The Vitis vinifera C-repeat binding protein 4 (VvCBF4) transcriptional factor enhances freezing tolerance in wine grape.
Chilling and freezing can reduce significantly vine survival and fruit set in Vitis vinifera wine grape. To overcome such production losses, a recently identified grapevine C-repeat binding factor (CBF) gene, VvCBF4, was overexpressed in grape vine cv. 'Freedom' and found to improve freezing survival and reduced freezing-induced electrolyte leakage by up to 2 °C in non-cold-acclimated vines. In addition, overexpression of this transgene caused a reduced growth phenotype similar to that observed for CBF overexpression in Arabidopsis and other species. Both freezing tolerance and reduced growth phenotypes were manifested in a transgene dose-dependent manner. To understand the mechanistic basis of VvCBF4 transgene action, one transgenic line (9-12) was genotyped using microarray-based mRNA expression profiling. Forty-seven and 12 genes were identified in unstressed transgenic shoots with either a >1.5-fold increase or decrease in mRNA abundance, respectively. Comparison of mRNA changes with characterized CBF regulons in woody and herbaceous species revealed partial overlaps, suggesting that CBF-mediated cold acclimation responses are widely conserved. Putative VvCBF4-regulon targets included genes with functions in cell wall structure, lipid metabolism, epicuticular wax formation and stress-responses suggesting that the observed cold tolerance and dwarf phenotypes are the result of a complex network of diverse functional determinants.


Sequential effects modulate spatial biases.
Healthy individuals usually display a bias toward the left side of space. This effect can be measured in a line bisection task or, alternatively, in a landmark task where prebisected lines are presented to participants. Several factors have been shown to influence pseudoneglect, that is, to vary the magnitude of the left side bias. We performed 2 landmark experiments: 1 online (n = 801) and a 2nd in the laboratory (n = 20). Our results demonstrate that pseudoneglect is strongly modulated by the sequence of trials in a landmark task. Of particular relevance is the fact that, for some histories of responses, pseudoneglect is inverted such that apparently there is a preference for the right side. In addition, we show that the way in which the point of subjective equality depends on the previous sequence of trials is well approximated by an exponential filter, well known from the literature of sequential effects to be related to motor control. In other words, the type of sequential effects we encountered in the landmark task is consistent with a purely motor contribution, further deepening our understanding of the way motor control influences pseudoneglect. (PsycINFO Database Record


A novel mouse model for the hyper-IgM syndrome: a spontaneous activation-induced cytidine deaminase mutation leading to complete loss of Ig class switching and reduced somatic hypermutation.
We describe a spontaneously derived mouse line that completely failed to induce Ig class switching in vitro and in vivo. The mice inherited abolished IgG serum titers in a recessive manner caused by a spontaneous G → A transition mutation in codon 112 of the aicda gene, leading to an arginine to histidine replacement (AID(R112H)). Ig class switching was completely reconstituted by expressing wild-type AID. Mice homozygous for AID(R112H) had peripheral B cell hyperplasia and large germinal centers in the absence of Ag challenge. Immunization with SRBCs elicited an Ag-specific IgG1 response in wild-type mice, whereas AID(R112H) mice failed to produce IgG1 and had reduced somatic hypermutation. The phenotype recapitulates the human hyper-IgM (HIGM) syndrome that is caused by point mutations in the orthologous gene in humans, and the AID(R112H) mutation is frequently found in HIGM patients. The AID(R112H) mouse model for HIGM provides a powerful and more precise tool than conventional knockout strategies.


Older People's Perspectives Regarding the Use of Sensor Monitoring in Their Home.
The early detection of a decline in daily functioning of independently living older people can aid health care professionals in providing preventive interventions. To monitor daily activity patterns and, thereby detect a decline in daily functioning, new technologies, such as sensors can be placed in the home environment. The purpose of this qualitative study was to determine the perspectives of older people regarding the use of sensor monitoring in their daily lives. We conducted indepth, semistructured interviews with 11 persons between 68 and 93 years who had a sensor monitoring system installed in their home. The data were analyzed using Interpretative Phenomenological Analysis. The interviewed older persons positively valued sensor monitoring and indicated that the technology served as a strategy to enable independent living. The participants perceived that the system contributed to their sense of safety as an important premise for independent living. Some of the participants stated that it helped them to remain active. The potential privacy violation was not an issue for the participants. The participants considered that health care professionals' continuous access to their sensor data and use of the data for their safety outweighed the privacy concerns. These results provide new evidence that older persons experience sensor monitoring as an opportunity or strategy that contributes to independent living and that does not disturb their natural way of living. Based on this study, the development of new strategies to provide older people with access to their sensor data must be further explored.


Establishing a Regional Glaucoma Physician Collaborative to Improve Quality of Care.
Improving adherence to practice guidelines can improve patient safety and quality of care. We sought to establish a regional glaucoma physician collaborative to evaluate and improve adherence to the American Academy of Ophthalmology's Primary Open-angle Glaucoma (POAG) Preferred Practice Pattern (PPP) guidelines. Prospective interventional study. The collaborative consisted of 13 glaucoma specialists from 3 practices in Michigan. All consecutive POAG new patient visits were reviewed from each study site to determine physician adherence to the 13 major examination elements of the PPP. In phase 1 of the study, physician adherence rates for each of the recommended examination elements were combined and averaged for all groups. Averages for the collaborative were reported to each site, and each physician received his or her individual adherence rates. Physicians discussed strategies to improve overall adherence to the PPP. Adherence rates were collected in phase 2 to determine if feedback and sharing of strategies resulted in improved adherence. A total of 274 new POAG patient visits from phase 1 and 280 visits from phase 2 were reviewed. After accounting for multiple comparisons, overall improvement approached statistical significance for the evaluation of visual function (91.2% to 96.1%, P < .02) and target intraocular pressure determination (73.7% to 83.2%, P < .01). Improvement for other measures that had a high rate of adherence at baseline (eg, ocular history, pupil examination, and central corneal thickness measurement) was not statistically significant. It is feasible to establish a regional glaucoma physician collaborative to improve standardization of care for patients with newly diagnosed POAG.


Sub-cellular targeting of constitutive NOS in health and disease.
Constitutive nitric oxide synthases (NOSs) are ubiquitous enzymes that play a pivotal role in the regulation of myocardial function in health and disease. The discovery of both a neuronal NOS (nNOS) and an endothelial NOS (eNOS) isoform in the myocardium and the availability of genetically modified mice with selective eNOS or nNOS gene deletion have been of crucial importance for understanding the role of constitutive nitric oxide (NO) production in the myocardium. eNOS and nNOS are homologous in structure and utilize the same co-factors and substrates; however, they differ in their subcellular localization, regulation, and downstream signaling, all of which may account for their distinct effects on excitation-contraction coupling. In particular, eNOS-derived NO has been reported to increase left ventricular (LV) compliance, attenuate beta-adrenergic inotropy and enhance parasympathetic/muscarinic responses, and mediate the negative inotropic response to β3 adrenoreceptor stimulation via cGMP-dependent signaling. Conversely, nNOS-derived NO regulates basal myocardial inotropy and relaxation by inhibiting the sarcolemmal Ca(2+) current (I(Ca)) and promoting protein kinase A-dependent phospholamban (PLN) phosphorylation, independent of cGMP. By inhibiting the activity of myocardial oxidase systems, nNOS regulates the redox state of the myocardium and contributes to maintain eNOS "coupled" activity. After myocardial infarction, up-regulation of myocardial nNOS attenuates adverse remodeling and prevents arrhythmias whereas uncoupled eNOS activity in murine models of left ventricular pressure overload accelerates the progress towards heart failure. Here we review the evidence in support of the idea that NOS subcellular localization, mode of activation, and downstream signaling account for the diverse and highly specialized actions of NO in the heart. This article is part of a Special Issue entitled "Local Signaling in Myocytes".


Root Collar Excavation for Postinfection Control of Armillaria Root Disease of Grapevine.
Root collar excavation for control of Armillaria root disease of grapevine was investigated in two California vineyards (vineyard K1 and vineyard N1) from 2002 to 2004. The hypothesis tested was that root collar excavation, when timed in early stages of root collar infection, may cause mycelial fans of the pathogen Armillaria mellea to recede from the root collar before severe disease results from vascular tissue decay. In vineyard N1, excavation significantly increased yield and cluster weight of symptomatic grapevines; symptomatic-excavated grapevines had the same high mean cluster weight as healthy grapevines, and there were no significant effects of excavation on yield or pruning weight of healthy grapevines. In vineyard K1, where excavated root collars frequently refilled with soil, excavation had no significant effects on yield or pruning weight of symptomatic grapevines, and significantly reduced pruning weight and shoot weight of healthy grapevines. Reexamination in March 2004 revealed that mycelial fans had receded from root collars of symptomatic-excavated grapevines, but remained on root collars of symptomatic-nonexcavated grapevines. Root collar excavation appears to be a promising cultural approach for control of Armillaria root disease, as long as excavated root collars are kept clear of soil.


Frequency of Self-Monitoring of Blood Glucose during the School Day Is Associated with the Optimal Glycemic Control among Korean Adolescents with Type 1 Diabetes.
This study aimed to evaluate the relationship between the frequency of self-monitoring of blood glucose (SMBG) and glycosylated hemoglobin (HbA1c) levels among Korean adolescents with type 1 diabetes mellitus (T1DM). Factors affecting the SMBG frequency were analyzed in order to improve their glycemic control. Sixty-one adolescents aged 13 to 18 years with T1DM were included from one tertiary center. Clinical and biochemical variables were recorded. Factors associated with SMBG frequency were assessed using structured self-reported questionnaires. Average total daily SMBG frequency was 3.8±2.1 and frequency during the school day was 1.3±1.2. The mean HbA1c level was 8.6%±1.4%. As the daily SMBG frequency increased, HbA1c levels declined (P=0.001). The adjusted odds of achieving the target HbA1c in participants who performed daily SMBG ≥5 significantly increased 9.87 folds (95% confidence interval [CI], 1.58 to 61.70) compared with those performed SMBG four times a day. In the subjects whose SMBG frequency <1/day during the school day, an 80% reduction in the adjusted odds ratio 0.2 (95% CI, 0.05 to 0.86) showed compared to the group with performing two SMBG measurements in the school setting. The number of SMBG testing performed at school was significantly high for individuals assisted by their friends (P=0.031) and for those who did SMBG in the classrooms (P=0.039). Higher SMBG frequency was significantly associated with lower HbA1c in Korean adolescents with T1DM. It would be necessary to establish the school environments that can facilitate adequate glycemic control, including frequent SMBG.


X-linked adrenoleukodystrophy in a 6-year-old boy initially presenting with psychiatric symptoms.
X-linked adrenoleukodystrophy (ALD) leads to demyelination of the nervous system, adrenal insufficiency and accumulation of long-chain fatty acids. Most young patients with X-linked ALD develop seizures and progressive neurologic deficits, and die within the first two decades of life. We present the case of a 6-year-old with childhood-onset ALD, first presenting with psychiatric symptoms and progressive gait difficulties, slurred speech and cognitive impairment. Genetic testing was performed and a p.R401Q (c.1202G>A) mutation detected in the ABCD1 gene. ALD should be considered in the differential diagnosis of patients presenting with behavior changes and white matter disease in neuroimaging.


Biology and genetic diversity of phasey bean mild yellows virus, a common virus in legumes in Australia.
This study examined the natural and experimental host range and aphid and graft transmission of the tentative polerovirus phasey bean mild yellows virus (PBMYV). Eleven complete coding sequences from PBMYV isolates were determined from a range of hosts and locations. We found two genetically distinct variants of PBMYV. PBMYV-1 was the originally described variant, and PBMYV-2 had a large putative recombination in open reading frame 5 such that PBMYV-1 and PBMYV-2 shared only 65-66% amino acid sequence identity in the P5 protein. The virus was transmitted by a clonal colony of cowpea aphids (Aphis craccivora) and by grafting with infected scions but was not transmitted by a clonal colony of green peach aphids (Myzus persicae). PBMYV was found in natural infections in 11 host species with a range of symptoms and severity, including seven important grain legume crops from across a wide geographic area in Australia. PBMYV was common and widespread in the tropical weed phasey bean (Macroptilium lathyroides), but it is likely that there are other major alternative hosts for the virus in temperate regions of Australia. The experimental host range of PBMYV included the Fabaceae hosts chickpea (Cicer arietinum), faba bean (Vicia faba), pea (Pisum sativum), and phasey bean, but transmissions failed to infect several other members of the families Asteraceae, Cucurbitaceae, Fabaceae and Solanaceae. PBMYV was commonly found in grain legume crops in eastern and western Australia, sometimes at greater than 90% incidence. This new knowledge about PBMYV warrants further assessments of its economic impact on important grain legume crops.


Alzheimer's disease: the strength of association of costs with different measures of disease severity.
To identify Alzheimer's disease (AD) severity measures for use in cost-effectiveness models that effectively capture the impact of AD on costs. A review of the literature and data abstraction from papers that present 1) mean AD costs (direct, indirect, or total) by disease severity, defined using measure of cognition, functional status, and behavior; and/or 2) the results of regression analyses that estimate the strength of the association between AD costs and disease severity. All papers reviewed showed that mean total costs increase with disease severity regardless of severity-measurement method. The relative difference in mean total costs between patients with severe disease compared to those with moderate disease, or moderate disease compared to mild disease, was fairly consistent across studies, suggesting that any of the disease-severity measures may be used to broadly categorize patients by cost. However, when regression analysis included multiple disease-severity measures, independent associations with costs were noted for the different measures. Cognitive and functional status measures were consistently associated with direct costs, whereas functional status and behavioral measures were consistently associated with indirect costs and caregiver hours. Either multidimensional disease-severity measures, or a single disease-severity measure, that capture the impact of cognition, functional status, and behavior on costs are needed for cost-effectiveness models.


p70S6 kinase signals cell survival as well as growth, inactivating the pro-apoptotic molecule BAD.
Cytokines often deliver simultaneous, yet distinct, cell growth and cell survival signals. The 70-kDa ribosomal protein S6 kinase (p70S6K) is known to regulate cell growth by inducing protein synthesis components. We purified membrane-based p70S6K as a kinase responsible for site-specific phosphorylation of BAD, which inactivates this proapoptotic molecule. Rapamycin inhibited mitochondrial-based p70S6K, which prevented phosphorylation of Ser-136 on BAD and blocked cell survival induced by insulin-like growth factor 1 (IGF-1). Moreover, IGF-1-induced phosphorylation of BAD Ser-136 was abolished in p70S6K-deficient cells. Thus, p70S6K is itself a dual pathway kinase, signaling cell survival as well as growth through differential substrates which include mitochondrial BAD and the ribosomal subunit S6, respectively.


Corticosteroid-Binding Globulin Deficiency Specifically Impairs Contextual and Recognition Memory Consolidation in Male Mice.
Glucocorticoids are essential in modulating memory processes of emotionally arousing experiences and we have shown that corticosteroid-binding globulin (CBG) influences glucocorticoid delivery to the brain. Here, we investigated the role of CBG in contextual and recognition long-term memory according to stress intensity. We used adult male mice totally deficient in CBG (Cbg KO) or brain-specific Cbg KO (CbgCamk KO) to examine their performance in contextual fear conditioning (CFC) and au-ditory fear conditioning, both at short (1 h) and long-term (24 h). Long-term memory in Cbg KO was further analyzed in conditioned odor aversion and in novel object recognition task (NORT) with different paradigms, that is, with and without prior habituation to the context, with a mild or strong stressor applied during consolidation. In the NORT experiments, total and free glucocorticoid levels were measured during consolidation. Impaired memory was observed in the Cbg KO but not in the CbgCamk KO in the CFC and the NORT without habituation when tested 24 h later. However, Cbg KO displayed normal behavior in the NORT with previous habituation and in the NORT with a mild stressor. In condition of the NORT with a strong stressor, Cbg KO retained good 24 h memory performance while controls were impaired. Total and free glucocorticoids levels were always higher in controls than in Cbg KO except in NORT with mild stressor where free glucocorticoids were equivalent to controls. These data indicate that circulating but not brain CBG influences contextual and recognition long-term memory in relation with glucocorticoid levels.


Inhibition of hypoxia inducible factor 1 and topoisomerase with acriflavine sensitizes perihilar cholangiocarcinomas to photodynamic therapy.
Photodynamic therapy (PDT) induces tumor cell death by oxidative stress and hypoxia but also survival signaling through activation of hypoxia-inducible factor 1 (HIF-1). Since perihilar cholangiocarcinomas are relatively recalcitrant to PDT, the aims were to (1) determine the expression levels of HIF-1-associated proteins in human perihilar cholangiocarcinomas, (2) investigate the role of HIF-1 in PDT-treated human perihilar cholangiocarcinoma cells, and (3) determine whether HIF-1 inhibition reduces survival signaling and enhances PDT efficacy. Increased expression of VEGF, CD105, CD31/Ki-67, and GLUT-1 was confirmed in human perihilar cholangiocarcinomas. PDT with liposome-delivered zinc phthalocyanine caused HIF-1α stabilization in SK-ChA-1 cells and increased transcription of HIF-1α downstream genes. Acriflavine was taken up by SK-ChA-1 cells and translocated to the nucleus under hypoxic conditions. Importantly, pretreatment of SK-ChA-1 cells with acriflavine enhanced PDT efficacy via inhibition of HIF-1 and topoisomerases I and II. The expression of VEGF, CD105, CD31/Ki-67, and GLUT-1 was determined by immunohistochemistry in human perihilar cholangiocarcinomas. In addition, the response of human perihilar cholangiocarcinoma (SK-ChA-1) cells to PDT with liposome-delivered zinc phthalocyanine was investigated under both normoxic and hypoxic conditions. Acriflavine, a HIF-1α/HIF-1β dimerization inhibitor and a potential dual topoisomerase I/II inhibitor, was evaluated for its adjuvant effect on PDT efficacy. HIF-1, which is activated in human hilar cholangiocarcinomas, contributes to tumor cell survival following PDT in vitro. Combining PDT with acriflavine pretreatment improves PDT efficacy in cultured cells and therefore warrants further preclinical validation for therapy-recalcitrant perihilar cholangiocarcinomas.


Metabolic response of potato plants to an antisense reduction of the P-protein of glycine decarboxylase.
Potato (Solanum tuberosum L. cv. Desiré) plants with reduced amounts of P-protein, one of the subunits of glycine decarboxylase (GDC), have been generated by introduction of an antisense transgene. Two transgenic lines, containing about 60-70% less P-protein in the leaves compared to wild-type potato, were analysed in more detail. The reduction in P-protein amount led to a decrease in the ability of leaf mitochondria to decarboxylate glycine. Photosynthetic and growth rates were reduced but the plants were viable under ambient air and produced tubers. Glycine concentrations within the leaves were elevated up to about 100-fold during illumination. Effects on other amino acids and on sucrose and hexoses were minor. Nearly all of the glycine accumulated during the day was metabolised during the following night. The data suggest that the GDC operates far below substrate saturation under normal conditions thus allowing a flexible and fast response to changes in the environment.


Crystal structure of the BRCT repeat region from the breast cancer-associated protein BRCA1.
The C-terminal BRCT region of BRCA1 is essential for its DNA repair, transcriptional regulation and tumor suppressor functions. Here we determine the crystal structure of the BRCT domain of human BRCA1 at 2.5 A resolution. The domain contains two BRCT repeats that adopt similar structures and are packed together in a head-to-tail arrangement. Cancer-causing missense mutations occur at the interface between the two repeats and destabilize the structure. The manner by which the two BRCT repeats interact in BRCA1 may represent a general mode of interaction between homologous domains within proteins that interact to regulate the cellular response to DNA damage. The structure provides a basis to predict the structural consequences of uncharacterized BRCA1 mutations.


Strawberries decrease atherosclerotic markers in subjects with metabolic syndrome.
Strawberries have been reported to be potent antioxidants and reduce cardiovascular risk factors, such as elevated blood pressure, hyperglycemia, dyslipidemia, and inflammation in limited studies. We hypothesized that freeze-dried strawberry supplementation will improve blood pressure, impaired glucose, dyslipidemia, or circulating adhesion molecules in obese subjects with metabolic syndrome, thereby lowering cardiovascular risk factors in these subjects. Twenty-seven subjects with metabolic syndrome (2 males and 25 females; body mass index, 37.5 +/- 2.15 kg/m(2); age, 47.0 +/- 3.0 years [means +/- SE]) consumed 4 cups of freeze-dried strawberry beverage (50 g freeze-dried strawberries approximately 3 cups fresh strawberries) or equivalent amounts of fluids (controls, 4 cups of water) daily for 8 weeks in a randomized controlled trial. Anthropometrics and blood pressure measurements, assessment of dietary intakes, and fasting blood draws were conducted at screen and 8 weeks of the study. Strawberry supplementation significantly decreased total and low-density lipoprotein cholesterol (5.8 +/- 0.2 to 5.2 +/- 0.2 mmol/L and 3.5 +/- 0.2 to 3.1 +/- 0.1 mmol/L, respectively [means +/- SE], P < .05) and small low-density lipoprotein particles using nuclear magnetic resonance-determined lipoprotein subclass profile vs controls at 8 weeks (794.6 +/- 94.0 to 681.8 +/- 86.0 nmol/L [means +/- SE], P < .05). Strawberry supplementation further decreased circulating levels of vascular cell adhesion molecule-1 vs controls at 8 weeks (272.7 +/- 17.4 to 223.0 +/- 14.0 ng/mL [means +/- SE], P < .05). Serum glucose, triglycerides, high-density lipoprotein cholesterol, blood pressure, and waist circumference were not affected. Thus, short-term freeze-dried strawberry supplementation improved selected atherosclerotic risk factors, including dyslipidemia and circulating adhesion molecules in subjects with metabolic syndrome, and these results need confirmation in future trials.


Growth-promoting effect of bisphenol A on neuroblastoma in vitro and in vivo.
To investigate the effect and mechanism of bisphenol A (BPA), one of the main environmental endocrine disruptors, on the proliferation of human neuroblastoma cells. In vitro, cultured SK-N-SH cells were treated with 17beta-estradiol (E(2); 1 ng/mL), BPA (2 microg/mL) with or without estrogen receptor antagonist ICI182,780 (10(-6) mol/L). Viable cell number, DNA proliferation index, and expression of cyclin-dependent kinase 4 and cyclin D1 were assessed by absorbance reading, flow cytometry, and western blotting, respectively. In vivo, ovariectomized nude mice bearing SK-N-SH tumors were administered by gavage with E(2) (500 microg/kg per day, n = 11), BPA (200 mg/kg per day, n = 10), or vehicle (n = 9) for 18 days. Mice body weight, tumor volume and weight were examined every 3 days. Tumor microvessel density, proliferating cell nuclear antigen and vascular endothelial growth factor expression were evaluated by immunohistochemical staining or western blotting. In vitro, the BPA group had 20% higher number of viable cells, 70% higher proliferation index (both P < .01), and higher expression of cyclin-dependent kinase 4 and cyclin D1 than the nontreated group. In vivo, the BPA group had over 50% higher gross tumor volume, tumor weight, microvessel density, proliferating cell nuclear antigen (P < .05 or .01), and higher vascular endothelial growth factor protein expression than the mock control group. Both in vitro and in vivo BPA effects were comparable with those by E(2). ICI182,780 effectively abolished the promoting effect for both. Bisphenol A can promote the growth of neuroblastoma to a level similar to that of E(2). Estrogen receptor-dependent pathway and angiogenesis may contribute to the underlying mechanisms.


Early Screening Parameters for Dysphagia in Acute Ischemic Stroke.
Dysphagia is a frequent symptom in patients with acute stroke. It is associated with malnutrition, aspiration and mortality. The identification of early screening parameters for dysphagia promptly leading to a professional swallowing examination is therefore of utmost importance. This study aimed to detect early and easily assessable predictors of dysphagia in a large cohort of patients with acute ischemic stroke. Our analysis was based on data from a prospective in-hospital registry. Patients with ischemic stroke were included over the course of 3 years. Patients were scheduled to undergo a clinical swallowing investigation within the first 24 h after hospital admission. Step-wise multivariate logistic regression was used to identify independent predictors of dysphagia in general and of pneumonia in particular. 1,646 patients with ischemic stroke were included. Stroke severity in terms of higher National Institute of Health Stroke Scale (NIHSS) values (p < 0.001), male gender (p = 0.006) and higher age (p < 0.001) independently predicted dysphagia. A receiver operating characteristics analysis revealed an NIHSS cut-off value of 4.5 for optimal differentiation between patients with and without dysphagia (sensitivity 0.77; specificity 0.77). Dysphagia (p < 0.001), male gender (p = 0.002), higher NIHSS scores (p < 0.001) and higher age (p = 0.002) were factors that were independently associated with pneumonia. The NIHSS cut-off value for differentiating between patients with and without pneumonia was 5.5 (sensitivity 0.91; specificity 0.67). Stroke severity in terms of NIHSS is a simple and reliable predictor of dysphagia. Patients with NIHSS values ≥5 should be quickly directed towards a professional swallowing examination. Dysphagia was confirmed to be a strong predictor of pneumonia.


[Mechanisms of action of and resistance to rifampicin and isoniazid in Mycobacterium tuberculosis: new information on old friends].
Human tuberculosis is still one of the most frequent causes of death worldwide. Despite the implementation of therapeutic regimes combining four drugs, the rise of resistant and multidrug-resistant Mycobacterium tuberculosis strains has compromised their efficacy. Two of the most effective anti-tubercular drugs in use, rifampicin and isoniazid, have been closely studied due to their therapeutic importance. These studies have led to the identification of the genes involved in resistance mechanisms and of those encoding the molecular targets for these drugs. Rifampicin is an inhibitor of the beta-subunit of the RNA polymerase of prokaryotes, including M. tuberculosis. Resistance to rifampicin is mediated by mutations clustered in a small region of the rpoB gene. A fraction of resistant strains showed no mutations in rpoB, suggesting that other mechanisms of resistance, possibly efflux pumps, may exist. Isoniazid is a pro-drug activated by KatG, a catalase-peroxidase. Mutations in katG, the most commonly found in M. tuberculosis clinical isolates, give high levels of resistance. In spite of this, the molecular target for isoniazid is InhA, an enoyl-ACP-reductase involved in the biosynthesis of mycolic acids. Other mutations causing resistance to isoniazid have been mapped to ndh, a gene encoding the NADH dehydrogenase.


A Prolonged Response and Characteristics of Trabectedin Treatment of Metastatic Soft Tissue Sarcoma.
Unique features and treatment effects of trabectedin are presented in consideration of soft tissue sarcoma management. A prolonged time on trabectedin through 59 cycles is shown. This is one of the longer reported uses of trabectedin successfully to control disease. Adjunctive cytoreduction options with surgery, radiation or ablation are presented. Future studies would be helpful to investigate treatment holidays, the impact of multi-modality care and assessment of genetics of clonal metastases. This may assist in guiding and selecting patients for priority treatment with trabectedin.


Effects of different antigenic microenvironments on the course of CD8+ T cell responses in vivo.
The influence of microenvironment on the course of CD8 + T cell responses in vivo was investigated by injecting H-2Kb-specific T cells from donor TCR transgenic (TCR-Tg) mice into H-2kb-Tg mice. H-2Kb expression in recipients was either ubiquitous (CBK mice) or restricted to myeloid and erythroid cells (K beta mice). Donor T cells proliferated as extensively and acquired similar surface phenotypes in spleen of both recipient types. Thus, neither the restricted pattern of H-2Kb expression nor the significantly reduced level of H-2Kb expression by myeloid cells in Kbeta recipients affects the ability of the splenic microenvironment to prime T cell proliferation in vivo. However, an unsustained burst of cytolytic activity was generated rapidly in spleen of CBK recipients, whereas relatively little cytolytic activity was generated in K beta spleen. This indicates that effector T cells were not generated efficiently in spleen of Kbeta recipients even though extensive T cell proliferation was taking place in this microenvironment. Furthermore, activated donor T cells dispersed rapidly throughout primary and secondary lymphoid organs of Kbeta recipients, whereas few T cells migrated from spleen in CBK recipients. Consequently, the course of CD8+ T cell responses and the anatomical distribution of activated T cells are profoundly influenced by the nature of the antigenic microenvironment encountered in vivo. We conclude that T cells rapidly proliferate and acquire new tissue-homing characteristics but do not differentiate into cytolytic effector cells at the site of priming when they encounter myeloid cells expressing low levels of antigen in vivo.


Centralized sanctioning and legitimate authority promote cooperation in humans.
Social sanctioning is widely considered a successful strategy to promote cooperation among humans. In situations in which individual and collective interests are at odds, incentives to free-ride induce individuals to refrain from contributing to public goods provision. Experimental evidence from public goods games shows that when endowed with sanctioning powers, conditional cooperators can discipline defectors, thus leading to greater levels of cooperation. However, extant evidence is based on peer punishment institutions, whereas in complex societies, systems of control are often centralized: for instance, we do not sanction our neighbors for driving too fast, the police do. Here we show the effect of centralized sanctioning and legitimate authority on cooperation. We designed an adaptation of the public goods game in which sanctioning power is given to a single monitor, and we experimentally manipulated the process by which the monitor is chosen. To increase the external validity of the study, we conducted lab-in-the-field experiments involving 1,543 Ugandan farmers from 50 producer cooperatives. This research provides evidence of the effectiveness of centralized sanctioning and demonstrates the causal effect of legitimacy on cooperation: participants are more responsive to the authority of an elected monitor than a randomly chosen monitor. Our essay contributes to the literature on the evolution of cooperation by introducing the idea of role differentiation. In complex societies, cooperative behavior is not only sustained by mechanisms of selection and reciprocity among peers, but also by the legitimacy that certain actors derive from their position in the social hierarchy.


Spatiotemporal molecular analysis of cyanobacteria blooms reveals Microcystis--Aphanizomenon interactions.
Spatial and temporal variability in cyanobacterial community composition (CCC) within and between eutrophic lakes is not well-described using culture independent molecular methods. We analyzed CCC across twelve locations in four eutrophic lakes and within-lake locations in the Yahara Watershed, WI, on a weekly basis, for 5 months. Taxa were discriminated by length of MspI-digested cpcB/A intergenic spacer gene sequences and identified by comparison to a PCR-based clone library. CCC across all stations was spatially segregated by depth of sampling locations (ANOSIM R = 0.23, p < 0.001). Accordingly, CCC was correlated with thermal stratification, nitrate and soluble reactive phosphorus (SRP, R = 0.2-0.3). Spatial variability in CCC and temporal trends in taxa abundances were rarely correlative between sampling locations in the same lake indicating significant within lake spatiotemporal heterogeneity. Across all stations, a total of 37 bloom events were observed based on distinct increases in phycocyanin. Out of 97 taxa, a single Microcystis, and two different Aphanizomenon taxa were the dominant cyanobacteria detected during bloom events. The Microcystis and Aphanizomenon taxa rarely bloomed together and were significantly anti-correlated with each other at 9 of 12 stations with Pearson R values of -0.6 to -0.9 (p < 0.001). Of all environmental variables measured, nutrients, especially nitrate were significantly greater during periods of Aphanizomenon dominance while the nitrate+nitrite:SRP ratio was lower. This study shows significant spatial variability in CCC within and between lakes structured by depth of the sampling location. Furthermore, our study reveals specific genotypes involved in bloom formation. More in-depth characterization of these genotypes should lead to a better understanding of factors promoting bloom events in these lakes and more reliable bloom prediction models.


Symptom reduction in depression after treatment with L-tryptophan or imipramine. Item analysis of Hamilton rating scale for depression.
Thirty-eight in-patients with endogenous- and 20 in-patients with non-endogenous depression, took part in a multi-centre 3-week double-blind trial where patients were randomly allocated to treatment with either 6 g L-tryptophan or 150 mg imipramine daily. Item analysis of Hamilton ratings, before the investigation and weekly during the trial period demonstrated few statistically different mean scores on individual items between the two treatment groups. After 3 weeks' treatment a statistically significant item mean reduction on the 0.1% level was found in the item Agitation in favour of imipramine-treated, and in the item Work and Activities in favour of L-tryptophan-treated endogenously depressed patients. After 3 weeks' treatment a statistically significant item mean reduction on the 5% level was found in the item Suicide in favour of imipramine-treated non-endogenously depressed patients. The present study has shown that, after 3 weeks' treatment, imipramine and L-tryptophan has decreased the mean score on individual items of HRS in about the same degree.


Comparison of Full Outline of UnResponsiveness (FOUR) score and the conventional scores in predicting outcome in aneurysmal subarachnoid haemorrhage patients.
Full Outline of UnResponsiveness (FOUR) score is a more comprehensive score used to assess eye response, motor response, brainstem reflexes, and respiration that was introduced to overcome the drawbacks of Glasgow coma scale (GCS) score. Our aim was to assess which score best predicts mortality and poor outcome in aneurysmal subarachnoid haemorrhage (aSAH) patients. This cohort study, prospectively evaluated the use of FOUR score to assess the mortality and outcome in aSAH patients during the period from November 2015 to November 2016. For each patient of aSAH, GCS, FOUR score, Hunt and Hess (HH) score and World Federation of Neurological Surgeons (WFNS) score were determined at the time of admission to neurosurgical intensive care unit. All patients were followed till 28 days post-SAH and their outcome were assessed by Glasgow outcome scale (GOS). We calculated the sensitivity (Sn) and specificity (Sp) for each of these scores. We generated the receiver operating characteristic curve (ROC), quantified the accuracy by the area under curve (AUC), and also calculated their 95% confidence interval (95% CI). A total of 75 aSAH patients were enrolled for the study. The mortality was 24/75 (32%) with 23 in-hospital deaths. FOUR score was highly specific (86.27%) and sensitive (75%) for the prediction of mortality. However, for predicting 28-day outcome, WFNS and HH grade were most specific (92.5%), whereas FOUR and HH score was moderately specific (68.57%). FOUR score is among the most specific and moderately sensitive tool for prediction of mortality. However, WFNS and HH grade are more specific in predicting the 28-day outcome.


Structural organization of actin in the sea urchin egg cortex: microvillar elongation in the absence of actin filament bundle formation.
We have investigated the relationship between the formation of actin filament bundles and the elongation of microvilli (MV) after fertilization in sea urchin eggs. In a previous study (1979, J Cell Biol. 83:241-248) we demonstrated that increased pH induced the formation of actin filaments in isolated sea urchin egg cortices with the concomitant elongation of MV. On the basis of these results we suggested that increased cytoplasmic pH after fertilization causes a reorganization of cortical actin, which in turn provides the force for MV elongation. To test this hypothesis, we compared the morphology of microvilli in eggs activated with and without the release of fertilization acid. Activation of eggs in normal sea water with the calcium ionophore A23187 causes the release of fertilization acid and the elongation of MV containing core bundles of actin filaments. Eggs activated with A23187 in NA(+)-free water do not undergo normal fertilization acid release but develop elongated, flaccid MV. These MV contain an irregular network of actin filaments rather than the parallel bundles of filaments found in normal MV. The addition of 40 mM NaCl to these eggs results in the release of H(+) and the concomitant conversion of flaccid MV to erect MV containing typical core bundles of actin filaments. Identical results are obtained when 10 mM NH(4)Cl is substituted for NaCl. The induction of cytoplasmic alkalinization in unactivated eggs with NH(4)Cl does not cause either MV elongation or the formation of actin filament bundles . These results suggest that: (a) the elongation of MV is stimulated by a rise in intracellular free Ca(++) concentration; (b) actin filament bundle formation is triggered by an increase in cytoplasmic pH; and (c) the formation of actin filament bundles is not necessary for MV elongation but is required to provide rigid support for MV.


Three-color multiplex CARS for fast imaging and microspectroscopy in the entire CHn stretching vibrational region.
We present a three-color multiplex coherent anti-Stokes Raman scattering (CARS) setup that facilitates a prompt recording of broadband CARS spectra along with a fast CARS imaging. With separate narrowband Stokes and probe beams being introduced in the near IR, we are able to incorporate a stable, wideband Ti:sapphire femtosecond laser as a pump beam that covers the full range of Raman shift for CHn stretching vibrational modes. Experimentally, high-resolution multiplex CARS signals are allowed to investigate molecular vibrations over the range of 2650 cm-1 - 3050 cm-1, which are spectrally integrated to construct lipid-sensitive images. It is demonstrated that the proposed implementation promises a particular benefit on CARS imaging of lipid-rich tissue structures by providing detailed information on CHn Raman-active vibrations at points of interest on the CARS images that can be obtained at high frame rates.


Primary structure, neural-specific expression, and chromosomal localization of Cux-2, a second murine homeobox gene related to Drosophila cut.
The cut locus of Drosophila encodes a diverged homeodomain-containing protein that is required for the development of external sensory (es) organs and other tissues. A homologous gene (Cux-1) that encodes a transcriptional repressor has been identified in the mouse and other mammals. We have identified a second murine homeobox-containing gene (designated Cux-2) that is structurally related to Drosophila cut. The murine Cux-2 homeobox was similar to Drosophila cut and encoded a homeodomain that contained a characteristic histidine residue at position 50. The predicted Cux-2 protein contained 1426 amino acids and included three internal 60-amino acid repeats (Cut repeats) that were previously found in Drosophila Cut and murine Cux-1. Unlike Cux-1, expression of Cux-2 was restricted to neural tissue. In the adult brain, Cux-2 was prominently expressed in neurons in the thalamus and limbic system. In embryos, Cux-2 was expressed in the developing central and peripheral nervous systems, including the telencephalon and peripheral ganglia of the trigeminal and glossopharyngeal nerves. A glutathione S-transferase fusion protein containing the carboxyl-terminal Cut repeat and homeodomain of Cux-2 exhibited sequence-specific binding to oligonucleotides derived from the promoter of the Ncam gene. Using an interspecific backcross panel, Cux-1 and Cux-2 were mapped to distinct loci that were genetically linked on distal chromosome 5. These results demonstrate that a family of homeobox genes related to Drosophila cut is located on chromosome 5 in the mouse. Cux-2 is expressed exclusively in the central and peripheral nervous systems, and the Cux-2 gene product binds to DNA in a sequence-specific manner. Cux-2 may encode a transcription factor that is involved in neural specification in mammals.


Terminally ill patients' and lay-carers' perceptions and experiences of community-based services.
This paper reports on terminally ill patients' and their lay-carers' perceptions and experiences of community-based services. It forms part of a larger investigation which examined the extent of service provision and any perceived gaps and problems from the perspectives of both clients and providers of community-based assistance for the terminally ill. The participants were nine terminally ill patients and 12 lay-carers receiving community nursing care. Fifteen face-to-face, semi-structured interviews were conducted to elicit their experiences and perceptions of the care being provided. Qualitative, thematic analysis was carried out on the audio-taped interviews. On the whole, respondents reported a high level of satisfaction and appreciation for the help provided. Several areas appeared more problematic, however, and warrant further consideration. These include perceptions of health professionals, particularly their role domains, power and expertise, and some of the practical disadvantages and problems faced by some patients and their families. These are illustrated and the implications for dying patients, their lay-carers and community-based health professionals are discussed.


Sex Differences in 1-Year Outcomes After Percutaneous Coronary Intervention in the Veterans Health Administration.
Advancements in percutaneous coronary intervention (PCI) for treating obstructive coronary artery disease have reduced major adverse events, including mortality. Yet, evidence as to whether women and men experience similar outcomes is mixed. The objective was to examine sex differences in 1-year major adverse cardiac outcomes for the national population of patients undergoing PCI at Veterans Health Administration (VA) cardiac catheterization laboratories. All Veterans undergoing PCI at VA hospitals between October 1, 2007 and September 30, 2013 (N = 64,757; Women = 1,040) were included. Cox proportional hazards models compared 1-year postprocedural outcomes [rehospitalization for myocardial infarction (MI), all-cause mortality, and major adverse cardiovascular events (MACE)] by sex. Women Veterans undergoing PCI were more likely to be younger, black, obese, and have chronic depression and less likely to have common cardiovascular risk factors and to have had prior cardiac events than Veteran men. One-year rates for women versus men were 2.1% and 2.5% for rehospitalization (p-value = 0.57); 3.5% and 4.9% for mortality (p-value = 0.14), and 5.4% and 6.9% for MACE (p-value = 0.18). There were no significant sex differences in any of the outcomes in Cox proportional hazards models. Despite differences in clinical risk factors at the time of PCI, women and men Veterans treated at VA cardiac catheterization laboratories experienced comparable 1-year rehospitalization for MI, mortality, and MACE post-PCI. These results demonstrated similar 1-year post-PCI outcomes for men and women in a national population of patients who have more comorbidities and mental health issues than the general population.


NEXAFS and XPS studies of nitrosyl chloride.
The electronic structure of nitrosyl chloride (ClNO) has been investigated in the gas phase by X-ray Photoelectron (XPS) and Near Edge X-ray Absorption Fine Structure (NEXAFS) spectroscopy at the Cl 2p, Cl 2s, N 1s and O 1s edges in a combined experimental and theoretical study. The theoretical calculations at different levels of approximation predict ionization potential values in good agreement with the experimental data and allow us to assign the main features of the absorption spectra. An unexpected failure of the density functional model is, however, observed in the calculation of the Cl 2s binding energy, which is related to a large self-interaction error. Largely different photoabsorption cross-section patterns are experimentally observed in core excitations from the investigated quantum shells (n = 1, 2). This finding is confirmed by the oscillator strength distributions calculated at different absorption edges; in the case of the n = 2 shell the bands below the threshold are extremely weak and most of the absorption intensity is due to excitations in the continuum.


Do increases in payments for obstetrical deliveries affect prenatal care?
Raising fees is one of the primary means that State Medicaid Programs employ to maintain provider participation. While a number of studies have sought to quantify the extent to which this policy retains or attracts providers, few have looked at the impact of these incentives on patients. In this study, the authors used Medicaid claims data to examine changes in volume and site of prenatal care among women who delivered babies after the Maryland Medicaid Program raised physicians fees for deliveries 200 percent at the end of its 1986 fiscal year. Although the State's intent was to stabilize the pool of nonhospital providers who were willing to deliver Medicaid babies, it was also hoped that women would benefit through greater access to prenatal care, especially care rendered in a nonhospital setting. The authors' hypotheses were that (a) the fee increase for obstetrical deliveries would result in an increase in prenatal visits by women on Medicaid, and (b) the fee increase would lead to a shift in prenatal visits from hospital to community based providers. The data for Maryland's Medicaid claims for the fiscal years 1985 through 1987 were used. Comparisons were made in the average number of prenatal visits and the ratio of hospital to nonhospital prenatal visits before and after the fee increase. Data for continuously enrolled women who delivered in the last 4 months of each fiscal year were analyzed for between and within year differences using Student's t-test and ANOVA techniques. The findings indicate very little overall change in either the amount or location of prenatal care during the year after the large fee increase for deliveries.Though significant increases in the number of prenatal visits occurred for women who lived outside of Baltimore City, it is difficult to attribute these changes solely to the fee increase. Where an effect was observed, it appeared to be greatest in non urban areas of the State, probably because coordination of care by fewer Medicaid providers is more common in such areas.The findings do not support the hypotheses that raising fees for obstetrical deliveries uniformly increase community-based prenatal care. Instead, the findings suggest that tying fee increases for obstetrical deliveries to the amount of prenatal care provided for each patient may be the best way of increasing the commitment of Medicaid obstetrical providers to give their patients more comprehensive perinatal care.


Least-cost pathway models indicate northern human dispersal from Sunda to Sahul.
Archaeological records from Australia provide the earliest, indirect evidence for maritime crossings by early modern humans, as the islands to the north-west of the continent (Wallacea) have never been connected to the mainland. Suggested in 1977 by Joseph B. Birdsell, the two main routes from Sunda (mainland Southeast Asia) to Sahul (Australia-New Guinea), still in debate today, are a northern route through Sulawesi with a landing in New Guinea, or a southern route through Bali, Timor and thence landing in northern Australia. Here we construct least-cost pathway models of human dispersal from Sunda to Sahul at 65 ka and 70 ka by extending previous out-of-Africa least-cost models through the digitization of these routes. We recover overwhelming support for a northern route into Sahul, with a landing location on present-day Misool Island. Minimal support is also recovered for the southern route at 70 ka, with a possible crossing to Sahul from eastern Timor. Review of archaeological records on the Wallacean islands crossed by our northern route indicate a dearth of archaeological research in this region. Meanwhile, the comparatively better studied southern islands still lack any archaeological dates comparable to those known for initial occupation in Sunda and Sahul. Based on our model results we suggest Misool Island as the initial landing site for early modern humans on Sahul and recommend a future focus on archaeological fieldwork in the northern Wallacean islands.


[Psychiatric case management. Chronic Disease Management application experience in a public Mental Health Service].
Aim of this study is to investigate the possible effectiveness of a specific program management needs of patients at high impact health care, case management (CM). The welfare impact is evaluated in terms of the severity of the presented disorder or to other characteristic factors of the individual patient, such as: adherence to the proposed treatments, possible resistance to drug treatment, cognitive structure, the presence of comorbid medical pathologies, abuse/addiction and, more generally, all bio-psycho-social functioning variables that can complicate the treatment of the patient. Twenty five outpatients with chronic schizophrenia (age mean 49,5 yrs) were evaluated through the Camberwell Assessment of Need (CAN20) and Life Skill Profile (LSP) before and after 1 year of CM treatment. General psychopathology was assessed by the Clinical Global Impression (CGI) and the Brief Psychiatric Rating Scale (BPRS). Demographic data were collected, as well as data related to the severity of the disorder: number of hospitalizations and number of switch in drug treatment in the year before the study. Between T0 and T1 there is a significant improvement on CGI-G, BPRS (total and HOST factor), LSP and CAN TOT in patients treated with CM. Moreover, in CM treated patients a 58% reduction of hospitalizations is noted in the year of study. There is a possible effectiveness of CM in improving patient's clinical and social needs in chronic psychiatric diseases. The CM reduces the number of hospitalizations.


Mapping the energy and diffusion landscapes of membrane proteins at the cell surface using high-density single-molecule imaging and Bayesian inference: application to the multiscale dynamics of glycine receptors in the neuronal membrane.
Protein mobility is conventionally analyzed in terms of an effective diffusion. Yet, this description often fails to properly distinguish and evaluate the physical parameters (such as the membrane friction) and the biochemical interactions governing the motion. Here, we present a method combining high-density single-molecule imaging and statistical inference to separately map the diffusion and energy landscapes of membrane proteins across the cell surface at ~100 nm resolution (with acquisition of a few minutes). Upon applying these analytical tools to glycine neurotransmitter receptors at inhibitory synapses, we find that gephyrin scaffolds act as shallow energy traps (~3 kBT) for glycine neurotransmitter receptors, with a depth modulated by the biochemical properties of the receptor-gephyrin interaction loop. In turn, the inferred maps can be used to simulate the dynamics of proteins in the membrane, from the level of individual receptors to that of the population, and thereby, to model the stochastic fluctuations of physiological parameters (such as the number of receptors at synapses). Overall, our approach provides a powerful and comprehensive framework with which to analyze biochemical interactions in living cells and to decipher the multiscale dynamics of biomolecules in complex cellular environments.


An unusual case with membranous lipodystrophy in a hypertensive patient with transepidermal elimination.
Membranous lipodystrophy represents a peculiar type of fat necrosis that is present in patients with various types of skin disease. It is characterized by the presence of microcysts and macrocysts and is lined by amorphous eosinophilic material with a crenelated arabesque appearance. These findings have been associated with lupus erythematosus, diabetes mellitus, erythema nodosum, trauma, etc. We report a case of a 43-year-old woman who had a red to purple asymptomatic indurated plaque, approximately seven cm in diameter and on the left arm. She was a chronic hepatitis B antigen carrier and had hypertension for four years. Histopathology of the biopsied lesion showed transepidermal elimination of altered collagen and elastic fibers, as well as membranous lipodystrophy changes. There were hypertensive vascular changes including lymphohistiocytic infiltration around the vascular wall, swelling of endothelial cells, increased thickness of the vascular walls, and narrowing of the lumen. We report a case showing transepidermal elimination with membranous lipodystrophy. We carefully suggest that the secondary phenomenon of transepidermal elimination was associated with membranous lipodystrophy and degenerate connective tissues.


Are malaria transmission-blocking vaccines acceptable to high burden communities? Results from a mixed methods study in Bo, Sierra Leone.
Malaria transmission-blocking vaccines (TBVs) could help break the cycle of malaria transmission by conferring community rather than individual protection. When introducing new intervention strategies, uptake is dependent on acceptability, not just efficacy. In this exploratory study on acceptability of TBVs in Sierra Leone, it was hypothesized that TBVs would be largely acceptable to adults and health workers in areas with relatively few ongoing malaria interventions, and that (i) knowledge of malaria and vaccines, (ii) health behaviours associated with malaria and vaccines, and (iii) attitudes towards different vaccines types could lead to greater TBV acceptability. This study used a mixed methods approach in Bo, Sierra Leone, to understand community knowledge, attitudes, and practices related to malaria and vaccination in general. This included: (i) a population-based cross-sectional survey (n=615 adults), (ii) 6 focus group discussions with parents, and (iii) 20 key informant interviews. The concept of a TBV was explained to participants before they were asked about their willingness to accept this vaccine modality as part of an integrated malaria elimination programme. This study found that most adults would be willing to receive a TBV vaccine. Respondents noted mostly positive past experiences with adult and childhood vaccinations for other infectious diseases and high levels of engagement in other malaria prevention behaviors such as bed nets. Perceived barriers to TBV acceptance were largely focused on general community-level distribution of a vaccine, including personal fears of vaccination and possible costs. After an explanation of the TBV mechanism, nearly all focus group and interview participants believed that community members would accept the vaccine as part of an integrated malaria control approach. Both parents and health workers offered insight on how to successfully roll-out a future TBV vaccination programme. The willingness of community members in Bo, Sierra Leone to accept a TBV as part of an integrated anti-malarial strategy suggests that the atypical mechanism of TBV action might not be an obstacle to future clinical trials. This study's findings suggests that perceived general barriers to vaccination implementation, such as perceived personal fears and vaccine cost, must be addressed in future clinical and implementation research studies.


Structural basis of a histone H3 lysine 4 demethylase required for stem elongation in rice.
Histone lysine methylation is an important epigenetic modification in regulating chromatin structure and gene expression. Histone H3 lysine 4 methylation (H3K4me), which can be in a mono-, di-, or trimethylated state, has been shown to play an important role in gene expression involved in plant developmental control and stress adaptation. However, the resetting mechanism of this epigenetic modification is not yet fully understood. In this work, we identified a JmjC domain-containing protein, JMJ703, as a histone lysine demethylase that specifically reverses all three forms of H3K4me in rice. Loss-of-function mutation of the gene affected stem elongation and plant growth, which may be related to increased expression of cytokinin oxidase genes in the mutant. Analysis of crystal structure of the catalytic core domain (c-JMJ703) of the protein revealed a general structural similarity with mammalian and yeast JMJD2 proteins that are H3K9 and H3K36 demethylases. However, several specific features were observed in the structure of c-JMJ703. Key residues that interact with cofactors Fe(II) and N-oxalylglycine and the methylated H3K4 substrate peptide were identified and were shown to be essential for the demethylase activity in vivo. Several key residues are specifically conserved in known H3K4 demethylases, suggesting that they may be involved in the specificity for H3K4 demethylation.


Adult human mesenchymal stem cells delivered via intra-articular injection to the knee following partial medial meniscectomy: a randomized, double-blind, controlled study.
There are limited treatment options for tissue restoration and the prevention of degenerative changes in the knee. Stem cells have been a focus of intense preclinical research into tissue regeneration but limited clinical investigation. In a randomized, double-blind, controlled study, the safety of the intra-articular injection of human mesenchymal stem cells into the knee, the ability of mesenchymal stem cells to promote meniscus regeneration following partial meniscectomy, and the effects of mesenchymal stem cells on osteoarthritic changes in the knee were investigated. A total of fifty-five patients at seven institutions underwent a partial medial meniscectomy. A single superolateral knee injection was given within seven to ten days after the meniscectomy. Patients were randomized to one of three treatment groups: Group A, in which patients received an injection of 50 × 10⁶ allogeneic mesenchymal stem cells; Group B, 150 × 10⁶ allogeneic mesenchymal stem cells; and the control group, a sodium hyaluronate (hyaluronic acid/hyaluronan) vehicle control. Patients were followed to evaluate safety, meniscus regeneration, the overall condition of the knee joint, and clinical outcomes at intervals through two years. Evaluations included sequential magnetic resonance imaging (MRI). No ectopic tissue formation or clinically important safety issues were identified. There was significantly increased meniscal volume (defined a priori as a 15% threshold) determined by quantitative MRI in 24% of patients in Group A and 6% in Group B at twelve months post meniscectomy (p = 0.022). No patients in the control group met the 15% threshold for increased meniscal volume. Patients with osteoarthritic changes who received mesenchymal stem cells experienced a significant reduction in pain compared with those who received the control, on the basis of visual analog scale assessments. There was evidence of meniscus regeneration and improvement in knee pain following treatment with allogeneic human mesenchymal stem cells. These results support the study of human mesenchymal stem cells for the apparent knee-tissue regeneration and protective effects.


Host-Cell Survival and Death During Chlamydia Infection.
Different Chlamydia trachomatis strains are responsible for prevalent bacterial sexually-transmitted disease and represent the leading cause of preventable blindness worldwide. Factors that predispose individuals to disease and mechanisms by which chlamydiae cause inflammation and tissue damage remain unclear. Results from recent studies indicate that prolonged survival and subsequent death of infected cells and their effect on immune effector cells during chlamydial infection may be important in determining the outcome. Survival of infected cells is favored at early times of infection through inhibition of the mitochondrial pathway of apoptosis. Death at later times displays features of both apoptosis and necrosis, but pro-apoptotic caspases are not involved. Most studies on chlamydial modulation of host-cell death until now have been performed in cell lines. The consequences for pathogenesis and the immune response will require animal models of chlamydial infection, preferably mice with targeted deletions of genes that play a role in cell survival and death.


Spirometric abnormalities following treatment for pulmonary tuberculosis in Ilorin, Nigeria.
Pulmonary tuberculosis (PTB) contributes significantly to morbidity and mortality worldwide, and despite microbiological cure for the disease, many patients still demonstrate residual respiratory symptoms and spirometric abnormalities. The study aimed at identifying the prevalence, pattern and factors associated with spirometric abnormalities in patients successfully treated for PTB in Ilorin, Nigeria. This was a hospital-based cross-sectional study at the pulmonary outpatient clinics of the University of Ilorin Teaching Hospital and Kwara State Specialist Hospital, Sobi, Ilorin. A total of 308 consenting patients who had been certified microbiologically cured for bacteriologically confirmed PTB in the preceding 3 years had assessment of residual pulmonary symptoms, spirometry and plain chest radiograph. The prevalence of abnormal spirometry following treatment for PTB was 72.1% (confidence interval: 0.6682-0.7695), with restrictive pattern being the predominant abnormality (42.2%). Over half of the patients (56.5%) had at least one residual respiratory symptom. The significant predictors of abnormal spirometry were PTB retreatment (adjusted odds ratio [aOR] = 6.918; P = 0.012), increasing modified Medical Research Council dyspnoea scores (aOR = 7.935; P = 0.008) and increasing radiologic scores (aOR = 4.679; P ≤ 0.001) after treatment. There is significant residual lung function impairment in majority of the individuals successfully treated for PTB in Ilorin. This highlights the need for spirometric assessment and follow-up after treatment.


Perspectives on family caregiving of people aging with intellectual disability affected by dementia: Commentary from the International Summit on Intellectual Disability and Dementia.
This article, an output of the 2016 International Summit on Intellectual Disability and Dementia, examines familial caregiving situations within the context of a support-staging model for adults with intellectual disability (ID) affected by dementia. Seven narratives offer context to this support-staging model to interpret situations experienced by caregivers. The multidimensional model has two fundamental aspects: identifying the role and nature of caregiving as either primary (direct) or secondary (supportive); and defining how caregiving is influenced by stage of dementia. We propose staging can affect caregiving via different expressions: (1) the "diagnostic phase," (2) the "explorative phase," (3) the "adaptive phase," and (4) the "closure phase." The international narratives illustrate direct and indirect caregiving with commonality being extent of caregiver involvement and attention to the needs of an adult with ID. We conclude that the model is the first to empirically formalize the variability of caregiving within families of people with ID that is distinct from other caregiving groups, and that many of these caregivers have idiosyncratic needs. A support-staging model that recognizes the changing roles and demands of carers of people with ID and dementia can be useful in constructing research, defining family-based support services, and setting public policy.


Neuroprotection by Human Dental Pulp Mesenchymal Stem Cells: From Billions to Nano.
Mesenchymal Stem Cell (MSC) therapy in recent years has gained significant attention. Though the functional outcomes following MSC therapy for neurodegenerative diseases are convincing, various mechanisms for the functional recovery are being debated. Nevertheless, recent studies convincingly demonstrated that recovery following MSC therapy could be reiterated with MSC secretome per se thereby shifting the dogma from cell therapy to cell "based" therapy. In addition to various functional proteins, stem cell secretome also includes extracellular membrane vesicles like exosomes. Exosomes which are of "Nano" size have attracted significant interest as they can pass through the bloodbrain barrier far easily than macro size cells or growth factors. Exosomes act as a cargo between cells to bring about significant alterations in target cells. As the importance of exosomes is getting unveil, it is imperial to carry out a comprehensive study to evaluate the neuroprotective potential of exosomes as compared to conventional co-culture or total condition medium treatments. Thus, the present study is designed to compare the neuroprotective potential of MSC derived exosomes with MSC-condition medium or neuron-MSC-co-culture system against kainic acid induced excitotoxicity in in vitro condition. The study also aims at comparing the neuroprotective efficacy of exosomes/condition medium/co-culture of two MSC viz., neural crest derived human Dental Pulp Stem Cells (hDPSC) and human Bone-Marrow Mesenchymal Stem Cells (hBM-MSC) to identify the appropriate MSC source for treating neurodegenerative diseases. Our results demonstrated that neuroprotective efficacy of MSC-exosomes is as efficient as MSC-condition medium or neuron-MSC co-culture system and treating degenerating hippocampal neurons with all three MSC based approaches could up-regulate host's endogenous growth factor expressions and prevent apoptosis by activating cell survival PI3K-B-cell lymphoma-2 (Bcl-2) pathway. Thus, the current study highlights the possibilities of treating neurodegenerative diseases with "Nano" size exosomes as opposed to transplanting billions of stem cells which inherit several disadvantages.


Forensic Medicine and the Military Population: International Dental Records and Personal Identification Concerns.
The first goal of this research was to perceive the global commitment towards the organization and archiving of dental records and to compare it with each country's security risk rating. The second one was to study dental records in a sample of the Portuguese military population, using the available national dental records. An e-mail was sent to representative dentistry associations in several countries, requesting some information concerning the professionals' awareness of this issue. After obtaining permission from the Ethics Committee, the information was collected through the Forensic Dental Symbols® system into the Dental Encoder®, as an extension of a Spanish study, and a generic codification was used (unrestored, restored, missing and crowned teeth). The most common dental record retention period is ten years after treatment. Observing the samples' dental records (595 files), we found a total of 19 040 analyzed teeth, with the following frequencies: unrestored (89.6%), restored (7.0%), missing (2.2%) and crowned (1.1%). There is a wide range of guidelines on how long dentists should keep dental records. Especially for the military population, dental records must include detailed information concerning each tooth situation, in order to support the process of human identification. This article reinforces the need for mandatory quality dental records in all countries, which must be efficiently stored and easily accessible in case dental identification is necessary. For the military population, these requirements are especially important, due to the added risks to which this group is subject.


A declining rate of completion axillary dissection in sentinel lymph node-positive breast cancer patients is associated with the use of a multivariate nomogram.
To compare sentinel lymph node (SLN)-positive breast cancer patients who had completion axillary dissection (ALND) with those who did not, with particular attention to clinicopathologic features, nomogram scores, rates of axillary local recurrence (LR), and changes in treatment pattern over time. While conventional treatment of SLN-positive patients is to perform ALND, there may be a low-risk subgroup of SLN-positive patients in whom ALND is not required. A multivariate nomogram that predicts the likelihood of residual axillary disease may assist in identifying this group. Among 1960 consecutive SLN-positive patients (1997-2004), 1673 (85%) had ALND ("SLN+/ALND") and 287 (15%) did not ("SLN+/no ALND"). We compare in detail the clinicopathologic features, nomogram scores, and rates of axillary LR between groups. Compared with the SLN+/ALND group, patients with SLN+/no ALND were older, had more favorable tumors, were more likely to have breast conservation, had a lower median predicted risk of residual axillary node metastases (9% vs. 37%, P < 0.001), and had a marginally higher rate of axillary LR (2% vs. 0.4%, P = 0.004) at 23 to 30 months' follow-up; half of all axillary LR in SLN+/no ALND patients were coincident with other local or distant sites. For patients in whom intraoperative frozen section was either negative or not done, the rate of completion ALND declined from 79% in 1997 to 62% in 2003 to 2004 but varied widely by surgeon, ranging from 37% to 100%. For 10 of 10 evaluable surgeons, the median nomogram scores in the SLN+/no ALND group were <or=10.5. SLN+/no ALND breast cancer patients, a selected group with relatively favorable disease characteristics, had a 9% predicted likelihood of residual axillary disease by nomogram but an observed axillary LR of 2%. A gradual and significant decline over time in the rate of completion ALND is associated with, but not entirely explained by, the institution of a predictive nomogram. It is reasonable to omit ALND for a low-risk subset of SLN-positive patients.


[Composite matrix membranes as synthetic receptor systems. 2. Structural-morphological characteristics].
Structural and morphological characteristics of composite imprinted membranes for selective recognizing of adenosine 3',5'-cyclic monophosphate (cAMP) were studied. Composite polyvinylidene fluoride microfiltration membranes (Millipore) covered with a thin imprinted polymer layer were prepared using photoinitiated copolymerization of dimethylaminoethylmetacrylate with trimethylolpropanethrimethacrylate in the presence of cAMP as template. Atomic force microscopy (AFM) and scanning electron microscopy (SEM) were used to visualise surfaces and cross-sections of imprinted membranes and to determine their structural and morphological parameters such as pore size, thickness of selective imprinted layer, surface roughness as well as total surface contact area. The impact of structural characteristics on separation properties of the imprinted membranes was analyzed. It was found out that the thickness of the imprinted polymer layer with optimal recognizing properties is limited.


Development and evaluation of an enzyme-linked immunosorbent assay for use in the detection of bovine tuberculosis in cattle.
As a consequence of continued spillover of Mycobacterium bovis into cattle from wildlife reservoirs and increased globalization of cattle trade with associated transmission risks, new approaches such as vaccination and novel testing algorithms are seriously being considered by regulatory agencies for the control of bovine tuberculosis. Serologic tests offer opportunities for identification of M. bovis-infected animals not afforded by current diagnostic techniques. The present study describes assay development and field assessment of a new commercial enzyme-linked immunosorbent assay (ELISA) that detects antibody to M. bovis antigens MPB83 and MPB70 in infected cattle. Pertinent findings include the following: specific antibody responses were detected at ∼90 to 100 days after experimental M. bovis challenge, minimal cross-reactive responses were elicited by infection/sensitization with nontuberculous Mycobacterium spp., and the apparent sensitivity and specificity of the ELISA with naturally infected cattle were 63% and 98%, respectively, with sensitivity improving as disease severity increased. The ELISA also detected infected animals missed by the routine tuberculin skin test, and antibody was detectable in bulk tank milk samples from M. bovis-infected dairy herds. A high-throughput ELISA could be adapted as a movement, border, or slaughter surveillance test, as well as a supplemental test to tuberculin skin testing.


Unity of consciousness.
Although there is much talk in various literatures of streams of consciousness, and most of us have an intuitive understanding of such talk, we are far from having a full grasp of what it is that unifies streams of consciousness, binding together the individual experiences that serve as their constituents. In recent years, discussion of this topic has been principally concerned with synchronic unity of consciousness-the form of unity that is exhibited by momentary states of consciousness, or in other words, by time slices or temporal segments of streams. There are two main questions about synchronic unity. First, what is its scope? Are the simultaneous experiences of a single subject necessarily unified? Generally but not necessarily unified? Sometimes unified? And second, what is the nature of synchronic unity? Is it a fundamental phenomenon, and if not, what are the more basic phenomena that constitute it? This essay reviews recent work on these questions, and provides reasons for preferring some answers to others. This article is categorized under: Philosophy > Consciousness Philosophy > Foundations of Cognitive Science Philosophy > Metaphysics.


Clinical practice guidelines for managing dyslipidemias in kidney transplant patients: a report from the Managing Dyslipidemias in Chronic Kidney Disease Work Group of the National Kidney Foundation Kidney Disease Outcomes Quality Initiative.
The incidence of cardiovascular disease (CVD) is very high in patients with chronic kidney (CKD) disease and in kidney transplant recipients. Indeed, available evidence for these patients suggests that the 10-year cumulative risk of coronary heart disease is at least 20%, or roughly equivalent to the risk seen in patients with previous CVD. Recently, the National Kidney Foundation's Kidney Disease Outcomes Quality Initiative (K/DOQI) published guidelines for the diagnosis and treatment of dyslipidemias in patients with CKD, including transplant patients. It was the conclusion of this Work Group that the National Cholesterol Education Program Guidelines are generally applicable to patients with CKD, but that there are significant differences in the approach and treatment of dyslipidemias in patients with CKD compared with the general population. In the present document we present the guidelines generated by this workgroup as they apply to kidney transplant recipients. Evidence from the general population indicates that treatment of dyslipidemias reduces CVD, and evidence in kidney transplant patients suggests that judicious treatment can be safe and effective in improving dyslipidemias. Dyslipidemias are very common in CKD and in transplant patients. However, until recently there have been no adequately powered, randomized, controlled trials examining the effects of dyslipidemia treatment on CVD in patients with CKD. Since completion of the K/DOQI guidelines on dyslipidemia in CKD, the results of the Assessment of Lescol in Renal Transplantation (ALERT) Study have been presented and published. Based on information from randomized trials conducted in the general population and the single study conducted in kidney transplant patients, these guidelines, which are a modified version of the K/DOQI dyslipidemia guidelines, were developed to aid clinicians in the management of dyslipidemias in kidney transplant patients. These guidelines are divided into four sections. The first section (Introduction) provides the rationale for the guidelines, and describes the target population, scope, intended users, and methods. The second section presents guidelines on the assessment of dyslipidemias (guidelines 1-3), while the third section offers guidelines for the treatment of dyslipidemias (guidelines 4-5). The key guideline statements are supported mainly by data from studies in the general population, but there is an urgent need for additional studies in CKD and in transplant patients. Therefore, the last section outlines recommendations for research.


Ribosome pausing and stacking during translation of a eukaryotic mRNA.
We have devised a sensitive assay to determine the distribution of translating ribosomes on a mRNA. Using this assay to monitor ribosome transit on bovine preprolactin mRNA, we have detected four major positions of ribosome pausing in both wheat-germ and rabbit reticulocyte extracts. Two of these rate-limiting steps represent initiation and termination. One pause occurs after approximately 75 amino acids have been polymerized; signal recognition particle arrests preprolactin synthesis at this position. The other internal pause occurs at 160 amino acids. In these latter two cases ribosomes stop at a GGC glycine codon; however, two other GGC codons are translated without apparent pausing. Surprisingly, we find that up to nine ribosomes are tightly stacked behind each pausing ribosome, such that the ribosome centers are only 27-29 nucleotides apart. The assay should prove useful for probing mechanisms of translational regulation.


Precompetitive Data Sharing as a Catalyst to Address Unmet Needs in Parkinson's Disease.
Parkinson's disease is a complex heterogeneous disorder with urgent need for disease-modifying therapies. Progress in successful therapeutic approaches for PD will require an unprecedented level of collaboration. At a workshop hosted by Parkinson's UK and co-organized by Critical Path Institute's (C-Path) Coalition Against Major Diseases (CAMD) Consortiums, investigators from industry, academia, government and regulatory agencies agreed on the need for sharing of data to enable future success. Government agencies included EMA, FDA, NINDS/NIH and IMI (Innovative Medicines Initiative). Emerging discoveries in new biomarkers and genetic endophenotypes are contributing to our understanding of the underlying pathophysiology of PD. In parallel there is growing recognition that early intervention will be key for successful treatments aimed at disease modification. At present, there is a lack of a comprehensive understanding of disease progression and the many factors that contribute to disease progression heterogeneity. Novel therapeutic targets and trial designs that incorporate existing and new biomarkers to evaluate drug effects independently and in combination are required. The integration of robust clinical data sets is viewed as a powerful approach to hasten medical discovery and therapies, as is being realized across diverse disease conditions employing big data analytics for healthcare. The application of lessons learned from parallel efforts is critical to identify barriers and enable a viable path forward. A roadmap is presented for a regulatory, academic, industry and advocacy driven integrated initiative that aims to facilitate and streamline new drug trials and registrations in Parkinson's disease.


Tumor cell-derived exosomes home to their cells of origin and can be used as Trojan horses to deliver cancer drugs.
Cancer is the second leading cause of death worldwide and patients are in urgent need of therapies that can effectively target cancer with minimal off-target side effects. Exosomes are extracellular nano-shuttles that facilitate intercellular communication between cells and organs. It has been established that tumor-derived exosomes contain a similar protein and lipid composition to that of the cells that secrete them, indicating that exosomes might be uniquely employed as carriers for anti-cancer therapeutics. Methods: We isolated exosomes from two cancer cell lines, then co-cultured each type of cancer cells with these two kinds of exosomes and quantified exosome. HT1080 or Hela exosomes were systemically injected to Nude mice bearing a subcutaneous HT1080 tumor to investigate their cancer-homing behavior. Moreover, cancer cell-derived exosomes were engineered to carry Doxil (a common chemotherapy drug), known as D-exo, were used to detect their target and therapeutic efficacy as anti-cancer drugs. Exosome proteome array analysis were used to reveal the mechanism underly this phenomenon. Results: Exosomes derived from cancer cells fuse preferentially with their parent cancer cells, in vitro. Systemically injected tumor-derived exosomes home to their original tumor tissues. Moreover, compared to Doxil alone, the drug-loaded exosomes showed enhanced therapeutic retention in tumor tissues and eradicated them more effectively in nude mice. Exosome proteome array analysis revealed distinct integrin expression patterns, which might shed light on the underlying mechanisms that explain the exosomal cancer-homing behavior. Conclusion: Here we demonstrate that the exosomes' ability to target the parent cancer is a phenomenon that opens up new ways to devise targeted therapies to deliver anti-tumor drugs.


[Association between mode of delivery and maternal complications in a public hospital in Greater Metropolitan São Paulo, Brazil].
The purpose of this study was to assess the relationship between mode of delivery and maternal complications, based on a retrospective cohort of all births at a public hospital in 2003. Complications included: infection, hemorrhage, hysterectomy, uterine rupture, lesions in adjacent organs, deep venous thrombosis, and pulmonary embolism. The analysis used odds ratio (OR), chi-squared test, and Fisher's exact test, besides logistic regression. Fifteen complications were identified. Taking vaginal delivery as the reference, an association was found between cesarean section and overall complications. Analysis of confounding showed an association between hypertension, HIV, placenta previa, and abruptio placentae. After controlling for these variables, an association remained between overall complications and cesarean section (OR = 9.7; p = 0.04). Another analysis comparing elective cesareans and vaginal deliveries also showed an increased risk for cesarean (OR = 4.7; p = 0.02). Finally, comparing elective cesareans with trial of labor, we found an increased proportion of complications in elective cesareans, with borderline significance (OR = 3; p = 0.058). We concluded that cesarean section is associated with maternal morbidity, even after controlling for confounders.


Comparing the Braden and Jackson/Cubbin Pressure Injury Risk Scales in Trauma-Surgery ICU Patients.
The occurrence of pressure injury in the critical care environment has multiple risk factors. Prevention requires reliable assessment tools to help predict injury risk. The Braden scale, a commonly used risk assessment tool, has been shown to have poor predictive properties in critical care patients. The Jackson/Cubbin scale was developed specifically for pressure injury risk stratification in critically ill patients and has demonstrated acceptable predictive properties in the general critical care population but has not been examined in critically ill trauma-surgical patients. To compare the predictive properties of the Braden and Jackson/Cubbin scales in a trauma-surgical critical care population. A retrospective medical records review was performed to evaluate the clinical characteristics of 366 trauma-surgical critical care patients. Additionally, the negative predictive value, positive predictive value, sensitivity, specificity, and receiver operating characteristic curve with area under the curve of the Braden and Jackson/Cubbin scales were determined. The sample consisted of primarily middle-aged (mean [SD], 56 [19] years) men (64%) admitted after trauma (71%). The participants who developed pressure injuries were older, more often required vasopressors and mechanical ventilation, and were less mobile. Predictive properties for the Braden and Jackson/Cubbin scales, respectively, were as follows: negative predictive value, 78% versus 87%; positive predictive value, 53% versus 66%; sensitivity, 17% versus 54%; specificity, 95% versus 92%; and area under the curve, 0.710 versus 0.793. The Jackson/Cubbin scale demonstrated superior predictive properties and discrimination compared with the Braden scale for pressure injury risk prediction in critically ill trauma-surgical patients.


Central aponeurosis tears of the rectus femoris: practical sonographic prognosis.
This study is a statistical analysis to establish whether a correlation exists between the level and degree of rectus femoris (RF) central tendon injury and the amount of time that an athlete is unable to participate subsequently, referred to as "sports participation absence" (SPA). Causal-comparative study. 35 players from two high-level Spanish soccer teams with an injury to the central tendon of the RF based on clinical and ultrasound criteria. Ultrasound examination was performed with an 8-2 MHz linear multifrequency transducer. All studies included both longitudinal and transverse RF sections. At the proximal level the SPA time is 45.1 days when the injury length is 4.0 cm. This value increases by 5.3 days with each 1 cm increase in the length of injury. In the case of distal level injury, SPA time is 32.9 days when the injury length is 3.9 cm. This value increases by 3.4 days with each 1 cm increase. In the total representative sample, SPA time when the injury length is 4.2 cm corresponds to 39.1 days. This value increases by 4.2 days per length unit. RF central tendon injury at the proximal level is associated with a greater SPA time than at the distal level. Patients with a grade II injury have an SPA time longer than those with a grade I injury whether the injury is located proximal or distal.


Blood mercury concentrations in CHARGE Study children with and without autism.
Some authors have reported higher blood mercury (Hg) levels in persons with autism, relative to unaffected controls. We compared blood total Hg concentrations in children with autism or autism spectrum disorder (AU/ASD) and typically developing (TD) controls in population-based samples, and determined the role of fish consumption in differences observed. The Childhood Autism Risk from Genetics and the Environment (CHARGE) Study enrolled children 2-5 years of age. After diagnostic evaluation, we analyzed three groups: AU/ASD, non-AU/ASD with developmental delay (DD), and population-based TD controls. Mothers were interviewed about household, medical, and dietary exposures. Blood Hg was measured by inductively coupled plasma mass spectrometry. Multiple linear regression analysis was conducted (n = 452) to predict blood Hg from diagnostic status controlling for Hg sources. Fish consumption strongly predicted total Hg concentration. AU/ASD children ate less fish. After adjustment for fish and other Hg sources, blood Hg levels in AU/ASD children were similar to those of TD children (p = 0.75); this was also true among non-fish eaters (p = 0.73). The direct effect of AU/ASD diagnosis on blood Hg not through the indirect pathway of altered fish consumption was a 12% reduction. DD children had lower blood Hg concentrations in all analyses. Dental amalgams in children with gum-chewing or teeth-grinding habits predicted higher levels. After accounting for dietary and other differences in Hg exposures, total Hg in blood was neither elevated nor reduced in CHARGE Study preschoolers with AU/ASD compared with unaffected controls, and resembled those of nationally representative samples.


Smoothelin, a new marker to determine the origin of liver fibrogenic cells.
To explore this hypothesis that smooth muscle cells may be capable of acquiring a myofibroblastic phenotype, we have studied the expression of smoothelin in fibrotic conditions. Normal liver tissue (n = 3) was obtained from macroscopically normal parts of hepatectomy, taken at a distance from hemangiomas. Pathological specimens included post-burn cutaneous hypertrophic scars (n = 3), fibrotic liver tissue (n = 5), cirrhotic tissue (viral and alcoholic hepatitis) (n = 5), and hepatocellular carcinomas (n = 5). Tissue samples were fixed in 10% formalin and embedded in paraffin for immunohistochemistry or were immediately frozen in liquid nitrogen-cooled isopentane for confocal microscopy analysis. Sections were stained with antibodies against smoothelin, which is expressed exclusively by smooth muscle cells, and α-smooth muscle actin, which is expressed by both smooth muscle cells and myofibroblasts. In hypertrophic scars, α-smooth muscle actin was detected in vascular smooth muscle cells and in numerous myofibroblasts present in and around nodules, whereas smoothelin was exclusively expressed in vascular smooth muscle cells. In the normal liver, vascular smooth muscle cells were the only cells that express α-smooth muscle actin and smoothelin. In fibrotic areas of the liver, myofibroblasts expressing α-smooth muscle actin were detected. Myofibroblasts co-expressing α-smooth muscle actin and smoothelin were observed, and their number was slightly increased in parallel with the degree of fibrosis (absent in liver with mild or moderate fibrosis; 5% to 10% positive in liver showing severe fibrosis). In cirrhotic septa, numerous myofibroblasts co-expressed α-smooth muscle actin and smoothelin (more than 50%). In hepatocellular carcinomas, the same pattern of expression for α-smooth muscle actin and smoothelin was observed in the stroma reaction surrounding the tumor and around tumoral cell plates. In all pathological liver samples, α-smooth muscle actin and smoothelin were co-expressed in vascular smooth muscle cells. During development of advanced liver fibrosis, a subpopulation of myofibroblasts expressing smoothelin may be derived from vascular smooth muscle cells, illustrating the different cellular origins of myofibroblasts.


A novel mechanism for regulating hepatic glycogen synthesis involving serotonin and cyclin-dependent kinase-5.
Hepatic autonomic nerves regulate postprandial hepatic glucose uptake, but the signaling pathways remain unknown. We tested the hypothesis that serotonin (5-hydroxytryptamine [5-HT]) exerts stimulatory and inhibitory effects on hepatic glucose disposal. Ligands of diverse 5-HT receptors were used to identify signaling pathway(s) regulating glucose metabolism in hepatocytes. 5-HT had stimulatory and inhibitory effects on glycogen synthesis in hepatocytes mediated by 5-HT1/2A and 5-HT2B receptors, respectively. Agonists of 5-HT1/2A receptors lowered blood glucose and increased hepatic glycogen after oral glucose loading and also stimulated glycogen synthesis in freshly isolated hepatocytes with greater efficacy than 5-HT. This effect was blocked by olanzapine, an antagonist of 5-HT1/2A receptors. It was mediated by activation of phosphorylase phosphatase, inactivation of glycogen phosphorylase, and activation of glycogen synthase. Unlike insulin action, it was not associated with stimulation of glycolysis and was counteracted by cyclin-dependent kinase (cdk) inhibitors. A role for cdk5 was supported by adaptive changes in the coactivator protein p35 and by elevated glycogen synthesis during overexpression of p35/cdk5. These results support a novel mechanism for serotonin stimulation of hepatic glycogenesis involving cdk5. The opposing effects of serotonin, mediated by distinct 5-HT receptors, could explain why drugs targeting serotonin function can cause either diabetes or hypoglycemia in humans.


Improved cell viability for large-scale biofabrication with photo-crosslinkable hydrogel systems through a dual-photoinitiator approach.
Biofabrication with various hydrogel systems allows the production of tissue or organ constructs in vitro to address various challenges in healthcare and medicine. In particular, photocrosslinkable hydrogels have great advantages such as excellent spatial and temporal selectivity and low processing cost and energy requirements. However, inefficient polymerization kinetics of commercialized photoinitiators upon exposure to UV-A radiation or visible light increase processing time, often compromising cell viability. In this study, we developed a hydrogel crosslinking system which exhibited efficient crosslinking properties and desired mechanical properties with high cell viability, through a dual-photoinitiator approach. Through the co-existence of Irgacure 2959 and VA-086, the overall crosslinking process was completed with a minimal UV dosage during a significantly reduced crosslinking time, producing mechanically robust hydrogel constructs, while most encapsulated cells within the hydrogel constructs remained viable. Moreover, we fabricated a large PEGDA hydrogel construct with a single microchannel as a proof of concept for hydrogels with vasculature to demonstrate the versatility of the system. Our dual-photoinitiator approach allowed the production of this photocrosslinkable hydrogel system with microchannels, significantly improving cell viability and processing efficiency, yet maintaining good mechanical stability. Taken together, we envision the concurrent use of photoinitiators, Irgacure 2959 and VA-086, opening potential avenues for the utilization of various photocrosslinkable hydrogel systems in perfusable large artificial tissue for in vivo and ex vivo applications with improved processing efficiency and cell viability.