More experienced surgeons less likely to fuse: a focus group review of 315 hypothetical EOS cases.
Previous work has identified significant variability in decision making and multiple areas of clinical equipoise in the treatment of early-onset scoliosis (EOS). In an attempt to better understand possible determinants of this variability, we examined the relationship between socioclinical attributes of 11 participating surgeons and decision making regarding the treatment of EOS. Eleven experienced EOS surgeons were surveyed. The first part of the survey consisted of questions regarding surgeon and practice demographics. Next, surgeons were queried regarding their preferred management of 315 hypothetical EOS cases. Cases varied considerably in etiology [idiopathic, and low-tone and high-tone neuromuscular (HTNM)], age, and curve severity and progression. Treatment options were analyzed both individually and grouped as conservative (observation, bracing, or casting) versus surgical (spine-based or rib-based distraction, growth guidance, growth modulation, or definitive fusion). An "outlier" variable was created to determine the extent of a surgeon's deviation from the group in management decisions. A univariate and multivariate regression analysis to identify statistical associations between physician characteristics and their management decisions in the presented hypothetical cases was performed. The cohort's mean years in practice was 20.7±7.36 years. Fifty-six percent of the cohort held Chest Wall and Spine Deformity Study Group (CWSDSG) membership and 56% were members of the Growing Spine Study Group. Multivariate regression demonstrated more years of practice predicted a lower preference for fusion (P<0.05). This effect was greater among HTNM cases (P<0.05). Overall, there was equal interest among groups regarding the choice between rib-based and spine-based distraction methods; however, for the subset of patients with HTNM scoliosis, membership in the CWSDSG (P<0.05) and the percentage of practice spent treating spinal deformity (P<0.05) predicted more rib-based distraction use. EOS surgeons with more experience were less likely to opt for definitive fusion. Use of rib-based distraction methods was common across surgeons in both study groups and within various cohorts of patients. Level V (survey of experts).


Evaluation of genetic variations in the androgen and estrogen metabolic pathways as risk factors for sporadic and familial prostate cancer.
Previous studies suggest that enzymes involved in the androgen metabolic pathway are susceptibility factors for prostate cancer. Estrogen metabolites functioning as genotoxins have also been proposed as risk factors. In this study, we systematically tested the hypothesis that common genetic variations for those enzymes involved in the androgen and estrogen metabolic pathways increase risk for sporadic and familial prostate cancer. From these two pathways, 46 polymorphisms (34 single nucleotide polymorphisms, 10 short tandem repeat polymorphisms, and 2 null alleles) in 25 genes were tested for possible associations. Those genes tested included PRL, LHB, CYP11A1, HSD3B1, HSD3B2, HSD17B2, CYP17, SRD5A2, AKR1C3, UGT2B15, AR, SHBG, and KLK3 from the androgen pathway and CYP19, HSD17B1, CYP1A1, CYP1A2, CYP1B1, COMT, GSTP1, GSTT1, GSTM1, NQO1, ESR1, and ESR2 from the estrogen pathway. A case-control study design was used with two sets of cases: familial cases with a strong prostate cancer family history (n = 438 from 178 families) and sporadic cases with a negative prostate cancer family history (n = 499). The controls (n = 493) were derived from a population-based collection. Our results provide suggestive findings for an association with either familial or sporadic prostate cancer with polymorphisms in four genes: AKR1C3, HSD17B1, NQO1, and GSTT1. Additional suggestive findings for an association with clinical variables (disease stage, grade, and/or node status) were observed for single nucleotide polymorphisms in eight genes: HSD3B2, SRD5A2, SHBG, ESR1, CYP1A1, CYP1B1, GSTT1, and NQO1. However, none of the findings were statistically significant after appropriate corrections for multiple comparisons. Given that the point estimates for the odds ratio for each of these polymorphisms are <2.0, much larger sample sizes will be required for confirmation.


REVIEW: Oxytocin: Crossing the bridge between basic science and pharmacotherapy.
Is oxytocin the hormone of happiness? Probably not. However, this small nine amino acid peptide is involved in a wide variety of physiological and pathological functions such as sexual activity, penile erection, ejaculation, pregnancy, uterus contraction, milk ejection, maternal behavior, osteoporosis, diabetes, cancer, social bonding, and stress, which makes oxytocin and its receptor potential candidates as targets for drug therapy. In this review, we address the issues of drug design and specificity and focus our discussion on recent findings on oxytocin and its heterotrimeric G protein-coupled receptor OTR. In this regard, we will highlight the following topics: (i) the role of oxytocin in behavior and affectivity, (ii) the relationship between oxytocin and stress with emphasis on the hypothalamo-pituitary-adrenal axis, (iii) the involvement of oxytocin in pain regulation and nociception, (iv) the specific action mechanisms of oxytocin on intracellular Ca²(+) in the hypothalamo neurohypophysial system (HNS) cell bodies, (v) newly generated transgenic rats tagged by a visible fluorescent protein to study the physiology of vasopressin and oxytocin, and (vi) the action of the neurohypophysial hormone outside the central nervous system, including the myometrium, heart and peripheral nervous system. As a short nine amino acid peptide, closely related to its partner peptide vasopressin, oxytocin appears to be ideal for the design of agonists and antagonists of its receptor. In addition, not only the hormone itself and its binding to OTR, but also its synthesis, storage and release can be endogenously and exogenously regulated to counteract pathophysiological states. Understanding the fundamental physiopharmacology of the effects of oxytocin is an important and necessary approach for developing a potential pharmacotherapy.


A high-throughput LC-MS/MS assay for quantification of artesunate and its metabolite dihydroartemisinin in human plasma and saliva.
Saliva is an alternative sampling matrix to plasma, offering a noninvasive technique, but requires a highly sensitive bioanalytical method. An API 3000 triple quadrupole mass spectrometer with an electrospray ionization source operated in the positive ion mode was used for the analysis. A high-throughput LC-MS/MS method using SPE for the quantification of artesunate and dihydroartemisinin in plasma and saliva has been optimized and validated according to US FDA guidelines. For both analytes the LLOQ was determined to 5 ng/ml and the calibration range was 5-1000 ng/ml for artesunate and 5-2000 ng/ml for dihydroartemisinin. For the first time, a bioanalytical method for determination of artesunate and dihydroartemisinin in human saliva has been described, showing possible applicability in clinical saliva samples in addition to plasma samples.


Transcobalamins I and II as natural transport proteins of vitamin B12.
There are two conflicting theories of how plasma vitamin B12 (B12) is transported in man: (a) by two distinct transport proteins, transcobalamins I and II (TC I and II), each having a specific role and time of function; and (b) by three active transport proteins, TC I, II, and III, that take up B12 randomly in proportion to the unsaturated amounts of each. To test these theories a man was given 1.12 mug, 229 muCi, of [57Co]B12 mixed with food. Blood samples were taken several times on the 1st day and at lengthening intervals up to day 51. The amount of TC II-B12 was measured in each sample by: gel filtration and by precipitation with (NH4)2SO4. Total serum R-B12 was then separated into TC I and TC III by: (a) a single step anion exchange system and (b) isoelectric focusing (IEF). As the B12 was being absorbed, 92-95% of that in venous blood was carried by TC II. Absolute and percentage transport by TC II declined sharply during the first 24 h; between days 7 and 51 20-33% of the label was on TC II, and the rest was carried by R-type binders. Absolute transport by TC I did not reach a maximum until after day 1 and before day 3. Transport by an alpha2 R-type binder, TC III, could not be demonstrated. TC I was isoelectrically heterogenous, with the components focusing between pH 2.9 and 3.35. It was concluded that (a) TC II is the dominant carrier of B12 immediately after absorption; (b) maximum transport by TC I requires the passage of time after absorption; (c) after the absorbed B12 reaches equilibrium with the total body B12, about one fourth of the plasma B12 is carried by TC II and three fourth by TC I; and (d) TC I and TC II are the only functional transport proteins of plasma B12.


Routine quantitative analysis of brain and cerebrospinal fluid spaces with MR imaging.
A computerized system for processing spin-echo magnetic resonance (MR) imaging data was implemented to estimate whole brain (gray and white matter) and cerebrospinal fluid volumes and to display three-dimensional surface reconstructions of specified tissue classes. The techniques were evaluated by assessing the radiometric variability of MR volume data and by comparing automated and manual procedures for measuring tissue volumes. Results showed (a) the homogeneity of the MR data and (b) that automated techniques were consistently superior to manual techniques. Both techniques, however, were affected by the complexity of the structure, with simpler structures (eg, the intracranial cavity) showing less variability and better spatial correlation of segmentation results between raters. Moreover, the automated techniques were completed for whole brain in a fraction of the time required to complete the equivalent segmentation manually. Additional evaluations included interrater reliability and an evaluation that included longitudinal measurement, in which one subject was imaged sequentially 24 times, with reliability computed from data collected by three raters over 1 year. Results showed good reliability for the automated segmentation procedures.


Serum levels of leptin, zinc and tryptophan with obesity: A case-control study.
The obesity epidemic has turn into a major health threat worldwide and extensively responsible for the increased incidence of many diseases such as diabetes, cardiovascular disease and certain types of cancer. Excessive food intake along with the insufficient physical exercise is the basic impetus for this development. The aim of the present study is to evaluate the serum levels of leptin, zinc and tryptophan (TRP) in obese and non-obese subjects, which play major role in obesity. With the verbal and written consent eighty men were identified from the various areas of Karachi, Pakistan. The socio-demographic data including; age, body mass index (BMI), education and residence, of participants was collected. After providing informed consent, fasting blood samples were taken and serum was collected. The serum concentration of leptin, zinc and TRP were analyzed by ELISA (Enzyme-linked immunosorbent assay), FAAS (Flame atomic absorption spectrophotometer) and HPLC (High performance liquid chromatography) respectively. Results showed that levels of leptin were increased in obese than non-obese subjects significantly. On the other hand levels of zinc and TRP were significantly decreased in obese than non-obese subjects. Furthermore, there was a positive correlation found among leptin, zinc and TRP with obesity. Based on these facts the involvement of leptin, zinc and TRP with obesity will be discussed.


Direct angiotensin type 2 receptor (AT2R) stimulation attenuates T-cell and microglia activation and prevents demyelination in experimental autoimmune encephalomyelitis in mice.
In the present study, we evaluated stimulation of the angiotensin type 2 receptor (AT2R) by the selective non-peptide agonist Compound 21 (C21) as a novel therapeutic concept for the treatment of multiple sclerosis using the model of experimental autoimmune encephalomyelitis (EAE) in mice. C57BL-6 mice were immunized with myelin-oligodendrocyte peptide and treated for 4 weeks with C21 (0.3 mg/kg/day i.p.). Potential effects on myelination, microglia and T-cell composition were estimated by immunostaining and FACS analyses of lumbar spinal cords. The in vivo study was complemented by experiments in aggregating brain cell cultures and microglia in vitro. In the EAE model, treatment with C21 ameliorated microglia activation and decreased the number of total T-cells and CD4+ T-cells in the spinal cord. Fluorescent myelin staining of spinal cords further revealed a significant reduction in EAE-induced demyelinated areas in lumbar spinal cord tissue after AT2R stimulation. C21-treated mice had a significantly better neurological score than vehicle-treated controls. In aggregating brain cell cultures challenged with lipopolysaccharide (LPS) plus interferon-γ (IFNγ), AT2R stimulation prevented demyelination, accelerated re-myelination and reduced the number of microglia. Cytokine synthesis and nitric oxide production by microglia in vitro were significantly reduced after C21 treatment. These results suggest that AT2R stimulation protects the myelin sheaths in autoimmune central nervous system inflammation by inhibiting the T-cell response and microglia activation. Our findings identify the AT2R as a potential new pharmacological target for demyelinating diseases such as multiple sclerosis.


"This is probably the reason why she resorted to that kind of action": A qualitative analysis of juvenile justice workers attributions for girls' offending.
A large body of evidence suggests that within the juvenile justice system, girls fare worse than boys on several measures, including number of arrests, length of stay, and mental health outcomes while in the system. Scholarship suggests a myriad of gendered social factors that precipitate girls' involvement in the juvenile justice system; however, less is known about how stakeholders within the juvenile justice system perceive the girls they work with or interpret their experiences. The current paper examines the attributions that juvenile justice system workers make about the reasons girls offend. In line with previous research, we identify both internal (personality, character traits) and external (situational) explanations for girls' involvement in the juvenile justice system that correspond to gender stereotypes and expectations of girls. Furthermore, we identify structural attributions as a special subset of external attributions that take into account how larger social, economic, and historical factors shape girls' situations and experiences and contribute to their criminal behavior. These structural attributions have implications for practitioners' views of justice and the role of the juvenile justice system in the lives of girls. We conclude with a set of implications for practice and policy.


Sewage sludge, compost and other representative organic wastes as agricultural soil amendments: Benefits versus limiting factors.
Nine different samples of sewage sludges, composts and other representative organic wastes, with potential interest to be used as agricultural soil amendments, were characterized: municipal sewage sludge (SS1 and SS2), agro industrial sludge (AIS), municipal slaughterhouse sludge (MSS), mixed municipal solid waste compost (MMSWC), agricultural wastes compost (AWC), compost produced from agricultural wastes and sewage sludge (AWSSC), pig slurry digestate (PSD) and paper mill wastes (PMW). The characterization was made considering their: (i) physicochemical parameters, (ii) total and bioavailable heavy metals (Cd, Cr, Cu, Ni, Pb, Zn and Hg), (iii) organic contaminants, (iv) pathogenic microorganisms and (v) stability and phytotoxicity indicators. All the sludges, municipal or other, comply with the requirements of the legislation regarding the possibility of their application to agricultural soil (with the exception of SS2, due to its pathogenic microorganisms content), with a content of organic matter and nutrients that make them interesting to be applied to soil. The composts presented, in general, some constraints regarding their application to soil, and their impairment was due to the existence of heavy metal concentrations exceeding the proposed limit of the draft European legislation. As a consequence, with the exception of AWSSC, most compost samples were not able to meet these quality criteria, which are more conservative for compost than for sewage sludge. From the results, the composting of sewage sludge is recommended as a way to turn a less stabilized waste into a material that is no longer classified as a waste and, judging by the results of this work, with lower heavy metal content than the other composted materials, and without sanitation problems.


A characterization of autumn nocturnal migration detected by weather surveillance radars in the northeastern USA.
Billions of birds migrate at night over North America each year. However, few studies have described the phenology of these movements, such as magnitudes, directions, and speeds, for more than one migration season and at regional scales. In this study, we characterize density, direction, and speed of nocturnally migrating birds using data from 13 weather surveillance radars in the autumns of 2010 and 2011 in the northeastern USA. After screening radar data to remove precipitation, we applied a recently developed algorithm for characterizing velocity profiles with previously developed methods to document bird migration. Many hourly radar scans contained windborne "contamination," and these scans also exhibited generally low overall reflectivities. Hourly scans dominated by birds showed nightly and seasonal patterns that differed markedly from those of low reflectivity scans. Bird migration occurred during many nights, but a smaller number of nights with large movements of birds defined regional nocturnal migration. Densities varied by date, time, and location but peaked in the second and third deciles of night during the autumn period when the most birds were migrating. Migration track (the direction to which birds moved) shifted within nights from south-southwesterly to southwesterly during the seasonal migration peaks; this shift was not consistent with a similar shift in wind direction. Migration speeds varied within nights, although not closely with wind speed. Airspeeds increased during the night; groundspeeds were highest between the second and third deciles of night, when the greatest density of birds was migrating. Airspeeds and groundspeeds increased during the fall season, although groundspeeds fluctuated considerably with prevailing winds. Significant positive correlations characterized relationships among bird densities at southern coastal radar stations and northern inland radar stations. The quantitative descriptions of broadscale nocturnal migration patterns presented here will be essential for biological and conservation applications. These descriptions help to define migration phenology in time and space, fill knowledge gaps in avian annual cycles, and are useful for monitoring long-term population trends of migrants. Furthermore, these descriptions will aid in assessing potential risks to migrants, particularly from structures with which birds collide and artificial lighting that disorients migrants.


Added predictive value of night-time blood pressure variability for cardiovascular events and mortality: the Ambulatory Blood Pressure-International Study.
The association of ambulatory blood pressure (BP) variability with mortality and cardiovascular events is controversial. To investigate whether BP variability predicts cardiovascular events and mortality in hypertension, we analyzed 7112 untreated hypertensive participants (3996 men) aged 52±15 years enrolled in 6 prospective studies. Median follow-up was 5.5 years. SD of night-time BP was positively associated with age, body mass index, smoking, diabetes mellitus, and average night-time BP (all P<0.001). In a multivariable Cox model, night-time BP variability was an independent predictor of all-cause mortality (systolic, P<0.001/diastolic, P<0.0001), cardiovascular mortality (P=0.008/<0.0001), and cardiovascular events (P<0.001/<0.0001). In contrast, daytime BP variability was not an independent predictor of outcomes in any model. In fully adjusted models, a night-time systolic BP SD of ≥12.2 mm Hg was associated with a 41% greater risk of cardiovascular events, a 55% greater risk of cardiovascular death, and a 59% increased risk of all-cause mortality compared with an SD of <12.2 mm Hg. The corresponding values for a diastolic BP SD of ≥7.9 mm Hg were 48%, 132%, and 77%. The addition of night-time BP variability to fully adjusted models had a significant impact on risk reclassification and integrated discrimination for all outcomes (relative integrated discrimination improvement for systolic BP variability: 9% cardiovascular events, 14.5% all-cause death, 8.5% cardiovascular death, and for diastolic BP variability: 10% cardiovascular events, 19.1% all-cause death, 23% cardiovascular death, all P<0.01). Thus, addition of BP variability to models of long-term outcomes improved the ability to stratify appropriately patients with hypertension among risk categories defined by standard clinical and laboratory variables.


The histone demethylase JARID1A is associated with susceptibility to ankylosing spondylitis.
Associations with disease identified by genome-wide association studies (GWAS) must be replicated and refined to validate causative variants. In the Wellcome Trust Case Control Consortium (WTCCC) GWAS using 14 500 non-synonymous single nucleotide polymorphisms (nsSNP), rs11062385 (a nsSNP in JARID1A) showed nominal association with ankylosing spondylitis (AS) (P=0.0006, odds ratio (OR)=1.26, 95% confidence interval (95% CI)=1.1-1.4). To replicate and refine the association of JARID1A, rs11062385 was genotyped in 730 further cases and compared with allele frequencies in non-AS disease cohorts typed by WTCCC. We replicated the initial association (P=0.04, OR=1.16, 95% CI=1.01-1.34) and identified a strengthened association with AS in a meta-analysis of this new study combined with the original WTCCC study (P=0.0001, OR=1.21, 95% CI=1.10-1.33). We also genotyped nine further intronic tagging SNPs in JARID1A in 1604 AS cases and 1020 new control samples, but none was associated with AS. JARID1A or a locus in strong linkage disequilibrium with it is a positional candidate for susceptibility to AS.


Decoding the roles of pilotins and accessory proteins in secretin escort services.
Secretins are channels that allow translocation of macromolecules across the outer membranes of Gram-negative bacteria. Virulence, natural competence, and motility are among the functions mediated by these large oligomeric protein assemblies. Filamentous phage also uses secretins to exit their bacterial host without causing cell lysis. However, the secretin is only a part of a larger membrane-spanning complex, and additional proteins are often required for its formation. A class of outer membrane lipoproteins called pilotins has been implicated in secretin assembly and/or localization. Additional accessory proteins may also be involved in secretin stability. Significant progress has recently been made toward deciphering the complex interactions required for functional secretin assembly. To allow for easier comparison between different systems, we have classified the secretins into five different classes based on their requirements for proteins involved in their assembly, localization, and stability. An overview of pilotin and accessory protein structures, functions, and characterized modes of interaction with the secretin is presented.


Backup in gene regulatory networks explains differences between binding and knockout results.
The complementarity of gene expression and protein-DNA interaction data led to several successful models of biological systems. However, recent studies in multiple species raise doubts about the relationship between these two datasets. These studies show that the overwhelming majority of genes bound by a particular transcription factor (TF) are not affected when that factor is knocked out. Here, we show that this surprising result can be partially explained by considering the broader cellular context in which TFs operate. Factors whose functions are not backed up by redundant paralogs show a fourfold increase in the agreement between their bound targets and the expression levels of those targets. In addition, we show that incorporating protein interaction networks provides physical explanations for knockout effects. New double knockout experiments support our conclusions. Our results highlight the robustness provided by redundant TFs and indicate that in the context of diverse cellular systems, binding is still largely functional.


Meta-analysis of magnetic resonance imaging studies of the corpus callosum in bipolar disorder.
The corpus callosum (CC) plays a pivotal role in inter-hemispheric transfer and integration of information and is a relatively understudied structure in bipolar disorder (BD). Magnetic resonance imaging (MRI) studies have reported callosal abnormalities in this condition but findings have been inconsistent. Structural changes affecting the CC may underlie functional abnormalities in BD and could contribute to, or explain the pathophysiology of, the condition. A systematic review was carried out to identify, appraise and summarize MRI studies which compared callosal areas in BD with an unrelated control group. The findings were then synthesized using random effects meta-analysis. Consideration was given to a number of variables to explain heterogeneity. Five case-control studies were identified. Bipolar patients showed reduced callosal areas in comparison with healthy volunteers with no evidence of heterogeneity or publication bias. Findings from this study indicate that callosal areas are reduced in BD and suggest that a failure to integrate information across the hemispheres may contribute to the pathophysiology of the disorder. Further research is necessary to clarify the underlying cellular changes leading to these morphometric differences.


Some like it hot: a differential response to changing temperatures by the malaria vectors Anopheles funestus and An. gambiae s.l.
With the possible implications of global warming, the effect of temperature on the dynamics of malaria vectors in Africa has become a subject of increasing interest. Information from the field is, however, relatively sparse. We describe the effect of ambient temperature over a five-year period on the dynamics of An. funestus and An. gambiae s.l., collected from a single village in southern Mozambique where temperatures varied from a night-time minimum of 6 °C in the cool season to a daytime maximum of 35 °C in the hot season. Mean daily air temperatures varied from 34 °C to 20 °C and soil temperatures varied from 26 °C to 12 °C. Diurnal variation was greatest in the cooler months of the year and were greater in air temperatures than soil temperatures. During the study 301, 705 female An. funestus were collected in 6,043 light-trap collections, 161, 466 in 7,397 exit collections and 16, 995 in 1,315 resting collections. The equivalent numbers for An. gambiae s.l. are 72,475 in light-traps, 33, 868 in exit collections and 5,333 from indoor resting collections. Numbers of mosquito were greatest in the warmer months. Numbers of An. gambiae s.l. went through a one hundredfold change (from a mean of 0.14 mosquitoes a night to 14) whereas numbers of An. funestus merely doubled (from a mean of 20 to 40 a night). The highest environmental correlations and mosquito numbers were between mean air temperature (r2 = 0.52 for An. funestus and 0.77 for An. gambiae s.l.). Numbers of mosquito collected were not related to rainfall with lags of up to four weeks. Numbers of both gravid and unfed An. gambiae complex females in exit collections continued to increase at all temperatures recorded but gravid females of An. funestus decreased at temperatures above 28 °C. Overall the numbers of gravid and unfed An. funestus collected in exit collections were not correlated (p = 0.07). For an unknown reason the number of An. gambiae s.l. fell below monitoring thresholds during the study. Mean air temperature was the most important environmental parameter affecting both vectors in this part of Mozambique. Numbers of An. gambiae s.l. increased at all temperatures recorded whilst An. funestus appeared to be adversely affected by temperatures of 28 °C and above. These differences may influence the distribution of the vectors as the planet warms.


U.S. Attending Anesthesiologist Burnout in the Postpandemic Era.
Anesthesiologists are experiencing unprecedented levels of workplace stress and staffing shortages. This analysis aims to assess how U.S. attending anesthesiologist burnout changed since the onset of the COVID-19 pandemic and target well-being efforts. The authors surveyed the American Society of Anesthesiologists' U.S. attending anesthesiologist members in November 2022. Burnout was assessed using the Maslach Burnout Inventory Human Services Survey with additional questions relating to workplace and demographic factors. Burnout was categorized as high risk for burnout (exhibiting emotional exhaustion and/or depersonalization) or burnout syndrome (demonstrating all three burnout dimensions concurrently). The association of burnout with U.S. attending anesthesiologist retention plans was analyzed, and associated factors were identified. Of 24,680 individuals contacted, 2,698 (10.9%) completed the survey, with 67.7% (1,827 of 2,698) at high risk for burnout and 18.9% (510 of 2,698) with burnout syndrome. Most (78.4%, n = 2,115) respondents have experienced recent staffing shortages, and many (36.0%, n = 970) were likely to leave their job within the next 2 yr. Those likely to leave their job in the next 2 yr had higher prevalence of high risk for burnout (78.5% [760 of 970] vs. 55.7% [651 of 1,169], P < 0.001) and burnout syndrome (24.3% [236 of 970] vs. 13.3% [156 of 1,169], P < 0.001) compared to those unlikely to leave. On multivariable analysis, perceived lack of support at work (odds ratio, 9.2; 95% CI, 7.0 to 12.1), and staffing shortages (odds ratio, 1.96; 95% CI, 1.57 to 2.43) were most strongly associated with high risk for burnout. Perceived lack of support at work (odds ratio, 6.3; 95% CI, 3.81 to 10.4) was the factor most strongly associated with burnout syndrome. Burnout is more prevalent in anesthesiology since early 2020, with workplace factors of perceived support and staffing being the predominant associated variables. Interventions focused on the drivers of burnout are needed to improve well-being among U.S. attending anesthesiologists.


Effects of tillage, crop establishment and diversification on soil organic carbon, aggregation, aggregate associated carbon and productivity in cereal systems of semi-arid Northwest India.
Intensive tillage based management practices are threatening soil quality and systems sustainability in the rice-wheat belt of Northwest India. Furthermore, it is accentuated with puddling of soil, which disrupts soil aggregates. Conservation agriculture (CA) practices involving zero tillage, crop residue management and suitable crop rotation can serve as better alternative to conventional agriculture for maintaining soil quality. Soil organic carbon is an important determinant of soil quality, playing critical role in food production, mitigation and adaptation to climate change as well as performs many ecosystem functions. To understand the turnover of soil carbon in different forms (Total organic carbon-TOC; aggregate associated carbon-AAC; particulate organic carbon- POC), soil aggregation and crop productivity with different management practices, one conventional agriculture based scenario and three CA based crop management scenarios namely conventional rice-wheat system (Sc1), partial CA based rice-wheat-mungbean system (Sc2), full CA-based rice-wheat-mungbean system (Sc3) and maize-wheat-mungbean system (Sc4) were evaluated. TOC was increased by 71%, 68% and 25% after 4 years of the experiment and 75%, 80% and 38% after 6 years of the experiment in Sc4, Sc3 and Sc2, respectively, over Sc1 at 0-15 cm soil depth. After 4 years of the experiment, 38.5% and 5.0% and after 6 years 50.8% and 24.4% improvement in total water stable aggregates at 0-15 and 15-30 cm soil depth, respectively was observed in CA-based scenarios over Sc1. Higher aggregate indices were associated with Sc3 at 0-15 cm soil depth than others. Among the size classes of aggregates, highest aggregate associated C (8.94 g kg-1) was retained in the 1-0.5 mm size class under CA-based scenarios. After 6 years, higher POC was associated with Sc4 (116%). CA-based rice/maize system (Sc3 and Sc4) showed higher productivity than Sc1. Therefore, CA could be a potential management practice in rice-wheat cropping system of Northwest India to improve the soil carbon pools through maintaining soil aggregation and productivity.


Gold nanoparticles supported by imidazolium-based porous organic polymers for nitroarene reduction.
Two imidazolium-based porous organic polymers (Im-POP-1 and Im-POP-2) were synthesized through the Yamamoto reaction of different molar ratios of 1,3-bis(4-bromophenyl)imidazolium bromide and tetrakis(4-bromophenyl)methane, subsequent anion exchange with HAuCl4 and reduction with NaBH4 produced well-dispersed gold nanoparticles (NPs), Au@Im-POP-1 and Au@Im-POP-2. These NPs possess a small size and narrow size distribution, their application in the reduction of nitroarenes was evaluated, and the reduction process was tracked by 13C NMR experiments for reaction mechanism studies. In comparison with gold NPs supported by a non-ionic analogue from the Yamamoto reaction of tetrakis(4-bromophenyl)methane (Au@POP-TPM), Au@Im-POP-2 exhibits high catalytic activity, good selectivity of aniline and superior recyclability in nitrobenzene reduction. The gold NPs in Au@Im-POP-2 increase slightly after four consecutive reaction runs, while an apparent agglomeration of NPs in Au@POP-TPM was observed after the second run. This research provides a new type of support for the stabilization of gold NPs and for the improvement of catalytic performances.


RAD51 paralogs: roles in DNA damage signalling, recombinational repair and tumorigenesis.
Chromosomal double-strand breaks (DSBs) have the potential to permanently arrest cell cycle progression and endanger cell survival. They must therefore be efficiently repaired to preserve genome integrity and functionality. Homologous recombination (HR) provides an important error-free mechanism for DSB repair in mammalian cells. In addition to RAD51, the central recombinase activity in mammalian cells, a family of proteins known as the RAD51 paralogs and consisting of five proteins (RAD51B, RAD51C, RAD51D, XRCC2 and XRCC3), play an essential role in the DNA repair reactions through HR. The RAD51 paralogs act to transduce the DNA damage signal to effector kinases and to promote break repair. However, their precise cellular functions are not fully elucidated. Here we discuss recent advances in our understanding of how these factors mediate checkpoint responses and act in the HR repair process. In addition, we highlight potential functional similarities with the BRCA2 tumour suppressor, through the recently reported links between RAD51 paralog deficiencies and tumorigenesis triggered by genome instability.


Specific flavonoids promote intercellular root colonization of Arabidopsis thaliana by Azorhizobium caulinodans ORS571.
The ability of Azorhizobium caulinodans ORS571 and other diazotrophic bacteria to internally colonize roots of Arabidopsis thaliana has been studied. Strains tagged with lacZ or gusA reporter genes were used, and the principal colonization sites were found to be the points of emergence of lateral roots, lateral root cracks (LRCs). High frequencies of colonization were found; 63 to 100% of plants were colonized by ORS571, and 100% of plants were colonized by Herbaspirillum seropedicae. After LRCs were colonized, bacteria moved into intercellular spaces between the cortical and endodermal cell layers. Specific flavonoids, naringenin and daidzein, at 5 x 10(-5) M, significantly promoted colonization by ORS571. By using a nodC and a nodD mutant of ORS571, it was shown that neither Nod factors nor NodD are involved in colonization or flavonoid stimulation of colonization. Flavonoids did not appear to be stimulating LRC colonization by their activity as nutritional factors. LRC and intercellular colonization by H. seropedicae was stimulated by naringenin and daidzein at the same concentration that stimulated colonization by ORS571.


Molecular and Cellular Mechanisms of Melatonin in Osteosarcoma.
Osteosarcoma, the most common primary bone malignancy, occurs most frequently in adolescents with a peak of incidence at 11-15 years. Melatonin, an indole amine hormone, shows a wide range of anticancer activities. The decrease in melatonin levels simultaneously concurs with the increase in bone growth and the peak age distribution of osteosarcoma during puberty, so melatonin has been utilized as an adjunct to chemotherapy to improve the quality of life and clinical outcomes. While a large amount of research has been conducted to understand the complex pleiotropic functions and the molecular and cellular actions elicited by melatonin in various types of cancers, a few review reports have focused on osteosarcoma. Herein, we summarized the anti-osteosarcoma effects of melatonin and its underlying molecular mechanisms to illustrate the known significance of melatonin in osteosarcoma and to address cellular signaling pathways of melatonin in vitro and in animal models. Even in the same kind of osteosarcoma, melatonin has been sparingly investigated to counteract tumor growth, apoptosis, and metastasis through different mechanisms, depending on different cell lines. We highlighted the underlying mechanism of anti-osteosarcoma properties evoked by melatonin, including antioxidant activity, anti-proliferation, induction of apoptosis, and the inhibition of invasion and metastasis. Moreover, we discussed the drug synergy effects of the role of melatonin involved and the method to fortify the anti-cancer effects on osteosarcoma. As a potential therapeutic agent, melatonin is safe for children and adolescents and is a promising candidate for an adjuvant by reinforcing the therapeutic effects and abolishing the unwanted consequences of chemotherapies.


Accounting for the influence of inflammation on retinol-binding protein in a population survey of Liberian preschool-age children.
Vitamin A deficiency (VAD) is an important contributor to child morbidity and mortality. The prevalence of VAD, measured by retinol-binding protein (RBP) or retinol, is overestimated in populations with a high prevalence of inflammation. We aimed to quantify and adjust for the effect of inflammation on VAD prevalence in a nationally representative survey of Liberian children 6 to 35 months of age. We compared five approaches to adjust RBP for inflammation and estimate VAD prevalence (defined as RBP < 0.7 µmol/L): (1) ignoring inflammation; (2) excluding individuals with inflammation (C-reactive protein (CRP) >5 mg/L or alpha1-acid glycoprotein (AGP) >1 g/)L; (3) multiplying each individual's RBP by an internal correction factor; (4) by an external correction factor; and (5) using regression (corrected RBP = exp(InRBP - β1 (lnCRPobs -lnCRPref ) - β2 (lnAGPobs -lnAGPref )). Corrected RBP was based on a regression model where reference lnCRP and lnAGP were set to the maximum of the lowest decile. The unadjusted prevalence of VAD was 24.7%. Children with elevated CRP and/or AGP had significantly lower RBP concentrations than their apparently healthy peers (geometric mean RBP 0.79 µmol/L (95% CI: 0.76, 0.82) vs. 0.95 µmol/L (95% CI: 0.92, 0.97), P < 0.001). Using approaches 2-5 resulted in a prevalence of VAD of 11.6%, 14.3%, 13.5% and 7.3%, respectively. Depending on the approach, the VAD prevalence is reduced 10-17 percentage points when inflammation is taken into account. Further quantification of the influence of inflammation on biomarkers of vitamin A status from other national surveys is needed to compare and recommend the preferred adjustment approach across populations.


One Standardized Differentiation Procedure Robustly Generates Homogenous Hepatocyte Cultures Displaying Metabolic Diversity from a Large Panel of Human Pluripotent Stem Cells.
Human hepatocytes display substantial functional inter-individual variation regarding drug metabolizing functions. In order to investigate if this diversity is mirrored in hepatocytes derived from different human pluripotent stem cell (hPSC) lines, we evaluated 25 hPSC lines originating from 24 different donors for hepatic differentiation and functionality. Homogenous hepatocyte cultures could be derived from all hPSC lines using one standardized differentiation procedure. To the best of our knowledge this is the first report of a standardized hepatic differentiation procedure that is generally applicable across a large panel of hPSC lines without any adaptations to individual lines. Importantly, with regard to functional aspects, such as Cytochrome P450 activities, we observed that hepatocytes derived from different hPSC lines displayed inter-individual variation characteristic for primary hepatocytes obtained from different donors, while these activities were highly reproducible between repeated experiments using the same line. Taken together, these data demonstrate the emerging possibility to compile panels of hPSC-derived hepatocytes of particular phenotypes/genotypes relevant for drug metabolism and toxicity studies. Moreover, these findings are of significance for applications within the regenerative medicine field, since our stringent differentiation procedure allows the derivation of homogenous hepatocyte cultures from multiple donors which is a prerequisite for the realization of future personalized stem cell based therapies.


New perspectives in the renin-angiotensin-aldosterone system (RAAS) II: albumin suppresses angiotensin converting enzyme (ACE) activity in human.
About 8% of the adult population is taking angiotensin-converting enzyme (ACE) inhibitors to treat cardiovascular disease including hypertension, myocardial infarction and heart failure. These drugs decrease mortality by up to one-fifth in these patients. We and others have reported previously that endogenous inhibitory substances suppress serum ACE activity, in vivo, similarly to the ACE inhibitor drugs. Here we have made an effort to identify this endogenous ACE inhibitor substance. ACE was crosslinked with interacting proteins in human sera. The crosslinked products were immunoprecipitated and subjected to Western blot. One of the crosslinked products was recognized by both anti-ACE and anti-HSA (human serum albumin) antibodies. Direct ACE-HSA interaction was confirmed by binding assays using purified ACE and HSA. HSA inhibited human purified (circulating) and human recombinant ACE with potencies (IC50) of 5.7 ± 0.7 and 9.5 ± 1.1 mg/mL, respectively. Effects of HSA on the tissue bound native ACE were tested on human saphenous vein samples. Angiotensin I evoked vasoconstriction was inhibited by HSA in this vascular tissue (maximal force with HSA: 6.14 ± 1.34 mN, without HSA: 13.54 ± 2.63 mN), while HSA was without effects on angiotensin II mediated constrictions (maximal force with HSA: 18.73 ± 2.17 mN, without HSA: 19.22 ± 3.50 mN). The main finding of this study is that HSA was identified as a potent physiological inhibitor of the ACE. The enzymatic activity of ACE appears to be almost completely suppressed by HSA when it is present in its physiological concentration. These data suggest that angiotensin I conversion is limited by low physiological ACE activities, in vivo.


Fervidobacterium riparium sp. nov., a thermophilic anaerobic cellulolytic bacterium isolated from a hot spring.
A novel obligately anaerobic, extremely thermophilic, organotrophic bacterium, strain 1445t(T), was isolated from a hot spring on Kunashir Island (Kuril Islands, Russia). Cells were motile rods (0.4-0.5 × 1.0-3.0 µm). The temperature range for growth at pH 7.8 was 46-80 °C, with optimum growth at 65 °C. The pH range for growth at 65 °C was pH 5.7-9.0, with optimum growth at pH 7.8. Growth was not observed at or below 40 °C, at or above 84 °C, at or below pH 5.4 or at or above pH 9.5. The isolate degraded a wide range of substrates including starch, cellulose and cellulose derivatives. Elemental sulfur stimulated growth, but sodium sulfate, sulfite and thiosulfate did not. DNA G+C content was 31 mol%. Phylogenetic analysis of 16S rRNA gene sequences showed that strain 1445t(T) belonged to the genus Fervidobacterium. 16S rRNA gene sequence similarities with strains of other species of the genus Fervidobacterium were 94.9-98.3 %; the type strain of Fervidobacterium gondwanense was the closest relative of strain 1445t(T). DNA-DNA hybridization of strain 1445t(T) and F. gondwanense AB39(T) revealed a relatedness value of 20 %. Based on phylogenetic data and physiological properties of the isolate, a novel species, designated Fervidobacterium riparium sp. nov., is proposed with strain 1445t(T) ( = DSM 21630(T) = VKM B-2549(T)) as the type strain.


Associations Between Movement Behaviors and Emotional Changes in Toddlers and Preschoolers During Early Stages of the COVID-19 Pandemic in Chile.
Background: There is limited evidence about emotional and behavioral responses in toddlers and preschoolers during the novel coronavirus (COVID-19) pandemic, particularly in Latin America. Objective: To assess associations between changes in movement behaviors (physical activity, screen time and sleeping) and emotional changes in toddlers and preschoolers during early stages of the pandemic in Chile. Methods: A cross-sectional study conducted from March 30th to April 27th, 2020. Main caregivers of 1- to 5-year-old children living in Chile answered an online survey that included questions about sociodemographic characteristics, changes in the child's emotions and behaviors, movement behaviors and caregivers' stress during the pandemic. Multiple linear regressions were used to assess the association between different factors and emotional changes in toddlers and preschoolers. Results: In total, 1727 caregivers provided complete data on emotional changes for children aged 2.9 ± 1.36 years old, 47.9% girls. A large proportion of toddlers and preschoolers in Chile experienced emotional and behavioral changes. Most caregivers reported that children "were more affectionate" (78.9%), "more restless" (65.1%), and 'more frustrated' (54.1%) compared with pre-pandemic times. Apart from changes in movement behaviors, factors such as child age, caregivers' age and stress, and residential area (urban/rural) were consistently associated with changes in emotions and behaviors. Conclusion: The pandemic substantially affected the emotions and behaviors of toddlers and preschoolers in Chile. The findings suggest that supportive actions for caregivers may have a positive impact not only on adults but also on children. Mental health promotion programs should consider multilevel approaches in which the promotion of movement behaviors and support for caregivers should be essential pieces for future responses.


Carbohydrate Mouth Rinsing: Improved Neuromuscular Performance During Isokinetic Fatiguing Exercise.
To determine whether repeated carbohydrate (CHO) mouth rinsing would improve neuromuscular performance during high-intensity fatiguing contractions. Eighteen young men (age 26.1 ± 5.0 y, BMI 22.9 ± 1.9) performed 3 maximal voluntary isometric contractions (MVICPRE). Immediately after, they completed 10-second mouth rinse with 6.4% maltodextrin solution (MAL), 7.1% glucose solution (GLU), water (W), artificially sweetened solution (PLA), or a control trial with no rinse (CON) in a crossover protocol. Subjects performed 5 sets of 30 isokinetic fatiguing contractions at 180°/s, and an MVICPOST with their elbow flexors was performed after each mouth rinse. Mechanical and electromyographic (EMG) signals were recorded from the biceps brachii and parameters of interest analyzed. When rinsing the mouth with a solution containing CHO, independently of the sweetness, isokinetic performance was enhanced as shown by the greater total work achieved in comparison with CON. The decay of torque and mean fiber-conduction velocity (MFCV) recorded at the end of the fatiguing task was lower when rinsing the mouth with GLU than with CON. The torque recorded during the MVICPOST was greater with CHO with respect to CON, and this was associated to a lower decay of MFCV. CHO mouth rinse counteracts fatigue-induced decline in neuromuscular performance, supporting the notion that CHO rinse may activate positive afferent signals able to modify motor output. Repeated mouth rinsing with sweet and nonsweet CHO-containing solutions can improve neuromuscular performance during an isokinetic intermittent fatiguing task.


Cord blood collection before and after placental delivery: levels of nucleated cells, haematopoietic progenitor cells, leukocyte subpopulations and macroscopic clots.
The number of nucleated cells infused into the recipient of a cord blood (CB) transplant has emerged as the most important factor affecting the probability and speed of engraftment. At present, there is no international consensus on the procedure of CB collection in the maternity ward. In order to maximise the yield of viable cells in a CB unit, we aimed to investigate the efficiency of CB collection, with respect to the time of delivery of the placenta. We analysed stem and progenitor cells in terms of CD34+ cell content and colony-forming activities, lymphocyte subpopulations and the presence of macroscopic clots in 93 paired CB samples, collected before and after the delivery of the placenta. Our results demonstrated that the median concentrations of nucleated cells and total colony-forming unit (CFU) were significantly lower in CB collected after placenta delivery by 9.5% (P < 0.001) and 11.6% (P = 0.015), respectively, when compared to their counterparts collected before placental delivery. A reduction of granulocytes (P < 0.001), monocytes (P < 0.001) and CD19+ B lymphocytes (P = 0.031) was observed, with no significant change in the proportion of T cell subsets (CD4+, CD8+ cells) or activated T cells (CD25+, CD45RO+ cells) in samples collected after placenta delivery. The incidence of macroscopic clots was also higher in these samples (31% vs 1%, P < 0.001). The reduction of stem and progenitor cells correlated significantly with that of major cell populations, indicating a general cell loss, possibly due to clotting activities developed with time. Our study has documented strong evidence for recommending the collection of CB before the delivery of the placenta.


Defining the epigenetic actions of growth hormone: acute chromatin changes accompany GH-activated gene transcription.
Many of the long-term physiological effects of GH require hormone-mediated changes in gene expression. The transcription factor signal transducer and activator of transcription 5b (Stat5b) plays a critical role in the actions of GH on growth and metabolism by regulating a large number of GH-dependent genes by incompletely understood mechanisms. Here we have assessed the impact of GH-initiated and Stat5b-mediated signaling on the chromatin landscape of hormone-regulated genes in the liver of pituitary-deficient young adult male rats. In the absence of GH there was minimal ongoing transcription at the Socs2, Cish, Igfals, and Spi 2.1 promoters, minimal occupancy of Stat5b at proximal promoter sites, and relatively closed chromatin, as evidenced by low levels of core histone acetylation. In contrast, transcriptionally silent Igf1 promoter 1 appeared poised to be activated, based on binding of coactivators p300 and Med1/Trap220, high levels of histone acetylation, and the presence of RNA polymerase II. GH treatment led to a 8- to 20-fold rise in transcriptional activity of all five genes within 30-60 min and was accompanied by binding of Stat5b to the proximal Socs2, Cish, Igfals, and Spi 2.1 promoters and to seven distal Igf1 Stat5b elements, by enhanced histone acetylation at all five promoters, by recruitment of RNA polymerase II to the Socs2, Cish, Igfals, and Spi 2.1 promoters, and by loss of the transcriptional repressor Bcl6 from Socs2, Cish, and Igfals Stat5b sites, but not from two Igf1 Stat5b domains. We conclude that GH actions induce rapid and dramatic changes in hepatic chromatin at target promoters and propose that the chromatin signature of Igf1 differs from other GH-and Stat5b-dependent genes.


Complexities in Identifying and Defining Mathematics Learning Disability in the Primary School-Age Years.
This paper is a descriptive report of findings from a prospective longitudinal study of math disability (MD). The study was designed to address the incidence of MD during primary school, the utility of different MD definitions, and evidence of MD subtypes. The results illustrate the dynamic properties of psychometrically derived definitions of MD. Different groups of children meet criteria for MD depending on which measure(s) are used for identification. Over time, a given individual may not continue to meet MD criteria, even when using the same assessments. Thus, the findings lead to cautions regarding the single-tool/ one-time assessment for a clinical diagnosis of MD. Twenty-two of 209 participants demonstrated "persistent MD" (MD-p), or MD for more than one school grade. Reading disability was relatively more frequent in this MD-p subgroup than in the remaining participants (25 percent vs. 7 percent). Reading-related skills were correlated with math achievement, as were select visual spatial skills. There was minimal overlap between groups who met either a "poor achievement" criteria or an "IQ-achievement discrepancy," and the latter was far less stable a measure over time than the former. The results highlight the complexities of defining MD and illustrate the need for more research in this area.


Dietary cost associated with adherence to the Mediterranean diet, and its variation by socio-economic factors in the UK Fenland Study.
High cost of healthy foods could be a barrier to healthy eating. We aimed to examine the association between dietary cost and adherence to the Mediterranean diet in a non-Mediterranean country. We evaluated cross-sectional data from 12 417 adults in the UK Fenland Study. Responses to 130-item FFQ were used to calculate a Mediterranean diet score (MDS). Dietary cost was estimated by matching food consumption data with retail prices of five major supermarkets. Using multivariable-adjusted linear regression, we examined the association of MDS and individual foods with dietary cost in absolute and relative scales. Subsequently, we assessed how much the association was explained by education, income, marital status and occupation, by conducting mediation analysis and testing interaction by these variables. High compared with low MDS (top to bottom third) was associated with marginally higher cost by 5·4 % (95 % CI 4·4, 6·4) or £0·20/d (95 % CI 0·16, 0·25). Participants with high adherence had higher cost associated with the healthier components (e.g. vegetables, fruits and fish), and lower cost associated with the unhealthy components (e.g. red meat, processed meat and sweets) (P for trend<0·001 each). In total, 20·7 % (95 % CI 14·3, 27·0) of the MDS-cost association was explained by the selected socio-economic factors, and the MDS-cost association was of greater magnitude in lower socio-economic groups (P interaction<0·005). Overall, greater adherence to the Mediterranean diet was associated with marginally higher dietary cost, partly modified and explained by socio-economic status, but the potential economic barriers of high adherence might be offset by cost saving from reducing unhealthy food consumption.


A modular PIP2 binding site as a determinant of capsaicin receptor sensitivity.
The capsaicin receptor (TRPV1), a heat-activated ion channel of the pain pathway, is sensitized by phosphatidylinositol-4,5-bisphosphate (PIP2) hydrolysis after phospholipase C activation. We identify a site within the C-terminal domain of TRPV1 that is required for PIP2-mediated inhibition of channel gating. Mutations that weaken PIP2-TRPV1 interaction reduce thresholds for chemical or thermal stimuli, whereas TRPV1 channels in which this region is replaced with a lipid-binding domain from PIP2-activated potassium channels remain inhibited by PIP2. The PIP2-interaction domain therefore serves as a critical determinant of thermal threshold and dynamic sensitivity range, tuning TRPV1, and thus the sensory neuron, to appropriately detect heat under normal or pathophysiological conditions.


N-butyldeoxynojirimycin is a novel inhibitor of glycolipid biosynthesis.
The imino sugar deoxynojirimycin and its alkylated derivatives are inhibitors of the N-linked oligosaccharide processing enzymes alpha-glucosidase I and II. These compounds are glucose analogues and have the potential to inhibit both glucosidases and glucosyltransferases. However, to date there has been no report of deoxynojirimycin or similar analogues inhibiting a mammalian glucosyltransferase. We have investigated the effects of deoxynojirimycin and its alkylated derivatives on the biosynthesis of glycolipids in HL-60 cells. We have found that the N-butyl and N-hexyl derivatives of deoxynojirimycin, but not deoxynojirimycin itself, are novel inhibitors of the glucosyltransferase-catalyzed biosynthesis of glucosylceramide. This results in the inhibition of biosynthesis of all glucosylceramide-based glycosphingolipids. We have investigated the ability of one of these compounds, N-butyldeoxynojirimycin, to offset glucosylceramide accumulation in an in vitro Gaucher's disease model. This compound prevents lysosomal glycolipid storage and offers a novel therapeutic approach for the management of this and other glycolipid storage disorders.


Palladium-catalyzed enolate arylation as a key C-C bond-forming reaction for the synthesis of isoquinolines.
The palladium-catalyzed coupling of an enolate with an ortho-functionalized aryl halide (an α-arylation) furnishes a protected 1,5-dicarbonyl moiety that can be cyclized to an isoquinoline with a source of ammonia. This fully regioselective synthetic route tolerates a wide range of substituents, including those that give rise to the traditionally difficult to access electron-deficient isoquinoline skeletons. These two synthetic operations can be combined to give a three-component, one-pot isoquinoline synthesis. Alternatively, cyclization of the intermediates with hydroxylamine hydrochloride engenders direct access to isoquinoline N-oxides; and cyclization with methylamine, gives isoquinolinium salts. Significant diversity is available in the substituents at the C4 position in four-component, one-pot couplings, by either trapping the in situ intermediate after α-arylation with carbon or heteroatom-based electrophiles, or by performing an α,α-heterodiarylation to install aryl groups at this position. The α-arylation of nitrile and ester enolates gives access to 3-amino and 3-hydroxyisoquinolines and the α-arylation of tert-butyl cyanoacetate followed by electrophile trapping, decarboxylation and cyclization, C4-functionalized 3-aminoisoquinolines. An oxime directing group can be used to direct a C-H functionalization/bromination, which allows monofunctionalized rather than difunctionalized aryl precursors to be brought through this synthetic route.


Distinct patterns of thought mediate the link between brain functional connectomes and well-being.
Ongoing thought patterns constitute important aspects of both healthy and abnormal human cognition. However, the neural mechanisms behind these daily experiences and their contribution to well-being remain a matter of debate. Here, using resting-state fMRI and retrospective thought sampling in a large neurotypical cohort (n = 211), we identified two distinct patterns of thought, broadly describing the participants' current concerns and future plans, that significantly explained variability in the individual functional connectomes. Consistent with the view that ongoing thoughts are an emergent property of multiple neural systems, network-based analysis highlighted the central importance of both unimodal and transmodal cortices in the generation of these experiences. Importantly, while state-dependent current concerns predicted better psychological health, mediating the effect of functional connectomes, trait-level future plans were related to better social health, yet with no mediatory influence. Collectively, we show that ongoing thoughts can influence the link between brain physiology and well-being.


Learning Instance-Specific Predictive Models.
This paper introduces a Bayesian algorithm for constructing predictive models from data that are optimized to predict a target variable well for a particular instance. This algorithm learns Markov blanket models, carries out Bayesian model averaging over a set of models to predict a target variable of the instance at hand, and employs an instance-specific heuristic to locate a set of suitable models to average over. We call this method the instance-specific Markov blanket (ISMB) algorithm. The ISMB algorithm was evaluated on 21 UCI data sets using five different performance measures and its performance was compared to that of several commonly used predictive algorithms, including nave Bayes, C4.5 decision tree, logistic regression, neural networks, k-Nearest Neighbor, Lazy Bayesian Rules, and AdaBoost. Over all the data sets, the ISMB algorithm performed better on average on all performance measures against all the comparison algorithms.


Dementia is associated with iron-deficiency anemia in females: a population-based study.
Cognitive derangement and neurological symptoms are observed in patients with anemia. Although it is still controversial, a few studies suggested that anemia may increase the risk of dementia. This study aimed to explore the association between iron-deficiency anemia (IDA) and dementia in a population-based case-control study. We retrieved our study sample from the Taiwan Longitudinal Health Insurance Database 2000. We extracted 8300 subjects with a diagnosis of dementia and 8300 age- and gender-matched controls. The results showed that there was a significant difference in the prevalence of prior IDA between cases and controls (6.0% vs. 3.8%, p<0.001). The conditional logistic regression analysis suggested that the odds ratio (OR) of prior IDA for cases was 1.36 (95% CI: 1.07-1.74) compared to controls after adjusting for subjects' monthly income, geographic location, urbanization level, hyperlipidemia, diabetes, hypertension, and alcohol abuse. Among female subjects, the adjusted OR of prior IDA for cases was as high as 2.00 (95% CI: 1.42-2.80) compared to controls. However, in men-no increased odds of prior IDA were observed, compared to controls. We concluded that women with dementia had a higher prevalence of prior IDA, compared to controls.


Combined Raman and AFM detection of changes in HeLa cervical cancer cells induced by CeO2 nanoparticles - molecular and morphological perspectives.
The design of nanoparticles for application in medical diagnostics and therapy requires a thorough understanding of various aspects of nanoparticle-cell interactions. In this work, two unconventional methods for the study of nanoparticle effects on cells, Raman spectroscopy and atomic force microscopy (AFM), were employed to track the molecular and morphological changes that are caused by the interaction between cervical carcinoma-derived HeLa cells and two types of cerium dioxide (CeO2) nanoparticles, ones with dextran coating and the others with no coating. Multivariate statistical analyses of Raman spectra, such as principal component analysis and partial least squares regression, were applied in order to extract the variations in the vibrational features of cell biomolecules and through them, the changes in biomolecular content and conformation. Both types of nanoparticles induced changes in DNA, lipid and protein contents of the cell and variations of the protein secondary structure, whereas dextran-coated CeO2 affected the cell-growth rate to a higher extent. Atomic force microscopy showed changes in cell roughness, cell height and nanoparticle effects on surface molecular layers. The method differentiated between the impact of dextran-coated and uncoated CeO2 nanoparticles with higher precision than performed viability tests. Due to the holistic approach provided by vibrational information on the overall cell content, accompanied by morphological modifications observed by high-resolution microscopy, this methodology offers a wider picture of nanoparticle-induced cell changes, in a label-free single-cell manner.


Differences in Cancer Death Risk Long After ACS Among Selected Urban and Rural Areas in North Italy: The ABC-7a Study on Heart Disease.
An increased risk of cancer death has been demonstrated for patients diagnosed with acute coronary syndrome (ACS). We are investigating possible geographic risk disparities. This prospective study included 541 ACS patients who were admitted to hospitals and discharged alive in three provinces of Italy's Veneto region. The patients were classified as residing in urban or rural areas in each province. With 3 exceptions, all patients completed the 22-year follow-up or were followed until death. Urban (46%) and rural (54%) residents shared most of their baseline demographic and clinical characteristics. Pre-existing malignancy was noted in 15 patients, whereas 106 patients developed cancer during the follow-up period, which represented 6232 person-years. No difference in the cancer death risk was found between the urban and rural areas or between southern and northern provinces (hazard ratio [HR] 1.1; 95% confidence interval [CI] 0.7-1.7; p = 0.59 and HR 1.1; 95% CI 0.9-1.4; p = 0.29, respectively) according to the unadjusted Cox regression analysis. Geographic areas, however, showed a strong positive interaction, with risk increasing from the urban to rural areas from southern to northern provinces (HR 1.9; 95% CI 1.1-3.0; p = 0.01). The fully adjusted Cox regression and Fine-Gray competing risk regression models provided similar results. Interestingly, these results persisted, and even strengthened, after exclusion of the 22 patients who developed malignancy and survived to the end of follow-up. We did not observe an urban/rural difference in non-neoplastic death risk or a significant interaction between the geographic areas. Our analysis reveals that the cancer death risk among unselected ACS patients in Italy's Veneto region significantly differs by geography. The northern rural area has the highest risk. These results highlight the importance of implementing a preventive policy based on area-specific knowledge.


Effects of transportation during the hot season, breed and electrical stimulation on histochemical and meat quality characteristics of goat longissimus muscle.
The effects of transportation and electrical stimulation (90 V) on physiological, histochemical and meat quality characteristics of two breeds of Omani goats were assessed. Twenty 1-year-old male goats from each breed (Batina and Dhofari) were divided into two groups: 3 h transported during the hot season (42 degrees C day time temperature) and non-transported. Animals were blood-sampled before loading and prior to slaughter. Electrical stimulation was applied 20 min postmortem to 50% randomly selected carcasses of both breeds. Temperature and pH decline of the Longissimus was monitored. Ultimate pH, shear force, sarcomere length, myofibrillar fragmentation index, expressed juice, cooking loss and colour were measured from samples of Longissimus dorsi muscles. Electrical stimulation and transportation had a significant effect on most biochemical and meat quality characteristics of Longissimus dorsi. The transported goats had higher plasma cortisol (P < 0.01), adrenaline, nor-adrenaline and dopamine concentrations (P < 0.05) than non-transported goats. Electrical stimulation resulted in a significantly (P < 0.05) more rapid muscle pH fall during the first 12 h after slaughter. Muscles from electrically-stimulated carcasses had significantly (P < 0.05) longer sarcomeres, lower shear force value, a lighter colour (higher L* value), higher expressed juice and myofibrillar fragmentation index than those from non-stimulated ones. Meat from transported goats had significantly higher pH, expressed juice and shear force, but contained significantly lower sarcomere length and L* values than non-transported goats. The proportion of the myosin ATPase staining did not change as a function of stimulation, transportation or breed. These results indicated that subjecting goats to transportation for 3 h under high ambient temperatures can generate major physiological and muscle metabolism responses. Electrical stimulation improved quality characteristics of meat from both groups. This indicates that electrical stimulation may reduce detrimental effects of transportation on meat quality of Omani goats.


The lysosomal signaling anchor p18/LAMTOR1 controls epidermal development by regulating lysosome-mediated catabolic processes.
The lysosomal adaptor protein p18 is an essential anchor of a scaffolding complex for the mTORC1 and MAPK pathways, which play crucial roles in controlling cell growth and energy homeostasis. To elucidate the in vivo function of the p18-mediated pathway, we conditionally ablated p18 in the mouse epidermis. Mutant mice were born with severe defects in formation of the stratum corneum and died within 12 h after birth due to dehydration caused by loss of skin barrier function. Mutant epidermal cells can grow and differentiate into granular cells, but exhibit functional defects in corneocyte maturation. Electron microscopy identified abnormal immature cells, overlying the mutant granular cells, which accumulated autophagosomes, glycogen granules and dead nuclei. Cell culture analysis showed that loss of p18 attenuated lysosome function, resulting in accumulation of immature lysosomes and autophagosomes. Analyses of lysosome behavior revealed that p18 is required for functional interaction between lysosomes and target organelles including autophagosomes. These findings suggest that p18-mediated pathways control lysosome-mediated catabolic processes, which are crucial for the development of mouse epidermis.


Interactions between Callous Unemotional Behaviors and Executive Function in Early Childhood Predict later Aggression and Lower Peer-liking in Late-childhood.
Callous unemotional (CU) behaviors are linked to aggression, behavior problems, and difficulties in peer relationships in children and adolescents. However, few studies have examined whether early childhood CU behaviors predict aggression or peer-rejection during late-childhood or potential moderation of this relationship by executive function. The current study examined whether the interaction of CU behaviors and executive function in early childhood predicted different forms of aggression in late-childhood, including proactive, reactive, and relational aggression, as well as how much children were liked by their peers. Data from cross-informant reports and multiple observational tasks were collected from a high-risk sample (N = 240; female = 118) at ages 3 and 10 years old. Parent reports of CU behaviors at age 3 predicted teacher reports of reactive, proactive, and relational aggression, as well as lower peer-liking at age 10. Moderation analysis showed that specifically at high levels of CU behaviors and low levels of observed executive function, children were reported by teachers as showing greater reactive and proactive aggression, and were less-liked by peers. Findings demonstrate that early childhood CU behaviors and executive function have unique main and interactive effects on both later aggression and lower peer-liking even when taking into account stability in behavior problems over time. By elucidating how CU behaviors and deficits in executive function potentiate each other during early childhood, we can better characterize the emergence of severe and persistent behavior and interpersonal difficulties across development.


Brain Tumor Epidemiology - A Hub within Multidisciplinary Neuro-oncology. Report on the 15th Brain Tumor Epidemiology Consortium (BTEC) Annual Meeting, Vienna, 2014.
The Brain Tumor Epidemiology Consortium (BTEC) is an open scientific forum, which fosters the development of multi-center, international and inter-disciplinary collaborations. BTEC aims to develop a better understanding of the etiology, outcomes, and prevention of brain tumors (http://epi.grants.cancer.gov/btec/). The 15th annual Brain Tumor Epidemiology Consortium Meeting, hosted by the Austrian Societies of Neuropathology and Neuro-oncology, was held on September 9 - 11, 2014 in Vienna, Austria. The meeting focused on the central role of brain tumor epidemiology within multidisciplinary neuro-oncology. Knowledge of disease incidence, outcomes, as well as risk factors is fundamental to all fields involved in research and treatment of patients with brain tumors; thus, epidemiology constitutes an important link between disciplines, indeed the very hub. This was reflected by the scientific program, which included various sessions linking brain tumor epidemiology with clinical neuro-oncology, tissue-based research, and cancer registration. Renowned experts from Europe and the United States contributed their personal perspectives stimulating further group discussions. Several concrete action plans evolved for the group to move forward until next year's meeting, which will be held at the Mayo Clinic at Rochester, MN, USA.


Filaria control and elimination: diagnostic, monitoring and surveillance needs.
Gold standard diagnosis using blood films or skin snips has dimished relevance as mass drug distribution programmes for control of filaria infections expand. The view of 'diagnosis' and its relevance at the individual level has changed, as it has been recognised that the spectrum of programmatic processes (mapping, mass drug interventions, monitoring and evaluation, and surveillance) require different approaches as different questions are asked at each stage. The feasibility and relevance of skin biopsy or blood film examination is challenged when mass drug distribution seeks to treat all eligibles in communities. The need to expand programmes rapidly by identifying the highest risk communities has seen the development of rapid assessment methods, such as rapid epidemiological mapping of onchocerciasis (REMO) and rapid epidemiological assessment (REA) for onchocerciasis, immunochromatographic test (ICT)-based mapping for lymphatic filariasis (LF), and Rapid Assessment Procedure for Loiasis (RAPLOA) for Loa, to reduce the risk of serious adverse events and to guide projects in high-risk communities. As programmes reduce the prevalence through mass drug distribution, more sensitive techniques are required to define endpoints, for LF in particular where the programmatic goal is elimination; for onchocerciasis, sensitive surveillance tools are required particularly in those areas where such risks of recrudescence are high. Whilst much progress has been made in the development and deployment of rapid methods, there are still specific needs for antigen detection in onchocerciasis, whilst standardisation of a panel of tools for LF will allow the definition of endpoint parameters so that countries can decide when mass drug administration (MDA) can be stopped and have a sensitive post-MDA surveillance system.


White matter integrity, hippocampal volume, and cognitive performance of a world-famous nonagenarian track-and-field athlete.
Physical activity (PA) and cardiorespiratory fitness (CRF) are associated with successful brain and cognitive aging. However, little is known about the effects of PA, CRF, and exercise on the brain in the oldest-old. Here we examined white matter (WM) integrity, measured as fractional anisotropy (FA) and WM hyperintensity (WMH) burden, and hippocampal (HIPP) volume of Olga Kotelko (1919-2014). Olga began training for competitions at age of 77 and as of June 2014 held over 30 world records in her age category in track-and-field. We found that Olga's WMH burden was larger and the HIPP was smaller than in the reference sample (58 healthy low-active women 60-78 years old), and her FA was consistently lower in the regions overlapping with WMH. Olga's FA in many normal-appearing WM regions, however, did not differ or was greater than in the reference sample. In particular, FA in her genu corpus callosum was higher than any FA value observed in the reference sample. We speculate that her relatively high FA may be related to both successful aging and the beneficial effects of exercise in old age. In addition, Olga had lower scores on memory, reasoning and speed tasks than the younger reference sample, but outperformed typical adults of age 90-95 on speed and memory. Together, our findings open the possibility of old-age benefits of increasing PA on WM microstructure and cognition despite age-related increase in WMH burden and HIPP shrinkage, and add to the still scarce neuroimaging data of the healthy oldest-old (>90 years) adults.


A micro-scale surface-structured PCL scaffold fabricated by a 3D plotter and a chemical blowing agent.
To study cell responses, polymeric scaffolds with a controllable pore size and porosity have been fabricated using rapid-prototyping methods. However, the scaffolds fabricated by rapid prototyping have very smooth surfaces, which tend to discourage initial cell attachment. Initial cell attachment, migration, differentiation and proliferation are strongly dependent on the chemical and physical characteristics of the scaffold surface. In this study, we propose a three-dimensional (3D) plotting method supplemented with a chemical blowing agent to produce a surface-modified 3D scaffold in which the surface is inscribed with nano- and micro-sized pores. The chemically-blown 3D polymeric scaffold exhibited positive qualities, including the compressive modulus, hydrophilicity and initial cell adhesion. Cell cultures on the scaffolds demonstrated that chondrocytes interacted better with the surface-modified scaffold than with a normal 3D scaffold.


Surface-grafted, environmentally sensitive polymers for biofilm release.
Controlling bacterial biofouling is desirable for almost every human enterprise in which solid surfaces are introduced into nonsterile aqueous environments. One approach that is used to decrease contamination of manufactured devices by microorganisms is using materials that easily slough off accumulated material (i.e., fouling release surfaces). The compounds currently used for this purpose rely on low surface energy to inhibit strong attachment of organisms. In this study, we examined the possible use of environmentally responsive (or "smart") polymers as a new class of fouling release agents; a surface-grafted thermally responsive polymer, poly(N-isopropylacrylamide) (PNIPAAM), was used as a model compound. PNIPAAM is known to have a lower critical solubility temperature of approximately 32 degrees C (i.e., it is insoluble in water at temperatures above 32 degrees C and is soluble at temperatures below 32 degrees C). Under experimental conditions, >90% of cultured microorganisms (Staphylococcus epidermidis, Halomonas marina) and naturally occurring marine microorganisms that attached to grafted PNIPAAM surfaces during 2-, 18-, 36-, and 72-h incubations were removed when the hydration state of the polymer was changed from a wettability that was favorable for attachment to a wettability that was less favorable. Of particular significance is the observation that an organism known to attach in the greatest numbers to hydrophobic substrata (i.e., H. marina) was removed when transition of PNIPAAM to a more hydrated state occurred, whereas an organism that attaches in the greatest numbers to hydrophilic substrata (i.e., S. epidermidis) was removed when the opposite transition occurred. Neither solvated nor desolvated PNIPAAM exhibited intrinsic fouling release properties, indicating that the phase transition was the important factor in removal of organisms. Based on our observations of the behavior of this model system, we suggest that environmentally responsive polymers represent a new approach for controlling biofouling release.


Effect of glucocorticoids on C3 gene expression by the A549 human pulmonary epithelial cell line.
The third component of C, C3, is the key opsonin of the C cascade and is produced locally within the lung by pulmonary epithelial cells, macrophages, and fibroblasts. Because glucocorticoids regulate the maturation and expression of several physiologically important genes in pulmonary epithelial cells, we examined the effects of glucocorticoids on C3 mRNA expression and C3 synthesis by the human pulmonary epithelial cell line, A549. Treatment with dexamethasone enhanced C3 production in a time- and dose-dependent fashion such that concentrations of dexamethasone greater than or equal to 0.001 microM significantly increased C3 production on day 3 of culture. Natural glucocorticoids, corticosterone, cortisol, and 11-deoxycortisol also increased C3 concentrations in A549 supernatants. Both cycloheximide and the glucocorticoid receptor antagonist, RU486, individually inhibited the effect of dexamethasone on C3 production. Northern analysis demonstrated that the steady state 5.2-kb C3 message increased in A549 cells within 10 h of treatment with dexamethasone. RU486 inhibited the effect of dexamethasone on C3 mRNA expression. The integrity of the C3 thiolester bond, as measured by [3H]iodoacetic acid titration and hemolytic assay, was not disrupted by dexamethasone. We conclude that glucocorticoids such as dexamethasone enhance the expression of C3 mRNA and increase the production of functionally active C3 by A549 cells by a mechanism that is mediated by the intracellular glucocorticoid receptor.


Axonin-1, Nr-CAM, and Ng-CAM play different roles in the in vivo guidance of chick commissural neurons.
Immunoglobulin/fibronectin type III-like cell adhesion molecules have been implicated in axon pathfinding based on their expression pattern in the developing nervous system and on their complex interactions described in vitro. The present in vivo study demonstrates that interactions by two of these molecules, axonin-1 on commissural growth cones and Nr-CAM on floor plate cells, are required for accurate pathfinding at the midline. When axonin-1 or Nr-CAM interactions were perturbed, many commissural axons failed to cross the midline and turned instead along the ipsilateral floor plate border. In contrast, though perturbation of Ng-CAM produced a defasciculation of the commissural neurites, it did not affect their guidance across the floor plate.


Optimal fringe angle selection for digital fringe projection technique.
Existing digital fringe projection (DFP) systems mainly use either horizontal or vertical fringe patterns for three-dimensional shape measurement. This paper reveals that these two fringe directions are usually not optimal where the phase change is the largest to a given depth variation. We propose a novel and efficient method to determine the optimal fringe angle by projecting a set of horizontal and vertical fringe patterns onto a step-height object and by further analyzing two resultant phase maps. Experiments demonstrate the existence of the optimal angle and the success of the proposed optimal angle determination method.


In situ X-ray structural studies of a flexible host responding to incremental gas loading.
Crystallographic pressure-lapse snapshots of a porous material responding to gas loading were used to investigate the stepwise uptake of carbon dioxide and acetylene molecules into discrete confined spaces. Based on the data, a qualitative statistical mechanical model was devised that reproduces even subtle features in the experimental gas sorption isotherms.


Prevention and treatment of vitamin D deficiency.
Vitamin D insufficiency and deficiency are widespread in many countries. We review the evidence pertaining to its prevention and treatment. Deficiency may be adequately treated with many different therapeutic regimens of either cholecalciferol or ergocalciferol, owing to the high therapeutic index of both compounds. Nevertheless, the current evidence suggests that regular dosing with oral cholecalciferol (e.g., 60,000 IU weekly) may have slight advantages over other regimens when replenishing vitamin D stores following deficiency. For long-term supplementation, smaller regular doses, such as cholecalciferol 1,000 IU daily, or 10,000 IU weekly, are suitable. Giving reliable and specific advice about appropriate sunlight exposure remains difficult because of differing interindividual skin pigmentation and variable sunlight UVB content at different latitudes, at different times of year, and in different terrestrial environments.


Desferrioxamine elevates pulmonary vascular resistance in humans: potential for involvement of HIF-1.
Hypoxia-inducible factor (HIF)-1 is stabilized by hypoxia and iron chelation. We hypothesized that HIF-1 might be involved in pulmonary vascular regulation and that infusion of desferrioxamine over 8 h would consequently mimic hypoxia and elevate pulmonary vascular resistance. In study A, we characterized the pulmonary vascular response to 4 h of isocapnic hypoxia; in study B, we measured the pulmonary vascular response to 8 h of desferrioxamine infusion. For study A, 11 volunteers undertook two protocols: 1) 4 h of isocapnic hypoxia (end-tidal PO(2) = 50 Torr), followed by 2 h of recovery with isocapnic euoxia (end-tidal PO(2) = 100 Torr), and 2) 6 h of air breathing (control). For study B, nine volunteers undertook two protocols while breathing air: 1) continuous infusion of desferrioxamine (4 g/70 kg) over 8 h and 2) continuous infusion of saline over 8 h (control). In both studies, pulmonary vascular resistance was assessed at 0.5- to 1-h intervals by Doppler echocardiography via the maximum pressure gradient during systole across the tricuspid valve. Results show a progressive rise in pressure gradient over the first 3-4 h with both isocapnic hypoxia (P < 0.001) and desferrioxamine infusion (P < 0.005) to increases of ~16 and 4 Torr, respectively. These results support a role for HIF-regulated gene activation in human hypoxic pulmonary vasoconstriction.


Spectrophotometric determination of sulfide in the presence of sulfite and thiosulfate via the precipitation of bismuth(III) sulfide.
A photometric method has been developed for the determination of sulfide at 10(-5) mol dm(-3) levels, which is based on the reaction of sulfide with a given excess amount of bismuth(III) to form a precipitate of bismuth(III) sulfide and on the spectrophotometric measurement of the residual bismuth(III) at 335 nm after extracting with bismuthiol II reagent from an aqueous solution containing acetate buffer into benzene. The presence of sulfite and thiosulfate up to 0.002 mol dm(-3) did not cause any interference in the determination of sulfide, because both sulfite and thiosulfate do not produce any precipitate with bismuth(III). A linear calibration plot with a negative slope was obtained for sulfide over the range of 5.00 x 10(-7) - 3.00 x 10(-5) mol dm(-3) (16.0 - 960 ppb). An experimental calibration plot was in accord with the theoretical plot, taking into account the known excess of bismuth(III), showing that the reaction of sulfide with bismuth(III) proceeded to completion. The relative standard deviation of results from 10 replicate determinations of standard sulfide (2.00 x 10(-5) mol dm(-3)) was 0.44%. The proposed method was successfully applied to the determination of sulfide in hotspring water samples without any pretreatment.


Anionic sites on the free surface of the peritoneal mesothelium: light and electron microscopic detection using cationic colloidal iron.
To examine charged sites on the peritoneal free surface, cationic and anionic colloidal iron methods were applied. Light microscopy of the human and mouse specimens using cationic colloidal iron staining at pH 1.5-7.3 and successive ferrocyanide treatment resulted in a distinct Prussian blue reaction on the free surface of the parietal, mesenteric and visceral peritonea at all the examined pH values. In transmission electron microscopy of specimens stained with cationic colloidal iron at pH 1.5, colloidal particles accumulated in a dotted fashion on the free surface of the mesothelial cells. At pH 7.3, colloidal particles were deposited as numerous fine strands (100-300 nm in length), whose ends often attached to the luminal aspect of the mesothelial cell membrane. Anionic colloidal iron stain at pH 7.2 gave no deposition of colloidal particles on the mesothelial free surface. Priorly methylated samples lost stainability of the peritoneal free surface to cationic colloidal iron staining at pH 1.5 or 2.5, while methylated-saponified sections recovered it. Pretreatment with neuraminidase inhibited the cationic colloidal iron staining at pH 1.5 on the mesothelial free surface. These results indicate that the mesothelial surface anionic sites ionizing at pH 1.5 are mostly due to the carboxyl group of sialic acid. It is suggested that the peritoneal free surface substance stained with the cationic colloidal iron may be membrane-associated sialomucin, whose rich negative-charged sites may repulse each other to prevent peritoneal adhesion or to maintain peritoneal cavity.


Evidence for a specific influence of the nitrergic pathway on the peripheral pulse waveform in rabbits.
The height of the dicrotic notch between the systolic and diastolic peaks of the peripheral pulse wave, expressed as a fraction of the overall amplitude of the wave, is sensitive to nitric oxide (NO) bioactivity. This phenomenon might form the basis of a simple, non-invasive method for determining endothelial function in vivo. We assessed whether the phenomenon is specific to the NO pathway or whether other vasoactive agents have similar effects. The relative height of the dicrotic notch (RHDN) was determined by photoplethysmography in the rabbit ear. It was dose-dependently decreased by acetylcholine, a stimulator of endothelial NO synthesis, and increased by N(G)-nitro-L-arginine methyl ester (L-NAME), an inhibitor of NO synthesis. There was no effect on RHDN of the alpha-adrenergic blocker phentolamine or the beta-adrenergic blocker propranolol. The cyclo-oxygenase inhibitor indomethacin dose-dependently decreased RHDN but this effect was blocked by L-NAME, suggesting it was mediated by cross-talk with the NO pathway. Changes in RHDN appeared to be independent of heart rate and of the delay between the systolic peak and the notch, but were associated with changes in the slope of the dicrotic limb. Both L-NAME and phentolamine produced multiple diastolic peaks, indicative of wave reflections in the vasculature. These data support the view that changes in RHDN are specific to the NO pathway and provide additional information about the mechanisms involved.


Immunogenicity of a virosomally-formulated Plasmodium falciparum GLURP-MSP3 chimeric protein-based malaria vaccine candidate in comparison to adjuvanted formulations.
In clinical trials, immunopotentiating reconstituted influenza virosomes (IRIVs) have shown great potential as a versatile antigen delivery platform for synthetic peptides derived from Plasmodium falciparum antigens. This study describes the immunogenicity of a virosomally-formulated recombinant fusion protein comprising domains of the two malaria vaccine candidate antigens MSP3 and GLURP. The highly purified recombinant protein GMZ2 was coupled to phosphatidylethanolamine and the conjugates incorporated into the membrane of IRIVs. The immunogenicity of this adjuvant-free virosomal formulation was compared to GMZ2 formulated with the adjuvants Montanide ISA 720 and Alum in three mouse strains with different genetic backgrounds. Intramuscular injections of all three candidate vaccine formulations induced GMZ2-specific antibody responses in all mice tested. In general, the humoral immune response in outbred NMRI mice was stronger than that in inbred BALB/c and C57BL/6 mice. ELISA with the recombinant antigens demonstrated immunodominance of the GLURP component over the MSP3 component. However, compared to the Al(OH)(3)-adjuvanted formulation the two other formulations elicited in NMRI mice a larger proportion of anti-MSP3 antibodies. Analyses of the induced GMZ2-specific IgG subclass profiles showed for all three formulations a predominance of the IgG1 isotype. Immune sera against all three formulations exhibited cross-reactivity with in vitro cultivated blood-stage parasites. Immunofluorescence and immunoblot competition experiments showed that both components of the hybrid protein induced IgG cross-reactive with the corresponding native proteins. A virosomal formulation of the chimeric protein GMZ2 induced P. falciparum blood stage parasite cross-reactive IgG responses specific for both MSP3 and GLURP. GMZ2 thus represents a candidate component suitable for inclusion into a multi-valent virosomal malaria vaccine and influenza virosomes represent a versatile antigen delivery system suitable for adjuvant-free immunization with recombinant proteins.


Universal leukocyte reduction of transfused red cells does not provide benefit to patients undergoing cardiac surgery.
A policy of universal leukocyte reduction of the blood supply in Finland was implemented at the beginning of 2003. The aim of the present evaluation was to determine the potential role of leukocyte-reduced red blood cells in decreasing postoperative infections. A retrospective cohort study. A major university clinic. Consecutive patients undergoing cardiac surgery during the years 2002 and 2003. Transfused patients received either buffy-coat-depleted red blood cells before leukocyte reduction (n = 782) or leukocyte-reduced red blood cells after leukocyte reduction (n = 632). The evaluated outcome parameters were culture-proven postoperative infections, 90-day mortality, and length of stay in the intensive care unit. The percentage of patients transfused with red blood cells (56% v 53%, p = 0.16) and amounts of transfused red blood cells (4.3 +/- 6.7 [3.0] units v 4.3 +/- 6.6 [2.0] units, means +/- standard deviation [median], p = 0.48) were comparable between the study groups (buffy-coat-depleted group and leukocyte-reduced group, respectively). The 90-day mortality (6.6% v 6.3%, p = 0.28), the length of intensive care stay (3.6 +/- 4.7 [2.0] days v 4.3 +/- 7.1 [2.0] days, p = 0.34), and the number of patients with culture-proven infections (8.8% v 10.9%, p = 0.19) were unchanged after universal leukocyte reduction. In multivariate comparisons, the leukocyte reduction was not associated with culture-proven postoperative infections and 90-day mortality. No beneficial effect of the universal leukocyte reduction in cardiac surgery was found for culture-positive infection rates, 90-day mortality, or length of intensive care stay.


Naltrexone in the treatment of alcohol dependence.
Seventy male alcohol-dependent patients participated in a 12-week, double-blind, placebo-controlled trial of naltrexone hydrochloride (50 mg/d) as an adjunct to treatment following alcohol detoxification. Subjects taking naltrexone reported significantly less alcohol craving and days in which any alcohol was consumed. During the 12-week study, only 23% of the naltrexone-treated subjects met the criteria for a relapse, whereas 54.3% of the placebo-treated subjects relapsed. The primary effect of naltrexone was seen in patients who drank any alcohol while attending outpatient treatment. Nineteen (95%) of the 20 placebo-treated patients relapsed after they sampled alcohol, while only eight (50%) of 16 naltrexone-treated patients exposed to alcohol met relapse criteria. Naltrexone was not associated with mood changes or other psychiatric symptoms. Significant side effects (nausea) occurred in two naltrexone-treated subjects, and one naltrexone-treated subject complained of increased pain from arthritis. These results suggest that naltrexone may be a safe and effective adjunct to treatment in alcohol-dependent subjects, particularly in preventing alcohol relapse.


Cholecystokinin cells.
Cholecystokinin (CCK) is an important hormonal regulator of the digestive process. CCK cells are concentrated in the proximal small intestine, and hormone is secreted into the blood upon the ingestion of food. The physiological actions of CCK include stimulation of pancreatic secretion and gallbladder contraction, regulation of gastric emptying, and induction of satiety. Therefore, in a highly coordinated manner, CCK regulates the ingestion, digestion, and absorption of nutrients. CCK is produced by two separate cell types: endocrine cells of the small intestine and various neurons in the gastrointestinal tract and central nervous system. Accordingly, CCK can function as either a hormone or a neuropeptide. This review focuses on the physiology of the CCK cell in the intestine and, in particular, on how the CCK cell is regulated to secrete its hormone product. The effects of ingested nutrients on the CCK cell and the intracellular messenger systems involved in controlling secretion are reviewed. A summary is provided of recent studies examining the electrophysiological properties of CCK cells and newly discovered proteins that act as releasing factors for CCK, which mediate feedback pathways critical for regulated secretion in the intact organism.