Performance analysis of efforts towards promotion of corneal donation at a tertiary care trauma center in India.
To identify the number of potential donors from trauma-related deaths and examine the extent of loss of opportunity at different levels for eye donation. A retrospective review of the death records from the mortuary of Jai Prakash Narayan Apex Trauma Centre and National Eye Bank at All India Institute of Medical Sciences was conducted for the period between January 2008 and August 2009. All cases with trauma-related deaths were included in the study. Cases with known contraindications to eye donation were excluded from the study. The main outcome measure was "lost opportunity cases" where the families of eligible donors were not approached. The secondary outcome measure was successful procurement rate. The total number of trauma-related deaths noted during the study period was 1584. One hundred cases were excluded from the study because the records were either incomplete or not readily available. A total of 1066 cases were identified as medically suitable potential eye donors. The death-autopsy interval was 12 hours or less in 425 cases and >12 hours in 641 cases. Among eligible donors, there were 831 (78%) lost opportunity cases. Among 235 families approached, only 20 (8.5%) agreed to eye donation. Overall, successful eye donation was possible in only 20 (1.9%) of the eligible trauma-related deaths. Trauma-related deaths form a major pool for potential tissue donors. Only about one fifth of the medically eligible cases could be approached for counseling. A potentially large donor pool of trauma-related deaths is compromised by the lost opportunity cases and the low donation rate. Efforts are needed to augment existing administrative and manpower resources to increase the corneal procurement rate in the Indian population.


Ethnic disparities in self-reported oral health status and access to care among older adults in NYC.
There is a growing burden of oral disease among older adults that is most significantly borne by minorities, the poor, and immigrants. Yet, national attention to oral heath disparities has focused almost exclusively on children, resulting in large gaps in our knowledge about the oral health risks of older adults and their access to care. The projected growth of the minority and immigrant elderly population as a proportion of older adults heightens the urgency of exploring and addressing factors associated with oral health-related disparities. In 2008, the New York City Health Indicators Project (HIP) conducted a survey of a representative sample of 1,870 adults over the age of 60 who attended a random selection of 56 senior centers in New York City. The survey included questions related to oral health status. This study used the HIP database to examine differences in self-reported dental status, dental care utilization, and dental insurance, by race/ethnicity, among community-dwelling older adults. Non-Hispanic White respondents reported better dental health, higher dental care utilization, and higher satisfaction with dental care compared to all other racial/ethnic groups. Among minority older adults, Chinese immigrants were more likely to report poor dental health, were less likely to report dental care utilization and dental insurance, and were less satisfied with their dental care compared to all other racial/ethnic groups. Language fluency was significantly related to access to dental care among Chinese immigrants. Among a diverse community-dwelling population of older adults in New York City, we found significant differences by race/ethnicity in factors related to oral health. Greater attention is needed in enhancing the cultural competency of providers, addressing gaps in oral health literacy, and reducing language barriers that impede access to care.


Laser-induced dispersion control.
An intense laser pulse is used to control the spectral phase of a weak probe pulse as they overlap in fused silica. The laser-induced linear chirp is controlled by the delay time between pulses. Dependence from intensity and spectral phase of the pump pulse is also studied. Experimental data is validated by numerical simulation based on optical Kerr effect. Results show that laser-induced pulse shaping is possible and may be useful for intracavity pulse compression and shaping in enhancement cavities.


Allosteric interactions required for high-affinity binding of dihydropyridine antagonists to Ca(V)1.1 Channels are modulated by calcium in the pore.
Dihydropyridines (DHPs) are an important class of drugs, used extensively in the treatment of angina pectoris, hypertension, and arrhythmia. The molecular mechanism by which DHPs modulate Ca(2+) channel function is not known in detail. We have found that DHP binding is allosterically coupled to Ca(2+) binding to the selectivity filter of the skeletal muscle Ca(2+) channel Ca(V)1.1, which initiates excitation-contraction coupling and conducts L-type Ca(2+) currents. Increasing Ca(2+) concentrations from approximately 10 nM to 1 mM causes the DHP receptor site to shift from a low-affinity state to a high-affinity state with an EC(50) for Ca(2+) of 300 nM. Substituting each of the four negatively charged glutamate residues that form the ion selectivity filter with neutral glutamine or positively charged lysine residues results in mutant channels whose DHP binding affinities are decreased up to 10-fold and are up to 150-fold less sensitive to Ca(2+) than wild-type channels. Analysis of mutations of amino acid residues adjacent to the selectivity filter led to identification of Phe-1013 and Tyr-1021, whose mutation causes substantial changes in DHP binding. Thermo-dynamic mutant cycle analysis of these mutants demonstrates that Phe-1013 and Tyr-1021 are energetically coupled when a single Ca(2+) ion is bound to the channel pore. We propose that DHP binding stabilizes a nonconducting state containing a single Ca(2+) ion in the pore through which Phe-1013 and Tyr-1021 are energetically coupled. The selectivity filter in this energetically coupled high-affinity state is blocked by bound Ca(2+), which is responsible for the high-affinity inhibition of Ca(2+) channels by DHP antagonists.


Peridomestic risk factors for canine leishmaniasis in urban dwellings: new findings from a prospective study in Brazil.
Despite the wealth of information on the prevalence and correlates of canine Leishmania infection (CLI), data on its incidence are still sparse, and little is known regarding risk factors for CLI. We studied a cohort of dogs in an urban area in Brazil to determine whether incidence varied with age, breed, and environmental characteristics. The mean follow-up was 1.5 years, and the crude incidence rate was 11.8 cases/100 dog-years (95% confidence interval [CI] = 8.6-15.6). In the multivariate analysis, short fur was the strongest predictor of CLI (relative risk [RR] = 9.4). In addition, our data indicate that raising pigs (RR = 4.1), chickens (RR = 3.3), or other livestock (RR = 2.6) significantly increased the risk of CLI. Thus, suggesting control measures directed towards modifying the environmental factors favoring contact between vectors, reservoirs, and susceptible humans, such as proximity to pigpens or hen houses. Furthermore, conventional control programs of insecticidal spraying of human dwellings should also apply insecticide in and around animal sheds.


Determinants of contraceptive method choice in rural Tanzania between 1991 and 1999.
Four pooled Demographic and Health Survey data sets are used to examine the determinants of contraceptive method choice in rural Tanzania for the period from 1991 to 1999. The individual data are linked to facility surveys conducted in the same communities so that the impact of Tanzania's family planning program can be examined. The focus of the study is an examination of the effect on method choice of the three major components of Tanzania's family planning program: logistical support, trained providers, and communications programs. The statistical methods employed correct for the potential endogeneity of family planning message recall. Simulations are used to quantify the impact of the important policy variables. All three components of the program are shown to have had an impact on modern method choice.


Two to Tango: Dialog between Immunity and Stem Cells in Health and Disease.
Stem cells regenerate tissues in homeostasis and under stress. By taking cues from their microenvironment or "niche," they smoothly transition between these states. Immune cells have surfaced as prominent members of stem cell niches across the body. Here, we draw parallels between different stem cell niches to explore the context-specific interactions that stem cells have with tissue-resident and recruited immune cells. We also highlight stem cells' innate ability to sense and respond to stress and the enduring memory that forms from such encounters. This fascinating crosstalk holds great promise for novel therapies in inflammatory diseases and regenerative medicine.


Interactions between β-catenin and the HSlo potassium channel regulates HSlo surface expression.
The large conductance calcium-activated potassium channel alpha-subunit (Slo) is widely distributed throughout the body and plays an important role in a number of diseases. Prior work has shown that Slo, through its S10 region, interacts with β-catenin, a key component of the cytoskeleton framework and the Wnt signaling pathway. However, the physiological significance of this interaction was not clear. Using a combination of proteomic and cell biology tools we show the existence of additional multiple binding sites in Slo, and explore in detail β-catenin interactions with the S10 region. We demonstrate that deletion of this region reduces Slo surface expression in HEK cells, which indicates that interaction with beta-catenin is important for Slo surface expression. This is confirmed by reduced expression of Slo in HEK cells and chicken (Gallus gallus domesticus leghorn white) hair cells treated with siRNA to β-catenin. HSlo reciprocally co-immunoprecipitates with β-catenin, indicating a stable binding between these two proteins, with the S10 deletion mutant having reduced binding with β-catenin. We also observed that mutations of the two putative GSK phosphorylation sites within the S10 region affect both the surface expression of Slo and the channel's voltage and calcium sensitivities. Interestingly, expression of exogenous Slo in HEK cells inhibits β-catenin-dependent canonical Wnt signaling. These studies identify for the first time a central role for β-catenin in mediating Slo surface expression. Additionally we show that Slo overexpression can lead to downregulation of Wnt signaling.


Representing perturbed dynamics in biological network models.
We study the dynamics of gene activities in relatively small size biological networks (up to a few tens of nodes), e.g., the activities of cell-cycle proteins during the mitotic cell-cycle progression. Using the framework of deterministic discrete dynamical models, we characterize the dynamical modifications in response to structural perturbations in the network connectivities. In particular, we focus on how perturbations affect the set of fixed points and sizes of the basins of attraction. Our approach uses two analytical measures: the basin entropy H and the perturbation size Delta , a quantity that reflects the distance between the set of fixed points of the perturbed network and that of the unperturbed network. Applying our approach to the yeast-cell-cycle network introduced by Li [Proc. Natl. Acad. Sci. U.S.A. 101, 4781 (2004)] provides a low-dimensional and informative fingerprint of network behavior under large classes of perturbations. We identify interactions that are crucial for proper network function, and also pinpoint functionally redundant network connections. Selected perturbations exemplify the breadth of dynamical responses in this cell-cycle model.


Effect of administering cyclophosphamide and vitamin E on the levels of tumor-marker enzymes in rats with experimentally induced fibrosarcoma.
Cyclophosphamide, and antineoplastic drug, and vitamin E, the common antioxidant present in the diet, were administered in separate dosages and in combination to animals (rats) with fibrosarcoma, metastatic tumor of the connective tissues, induced. The anticancer drug (20 mg/kg body weight) and the vitamin-E (400 mg/kg body weight) was administered for a period of 28 days from the day of tumor transplantation. The individual and the combined effects of these two substances were investigated by checking the growth of the tumor. Tumor markers like lactate dehydrogenase (LDH), serum glutamate pyruvate transminase (SGPT), serum glutamate oxaloacetate transaminase (SGOT), acid phosphatase, and alkaline phosphatase were analyzed for the changes in their concentration in serum, liver, and kidney to assess the success of the therapy. The increased level of the enzymes in the fibrosarcoma-suffering rats (GPII) was reduced by cyclophosphamide treatment (GP III) and vitamin E administration (GP IV). Among the treated groups, the combination therapy (GP V) showed greater efficacy in the treatment of fibrosarcoma than did individual administration, as there was more reduction in the levels of enzymes in Group V than those in to Groups III and IV. The enzyme levels were brought to near the normal level.


Molecular clutch drives cell response to surface viscosity.
Cell response to matrix rigidity has been explained by the mechanical properties of the actin-talin-integrin-fibronectin clutch. Here the molecular clutch model is extended to account for cell interactions with purely viscous surfaces (i.e., without an elastic component). Supported lipid bilayers present an idealized and controllable system through which to study this concept. Using lipids of different diffusion coefficients, the mobility (i.e., surface viscosity) of the presented ligands (in this case RGD) was altered by an order of magnitude. Cell size and cytoskeletal organization were proportional to viscosity. Furthermore, there was a higher number of focal adhesions and a higher phosphorylation of FAK on less-mobile (more-viscous) surfaces. Actin retrograde flow, an indicator of the force exerted on surfaces, was also seen to be faster on more mobile surfaces. This has consequential effects on downstream molecules; the mechanosensitive YAP protein localized to the nucleus more on less-mobile (more-viscous) surfaces and differentiation of myoblast cells was enhanced on higher viscosity. This behavior was explained within the framework of the molecular clutch model, with lower viscosity leading to a low force loading rate, preventing the exposure of mechanosensitive proteins, and with a higher viscosity causing a higher force loading rate exposing these sites, activating downstream pathways. Consequently, the understanding of how viscosity (regardless of matrix stiffness) influences cell response adds a further tool to engineer materials that control cell behavior.


Modeling the Optical Constants of Diamonds from 0.06 to 30 eV.
The optical properties of diamonds are modeled over a wide spectral range with the modified Adachi's model. Model parameters were estimated by use of the acceptance-probability-controlled simulated annealing algorithm. The employed model is quite flexible, as it uses an adjustable broadening function at each critical point. The broadening function can vary over a range of functions with similar kernels but different wings, so that extended absorption tails inherent to the conventional Lorentzian broadening can be eliminated. Good agreement with the experimental data is obtained in the entire investigated range. The obtained relative rms error for the real part of the index of refraction equals 4.7%, whereas for the imaginary part of the index-of-refraction relative rms error is 3.6%.


Effect of IL-4 on peripheral blood lymphocyte proliferation: implication in immunopathogenesis of H. pylori infection.
Interleukin (IL) 4 is a type 2 cytokine which has a negative immunoregulatory role in human infection. IL-4 suppresses the production of interferon-gamma and enhances IL-10 synthesis. However, the effect of IL-4 on proliferative response of lymphocytes remains to be elucidated. We have previously reported an increase in production of IL-4 in subjects with Helicobacter pylori (H. pylori) infection. To evaluate whether the increased IL-4 is responsible for the down-regulation of immune responses in H. pylori infection, we observed the proliferative response of peripheral blood lymphocytes (PBL) co-cultured with phythaemagglutinin (PHA) or H. pylori in the presence and absence of added IL-4. As we have previously shown, PHA and H. pylori may increase PBL proliferation (P < 0.001). An increase in PBL proliferation was observed when PBL were co-cultured with PHA (P < 0.001) or H. pylori (P < 0.001) in the presence of IL-4 compared to that in the absence of IL-4. The optimal dose of IL-4 to give maximal lymphocyte proliferation is 50 pg/ml for the PHA-stimulated group or 100 pg/ml for the H. pylori-stimulated group. The data suggest that the increased IL-4 does not directly contribute to suppressed lymphocyte proliferation in H. pylori infection. Further studies will be required to determine the role of IL-4 in other aspects of down-regulation of immune responses in H. pylori infection.


Cerebral palsy and growth failure at 6 to 7 years.
To evaluate the association between severity of cerebral palsy (CP) and growth to 6 to 7 years of age among children with moderate to severe (Mod/Sev) hypoxic ischemic encephalopathy (HIE). It was hypothesized that children with Mod/Sev CP would have poorer growth, lower cognitive scores, and increased rehospitalization rates compared with children with no CP (No CP). Among 115 of 122 surviving children followed in the hypothermia trial for neonatal HIE, growth parameters and neurodevelopmental status at 18 to 22 months and 6 to 7 years were available. Group comparisons (Mod/Sev CP and No CP) with unadjusted and adjusted analyses for growth <10th percentile and z scores by using Fisher's exact tests and regression modeling were conducted. Children with Mod/Sev CP had high rates of slow growth and cognitive and motor impairment and rehospitalizations at 18 to 22 months and 6 to 7 years. At 6 to 7 years of age, children with Mod/Sev CP had increased rates of growth parameters <10th percentile compared with those with No CP (weight, 57% vs 3%; height, 70% vs 2%; and head circumference, 82% vs 13%; P < .0001). Increasing severity of slow growth was associated with increasing age (P < .04 for weight, P < .001 for length, and P < .0001 for head circumference). Gastrostomy feeds were associated with better growth. Term children with HIE who develop Mod/Sev CP have high and increasing rates of growth <10th percentile by 6 to 7 years of age. These findings support the need for close medical and nutrition management of children with HIE who develop CP.


Cellular pharmacokinetic and pharmacodynamic analyses of ethacrynic acid: Implications in topical drug delivery in the eye.
Ethacrynic acid (ECA) is a potential trabecular meshwork (TM) drug that has shown promising results in preclinical studies for treatment of primary open-angle glaucoma. However, topical application of ECA is currently limited by adverse effects in corneal tissues. To this end, we developed a new theoretical model to evaluate time-dependent toxicity induced by ECA in corneal epithelial cells. The model consisted of a cellular pharmacokinetic (PK) module to determine intracellular concentration of ECA, and a pharmacodynamic (PD) module to determine the cytotoxicity of ECA. It was assumed that ECA-induced cytotoxicity depended on drug exposure time and peak concentration of bound ECA in cells. In addition to the model development, we experimentally determined the intracellular concentration of ECA as a function of drug dose and treatment time. The intracellular concentration increased linearly (i.e., no saturation) with increasing the dose of ECA. It also increased initially with time and then reached a steady-state at ~40 min. The percent of cells survived after treatment decreased with increasing the dose of drug or the time of treatment. The experimental data were fit by the new PK and PD models to obtain values of model constants. One of the unique applications of these models was to predict cell survival relative to control when extracellular concentration of ECA varied with time. The prediction showed that the toxicity of ECA might be significantly overestimated by using the traditional LC(50) determined in vitro. The new PK and PD models developed in this study were capable to fit experimental data and predict time-dependent toxicity of ECA in corneal epithelial cells. The models may be useful for optimizing the dose and schedule in topical application of ECA for glaucoma treatment.


Cloning, expression, and tissue distribution of the rat homolog of the bovine alpha 1C-adrenergic receptor provide evidence for its classification as the alpha 1A subtype.
Three alpha 1-adrenergic receptors (ARs) have been cloned, i.e., the alpha 1B-, alpha 1C-, and alpha 1D-ARs. Compared with the alpha 1B subtype, the alpha 1A subtype in tissue is described as being insensitive to chloroethylclonidine and sensitive to SZL-49 and having a 10-100-fold higher affinity for a number of agonists and antagonists. The alpha 1A subtype is also expressed in a variety of rat tissues (as assessed by pharmacology), with greatest abundance in the cerebral cortex, hippocampus, vas deferens, and submaxillary gland. The cloned bovine alpha 1C-AR, though having an alpha 1A-AR pharmacology, was first reported as not being expressed in any rat tissue (as determined by Northern analysis) and was therefore designated as a new subtype. We report the cloning, expression, and characterization of the rat homolog of the bovine alpha 1C-AR. Using a human alpha 1C-AR probe obtained by polymerase chain reaction screening of a neuroblastoma cell line (SK-N-MC), both exon 1 and exon 2 of the rat alpha 1C-AR gene were cloned from a rat genomic library. These two exons were spliced together and cloned into the expression vector pMT2'. Transfection into COS-1 cells and analysis of the ligand-binding profile of the expressed protein receptor using 125I-HEAT revealed a 10-100-fold higher affinity for the alpha 1-AR antagonists 5-methylurapidil, (+)-niguldipine, WB-4101, and phentolamine and the agonists oxymetazoline and methoxamine, compared with the alpha 1B-AR. This ligand-binding profile is similar to that for endogenously expressed tissue alpha 1A-ARs. In addition, the rat alpha 1C-AR was the least sensitive of the three cloned subtypes to the alkylating effects of chloroethylclonidine but was the most sensitive to the alkylating prazosin analog SZL-49, properties also observed for the tissue alpha 1A subtype. Furthermore, by three different techniques, i.e., RNase protection assays, reverse transcription-polymerase chain reaction Northern blotting, and in situ hybridization histochemistry, the rat alpha 1C-AR mRNA was localized to alpha 1A-AR-rich tissues, such as rat vas deferens, hippocampus, aorta, and submaxillary gland. Taken together, these data suggest that this receptor may actually represent the alpha 1A subtype.


The beneficial effects of nutraceuticals and natural products on small dense LDL levels, LDL particle number and LDL particle size: a clinical review.
Cardiovascular diseases (CVDs) are globally the major causes of morbidity and mortality. Evidence shows that smaller and denser low-dense lipoprotein (sdLDL) particles are independent atherogenic risk factors for CVD due to their greater susceptibility to oxidation, and permeability in the endothelium of arterial walls. sdLDL levels are an independent risk factor and of more predictive value than total LDL-C for the assessment of coronary artery disease and metabolic syndrome. Functional food ingredients have attracted significant attention for the management of dyslipidemia and subsequently increase cardio-metabolic health. However, to date there is no study that has investigated the effect of these bioactive natural compounds on sdLDL levels. Therefore, the aim of the present review is to summarize the evidence accrued on the effect of special dietary ingredients such as omega-3 polyunsaturated fatty acids, nutraceuticals and herbal medicines on the levels of sdLDL, LDL particle number, and LDL particle size. Based on the results of the existing clinical trials this review suggests that natural products such as medicinal plants, nutraceuticals and omega-3 fatty acids can be used as adjunct or complementary therapeutic agents to reduce sdLDL levels, LDL particle numbers or increase LDL particle size and subsequently may prevent and treat CVD, with the advantage that theses natural agents are generally safe, accessible, and inexpensive.


Access to, use of and satisfaction with health services among adults enrolled in Brazil's Family Health Strategy: evidence from the 2008 National Household Survey.
To assess the effects of participation in Brazil's primary healthcare programme (the Family Health Strategy or FHS) on access, use and satisfaction with health services among adults. Data are from the 2008 National Household Survey (PNAD) on 264 754 adults. This cross-sectional analysis compares FHS enrollees to both non-enrollees and those with private health plans. We calculated predicted probabilities of each outcome stratified by household wealth quintile, rural/urban location and sex using robust Poisson regression. We performed propensity score analysis to assess the differences in access among FHS enrollees and the rest of the population, once relevant socio-demographic characteristics and other determinants of access were balanced. Compared to families with neither FHS enrolment nor private health plans, adult FHS enrollees were generally more likely to have a usual source of care, to have visited a doctor or dentist in the past 12 months, to have access to needed medications and to be satisfied with the care they received. The FHS effect was largest among urban dwellers and the poorest. The FHS appears to be associated with enhanced access to and utilization of health services in Brazil. However, it has not yet been able to match levels of access experienced by those with private health plans, perhaps because the population served by the FHS is among the poorest, most rural and least healthy in the country.


Effect of Re-acidification on Buffalo Grass Rhizosphere and Bulk Microbial Communities During Phytostabilization of Metalliferous Mine Tailings.
Phytostabilized highly acidic, pyritic mine tailings are susceptible to re-acidification over time despite initial addition of neutralizing amendments. Studies examining plant-associated microbial dynamics during re-acidification of phytostabilized regions are sparse. To address this, we characterized the rhizosphere and bulk bacterial communities of buffalo grass used in the phytostabilization of metalliferous, pyritic mine tailings undergoing re-acidification at the Iron King Mine and Humboldt Smelter Superfund Site in Dewey-Humboldt, AZ. Plant-associated substrates representing a broad pH range (2.35-7.76) were sampled to (1) compare the microbial diversity and community composition of rhizosphere and bulk compartments across a pH gradient, and (2) characterize how re-acidification affects the abundance and activity of the most abundant plant growth-promoting bacteria (PGPB; including N2-fixing) versus acid-generating bacteria (AGB; including Fe-cycling/S-oxidizing). Results indicated that a shift in microbial diversity and community composition occurred at around pH 4. At higher pH (>4) the species richness and community composition of the rhizosphere and bulk compartments were similar, and PGPB, such as Pseudomonas, Arthrobacter, Devosia, Phyllobacterium, Sinorhizobium, and Hyphomicrobium, were present and active in both compartments with minimal presence of AGB. In comparison, at lower pH (<4) the rhizosphere had a significantly higher number of species than the bulk (p < 0.05) and the compartments had significantly different community composition (unweighted UniFrac; PERMANOVA, p < 0.05). Whereas some PGPB persisted in the rhizosphere at lower pH, including Arthrobacter and Devosia, they were absent from the bulk. Meanwhile, AGB dominated in both compartments; the most abundant were the Fe-oxidizer Leptospirillum and Fe-reducers Acidibacter and Acidiphilium, and the most active was the Fe-reducer Aciditerrimonas. This predominance of AGB at lower pH, and even their minimal presence at higher pH, contributes to acidifying conditions and poses a significant threat to sustainable plant establishment. These findings have implications for phytostabilization field site management and suggest re-application of compost or an alternate buffering material may be required in regions susceptible to re-acidification to maintain a beneficial bacterial community conducive to long-term plant establishment.


[Fabrication of sensor for reactive oxygen species using gold electrodes modified with electropolymerized porphyrins and application for detection of stress of plants].
Reactive Oxygen Spiecies (ROS) such as superoxide anion radical (.O(2)(-)) act as signals for the activation of stress-response and defense pathways. However, excess ROS generated by perturbing .O(2)(-) homeostasis stimulated many environmental stress, including intense light, drought, temperature stress, herbicides, induce high radical toxicity. Consequently, quantitative analysis of .O(2)(-) is a subject of intense research, since most of ROS are derived from .O(2)(-). Iron meso-tetrakis(3-thienyl)porphyrin complexes were electropolymerized onto a Au wire electrode. The modified Au electrode were applied to .O(2)(-) sensor to detect catalytic oxidation current of .O(2)(-) which was generated as an intermediate during the oxidation of xanthine by catalystic XOD. It was revealed that the sensor was quantitative to measure .O(2)(-). The modified Au electrode were applied to measure oxidation current of .O(2)(-) in mung beans under environmental stress condition. Plants were grown in atmosphere, 25 degrees C and in black darkness. The other plants were exposed to oxygen excess. The oxidation current of .O(2)(-) were increased plants were grown by high-oxygen environment compared to plants were grown at atmosphere. This experiment was indicated that environmental stress such as hyperoxia induced excess .O(2)(-) and Au wire sensor using iron porphyrin complexes is capable of .O(2)(-) detection in plants under environmental stresses.


Occupational Stress Experienced by Nurses Working in a Greek Regional Hospital: A Cross-sectional Study.
Occupational stress is both psychologically and physically challenging and may lead to high rates of absenteeism, burnout, and turnover. Nursing is considered as one of the most stressful and demanding professions. This study aims to measure perceived stress levels among nurses in a Greek public general hospital. A cross-sectional study was conducted. A total of 157 nurses and nursing assistants took part. A self-administered questionnaire including sociodemographics and Expanded Nursing Stress Scale (ENSS) for stress assessment was used. Analysis was performed using Statistical Package for the Social Sciences version 20.0. Total stress mean(SD) score ranged in medium levels [136.27 (47.80)]. The most stressful situations were dealing with death and dying [18.29 (5.59)]; patients' and their families' demands [20.90 (7.12)], and uncertainty concerning treatment [22.19 (9.07)]. Discrimination [4.40 (4.25)] and conflicts with peers [12.07 (5.10)] provoked less stress. There was a statistically significant relationship between age and total stress (F = 4.23, p < 0.001) and all distinct stressors. Nurses between 30 and 34 years expressed higher stress in all cases except patients' and their families' demands stressor. Those nurses who were divorced expressed higher stress in death and dying subscale (F = 2.93, p = 0.035). Nursing assistants expressing higher stress as far as workload (t = -2.40, p = 0.017), conflicts with physicians (t = -2.19, p = 0.033), and problems with peers (t = -2.63, p = 0.009) were concerned, compared to nurses. The findings of the study are in line with other researchers' findings concerning stressors among nursing personnel in Greece. Appropriate measures for the prevention and management of specific stressors must therefore be taken.


Regulation of ciliary function by fibroblast growth factor signaling identifies FGFR3-related disorders achondroplasia and thanatophoric dysplasia as ciliopathies.
Cilia project from almost every cell integrating extracellular cues with signaling pathways. Constitutive activation of FGFR3 signaling produces the skeletal disorders achondroplasia (ACH) and thanatophoric dysplasia (TD), but many of the molecular mechanisms underlying these phenotypes remain unresolved. Here, we report in vivo evidence for significantly shortened primary cilia in ACH and TD cartilage growth plates. Using in vivo and in vitro methodologies, our data demonstrate that transient versus sustained activation of FGF signaling correlated with different cilia consequences. Transient FGF pathway activation elongated cilia, while sustained activity shortened cilia. FGF signaling extended primary cilia via ERK MAP kinase and mTORC2 signaling, but not through mTORC1. Employing a GFP-tagged IFT20 construct to measure intraflagellar (IFT) speed in cilia, we showed that FGF signaling affected IFT velocities, as well as modulating cilia-based Hedgehog signaling. Our data integrate primary cilia into canonical FGF signal transduction and uncover a FGF-cilia pathway that needs consideration when elucidating the mechanisms of physiological and pathological FGFR function, or in the development of FGFR therapeutics.


Oxidative stress of Burkholderia cenocepacia induces insertion sequence-mediated genomic rearrangements that interfere with macrorestriction-based genotyping.
Burkholderia cenocepacia can cause serious infections and epidemics in patients with cystic fibrosis (CF). A CF population in the Czech Republic experienced an epidemic outbreak caused by a B. cenocepacia ST-32 strain. The clonality of the isolates was evident by multilocus sequence typing; however, fingerprinting profiles obtained by pulsed-field gel electrophoresis (PFGE) showed substantial band variability. We investigated whether the PFGE pattern diversity resulted from genomic rearrangements mediated by insertion sequences (IS); in addition, we determined whether stressful growth conditions altered the transposition activity of these IS. DNA probes for IS commonly found in B. cenocepacia were designed using the B. cenocepacia J2315 genome. Southern hybridization analysis of ST-32 isolates demonstrated diversity in both the copy number and the insertion site for a homologue of ISBcen20. Movement of the ISBcen20 homologue was detected when the ST-32 isolate CZ1238 was exposed to oxidative stress (growth in the presence of H(2)O(2)). PFGE analysis of CZ1238 derivatives exposed to oxidative stress demonstrated genomic rearrangements. Interestingly, when the closely related B. cenocepacia strain J2315 was exposed to oxidative stress, no movement of ISBcen20 was detected. Since frameshift mutations are present within the transposases of all copies of this IS in J2315, our data suggest that the transposase is inactive. In summary, we have demonstrated for the first time that IS movement can be mediated by oxidative stress and can lead to genomic rearrangements in the CF pathogen B. cenocepacia. These IS movements may alter the PFGE fingerprints of isolates that are clonal by other typing methods.


Nontarget Mass Spectrometry Reveals New Perfluoroalkyl Substances in Fish from the Yangtze River and Tangxun Lake, China.
Nontarget high-resolution mass spectrometry (Nt-HRMS) has been proven useful for the identification of unknown poly- and perfluoroalkyl substances (PFASs) in commercial products and water, but applications to biological samples are limited. China is the major PFAS-manufacturing nation; thus, here, we adapted our Nt-HRMS methods to fish collected from the Yangtze River and Tangxun Lake to discover potentially bioaccumulative PFASs in aquatic organisms destined for human consumption. In addition to traditional PFASs, over 330 other fluorinated analytes belonging to 10 classes of PFASs were detected among the pooled fish livers, including 6 sulfonate classes, 2 amine classes, 1 carboxylate class, and 1 N-heterocycle class. One class was detected in samples from both locations, 8 classes were detected exclusively in Tangxun Lake fish, and 1 class was detected exclusively in Yangtze River fish, 10 km downstream of a fluorochemical manufacturing site where we first reported these substances in wastewater 3 years ago. Overall, 4 of the PFAS classes (>165 analytes) are reported for the first time here. Wider monitoring and toxicological testing should be a priority for understanding the health risks posed to people and wildlife exposed to these substances.


Photosynthetic reaction center mimicry: low reorganization energy driven charge stabilization in self-assembled cofacial zinc phthalocyanine dimer-fullerene conjugate.
By employing well-defined self-assembly methods, a biomimetic bacterial photosynthetic reaction center complex has been constructed, and photoinduced electron transfer originating in this supramolecular donor-acceptor conjugate has been investigated. The biomimetic model of the bacterial "special pair" donor, a cofacial zinc phthalocyanine dimer, was formed via potassium ion induced dimerization of 4,5,4',5',4'', 5'',4''',5'''-zinc tetrakis(1,4,7,10,13-pentaoxatridecamethylene)phthalocyanine. The dimer was subsequently self-assembled with functionalized fullerenes via "two-point" binding involving axial coordination and crown ether-alkyl ammonium cation complexation to form the donor-acceptor pair, mimicking the noncovalently bound entities of the bacterial photosynthetic reaction center. The adopted self-assembly methodology yielded a supramolecular complex of higher stability with defined geometry and orientation as revealed by the binding constant and computational optimized structure. Unlike the previously reported porphyrin analog, the present phthalocyanine macrocycle based model system exhibited superior electron-transfer properties including formation of a long-lived charge-separated state, a key step of the photosynthetic light energy conversion process. Detailed analysis of the kinetic data in light of the Marcus theory of electron transfer revealed that small reorganization energy of the relatively rigid phthalocyanine is primarily responsible for slower charge-recombination process. The importance of the cofacial dimer in stabilizing the charge-separated state is borne out in the present all-supramolecular "reaction center" donor-acceptor mimic.


NMR structure of the second intracellular loop of the alpha 2A adrenergic receptor: evidence for a novel cytoplasmic helix.
A major, unresolved question in signal transduction by G protein coupled receptors (GPCRs) is to understand how, at atomic resolution, a GPCR activates a G protein. A step toward answering this question was made with the determination of the high-resolution structure of rhodopsin; we now know the intramolecular interactions that characterize the resting conformation of a GPCR. To what degree does this structure represent a structural paradigm for other GPCRs, especially at the cytoplasmic surface where GPCR-G protein interaction occurs and where the sequence homology is low among GPCRs? To address this question, we performed NMR studies on approximately 35-residue-long peptides including the critical second intracellular loop (i2) of the alpha 2A adrenergic receptor (AR) and of rhodopsin. To stabilize the secondary structure of the peptide termini, 4-12 residues from the adjacent transmembrane helices were included and structures determined in dodecylphosphocholine micelles. We also characterized the effects on an alpha 2A AR peptide of a D130I mutation in the conserved DRY motif. Our results show that in contrast to the L-shaped loop in the i2 of rhodopsin, the i2 of the alpha 2A AR is predominantly helical, supporting the hypothesis that there is structural diversity within GPCR intracellular loops. The D130I mutation subtly modulates the helical structure. The spacing of nonpolar residues in i2 with helical periodicity is a predictor of helical versus loop structure. These data should lead to more accurate models of the intracellular surface of GPCRs and of receptor-mediated G protein activation.


Kinetic consequences of the removal of a disulfide bridge on the folding of hen lysozyme.
Quenched-flow hydrogen exchange labeling, monitored by 1H NMR and electrospray ionization mass spectrometry (ESI-MS), has been employed in conjunction with stopped-flow circular dichroism and fluorescence to study the kinetic refolding from guanidinium chloride of a derivative of hen lysozyme in which one of the four disulfide linkages (Cys6-Cys127) has been selectively chemically reduced and carboxymethylated (CM6,127-lysozyme). Removal of this disulfide bridge has little effect on the structure and activity of the native enzyme, and the overall kinetics of refolding are very similar to those of the unmodified protein. A substantial amount of secondary structure is formed within 2 ms of the initiation of folding, followed by the slower formation of tertiary interactions characteristic of the native state, which are attained with a time constant (tau) of ca. 200 ms. There is clear evidence for fast and slow refolding populations, as in the intact protein. Folding of the three-disulfide derivative does, however, exhibit a major difference from that of the intact protein under the same final refolding conditions, in that the transient intermediate on the major refolding pathway of the intact protein, having persistent structure in the alpha-helical domain of the protein, is not detected by hydrogen exchange labeling during folding of the three-disulfide derivative. This suggests that the disulfide bond linking the N- and C-terminal regions of the protein is crucial for stabilization of the partially folded intermediate. In addition, the overshoot in the far-UV CD and the fluorescence minimum, both of which are attributed to non-native interactions, is not observed in the folding of CM6,127-lysozyme. That the lack of a detectable stable intermediate in the folding of CM6,127-lysozyme does not significantly affect the rate of attainment of the native state of the protein supports the proposed independent nature of the two folding domains and, as the Cys6-Cys127 disulfide bond is located in the alpha-domain, indicates that the rate-limiting step in folding of the intact protein, as well as of the three-disulfide derivative, involves stabilization of the beta-domain. The role of disulfide bridges in the formation and maintenance of the three-dimensional fold of proteins and in facilitating the observation of marginally stable intermediate species is discussed.


Ion channels formed by amphipathic helical peptides. A molecular modelling study.
Channel forming peptides (CFPs) are amphipathic peptides, of length ca. 20 residues, which adopt an alpha-helical conformation in the presence of lipid bilayers and form ion channels with electrophysiological properties comparable to those of ion channel proteins. We have modelled CFP channels as bundles of parallel trans-bilayer helices surrounding a central ion-permeable pore. Ion-channel interactions have been explored via accessible surface area calculations, and via evaluation of changes in van der Waals and electrostatic energies as a K+ ion is translated along the length of the pore. Two CFPs have been modelled: (a) zervamicin-A1-16, a synthetic apolar peptaibol related to alamethicin, and (b) delta-toxin from Staphylococcus aureus. Both of these CFPs have previously been shown to form ion channels in planar lipid bilayers, and have been shown to have predominantly helical conformations. Zervamicin-A1-16 channels were modelled as bundles of 4 to 8 parallel helices. Two related helix bundle geometries were explored. K(+)-channel interactions have been shown to involve exposed backbone carbonyl oxygen atoms. delta-Toxin channels were modelled as bundles of 6 parallel helices. Residues Q3, D11 and D18 generate favourable K(+)-channel interactions. Rotation of W15 about its C beta-C gamma bond has been shown to be capable of occluding the central pore, and is discussed as a possible model for sidechain conformational changes in relation to ion channel gating.


Flow structures at the proximal side-to-end anastomosis. Influence of geometry and flow division.
Flow structures were visualized in transparent polyurethane models of proximal side-to-end vascular anastomoses, using planar illumination of suspended tracer particles. Both the effects of geometry and flow division were determined under steady and pulsatile flow conditions, for anastomosis angles of 15, 30, and 45 degrees. The flow patterns were highly three-dimensional and were characterized by a series of vortices in the fully occluded distal artery and two helical vortices aligned with the axis of the graft. In steady flow, above a critical Reynolds number, the flow changed from a laminar regime to one displaying time-dependent behavior. In particular, significant fluctuating velocity components were observed in the distal artery and particles were shed periodically from the occluded artery into the graft. Pairs of asymmetric flow patterns were also observed in the graft, before the onset of the time-dependent flow regime. The critical Reynolds number ranged from 427 to 473 and appeared to be independent of anastomosis angle. The presence of a patent distal artery had a significant effect on the overall flow pattern and led to the formation of a large recirculation region at the toe of the anastomosis. The main structures observed in steady flow, such as vortices in the distal artery and helical flow in the graft, were also seen during the pulsatile cycle. However, the secondary flow components in the graft were more pronounced in pulsatile flow particularly during deceleration of the flow waveform. At higher mean Reynolds numbers, there was also a greater mixing between fluid in the occluded arterial section and that in the graft.


Localization and activity of various lysosomal proteases in Leishmania amazonensis-infected macrophages.
In mammalian hosts, Leishmania amastigotes are obligatory intracellular parasites of macrophages and multiply within parasitophorous vacuoles of phagolysosomal origin. To understand how they escape the harmful strategies developed by macrophages to kill ingested microorganisms, it is important to obtain information on the functional state of parasitophorous vacuole. For this purpose, we studied the intracellular distribution and activity of host lysosomal proteases in rat bone marrow-derived macrophages infected with Leishmania amazonensis amastigotes. Localization of cathepsins B, H, L, and D was investigated by using specific immunoglobulins. In uninfected macrophages, these enzymes were located in perinuclear granules (most of them were probably secondary lysosomes) which, after infection, disappeared progressively. In infected macrophages, cathepsins were detected mainly in the parasitophorous vacuoles, suggesting that the missing secondary lysosomes had fused with these organelles. Biochemical assays of various proteases (cathepsins B, H, and D and dipeptidyl peptidases I and II) showed that infection was accompanied by a progressive increase of all activities tested, except that of dipeptidyl peptidase II, which remained constant. No more than 1 to 10% of these activities could be attributed to amastigotes. These data indicate that (i) Leishmania infection is followed by an increased synthesis and/or a reduced catabolism of host lysosomal proteases, and (ii) amastigotes grow in a compartment rich in apparently fully active proteases. Unexpectedly, it was found that infected and uninfected macrophages degraded endocytosed proteins similarly. The lack of correlation in infected macrophages between increase of protease activities and catabolism of exogenous proteins could be linked to the huge increase in volume of the lysosomal compartment.


Symptom trends in the last year of life from 1998 to 2010: a cohort study.
Calls for improvement in end-of-life care have focused attention on the management of pain and other troubling symptoms at the end of life. To describe changes in pain intensity and symptom prevalence during the last year of life from 1998 to 2010. Observational study. The HRS (Health and Retirement Study), a nationally representative longitudinal survey of community-dwelling U.S. residents aged 51 years or older. 7204 HRS participants who died while enrolled in the study and their family respondents. Proxy-reported pain during the last year of life and other symptoms for at least 1 month during the last year of life. Trends in pain intensity and symptom prevalence were analyzed for all decedents and within the categories of sudden death, cancer, congestive heart failure or chronic lung disease, and frailty. Between 1998 and 2010, proxy reports of the prevalence of any pain increased for all decedents from 54.3% (95% CI, 51.6% to 57.1%) to 60.8% (CI, 58.2% to 63.4%), an increase of 11.9% (CI, 3.1% to 21.4%). Reported prevalences of depression and periodic confusion also increased for all decedents by 26.6% (CI, 14.5% to 40.1%) and 31.3% (CI, 18.6% to 45.1%), respectively. Individual symptoms increased in prevalence among specific decedent categories, except in cancer, which showed no significant changes. The prevalence of moderate or severe pain did not change among all decedents or in any specific decedent category. Use of proxy reports and limited information about some patient and surrogate variables. Despite national efforts to improve end-of-life care, proxy reports of pain and other alarming symptoms in the last year of life increased from 1998 to 2010. National Institute of Nursing Research.


Shared Sanitation Management and the Role of Social Capital: Findings from an Urban Sanitation Intervention in Maputo, Mozambique.
Shared sanitation-sanitation facilities shared by multiple households-is increasingly common in rapidly growing urban areas in low-income countries. However, shared sanitation facilities are often poorly maintained, dissuading regular use and potentially increasing disease risk. In a series of focus group discussions and in-depth interviews, we explored the determinants of shared sanitation management within the context of a larger-scale health impact evaluation of an improved, shared sanitation facility in Maputo, Mozambique. We identified a range of formal management practices users developed to maintain shared sanitation facilities, and found that management strategies were associated with perceived latrine quality. However-even within an intervention context-many users reported that there was no formal system for management of sanitation facilities at the compound level. Social capital played a critical role in the success of both formal and informal management strategies, and low social capital was associated with collective action failure. Shared sanitation facilities should consider ways to support social capital within target communities and identify simple, replicable behavior change models that are not dependent on complex social processes.


Plasma lipids and lipoprotein cholesterol concentrations in people with different diets in Britain.
Concentrations of total cholesterol and cholesterol in the various lipoprotein fractions were measured in vegans, vegetarians, fish eaters (who did not eat meat), and meat eaters. Total and low density lipoprotein cholesterol concentrations were higher in meat eaters than vegans, with vegetarians and fish eaters having intermediate and similar values. High density lipoprotein cholesterol concentration was highest in the fish eaters but did not differ among the other groups. There were striking trends with age in total and low density lipoprotein cholesterol concentrations, which differed between men and women: women showed a steady increase in concentration with age, whereas concentrations in men did not increase appreciably after the age of 40, which may partly explain sex differences in the prevalence of coronary heart disease. The differences in total cholesterol concentration suggest that the incidence of coronary heart disease may be 24% lower in lifelong British vegetarians and 57% lower in lifelong vegans than in meat eaters.


Dynamics of driven translocation of semiflexible polymers.
We study translocation of semiflexible polymers driven by force f_{d} inside a nanometer-scale pore using our three-dimensional Langevin dynamics model. We show that the translocation time τ increases with increasing bending rigidity κ. Similarly, the exponent β for the scaling of τ with polymer length N,τ∼N^{β}, increases with increasing κ as well as with increasing f_{d}. By comparing waiting times between semiflexible and fully flexible polymers we show that for realistic f_{d} translocation dynamics is to a large extent, but not completely, determined by the polymer's elastic length measured in number of Kuhn segments N_{Kuhn}. Unlike in driven translocation of flexible polymers, friction related to the polymer segment on the trans side has a considerable effect on the resulting dynamics. This friction is intermittently reduced by buckling of the polymer segment in the vicinity of the pore opening on the trans side. We show that in the experimentally relevant regime where viscosity is higher than in computer simulation models, the probability for this buckling increases with increasing f_{d}, giving rise to a larger contribution to the trans side friction at small f_{d}. Similarly to flexible polymers, we find significant center-of-mass diffusion of the cis side polymer segment which speeds up translocation. This effect is larger for smaller f_{d}. However, this speedup is smaller than the slowing down due to the trans side friction. At large enough N_{Kuhn}, the roles can be seen to be reversed, and the dynamics of flexible polymers can be reached. However, for example, polymers used in translocation experiments of DNA are elastically so short that the finite-length dynamics outlined here applies.


Candidate gene association mapping of Arabidopsis flowering time.
The pathways responsible for flowering time in Arabidopsis thaliana comprise one of the best characterized genetic networks in plants. We harness this extensive molecular genetic knowledge to identify potential flowering time quantitative trait genes (QTGs) through candidate gene association mapping using 51 flowering time loci. We genotyped common single nucleotide polymorphisms (SNPs) at these genes in 275 A. thaliana accessions that were also phenotyped for flowering time and rosette leaf number in long and short days. Using structured association techniques, we find that haplotype-tagging SNPs in 27 flowering time genes show significant associations in various trait/environment combinations. After correction for multiple testing, between 2 and 10 genes remain significantly associated with flowering time, with CO arguably possessing the most promising associations. We also genotyped a subset of these flowering time gene SNPs in an independent recombinant inbred line population derived from the intercrossing of 19 accessions. Approximately one-third of significant polymorphisms that were associated with flowering time in the accessions and genotyped in the outbred population were replicated in both mapping populations, including SNPs at the CO, FLC, VIN3, PHYD, and GA1 loci, and coding region deletions at the FRI gene. We conservatively estimate that approximately 4-14% of known flowering time genes may harbor common alleles that contribute to natural variation in this life history trait.


Herpes simplex virus 1 alpha regulatory protein ICP0 functionally interacts with cellular transcription factor BMAL1.
The infected cell protein no. 0 (ICP0) of herpes simplex virus 1 (HSV-1) is a promiscuous transactivator shown to enhance the expression of gene introduced into cells by infection or transfection. At the molecular level, ICP0 is a 775-aa ring finger protein localized initially in the nucleus and late in infection in the cytoplasm and mediates the degradation of several proteins and stabilization of others. None of the known functions at the molecular level account for the apparent activity of ICP0 as a transactivator. Here we report that ICP0 functionally interacts with cellular transcription factor BMAL1, a member of the basic helix-loop-helix PER-ARNT-SIM (PAS) super family of transcriptional regulators. Specifically, sequences mapped to the exon II of ICP0 interacted with BMAL1 in the yeast two-hybrid system and in reciprocal pull-down experiments in vitro. Moreover, the enhancement of transcription of a luciferase reporter construct whose promoter contained multiple BMAL1-binding sites by ICP0 and BMAL1 was significantly greater than that observed by ICP0 or BMAL1 alone. Although the level of BMAL1 present in nuclei of infected cells remained unchanged between 3 and 8 h after infection, the level of cytoplasmic BMAL1 was reduced at 8 h after infection. The reduction of cytoplasmic BMAL1 was significantly greater in cells infected with the ICP0-null mutant than in the wild-type virus-infected cells, suggesting that ICP0 mediates partial stabilization of the protein. These results indicate that ICP0 interacts physically and functionally with at least one cellular transcription-regulatory factor.


Changes in physical activity and other lifeway patterns influencing longevity.
We studied the adoption or maintenance of physical activity and other optional lifeway patterns for their influence on mortality rates of Harvard College alumni. Men aged 45-84 in 1977, surveyed by questionnaire in 1962 or 1966 and again in 1977, were followed from 1977 through 1988 or to age 90. Of 14,786 alumni, 2,343 died in 165,402 man-years of follow-up. Relative risks of death, standardized for potential confounding influences, for men who between questionnaires increased their physical activity through walking, stair climbing, and sports or recreational activities to 1,500 kcal or more per wk were 0.72 (95% confidence interval 0.64-0.82), compared with 1.00 for men who remained less active. Corresponding relative risks for men who adopted moderately vigorous sports play (> or = 4.5 METs) were 0.73 (0.65-0.81) vs 1.00 for men not adopting such sports; and for cigarette smokers who quit, 0.74 (0.65-0.84) vs 1.00 for persistent smokers. Men with recently diagnosed hypertension had a lower death risk than long-term hypertensives (0.80; 0.70-0.92), as did men with consistent normotension (0.52; 0.47-0.58). Changes in body-mass index had little influence on mortality during follow-up. These findings fit the hypothesis that adopting a physically active lifeway, quitting cigarette smoking, and remaining normotensive independently delay all-cause mortality and extend longevity.


Functional characterization of 3D protein structures informed by human genetic diversity.
Sequence variation data of the human proteome can be used to analyze 3D protein structures to derive functional insights. We used genetic variant data from nearly 140,000 individuals to analyze 3D positional conservation in 4,715 proteins and 3,951 homology models using 860,292 missense and 465,886 synonymous variants. Sixty percent of protein structures harbor at least one intolerant 3D site as defined by significant depletion of observed over expected missense variation. Structural intolerance data correlated with deep mutational scanning functional readouts for PPARG, MAPK1/ERK2, UBE2I, SUMO1, PTEN, CALM1, CALM2, and TPK1 and with shallow mutagenesis data for 1,026 proteins. The 3D structural intolerance analysis revealed different features for ligand binding pockets and orthosteric and allosteric sites. Large-scale data on human genetic variation support a definition of functional 3D sites proteome-wide.


Microfluidic channel-coupled 3D quartz nanohole arrays for high capture and release efficiency of BT20 cancer cells.
Nanostructured materials, such as silicon nanowires, quartz nanostructures, and polymer-modified nanostructures, are a promising new class of materials for the capture and enumeration of very rare tumor cells, including circulating tumor cells (CTCs), to examine their biological characteristics in whole blood of cancer patients. These cells can then be used for transplantation, anti-tumor cell therapy, and cell-secreted protein studies. It is believed that 3-dimensional (3D) nanostructured substrates efficiently enhance cell capture yields due to the increased local contacts between the 3D nanostructures and extracellular extensions of the tumor cells. Recent studies have been performed with enhanced cell capture yields thanks to various nanostructured platforms; however, there remains an urgent need both to capture and release viable rare tumor cells for further molecular (i.e., protein) analysis and to develop patient-specific drugs. Here, we first demonstrate that our 3D quartz nanohole array (QNHA) tumor cell capture and release system allows us not only to selectively capture rare tumor cells, but also to release the cells with high capture and release rates. This system was developed using streptavidin (STR)-functionalized QNHA (STR-QNHA) with a microfluidic channel. Our system has ideal cell-separation yields of as high as 85-91% and high release rates of >90% for the BT20 cell line. We suggest that the use of a microfluidic channel technique coupled with a STR-QNHA cell capture and release chip (STR-QNHA cell chip) would be a powerful and simple process to evaluate the capture, enumeration, and release of CTCs from patient whole blood for studying further cell therapy and tumor-cell-secreted molecules.


A survey of mineral status of soil, feeds and cattle in the Selale Ethiopian highlands. II. Trace elements.
The trace element status of crossbred Friesian x zebu (Boran) and local zebu cattle in the Selale highlands of Ethiopia was evaluated during the rainy and dry seasons of 1989 and 1990 in terms of the trace element content of soils, feeds, blood plasma and faeces on 25 randomly selected farms. Liver samples from animals of local breeds from a slaughter house were collected during the rainy and dry seasons of 1990 and analysed. Soil Fe and Mn were found to be high. Iron contents of all feeds were extremely high and Mn contents were higher than the dietary requirement. A large proportion of feed samples were deficient in Cu and zinc. There were wide variations in the concentrations of these elements among the soil and feed samples. Blood plasma Fe (P < 0.001) and Cu (P < 0.01) values differed significantly between the years. Plasma samples collected during 1990 contained lower mean Cu than 1989. The effect of season was statistically significant (P < 0.001) for all the trace elements. More than half of the animals were found to have low plasma Cu and Zn during the rainy season. Due to lack of any correlations between soil, pasture and blood plasma it is suggested that analyses of soils and pasture are not reliable for assessing the Fe, Cu and Zn status of grazing cattle in the highlands. Analyses of liver and blood appear to provide better indices.


Saliva viral load is a dynamic unifying correlate of COVID-19 severity and mortality.
While several clinical and immunological parameters correlate with disease severity and mortality in SARS-CoV-2 infection, work remains in identifying unifying correlates of coronavirus disease 2019 (COVID-19) that can be used to guide clinical practice. Here, we examine saliva and nasopharyngeal (NP) viral load over time and correlate them with patient demographics, and cellular and immune profiling. We found that saliva viral load was significantly higher in those with COVID-19 risk factors; that it correlated with increasing levels of disease severity and showed a superior ability over nasopharyngeal viral load as a predictor of mortality over time (AUC=0.90). A comprehensive analysis of immune factors and cell subsets revealed strong predictors of high and low saliva viral load, which were associated with increased disease severity or better overall outcomes, respectively. Saliva viral load was positively associated with many known COVID-19 inflammatory markers such as IL-6, IL-18, IL-10, and CXCL10, as well as type 1 immune response cytokines. Higher saliva viral loads strongly correlated with the progressive depletion of platelets, lymphocytes, and effector T cell subsets including circulating follicular CD4 T cells (cTfh). Anti-spike (S) and anti-receptor binding domain (RBD) IgG levels were negatively correlated with saliva viral load showing a strong temporal association that could help distinguish severity and mortality in COVID-19. Finally, patients with fatal COVID-19 exhibited higher viral loads, which correlated with the depletion of cTfh cells, and lower production of anti-RBD and anti-S IgG levels. Together these results demonstrated that viral load - as measured by saliva but not nasopharyngeal - is a dynamic unifying correlate of disease presentation, severity, and mortality over time.


Injectable thermosensitive magnetic nanoemulsion hydrogel for multimodal-imaging-guided accurate thermoablative cancer therapy.
Ferrofluid-based magnetic hyperthermia of cancers has gained significant attention in recent years due to its excellent efficacy, few deleterious side effects and unlimited tissue penetration capacity. However, the high tumor osmotic pressure causes injection leakage and thus position imprecision because of the fluidity of the ferrofluid and the absence of multimodal imaging guidance, which create tremendous challenges for clinical application. Here, a body temperature-induced gelation strategy is constructed for accurate localized magnetic tumor regression based on the unique behaviors of a magnetic nanoemulsion hydrogel (MNH) within tumors. The rapid intra-tumor gelation can securely restrict the MNH in tumor tissue without diffusion and leakage. The magnetically induced nanoparticle assembly-enhanced heating in the hydrogel and the heat accumulation caused by crosslinking among the nanoemulsion droplets further increased the heating efficiency. Meanwhile, US/MR/NIR multimodal imaging can guide the whole therapeutic process, achieving excellent magnetic hyperthermia therapeutic efficiency. This work highlights the great promise for improving the magnetic hyperthermia efficiency and the precision of the injection site for localized tumor therapy.


Inverse but independent trends in obesity and fitness levels among Greek children: a time-series analysis from 1997 to 2007.
We examined secular trends in physical fitness and BMI status in 8- to 9-year-old Greek children during an 11-year period (1997-2007). Population data derived from a yearly health survey performed in over 85% of Greek schools. Anthropometric measurements and physical fitness tests from 651,582 children were analyzed. The gender- and age-specific BMI cut-off points by the International Obesity Task Force were used to define overweight/obesity. Aerobic performance decreased by 4.9% (p < 0.001) for boys and 4.4% (p < 0.001) for girls between 1997 and 2007 while obesity increased by approximately 50% in both genders (p < 0.001). Time-series analyses revealed that the increasing trends in obesity were independent of the reduction in fitness levels. An increase from 21% in 1997 to 48.2% in 2007 was observed in the prevalence of the low quartile of aerobic performance for girls (p < 0.001) and from 25.7% in 1997 to 38.7% in 2007 (p < 0.001) for boys. Approximately 80% and 85% of obese boys and girls, respectively, failed to pass the low quartile of all aerobic tests in 2007. Inverse but independent trends in obesity and fitness levels were observed among Greek children during an 11-year period (1997-2007), a fact that predisposes our children to serious health risks as they grow older.


Identification of SARS-CoV-2 3CL Protease Inhibitors by a Quantitative High-throughput Screening.
The outbreak of coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has emphasized the urgency to develop effective therapeutics. Drug repurposing screening is regarded as one of the most practical and rapid approaches for the discovery of such therapeutics. The 3C like protease (3CL pro ), or main protease (M pro ) of SARS-CoV-2 is a valid drug target as it is a specific viral enzyme and plays an essential role in viral replication. We performed a quantitative high throughput screening (qHTS) of 10,755 compounds consisting of approved and investigational drugs, and bioactive compounds using a SARS-CoV-2 3CL pro assay. Twenty-three small molecule inhibitors of SARS-CoV-2 3CL pro have been identified with IC50s ranging from 0.26 to 28.85 μM. Walrycin B (IC 50 = 0.26 µM), Hydroxocobalamin (IC 50 = 3.29 µM), Suramin sodium (IC 50 = 6.5 µM), Z-DEVD-FMK (IC 50 = 6.81 µM), LLL-12 (IC 50 = 9.84 µM), and Z-FA-FMK (IC 50 = 11.39 µM) are the most potent 3CL pro inhibitors. The activities of anti-SARS-CoV-2 viral infection was confirmed in 7 of 23 compounds using a SARS-CoV-2 cytopathic effect assay. The results demonstrated a set of SARS-CoV-2 3CL pro inhibitors that may have potential for further clinical evaluation as part of drug combination therapies to treating COVID-19 patients, and as starting points for chemistry optimization for new drug development.


Endoplasmic Reticulum Stress and Mitochondrial Function in Airway Smooth Muscle.
Inflammatory airway diseases such as asthma affect more than 300 million people world-wide. Inflammation triggers pathophysiology via such as tumor necrosis factor α (TNFα) and interleukins (e.g., IL-13). Hypercontraction of airway smooth muscle (ASM) and ASM cell proliferation are major contributors to the exaggerated airway narrowing that occurs during agonist stimulation. An emergent theme in this context is the role of inflammation-induced endoplasmic reticulum (ER) stress and altered mitochondrial function including an increase in the formation of reactive oxygen species (ROS). This may establish a vicious cycle as excess ROS generation leads to further ER stress. Yet, it is unclear whether inflammation-induced ROS is the major mechanism leading to ER stress or the consequence of ER stress. In various diseases, inflammation leads to an increase in mitochondrial fission (fragmentation), associated with reduced levels of mitochondrial fusion proteins, such as mitofusin 2 (Mfn2). Mitochondrial fragmentation may be a homeostatic response since it is generally coupled with mitochondrial biogenesis and mitochondrial volume density thereby reducing demand on individual mitochondrion. ER stress is triggered by the accumulation of unfolded proteins, which induces a homeostatic response to alter protein balance via effects on protein synthesis and degradation. In addition, the ER stress response promotes protein folding via increased expression of molecular chaperone proteins. Reduced Mfn2 and altered mitochondrial dynamics may not only be downstream to ER stress but also upstream such that a reduction in Mfn2 triggers further ER stress. In this review, we summarize the current understanding of the link between inflammation-induced ER stress and mitochondrial function and the role played in the pathophysiology of inflammatory airway diseases.


PulseNet International: Vision for the implementation of whole genome sequencing (WGS) for global food-borne disease surveillance.
PulseNet International is a global network dedicated to laboratory-based surveillance for food-borne diseases. The network comprises the national and regional laboratory networks of Africa, Asia Pacific, Canada, Europe, Latin America and the Caribbean, the Middle East, and the United States. The PulseNet International vision is the standardised use of whole genome sequencing (WGS) to identify and subtype food-borne bacterial pathogens worldwide, replacing traditional methods to strengthen preparedness and response, reduce global social and economic disease burden, and save lives. To meet the needs of real-time surveillance, the PulseNet International network will standardise subtyping via WGS using whole genome multilocus sequence typing (wgMLST), which delivers sufficiently high resolution and epidemiological concordance, plus unambiguous nomenclature for the purposes of surveillance. Standardised protocols, validation studies, quality control programmes, database and nomenclature development, and training should support the implementation and decentralisation of WGS. Ideally, WGS data collected for surveillance purposes should be publicly available, in real time where possible, respecting data protection policies. WGS data are suitable for surveillance and outbreak purposes and for answering scientific questions pertaining to source attribution, antimicrobial resistance, transmission patterns, and virulence, which will further enable the protection and improvement of public health with respect to food-borne disease.


Effects of Low Intensity Exercise Against Apoptosis and Oxidative Stress in Streptozotocin-induced Diabetic Rat Heart.
Background The aim of this study was to investigate the effects of low intensity exercise on heart of streptozotocin (STZ)-induced diabetic rats. Materials and Methods The rats were randomly divided into 3 experimental groups: A (control), B (diabetic untreated), and C (diabetic treated with low intensity exercise); each group contains 8 animals. B and C groups received STZ. Diabetes was induced in 2 groups by a single intraperitoneal (i.p) injection of STZ (40 mg/kg, freshly dissolved in 0,1 M citrate buffer, pH 4.2). 2 days after STZ treatment, diabetes in 2 experimental groups was confirmed by measuring blood glucose levels. Rats with blood glucose levels of 250 mg/dl or higher were considered to be diabetic. Animals in the exercise group were made to run the treadmill once a day for 4 consecutive weeks. Exercise started 3 days prior to STZ administration. Results After induction of diabetes, histological abnormalities were observed, including myofibrillar loss, vacuolization of cytoplasm and irregularity of myofibrils. These alterations were attenuated by low intensity exercise. Our data indicates a significant reduction of oxidative stress and apoptosis in cardiomyocytes after exercise. Treatment of diabetic animals with low intensity exercise, decreased the elevated tissue malondialdehyde (MDA) levels and increased the reduced activities of the enzymatic antioxidants superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) in cardiac tissue. Conclusion These findings suggest that low intensity exercise has a therapeutic protective effect in diabetes by decreasing oxidative stress and apoptosis, and by preservation of myocardial integrity.


A transition state analogue of 5'-methylthioadenosine phosphorylase induces apoptosis in head and neck cancers.
Methylthio-DADMe-immucillin-A (MT-DADMe-ImmA) is an 86-pm inhibitor of human 5'-methylthioadenosine phosphorylase (MTAP). The sole function of MTAP is to recycle 5'-methylthioadenosine (MTA) to S-adenosylmethionine. Treatment of cultured cells with MT-DADMe-ImmA and MTA inhibited MTAP, increased cellular MTA concentrations, decreased polyamines, and induced apoptosis in FaDu and Cal27, two head and neck squamous cell carcinoma cell lines. The same treatment did not induce apoptosis in normal human fibroblast cell lines (CRL2522 and GM02037) or in MCF7, a breast cancer cell line with an MTAP gene deletion. MT-DADMe-ImmA alone did not induce apoptosis in any cell line, implicating MTA as the active agent. Treatment of sensitive cells caused loss of mitochondrial inner membrane potential, G(2)/M arrest, activation of mitochondria-dependent caspases, and apoptosis. Changes in cellular polyamines and MTA levels occurred in both responsive and nonresponsive cells, suggesting cell-specific epigenetic effects. A survey of aberrant DNA methylation in genomic DNA using a microarray of 12,288 CpG island clones revealed decreased CpG island methylation in treated FaDu cells compared with untreated cells. FaDu tumors in a mouse xenograft model were treated with MT-DADMe-ImmA, resulting in tumor remission. The selective action of MT-DADMe-ImmA on head and neck squamous cell carcinoma cells suggests potential as an agent for treatment of cancers sensitive to reduced CpG island methylation.


Effects of Spirulina on the functions and redox status of auditory system in senescence-accelerated prone-8 mice.
To our knowledge, the effects of Spirulina platensis water extract (SP) on hearing function have not yet been reported. This study investigated the effects of SP on the function and redox status of the auditory system. Auditory brainstem responses and redox status were compared between two groups of 3-month-old senescence-accelerated prone-8 (SAMP8) mice: the control group was fed a normal diet, and the experimental group was fed a normal diet with oral supplementation of SP for 6 weeks. Compared with the control group, the experimental group had significantly lower hearing thresholds according to auditory brainstem responses measured using click sounds and 8-kHz tone burst sound stimulation at the end of this study. The experimental group had a shorter I-III interval of auditory brainstem responses with 16-kHz tone burst stimulation than the control group that was of borderline significance. Additionally, the experimental group had significantly higher mRNA expression of the superoxide dismutase and catalase genes in the cochlea and brainstem and significantly higher mRNA expression of the glutathione peroxidase gene in the cochlea. Further, the experimental group had significantly lower malondialdehyde levels in the cochlea and brainstem than the control group. However, tumor necrosis factor-α mRNA expression was not significantly different between the control and experimental groups. SP could decrease hearing degeneration in senescence-accelerated prone-8 mice possibly by increasing superoxide dismutase, catalase, and glutathione peroxidase gene expression and decreasing damage from oxidative stress in the cochlea and brainstem.


The upregulation of thiamine (vitamin B1) biosynthesis in Arabidopsis thaliana seedlings under salt and osmotic stress conditions is mediated by abscisic acid at the early stages of this stress response.
Recent reports suggest that vitamin B1 (thiamine) participates in the processes underlying plant adaptations to certain types of abiotic and biotic stress, mainly oxidative stress. Most of the genes coding for enzymes involved in thiamine biosynthesis in Arabidopsis thaliana have been identified. In our present study, we examined the expression of thiamine biosynthetic genes, of genes encoding thiamine diphosphate-dependent enzymes and the levels of thiamine compounds during the early (sensing) and late (adaptation) responses of Arabidopsis seedlings to oxidative, salinity and osmotic stress. The possible roles of plant hormones in the regulation of the thiamine contribution to stress responses were also explored. The expression of Arabidopsis genes involved in the thiamine diphosphate biosynthesis pathway, including that of THI1, THIC, TH1 and TPK, was analyzed for 48 h in seedlings subjected to NaCl or sorbitol treatment. These genes were found to be predominantly up-regulated in the early phase (2-6 h) of the stress response. The changes in these gene transcript levels were further found to correlate with increases in thiamine and its diphosphate ester content in seedlings, as well as with the enhancement of gene expression for enzymes which require thiamine diphosphate as a cofactor, mainly α-ketoglutarate dehydrogenase, pyruvate dehydrogenase and transketolase. In the case of the phytohormones including the salicylic, jasmonic and abscisic acids which are known to be involved in plant stress responses, only abscisic acid was found to significantly influence the expression of thiamine biosynthetic genes, the thiamine diphosphate levels, as well as the expression of genes coding for main thiamine diphosphate-dependent enzymes. Using Arabidopsis mutant plants defective in abscisic acid production, we demonstrate that this phytohormone is important in the regulation of THI1 and THIC gene expression during salt stress but that the regulatory mechanisms underlying the osmotic stress response are more complex. On the basis of the obtained results and earlier reported data, a general model is proposed for the involvement of the biosynthesis of thiamine compounds and thiamine diphosphate-dependent enzymes in abiotic stress sensing and adaptation processes in plants. A possible regulatory role of abscisic acid in the stress sensing phase is also suggested by these data.


The role of nutritional support in the physical and functional recovery of critically ill patients: a narrative review.
The lack of benefit from randomised controlled trials has resulted in significant controversy regarding the role of nutrition during critical illness in terms of long-term recovery and outcome. Although methodological caveats with a failure to adequately appreciate biological mechanisms may explain these disappointing results, it must be acknowledged that nutritional support during early critical illness, when considered alone, may have limited long-term functional impact.This narrative review focuses specifically on recent clinical trials and evaluates the impact of nutrition during critical illness on long-term physical and functional recovery.Specific focus on the trial design and methodological limitations has been considered in detail. Limitations include delivery of caloric and protein targets, patient heterogeneity, short duration of intervention, inappropriate clinical outcomes and a disregard for baseline nutritional status and nutritional intake in the post-ICU period.With survivorship at the forefront of critical care research, it is imperative that nutrition studies carefully consider biological mechanisms and trial design because these factors can strongly influence outcomes, in particular long-term physical and functional outcome. Failure to do so may lead to inconclusive clinical trials and consequent rejection of the potentially beneficial effects of nutrition interventions during critical illness.


Stress pathways to health inequalities: Embedding ACEs within social and behavioral contexts.
This study addresses whether adverse childhood experiences (ACEs) demonstrate disproportional prevalence across demographic- and health-affecting characteristics, offer significant explanation of adult health outcomes, and show patterned association with illness susceptibility early within and across adulthood when viewed in combination with income and psychosocial resources. Data were derived from a population-based state health survey using stratified random sampling of household adults (n=7,470): ages 18-99 (M=55), 59.9% females, and race/ethnicity, income and education levels representative of the region. We assessed ACEs by aggregating 8 adversity forms, 5 health behaviors and 3 psychosocial resources; and health outcomes (number of chronic conditions, subjective wellness). Disproportionality was evident in ACEs levels by demographics, adult SES, health behaviors, and psychosocial resources in expected directions. Stepped multiple regressions of health outcomes demonstrated significant betas and R2 change for each predictor block, revealing cumulative as well as unique explanatory utility. Early onset chronic illness was evident on the basis of ACEs levels. These illnesses were amplified for low income respondents. Prevalence was highest across adulthood for those also reporting low psychosocial assets. Findings offer novel insights as to the "long reach" of childhood adversity on health, conditioned by circumstances under which these effects may occur. Health resilience offered by health behaviors and psychosocial resources should shape thinking about preventive and remedial interventions by social work and allied professionals across a range of settings.


Pharmacokinetics of Perfluorobutane after Intra-Venous Bolus Injection of Sonazoid in Healthy Chinese Volunteers.
Sonazoid is an ultrasound contrast agent based on microbubbles (MB) containing perfluorobutane (PFB) gas. Sonazoid is approved in Japan, Korea and Norway for contrast-enhanced ultrasonography of focal liver lesions and focal breast lesions (Japan only). The objective of this study was to determine the pharmacokinetics (PKs) and safety of Sonazoid in Chinese healthy volunteers (HVs) and to evaluate the potential for ethnic differences in PKs between Chinese and Caucasian HVs. Sonazoid was administered as an intra-venous bolus injection at the clinical dose of 0.12 μL or 0.60 μL MB/kg body weight to two groups of eight Chinese HVs. Expired air and blood samples were collected and analyzed using a validated gas chromatographic tandem mass spectrometry method, and the main PK parameters were calculated. The highest PFB concentrations in blood were observed shortly after intra-venous administration of Sonazoid, and elimination of PFB was rapid. In the 0.12 μL MB/kg body weight cohort, PFB concentrations above the limit of quantification were observed for only 10 to 15 min post-injection. In the 0.60 μL MB/kg body weight cohort, PFB concentrations above the limit of quantification were observed for 60 min post-injection, and the shape of the elimination curve suggested a biphasic elimination profile. The maximum observed concentration (Cmax) values of PFB in blood were 2.3 ± 1.1 and 19.1 ± 9.2 ng/g for the 0.12 and 0.60 μL MB/kg body weight dose groups (mean ± standard deviation). Area under the curve values were 10.1 ± 2.7 and 90.1 ± 38.3 ng × min/g for the 0.12 and 0.60 μL MB/kg body weight dose groups. Cmax values of PFB in exhaled air were 0.35 ± 0.2 and 2.4 ± 0.7 ng/mL for the 0.12 and 0.60 μL MB/kg body weight dose groups. Assessment of laboratory parameters, vital signs, oxygen saturation and electrocardiograms revealed no changes indicative of a concern. The PK profile and safety data generated in the Chinese HVs were comparable to previous data for Caucasian HVs.


An Examination of Measurement Procedures and Characteristics of Baseline Outcome Data in Single-Case Research.
There has been growing interest in using statistical methods to analyze data and estimate effect size indices from studies that use single-case designs (SCDs), as a complement to traditional visual inspection methods. The validity of a statistical method rests on whether its assumptions are plausible representations of the process by which the data were collected, yet there is evidence that some assumptions-particularly regarding normality of error distributions-may be inappropriate for single-case data. To develop more appropriate modeling assumptions and statistical methods, researchers must attend to the features of real SCD data. In this study, we examine several features of SCDs with behavioral outcome measures in order to inform development of statistical methods. Drawing on a corpus of over 300 studies, including approximately 1,800 cases, from seven systematic reviews that cover a range of interventions and outcome constructs, we report the distribution of study designs, distribution of outcome measurement procedures, and features of baseline outcome data distributions for the most common types of measurements used in single-case research. We discuss implications for the development of more realistic assumptions regarding outcome distributions in SCD studies, as well as the design of Monte Carlo simulation studies evaluating the performance of statistical analysis techniques for SCD data.


Twitter, time and emotions.
The study of temporal trajectories of emotions shared in tweets has shown that both positive and negative emotions follow nonlinear circadian (24 h) and circaseptan (7-day) patterns. But to this point, such findings could be instrument-dependent as they rely exclusively on coding using the Linguistic Inquiry Word Count. Further, research has shown that self-referential content has higher relevance and meaning for individuals, compared with other types of content. Investigating the specificity of self-referential material in temporal patterns of emotional expression in tweets is of interest, but current research is based upon generic textual productions. The temporal variations of emotions shared in tweets through emojis have not been compared to textual analyses to date. This study hence focuses on several comparisons: (i) between Self-referencing tweets versus Other topic tweets, (ii) between coding of textual productions versus coding of emojis, and finally (iii) between coding of textual productions using different sentiment analysis tools (the Linguistic Inquiry and Word Count-LIWC; the Valence Aware Dictionary and sEntiment Reasoner-VADER and the Hu Liu sentiment lexicon-Hu Liu). In a collection of more than 7 million Self-referencing and close to 18 million Other topic content-coded tweets, we identified that (i) similarities and differences in terms of shape and amplitude can be observed in temporal trajectories of expressed emotions between Self-referring and Other topic tweets, (ii) that all tools feature significant circadian and circaseptan patterns in both datasets but not always, and there is often a correspondence in the shape of circadian and circaseptan patterns, and finally (iii) that circadian and circaseptan patterns obtained from the coding of emotional expression in emojis sometimes depart from those of the textual analysis, indicating some complementarity in the use of both modes of expression. We discuss the implications of our findings from the perspective of the literature on emotions and well-being.


LC-MS/MS in the elucidation of an isomer of the recreational drug methylenedioxy ethylamphetamine: methylenedioxy dimethylamphetamine.
This paper describes the surplus value of a quadrupole-orthogonal acceleration TOF mass spectrometer, coupled to a liquid chromatographic separation system, for the unequivocal identification and structural elucidation of an unknown compound in the field of designer drugs. In a patient sample set (blood, tissues, vitreous humor, etc.), analyzed with a dedicated liquid chromatographic-fluorescence detection method for the determination of methylenedioxy amphetamine, methylenedioxy methamphetamine, and methylenedioxy ethylamphetamine (MDEA), a "strange" inexplicable peak appeared at a retention time not corresponding to any of our reference materials. Based on the identical excitation and emission wavelengths in detection, and a retention behavior comparable to MDEA, it was assumed that this unknown compound was an isomer of the recreational drug MDEA. With a simple and straightforward methodological crossover between LC fluorescence detection and LC-MS/MS, additional information for structural elucidation was easily obtained. Chromatographic separation was achieved on a Hypersil BDS C18 column (fluorescence detection part) and on a Hypersil BDS phenyl column (mass spectrometric detection part). MS showed that the unknown compound's molecular mass was identical to that of MDEA, and, in addition, its fragmentation pattern too proved quite similar to that of MDEA. A thorough literature overview and study of the fragmentation pattern by means of the MS/MS spectrum led to an evidence-based hypothesis of 3,4-methylenedioxy N,N-dimethylamphetamine (MDDM) being the unknown compound. To confirm this hypothesis, MDDM was synthesized and its presence in our biological sample was finally demonstrated by co-injection with alternatively synthesized MDDM and MDEA. This application shows the synergism between LC and MS in the elucidation of unknown compounds, nevertheless emphasizing the essence of chromatographic separation when dealing with isomers.


Cell seeding density is a critical determinant for copolymer scaffolds-induced bone regeneration.
Constructs intended for bone tissue engineering (TE) are influenced by the initial cell seeding density. Therefore, the objective of this study was to determine the effect of bone marrow stromal stem cells (BMSCs) density loaded onto copolymer scaffolds on bone regeneration. BMSCs were harvested from rat's bone marrow and cultured in media with or without osteogenic supplements. Cells were seeded onto poly(l-lactide-co-ε-caprolactone) [poly(LLA-co-CL)] scaffolds at two different densities: low density (1 × 10(6) cells/scaffold) or high density (2 × 10(6) cells/scaffold) using spinner modified flasks and examined after 1 and 3 weeks. Initial attachment and spread of BMSC onto the scaffolds was recorded by scanning electron microscopy. Cell proliferation was assessed by DNA quantification and cell differentiation by quantitative real-time reverse transcriptase-polymerized chain reaction analysis (qRT-PCR). Five-millimeter rat calvarial defects (24 defects in 12 rats) were implanted with scaffolds seeded with either low or high density expanded with or without osteogenic supplements. Osteogenic supplements significantly increased cell proliferation (p < 0.001). Scaffolds seeded at high cell density exhibited higher mRNA expressions of Runx2 p = 0.001, Col1 p = 0.001, BMP2 p < 0.001, BSP p < 0.001, and OC p = 0.013. More bone was formed in response to high cell seeding density (p = 0.023) and high seeding density with osteogenic medium (p = 0.038). Poly (LLA-co-CL) scaffolds could be appropriate candidates for bone TE. The optimal number of cells to be loaded onto scaffolds is critical for promoting Extracellular matrix synthesis and bone formation. Cell seeding density and osteogenic supplements may have a synergistic effect on the induction of new bone.


Iron Overload Exacerbates the Risk of Hemorrhagic Transformation After tPA (Tissue-Type Plasminogen Activator) Administration in Thromboembolic Stroke Mice.
Background and Purpose- Recanalization with tPA (tissue-type plasminogen activator) is the only pharmacological therapy available for patients with ischemic stroke. However, the percentage of patients who may receive this therapy is limited by the risk of hemorrhagic transformation (HT)-the main complication of ischemic stroke. Our aim is to establish whether iron overload affects HT risk, to identify mechanisms that could help to select patients and to prevent this devastating complication. Methods- Mice fed with control or high-iron diet were subjected to thromboembolic stroke, with or without tPA therapy at different times after occlusion. Blood samples were collected for determination of malondialdehyde, matrix metalloproteinases, and fibronectin. Brain samples were collected 24 hours after occlusion to determine brain infarct and edema size, hemorrhage extension, IgG extravasation, and inflammatory and oxidative markers (neutrophil infiltration, 4-hydroxynonenal, and matrix metalloproteinase-9 staining). Results- Despite an increased rate of recanalization, iron-overload mice showed less neuroprotection after tPA administration. Importantly, iron overload exacerbated the risk of HT after early tPA administration, accelerated ischemia-induced serum matrix metalloproteinase-9 increase, and enhanced basal serum lipid peroxidation. High iron increased brain lipid peroxidation at most times and neutrophil infiltration at the latest time studied. Conclusions- Our data showing that iron overload increases the death of the compromised tissues, accelerates the time of tPA-induced reperfusion, and exacerbates the risk of HT may have relevant clinical implications for a safer thrombolysis. Patients with stroke with iron overload might be at high risk of HT after fibrinolysis, and, therefore, clinical studies must be performed to confirm our results.


Social support and care needs of the disabled elderly population: An empirical study based on survey data from Beijing, China.
In order to describe and examine differences in social support and care needs among disabled Chinese elderly, the current study used stratified sampling to survey local residents of Beijing age 60 or over in the districts of Xicheng, Chaoyang, and Tongzhou in 2016. Structured in-person interviews were conducted with a 7-domain questionnaire. Multiple logistic regressions were used to compare social support and care needs among functioning, partially disabled, and completely disabled elderly. All statistical analyses were performed using SPSS 19.0 with a significance level of 0.05 (two-sided). One thousand and eighty-three residents completed the survey. Based on Activities of Daily Living (ADL) scores, 736 (68.0%) respondents were functioning (ADL score = 14), 167 (15.4%) were partially disabled (14 < ADL score < 22), and 180 (16.6%) were fully disabled (ADL score ≥ 22). Most of the disabled had formal financial support, they received daily care at home, and they received modest emotional support. After controlling for confounding factors, fully disabled respondents were 2.35 times (p = 0.018) more likely to receive financial support and 3.65 times (p = 0.003) more likely to receive emotional support than functioning respondents. However, the fully functioning and partially disabled did not differ significantly in terms of financial or emotional support. Compared to fully functioning respondents, partially disabled respondents were 0.49 (p < 0.001) times less likely to be fully satisfied with their daily care while fully disabled respondents were 0.37 (p < 0.001) times less likely to be fully satisfied with that care. The current study provided a thorough depiction of the current status of social support and care needs of disabled Chinese elderly. More attention should be paid to social support for the partially disabled and daily care for both the partially and fully disabled.