Does high-dose coenzyme Q10 improve oxidative damage and clinical outcomes in Parkinson's disease?
Evidence on the efficacy of high-dose coenzyme Q10 (CoQ10) in Parkinson's disease (PD) is conflicting. An open-label dose-escalation study was performed to examine the effects of CoQ10 on biomarkers of oxidative damage and clinical outcomes in 16 subjects with early idiopathic PD. Each dose (400, 800, 1200, and 2400 mg/day) was consumed daily for 2 weeks. High-dose CoQ10 was well tolerated and improvements in the total Unified Parkinson's Disease Rating Scale (median, 37 vs. 27; p=0.048) were observed following study completion. Plasma F2-isoprostanes (adjusted for arachidonate) were significantly reduced in the 400-1200 mg/day dose range, but increased at 2400 mg/day dosage. A similar pattern of change was observed with serum phospholipase A2 activities. Levels of plasma all trans-retinol, plasma total tocopherol, serum uric acid, and serum total cholesterol were unchanged despite an increase in the CoQ10 dosage. Subjects with symptomatic benefits from CoQ10 (decrease in total UPDRS >10 points) had lower baseline plasma ubiquinol (p=0.07, Mann-Whitney U test) and decreased F2-isoprostanes per unit arachidonate (p=0.04, Wilcoxon Signed-Ranks test). These results lead to the hypothesis that the therapeutic response to CoQ10 depends on baseline levels of ubiquinol and whether the dosage of CoQ10 used can ameliorate the burden of oxidative damage.


Target Flow-Pressure Operating Range for Designing a Failing Fontan Cavopulmonary Support Device.
Fontan operation as the current standard of care for the palliation of single ventricle defects results in significant late complications. Using a mechanical circulatory device for the right circulation to serve the function of the missing subpulmonary ventricle could potentially stabilize the failing Fontan circulation. This study aims to elucidate the hydraulic operating regions that should be targeted for designing cavopulmonary blood pumps. By integrating numerical analysis and available clinical information, the interaction of the cavopulmonary support via the IVC and full assist configurations with a wide range of simulated adult failing scenarios was investigated; with IVC and full assist corresponding to the inferior venous return or the entire venous return, respectively, being routed through the device. We identified the desired hydraulic operating regions for a cavopulmonary assist device by clustering all head pressures and corresponding pump flows that result in hemodynamic improvement for each simulated failing Fontan physiology. Results show that IVC support can produce beneficial hemodynamics in only a small fraction of failing Fontan scenarios. Cavopulmonary assist device could increase cardiac index by 35% and decrease the inferior vena cava pressure by 45% depending on the patient's pre-support hemodynamic state and surgical configuration of the cavopulmonary assist device (IVC or full support). The desired flow-pressure operating regions we identified can serve as the performance criteria for designing cavopulmonary assist devices as well as evaluating off-label use of commercially available left-side blood pumps for failing Fontan cavopulmonary support.


Organotypic brain explant culture as a drug evaluation system for malignant brain tumors.
Therapeutic options for malignant brain tumors are limited, with new drugs being continuously evaluated. Organotypic brain slice culture has been adopted for neuroscience studies as a system that preserves brain architecture, cellular function, and the vascular network. However, the suitability of brain explants for anticancer drug evaluation has been unclear. We here adopted a mouse model of malignant glioma based on expression of H-RasV12 in Ink4a/Arf-/- neural stem/progenitor cells to establish tumor-bearing brain explants from adult mice. We treated the slices with cisplatin, temozolomide, paclitaxel, or tranilast and investigated the minimal assays required to assess drug effects. Serial fluorescence-based tumor imaging was sufficient for evaluation of cisplatin, a drug with a pronounced cytotoxic action, whereas immunostaining of cleaved caspase 3 (a marker of apoptosis) and of Ki67 (a marker of cell proliferation) was necessary for the assessment of temozolomide action and immunostaining for phosphorylated histone H3 (a marker of mitosis) allowed visualization of paclitaxel-specific effects. Staining for cleaved caspase 3 was also informative in the assessment of drug toxicity for normal brain tissue. Incubation of explants with fluorescently labeled antibodies to CD31 allowed real-time imaging of the microvascular network and complemented time-lapse imaging of tumor cell invasion into surrounding tissue. Our results suggest that a combination of fluorescence imaging and immunohistological staining allows a unified assessment of the effects of various classes of drug on the survival, proliferation, and invasion of glioma cells, and that organotypic brain slice culture is therefore a useful tool for evaluation of antiglioma drugs.


Manganese oxide-modified biochar: production, characterization and applications for the removal of pollutants from aqueous environments - a review.
The development of manganese (Mn) oxides (MnOx) modified biochar (MnOBC) for the removal of pollutants from water has received significant attention. However, a comprehensive review focusing on the use of MnOBC for the removal of organic and inorganic pollutants from water is missing. Therefore, the preparation and characterization of MnOBC, and its capacity for the removal of inorganic (e.g., toxic elements) and organic (e.g., antibiotics and dyes) from water have been discussed in relation to feedstock properties, pyrolysis temperature, modification ratio, and environmental conditions here. The removal mechanisms of pollutants by MnOBC and the fate of the sorbed pollutants onto MnOBC have been reviewed. The impregnation of biochar with MnOx improved its surface morphology, functional group modification, and elemental composition, and thus increased its sorption capacity. This review establishes a comprehensive understanding of synthesizing and using MnOBC as an effective biosorbent for remediation of contaminated aqueous environments.


All-Arthroscopic Suture Fixation of Patellar Osteochondritis Dissecans.
Osteochondritis dissecans of the knee, despite its cause, is characterized by the impairment of the subchondral bone. Failure of its spontaneous healing makes surgical fixation often necessary. The patella is less affected than other locations in the knee. Its surgical treatment remains a challenge due to the thickness of the lesion and the complex approach of the retropatellar cartilage. Arthroscopy has the theoretical advantage to avoid a possible arthrotomy; however, the retrograde application of fixation materials does not guarantee good fragment compression and may lead to cartilage penetration and damage. The purpose of this Technical Note is to present a reproducible, full arthroscopic suture fixation technique for patellar osteochondritis dissecans lesions. By using the posterior cruciate tibial drill guide, absorbable sutures are passed through the center and the peripheral borders of the lesion resulting in a "spider-parachute-type" fixation with direct fragment compression.


Antecedents of geographical expansion: The case of federally qualified health centers.
Federally qualified health centers (FQHCs) are pivotal safety net primary care providers for the medically underserved. FQHCs have complex organizational designs, with many FQHCs providing care at multiple physical locations ("sites"). The number of sites, however, varies considerably between FQHCs, which can have important implications for differential access that may perpetuate disparities in quality of care. The objective of this study is to explore the organizational and environmental antecedents of the number of sites operated by each FQHC. The findings of this study contribute to a better understanding of FQHCs' expansion that has vital implications for cost and access outcomes. The study is based on data between the years 2012 and 2018. Using multivariate growth curve modeling, we analyzed the final sample, consisting of 5,482 FQHC-years. The level of competition, measured as the number of FQHC sites in the Primary Care Service Area (PCSA) and the number of primary care physicians per 1,000 PCSA residents, was positively associated with the number of FQHC sites. The number of patients, the level of federal grant, and the year were also positively associated with the number of FQHC sites, whereas percentage of Medicaid patients; workforce supply, measured as primary care physician assistants per 1,000 PCSA residents; Medicaid expansion; and state/local funding available for FQHCs were not. Findings of this study indicate that competition, especially between peer FQHCs, is significantly associated with FQHC expansion. This result suggests that FQHC managers and policymakers may closely monitor cost, access, and quality implications of competition and FQHC expansion.


Indomethacin increases the effect of isosorbide dinitrate on cerebral hemodynamic in migraine patients: pathogenetic and therapeutic implications.
Intracerebral vascular reactivity induced by the nitric oxide (NO) donor isosorbide dinitrate (IDN, 5 mg sublingually) is more major and longer-lasting in migraine patients who develop delayed headache in response to the drug. The headache is purportedly due to neuronally-mediated vascular mechanisms. Indomethacin inhibits prostaglandin synthesis, which is involved in NO generation. Indomethacin also decreases cerebral blood flow by constricting precapillary resistance vessels. In the present study, the hemodynamic effects of indomethacin were evaluated in migraine patients and healthy controls by means of transcranial Doppler monitoring. Indomethacin caused a significant decrease in mean flow velocity in the middle cerebral artery. This was an additional effect to the mean velocity decrease induced by IDN. The interactions between the two drugs suggest that their effects on cerebral hemodynamics (and pain) may be of relevance both in understanding the role of NO in migraine pathogenesis and in evaluating symptomatic treatments for migraine attacks.


Multi-scale mechanics of traumatic brain injury: predicting axonal strains from head loads.
The length scales involved in the development of diffuse axonal injury typically range from the head level (i.e., mechanical loading) to the cellular level. The parts of the brain that are vulnerable to this type of injury are mainly the brainstem and the corpus callosum, which are regions with highly anisotropically oriented axons. Within these parts, discrete axonal injuries occur mainly where the axons have to deviate from their main course due to the presence of an inclusion. The aim of this study is to predict axonal strains as a result of a mechanical load at the macroscopic head level. For this, a multi-scale finite element approach is adopted, in which a macro-level head model and a micro-level critical volume element are coupled. The results show that the axonal strains cannot be trivially correlated to the tissue strain without taking into account the axonal orientations, which indicates that the heterogeneities at the cellular level play an important role in brain injury and reliable predictions thereof. In addition to the multi-scale approach, it is shown that a novel anisotropic equivalent strain measure can be used to assess these micro-scale effects from head-level simulations only.


Phenotypic plasticity and adaptive evolution contribute to advancing flowering phenology in response to climate change.
Anthropogenic climate change has already altered the timing of major life-history transitions, such as the initiation of reproduction. Both phenotypic plasticity and adaptive evolution can underlie rapid phenological shifts in response to climate change, but their relative contributions are poorly understood. Here, we combine a continuous 38 year field survey with quantitative genetic field experiments to assess adaptation in the context of climate change. We focused on Boechera stricta (Brassicaeae), a mustard native to the US Rocky Mountains. Flowering phenology advanced significantly from 1973 to 2011, and was strongly associated with warmer temperatures and earlier snowmelt dates. Strong directional selection favoured earlier flowering in contemporary environments (2010-2011). Climate change could drive this directional selection, and promote even earlier flowering as temperatures continue to increase. Our quantitative genetic analyses predict a response to selection of 0.2 to 0.5 days acceleration in flowering per generation, which could account for more than 20 per cent of the phenological change observed in the long-term dataset. However, the strength of directional selection and the predicted evolutionary response are likely much greater now than even 30 years ago because of rapidly changing climatic conditions. We predict that adaptation will likely be necessary for long-term in situ persistence in the context of climate change.


Optimisation of cosolvent concentration for topical drug delivery III--influence of lipophilic vehicles on ibuprofen permeation.
Previously, we have reported the effects of water, ethanol, propylene glycol and various binary and ternary mixtures of these solvents on the permeation of ibuprofen in model membranes and in skin. The present study investigates the influence of lipophilic vehicles on the transport of ibuprofen in silicone membrane and in human skin. The permeation of ibuprofen was measured from mineral oil (MO), Miglyol® 812 (MG) and binary mixtures of MO and MG. The solubility of ibuprofen was 5-fold higher in MG than in MO, however, the permeation of ibuprofen from the pure vehicles and combinations of both was comparable in silicone membrane. Additionally, there were no significant differences in skin permeation for MO and MG vehicles. When the permeation of various hydrophilic and lipophilic vehicles is considered, a trend between flux values for the model membrane and skin is evident (r(2) = 0.71). The findings suggest that silicone membrane may provide information on qualitative trends in skin permeation for vehicles of diverse solubility and partition characteristics.


Conformational dynamics and thermal stabilities of the alphabeta-tubulin dimer: a molecular dynamics simulation study.
Molecular dynamics simulations were employed to study the conformational dynamics and thermal stabilities of the large alphabeta-tubulin dimer systems under various conditions. The alphabeta-tubulin dimer at ambient and cryogenic temperatures remains stable around the native conformations, and the water solvent molecules tend to stabilize the native conformations through the formation of complex hydrogen-bonding networks. The elevation of temperature severely destroys the native conformations and the bioactivities, especially in aqueous solution. It was caused by the lack of intra- and inter-molecular interactions and the decrease of water polarity at higher temperatures. The conformational analysis of the selected segment (beta15-QIGAKFWEVIS) provides detailed information on the whole proteins. It was revealed that the local structures of proteins at ambient and cryogenic temperatures have obvious differences albeit the overall structures are close to each other. The selected segments under high temperatures are dominated by random coils instead of native helix at ambient and cryogenic temperatures,accompanied by more diversities and faster conversions of conformations. In addition, the temperature or/and the solvent are capable of altering the secondary structures and further the conformations of the segments and the whole proteins. At ambient temperatures, the water solvent helps the proteins to achieve the native conformation with bioactivities. Different from the corresponding segments in proteins, the truncated peptides are endowed with higher structural flexibilities and characterized by random coils even at ambient temperatures, along with the disappearance of helical hydrogen bonds.


Physicochemical characteristics of protein-NP bioconjugates: the role of particle curvature and solution conditions on human serum albumin conformation and fibrillogenesis inhibition.
Gold nanoparticles (Au NPs) from 5 to 100 nm in size synthesized with HAuCl(4) and sodium citrate were complexed with the plasma protein human serum albumin (HSA). Size, surface charge, and surface plasmon bands of the Au NPs are largely modified by the formation of a protein corona via electrostatic interactions and hydrogen bonding as revealed by thermodynamic data. Negative values of the entropy of binding suggested a restriction in the biomolecule mobility upon adsorption. The structure of the adsorbed protein molecules is slightly affected by the interaction with the metal surface, but this effect is enhanced as the NP curvature decreases. Also, it is observed that the protein molecules adsorbed onto the NP surface are more resistant to complete thermal denaturation than free protein ones as deduced from the increases in the melting temperature of the adsorbed protein. Differences in the conformations of the adsorbed protein molecules onto small (<40 nm) and large NPs were observed on the basis of ζ-potential data and FTIR spectroscopy, also suggesting a better resistance of adsorbed protein molecules to thermal denaturing conditions. We think this enhanced protein stability is responsible for a reduced formation of HSA amyloid-like fibrils in the presence of small Au NPs under HSA fibrillation conditions.


Diffusion-weighted MR imaging in closed head injury: high correlation with initial glasgow coma scale score and score on modified Rankin scale at discharge.
To determine whether diffusion-weighted magnetic resonance (MR) imaging findings and conventional MR imaging findings correlate with initial Glasgow Coma Scale score and score on modified Rankin scale at discharge. Twenty-six patients (18 male and eight female patients; mean age, 25.2 years; age range, 4-72 years) with diffuse axonal injury were examined with diffusion-weighted MR imaging and with fluid-attenuated inversion recovery, T2-weighted fast spin-echo, and T2*-weighted gradient-echo sequences. All images were evaluated by two neuroradiologists in consensus. Tissue volume with trauma-related signal-intensity abnormality on images from each sequence, number of lesions for each sequence, number of lesions for all sequences, and number of lesions with reduced apparent diffusion coefficient were correlated with scores on Glasgow Coma Scale and modified Rankin scale. Involvement of brainstem, deep gray matter, and corpus callosum were also correlated with clinical scores. Spearman rank correlation coefficients (r) were calculated. The strongest correlation was between signal-intensity abnormality volume on diffusion-weighted images and modified Rankin score (r = 0.772, P <.001). The strength of this correlation did not improve when only volume of lesions with decreased apparent diffusion coefficient was considered. For lesion number, the strongest correlation was between lesion number on images acquired with all sequences and modified Rankin score (r = 0.662, P <.001). For lesion location, the strongest correlation was between lesion location in the corpus callosum and modified Rankin score (r = 0.513, P =.007). Volume of lesions on diffusion-weighted MR images provides the strongest correlation with a score of subacute on modified Rankin scale at discharge. Total lesion number also correlates well with modified Rankin score. In future, diffusion-weighted images may be useful in determining treatment strategies for acute head injury.


A systematic review and meta-analysis on basic psychological need satisfaction, motivation, and well-being in later life: Contributions of self-determination theory.
As the population ages, it is important to understand the factors that contribute to well-being in the elderly. The purpose of this study is to perform a systematic review and meta-analysis of research on well-being among elderly people conducted particularly within the framework of self-determination theory and, more precisely, to study the relationships among basic psychological need satisfaction, motivation, and well-being. Therefore, a systematic search of the literature was conducted using the databases PsycINFO, PsycARTICLES, Academic Onefile, MEDLINE, Science Direct, and ERIC to find studies published in English and French. China National Knowledge Infrastructure (CNKI) was used to find studies published in Chinese. Ultimately, 23 studies (mainly from the United States, Canada, and Europe; no studies published in China were found) in the field of aging were used for the systematic analysis, 17 of which were used for the meta-analysis. The results reported in these publications are relatively congruent with the idea that basic psychological need satisfaction and motivation (autonomous types) are positively associated with positive indicators of well-being (meaning in life, life satisfaction, positive affect, self-esteem, etc.) and negatively associated with negative indicators of well-being (depression, apathy, etc.).


Does kinematics add meaningful information to clinical assessment in post-stroke upper limb rehabilitation? A case report.
[Purpose] The aims of this case study were to: (a) quantify the impairment and activity restriction of the upper limb in a hemiparetic patient; (b) quantitatively evaluate rehabilitation program effectiveness; and (c) discuss whether more clinically meaningful information can be gained with the use of kinematic analysis in addition to clinical assessment. The rehabilitation program consisted of the combined use of different traditional physiotherapy techniques, occupational therapy sessions, and the so-called task-oriented approach. [Subject and Methods] Subject was a one hemiplegic patient. The patient was assessed at the beginning and after 1 month of daily rehabilitation using the Medical Research Council scale, Nine Hole Peg Test, Motor Evaluation Scale for Upper Extremity in Stroke Patients, and Hand Grip Dynamometer test as well as a kinematic analysis using an optoelectronic system. [Results] After treatment, significant improvements were evident in terms of total movement duration, movement completion velocity, and some smoothness parameters. [Conclusion] Our case report showed that the integration of clinical assessment with kinematic evaluation appears to be useful for quantitatively assessing performance changes.


Subcortical Facilitation of Behavioral Responses to Threat.
Behavioral responses to threat are critical to survival. Several cortical and subcortical brain regions respond selectively to threat. However, the relation of these neural responses and their underlying representations to behavior is unclear. We examined the contribution of lower-order subcortical representations to behavioral responses to threat in adult humans. In Experiments 1 and 2, participants viewed pairs of images presented to the same eye or to different eyes. We observed a monocular advantage, which indicates subcortical facilitation, for ancestral threats (snakes, spiders), but not for modern threats, positive images, or neutral images. In Experiment 3, we presented pairs of snakes or neutral images into the temporal or nasal hemifield. For snakes only, we observed a temporal hemifield advantage, which indicates facilitation by the retino-tectal subcortical pathway. These results advance the current understanding of processing of threat by adult humans by revealing the characteristics of behaviors driven by a lower-order neural mechanism that is specialized for the processing of ancestral threats. The results also contribute to ongoing debates concerning the biological generality of neural mechanisms for processing of complex, emotionally-relevant stimuli by providing evidence for conservation of lower-order neural mechanisms for processing of ancestral threats across both ontogeny and phylogeny.


Human Metapneumovirus Establishes Persistent Infection in Lung Microvascular Endothelial Cells and Primes a Th2-Skewed Immune Response.
Human Metapneumovirus (HMPV) is a major cause of lower respiratory tract infections. HMPV infection has been hypothesized to alter dendritic cell (DC) immune response; however, many questions regarding HMPV pathogenesis within the infected lung remain unanswered. Here, we show that HMPV productively infects human lung microvascular endothelial cells (L-HMVECs). The release of infectious virus occurs for up to more than 30 days of culture without producing overt cytopathic effects and medium derived from persistently HMPV-infected L-HMVECs (secretome) induced monocyte-derived DCs to prime naïve CD4 T-cells toward a Th2 phenotype. Moreover, we demonstrated that infected secretomes trigger DCs to up-regulate OX40L expression and OX40L neutralization abolished the pro-Th2 effect that is induced by HMPV-secretome. We clarified secretome from HMPV by size exclusion and ultracentrifugation with the aim to characterize the role of viral particles in the observed pro-Th2 effect. In both cases, the percentage of IL-4-producing cells and expression of OX40L returned at basal levels. Finally, we showed that HMPV, per se, could reproduce the ability of secretome to prime pro-Th2 DCs. These results suggest that HMPV, persistently released by L-HMVECs, might take part in the development of a skewed, pro-Th2 lung microenvironment.


Maternal, neonatal and community factors influencing neonatal mortality in Brazil.
Child mortality (the mortality of children less than five years old) declined considerably in the developing world in the 1990s, but infant mortality declined less. The reductions in neonatal mortality were not impressive and, as a consequence, there is an increasing percentage of infant deaths in the neonatal period. Any further reduction in child mortality, therefore, requires an understanding of the determinants of neonatal mortality. 209,628 birth and 2581 neonatal death records for the 1998 birth cohort from the city of São Paulo, Brazil, were probabilistically matched. Data were from SINASC and SIM, Information Systems on Live Births and Deaths of Brazil. Logistic regression was used to find the association between neonatal mortality and the following risk factors: birth weight, gestational age, Apgar scores at 1 and 5 minutes, delivery mode, plurality, sex, maternal education, maternal age, number of prior losses, prenatal care, race, parity and community development. Infants of older mothers were less likely to die in the neonatal period. Caesarean delivery was not found to be associated with neonatal mortality. Low birth weight, pre-term birth and low Apgar scores were associated with neonatal death. Having a mother who lives in the highest developed community decreased the odds of neonatal death, suggesting that factors not measured in this study are behind such association. This result may also indicate that other factors over and above biological and more proximate factors could affect neonatal death.


Parental unemployment during the Great Recession and childhood adiposity.
The incidence of adiposity in the early years of life has outgrown the prevalence in older children and adolescents globally; however, the factors influencing weight gain are predominantly studied in adults. This study examines the relationship between changing economic conditions during the Irish recession and child weight in a nationally representative sample of 10,011 Irish children studied at 9 months, 3 years and 5 years old. This study is the first to use longitudinal anthropometric measurements to estimate the impact of direct parental unemployment on children's weight. Fixed effect logistic regression is used to examine the effects of parental unemployment on weight using the Growing up in Ireland infant cohort from 2008 to 2013. Weight and length/height measured by trained interviewers and child growth charts are used to quantify children's weight status according to BMI-for-age and weight-for-age measures. For BMI-for-age, the probability of a child being classified as overweight/obese is 8 percentage points higher if either parent experiences unemployment. A sensitivity analysis of weight-for-age indicates that the probability is of similar magnitude across several alternative growth charts and definitions of adiposity (the WHO standard, British Growth Reference, and Centers for Disease Control). The analysis is repeated, cross-sectionally, for physical activity and diet to clarify mechanisms of effect. The probability of a child consuming healthy food and physical activity with an implied cost is lower if either parent becomes unemployed. A focus on overweight/obesity in the early years is of crucial importance as if current trends are not addressed, a generation of children may grow up with a higher level of chronic disease.


Neuron-specific gene NSG1 binds to and positively regulates sortilin ectodomain shedding via a metalloproteinase-dependent mechanism.
Increasing evidence suggests that aberrant regulation of sortilin ectodomain shedding can contribute to amyloid-β pathology and frontotemporal dementia, although the mechanism by which this occurs has not been elucidated. Here, we probed for novel binding partners of sortilin using multiple and complementary approaches and identified two proteins of the neuron-specific gene (NSG) family, NSG1 and NSG2, that physically interact and colocalize with sortilin. We show both NSG1 and NSG2 induce subcellular redistribution of sortilin to NSG1- and NSG2-enriched compartments. However, using cell surface biotinylation, we found only NSG1 reduced sortilin cell surface expression, which caused significant reductions in uptake of progranulin, a molecular determinant for frontotemporal dementia. In contrast, we demonstrate NSG2 has no effect on sortilin cell surface abundance or progranulin uptake, suggesting specificity for NSG1 in the regulation of sortilin cell surface expression. Using metalloproteinase inhibitors and A disintegrin and metalloproteinase 10 KO cells, we further show that NSG1-dependent reduction of cell surface sortilin occurred via proteolytic processing by A disintegrin and metalloproteinase 10 with a concomitant increase in shedding of sortilin ectodomain to the extracellular space. This represents a novel regulatory mechanism for sortilin ectodomain shedding that is regulated in a neuron-specific manner. Furthermore, this finding has implications for the development of strategies for brain-specific regulation of sortilin and possibly sortilin-driven pathologies.


Pre- and perinatal risk factors for autism spectrum disorder in a New Jersey cohort.
This study evaluated the prevalence of pre- and perinatal risk factors in a cohort of children with autism spectrum disorders compared with the New Jersey population. Our cohort included 268 individuals with an autism spectrum disorder. Birth histories were obtained by a self-administered questionnaire. The autism spectrum disorders cohort rates of 7 perinatal risk factors were significantly higher than New Jersey state rates: mother's age 35 years or older, low birth weight, multiple gestation, prematurity, vaginal bleeding, prolonged labor, and hypoxia. Analysis of clustering of risk factors in the cohort showed no significant differences across maternal and paternal age groups. Older mothers in the cohort had a higher risk of infant hypoxia. Multiple risk factors during pregnancy appear to be associated with a higher risk of autism spectrum disorders in offspring, supporting the hypothesis that environmental influences in conjunction with genetics contribute to the causes of autism spectrum disorders.


Interferon-lambda contributes to innate immunity of mice against influenza A virus but not against hepatotropic viruses.
Virus-infected cells secrete a broad range of interferon (IFN) subtypes which in turn trigger the synthesis of antiviral factors that confer host resistance. IFN-alpha, IFN-beta and other type I IFNs signal through a common universally expressed cell surface receptor, whereas IFN-lambda uses a distinct receptor complex for signaling that is not present on all cell types. Since type I IFN receptor-deficient mice (IFNAR1(0/0)) exhibit greatly increased susceptibility to various viral diseases, it remained unclear to which degree IFN-lambda might contribute to innate immunity. To address this issue we performed influenza A virus infections of mice which carry functional alleles of the influenza virus resistance gene Mx1 and which, therefore, develop a more complete innate immune response to influenza viruses than standard laboratory mice. We demonstrate that intranasal administration of IFN-lambda readily induced the antiviral factor Mx1 in mouse lungs and efficiently protected IFNAR1(0/0) mice from lethal influenza virus infection. By contrast, intraperitoneal application of IFN-lambda failed to induce Mx1 in the liver of IFNAR1(0/0) mice and did not protect against hepatotropic virus infections. Mice lacking functional IFN-lambda receptors were only slightly more susceptible to influenza virus than wild-type mice. However, mice lacking functional receptors for both IFN-alpha/beta and IFN-lambda were hypersensitive and even failed to restrict usually non-pathogenic influenza virus mutants lacking the IFN-antagonistic factor NS1. Interestingly, the double-knockout mice were not more susceptible against hepatotropic viruses than IFNAR1(0/0) mice. From these results we conclude that IFN-lambda contributes to inborn resistance against viral pathogens infecting the lung but not the liver.


Differential Activation of Infiltrating Monocyte-Derived Cells After Mild and Severe Traumatic Brain Injury.
Microglia are the resident innate immune cells of the brain. Although embryologically and functionally distinct, they are morphologically similar to peripheral monocyte-derived cells, resulting in a poor ability to discriminate between the two cell types. The purpose of this study was to develop a rapid and reliable method to simultaneously characterize, quantify, and discriminate between whole populations of myeloid cells from the brain in a murine model of traumatic brain injury. Male C57BL/6 mice underwent traumatic brain injury (n = 16) or sham injury (n = 14). Brains were harvested at 24 h after injury. Multiparameter flow cytometry and sequential gating analysis were performed, allowing for discrimination between microglia and infiltrating leukocytes as well as for the characterization and quantification of individual subtypes within the infiltrating population. The proportion of infiltrating leukocytes within the brain increased with the severity of injury, and the predominant cell types within the infiltrating population were monocyte derived (P = 0.01). In addition, the severity of injury altered the overall makeup of the infiltrating monocyte-derived cells. In conclusion, we describe a flow cytometry-based technique for gross discrimination between infiltrating leukocytes and microglia as well as the ability to simultaneously characterize and quantify individual myeloid subtypes and their maturation states within these populations.


[Nucleotide sequence of gene 31 of the T4 bacteriophage].
We have determined the nucleotide sequence of a region of 656 nucleotides comprising the 31 gene of bacteriophage T4. The coding region consisted of 333 nucleotides directing the synthesis of a polypeptide of 111 amino acids, with a calculated molecular weight of 12,060. The upstream sequence contains the consensus sequences for T4 early and two middle promoters. The downstream sequence contains the consensus sequence for T4 late promoter and the inverted repeats. In addition, there are two incomplete open reading frames in the sequenced region.


[Encounter of cancer cells with bone. The significance of cancer stem cells and epithelial-mesenchymal transition in tumor invasion and metastasis].
Cancer stem cells (CSCs) , which are subset of tumor cells resistant to radiation and chemotherapy, are associated with malignant characteristics of tumor and possess both self-renewal ability and pluripotency for tumor formation. In the process of generating non-CSCs from CSCs, gene mutations and epigenetic changes are induced in those cells, resulting in composition of tumor tissue with heterogeneous cell population. CSCs have been recognized as the source of metastatic foci. Epithelial-mesenchymal transition (EMT) is a change in cellular phenotype characterized by the loss of cell-to-cell adhesions and the gain of migratory behaviors,which has been shown to be a critical factor for initiating cancer invasion and metastasis. However, some recent studies suggest that EMT is not essential requirement for tumor invasion and metastasis. Herein, we discuss the biological significance of CSCs and EMT in tumor invasion and metastasis.


[The estimation of premorbid intelligence levels in French speakers].
Knowledge of cognitive performance earlier in life is essential in order to characterize precisely the extent to which these abilities have declined when an individual is diagnosed as having a dementing illness. The National Adult Reading Test (NART) was developed by Nelson and O'Connell to estimate premorbid intellectual ability in patients suffering from intellectual deterioration due to dementia. The test consists of 50 words, graded in difficulty, whose pronunciation cannot be determined from their spelling. The ability to successfully read irregularly spelt words is relatively robust in the face of current cognitive impairment and is a sensitive marker of intellectual attainment. Because the NART relies on orthographic irregularities in the English language, the construction of analogues of the test in other languages is not simply a matter of translation of the test content. Rather, words in the target language that have comparable properties to those in the NART must be sought. A French adaptation of the NART--the fNART--was developed by Bovet and calibrated on a small French-speaking Swiss sample. In a sample of 30 nondemented subjects, number of words pronounced correctly correlated highly with WAIS-R verbal and total IQ scores and less strongly with performance IQ (r = 0.43). Data available from an epidemiological survey undertaken in Geneva, Switzerland provided an opportunity to establish the measurement properties and construct validity of the fNART in a large sample unselected with respect to cognitive decline. In addition to the fNART, the survey incorporated a brief test battery assessing the domains of crystallized intelligence, memory and cognitive speed. An interview that enabled the diagnosis of dementia according to DSM IV criteria, the Mini Mental State Examination and the Psychogeriatric Assessment Scales (PAS) were also administered. If the fNART measures intellectual ability, substantial correlations between it and the test battery would be expected. Further validation of the test was sought by exploring its relation with years of education. The stability of the fNART was assessed by comparing the scores of subjects with and without dementia, and by examining the relationship of fNART scores to an informant-based report of change in cognitive performance from earlier in life assessed in the PAS. If the fNART is stable in the face of cognitive deterioration, no between-group differences or association with reported cognitive change would be expected. Subjects were randomly selected from residents of the canton of Geneva aged over 65 years. The analyses reported here were undertaken on a sample of 368 persons who gave codable responses to at least 90% of the fNART items. They ranged in age from 65 to 94 years. Subjects were interviewed in their homes by trained lay interviewers. Cronbach's alpha for the forty-item scale was high (0.89). The percentage of subjects correctly pronouncing words ranged from 7.3% for "chamsin" to 96.7% for "agenda". Item response theory (IRT) models were fitted to the data. In a three-parameter model the value of the guessing (asymptote) parameter was vanishing small for all items. Accordingly, a two-parameter model was adopted. The discriminating power (slope) of items ranged considerably from 0.281 (rébus) to 1.192 (béotien). The average slope was 0.656. This corresponds to average factor loading of 0.528 (range 0.270 to 0.766.) The items measure a broad range of ability (mean threshold--0.719, sd = 1.540). Most items, however, discriminate at moderate levels. The parameter values obtained in the current study were compared to those estimated in a French sample of persons at risk of dementia . The correlation between item pairs for slope and parameter estimates was 0.53 and 0.70 respectively. This indicated substantial concordance between the samples regarding the difficulty of the items, but some differences in the power of groups to differentiate ability. In particular, a small number of words that performed very well in the "at risk" sample showed more moderate discrimination in the current study. Scores on the fNART were correlated with measures of crystallised intelligence, memory and cognitive speed. All correlations were statistically significant. With all tests entered a regression equation the multiple correlation coefficient was 0.63. Mean fNART scores of those suffering from DSM IV dementia and those meeting only Criterion A (multiple cognitive deficits) were lower than those of subjects meeting neither set of criteria. However subjects in the first two groups were older than subjects in the undemented group and had significantly lower educational attainment. When these two factors were controlled in an analysis of covariance, the magnitude of the differences between the groups, while still overall significantly different, was substantially reduced. A similar pattern of results applied when psychometric measures of cognitive state--the MMSE and the PAS Cognitive Impairment Scale--were used instead of diagnostic categories. The partial correlations of the fNART with the MMSE and PAS cognitive impairment scale controlling for age and education were 0.25 (P < 0.01) and -0.33 (P < 0.01) respectively. fNART scores did not differ between the sexes, nor were they significantly correlated with PAS Depression, Stroke or Behaviour Change scales. There was a small but significant correlation between the fNART and informant-assessed Cognitive Decline on the PAS. This study demonstrated the excellent measurement properties of a French adaptation of the National Adult Reading Test in a large probability sample of elderly native speakers and provided the first large-sample evidence to support the validity of the fNART as a test of intellectual functioning relatively robust to dementia status. The negligible values of the pseudo-guessing parameters suggest that the goal of choosing words whose pronunciation is not susceptible to guessing has been achieved. The average item discriminability was high and the words used covered the spectrum of ability. The finding of substantial relationships of cognitive performance and educational attainment with fNART scores is important in validating the test as a measure of premorbid cognitive ability. The low correlations of the fNART with informant-based assessment of cognitive decline and age support the fNART as being relative robust to decline in ability. The relationships observed in this French adaptation are comparable to those reported for the English instrument . However, subjects meeting DSM IV criteria for dementia or Criterion A only had lower scores than other subjects. Decline in NART scores with dementia has been observed, particularly in moderate and severe cases. Given that the mechanism of the fNART is the same as the NART it is to be expected that while generally robust to current dementia status, some decline in performance will occur with the progression of the disease. The relationships between the fNART and PAS scales was remarkably similar to those reported by Jorm et al. in an English-speaking sample between the PAS and NART. Although small, the correlation between the fNART and the PAS Cognitive Decline scale might have been expected to be non-significant if the measure were truly stable in the face of intellectual deterioration. However this correlation is mirrored in the original English instruments and may reflect the higher risk of dementia in persons of lower intellectual ability. Further research is desirable to improve the precision of the calibration of the scale against the WAIS-R. Nevertheless, this study has demonstrated that the fNART is a reliable and valid method of assessing premorbid intellectual ability in French speakers.


Vitamin C: an essential "stress hormone" during sepsis.
The stress response is a preserved evolutionary response that functions to enhance the survival of the species. In mammals, the stress response is characterized by activation of the HPA axis and sympathoadrenal system (SAS) as well as the increased synthesis and secretion of vitamin C. Cortisol, catecholamines, and vitamin C act synergistically to increase hemodynamic reserve, maintain immune function and protect the host against excessive oxidant injury. Humans (and anthropoid apes) have lost the ability to synthesize vitamin C and therefore have an impaired stress response. The inability to produce vitamin C has serious implications in septic humans. Treatment with vitamin C appears to restore the stress response and improve the survival of stressed humans.


Data Characterization of Medicaid: Legacy and New Data Formats in the CMS Virtual Research Data Center.
Medicaid is a significant health insurance plan providing healthcare coverage to up to a third of the population of the United Sates. We describe two different formats of Medicaid data within Center for Medicare and Medicaid Services Virtual Research Data Center. We analyze record length, age and enrollment justification among patients for both data formats. As of December 2016, the total size of Medicaid population available from CMS is 92,953,389; 45% of patients are aged 0 to 18, 26.6% are aged 19-35 and 23.2% are aged 36-64. In terms of Medicaid eligibility, 35.6% qualify due to (child) age and 26.8% qualify due to income. We also compare the volume of Medicaid to Medicare for year 2016. We conclude that Medicaid data includes patients with significant record lengths and relatively well documented enrollment justification, which are high value assets for data reuse researchers that are willing to balance known data limitations with careful analysis design and interpretation.


Immunomodulatory effect of Eriobotrya japonica seed extract on allergic dermatitis rats.
We examined the immunomodulatory effect of Eriobotrya japonica seed extract (ESE) on rat allergic dermatitis elicited by repeated dinitrofluorobenzene (DNFB) application on the ear. Oral administration of ESE significantly inhibited development of allergic dermatitis based on lower ear thickness and serum immunoglobulin E (IgE) levels. Th1 cytokine interferon-gamma (IFN-gamma) and interleukin-2 (IL-2), Th2 cytokine interleukin-4 (IL-4) and interleukin-10 (IL-10) in the lesional skin were determined. Oral administration of ESE significantly decreased IL-4 while significantly increasing IL-10 in lesional skin, and the lower levels of IFN-gamma and IL-2 were reversed by oral administration of ESE. The infiltration of eosinophils in the lesional skin was decreased by oral administration of ESE. These results suggested that ESE exerts anti-allergic actions by improving the balance of Th1/Th2 in allergic dermatitis.


Immunological aspects of chronic fatigue syndrome.
Chronic fatigue syndrome (CFS) is a specific clinical condition that characterises unexplained disabling fatigue and a combination of non-specific accompanying symptoms for at least 6 months, in the absence of a medical diagnosis that would otherwise explain the clinical presentation. Other common symptoms include headaches, myalgia, arthralgia, and post-exertional malaise; cognitive difficulties, with impaired memory and concentration; unrefreshing sleep; and mood changes. Similar disorders have been described for at least two centuries and have been differently named neurasthenia, post-viral fatigue, myalgic encephalomyelitis and chronic mononucleosis. Recent longitudinal studies suggest that some people affected by chronic fatigue syndrome improve with time but that most remain functionally impaired for several years. The estimated worldwide prevalence of CFS is 0.4-1% and it affects over 800,000 people in the United States and approximately 240,000 patients in the UK. No physical examination signs are specific to CFS and no diagnostic tests identify this syndrome. The pathophysiological mechanism of CFS is unclear. The main hypotheses include altered central nervous system functioning resulting from an abnormal immune response against a common antigen; a neuroendocrine disturbance; cognitive impairment caused by response to infection or other stimuli in sentient people. The current concept is that CFS pathogenesis is a multifactorial condition. Various studies have sought evidence for a disturbance in immunity in people with CFS. An alteration in cytokine profile, a decreased function of natural killer (NK) cells, a presence of autoantibodies and a reduced responses of T cells to mitogens and other specific antigens have been reported. The observed high level of pro-inflammatory cytokines may explain some of the manifestations such as fatigue and flu-like symptoms and influence NK activity. Abnormal activation of the T lymphocyte subsets and a decrease in antibody-dependent cell-mediated cytotoxicity have been described. An increased number of CD8+ cytotoxic T lymphocytes and CD38 and HLA-DR activation markers have been reported, and a decrease in CD11b expression associated with an increased expression of CD28+ T subsets has been observed. This review discusses the immunological aspects of CFS and offers an immunological hypothesis for the disease processes.


Prognostic significance of NPM1 mutations in the absence of FLT3-internal tandem duplication in older patients with acute myeloid leukemia: a SWOG and UK National Cancer Research Institute/Medical Research Council report.
Younger patients with acute myeloid leukemia (AML) harboring NPM1 mutations without FLT3-internal tandem duplications (ITDs; NPM1-positive/FLT3-ITD-negative genotype) are classified as better risk; however, it remains uncertain whether this favorable classification can be applied to older patients with AML with this genotype. Therefore, we examined the impact of age on the prognostic significance of NPM1-positive/FLT3-ITD-negative status in older patients with AML. Patients with AML age ≥ 55 years treated with intensive chemotherapy as part of Southwest Oncology Group (SWOG) and UK National Cancer Research Institute/Medical Research Council (NCRI/MRC) trials were evaluated. A comprehensive analysis first examined 156 patients treated in SWOG trials. Validation analyses then examined 1,258 patients treated in MRC/NCRI trials. Univariable and multivariable analyses were used to determine the impact of age on the prognostic significance of NPM1 mutations, FLT3-ITDs, and the NPM1-positive/FLT3-ITD-negative genotype. Patients with AML age 55 to 65 years with NPM1-positive/FLT3-ITD-negative genotype treated in SWOG trials had a significantly improved 2-year overall survival (OS) as compared with those without this genotype (70% v 32%; P < .001). Moreover, patients age 55 to 65 years with NPM1-positive/FLT3-ITD-negative genotype had a significantly improved 2-year OS as compared with those age > 65 years with this genotype (70% v 27%; P < .001); any potential survival benefit of this genotype in patients age > 65 years was marginal (27% v 16%; P = .33). In multivariable analysis, NPM1-positive/FLT3-ITD-negative genotype remained independently associated with an improved OS in patients age 55 to 65 years (P = .002) but not in those age > 65 years (P = .82). These results were confirmed in validation analyses examining the NCRI/MRC patients. NPM1-positive/FLT3-ITD-negative genotype remains a relatively favorable prognostic factor for patients with AML age 55 to 65 years but not in those age > 65 years.


The very low number of calcium-induced permeability transition pores in the single mitochondrion.
Mitochondrial permeability transition (PT) is a phenomenon of stress-induced increase in nonspecific permeability of the mitochondrial inner membrane that leads to disruption of oxidative phosphorylation and cell death. Quantitative measurement of the membrane permeability increase during PT is critically important for understanding the PT's impact on mitochondrial function. The elementary unit of PT is a PT pore (PTP), a single channel presumably formed by either ATP synthase or adenine nucleotide translocator (ANT). It is not known how many channels are open in a single mitochondrion during PT, which makes it difficult to quantitatively estimate the overall degree of membrane permeability. Here, we used wide-field microscopy to record mitochondrial swelling and quantitatively measure rates of single-mitochondrion volume increase during PT-induced high-amplitude swelling. PT was quantified by calculating the rates of water flux responsible for measured volume changes. The total water flux through the mitochondrial membrane of a single mitochondrion during PT was in the range of (2.5 ± 0.4) × 10-17 kg/s for swelling in 2 mM Ca2+ and (1.1 ± 0.2) × 10-17 kg/s for swelling in 200 µM Ca2+. Under these experimental conditions, a single PTP channel with ionic conductance of 1.5 nS could allow passage of water at the rate of 0.65 × 10-17 kg/s. Thus, we estimate the integral ionic conductance of the whole mitochondrion during PT to be 5.9 ± 0.9 nS for 2 mM concentration of Ca2+ and 2.6 ± 0.4 nS for 200 µM of Ca2+. The number of PTPs per mitochondrion ranged from one to nine. Due to the uncertainties in PTP structure and model parameters, PTP count results may be slightly underestimated. However, taking into account that each mitochondrion has ∼15,000 copies of ATP synthases and ANTs, our data imply that PTP activation is a rare event that occurs only in a small subpopulation of these proteins.


Sequential loss of suppressor genes for three specific functions during in vivo carcinogenesis.
As normal cells undergo neoplastic transformation, multiple suppressor gene functions are lost or inactivated. However, the relative contribution that individual suppressor gene defects play in the sequential evolution of solid tumors has not yet been evaluated in a readily analyzable experimental model of carcinogenesis. The present study was undertaken to: (a) document the loss of suppressor genes implicated in the control of angiogenic activity, anchorage and serum growth requirements, and proliferative life span, in populations of hamster buccal pouch (HBP) keratinocytes (Kr) initiated in vivo with the chemical carcinogen 7,12 dimethylbenz(a)anthracene and (b), to determine what combination of defective suppressor genes are necessary for tumorigenesis. Kr were isolated from HBPs at various times after treating the mucosal surfaces in vivo with 7,12 dimethylbenz(a)anthracene. Cells or their conditioned culture media were evaluated for: (a) angiogenic activity in vivo and in vitro, (b) anchorage independent growth, (c) growth in low serum, (d) immortality, and (e) tumorigenicity. Angiogenic activity and immortality were the first two phenotypes detected with anchorage independence and tumorigenesis emerging late in the carcinogenic process. Hybrids generated between Kr which were angiogenic, but otherwise negative for all other phenotypic traits, and a transformed and tumorigenic HBP carcinoma cell line, E1-1, were suppressed for all phenotypes except angiogenic activity and none of the hybrids were tumorigenic. In contrast, Kr positive for angiogenic activity, anchorage independence, immortality, and tumorigenicity and hybrids generated between these cells and E1-1 carcinoma, were tumorigenic. However, hybrids between a nontumorigenic, anchorage independent, immortal but nonangiogenic Kr, and the E1-1 line were not tumorigenic. These results suggest that (a) angiogenic activity is an early phenotypic trait that emerges as a result of the loss of a suppressor gene function and (b) loss of this function is essential but not sufficient for the development of HBP tumors.


Local care and treatment of liver disease (LOCATE) - A cluster-randomized feasibility study to discover, assess and manage early liver disease in primary care.
Chronic liver disease is an escalating problem both in the United Kingdom and worldwide. In the UK mortality rates have risen sharply over the previous 50 years predominantly due to alcohol, however the increasing prevalence of non-alcohol related fatty liver disease both in the UK and elsewhere is also of concern. Liver disease develops silently hence early detection of fibrosis is essential to prevent progression. Primary care presents an opportunity to identify at risk populations, however assessment largely comprises of indirect markers of fibrosis which have little prognostic value. We hypothesised that setting up nurse-led primary care based liver clinics using additional non-invasive testing would increase the number of new diagnoses of liver disease compared to usual care. This was a prospective, cluster randomised feasibility trial based in urban primary care in Southampton, United Kingdom. 10 GP practices were randomised to either intervention (liver health nurse) or control (care as usual). Pre recruitment audits were carried out in each practice to ascertain baseline prevalence of liver disease. Participants were subsequently recruited in intervention practices from July 2014-March 2016 via one of 3 pathways: GP referral, nurse led case finding based on risk factors or random AUDIT questionnaire mailouts. Liver assessment included the Southampton Traffic Light test (serum fibrosis markers HA and P3NP) and transient elastography (FibroScan). Cases were ascribed as 'no fibrosis', 'liver warning', 'progressive fibrosis' or 'probable cirrhosis'. Post recruitment audits were repeated and incident liver diagnoses captured from July 2014-September 2016. Each new diagnosis was reviewed in a virtual clinic by a consultant hepatologist. 910 participants were seen in the nurse led clinic-44 (4.8%) probable cirrhosis, 141 (15.5%) progressive fibrosis, 220 (24.2%) liver warning and 505 (55.5%) no evidence of liver fibrosis. 450 (49.5%) cases were due to NAFLD with 356 (39.1%) from alcohol. In the 405 with a liver disease diagnosis, 136 (33.6%) were referred by GP, 218 (53.8%) from nurse led case finding and 51 (12.6%) from the AUDIT mailout. 544 incident cases were identified in the intervention arm compared to 221 in the control arm in the period July 2014-September 2016 (adjusted odds ratio 2.4, 95% CI 2.1 to 2.8). The incorporation of a liver health nurse into GP practices was simple to arrange and yielded a much higher number of new diagnoses of liver disease compared to usual care. Nearly half of all participants recruited had a degree of liver disease. Nurse led case finding and GP referrals were most effective compared to AUDIT questionnaire mailouts in an urban population in identifying unknown disease. Utilising study and previous data allowed quick and effective virtual review by a hepatologist. Identifying those who are at risk of liver disease from harmful alcohol use remains a challenge and needs to be addressed in future work.


Cyclic oscillation of blood neutrophils in a patient with multiple myeloma.
A patient with multiple myeloma developed periodic blood neutropenia (periodicity of 15-25 days) after 3 yr of intermittent treatment with cytotoxic agents. Peaks of serum colony-stimulating activity (CSA) level coincided with valleys of blood neutrophils. Fraction of marrow neutrophils in the multiplicative pool was high during blood neutrophil valleys and low during neutrophil peaks. In contrast, the maturation storage pool exhibited the reverse pattern. An increased fraction of marrow neutrophilic cells in the multiplicative pool was in active proliferation during a blood neutrophil valley and a decreased fraction during a blood neutrophil peak. These findings suggest that the marrow granulopoiesis was regulated through CSA. The defect causing the periodicity was probably related to the reduced number of neutrophils in the marrow maturation storage pool, which in turn may be related to a reduced and/or defective granulocytic stem cell pool size consequent to the long-term administration of cytotoxic drugs and/or infiltration of the marrow by myeloma cells.


Isolation and high-performance thin layer chromatographic estimation of Lupeol from Oxystelma esculentum.
Oxystelma esculentum R. Br. (Family: Asclepiadaceae) is a perennial twiner growing in water-logged areas of the Indian sub-continent. It is used traditionally as a diuretic, laxative and an anti-ulcer agent. However, there is no method available for isolation and estimation of a marker compound from this plant. The present work deals with the isolation and structure elucidation of Lupeol from O. esculentum and high-performance thin layer chromatographic (HPTLC) method development for its estimation. The petroleum ether extract of the entire plant of O. esculentum was subjected to further fractionation followed by pilot thin layer chromatographic (TLC) experiments and column chromatography. This yielded a pure, white, crystalline solid which resolved at Rf 0.65 upon TLC of chloroform fraction of the petroleum ether extract using the mobile phase toluene: methanol (9:1). This compound was subjected to ultraviolet (UV), infrared (IR), gas chromatography-mass spectroscopy (GC-MS) and (1) H-nuclear magnetic resonance ((1)H-NMR) spectral analysis and its structure elucidation revealed it to be Lupeol. A novel HPTLC method for the estimation of Lupeol from O. esculentum was developed, in which it was found to be 0.829 ± 0.09% w/w. The method developed was found to be easy, simple, precise, efficient, accurate, reproducible, specific and sensitive, and could serve as a suitable tool for routine analysis and phytochemical authentication of O. esculentum.


The c-Abl tyrosine kinase regulates actin remodeling at the immune synapse.
Actin dynamics during T-cell activation are controlled by the coordinate action of multiple actin regulatory proteins, functioning downstream of a complex network of kinases and other signaling molecules. The c-Abl nonreceptor tyrosine kinase regulates actin responses in nonhematopoietic cells, but its function in T cells is poorly understood. Using kinase inhibitors, RNAi, and conditional knockout mice, we investigated the role of c-Abl in controlling the T-cell actin response. We find that c-Abl is required for normal actin polymerization and lamellipodial spreading at the immune synapse, and for downstream events leading to efficient interleukin-2 production. c-Abl also plays a key role in signaling chemokine-induced T-cell migration. c-Abl is required for the appropriate function of 2 proteins known to be important for controlling actin responses to T-cell receptor (TCR) engagement, the actin-stabilizing adapter protein HS1, and the Rac1-dependent actin polymerizing protein WAVE2. c-Abl binds to phospho-HS1 via its SH2 domains and is required for full tyrosine phosphorylation of HS1 during T-cell activation. In addition, c-Abl is required for normal localization of WAVE2 to the immune synapse (IS). These studies identify c-Abl as a key player in the signaling cascade, leading to actin reorganization during T-cell activation.


Conditional survival after surgical resection of gastric cancer: a multi-institutional analysis of the us gastric cancer collaborative.
Survival estimates following surgical resection of gastric adenocarcinoma are traditionally reported as survival from the date of surgery. Conditional survival (CS) estimates, however, may be more clinically relevant by accounting for time already survived. We assessed CS following surgical resection for gastric adenocarcinoma. We analyzed 807 patients who underwent resection for gastric adenocarcinoma from 2000 to 2012 at seven participating institutions in the U.S. Gastric Cancer Collaborative. Cox proportional hazards models were used to evaluate factors associated with overall survival. Three-year CS estimates at "x" year after surgery were calculated as follows: CS3 = S(x+3)/S(x). Overall 1-, 3-, and 5-year overall survival rates after gastric resection were 42, 34, and 30 %, respectively. Using CS estimates, the probability of surviving an additional 3 years given that the patient had survived at 1, 3, and 5 years were 56, 71, and 82 %, respectively. Patients with higher risk at baseline (i.e., stage III or IV disease, lymphovascular invasion) demonstrated the greatest increase in CS over time. Survival estimates following surgical resection of gastric adenocarcinoma is dynamic; the probability of survival increases with time already survived. Patients with worse prognostic features at the time of surgery had the greatest increases in CS over time. Conditional survival estimates provide important information about the changing probability of survival over time and should be used among patients with resected gastric adenocarcinoma to guide subsequent follow-up strategies.


Specific pathogen-free status alters immunophenotype in rhesus macaques: implications for the study of simian immunodeficiency virus.
The repertoire of viruses to which research primates are exposed, even in the absence of clinical disease, may contribute to experimental confounding. In this study we examined whether standard specific pathogen-free (SPF) rhesus macaques exposed to a wider spectrum of enzootic viruses and expanded SPF macaques derived to exclude a greater number of viral agents would display alterations in immune activation or immune cell populations. Given the impact of immunophenotype on human immunodeficiency virus (HIV) progression and the importance of the simian immunodeficiency virus (SIV) model for the study of HIV pathogenesis, we elected to additionally examine the impact of SPF status on the capacity of peripheral blood mononuclear cells (PBMCs) to support SIV replication. The expanded SPF group displayed significant immune alterations including increased serum interleukin (IL)-15 and a greater in vitro elaboration of GM-CSF, IL1ra, VEGF, IL-10, IL12/23, and MIP-1b. Consistent with reduced viral antigenic exposure in expanded SPF macaques, decreased CD4(+) and CD8(+) transitional and effector memory (T(EM)) cell populations were observed. Expanded SPF PBMC cultures also demonstrated an increased peak (192.61 ng/ml p27) and area under the curve in in vitro SIV production (1968.64 ng/ml p27) when compared to standard SPF macaques (99.32 ng/ml p27; p=0.03 and 915.17 ng/ml p27; p=0.03, respectively). In vitro SIV replication did not correlate with CD4(+) T(EM) cell counts but was highly correlated with serum IL-15 in the subset of animals examined. Findings suggest that an altered immunophenotype associated with the maintenance of primates under differing levels of bioexclusion has the potential to impact the outcome of SIV studies and models for which the measurement of immunologic endpoints is critical.


"In vitro" capacitation and further progesterone-induced acrosome exocytosis are linked to specific changes in the expression and location of threonine phosphorylation of boar spermatozoa.
The aim of this study was to determine if the achievement of the "in vitro" capacitation (IVC) status and subsequent progesterone-induced "in vitro" acrosome exocytosis (IVAE) was accompanied with overall changes in threonine phosphorylation (pThre) of boar spermatozoa. For this purpose, mono- and bi-dimensional Western blot analyses as well as immunocytochemistry studies against pThre were performed in boar sperm subjected to IVC and subsequent IVAE. Mono-dimensional Western blot in non-capacitated samples showed that launching of IVC did induce an overall increase in signal intensity in all observed bands that was followed by a subsequent decrease afterwards. Bi-dimensional Western blot analysis showed the presence of four main signal protein clusters. The attainment of IVC induced an overall decrease in the number and intensity of spots of Clusters A, B and C and a concomitant increase in the intensity of spots of Cluster D. The IVAE launching caused a rapid increase in the intensity of spots of Clusters B, C and D, which was followed by a subsequent decrease of the intensity together with a concomitant pI displacement of Cluster C. Finally, immunocytochemistry showed that the pThre signal of non-capacitated cells was located at the whole sperm. The IVC did not induce prominent changes in this location. In contrast, the induction of IVAE caused the appearance of an additional an intense acrosome and tail pThre signal that subsequently decreased. In conclusion, our results indicate that IVC and further IVAE induced specific changes in the intensity and appearance of pThre protein phosphorylation which were linked to changes of specific protein characteristics as pI. These results support, thus, the existence of a specific role of pThre in IVC/IVAE of boar sperm.


Genome graphs and the evolution of genome inference.
The human reference genome is part of the foundation of modern human biology and a monumental scientific achievement. However, because it excludes a great deal of common human variation, it introduces a pervasive reference bias into the field of human genomics. To reduce this bias, it makes sense to draw on representative collections of human genomes, brought together into reference cohorts. There are a number of techniques to represent and organize data gleaned from these cohorts, many using ideas implicitly or explicitly borrowed from graph-based models. Here, we survey various projects underway to build and apply these graph-based structures-which we collectively refer to as genome graphs-and discuss the improvements in read mapping, variant calling, and haplotype determination that genome graphs are expected to produce.


Four years of lamivudine treatment in Chinese patients with chronic hepatitis B.
This study assessed the efficacy and safety of up to 4 years of lamivudine treatment and the clinical relevance of the emergence of YMDD-variant hepatitis B virus (HBV). Fifty-eight Chinese adult patients with chronic hepatitis B (CHB) were randomized to lamivudine 100 mg/day for up to 5 years and were monitored for YMDD-variant HBV, hepatitis B e antigen (HBeAg) seroconversion (loss of HBeAg and detectable antibody to HBeAg) and serum alanine aminotransferase (ALT) concentrations. Four-year data are reported here. The rate of HBeAg seroconversion increased with extended therapy and also with higher baseline ALT concentrations. YMDD-variant HBV was detected in 67% (39/58) of patients at some point during treatment. After 4 years, a total of 47% (27/58) of patients achieved HBeAg seroconversion. Thirty-three per cent (13/39) of patients with YMDD-variant HBV achieved HBeAg seroconversion; this increased to 57% (8/14) in patients with moderately elevated (>2-5 x upper limit of normal) pre-treatment ALT concentrations. The proportion of patients that achieved normal serum ALT increased from 29% (17/58) at baseline to 69% (31/45) following 4 years of treatment. That included 68% (23/34) of patients with YMDD-variant HBV and 73% (8/11) of those without the variant. All patients receiving lamivudine had reduced serum concentrations of HBV-DNA compared with baseline, despite the emergence of YMDD-variant HBV in 39 patients. Lamivudine was generally well tolerated; there was little change in the number or type of drug-related adverse events in the fourth year of the study. Despite the emergence of YMDD-variant HBV, Chinese patients showed increased HBeAg seroconversion and improvement in ALT levels with an increased duration of treatment with lamivudine.


The Nicaraguan pediatric dengue cohort study: incidence of inapparent and symptomatic dengue virus infections, 2004-2010.
Dengue, caused by the four serotypes of dengue virus (DENV), is the most prevalent mosquito-borne viral disease of humans. To examine the incidence and transmission of dengue, the authors performed a prospective community-based cohort study in 5,545 children aged 2-14 years in Managua, Nicaragua, between 2004 and 2010. Children were provided with medical care through study physicians who systematically recorded medical consult data, and yearly blood samples were collected to evaluate DENV infection incidence. The incidence of dengue cases observed was 16.1 cases (range 3.4-43.5) per 1,000 person-years (95% CI: 14.5, 17.8), and a pattern of high dengue case incidence every other year was observed. The incidence of DENV infections was 90.2 infections (range 45.2-105.3) per 1,000 person-years (95% CI: 86.1, 94.5). The majority of DENV infections in young children (<6 years old) were primary (60%) and the majority of infections in older children (≥ 9 years of age) were secondary (82%), as expected. The incidence rate of second DENV infections (121.3 per 1,000 person-years; 95% CI: 102.7, 143.4) was significantly higher than the incidence rate of primary DENV infections (78.8 per 1,000 person-years; 95% CI: 73.2, 84.9). The rigorous analytic methodology used in this study, including incidence reporting in person-years, allows comparison across studies and across different infectious diseases. This study provides important information for understanding dengue epidemiology and informing dengue vaccine policy.


Body composition indices as predictors of humoral response to mRNA vaccination against COVID-19 in patients on maintenance hemodialysis.
Patients on maintenance hemodialysis show lower serological response to mRNA vaccines. Main causes that contribute to this phenomenon are uremic milieu and older age. However, there are no data on the impact of body composition parameters to humoral response. In this retrospective study, we used data from adult patients on maintenance hemodialysis who received vaccination with 2 doses of BNT162b2. Quantitative determination of antibodies to SARS-CoV-2 spike (S) protein receptor binding domain was performed using the Elecsys immunoassay. Antibody levels higher than 0.8 and 264 U/mL were considered positive and protective, respectively. Body composition parameters were assessed using multifrequency bioelectrical impedance spectroscopy. Overall, 49 patients were included in the study. Three weeks after the 1st vaccination, 34% of patients, and 3 weeks and 3 months after the 2nd vaccination, 100% of patients had detectable titers. Protective titer was developed in 43% of patients 3 weeks after the 2nd vaccination and then decreased to 24% 3 months after the 2nd vaccination. More years on dialysis were correlated to the absence of protective titers. Higher prediction marker values correlated to poor antibody response, and phase angle was negatively associated with the development of protective titers. Patients with protective titers at 3 months after the 2nd vaccination had significantly lower prediction marker and higher phase angle values. Parameters of body composition correlate and affect antibody response in patients on hemodialysis. The main observation is that immunogenicity of mRNA vaccines is influenced by phase angle and prediction marker.


Tracking SARS-CoV-2 Omicron diverse spike gene mutations identifies multiple inter-variant recombination events.
The current pandemic of COVID-19 is fueled by more infectious emergent Omicron variants. Ongoing concerns of emergent variants include possible recombinants, as genome recombination is an important evolutionary mechanism for the emergence and re-emergence of human viral pathogens. In this study, we identified diverse recombination events between two Omicron major subvariants (BA.1 and BA.2) and other variants of concern (VOCs) and variants of interest (VOIs), suggesting that co-infection and subsequent genome recombination play important roles in the ongoing evolution of SARS-CoV-2. Through scanning high-quality completed Omicron spike gene sequences, 18 core mutations of BA.1 (frequency >99%) and 27 core mutations of BA.2 (nine more than BA.1) were identified, of which 15 are specific to Omicron. BA.1 subvariants share nine common amino acid mutations (three more than BA.2) in the spike protein with most VOCs, suggesting a possible recombination origin of Omicron from these VOCs. There are three more Alpha-related mutations in BA.1 than BA.2, and BA.1 is phylogenetically closer to Alpha than other variants. Revertant mutations are found in some dominant mutations (frequency >95%) in the BA.1. Most notably, multiple characteristic amino acid mutations in the Delta spike protein have been also identified in the "Deltacron"-like Omicron Variants isolated since November 11, 2021 in South Africa, which implies the recombination events occurring between the Omicron and Delta variants. Monitoring the evolving SARS-CoV-2 genomes especially for recombination is critically important for recognition of abrupt changes to viral attributes including its epitopes which may call for vaccine modifications.


Plant programmed cell death caused by an autoactive form of Prf is suppressed by co-expression of the Prf LRR domain.
In tomato, the NBARC-LRR resistance (R) protein Prf acts in concert with the Pto or Fen kinase to determine immunity against Pseudomonas syringae pv. tomato (Pst). Prf-mediated defense signaling is initiated by the recognition of two sequence-unrelated Pst-secreted effector proteins, AvrPto and AvrPtoB, by tomato Pto or Fen. Prf detects these interactions and activates signaling leading to host defense responses including localized programmed cell death (PCD) that is associated with the arrest of Pst growth. We found that Prf variants with single amino acid substitutions at D1416 in the IHD motif (isoleucine-histidine-aspartic acid) in the NBARC domain cause effector-independent PCD when transiently expressed in leaves of Nicotiana benthamiana, suggesting D1416 plays an important role in activation of Prf. The N-terminal region of Prf (NPrf) and the LRR domain are required for this autoactive Prf cell death signaling but dispensable for accumulation of the Prf(D1416V) protein. Significantly, co-expression of the Prf LRR but not NPrf, with Prf(D1416V), AvrPto/Pto, AvrPtoB/Pto, an autoactive form of Pto (Pto(Y207D)), or Fen completely suppresses PCD. However, the Prf LRR does not interfere with PCD caused by Rpi-blb1(D475V), a distinct R protein-mediated PCD signaling event, or that caused by overexpression of MAPKKKα, a protein acting downstream of Prf. Furthermore, we found the Prf(D1416V) protein is unable to accumulate in plant cells when co-expressed with the Prf LRR domain, likely explaining the cell death suppression. The mechanism for the LRR-induced degradation of Prf(D1416V) is unknown but may involve interference in the intramolecular interactions of Prf or to binding of the unattached LRR to other host proteins that are needed for Prf stability.


Effect of patient arm motion in whole-body PET/CT.
Arm motion during whole-body PET/CT acquisition is not uncommon and can give rise to striking cold artifacts on PET images. We investigated the mechanisms that underlie these artifacts and proposed a potential solution. A phantom experiment based on 5 clinical cases of suspected arm motion was designed. The experiment involved a central 20-cm-diameter (68)Ge/(68)Ga cylinder simulating the neck and 2 peripheral 10-cm-diameter (18)F cylinders simulating arms. After motion-free CT and PET on a whole-body PET/CT system, the position of the arms was altered so as to introduce different amounts of misalignment. Twenty sequential PET scans were acquired in this position, alternating between 2-dimensional (2D) and 3-dimensional (3D) acquisition, as the (18)F decayed. Decay of (18)F in the arms, while the activity in the (68)Ge/(68)Ga cylinder remained approximately constant, allowed the relative impact of scatter and attenuation-correction errors to be determined. Image artifacts were largely confined to the local region of motion in 2D but extended throughout the affected slices in 3D, where they manifested as a striking underestimation of radiotracer concentration that became more significant with increasing misalignment. For 3D, scatter-correction error depended on activity in the arms, but for typical activity concentrations scatter-correction error was more significant than attenuation-correction error. 3D image reconstruction without scatter correction substantially eliminated these artifacts in both phantom and patient images. Reconstruction artifacts due to patient arm motion can be substantial and should be recognized because they can affect both qualitative and quantitative assessment of PET.


Comparison of two approaches for measuring household wealth via an asset-based index in rural and peri-urban settings of Hunan province, China.
There are growing concerns regarding inequities in health, with poverty being an important determinant of health as well as a product of health status. Within the People's Republic of China (P.R. China), disparities in socio-economic position are apparent, with the rural-urban gap of particular concern. Our aim was to compare direct and proxy methods of estimating household wealth in a rural and a peri-urban setting of Hunan province, P.R. China. We collected data on ownership of household durable assets, housing characteristics, and utility and sanitation variables in two village-wide surveys in Hunan province. We employed principal components analysis (PCA) and principal axis factoring (PAF) to generate household asset-based proxy wealth indices. Households were grouped into quartiles, from 'most wealthy' to 'most poor'. We compared the estimated household wealth for each approach. Asset-based proxy wealth indices were compared to those based on self-reported average annual income and savings at the household level. Spearman's rank correlation analysis revealed that PCA and PAF yielded similar results, indicating that either approach may be used for estimating household wealth. In both settings investigated, the two indices were significantly associated with self-reported average annual income and combined income and savings, but not with savings alone. However, low correlation coefficients between the proxy and direct measures of wealth indicated that they are not complementary. We found wide disparities in ownership of household durable assets, and utility and sanitation variables, within and between settings. PCA and PAF yielded almost identical results and generated robust proxy wealth indices and categories. Pooled data from the rural and peri-urban settings highlighted structural differences in wealth, most likely a result of localized urbanization and modernization. Further research is needed to improve measurements of wealth in low-income and transitional country contexts.


Effect of sodium arsenite dose administered in the drinking water on the urinary bladder epithelium of female arsenic (+3 oxidation state) methyltransferase knockout mice.
The enzyme arsenic (+3 oxidation state) methyltransferase (As3mt) catalyzes reactions converting inorganic arsenic to methylated metabolites, some of which are highly cytotoxic. In a previous study, female As3mt knockout (KO) mice treated with diet containing 100 or 150 ppm arsenic as arsenite showed systemic toxicity and significant effects on the urothelium. In the present study, we showed that the cytotoxic and proliferative effects of arsenite administration on the urothelium are dose dependent. Female wild-type C57BL/6 mice and As3mt KO mice were divided into five groups (n = 7) with free access to drinking water containing 0, 1, 10, 25, or 50 ppm arsenic as arsenite for 4 weeks. At sacrifice, urinary bladders of both As3mt KO and wild-type mice showed hyperplasia by light microscopy; however, the hyperplasia was more severe in the As3mt KO mice. Intracytoplasmic granules were detected in the urothelium of As3mt KO and wild-type mice at arsenic doses ≥ 10 ppm but were more numerous, more extensive, and larger in the KO mice. A no effect level for urothelial effects was identified at 1 ppm arsenic in the wild-type and As3mt KO mice. In As3mt KO mice, livers showed mild acute inflammation and kidneys showed hydronephrosis. The present study shows a dose-response for the effects of orally administered arsenite on the bladder urothelium of wild-type and As3mt KO mice, with greater effects in the KO strain but with a no effect level of 1 ppm for both.


Coexistence of spin density waves and superconductivity in (TMTSF)2PF6.
We present simultaneous measurements of angular-dependent magnetoresistance and thermopower along all three crystal axes in (TMTSF)2PF6 for pressures to 7.4 kbar and magnetic fields to 35 T. (TMTSF)2PF6 under pressure shows the coexistence of spin density wave and metal-superconducting orders. We suggest that this coexistence results neither in microscopic coexistence nor in a new soliton wall phase, contrary to previous suggestions, but in phase separation into domains of the high-pressure metal and the low-pressure spin density wave phases. Simultaneous measurement of transport along all crystal axes allows us to unambiguously describe the domain structure, whereas the superconducting transition temperature and four independent Fermi surface-sensitive magnetoresistance signatures allow us to unambiguously characterize the coexisting metallic domains.


Transient elevation of corticosterone alters begging behavior and growth of white-crowned sparrow nestlings.
Developing animals may face a cost-benefit tradeoff during growth mediated through hormones such as glucocorticoids, as the hormone is essential for development but can have detrimental consequences. To investigate potential tradeoffs caused by brief, moderate elevations of corticosterone in avian young, we artificially elevated the hormone levels in two ways: feeding corticosterone-containing worms and applying corticosterone dermal patches. The former experiment tested the effects of an acute corticosterone elevation (25 min) on begging behavior, whereas the latter explored the effects of artificially elevated corticosterone for 24 to 48 h on growth. Corticosterone altered both begging behavior and growth of white-crowned sparrow nestlings. It increased latency to beg immediately after the treatment and suppressed growth as early as 24 h after the patch application. These experiments also showed that the effects depended on the age or types of development (e.g. gaining mass or growing feathers) that the nestlings were going through.


Neurocognitive and Quality of Life Improvements Associated With Aerobic Training for Individuals With Persistent Symptoms After Mild Traumatic Brain Injury: Secondary Outcome Analysis of a Pilot Randomized Clinical Trial.
Objective: To report secondary neurocognitive and quality of life outcomes for a pilot randomized clinical trial (RCT) of aerobic training for management of prolonged symptoms after a mild traumatic brain injury (mTBI) in adolescents. Setting: Outpatient research setting. Participants: Thirty adolescents between the ages of 12 and 17 years who sustained a mTBI and had between 4 and 16 weeks of persistent post-concussive symptoms. Design: Secondary outcome analysis of a partially masked RCT of sub-symptom exacerbation aerobic training compared with a full-body stretching program highlighting cognitive and quality of life outcomes. Main Measures: The secondary outcomes assessed included neurocognitive changes in fluid and crystallized age-adjusted cognition using the National Institutes of Health (NIH) toolbox and self and parent-reported total quality of life using the Pediatric Quality of Life Inventory. Results: Twenty-two percent of eligible participants enrolled in the trial. General linear models did not reveal statistically significant differences between groups. Within group analyses using paired t-tests demonstrated improvement in age-adjusted fluid cognition [t (13) = 3.39, p = 0.005, Cohen's d = 0.61] and crystallized cognition [t (13) = 2.63, p = 0.02, Cohen's d = 0.70] within the aerobic training group but no significant improvement within the stretching group. Paired t-tests demonstrated significant improvement in both self-reported and parent-reported total quality of life measures in the aerobic training group [self-report t (13) = 3.51, p = 0.004, Cohen's d = 0.94; parent-report t (13) = 6.5, p < 0.0001, Cohen's d = 1.80] and the stretching group [self-report t (14) = 4.20, p = 0.0009, Cohen's d = 1.08; parent-report t (14) = 4.06, p = 0.0012, Cohen's d = 1.045]. Conclusion: Quality of life improved significantly in both the aerobic exercise and stretching groups; however, this study suggests that only sub-symptom exacerbation aerobic training was potentially beneficial for neurocognitive recovery, particularly the fluid cognition subset in the NIH Toolbox. Limited sample size and variation in outcomes measures limited ability to detect between group differences. Future research should focus on developing larger studies to determine optimal timing post-injury and intensity of active rehabilitation to facilitate neurocognitive recovery and improve quality of life after mTBI. Clinical Trial Registration: www.ClinicalTrials.gov, identifier: NCT02035579.


Lipid analysis by thin-layer chromatography--a review of the current state.
High-performance thin-layer chromatography (HPTLC) is a widely used, fast and relatively inexpensive method of separating complex mixtures. It is particularly useful for smaller, apolar compounds and offers some advantages over HPLC. This review gives an overview about the special features as well as the problems that have to be considered upon the HPTLC analysis of lipids. The term "lipids" is used here in a broad sense and comprises fatty acids and their derivatives as well as substances related biosynthetically or functionally to these compounds. After a short introduction regarding the stationary phases and the methods how lipids can be visualized on an HPTLC plate, the individual lipid classes will be discussed and the most suitable solvent systems for their separation indicated. The focus will be on lipids that are most abundant in biological systems, i.e. cholesterol and its derivates, glycerides, sphingo- and glycolipids as well as phospholipids. Finally, a nowadays very important topic, the combination between HPTLC and mass spectrometric (MS) detection methods will be discussed. It will be shown that this is a very powerful method to investigate the identities of the HPTLC spots in more detail than by the use of common staining methods. Future aspects of HPTLC in the lipid field will be also discussed.


ATP-dependent calcium uptake by rat liver plasma membrane vesicles. Effect of alkylating hepatotoxins in vivo.
Lethal cell injury from hepatotoxic drugs has been postulated to result from an alteration in cell Ca2+ homeostasis. ATP-dependent Ca2+ uptake by the plasma membrane has a sulfhydryl-dependent functional moiety and, therefore, could be vulnerable to chemically reactive drug intermediates. Thus, alkylating hepatotoxins given in vivo were examined for their ability to inhibit Ca2+ accumulation by plasma membrane vesicles isolated from livers of adult male rats. ATP-dependent Ca2+ accumulation was decreased 62% by bromobenzene, 76% by acetaminophen, and 92% by CCl4. Mitochondrial Ca2+ uptake was minimally affected by the toxins, and only CCl4 affected Ca2+ accumulation by liver microsomes. The effect of acetaminophen on plasma membrane Ca2+ uptake was apparent as early as 45 min postdose. Depletion of protective intracellular GSH by diethyl maleate treatment (400 mg/kg) alone minimally decreased control plasma membrane uptake activity, although the GSH depletion markedly potentiated the effect of acetaminophen on the plasma membrane and on necrosis. Alkylation of sites on the plasma membrane may be a key chemical-macromolecule interaction in drug-induced liver necrosis, and inhibition of plasma membrane Ca2+ regulation may provide a connecting link between the alkylation hypothesis and the perturbed Ca2+ homeostasis hypothesis of lethal cell injury.


Prospective study of ampicillin rash. Report of a collaborative study group.
A multicentre prospective study of 933 patients being treated with ampicillin showed a rash incidence of 7.3%. The rash was commonest in women and patients suffering from viral infections. Most rashes were maculopapular and were not associated with features of true penicillin allergy. We conclude that the development of a maculopapular rash during or after treatment with ampicillin is not in itself a contraindication to future treatment with the penicillins.


From evidence to action to deliver a healthy start for the next generation.
Remarkable progress has been made towards halving of maternal deaths and deaths of children aged 1-59 months, although the task is incomplete. Newborn deaths and stillbirths were largely invisible in the Millennium Development Goals, and have continued to fall between maternal and child health efforts, with much slower reduction. This Series and the Every Newborn Action Plan outline mortality goals for newborn babies (ten or fewer per 1000 livebirths) and stillbirths (ten or fewer per 1000 total births) by 2035, aligning with A Promise Renewed target for children and the vision of Every Woman Every Child. To focus political attention and improve performance, goals for newborn babies and stillbirths must be recognised in the post-2015 framework, with corresponding accountability mechanisms. The four previous papers in this Every Newborn Series show the potential for a triple return on investment around the time of birth: averting maternal and newborn deaths and preventing stillbirths. Beyond survival, being counted and optimum nutrition and development is a human right for all children, including those with disabilities. Improved human capital brings economic productivity. Efforts to reach every woman and every newborn baby, close gaps in coverage, and improve equity and quality for antenatal, intrapartum, and postnatal care, especially in the poorest countries and for underserved populations, need urgent attention. We have prioritised what needs to be done differently on the basis of learning from the past decade about what has worked, and what has not. Needed now are four most important shifts: (1) intensification of political attention and leadership; (2) promotion of parent voice, supporting women, families, and communities to speak up for their newborn babies and to challenge social norms that accept these deaths as inevitable; (3) investment for effect on mortality outcome as well as harmonisation of funding; (4) implementation at scale, with particular attention to increasing of health worker numbers and skills with attention to high-quality childbirth care for newborn babies as well as mothers and children; and (5) evaluation, tracking coverage of priority interventions and packages of care with clear accountability to accelerate progress and reach the poorest groups. The Every Newborn Action Plan provides an evidence-based roadmap towards care for every woman, and a healthy start for every newborn baby, with a right to be counted, survive, and thrive wherever they are born.


Integrated Bioinformatics Data Analysis Reveals Prognostic Significance Of SIDT1 In Triple-Negative Breast Cancer.
Triple-negative breast cancer (TNBC) is a heterogeneous disease with a worse prognosis. However, current therapies have rarely improved the outcome of patients with TNBC. Here we sought to identify novel biomarkers or targets for TNBC. Patients GSE76275 clinic traits and their corresponding mRNA profiles for 198 TNBC and 67 non-TNBC were obtained from the GEO database. Weighted gene co-expression network analysis (WGCNA) of the GSE76275 keyed out hub genes, and the differentially expressed genes (DEGs) were identified with the cut-off of adjusted P (adj. P) <0.01 and |log2 fold-change (FC)| > 1.5. The hub - DEGs overlapping genes, as key genes, were considered for further study using Kaplan-Meier plotter online analysis. Subsequently, Breast Cancer Gene-Expression Miner v4.0 and tissue microarray analysis were applied to determine the transcriptional and translational levels of every key gene. Following plasmid transfection for overexpression, the proliferation of TNBC cells was determined by CCK8 and colony formation assay. Moreover, xenograft tumor models were canvassed to investigate their effect upon in vivo tumor growth. Four genes (SIDT1, ANKRD30A, GPR160, and CA12) were found to be associated with relapse-free survival (RFS) in TNBC through WGCNA and DEGs integrated analysis. Patients with a higher level of SIDT1 had significantly better RFS compared to those with lower levels. The transcriptional and translational levels of SIDT1 were validated as downregulated in patients with triple-negative status, negative estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2). Furthermore, SIDT1 inhibited proliferation of breast cancer cells (MDA-MB-231 and MDA-MB-468) and xenograft studies demonstrated that SIDT1 can suppress tumor growth in vivo. This study suggests that SIDT1 may play a crucial role in TNBC progression and has the potential as a prognostic biomarker of TNBC.