof-of-concept study in total knee arthroplasty.
To provide proof-of-concept for the validity of the Wii Balance Board (WBB) measures to predict the type of walking aids required by inpatients with a recent (≤4 days) total knee arthroplasty (TKA). A cross-sectional sample of 89 inpatients (mean age, 67.0±8 years) with TKA was analyzed. A multivariable proportional odds prediction model was constructed using 8 pre-specified predictors – namely, age, sex, body mass index, knee pain, knee range-of-motion, active knee lag, and WBB-derived standing balance. The type of walking aids prescribed on day 4 post-surgery was the outcome of interest – an ordinal variable with 4 categories (walking stick, narrow- and broad-base quadstick, and walking frame). Women, increasing body mass index, and poorer standing balance were independently associated with greater odds for requiring walking aids with a larger base-of-support. The concordance-index of the prediction model was 0.74. The model comprising only WBB-derived standing balance had nearly half (44%) the explanatory power of the full model. Adding WBB-derived standing balance to conventional demographic and knee variables resulted in a continuous net reclassification index of 0.60 (95%CI,0.19-1.01), predominantly due to better identification of patients who required walking aids with a large base-of-support (sensitivity gain). The WBB was able to provide quantitative measures of standing balance which could assist healthcare professionals in prescribing the appropriate type of walking aids for patients. Further investigation is needed to assess whether using the WBB could lead to meaningful changes in clinical outcomes such as falls.


Energy content of stools in normal healthy controls and patients with cystic fibrosis.
Stool energy losses and the sources of energy within the stool were determined in 20 healthy controls and 20 patients with cystic fibrosis while on their habitual pancreatic enzyme replacement treatment. Stool energy losses were equivalent to 3.5% of gross energy intake in healthy children (range 1.3-5.8%). Despite a comparable gross energy intake, stool energy losses were three times greater in patients with cystic fibrosis than controls averaging 10.6% of gross energy intake (range 4.9-19.7%). Stool lipid could account for only 29% and 41% of the energy within the stool in controls and patients with cystic fibrosis respectively and was poorly related to stool energy. Approximately 30% of the energy within the stool could be attributable to colonic bacteria in both the healthy children and patients with cystic fibrosis. These results suggest that stool energy losses in healthy children are relatively modest but that even when patients with cystic fibrosis are symptomatically well controlled on pancreatic enzyme replacement, raised stool energy losses may continue to contribute towards an energy deficit sufficient to limit growth in cystic fibrosis. As the energy content per gram wet weight remains relatively constant (8 kJ/g), stool energy losses may be estimated from simple measurements of stool wet weight.


Performance evaluation of two-dimensional phase unwrapping algorithms.
We present a performance evaluation of eight two-dimensional phase unwrapping methods with respect to correct phase unwrapping and execution times. The evaluated methods are block least squares (BLS), adaptive integration (AI), quality guided path following (QUAL), mask cut (MCUT), multigrid (MGRID), preconditioned conjugate gradient (PCG), Flynn's (FLYNN), and Liang's (LIANG). This set included integration- (path following), least-squares-, L(1)-, and model-based methods. The methods were tested on several synthetic images, on two magnetic resonance images, and on two interferometry images. The synthetic images were designed to demonstrate different aspects of the phase unwrapping problem. To test the noise robustness of the methods, independent noise was added to the synthetic images to yield different signal-to-noise ratios. Each experiment was performed 50 times with different noise realizations to test the stability of the methods. The results of the experiments showed that the congruent minimum L(1) norm FLYNN method was best overall and the most noise robust of the methods, but it was also one of the slowest methods. The integration-based QUAL method was the only method that correctly unwrapped the two interferometry images. The least-squares-based methods (MGRID, PCG) gave worse results on average than did the integration- (or path following) based methods (BLS, AI, QUAL, MCUT) and were also slower. The model-based LIANG method was sensitive to noise and resulted in large errors for the magnetic resonance images and the interferometry images. In conclusion, for a particular application there is a trade-off between the quality of the unwrapping and the execution time when we attempt to select the most appropriate method.


Assessing the generalizability of randomized trial results to target populations.
Recent years have seen increasing interest in and attention to evidence-based practices, where the "evidence" generally comes from well-conducted randomized trials. However, while those trials yield accurate estimates of the effect of the intervention for the participants in the trial (known as "internal validity"), they do not always yield relevant information about the effects in a particular target population (known as "external validity"). This may be due to a lack of specification of a target population when designing the trial, difficulties recruiting a sample that is representative of a prespecified target population, or to interest in considering a target population somewhat different from the population directly targeted by the trial. This paper first provides an overview of existing design and analysis methods for assessing and enhancing the ability of a randomized trial to estimate treatment effects in a target population. It then provides a case study using one particular method, which weights the subjects in a randomized trial to match the population on a set of observed characteristics. The case study uses data from a randomized trial of school-wide positive behavioral interventions and supports (PBIS); our interest is in generalizing the results to the state of Maryland. In the case of PBIS, after weighting, estimated effects in the target population were similar to those observed in the randomized trial. The paper illustrates that statistical methods can be used to assess and enhance the external validity of randomized trials, making the results more applicable to policy and clinical questions. However, there are also many open research questions; future research should focus on questions of treatment effect heterogeneity and further developing these methods for enhancing external validity. Researchers should think carefully about the external validity of randomized trials and be cautious about extrapolating results to specific populations unless they are confident of the similarity between the trial sample and that target population.


Functional genomics of chicken, mouse, and human titin supports splice diversity as an important mechanism for regulating biomechanics of striated muscle.
Titin is a giant filamentous elastic protein that spans from the Z-disk to M-band regions of the sarcomere. The I-band region of titin is extensible and develops passive force in stretched sarcomeres. This force has been implicated as a factor involved in regulating cardiac contraction. To better understand the adaptation in the extensible region of titin, we report the sequence and annotation of the chicken and mouse titin genes and compare them to the human titin gene. Our results reveal a high degree of conservation within the genomic region encoding the A-band segment of titin, consistent with the structural similarity of vertebrate A-bands. In contrast, the genomic region encoding the Z-disk and I-band segments is highly divergent. This is most prominent within the central I-band segment, where chicken titin has fewer but larger PEVK exons (up to 1,992 bp). Furthermore, in mouse titin we found two LINE repeats that are inserted in the Z-disk and I-band regions, the regions that account for most of the splice isoform diversity. Transcript studies show that a group of 55 I-band exons is differentially expressed in chicken titin. Consistent with a large degree of titin isoform plasticity and variation in PEVK content, chicken skeletal titins range in size from approximately 3,000 to approximately 3,700 kDa and vary greatly in passive mechanical properties. Low-angle X-ray diffraction experiments reveal significant differences in myofilament lattice spacing that correlate with titin isoform expression. We conclude that titin splice diversity regulates structure and biomechanics of the sarcomere.


Simultaneous pollutant removal and electricity generation in denitrifying microbial fuel cell with boric acid-borate buffer solution.
A double-chamber denitrifying microbial fuel cell (MFC), using boric acid-borate buffer solution as an alternative to phosphate buffer solution, was set up to investigate the influence of buffer solution concentration, temperature and external resistance on electricity generation and pollutant removal efficiency. The result revealed that the denitrifying MFC with boric acid-borate buffer solution was successfully started up in 51 days, with a stable cell voltage of 205.1 ± 1.96 mV at an external resistance of 50 Ω. Higher concentration of buffer solution favored nitrogen removal and electricity generation. The maximum power density of 8.27 W/m(3) net cathodic chamber was obtained at a buffer solution concentration of 100 mmol/L. An increase in temperature benefitted electricity generation and nitrogen removal. A suitable temperature for this denitrifying MFC was suggested to be 25 °C. Decreasing the external resistance favored nitrogen removal and organic matter consumption by exoelectrogens.


The physiological and pharmacological role of presynaptic alpha- and beta-adrenoceptors in man.
Two studies were performed each in six normal volunteers in order to find evidence of either a physiological or pharmacological role of presynaptic alpha- and presynaptic beta-adrenoceptors in man. In Study 1 subjects received a 60 min infusion of guanfacine 3 mg (alpha 2-adrenoceptor agonist) preceded by either idazoxan (alpha 2-adrenoceptor antagonist) or vehicle. Guanfacine reduced plasma noradrenaline concentration by approximately 30% and this fall was not antagonised by the alpha 2-receptor antagonist. The 30-fold increase in plasma growth hormone, measured as a marker of the central action of guanfacine, was almost completely blocked by idazoxan. A comparison of the drug concentrations of idazoxan and guanfacine, together with their relative affinities for alpha 2-adrenoceptors, suggested that the idazoxan could not block the peripheral actions of guanfacine and that these were responsible for the fall in plasma noradrenaline concentration. In Study 2 adrenaline 0.05 micrograms kg-1 min-1 was infused for 80 min preceded by either idazoxan or vehicle. After vehicle, adrenaline caused no change in plasma noradrenaline concentration whereas it rose approximately 25% after administration of idazoxan. This was probably due to unmasking of presynaptic beta-adrenoceptor stimulation by adrenaline when the opposing inhibitory autoreceptor was blocked.


A review and critical analysis of the scientific literature related to 100% fruit juice and human health.
The association between the consumption of pure (100%) fruit juice (PFJ) and human health is uncertain. The current review summarizes data published between 1995 and 2012 related to PFJ with a focus on juices that are widely available and studied in forms representing native juice without supplemental nutrients or enhanced phytochemical content. The effects of apple, cranberry, grape, grapefruit, orange, and pomegranate PFJ intake on outcomes linked to cancer, cardiovascular disease, cognition, hypertension, inflammation, oxidation, platelet function, urinary tract infection, and vascular reactivity are reviewed. Implications for bodyweight regulation are also addressed. The collective data are provocative although challenges and unanswered questions remain. There are many plausible mechanisms by which PFJ might be protective, and investigation of its effects on human health and disease prevention must remain an active area of research.


Brugada syndrome risk loci seem protective against atrial fibrillation.
Several studies have shown an overlap between genes involved in the pathophysiological mechanisms of atrial fibrillation (AF) and Brugada Syndrome (BrS). We investigated whether three single-nucleotide polymorphisms (SNPs) (rs11708996; G>C located intronic to SCN5A, rs10428132; T>G located in SCN10A, and rs9388451; T>C located downstream to HEY2) at loci associated with BrS in a recent genome-wide association study (GWAS) also were associated with AF. A total of 657 patients diagnosed with AF and a control group comprising 741 individuals free of AF were included. The three SNPs were genotyped using TaqMan assays. The frequencies of risk alleles in the AF population and the control population were compared in two-by-two models. One variant, rs10428132 at SCN10A, was associated with a statistically significant decreased risk of AF (odds ratio (OR)=0.77, P=0.001). A meta-analysis was performed by enriching the control population with allele frequencies from controls in the recently published BrS GWAS (2230 alleles). In this meta-analysis, both rs10428132 at SCN10A (OR=0.73, P=5.7 × 10(-6)) and rs11708996 at SCN5A (OR=0.80, P=0.02) showed a statistically significant decreased risk of AF. When assessing the additive effect of the three loci, we found that the risk of AF decreased in a dose-responsive manner with increasing numbers of risk alleles (OR=0.50, P=0.001 for individuals carrying ≥4 risk alleles vs ≤1 allele). In conclusion, the prevalence of three risk alleles previously associated with BrS was lower in AF patients than in patients free of AF, suggesting a protective role of these loci in developing AF.


"Presemantic" cognition in semantic dementia: six deficits in search of an explanation.
On the basis of a theory about the role of semantic knowledge in the recognition and production of familiar words and objects, we predicted that patients with semantic dementia would reveal a specific pattern of impairment on six different tasks typically considered "pre-" or "non-" semantic: reading aloud, writing to dictation, inflecting verbs, lexical decision, object decision, and delayed copy drawing. The prediction was that all tasks would reveal a frequency-by-typicality interaction, with patients performing especially poorly on lower-frequency items with atypical structure (e.g., words with an atypical spelling-to-sound relationship; objects with an atypical feature for their class, such as the hump on a camel, etc). Of 84 critical observations (14 patients performing 6 tasks), this prediction was correct in 84/84 cases; and a single component in a factor analysis accounted for 87% of the variance across seven measures: each patient's degree of impairment on atypical items in the six experimental tasks and a separate composite score reflecting his or her degree of semantic impairment. Errors also consistently conformed to the predicted pattern for both expressive and receptive tasks, with responses reflecting residual knowledge about the typical surface structure of each domain. We argue that these results cannot be explained as associated but unrelated deficits but instead are a principled consequence of a primary semantic impairment.


Examination of the transition state of the low-molecular mass small tyrosine phosphatase 1. Comparisons with other protein phosphatases.
The reactions of p-nitrophenyl phosphate (pNPP) with the low-molecular mass tyrosine phosphatase Stp1 and with the mutants D128N, D128A, D128E, and S18A have been studied by measurement of heavy-atom isotope effects in the substrate. The isotope effects were measured at the nonbridging oxygen atoms [18(V/K)nonbridge], at the bridging oxygen atom (the site of bond cleavage) [18(V/K)bridge], and at the nitrogen atom in the nitrophenol leaving group [15(V/K)]. The results with native Stp1 were 1.0160 +/- 0.0005 for 18(V/K)bridge, 1.0007 +/- 0.0001 for 15(V/K), and 1.0018 +/- 0.0003 for 18(V/K)nonbridge. The values for 18(V/K)nonbridge and 15(V/K) differ from those previously measured with other protein-tyrosine phosphatases and from those of the aqueous hydrolysis reaction of pNPP. The values indicate that in the transition state of the native Stp1 reaction the leaving group bears a partial negative charge, and there is nucleophilic interaction between the Cys nucleophile, and the phosphoryl group, causing some decrease in the nonbridge P-O bond order. The transition state remains highly dissociative with respect to the degree of bond cleavage to the leaving group. Mutation of the general acid from aspartic acid to glutamic acid slows catalysis but causes no change in the isotope effects and thus does not alter the degree of proton transfer to the leaving group in the transition state. Mutations of this residue to asparagine or alanine give values for 18(V/K)bridge of about 1.029, for 15(V/K) of about 1.003, and for 18(V/K)nonbridge of 1.0010 (D128A) to 1.0024 (D128N). These data indicate a dissociative transition state with the leaving group departing as the nitrophenolate anion and indicate more nucleophilic participation than in the aqueous hydrolysis of the pNPP dianion, just as in the native enzyme. The isotope effects with the S18A mutant, in which a hydrogen bonding stabilization of the anionic Cys nucleophile has been removed, were within experimental error of those with the native enzyme, indicating that this alteration has no effect on the transition state for phosphoryl transfer from pNPP.


Requirements of DFR1/Heartless, a mesoderm-specific Drosophila FGF-receptor, for the formation of heart, visceral and somatic muscles, and ensheathing of longitudinal axon tracts in CNS.
DFR1 encodes a mesoderm-specific fibroblast growth factor receptor in Drosophila. Here, we identified and characterized a protein-null mutant of DFR1 and examined DFR1 expression in embryos using anti-DFR1 antibody. Mutant phenotypes were completely rescued by a genomic fragment from the DFR1 locus. After invagination, mesodermal cells expressing DFR1 undergo proliferation and spread out dorsally to form a monolayer beneath the ectoderm. In mutant embryos, however, the mesoderm is not capable of extending to the normal dorsal limit and consequently mesodermal cells fail to receive ectodermal signals and thus rendered incapable of differentiating into primordia for the heart, visceral and somatic muscles. DFR1 is also required for normal development of the central nervous system. The absence of DFR1 resulted in the failure of longitudinal glia to enwrap longitudinal axon tracts. DFR1 mutant phenotypes were partially mimicked by the targeted expression of activated Yan, thus demonstrating the MAP kinase pathway to be involved in differentiation of mesoderm.


Behavioral deficits revealed by multiple tests in rats with ischemic damage limited to half of the CA1 sector of the hippocampus.
Ischemia causes neuronal necrosis in the septal half of the CA1 sector of the hippocampus in rats and produces deficits in spatial and other tasks, but the impairments are not severe. CA1-damaged rats may perform at control levels on some tests and may overlap the performance of control animals on other tests. The present study examined the performance of CA1-damaged rats on a range of tests known to be sensitive to hippocampal damage and evaluated the size of the deficit using a composite test-battery score. The rats with CA1 damage were impaired on learning set, matching to place, and latent-learning spatial navigation in a swimming pool, delayed alteration, and pattern alternation on dry land, but not on amphetamine-induced locomotion or on negative patterning. The CA1-damaged rats' impairments were mild, characterized by behavioral strategies similar to those of control rats, and their tests scores overlapped those of control rats. Nevertheless, a composite profile derived from the measures obtained from the different tests revealed a large overall impairment. The results confirm that rats with partial damage to CA1 display impairments, especially on tests that demand working memory, and suggest that the test-battery approach can be used to enhance group differences.


Comparison of Mayo and Milwaukee Risk Stratification Models for Predicting Lymph Node Metastasis in Endometrial Cancer.
The aim of this study was to compare Mayo and Milwaukee risk stratification models for predicting lymphatic dissemination in patients with endometrial cancer (EC). A total of 904 patients with EC underwent surgical treatment between 2004 and 2016 at Hacettepe University Hospital, and clinicopathological data of patients were retrieved from the computerized database of the Hacettepe University. Patients who did not undergo lymphadenectomy and who had nonendometrioid histology, stage-IV disease, and synchronous epithelial ovarian and EC were excluded. All slides of the cases were reviewed by the same gynecologic pathology subspecialist. The study group consisted of 307 consecutive patients with a mean age of 59.4 years (range, 26-86 years). Lymph node metastasis was detected in 28 subjects (9.1%). Primary tumor diameter, depth of myometrial invasion, lymphovascular space invasion, and cervical stromal and glandular involvement were associated with lymph node metastasis. Patients with low-risk histological features based on Mayo risk stratification system in our study group had a 0% rate of lymph node positivity. However, of the 28 patients with lymph node metastasis, 3 (10.7%) had low-risk features based on Milwaukee model. The sensitivity, specificity, false negative rate (FNR) and false positive rate of Mayo and Milwaukee risk stratification models for predicting lymphatic dissemination among women with endometrioid EC were 100%, 27.3%, 0%, and 72.7%; and 89.3%, 61.3%, 10.7%, and 38.7%, respectively. Although Milwaukee risk stratification model had a lower false positive rate and can decrease the number of lymphadenectomies, FNR of this new model was found as 10.7% in the present study. Furthermore, we found that Mayo model had a lower FNR and higher sensitivity. Therefore, Mayo model still looks more beneficial to predict lymph node metastasis in patients with endometrioid EC and Milwaukee risk stratification model still requires external validation.


Physical and psychosocial risk factors for musculoskeletal disorders in New Zealand nurses, postal workers and office workers.
To investigate the association of physical and psychosocial risk factors with musculoskeletal disorders (MSDs) in New Zealand nurses, postal workers and office workers. A cross-sectional postal survey asking about demographic, physical and psychosocial factors and MSDs. A total of 911 participants was randomly selected; nurses from the Nursing Council of New Zealand database (n=280), postal workers from their employer's database (n=280) and office workers from the 2005 electoral roll (n=351). Self-reported pain in the low back, neck, shoulder, elbow, wrist/hand or knee lasting more than 1 day in the month before the survey. The response rate was 58%, 443 from 770 potential participants. 70% (n=310) reported at least one MSDs. Physical work tasks were associated with low back (odds ratio (OR) 1.35, 95% CI 1.14 to 1.6), shoulder (OR 1.41, 95% CI 1.17 to 1.69), elbow (OR 1.14, 95% CI 1.13 to 1.83) and wrist/hand pain (OR 1.39, 95% CI 1.15 to 1.69). Job strain had the strongest association with neck pain (OR 3.46, 95% CI 1.30 to 9.21) and wrist/hand pain. Somatisation was weakly associated with MSDs at most sites. Better general and mental health status were weakly associated with lower odds of MSDs. In injury prevention and rehabilitation the physical nature of the work needs to be addressed for most MSDs, with modest decreases in risk seemingly possible. Addressing job strain could provide significant benefit for those with neck and wrist/hand pain, while the effects of somatisation and the promotion of good mental health may provide smaller but global benefits.


Disease Tolerance and Pathogen Resistance Genes May Underlie Trypanosoma cruzi Persistence and Differential Progression to Chagas Disease Cardiomyopathy.
Chagas disease is caused by infection with the protozoan Trypanosoma cruzi and affects over 8 million people worldwide. In spite of a powerful innate and adaptive immune response in acute infection, the parasite evades eradication, leading to a chronic persistent infection with low parasitism. Chronically infected subjects display differential patterns of disease progression. While 30% develop chronic Chagas disease cardiomyopathy (CCC)-a severe inflammatory dilated cardiomyopathy-decades after infection, 60% of the patients remain disease-free, in the asymptomatic/indeterminate (ASY) form, and 10% develop gastrointestinal disease. Infection of genetically deficient mice provided a map of genes relevant for resistance to T. cruzi infection, leading to the identification of multiple genes linked to survival to infection. These include pathogen resistance genes (PRG) needed for intracellular parasite destruction, and genes involved in disease tolerance (protection against tissue damage and acute phase death-DTG). All identified DTGs were found to directly or indirectly inhibit IFN-γ production or Th1 differentiation. We hypothesize that the absolute need for DTG to control potentially lethal IFN-γ PRG activity leads to T. cruzi persistence and establishment of chronic infection. IFN-γ production is higher in CCC than ASY patients, and is the most highly expressed cytokine in CCC hearts. Key DTGs that downmodulate IFN-γ, like IL-10, and Ebi3/IL27p28, are higher in ASY patients. Polymorphisms in PRG and DTG are associated with differential disease progression. We thus hypothesize that ASY patients are disease tolerant, while an imbalance of DTG and IFN-γ PRG activity leads to the inflammatory heart damage of CCC.


The subjective physiological, psychological, and behavioral risk-taking consequences of alcohol and energy drink co-ingestion.
The increasingly popular practice among adolescents and young adults of consuming alcohol mixed with energy drinks (AmED) has raised concern regarding potential increases in maladaptive drinking practices, negative psychological and physiological intoxication side effects, and risky behavioral outcomes. Comparison of user types has revealed that AmED users report engaging in more risk-taking behavior relative to alcohol users. However, the comparative likelihood of risk-taking according to session type (i.e., AmED vs. alcohol session) remains relatively unknown. Thus, this study was designed with the aim of establishing the subjective physiological, psychological, and behavioral risk-taking outcomes of AmED consumption relative to alcohol consumption for AmED users drawn from the community. Between May and June 2011, 403 Australians aged 18 to 35 who had consumed AmED and alcohol only in the preceding 6 months completed a 10- to 30-minute online survey about their use of these substances. Despite participants consuming a significantly greater quantity of alcohol in AmED sessions compared to alcohol sessions, the odds of participants experiencing disinhibition and engaging in 26 risk behaviors were significantly lower during AmED sessions relative to alcohol sessions. Similarly, the odds of experiencing several physiological (i.e., speech and walking difficulties, nausea, and slurred speech) and psychological (i.e., confusion, exhaustion, sadness) sedation outcomes were less during AmED sessions compared to alcohol sessions. However, the odds of enduring physiological (i.e., heart palpitations, sleep difficulties, agitation, tremors, jolt and crash episodes, and increased speech speed) and psychological (i.e., irritability and tension) outcomes potentially related to overstimulation were significantly greater during AmED sessions than alcohol sessions. Co-ingestion may provide a double-edged effect. The increased stimulation from energy drinks (EDs) may negate some intoxication-related sedation side effects by increasing alertness. However, it could also lead to negative physiological side effects associated with overstimulation. Notwithstanding any stimulatory effects of EDs, risk and negative effects of excessive alcohol consumption were present in both session types. However, the odds of engaging in risk-taking were less during AmED sessions relative to alcohol sessions. Objective measurement of behavioral risk-taking via laboratory-based measures could confirm the causal relationship between AmED and risk-taking.


Dynamic coronary roadmapping via catheter tip tracking in X-ray fluoroscopy with deep learning based Bayesian filtering.
Percutaneous coronary intervention (PCI) is typically performed with image guidance using X-ray angiograms in which coronary arteries are opacified with X-ray opaque contrast agents. Interventional cardiologists typically navigate instruments using non-contrast-enhanced fluoroscopic images, since higher use of contrast agents increases the risk of kidney failure. When using fluoroscopic images, the interventional cardiologist needs to rely on a mental anatomical reconstruction. This paper reports on the development of a novel dynamic coronary roadmapping approach for improving visual feedback and reducing contrast use during PCI. The approach compensates cardiac and respiratory induced vessel motion by ECG alignment and catheter tip tracking in X-ray fluoroscopy, respectively. In particular, for accurate and robust tracking of the catheter tip, we proposed a new deep learning based Bayesian filtering method that integrates the detection outcome of a convolutional neural network and the motion estimation between frames using a particle filtering framework. The proposed roadmapping and tracking approaches were validated on clinical X-ray images, achieving accurate performance on both catheter tip tracking and dynamic coronary roadmapping experiments. In addition, our approach runs in real-time on a computer with a single GPU and has the potential to be integrated into the clinical workflow of PCI procedures, providing cardiologists with visual guidance during interventions without the need of extra use of contrast agent.


Dissecting complex epigenetic alterations in breast cancer using CpG island microarrays.
It is now clear that aberrant DNA methylation observed in cancer cells is not restricted to a few CpG islands, but affects multiple loci. When this epigenetic event occurs at the 5'-end of the regulatory region of genes, it is frequently associated with transcriptional silencing. To investigate further this widespread event in the tumor genome, we developed a novel microarray containing 7776 short GC-rich tags tethered to glass slide surfaces. This DNA chip was used to study 17 paired tissues of breast tumors and normal controls. Amplicons, representing differential pools of methylated DNA fragments between tumors and normal controls, were cohybridized to the microarray panel. Hypermethylation of multiple CpG island loci was then detected in a two-color fluorescence system. Approximately 1% (on average, 83 loci) of these CpG islands examined were hypermethylated in this patient group. Hierarchical clustering segregated these tumors based on their methylation profiles and identified a group of CpG island loci that corresponds to the hormone-receptor status of breast cancer. This observation was independently confirmed by examining a single locus, the promoter of the human glypican 3 gene, which was predominately hypermethylated in the hormone receptor-negative tumors. Our findings support the notion that hypermethylation of critical CpG island loci influences cancer development and produces distinct epigenetic signatures for particular tumor subtypes.


Role of the atypical chemoattractant receptor CRAM in regulating CCL19 induced CCR7 responses in B-cell chronic lymphocytic leukemia.
The non-signalling chemokine receptors, including receptors DARC, D6 and CCX-CKR, have recently been shown to be involved in chemokine clearance and activity regulation. The human chemokine receptor CRAM (also known as HCR or CCRL2) is the most recently identified member of this atypical group. CRAM is expressed on B cells in a maturation-stage dependent manner and absent on T cells. We have recently shown that it competitively binds CCL19. CCL19 and its signalling receptor CCR7 are critical components involved in cell recruitment to secondary lymphoid organs and in maturation. B cell Chronic Lymphocytic Leukemia (B-CLL) is a low-grade lymphoma characterized by proliferative centres (or pseudofollicles). Proliferative centres develop due to abnormal cellular localisation and they are involved in the development of malignant cells. CCR7 is highly expressed on B cells from CLL patients and mediates migration towards its ligands CCL19 and CCL21, while CRAM expression and potential interferences with CCR7 are yet to be characterized. In this study, we show that B cells from patients with B-CLL present highly variable degrees of CRAM expression in contrast to more consistently high levels of CCR7. We investigated the hypothesis that, similar to the atypical receptor DARC, CRAM can modulate chemokine availability and/or efficacy, resulting in the regulation of cellular activation. We found that a high level of CRAM expression was detrimental to efficient chemotaxis with CCL19. MAP-kinase phosphorylation and intracellular calcium release induced by CCL19 were also altered by CRAM expression. In addition, we demonstrate that CRAM-induced regulation of CCL19 signalling is maintained over time. We postulate that CRAM is a factor involved in the fine tuning/control of CCR7/CCL19 mediated responses. This regulation could be critical to the pivotal role of CCL19 induced formation of proliferation centres supporting the T/B cells encounter as well as disease progression in B-CLL.


Plaque-prone hemodynamics impair endothelial function in pig carotid arteries.
Hemodynamic forces play an active role in vascular pathologies, particularly in relation to the localization of atherosclerotic lesions. It has been established that low shear stress combined with cyclic reversal of flow direction (oscillatory shear stress) affects the endothelial cells and may lead to an initiation of plaque development. The aim of the study was to analyze the effect of hemodynamic conditions in arterial segments perfused in vitro in the absence of other stimuli. Left common porcine carotid segments were mounted into an ex vivo arterial support system and perfused for 3 days under unidirectional high and low shear stress (6 +/- 3 and 0.3 +/- 0.1 dyn/cm(2)) and oscillatory shear stress (0.3 +/- 3 dyn/cm(2)). Bradykinin-induced vasorelaxation was drastically decreased in arteries exposed to oscillatory shear stress compared with unidirectional shear stress. Impaired nitric oxide-mediated vasodilation was correlated to changes in both endothelial nitric oxide synthase (eNOS) gene expression and activation in response to bradykinin treatment. This study determined the flow-mediated effects on native tissue perfused with physiologically relevant flows and supports the hypothesis that oscillatory shear stress is a determinant factor in early stages of atherosclerosis. Indeed, oscillatory shear stress induces an endothelial dysfunction, whereas unidirectional shear stress preserves the function of endothelial cells. Endothelial dysfunction is directly mediated by a downregulation of eNOS gene expression and activation; consequently, a decrease of nitric oxide production and/or bioavailability occurs.


Establishing an international laboratory network for neglected tropical diseases: Understanding existing capacity in five WHO regions.
Background. Limited laboratory capacity is a significant bottleneck in meeting global targets for the control and elimination of neglected tropical diseases (NTD). Laboratories are essential for providing clinical data and monitoring data about the status and changes in NTD prevalence, and for detecting early drug resistance. Currently NTD laboratory networks are informal and specialist laboratory expertise is not well publicised, making it difficult to share global expertise and provide training, supervision, and quality assurance for NTD diagnosis and research. This study aimed to identify laboratories within five World Health Organisation regions (South-East Asia, Eastern Mediterranean, Americas, Western Pacific and Europe) that provide NTD services and could be regarded as national or regional reference laboratories, and to conduct a survey to document their networks and capacity to support NTD programmes. Methods. Potential NTD reference laboratories were identified through systematic searches, snowball sampling and key informants. Results. Thirty-two laboratories responded to the survey. The laboratories covered 25 different NTDs and their main regional and national roles were to provide technical support and training, research, test validation and standard setting. Two thirds of the laboratories were based in academic institutions and almost half had less than 11 staff. Although greater than 90 per cent of the laboratories had adequate technical skills to function as an NTD reference laboratory, almost all laboratories lacked systems for external verification that their results met international standards. Conclusions. This study highlights that although many laboratories believed they could act as a reference laboratory, only a few had all the characteristics required to fulfil this role as they fell short in the standard and quality assurance of laboratory processes. Networks of high quality laboratories are essential for the control and elimination of disease and this study presents a critical first step in the development of such networks for NTDs.


A new in vivo Raman probe for enhanced applicability to the body.
This paper describes a new in vivo Raman probe that allows investigation of areas of the body that are otherwise difficult to access. It is coupled to a previously described commercially available in vivo Raman spectrometer that samples the skin through an optical flat. In the work presented here, the laser light emerges from a smaller pen-shaped probe. It thus works on the same principles as the original spectrometer, while its relative performance in terms of signal-to-noise ratio of the spectra and obtained spatial resolution is only slightly diminished. It allows the window to be placed against the subject in more curved and recessed areas of subject's body and also for them to be more comfortable while the measurements take place. Results from three areas of the body that have previously been very difficult to study are described, the mouth, axilla, and scalp. Results from the scalp and axilla strata cornea (SC) show significant differences from the "normal" SC of the volar forearm. For instance, the scalp is observed to have lower amounts of natural moisturizing factors (NMF) compared to the volar forearm within the same subjects. Also for both the axilla and scalp the lipids show a change in order as compared to the lipids in the volar forearm and also differences from each other. The potential significance of these observations is discussed. Further, we show how we can probe the mouth, in this case observing the presence of the astringent tea polyphenol epigallocatechin gallate within the oral mucosa.


Ectomycorrhizal specificity patterns in a mixed Pinus contorta and Picea engelmannii forest in Yellowstone National Park.
We used molecular genetic methods to test two hypotheses, (i) that host plant specificity among ectomycorrhizal fungi would be common in a closed-canopy, mixed Pinus contorta-Picea engelmannii forest in Yellowstone National Park and (ii) that specificity would be more common in the early successional tree species, P. contorta, than in the invader, P. engelmannii. We identified 28 ectomycorrhizal fungal species collected from 27 soil cores. The proportion of P. engelmannii to P. contorta ectomycorrhizae was nearly equal (52 and 48%, respectively). Of the 28 fungal species, 18 composed greater than 95% of the fungal community. No species was associated exclusively with P. contorta, but four species, each found in only one core, and one species found in two cores were associated exclusively with P. engelmannii. These fungi composed less than 5% of the total ectomycorrhizae. Thus, neither hypothesis was supported, and hypothesized benefits of ectomycorrhizal specificity to both trees and fungi probably do not exist in this system.


Accuracy of the energy expenditure during uphill exercise measured by the Waist-worn ActiGraph.
The application of Micro-Electro-Mechanical Sensors (MEMS) as measurements of energy expenditure (EE) has certain disadvantages. For example, the inertial sensors cannot easily distinguish changes in ground slope during walking/running conditions, so the accuracy of EE calculation is biased. To resolve this issue, heart rate (HR) and heart rate reserve (HRR) were used as compensatory factors respectively to correct the classical empirical formula of the accelerometer analyzer for EE in this study. To explore the improvement of the accuracy of EE during uphill exercise and compare the correction levels between HR and HRR, oxygen uptake was used as a criterion measure (CM). Thirty healthy adult males wore an ActiGraph GT3X with the Polar HR monitor and Vmax indirect calorimeter during twelve treadmill activities (3 gradients and 4 speeds). When the slopes were increased by 0%, 3%, and 6%, the measurement accuracy of the accelerometers, calculated by intraclass correlation coefficient (ICC), decreased by 0.877, 0.755, and 0.504, respectively (p < 0.05). The HR and HRR parameters of linear regression were used to correct the classical formula. The results showed that HR had higher coefficients of determination (R2) (0.801, 0.700, and 0.642 respectively) and ICCs (0.887, 0.825, and 0.785 respectively) than did the accelerometer outputs. HRR showed the highest coefficients of determination (R2) (0.821, 0.728, and 0.656 respectively) and ICCs (0.901, 0.844, and 0.795 respectively). Through adding HRR parameters, the accuracy of the classical prediction formula EE was significantly improved during walking/running on sloping ground.


Dominant colonization and inheritance of Methylobacterium sp. strain OR01 on perilla plants.
Pink-pigmented facultative methylotrophs (PPFMs) are major inhabitants of the phyllosphere. In a preceding study, we found that perilla plants harbor a dominant population of PPFMs on their leaves and seeds, and that the closest relative of PPFMs (Methylobacterium sp. strain OR01 as representative strain) isolated from red perilla seeds was M. fujisawaense DSM5686(T). In the present study, the specific interaction between red perilla and Methylobacterium species was investigated. All the PPFMs isolated from red perilla seeds harvested in the Ohara area of Kyoto, Japan in 2009, 2010, and 2011 and the PPFMs isolated from red perilla leaves planted at four geographically different places in Japan had 16S rRNA sequences identical to that of strain OR01. Direct transmission of PPFMs from seeds to leaves and the competitiveness of strain OR01 were confirmed. This report is the first step toward understanding the species-level specificity of the interaction between perilla plants and Methylobacterium species.


Variability in the costs of institutional review board oversight.
Previous studies have shown wide differences between institutions in economies of scale with regard to the costs of institutional review board (IRB) oversight of research. In this study, the authors explored variability among IRB costs, taking into account organizational size, components of the costs of oversight, and protocol type. The authors conducted a survey of academic medical centers to collect information on resource utilization associated with IRB oversight in 2002. They used national cost weights to assign a cost to each type of resource used, and summed weighted resource utilization for IRB costs. Descriptive statistics were generated for costs over all, tertile of protocol volume, cost component, and type of review. They also determined where the greatest cost variability is found. IRB costs per protocol reviewed are highly variable both overall and within tertiles of volume. Higher-volume institutions have lower costs, which is indicative of economies of scale. However, not all components of IRB costs (e.g., board time) are subject to economies of scale. Expedited reviews of protocols are not less expensive at low-volume institutions. IRB costs for oversight are highly variable, and only some of the variation may be attributable to economies of scale. Given such wide variation in costs, the authors conclude that some institutions are conducting reviews in a manner that is inefficient or of low quality. Future work is needed to determine specific practices in reviews, and what leads to the best quality and most efficient oversight and review system.


Pulmonary Sarcomatoid Carcinomas Commonly Harbor Either Potentially Targetable Genomic Alterations or High Tumor Mutational Burden as Observed by Comprehensive Genomic Profiling.
Pulmonary sarcomatoid carcinoma (PSC) is a high-grade NSCLC characterized by poor prognosis and resistance to chemotherapy. Development of targeted therapeutic strategies for PSC has been hampered because of limited and inconsistent molecular characterization. Hybrid capture-based comprehensive genomic profiling was performed on DNA from formalin-fixed paraffin-embedded sections of 15,867 NSCLCs, including 125 PSCs (0.8%). Tumor mutational burden (TMB) was calculated from 1.11 megabases (Mb) of sequenced DNA. The median age of the patients with PSC was 67 years (range 32-87), 58% were male, and 78% had stage IV disease. Tumor protein p53 gene (TP53) genomic alterations (GAs) were identified in 74% of cases, which had genomics distinct from TP53 wild-type cases, and 62% featured a GA in KRAS (34%) or one of seven genes currently recommended for testing in the National Comprehensive Cancer Network NSCLC guidelines, including the following: hepatocyte growth factor receptor gene (MET) (13.6%), EGFR (8.8%), BRAF (7.2%), erb-b2 receptor tyrosine kinase 2 gene (HER2) (1.6%), and ret proto-oncogene (RET) (0.8%). MET exon 14 alterations were enriched in PSC (12%) compared with non-PSC NSCLCs (∼3%) (p < 0.0001) and were more prevalent in PSC cases with an adenocarcinoma component. The fraction of PSC with a high TMB (>20 mutations per Mb) was notably higher than in non-PSC NSCLC (20% versus 14%, p = 0.056). Of nine patients with PSC treated with targeted or immunotherapies, three had partial responses and three had stable disease. Potentially targetable GAs in National Comprehensive Cancer Network NSCLC genes (30%) or intermediate or high TMB (43%, >10 mutations per Mb) were identified in most of the PSC cases. Thus, the use of comprehensive genomic profiling in clinical care may provide important treatment options for a historically poorly characterized and difficult to treat disease.


Intravital Assessment of Cells Responses to Conducting Polymer-Coated Carbon Microfibres for Bridging Spinal Cord Injury.
The extension of the lesion following spinal cord injury (SCI) poses a major challenge for regenerating axons, which must grow across several centimetres of damaged tissue in the absence of ordered guidance cues. Biofunctionalized electroconducting microfibres (MFs) that provide biochemical signals, as well as electrical and mechanical cues, offer a promising therapeutic approach to help axons overcome this blind journey. We used poly(3,4-ethylenedioxythiophene)-coated carbon MFs functionalized with cell adhesion molecules and growth factors to bridge the spinal cord after a partial unilateral dorsal quadrant lesion (PUDQL) in mice and followed cellular responses by intravital two-photon (2P) imaging through a spinal glass window. Thy1-CFP//LysM-EGFP//CD11c-EYFP triple transgenic reporter animals allowed real time simultaneous monitoring of axons, myeloid cells and microglial cells in the vicinity of the implanted MFs. MF biocompatibility was confirmed by the absence of inflammatory storm after implantation. We found that the sprouting of sensory axons was significantly accelerated by the implantation of functionalized MFs after PUDQL. Their implantation produced better axon alignment compared to random and misrouted axon regeneration that occurred in the absence of MF, with a most striking effect occurring two months after injury. Importantly, we observed differences in the intensity and composition of the innate immune response in comparison to PUDQL-only animals. A significant decrease of immune cell density was found in MF-implanted mice one month after lesion along with a higher ratio of monocyte-derived dendritic cells whose differentiation was accelerated. Therefore, functionalized carbon MFs promote the beneficial immune responses required for neural tissue repair, providing an encouraging strategy for SCI management.


Phase I and pharmacokinetic study of photodynamic therapy for high-grade gliomas using a novel boronated porphyrin.
To determine the recommended dose, toxicity profile, and pharmacokinetics of a novel boronated porphyrin (BOPP) for photodynamic therapy (PDT) of intracranial tumors. BOPP was administered alone in increasing doses (0.25, 0.5, 1.0, 2.0, 4.0, or 8.0 mg/kg) preoperatively in patients with intracranial tumors undergoing postresection PDT until dose-limiting toxicity (DLT) was observed. Twenty-nine assessable patients with intracranial tumors received BOPP intravenously 24 hours before surgery. The recommended dose was 4 mg/kg. Dose escalation was limited by thrombocytopenia. The most common nonhematologic toxicity was skin photosensitivity. Pharmacokinetic parameters showed increased area under the plasma concentration-time curve and maximum concentration with increased dose. Tumor BOPP concentrations also increased with increased dose. BOPP at a dose of 4 mg/kg was well tolerated. DLT was thrombocytopenia, and photosensitivity was the only other toxicity of note. The efficacy of PDT using BOPP requires further exploration.


Mussel-inspired immobilization of vascular endothelial growth factor (VEGF) for enhanced endothelialization of vascular grafts.
Most polymeric vascular prosthetic materials have low patency rate for replacement of small diameter vessels (<5 mm), mainly due to failure to generate healthy endothelium. In this study, we present polydopamine-mediated immobilization of growth factors on the surface of polymeric materials as a versatile tool to modify surface characteristics of vascular grafts potentially for accelerated endothelialization. Polydopamine was deposited on the surface of biocompatible poly(L-lactide-co-ε-caprolactone) (PLCL) elastomer, on which vascular endothelial growth factor (VEGF) was subsequently immobilized by simple dipping. Surface characteristics and composition were investigated by using scanning electron microscopy, atomic force microscopy, and X-ray photoelectron spectroscopy. Immobilization of VEGF on the polydopamine-deposited PLCL films was effective (19.8 ± 0.4 and 197.4 ± 19.7 ng/cm(2) for DPv20 and DPv200 films, respectively), and biotin-mediated labeling of immobilized VEGF revealed that the fluorescence intensity increased as a function of the concentration of VEGF solution. The effect of VEGF on adhesion of HUVECs was marginal, which may have been masked by polydopamine layer that also enhanced cell adhesion. However, VEGF-immobilized substrate significantly enhanced proliferation of HUVECs for over 7 days of in vitro culture and also improved their migration. In addition, immobilized VEGF supported robust cell to cell interactions with strong expression of CD 31 marker. The same process was effective for immobilization of basic fibroblast growth factor, demonstrating the robustness of polydopamine layer for secondary ligation of growth factors as a simple and novel surface modification strategy for vascular graft materials.


Enhancing Change-of-Direction Speed in Soccer Players by Functional Inertial Eccentric Overload and Vibration Training.
To examine the effects of a novel isoinertial eccentric-overload and vibration training (EVT) paradigm on change-of-direction (COD) speed and multiple performance tests applicable to soccer. Twenty-four young male players were assigned to an EVT (n = 12) or conventional combined (CONV, n = 12) group, once weekly for 11 wk. EVT consisted of 2 sets of 6-10 repetitions in 5 specific and 3 complementary exercises. CONV used comparable volume (2 sets of 6-10 reps in 3 sequences of 3 exercises) of conventional combined weight, plyometric, and linear speed exercises. Pre- and postintervention tests included 25-m sprint with 4 × 45° COD every 5th m (V-cut test), 10- and 30-m sprints, repeat-sprint ability, countermovement jump, and hopping (RJ5). Group comparison showed very likely to likely better performance for EVT in the COD (effect size [ES] = 1.42), 30-m (ES = 0.98), 10-m (ES = 1.17), and average power (ES = 0.69) and jump height (ES = 0.69) during RJ5. There was a large (r = -.55) relationship between the increase in average hopping power and the reduced V-cut time. As EVT, not CONV, improved not only COD ability but also linear speed and reactive jumping, this "proof-of-principle" study suggests that this novel exercise paradigm performed once weekly could serve as a viable adjunct to improve performance tasks specific to soccer.


L-Arginine: a novel therapy for coronary artery disease?
Coronary artery disease (CAD) is a significant cause of morbidity and mortality today. The treatment of CAD is improving, but its prevalence is increasing: both primary and secondary prevention measures are of vital importance. Atherosclerosis starts at an early age; it is initiated at the vascular endothelium level, a single layer entity that modulates vascular function. Modulation of vascular function is carried out through the L-arginine/nitric oxide (NO) pathway. Normal endothelial function requires an intact L-arginine/NO pathway and endothelium. Endothelial dysfunction may be a precursor to overt CAD. CAD risk factors have been shown to influence endothelial function, and the treatment of these risk factors can restore endothelial function. L-Arginine is a safe, novel, semiessential amino acid that increases NO production, thereby improving endothelial function. L-Arginine/NO has numerous beneficial neurohormonal modulating properties. Numerous animal model and human studies have been carried out to assess L-arginine in CAD and other related disorders such as congestive heart failure (CHF), peripheral vascular disease (PVD) and acute myocardial infarction (AMI). Prospective clinical trials are required to assess the promising role of L-arginine in CAD and related disorders


Neuronal nitric oxide synthase regulates basal microvascular tone in humans in vivo.
Nitric oxide (NO) has a pivotal role in the regulation of vascular tone and blood flow, with dysfunctional release contributing to disease pathophysiology. These effects have been attributed to NO production by the endothelial NO synthase (eNOS); however, recent evidence suggests that a neuronal NO synthase (nNOS) may also be expressed in arterial vessels. We undertook a first-in-humans investigation of the role of nNOS in the local regulation of vascular blood flow in healthy subjects. Brachial artery infusion of the nNOS-specific inhibitor S-methyl-L-thiocitrulline (SMTC, 0.025 micromol/min to 0.2 micromol/min) caused a dose-dependent reduction in basal flow, with a 30.1+/-3.8% decrease at the highest dose (n=10; mean+/-SE; P<0.01). The effect of SMTC was abolished by coinfusion of the NO synthase substrate L-arginine but was unaffected by D-arginine. A similar reduction in basal flow with the nonselective NO synthase inhibitor N(G)-monomethyl-L-arginine (L-NMMA; 37.4+/-3.1%, n=10) required a 20-fold higher dose of 4 micromol/min. At doses that produced comparable reductions in basal flow, only L-NMMA (4 micromol/min) and not SMTC (0.2 micromol/min) inhibited acetylcholine-induced vasodilation; however, both SMTC and L-NMMA inhibited the forearm vasodilator response to mental stress. Basal forearm blood flow in humans is regulated by nNOS-derived NO, in contrast to the acetylcholine-stimulated increase in blood flow, which, as shown previously, is mediated primarily by eNOS. These data indicate that vascular nNOS has a distinct local role in the physiological regulation of human microvascular tone in vivo.


In vitro photo-controlled drug release system based on amphiphilic linear-dendritic diblock copolymers; self-assembly behavior and application as nanocarrier.
A simple type of photoresponsive amphiphilic linear-dendritic diblock copolymer has been synthesized and investigated for its ability to act as a drug carrier. These structures contain hydrophilic polyethylene oxide monomethyl ether (PEOM) as hydrophilic block and carbosiloxane dendritic branches as hydrophobic block grafted by two and six Magneson II as azo chromophore, PEOM-Azo, 2 and PEOM-Azo, 6 respectively. Self-assembling of the amphiphilic macromolecules of PEOM-Azo, 2 and PEOM-Azo, 6, briefly were represented as PEOM-Azo [2, 6], leads to the formation of their micellar aggregates in aqueous media. Their micellar properties such as critical micelle concentration (CMC), aggregation number and thereby total numbers of azo-groups in each micellar aggregates were determined. Also, they were characterized by TEM, SEM and DLS. The unloaded aggregates examined under UV light (366 nm), which were observed to be smaller than 300 nm. The release patterns of the encapsulated drug molecule from these aggregates were investigated as regulated by the PEOM-Azo [2, 6] systems in trans and cis forms. A comparison of the release behavior of the loaded PEOM-Azo [2, 6] systems indicated that the release rate of the encapsulated active molecules from the carriers was slower when the azo moieties were in trans form as compared to that the azo in the cis form. The in vitro release behavior of drug from these polymeric systems represents potential of the carriers for controlled drug delivery.


Fluorinert, an oxygen carrier, improves cell culture performance in deep square 96-well plates by facilitating oxygen transfer.
In bioprocess development, the 96-well plate format has been widely used for high-throughput screening of production cell line or culture conditions. However, suspension cell cultures in conventional 96-well plates often fail to reach high cell density under normal agitation presumably due to constraints in oxygen transfer. Although more vigorous agitation can improve gas transfer in 96-well plate format, it often requires specialized instruments. In this report, we employed Fluorinert, a biologically inert perfluorocarbon, to improve oxygen transfer in 96-well plate and to enable the growth of a Chinese Hamster Ovary cell line expressing a recombinant monoclonal antibody. When different amounts of Fluorinert were added to the cell culture medium, a dose-dependent improvement in cell growth was observed in both conventional and deep square 96-well plates. When sufficient Fluorinert was present in the culture, the cell growth rate, the peak cell density, and recombinant protein production levels achieved in deep square 96-wells were comparable to cultures in ventilated shake flasks. Although Fluorinert is known to dissolve gases such as oxygen and CO(2), it does not dissolve nor extract medium components, such as glucose, lactate, or amino acids. We conclude that mixing Fluorinert with culture media is a suitable model for miniaturization of cell line development and process optimization. Proper cell growth and cellular productivity can be obtained with a standard shaker without the need for any additional aeration or vigorous agitation.


Gait in attention deficit hyperactivity disorder : effects of methylphenidate and dual tasking.
Cognitive function and the loading of attention presumably play an important role in gait as well as in fall risk, but previous work has not demonstrated this in any cause-and-effect way. To gain insight into the relationship between gait and cognitive function, we sought: (1) To compare the gait rhythmicity (stride time variability) of children with attention deficit hyperactivity disorder (ADHD) to controls, (2) To test the hypothesis that dual tasking leads to increased stride-to-stride variability in ADHD, and (3) To test whether pharmacological treatment that relieves ADHD symptoms reduces stride-to-stride variability. Gait was quantified in children with ADHD and in age-matched healthy controls under single task and dual task conditions on three occasions: off medications (both groups) and, in the ADHD group, after double blinded, randomized administration of methylphenidate (MPH) or placebo. At baseline, children with ADHD tended to walk with increased stride-to-stride variability compared to the controls during the single task condition (p = 0.09). During dual task walking, stride time variability was significantly reduced in the children with ADHD (p < 0.004), but not in the controls. In the children with ADHD, the placebo did not significantly affect stride-to-stride variability or the dual tasking response. In contrast, stride time variability was significantly reduced on MPH (p < 0.001) such that dual tasking no longer affected variability. The present findings demonstrate alterations in the gait of children with ADHD, support a cause and effect link between cognitive function and gait, and suggest that enhancement of attention abilities may, in certain populations, improve gait rhythmicity.


Effects of China's current Air Pollution Prevention and Control Action Plan on air pollution patterns, health risks and mortalities in Beijing 2014-2018.
Beijing is one of the most polluted cities in the world. However, the "Air Pollution Prevention and Control Action Plan" (APPCAP), introduced since 2013 in China, has created an unprecedented drop in pollution concentrations for five major pollutants, except O3, with a significant drop in mortalities across most parts of the city. To assess the effects of APPCAP, air pollution data were collected from 35 sites (divided into four types, namely, urban, suburban, regional background, and traffic) in Beijing, from 2014 to 2018 and analyzed. Simultaneously, health-risk based air quality index (HAQI) and district-specific pollution (PM2.5 and O3) attributed mortality were calculated for Beijing. The results show that the annual PM2.5 concentration exceeded the Chinese national ambient air quality standard Grade II (35 μg/m3) in all sites, ranging from 88.5 ± 77.4 μg/m3 for the suburban site to 98.6 ± 89.0 μg/m3 for the traffic site in 2014, but was reduced to 50.6 ± 46.6 μg/m3 for the suburban site, and 56.1 ± 47.0 μg/m3 for the regional background in 2018. O3 was another most important pollutant that exceeded the Grade II standard (160 μg/m3) for a total of 291 days. It peaked at 311.6 μg/m3 in 2014 for the urban site and 290.6 μg/m3 in 2018 in the suburban site. APPCAP led to a significant reduction in PM2.5, PM10, NO2, SO2 and CO concentrations by 7.4, 8.1, 2.4, 1.9 and 80 μg/m3/year respectively, though O3 concentration was increased by 1.3 μg/m3/year during the five-years. HAQI results suggest that during the high pollution days, the more vulnerable groups, such as the children, and the elderly, should take additional precautions, beyond the recommendations currently put forward by Beijing Municipal Environmental Monitoring Center (BJMEMC). In 2014, PM2.5 and O3 attributed to 29,270 and 3,030 deaths respectively, though in 2018 their mortalities were reduced by 5.6% and 18.5% respectively. The highest mortality was observed in Haidian and Chaoyang districts, two of the most densely populated areas in Beijing. Beijing's air quality has seen a dramatic improvement over the five-year period, which can be attributable to the implementation of APPCAP and the central government's determination, with significant drops in the mortalities due to PM2.5 and O3 in parallel. To further improve air quality in Beijing, more stringent regulatory measures should be introduced to control volatile organic compounds (VOCs) and reduce O3 concentrations. Consistent air pollution control interventions will be needed to ensure long-term prosperity and environmental sustainability in Beijing, China's most powerful city. This study provides a robust methodology for analyzing air pollution trends, health risks and mortalities in China. The crucial evidence generated forms the basis for the governments in China to introduce location-specific air pollution policy interventions to further reduce air pollution in Beijing and other parts of China. The methodology presented in this study can form the basis for future fine-grained air pollution and health risk study at the city-district level in China.


Protein and steroid profiles in follicular fluid after ovarian hyperstimulation as potential biomarkers of IVF outcome.
Controlled ovarian hyperstimulation is performed to assist with generation of multiple mature oocytes for use in in vitro fertilization (IVF). The goal of our study was to evaluate differences in protein and steroid profiles in ovarian follicular fluid (hFF) samples obtained during oocyte retrieval from women undergoing IVF treatment and to identify physiological pathways associated with the proteins. The hFF samples were depleted of abundant proteins, fractionated by ultrafiltration, digested, and analyzed by nano-LC-QTOF. Concentrations of 15 endogenous steroids were determined in the samples using LC-MS/MS methods. The total number of proteins identified in the samples was 75, of which 4, 7, and 2 were unique to the samples from women with viable pregnancy, miscarriage, and no pregnancy, respectively. Identified proteins were associated with the acute response signaling, coagulation system, intrinsic and extrinsic prothrombin activation, complement system, neuroprotective role of THOP1, FXR/RXR activation, role of tissue factor, and growth hormone pathways. A greater number of proteins associated with biosynthesis was found in hFF samples corresponding to the oocytes resulting in pregnancy. The abundance of seven proteins was found to be associated with steroidogenesis. The obtained data will contribute to better understanding of the pathogenesis and development of noninvasive markers for assessment of oocytes viability.


Screening for pulmonary arterial hypertension in patients with systemic sclerosis: clinical characteristics at diagnosis and long-term survival.
Pulmonary arterial hypertension (PAH) is a severe, life-limiting complication of systemic sclerosis (SSc). Guidelines recommend early detection and management of SSc-PAH. However, little is known about the impact of detection programs on patients with SSc-PAH. This study was undertaken to assess the clinical characteristics of patients with SSc-PAH at diagnosis and their long-term outcomes. Two incident cohorts of patients with SSc-PAH from the same management era (2002/2003) were studied. The first cohort (designated the routine practice cohort) included consecutive adult patients with symptomatic SSc in whom a diagnosis of PAH was made by right-sided heart catheterization (RHC) at the time of recruitment into the French PAH Registry. The second cohort (designated the detection cohort) comprised consecutive patients with SSc who entered a systematic PAH detection program and were subsequently found to have PAH on RHC. Clinical characteristics at diagnosis of PAH and subsequent 8-year mortality were compared between the cohorts. There were 16 patients in each cohort. At the time of PAH diagnosis, patients in the detection cohort had less advanced pulmonary vascular disease compared with patients in the routine practice cohort, as evidenced by more patients being in New York Heart Association class I and class II, a lower mean pulmonary artery pressure and pulmonary vascular resistance index, and a higher cardiac output. Patients in the detection cohort were less likely to receive diuretics and warfarin, but there was no difference in exposure to PAH-specific therapies between the cohorts. The 1-, 3-, 5-, and 8-year survival rates were 75%, 31%, 25%, and 17%, respectively, in the routine practice cohort compared with 100%, 81%, 73%, and 64%, respectively, in the detection cohort (P = 0.0037). Compared with patients in routine clinical practice, PAH detection programs in SSc are able to identify patients with milder forms of the disease, allowing earlier management.