Educational technology improves ECG interpretation of acute myocardial infarction among medical students and emergency medicine residents.
Asynchronous online training has become an increasingly popular educational format in the new era of technology-based professional development. We sought to evaluate the impact of an online asynchronous training module on the ability of medical students and emergency medicine (EM) residents to detect electrocardiogram (ECG) abnormalities of an acute myocardial infarction (AMI). We developed an online ECG training and testing module on AMI, with emphasis on recognizing ST elevation myocardial infarction (MI) and early activation of cardiac catheterization resources. Study participants included senior medical students and EM residents at all post-graduate levels rotating in our emergency department (ED). Participants were given a baseline set of ECGs for interpretation. This was followed by a brief interactive online training module on normal ECGs as well as abnormal ECGs representing an acute MI. Participants then underwent a post-test with a set of ECGs in which they had to interpret and decide appropriate intervention including catheterization lab activation. 148 students and 35 EM residents participated in this training in the 2012-2013 academic year. Students and EM residents showed significant improvements in recognizing ECG abnormalities after taking the asynchronous online training module. The mean score on the testing module for students improved from 5.9 (95% CI [5.7-6.1]) to 7.3 (95% CI [7.1-7.5]), with a mean difference of 1.4 (95% CI [1.12-1.68]) (p<0.0001). The mean score for residents improved significantly from 6.5 (95% CI [6.2-6.9]) to 7.8 (95% CI [7.4-8.2]) (p<0.0001). An online interactive module of training improved the ability of medical students and EM residents to correctly recognize the ECG evidence of an acute MI.


Covert fears, anxiety and depression in congenital heart disease.
This study compared anxiety, fears, depression and behavioural problems as occurring in children with congenital heart disease, comparing them with samples of normal children. It further considered the influence of maternal anxiety, as well as analyzing a subgroup of children with cyanotic forms of congenital heart disease to determine if they were at higher risk than acyanotic children for the problems identified. We recruited 40 consecutive children with congenital heart disease without obvious psychosocial problems from the Cardiology clinic at the Alberta Children's Hospital. Of the 40 children, 39 families consented to have the children participate, of which 24 were cyanotic and 15 acyanotic. Children completed the revised versions of the Fear Survey Scale-Revised and the Child Manifest Anxiety Scale as well as the Child Depression Inventory. Mothers completed the Child Behaviour Checklist, and the State Trait Anxiety Scale. Children with congenital heart diseases demonstrated more medical fears, and more physiological anxiety, than the normative samples. More specifically, children with cyanotic forms of congenital heart disease demonstrated more fears of the unknown, physiological anxiety, depression, and delinquent behaviors than the acyanotic children with congenital heart disease. Mothers of the children with cyanotic forms of congenital heart disease scored higher on both the state and trait scales, with higher maternal anxiety correlating with higher anxiety, medical fears and behavioral problems in the child. In a clinical setting, children with congenital heart diseases who do not present with psychological adjustment problems are still at risk for covert physiological anxiety, medical fears, depression and behavioral problems. The children with cyanotic malformations represent a subgroup at higher risk for these problems, which may be further exacerbated by increased maternal anxiety.


Asymptomatic Carotid Stenosis Is Associated With Circadian and Other Variability in Embolus Detection.
Background and Purpose: Variability in transcranial Doppler (TCD) detection of embolic signals (ES) is important for risk stratification. We tested the effect of time of day on ES associated with 60-99% asymptomatic carotid stenosis. Materials and Methods: Subjects were from the Asymptomatic Carotid Stenosis Embolus Detection (ASED) Study such that half were previously ES-positive and half ES-negative with 6-monthly 60-min TCD monitoring. All underwent bilateral TCD monitoring for two 12-h sessions separated by 24 h. ES detection rates were calculated using 6 and 4-h intervals from midnight and effective TCD monitoring time. Results: Ten subjects (8 male, mean age 79.5 years) were monitored. Over 24 h, 5/10 study arteries with 60-99% asymptomatic carotid stenosis were ES-positive (range 1-28 ES/artery, 56 total ES from 177.9 total effective monitoring hours). The remaining five study arteries and all eight successfully monitored contralateral arteries were ES-negative. Using 6-h intervals the mean ES detection rate peaked at 0600-midday (0.64/h) and was lowest 1800-midnight (0.09/h) with an incidence rate ratio of 7.26 (95% CI 2.52-28.64, P ≤ 0.001). Using 4-h intervals the mean ES detection rate peaked at 0800-midday (0.64/h) and was lowest midnight-0400 (0.12/h) with an incidence rate ratio of 5.51 (95% CI 1.78-22.67, P = 0.001). Conclusions: Embolism associated with asymptomatic carotid stenosis shows circadian variation with highest rates 4-6 h before midday. This corresponds with peak circadian incidence of stroke and other vascular complications. These and ASED Study results show that monitoring frequency, duration, and time of day are important in ES detection.


A Review of Consecutive Cases to Identify the Rate of Underlying Osteomyelitis in Patients Undergoing Surgical Treatment of Gangrene of the Forefoot and Impact of Acute Infection on Outcome Following Amputation.
Medical literature offers no clear treatment guidelines when performing amputations for gangrene of the forefoot despite a high percentage that suffer poor outcome due to infection. Gas gangrene and wet gangrene are often preceded by dry stable gangrene. This is a retrospective review of consecutive patients who underwent forefoot amputation and bone biopsy as treatment of forefoot gangrene by a single surgeon. Procedures performed included digital, ray, or transmetatarsal amputation with bone biopsy sent for both culture and histopathologic evaluation. One hundred patients (35 females, 65 males) met inclusion criteria. Mean follow-up was 9.6 months. Mean age was 63.5 years old. Forty-six out of 100 (46%) had elective amputation while 54/100 (54%) were emergent for acute infection. Vascular intervention was performed in 52/100 (52%). Seventy-eight out of 100 (78%) had histopathologic diagnosis of acute osteomyelitis while 82/100 (82%) had positive bone culture. Patients with acute infection had worse outcomes, with higher rates of more proximal amputation and delayed wound healing. We found that 79.7% of patients who underwent forefoot amputation due to gangrene had underlying osteomyelitis. We also found that those with acute infection during the time of amputation had poorer postamputation outcomes such as delayed wound healing, revision surgery, and high rates of more proximal amputation. Therefore, it may imply that earlier amputation of stable gangrene prior to becoming acutely infected may decrease the occurrence of osteomyelitis and avoid some of the preventable postamputation complications. Further studies are warranted.


Dietary macronutrient content, age-specific mortality and lifespan.
Protein and calorie restrictions extend median lifespan in many organisms. However, studies suggest that among-individual variation in the age at death is also affected. Ultimately, both of these outcomes must be caused by effects of nutrition on underlying patterns of age-specific mortality (ASM). Using model life tables, we tested for effects of dietary macronutrients on ASM in mice ( Mus musculus). High concentrations of protein and fat relative to carbohydrates were associated with low life expectancy and high variation in the age at death, a result caused predominantly by high mortality prior to middle age. A lifelong diet comprising the ratio of macronutrients self-selected by mouse (in early adulthood) was associated with low mortality up until middle age, but higher late-life mortality. This pattern results in reasonably high life expectancy, but very low variation in the age at death. Our analyses also indicate that it may be possible to minimize ASM across life by altering the ratio of dietary protein to carbohydrate in the approach to old age. Mortality in early and middle life was minimized at around one-part protein to two-parts carbohydrate, whereas in later life slightly greater than equal parts protein to carbohydrate reduced mortality.


Changes associated with rat heart chromatin during cardiac hypertrophy.
Studies with N-Pyrenemaleimide, a probe which binds to H3 among histones and tryptophan residues of nonhistone proteins and subsequently emits fluorescence, showed that during developing cardiac hypertrophy, conformational changes due to DNA-histone interactions may be maximum at an early stage of hypertrophy (17% hypertrophy) while the changes due to non-histone protein(s) interactions may be maximum at a latter stage (28% hypertrophy). In vitro transcription analysis in isolated nuclei obtained from normal and hypertrophic hearts showed that nuclei obtained from 40% hypertrophic hearts had a maximum incorporation ability than nuclei obtained from 17% or 28% hypertrophic hearts. Analysis of histones and 0.35 M NaCl extractable nuclear proteins by single dimensional polyacrylamide gel electrophoresis did not reveal any changes in these proteins when obtained from normal and different stages of hypertrophic hearts. DNase I sensitivity studies with nuclei obtained from normal and hypertrophic hearts and with reconstituted nuclei, showed that changes in 0.35 M NaCl extractable proteins may contribute to the respective DNase I sensitivity of various hypertrophic nuclei. These studies also indicated that though nuclei obtained from 40% hypertrophic hearts showed maximum incorporation during in vitro transcription, the DNase I sensitivity is maximum only in nuclei obtained from 28% hypertrophic hearts and not from 40% hypertrophic hearts.


Anti-Inflammatory Activity of Marine Ovothiol A in an In Vitro Model of Endothelial Dysfunction Induced by Hyperglycemia.
Chronic hyperglycemia is associated with oxidative stress and vascular inflammation, both leading to endothelial dysfunction and cardiovascular disease that can be weakened by antioxidant/anti-inflammatory molecules in both healthy and diabetic subjects. Among natural molecules, ovothiol A, produced in sea urchin eggs to protect eggs/embryos from the oxidative burst at fertilization and during development, has been receiving increasing interest for its use as an antioxidant. Here, we evaluated the potential antioxidative/anti-inflammatory effect of purified ovothiol A in an in vitro cellular model of hyperglycemia-induced endothelial dysfunction employing human umbilical vein endothelial cells (HUVECs) from women affected by gestational diabetes (GD) and from healthy mothers. Ovothiol A was rapidly taken up by both cellular systems, resulting in increased glutathione values in GD-HUVECs, likely due to the formation of reduced ovothiol A. In tumor necrosis factor-α-stimulated cells, ovothiol A induced a downregulation of adhesion molecule expression and decrease in monocyte-HUVEC interaction. This was associated with a reduction in reactive oxygen and nitrogen species and an increase in nitric oxide bioavailability. These results point to the potential antiatherogenic properties of the natural antioxidant ovothiol A and support its therapeutic potential in pathologies related to cardiovascular diseases associated with oxidative/inflammatory stress and endothelial dysfunction.


Intrahepatic lymphocyte expression of dipeptidyl peptidase I-processed granzyme B and perforin induces hepatocyte expression of serine proteinase inhibitor 6 (Serpinb9/SPI-6).
Human proteinase inhibitor 9 (PI-9/serpinB9) and the murine ortholog, serine proteinase inhibitor 6 (SPI-6/serpinb9) are members of a family of intracellular serine proteinase inhibitors (serpins). PI-9 and SPI-6 expression in immune-privileged cells, APCs, and CTLs protects these cells against the actions of granzyme B, and when expressed in tumor cells or virally infected hepatocytes, confers resistance to killing by CTL and NK cells. The present studies were designed to assess the existence of any correlation between granzyme B activity in intrahepatic lymphocytes and induction of hepatic SPI-6 expression. To this end, SPI-6, PI-9, and serpinB9 homolog expression was examined in response to IFN-alpha treatment and during in vivo adenoviral infection of the liver. SPI-6 mRNA expression increased 10- to 100-fold in the liver after IFN-alpha stimulation and during the course of viral infection, whereas no significant up-regulation of SPI-8 and <5-fold increases in other PI-9/serpinB9 homolog mRNAs was observed. Increased SPI-6 gene expression during viral infection correlated with influxes of NK cells and CTL. Moreover, IFN-alpha-induced up-regulation of hepatocyte SPI-6 mRNA expression was not observed in NK cell-depleted mice. Additional experiments using genetically altered mice either deficient in perforin or unable to process or express granzyme B indicated that SPI-6 is selectively up-regulated in hepatocytes in response to infiltration of the liver by NK cells that express perforin and enzymatically active granzyme B.


Short-term re-epithelialization of the alkali-burned rabbit cornea.
Pfister and associates have shown that treatment with citrate solution reduces the incidence of stromal ulceration after alkali burn of the cornea. In the course of study of the effect of inhibitors of plasminogen activation on corneal re-epithelialization following alkali burns, we noted that when citrate was used in the medication, the healing process proceeded rapidly for three days; however, the denuded area became abruptly larger on the fourth day, and then resumed healing on the fifth day.


Case report: sclerotic skeletal haemangiomatosis presenting with spinal cord compression--CT and MRI findings.
We report the case of a 59-year-old man with skeletal haemangiomatosis who presented with progressive bilateral lower extremity weakness. Computed tomography (CT) and magnetic resonance imaging (MRI) located the causative lesion in the neural arch of the T4 vertebra. CT demonstrated osseous expansion with a mixed lytic and sclerotic pattern. MRI of the lesion showed hypointensity on T1 weighted images, mixed signal intensity on T2 weighted images and moderate contrast enhancement. Similar but less extensive lesions were present in other vertebrae as well as ribs.


[Effects of different cropping patterns on soil enzyme activities and soil microbial community diversity in oasis farmland].
Effects of long-term cropping patterns on the activities of peroxidase, invertase, arylsulfatase, dehydrogenase and protease were investigated in this paper. Four long-term cropping patterns included (1) 10 years continuous cropping of corn, (2) 8 years continuous cropping of wheat followed by 10 years continuous cropping of cotton, (3) 15 years continuous cropping of cotton, and (4) 6 years continuous cropping of cotton followed by 6 years of wheat/sunflower rotation. The responses of soil bacteria, fungi, ammonia oxidizing bacteria (AOB) , and the ammonia oxidizing archaea (AOA) to different copping patterns were analyzed. The results showed that cropping patterns significantly affected the activities of soil peroxidase, arylsulfatase, dehydrogenase and protease, while had no significant effect on soil invertase activity. The cropping patterns significantly influenced the diversity index of AOA, but had no significant influence on that of soil bacteria, fungi and AOB. The community structures of soil fungi and AOB were more sensitive to cropping patterns than soil bacteria and AOA. In conclusion, long-term continuous cropping of cotton decreased the activities of soil enzymes activities and soil microbial diversity in oasis farmland, while crop rotation could alleviate the negative influence.


Calpain activation in apoptosis of ventral spinal cord 4.1 (VSC4.1) motoneurons exposed to glutamate: calpain inhibition provides functional neuroprotection.
Glutamate toxicity has been implicated in cell death in neurodegenerative diseases and injuries. Glutamate-induced Ca2+ influx may mediate activation of calpain, a Ca2+-dependent cysteine protease, which in turn may degrade key cytoskeletal proteins. We investigated glutamate-mediated apoptosis of VSC4.1 motoneurons and functional neuroprotection by calpain inhibition. Exposure of VSC4.1 cells to 10 microM glutamate for 24 hr caused significant increases in intracellular free [Ca2+], as determined by fura-2 assay. Pretreatment of cells with 10 or 25 microM calpeptin (a cell-permeable calpain-specific inhibitor) for 1 hr prevented glutamate-induced Ca2+ influx. Western blot analyses showed an increase in Bax:Bcl-2 ratio, release of cytochrome c from mitochondria, and calpain and caspase-3 activities during apoptosis. Cell morphology, as evaluated by Wright staining, indicated predominantly apoptotic features following glutamate exposure. ApopTag assay further substantiated apoptotic features morphologically as well as biochemically. Our data showed that calpeptin mainly prevented calpain-mediated proteolysis and apoptosis and maintained whole-cell membrane potential, indicating functional neuroprotection. The results imply that calpeptin may serve as a therapeutic agent for preventing motoneuron degeneration, which occurs in amyotrophic lateral sclerosis and spinal cord injury. In this investigation, we also examined glutamate receptor subtypes involved in the initiation of apoptosis in VSC4.1 cells following exposure to glutamate. Our results indicated that the N-methyl-D-aspartate (NMDA) receptors contributed more than alpha-amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid (AMPA) receptors to glutamate-mediated Ca2+ influx and cell death mechanism. Inhibition of the activities of both NMDA and AMPA receptors protected VSC4.1 cells from glutamate toxicity and preserved whole-cell membrane potential.


White Counties Stand Apart: The Primacy of Residential Segregation in COVID-19 and HIV Diagnoses.
Emerging epidemiological data suggest that white Americans have a lower risk of acquiring COVID-19. Although many studies have pointed to the role of systemic racism in COVID-19 racial/ethnic disparities, few studies have examined the contribution of racial segregation. Residential segregation is associated with differing health outcomes by race/ethnicity for various diseases, including HIV. This commentary documents differing HIV and COVID-19 outcomes and service delivery by race/ethnicity and the crucial role of racial segregation. Using publicly available Census data, we divide US counties into quintiles by percentage of non-Hispanic white residents and examine HIV diagnoses and COVID-19 per 100,000 population. HIV diagnoses decrease as the proportion of white residents increase across US counties. COVID-19 diagnoses follow a similar pattern: Counties with the highest proportion of white residents have the fewest cases of COVID-19 irrespective of geographic region or state political party inclination (i.e., red or blue states). Moreover, comparatively fewer COVID-19 diagnoses have occurred in primarily white counties throughout the duration of the US COVID-19 pandemic. Systemic drivers place racial minorities at greater risk for COVID-19 and HIV. Individual-level characteristics (e.g., underlying health conditions for COVID-19 or risk behavior for HIV) do not fully explain excess disease burden in racial minority communities. Corresponding interventions must use structural- and policy-level solutions to address racial and ethnic health disparities.


[Anti-inflammatory and immunosuppressive effects of laser therapy in patients with rheumatoid arthritis].
The clinical and laboratory findings were examined of 10 patients with seropositive rheumatoid arthritis (RA) treated with a first applied technique of intravenous irradiation of the circulating blood with helium-neon laser combined with external irradiation of the inflamed joints. A distinct antiinflammatory and immunosuppressant effect was attained in all the RA patients. In 80% of the test subjects, the rheumatoid blood factor reduced to 1:20 titres. The treatment method did not cause any side effects or complications and shortened the time of the patients' stay at hospital.


Identifying prenatal cannabis exposure and effects of concurrent tobacco exposure on neonatal growth.
Cannabis is the most frequently used illicit drug among pregnant women, but data describing the effects of prenatal cannabis exposure and concurrent nicotine and cannabis exposures on neonatal growth are inconsistent. Testing of meconium, the first neonatal feces, offers objective evidence of prenatal cannabis exposure, but the relative ability of meconium testing and maternal self-report to identify affected neonates remains unclear. Eighty-six pregnant women provided detailed self-reports of daily cannabis and tobacco consumption throughout pregnancy. Cannabinoids and tobacco biomarkers were identified in oral fluid samples collected each trimester and quantified in meconium at birth. Cannabis-using women were significantly more likely to also consume tobacco, and smoked similar numbers of cigarettes as non-cannabis-using tobacco smokers. As pregnancy progressed, fewer women smoked cannabis and those who continued to use cannabis reported smoking a smaller number of cannabis joints, but positive maternal oral fluid tests cast doubt on the veracity of some maternal self-reports. More neonates were identified as cannabis exposed by maternal self-report than meconium analysis, because many women quit cannabis use after the first or second trimester; meconium was more likely to be positive if cannabis use continued into the third trimester. Cannabis exposure was associated with decreased birth weight, reduced length, and smaller head circumference, even after data were controlled for tobacco coexposure. Prenatal cannabis exposure was associated with fetal growth reduction. Meconium testing primarily identifies prenatal cannabis exposure occurring in the third trimester of gestation.


Protein engineering of the archetypal nitroarene dioxygenase of Ralstonia sp. strain U2 for activity on aminonitrotoluenes and dinitrotoluenes through alpha-subunit residues leucine 225, phenylalanine 350, and glycine 407.
Naphthalene dioxygenase (NDO) from Ralstonia sp. strain U2 has not been reported to oxidize nitroaromatic compounds. Here, saturation mutagenesis of NDO at position F350 of the alpha-subunit (NagAc) created variant F350T that produced 3-methyl-4-nitrocatechol from 2,6-dinitrotoluene (26DNT), that released nitrite from 23DNT sixfold faster than wild-type NDO, and that produced 3-amino-4-methyl-5-nitrocatechol and 2-amino-4,6-dinitrobenzyl alcohol from 2-amino-4,6-dinitrotoluene (2A46DNT) (wild-type NDO has no detectable activity on 26DNT and 2A46DNT). DNA shuffling identified the beneficial NagAc mutation G407S, which when combined with the F350T substitution, increased the rate of NDO oxidation of 26DNT, 23DNT, and 2A46DNT threefold relative to variant F350T. DNA shuffling of NDO nagAcAd also generated the NagAc variant G50S/L225R/A269T with an increased rate of 4-amino-2-nitrotoluene (4A2NT; reduction product of 2,4-dinitrotoluene) oxidation; from 4A2NT, this variant produced both the previously uncharacterized oxidation product 4-amino-2-nitrocresol (enhanced 11-fold relative to wild-type NDO) as well as 4-amino-2-nitrobenzyl alcohol (4A2NBA; wild-type NDO does not generate this product). G50S/L225R/A269T also had increased nitrite release from 23DNT (14-fold relative to wild-type NDO) and generated 2,3-dinitrobenzyl alcohol (23DNBA) fourfold relative to wild-type NDO. The importance of position L225 for catalysis was confirmed through saturation mutagenesis; relative to wild-type NDO, NDO variant L225R had 12-fold faster generation of 4-amino-2-nitrocresol and production of 4A2NBA from 4A2NT as well as 24-fold faster generation of nitrite and 15-fold faster generation of 23DNBA from 23DNT. Hence, random mutagenesis discovered two new residues, G407 and L225, that influence the regiospecificity of Rieske non-heme-iron dioxygenases.


Ataxias and cerebellar dysfunction: involvement of synaptic plasticity deficits?
Adaptive processes within cerebellar circuits, such as long-term depression and long-term potentiation at parallel fiber-Purkinje cell synapses, have long been seen as important to cerebellar motor learning, and yet little attention has been given to any possible significance of these processes for cerebellar dysfunction and disease. Several forms of ataxia are caused by mutations in genes encoding for ion channels located at key junctures in pathways that lead to the induction of synaptic plasticity, suggesting that there might be an association between deficits in plasticity and the ataxic phenotype. Herein we explore this possibility and examine the available evidence linking the two together, highlighting specifically the role of P/Q-type calcium channels and their downstream effector small-conductance calcium-sensitive (SK2) potassium channels in the regulation of synaptic gain and intrinsic excitability, and reviewing their connections to ataxia.


Nucleotide variability and linkage disequilibrium patterns at the porcine FABP5 gene.
Fatty acid binding protein 5 (FABP5) is a major positional and physiological candidate gene for the porcine FAT1 QTL on SSC4. Here we characterize the nucleotide polymorphism and haplotype variability of FABP5 and we compare it with that of FABP4, given their close physical location and similar metabolic roles. DNA resequencing of the FABP5 gene region in 29 pigs from 14 breeds and in European and Japanese wild boars revealed 36 polymorphisms in 5.2 kb, and a nucleotide diversity of 0.19%, comparable to values reported in other domestic species but sixfold lower than that previously found for FABP4. Remarkably, both the nucleotide variability and the haplotype structure of FABP5 and FABP4 were dramatically different, and the Hudson-Kreitman-Aguadé test was highly significant. Nevertheless, both genes also had similarities. The neighbour-joining trees of their haplotypes did not show a geographical arrangement for any of the genes. Besides, both genes presented a similar extent and pattern of linkage disequilibrium. Haplotype blocks did not extend for large stretches ( approximately 1 kb in both genes), and the number of tag SNPs required to capture all variability was higher than previously expected. Our findings indicate that FABP4 and FABP5 have undergone different selective or evolutive processes. The fact that haplotype blocks were so small may require us to increase the number of SNPs in prospective whole-genome association studies in the pig.


Gene expression profiles during human CD4+ T cell differentiation.
To develop a comprehensive catalogue of phenotypic and functional parameters of human CD4(+) T cell differentiation stages, we have performed microarray gene expression profiling on subpopulations of human thymocytes and circulating naive CD4(+) T cells, including CD3(-)CD4(+)CD8(-) intrathymic T progenitor cells, CD3(int)CD4(+)CD8(+) 'double positive' thymocytes, CD3(high)CD4(+)CD8(-) 'single positive' thymocytes, CD3(+)CD4(+)CD8(-) CD45RA(+)CD62L(+) naive T cells from cord blood and CD3(+)CD4(+)CD8(-) CD45RA(+)CD62L(+) naive T cells from adult blood. These subpopulations were sort-purified to >98% purity and their expressed RNAs were analyzed on Affymetrix Human Genome U133 arrays. Comparison of gene expression signals between these subpopulations and with early passage fetal thymic stromal cultures identify: (i) transcripts that are preferentially expressed in human CD4(+) T cell subpopulations and not in thymic stromal cells; (ii) major shifts in gene expression as progenitor T cells mature into progeny; (iii) preferential expression of transcripts at the progenitor cell stage with plausible relevance to the regulation of expansion and differentiation of these cells; and (iv) preferential expression of potential markers of recent thymic emigrants in naive-phenotype CD4(+) T cells from cord blood. Further evaluation of these findings may lead to a better definition of human thymopoiesis as well as to improved approaches to monitor and to augment the function of this important organ of T cell production.


Assessment of genetically modified oilseed rape GT73 for renewal authorisation under Regulation (EC) No 1829/2003 (application EFSA-GMO-RX-026/1).
Following the submission of application EFSA-GMO-RX-026/1 under Regulation (EC) No 1829/2003 from Bayer Agriculture BV on behalf of Bayer CropScience LP, the Panel on Genetically Modified Organisms of EFSA was asked to deliver a scientific risk assessment on the data submitted in the context of the renewal of authorisation application for foods and food ingredients containing, consisting of, or produced from oilseed rape GT73 with the exception of isolated seed protein, and feed produced from this GM oilseed rape, excluding cultivation in the EU. The data received in the context of this renewal application contained post-market environmental monitoring reports, a systematic search and evaluation of literature, updated bioinformatic analyses, and a search for additional documents or studies performed by or on behalf of the applicant. The GMO Panel assessed these data for possible new hazards, modified exposure or new scientific uncertainties identified during the authorisation period and not previously assessed in the context of the original application. Under the assumption that the DNA sequences of the events in oilseed rape GT73 considered for renewal are identical to the sequences of the originally assessed event, the GMO Panel concludes that there is no evidence in renewal application EFSA-GMO-RX-026/1 for new hazards, modified exposure or scientific uncertainties that would change the conclusions of the original risk assessment on oilseed rape GT73.


High-fructose drinks affect microRNAs expression differently in lean and obese mice.
High fructose intake from soft drinks and sweets is assumed to have a negative impact on human health. Yet in spite of intensive research, the molecular mechanisms of these effects have not been fully elucidated yet, for example, the effect of high fructose intake could be different in normal and obese individuals. Four groups of mice were used in this study: control groups of lean mice and mice with obesity induced by a high-fat diet, then both of these groups with or without fructose administration in drinks. In plasma of each group, triacylglycerol, cholesterol, free fatty acids, alanine aminotransferase, insulin and adiponectin were measured. The expression levels of selected microRNAs (miRNAs) in plasma, the liver, white adipose tissue, brown adipose tissue and subcutaneous adipose tissue were quantified. In both lean and obese mice, high fructose intake increased cholesterol amount in the liver, up-regulated hepatic miR-27a, down-regulated miR-33a in white adipose tissue and increased plasmatic level of miR-21. The effect of high fructose intake on other miRNAs in the liver, plasma and adipose tissues differed in normal and obese mice. Fructose intake led to hepatic hypercholesterolemia and aberrant expression of several miRNAs participating in lipid metabolism, adipocytes differentiation and nonalcoholic fatty liver disease promotion. The effect of fructose on miRNAs expression differed in normal and obese mice. Nevertheless, plasmatic miR-21, which was induced by fructose in both lean and obese mice, may be considered as a potential biomarker of excessive fructose intake.


Epidemiology and Comorbidities of Psoriasis among Israeli Adolescents: A Large Cross-Sectional Study.
Although psoriasis can develop at any age, the data regarding its characteristics in adolescents are sparse. This study was designed to determine the psoriasis prevalence and its associations with the body mass index (BMI), lipid profile, and comorbidities in adolescents. This was a nationwide population-based cross-sectional retrospective study of adolescents (16-18 years old) evaluated for military service between January 1999 and January 2014. Our database included 887,765 adolescents (57.1% males), of whom 3,112 (0.35%) were diagnosed with psoriasis. During the 15-year study period, the psoriasis prevalence increased by 1.4-fold, from 0.3 to 0.42% (1.25-fold for the males and 1.63-fold for the females). Certain comorbidities, such as contact dermatitis, hyperhidrosis, and arthritis, were significantly associated with psoriasis (odds ratios [ORs] of 2.26, 1.51, and 5.3, respectively). The adolescents with psoriasis had significantly elevated BMI and triglyceride values. We found increased ORs of 1.34 (95% confidence interval [CI] = 1.25-1.56) and 1.56 (95% CI = 1.32-1.83) for the overweight and obese adolescents, respectively, while a lower BMI (<20) had an opposite effect with psoriasis (OR = 0.8). Based on our results, the psoriasis prevalence in Israeli adolescents is rising. Dermatological comorbidities and an increased BMI were associated with psoriasis in these adolescents. A better understanding of the distinctive epidemiological characteristics of juvenile psoriasis may allow for the early detection of comorbidities and improve its management.


Effects of feeding different levels of dietary fiber through the addition of corn stover on nutrient utilization of dairy heifers precision-fed high and low concentrate diets.
The objective of this experiment was to assess the effects of manipulating dietary fiber by replacing corn silage (CS) with lower quality forage as corn stover (CST) when used in high concentrate (HC) and low concentrate (LC) diets for precision-fed dairy heifers. Eight Holstein heifers (335.6 ± 7.41 kg of body weight) were randomly assigned to 2 levels of concentrate: HC (20% forage) and LC (80% forage), and to a forage type sequence [0% of forage as corn stover (CST), 100% corn silage (CS); 20% CST, 80% CS; 40% CST, 60% CS; and 60% CST, 40% CS] within concentrate level administered according to a split-plot, 4 × 4 Latin square design (21-d periods). Heifers fed HC had higher apparent total-tract dry matter digestibility (DMD). Increasing the fiber level by increasing the amount of CST in the diet resulted in a linear decrease of DMD and organic matter digestibility. Heifers fed LC diets had higher neutral detergent fiber (NDF) digestibility and tended to have lower acid detergent fiber (ADF) digestibility than those fed HC diets. Substituting CS with 20% CST resulted in the highest NDF and ADF digestibilities. Digestibility of N was not different, but N retention increased for HC and decreased quadratically for LC diets. Heifers fed HC diets decreased fecal output, and CST linearly increased these parameters. Urine volume tended to be higher for HC-fed heifers, and increasing dietary fiber through CST inclusion tended to decrease urine output. This shift in water excretion resulted in similar total manure output. Total purine derivative excretion did not differ between treatments, but interacted with CST addition, resulting in a linear increase in microbial protein flow to the duodenum in HC-fed heifers and in a linear decrease in LC diets as CST increased. In conclusion, increasing dietary fiber through CST decreased DMD and organic matter digestibility linearly, whereas NDF and ADF digestibility were maximized when 20% CST was added to HC and LC diets. Microbial protein synthesis increased and decreased linearly with CST addition in HC and LC diets, respectively. Retention of N increased and decreased quadratically with CST addition in HC and LC diets, respectively. Total manure excretion was not different between HC or LC diets.


Identification of the Ghrelin and Cannabinoid CB2 Receptor Heteromer Functionality and Marked Upregulation in Striatal Neurons from Offspring of Mice under a High-Fat Diet.
Cannabinoids have been reported as orexigenic, i.e., as promoting food intake that, among others, is controlled by the so-called "hunger" hormone, ghrelin. The aim of this paper was to look for functional and/or molecular interactions between ghrelin GHSR1a and cannabinoid CB2 receptors at the central nervous system (CNS) level. In a heterologous system we identified CB2-GHSR1a receptor complexes with a particular heteromer print consisting of impairment of CB2 receptor/Gi-mediated signaling. The blockade was due to allosteric interactions within the heteromeric complex as it was reverted by antagonists of the GHSR1a receptor. Cannabinoids acting on the CB2 receptor did not affect cytosolic increases of calcium ions induced by ghrelin acting on the GHSR1a receptor. In situ proximity ligation imaging assays confirmed the expression of CB2-GHSR1a receptor complexes in both heterologous cells and primary striatal neurons. We tested heteromer expression in neurons from offspring of high-fat-diet mouse mothers as they have more risk to be obese. Interestingly, there was a marked upregulation of those complexes in striatal neurons from siblings of pregnant female mice under a high-fat diet.


Arylsulfatase A bound to poly(butyl cyanoacrylate) nanoparticles for enzyme replacement therapy--physicochemical evaluation.
Arylsulfatase A (ASA) deficiency is the cause of metachromatic leucodystrophy (MLD), a lysosomal storage disease associated with severe neurological disorders. Poly(butyl cyanoacrylate) (PBCA) nanoparticles overcoated with polysorbate 80 enabled the delivery of several drugs across the blood-brain barrier to the brain suggesting that these nanoparticles also may transport ASA across this barrier. The objective of this research, therefore, was to evaluate the feasibility of loading ASA onto PBCA nanoparticles. A stable ASA-loaded PBCA nanoparticle formulation was developed that could be easily freeze-dried and stored over a period of more than 8 weeks. The maximum loading capacity for this enzyme was -59 microg per 1 mg of PBCA. In the presence of 3% sucrose as a lyoprotector the activity of freeze-dried ASA was found to be 100% recoverable.


Cationic cell-penetrating peptides induce ceramide formation via acid sphingomyelinase: implications for uptake.
Cationic cell-penetrating peptides (CPP) are receiving increasing attention as molecular transporters of membrane-impermeable molecules. Import of cationic CPP occurs both via endocytosis and - at higher peptide concentrations - in an endocytosis-independent manner via localized regions of the plasma membrane. At present, this endocytosis-independent import of cationic CPP is not well understood, but has been shown to be sensitive to various pharmacological inhibitors, suggesting a role of an unidentified enzymatic activity. Here, we demonstrate that the direct translocation of cationic CPP depends on a CPP-induced translocation of acid sphingomyelinase (ASMase) to the outer leaflet of the plasma membrane and ceramide formation. The involvement of ASMase in uptake was confirmed by a pharmacological inhibition of ASMase by imipramine and a subsequent rescue of uptake through external addition of sphingomyelinase, and by using ASMase-deficient cells. We also found that the threshold for direct CPP translocation can be lowered through addition of sphingomyelinase and that sphingomyelinase enhances the translocation of R9 coupled to low-molecular weight cargos, but not high-molecular weight cargos. In conclusion, we show that a previously poorly understood mechanism of cationic CPP import depends on the ASMase-dependent formation of ceramide on the outer leaflet of the plasma membrane. To our knowledge, this is the first illustration that a class of delivery vectors operates through the induction of an enzymatic activity that changes the lipid composition of the plasma membrane.


Nitrogen recycling buffers against ammonia toxicity from skeletal muscle breakdown in hibernating arctic ground squirrels.
Hibernation is a state of extraordinary metabolic plasticity. The pathways of amino acid metabolism as they relate to nitrogen homeostasis in hibernating mammals in vivo are unknown. Here we show, using pulse isotopic tracing, evidence of increased myofibrillar (skeletal muscle) protein breakdown and suppressed whole-body production of metabolites in vivo throughout deep torpor. As whole-body production of metabolites is suppressed, amino acids with nitrogenous side chains accumulate during torpor, while urea cycle intermediates do not. Using 15N stable isotope methodology in arctic ground squirrels (Urocitellus parryii), we provide evidence that free nitrogen is buffered and recycled into essential amino acids, non-essential amino acids and the gamma-glutamyl system during the inter-bout arousal period of hibernation. In the absence of nutrient intake or physical activity, our data illustrate the orchestration of metabolic pathways that sustain the provision of essential and non-essential amino acids and prevent ammonia toxicity during hibernation.


Ten-year experience with an emergency medicine resident research project requirement.
Controversy exists regarding the value and quality of required emergency medicine (EM) resident scholarly projects. To describe the research designs and presentation rate at national scientific meetings and the publication rate of EM resident scholarly projects at a university-based residency program. The authors reviewed the initial ten years (1993-2002) of resident scholarly projects from an EM residency program. Since the inception of the program, a formal research study has been required of all residents for residency graduation. Scholarly projects were reviewed and categorized by study design. Abstracts from the American Academy of Emergency Medicine (AAEM), American College of Emergency Physicians (ACEP), and Society for Academic Emergency Medicine (SAEM) annual meetings were searched to identify projects presented at any of these national meetings. A PubMed search for resident and faculty investigators was performed, and faculty and graduated residents were queried to identify all resident scholarly projects published in peer-reviewed journals. Eighty-seven residents produced 90 scholarly projects. Study designs were prospective data collection, 42 (47%); retrospective chart review, 38 (42%); survey, 5 (6%); animal, 4 (4%); and computer program development, 1 (1%). Of the 80 projects collecting patient data, 72 were conducted at a single center; 6, at two centers; and 2, at five centers each. Of the 42 prospective clinical studies, 27 (64%) were observational and 15 (36%) were interventional. Forty-six (51%) abstracts were presented at national meetings (SAEM, 20; ACEP, 19; AAEM, 3; and other, 4). Thirty-six (40%) of the projects have been published in peer-reviewed journals. Abstract presentation at national meetings (range, 13%-64% of projects per yr) and manuscript publication rates (range, 0-67% of projects per yr) were variable from year to year. Resident scholarly projects at one institution were equally likely to use a prospective or retrospective design, and most were conducted at a single center. More than half of the projects were presented at national research meetings, and more than a third were subsequently developed into manuscripts and published in peer-reviewed journals. When an original research study is required for satisfying the scholarly requirement for EM residency graduation, resident projects can contribute to the EM literature.


Method for analysis of nanoparticle hemolytic properties in vitro.
Hemolysis (destruction of red blood cells) in vivo can lead to anemia, jaundice, and other pathological conditions; therefore the hemolytic potential of all intravenously administered pharmaceuticals must be evaluated. Nanotechnology-derived devices and drug carriers are emerging as alternatives to conventional small-molecule drugs, and in vitro evaluation of their biocompatibility with blood components is a necessary part of early preclinical development. The small size and unique physicochemical properties of nanoparticles may cause their interactions with erythrocytes to differ from those observed for conventional pharmaceuticals and may also cause interference with standardized in vitro tests. Separating true hemolytic responses from the false-positive or false-negative results caused by particle interference is important for correct interpretation of these tests. Here we describe validation of an in vitro assay for the analysis of nanoparticle hemolytic properties and discuss observed nanointerferences with the assay. We propose alternative methods to avoid misleading results from nanoparticles and discuss the potential relevance of nanoparticle in vitro hemolytic properties to in vivo systems.


Molecular Mechanism of ISC Iron-Sulfur Cluster Biogenesis Revealed by High-Resolution Native Mass Spectrometry.
Iron-sulfur (Fe-S) clusters are ubiquitous protein cofactors that are required for many important biological processes including oxidative respiration, nitrogen fixation, and photosynthesis. Biosynthetic pathways assemble Fe-S clusters with different iron-to-sulfur stoichiometries and distribute these clusters to appropriate apoproteins. In the ISC pathway, the pyridoxal 5'-phosphate-dependent cysteine desulfurase enzyme IscS provides sulfur to the scaffold protein IscU, which templates the Fe-S cluster assembly. Despite their functional importance, mechanistic details for cluster synthesis have remained elusive. Recent advances in native mass spectrometry (MS) have allowed proteins to be preserved in native-like structures and support applications in the investigation of protein structure, dynamics, ligand interactions, and the identification of protein-associated intermediates. Here, we prepared samples under anaerobic conditions and then applied native MS to investigate the molecular mechanism for Fe-S cluster synthesis. This approach was validated by the high agreement between native MS and traditional visible circular dichroism spectroscopic assays. Time-dependent native MS experiments revealed potential iron- and sulfur-based intermediates that decay as the [2Fe-2S] cluster signal developed. Additional experiments establish that (i) Zn(II) binding stabilizes IscU and protects the cysteine residues from oxidation, weakens the interactions between IscU and IscS, and inhibits Fe-S cluster biosynthesis; and (ii) Fe(II) ions bind to the IscU active site cysteine residues and another lower affinity binding site and promote the intermolecular sulfur transfer reaction from IscS to IscU. Overall, these results support an iron-first model for Fe-S cluster synthesis and highlight the power of native MS in defining protein-associated intermediates and elucidating mechanistic details of enzymatic processes.


Globalization, Fertility, and Marital Behavior in a Lowest-Low Fertility Setting.
Declines in marriage and fertility rates in many developed countries have fostered research debate and increasing policy attention. Using longitudinal data from the German Socio-Economic Panel, we analyze the effects of exposure to globalization on fertility and marital behavior in Germany, which was a lowest-low fertility setting until recently. We find that exposure to greater import competition from Eastern Europe led to worse labor market outcomes and lower fertility rates. In contrast, workers in industries that benefited from increased exports had better employment prospects and higher fertility. These effects are driven by low-educated individuals, married men, and full-time workers and reflect changes in the likelihood of having any child (the extensive margin). We find evidence of some fertility postponement and significant effects on completed fertility, but we see little evidence of a significant impact on marital behavior. Our results inform the public debate on fertility rates in settings with lowest-low fertility, such as Germany, during the period under investigation.


p23 transplantation antigen mRNA is differentially expressed in human fetal brain.
As gene expression governs development, we attempted to isolate differentially expressed genes in fetal and adult human brain. RNA samples extracted from adult and 18-24-week-old fetal human brain were reverse transcribed, amplified using twenty combinations of 3'-anchored primers and degenerate 5'-primers, and the resulting cDNA fragments separated on denaturing polyacrylamide gels. Thereafter, 45 (H1-H45) differentially displayed cDNA bands were extracted from the gels, amplified by polymerase chain reaction, and used as probes to detect their mRNA by northern blotting. One of these fragments, H8, confirmed on northern blotting to be highly expressed in fetal brain, was cloned and sequenced. This fragment was homologous to wild type p23 human transplantation antigen. This is phylogenetically a well conserved gene and appears to play an important role in cell growth. Even a single point mutation in the mouse gene results in cell destruction secondary to a cytotoxic T-lymphocyte response. Therefore, our finding that normal human fetal brain expresses high levels of wild type p23 transplantation antigen may have importance in maintaining cell growth during human brain development.


Salmonella modulates vesicular traffic by altering phosphoinositide metabolism.
Salmonella enterica, the cause of food poisoning and typhoid fever, induces actin cytoskeleton rearrangements and membrane ruffling to gain access into nonphagocytic cells, where it can replicate and avoid innate immune defenses. Here, we found that SopB, a phosphoinositide phosphatase that is delivered into host cells by a type III secretion system, was essential for the establishment of Salmonella's intracellular replicative niche. SopB mediated the formation of spacious phagosomes following bacterial entry and was responsible for maintaining high levels of phosphatidylinositol-three-phosphate [PtdIns(3)P] in the membrane of the bacteria-containing vacuoles. Absence of SopB caused a significant defect in the maturation of the Salmonella-containing vacuole and impaired bacterial intracellular growth.


Evaluation of tissue eosinophilia in the pouch and afferent limb in patients with restorative proctocolectomy.
Although tissue eosinophilia in mucosal biopsy specimens from the ileal pouch in patients treated with restorative proctocolectomy is frequently seen, its clinical significance has not been investigated. The aim of this study was to assess whether tissue eosinophilia was associated with disease status of ileal pouches. Hematoxylin and eosin slides of pouch and afferent limb biopsy specimens from 106 patients randomly selected from the Pouchitis Database were evaluated by 2 gastrointestinal pathologists. Of 106 patients, 81 had corresponding mucosal biopsy specimens of the afferent limb that were available for review. Tissue eosinophil infiltration was evaluated in a semiquantitative fashion with scores ranging from 0-3. Univariate and multivariate analyses were performed to assess the association between eosinophil scores and demographic, clinical, endoscopic, and histologic features. Multivariate analyses showed that tissue eosinophilia of the pouch and afferent limb was not associated with chronic inflammatory conditions of the pouch and the presence of concurrent autoimmune-mediated disorders. Tissue eosinophil score of the pouch was significantly higher than that in the corresponding afferent limb in the same patient population (P = 0.043). A high tissue eosinophil score in the afferent limb was associated with non-use nonsteroidal antiinflammatory drug use (odds ratio = 3.5; 95% confidence interval [CI]: 1.2, 10.4) and high endoscopic inflammation scores in the afferent limb (odds ratio = 1.6; 95% CI: 1.1, 2.2). Similar associations were not found in pouch biopsy specimens. Tissue eosinophilia in the pouch was more prominent than that in the afferent limb in patients with restorative proctocolectomy. Tissue eosinophilia in the pouch and afferent limb appeared to be associated with different risk factors. These findings suggest that luminal factors in different topographical locations of the pouch may contribute to eosinophil-mediated inflammation at these sites.


Skin changes in internal malignancy.
Internal malignancies are accompanied by various skin changes which may be specific infiltrates or non-specific changes. This study is aimed at determining the frequency of such changes in malignant disease treatment center attendees in India. A study of 300 confirmed cases of internal malignancy at a malignant disease treatment center was undertaken to evaluate these skin changes. Specific infiltrates were confirmed by histopathology. Statistical methods were employed to calculate significance in non-specific lesions by comparing with 300 controls not suffering from internal malignancy. Skin changes were present in 82 (27.3%). Cutaneous metastases were found in 19 (6.3%); non-contiguous in 5 (1.6%); contiguous in 14 (4.3%). Non-specific skin lesions numbered 74 (11.6%) in 52 patients. Statistically significant non-specific skin changes were acquired ichthyosis, herpes zoster and generalized pruritus. Metastases usually occurred late in internal malignancy (17, 5.6%) except in a case each of histiocytic lymphoma and non-Hodgkin's lymphoma (2, 0.7%) where the lesions preceded malignancy by 3 months and 1 month respectively. Contiguous nodules were a marker of relapse after surgery in 3 (1%).


Directed Evolution of a Highly Specific FN3 Monobody to the SH3 Domain of Human Lyn Tyrosine Kinase.
Affinity reagents of high affinity and specificity are very useful for studying the subcellular locations and quantities of individual proteins. To generate high-quality affinity reagents for human Lyn tyrosine kinase, a phage display library of fibronectin type III (FN3) monobodies was affinity selected with a recombinant form of the Lyn SH3 domain. While a highly specific monobody, TA8, was initially isolated, we chose to improve its affinity through directed evolution. A secondary library of 1.2 × 109 variants was constructed and screened by affinity selection, yielding three variants, two of which have affinities of ~ 40 nM, a 130-fold increase over the original TA8 monobody. One of the variants, 2H7, displayed high specificity to the Lyn SH3 domain, as shown by ELISA and probing arrays of 150 SH3 domains. Furthermore, the 2H7 monobody was able to pull down endogenous Lyn from a lysate of Burkitt's lymphoma cells, thereby demonstrating its utility as an affinity reagent for detecting Lyn in a complex biological mixture.


[Modifying effect of environmental chemical factors on the metabolic activation of carcinogens].
Current investigations of modifications of metabolic activation of carcinogens by the environmental chemical factors which are able to induce or inhibit the metabolic activation are reviewed. From the standpoint of ecological oncology the most promising are the following trends: 1) modelling under experimental conditions of integral ecosystems including the complex of living organisms, environmental carcinogens, and factors which modify their biotransformation; 2) study of the imprinting effectors of metabolism modifiers; 3) detection in the environment of modifiers of metabolic activation of carcinogens. The comprehensive oncological-ecological studies of chemicals which influence different pathways of carcinogen metabolism are very important for both primary prevention of cancer and the environmental protection.


Quantitative measurement of red blood cell central pallor and hypochromasia.
A quantitataive definition and techniques of measurement for central pallor of red blood cells are proposed. These are based on high-resolution measurements of absorbance across the center of the cell. Thus, the measurements reflect both variations in cell thickness and hemoglobin concentration. Although contributions of thickness and concentration may differ in individual cells, to a first approximation, a specific cell may be considered as having a similar concentration of hemoglobin throughout, and thus the major contribution to the central pallor is that due to the difference in thickness between the edges of the cell and the center. The definition proposed expresses central pallor as the percentage volume of indentation, comparing the red cell to a disc of uniform absorbance equal to the maximum found at the cell edges. Population distributions of central pallor then provide a basis for quantitation of hypochromasia. The mean and standard deviation of such distributions are proposed as quantitative descriptors. Sample distributions from 27 normal persons, 8 patients with spherocytic anemia and 26 patients with iron deficiency anemia were studied.


Soy isoflavones protect against H₂O₂-induced injury in human umbilical vein endothelial cells.
The aim of this study was to investigate the effects of soy isoflavones on the injury of human umbilical vein endothelial cells induced by H2O2. EVC‑304 cells were preincubated with soy isoflavones for 12 h, and then exposed to 100 µM H2O2 for 1 h. Cell viability was evaluated by a 3‑(4,5‑di‑methylthiazol‑2‑yl) 2,5‑diphenyltetrazolium bromide assay. The apoptosis of EVC‑304 cells was detected by Hoechst 33258 and Annexin‑V/propidium iodide staining. The oxidative stress‑related biochemical parameters were detected and the expression of apoptosis‑related proteins was examined by western blot analysis. The results showed that incubation with soy isoflavones caused a significant increase in the viability of EVC‑304 cells and a decrease in cell apoptosis induced by H2O2. Soy isoflavones also markedly enhanced the activities of superoxide dismutase and glutathione peroxidase, and reduced the level of malondialdehyde. Western blot analysis results show that soy isoflavones can modulate the activation of nuclear factor‑κB and the mitochondria‑mediated apoptosis signaling pathway. The results of this study indicated the potential biological relevance of soy isoflavones in the therapy of cardiovascular diseases.


Cell proliferation and death in the brain of active and hibernating frogs.
'Binomial' cell proliferation and cell death have been studied in only a few non-mammalian vertebrates, such as fish. We thought it of interest to map cell proliferation/apoptosis in the brain of the frog (Rana esculenta L.) as this animal species undergoes, during the annual cycle, physiological events that could be associated with central nervous system damage. Therefore, we compared the active period and the deep underground hibernation of the frog. Using western blot analysis for proliferating cell nuclear antigen (PCNA), we revealed a positive 36 kDa band in all samples and found higher optical density values in the hibernating frogs than in active frogs. In both active and hibernating frogs, we found regional differences in PCNA-immunoreactive cells and terminal transferase dUTP nick-end labelling apoptotic cells in the ventricular zones and parenchyma areas of the main encephalon subdivisions. During the active period of the frogs, the highest concentration of PCNA-immunoreactive cells was found in the ventricle dorsal zone of the cerebral hemispheres but only some of the cells were apoptotic. By contrast, the tectal and cerebellar ventricular zones had a small or medium amount of PCNA-immunoreactive cells, respectively, and a higher number of apoptotic cells. During hibernation, an increased PCNA-immunoreactive cell number was observed in both the brain ventricles and parenchyma compared with active frogs. This increase was primarily evident in the lateral ventricles, a region known to be a proliferation 'hot spot'. Although differences existed among the brain areas, a general increase of apoptotic cell death was found in hibernating frogs, with the highest number of apoptotic cells being detected in the parenchyma of the cerebral hemispheres and optic tectum. In particular, the increased number of apoptotic cells in the hibernating frogs compared with active frogs in the parenchyma of these brain areas occurred when cell proliferation was higher in the corresponding ventricular zones. We suggest that the high number of dying cells found in the parenchymal regions of hibernating frogs might provide the stimulus for the ventricular zones to proliferate. Hibernating frogs could utilize an increased cell proliferation in the brain areas as a neuroprotective strategy to face cell death and the onset of neurological damages. Therefore, the hibernator promises to be a valuable model for studying the mechanisms naturally carried out by the central nervous system in order to adapt itself or survive adverse conditions.


Treating the cognitive deficits of schizophrenia with alpha4beta2 neuronal nicotinic receptor agonists.
Schizophrenic patients exhibit debilitating impairments of intellectual function. Typical and atypical antipsychotic medications are largely ineffective at treating the cognitive deficits of schizophrenia (CDS), and efforts to discover compounds that treat these symptoms are ongoing. Considerable tobacco use in schizophrenic patients, genetic linkage, and receptor binding studies suggest the involvement of nicotinic acetylcholine receptors (nAChRs) in schizophrenia. Neuronal alpha4beta2 nAChRs are widely distributed in the mammalian brain, and are implicated in normal cognitive functioning in animal models. Ligands of various selectivity and potency have been used to study the role of the alpha4beta2 subtype in schizophrenia. For instance, studies in rodents show that alpha4beta2 agonists improve sensory gating, an information processing function that is deficient in schizophrenia. Pharmacological studies in animals also suggest that alpha4beta2 nAChRs are involved in other cognitive domains that are impaired in schizophrenia, including speed of processing, working memory, visual learning and memory, and social cognition. The non-selective nAChR agonist nicotine has been shown to improve CDS in several human clinical studies, and recent trials have been undertaken to evaluate the efficacy of more alpha4beta2 selective compounds. It remains to be determined whether alpha4beta2 agonists will provide greater efficacy than nicotine for CDS or reducing tobacco use in patients. Pre-clinical evidence to date suggests that agonists of the nicotinic alpha4beta2 subtype could be useful in improving cognitive function in schizophrenic patients.


Localization to chromosome 22 of a gene encoding a human minor histocompatibility antigen.
In human allogeneic bone marrow transplantation, graft-vs-host disease and graft rejection can occur even if the patient and donor are genotypically matched by inheritance for HLA. By definition, these allogeneic reactions are due to disparities in minor histocompatibility Ags (minor HAs). Minor HAs are presented to T lymphocytes as peptides bound to HLA molecules, and appear to be encoded by genes throughout the genome. We derived T lymphocyte clones from the PBL of a patient suffering from chronic graft-vs-host disease after bone marrow transplant from his HLA-identical sister. Clones reactive against minor HAs were selected on the basis of reactivity with pretransplant patient cells, and absence of reactivity with donor cells. One clone (MD2) was found to use HLA-B7 as a restricting element. A plasmid vector (pHEBo) containing cDNA encoding the HLA-B7 molecule was transfected into lymphoblastoid cell lines derived from two large families that previously had been saturation mapped for hundreds of polymorphic loci. When clone MD2 was tested against family K1362, it was found to be reactive with three of four grandparents, both parents, and eight of eleven offspring. The same clone was tested with family K1331, with two of three tested grandparents reactive, one of two parents, and nine of eleven offspring. Computer analysis showed that both family segregation patterns linked to an area on the long arm of chromosome 22, localizing the gene encoding this minor HA near the platelet-derived growth factor-beta and IL-2Rbeta genes.


Data reduction and data visualization for automatic diagnosis using gene expression and clinical data.
Accurate diagnoses of specific diseases require, in general, the review of the whole medical history of a patient. Currently, even though many advances have been made for disease monitoring, domain experts are still requested to perform direct analyses in order to get a precise classification, thus implying significant efforts and costs. In this work we present a framework for automated diagnosis based on high-dimensional gene expression and clinical data. Given that high-dimensional data can be difficult to analyze and computationally expensive to process, we first perform data reduction to transform high-dimensional representations of data into a lower dimensional space, yet keeping them meaningful for our purposes. We used then different data visualization techniques to embed complex pieces of information in 2-D images, that are in turn used to perform diagnosis relying on deep learning approaches. Experimental analyses show that the proposed method achieves good performance, featuring a prediction Recall value between 91% and 99%.


The Land-Sea Breeze of the Red Sea: Observations, Simulations, and Relationships to Regional Moisture Transport.
Unique in situ observations of atmospheric conditions over the Red Sea and the coastal Arabian Peninsula are examined to study the dynamics and regional impacts of the local land-sea breeze cycle (LSBC). During a 26-month data record spanning 2008-2011, observed LSBC events occurred year-round, frequently exhibiting cross-shore wind velocities in excess of 8 m/s. Observed onshore and offshore features of both the land- and sea-breeze phases of the cycle are presented, and their seasonal modulation is considered. Weather Research and Forecasting climate downscaling simulations and satellite measurements are used to extend the analysis. In the model, the amplitude of the LSBC is significantly larger in the vicinity of the steeper terrain elements encircling the basin, suggesting an enhancement by the associated slope winds. Observed and simulated conditions also reflected distinct gravity-current characteristics of the intrinsic moist marine air mass during both phases of the LSBC. Specifically, the advance and retreat of marine air mass was directly tied to the development of internal boundary layers onshore and offshore throughout the period of study. Convergence in the lateral moisture flux resulting from this air mass ascending the coastal topography (sea-breeze phase) as well as colliding with air masses from the opposing coastline (land-breeze phase) further resulted in cumulous cloud formation and precipitation.


Medical student patient experiences before and after duty hour regulation and hospitalist support.
With the growth of hospitalist services and the reduction in residency work hours, medical education has changed dramatically. The objective of this study was to examine changes in junior medical student-patient encounters after initiation of residency work hours and implementation of a large hospitalist practice at our academic medical center. Medicine clerkship students from 2002-2007 recorded the number of hospital patients and their principal diagnoses cared for during a 6-week block rotation. Comparisons were made between clerkship experiences among students in 2002-2004 and 2005-2007 for number of patients and diversity of patient diagnoses seen. Data from the 2004-2005 transition period, when teams fluctuated during implementation of the hospitalist service, were excluded. A total of 4,697 patients were seen by students during the two periods, and patient logs for 154 students (3,253 patients in 2002-2004) and 120 students (1,444 patients in 2005-2007) were compared. The mean number of patients directly cared for by students on their junior medicine clerkship dropped from 21 patients (2002-2004) to 12 patients (2005-2007) per student (p < 0.001). Compared to 2002-2004, fewer students from 2005-2007 helped manage patients with chest pain (85.7% vs. 74.2%, p = 0.016), pancreatitis (66.9% vs. 23.3%, p < 0.001), pneumonia (69.5% vs. 54.2%, p = 0.009), gastroenteritis (45.5% vs. 20.8%, p < 0.001), or cellulitis (46.8% vs. 19.2%, p < 0.001). Alternatively, students from 2005-2007 saw more patients with abdominal pain (64.9% vs. 79.2%, p = 0.010), anemia (44.8% vs. 70.8%, p < 0.001), mental status changes (32.5% vs. 51.7%, p = 0.001), failure to thrive (16.2% vs. 53.3%, p < 0.001), and endocrine disorders (including diabetes, thyroid disorders, Addison's, 51.3% vs. 74.2%, p < 0.001). With institutional and residency changes, junior medicine clerkship students had fewer opportunities for direct care of patients and encountered a different mix of patient diagnoses. Increasingly during their junior medicine clerkship, students may not have exposure to basic medical conditions, which may affect their ability to care for future patients.


Measurement of the intramitochondrial volume in hepatocytes without cell disruption and its elevation by hormones and valinomycin.
Methods have been developed to measure the lysophospholipid content and matrix volume of liver cell mitochondria in situ in order to test the hypothesis that these parameters may be important in the hormonal control of mitochondrial function [Armston, Halestrap & Scott (1982) Biochim. Biophys. Acta 681, 429-439]. No change in the labelling of mitochondrial lysophospholipids with [32P]Pi was detected after treatment of liver cells with glucagon, phenylephrine or vasopressin. Incorporation of [32P]Pi into mitochondrial phosphatidylinositol was enhanced by phenylephrine and vasopressin. Mitochondrial volumes were measured using rapid disruption of cells by sonication into 3H2O and [14C]sucrose or without cell disruption using 3H2O and [14C]mannitol. In control cells the two methods gave values of 1.09 and 0.40 microliters/mg of mitochondrial protein respectively, which represent 19 and 7% respectively of the total cell volume measured with 3H2O and inulin [14C]carboxylic acid. Both methods showed that glucagon, phenylephrine and 1 nm-valinomycin produced significant increases (13% and 26% using sucrose and mannitol respectively) in mitochondrial volume. The increase was coincident with the stimulation of gluconeogenesis from L-lactate and pyruvate and of mitochondrial respiratory chain activity. The effects of glucagon and phenylephrine were additive on both mitochondrial volume and respiratory chain activity, but not on gluconeogenesis. Liver cells exposed to gluconeogenic hormones or low concentrations of valinomycin showed a decrease in light scattering at 520 nM correlating with the change in mitochondrial volume but without a change in whole-cell volume. The time course and hormone sensitivity of this response were similar to those for the hormonal stimulation of gluconeogenesis. The light-scattering response to glucagon, phenylephrine and vasopressin, but not to valinomycin, were greatly reduced or abolished in Ca2+-free media.


Vaccination Status and In-hospital Mortality Among Adults With COVID-19 in Jakarta, Indonesia: A Retrospective Hospital-based Cohort Study.
Prospective studies on vaccination status and mortality related to coronavirus disease 2019 (COVID-19) in low-resource settings are still limited. We assessed the association between vaccination status (full, partial, or none) and in-hospital mortality among COVID-19 patients at most hospitals in Jakarta, Indonesia during the Delta predomination wave. We conducted a retrospective cohort study among hospitalized COVID-19 patients who met the study criteria (>18 years old and admitted for inpatient treatment because of laboratory-confirmed severe acute respiratory syndrome coronavirus 2 infection). We linked individual-level data in the hospital admission database with vaccination records. Several socio-demographic and clinical characteristics were also analyzed. A Cox proportional hazards regression model was used to explore the association between vaccination status and in-hospital mortality in this patient group. In total, 40 827 patients were included in this study. Of these, 70% were unvaccinated (n=28 543) and 19.3% (n=7882) died during hospitalization. The mean age of the patients was 49 years (range, 35-59), 53.2% were female, 22.0% had hypertension, and 14.2% were treated in the intensive care unit, and the median hospital length of stay across the group was 9 days. Our study showed that the risk of in-hospital mortality among fully and partially vaccinated patients was lower than among unvaccinated adults (adjusted hazard ratio [aHR], 0.43; 95% confidence interval [CI], 0.40 to 0.47 and aHR, 0.70; 95% CI, 0.64 to 0.77, respectively). Vaccinated patients had fewer severe outcomes among hospitalized adults during the Delta wave in Jakarta. These features should be carefully considered by healthcare professionals in treating adults within this patient group.


Voltage-Induced Switching Dynamics of a Coupled Spin Pair in a Molecular Junction.
Molecular spintronics is made possible by the coupling between electronic configuration and magnetic polarization of the molecules. For control and application of the individual molecular states, it is necessary to both read and write their spin states. Conventionally, this is achieved by means of external magnetic fields or ferromagnetic contacts, which may change the intentional spin state and may present additional challenges when downsizing devices. Here, we predict that coupling magnetic molecules together opens up possibilities for all electrical control of both the molecular spin states as well as the current flow through the system. By tuning between the regimes of ferromagnetic and antiferromagnetic exchange interaction, the current can be at least an order of magnitude enhanced or reduced. The effect is susceptible to the tunnel coupling and molecular level alignment that can be used to achieve current rectification.


Navigating barriers: two-year follow up on recommendations to improve the use of maternal health guidelines in Kosovo.
Although there are a growing number of initiatives aimed at supporting guideline implementation in resource-constrained settings, few studies assess progress on achieving next steps and goals after the initial activities are completed and the initial funding period has ended. The aim of the current study was to conduct a qualitative process evaluation of progress, barriers, facilitators, and proposed solutions to operationalize nine recommendations to prepare Kosovo to implement the 2012 World Health Organization (WHO) prevention and treatment of postpartum haemorrhage guideline. In 2012, we co-created nine recommendations designed to support implementing the WHO's guideline on the prevention and treatment of postpartum haemorrhage in Kosovo. The current study uses a realist evaluation approach to assess activities and progress two years after the recommendations were developed. The study involved conducting qualitative focus groups and one-on-one interviews with participants from the first meeting to evaluate the activities and progress on the nine recommendations. Forty-three participants provided insights into the barriers and opportunities experienced to date and proposed future directions. Although progress has been made towards implementation of a number of the recommendations, scaling up has been limited by barriers, such as lack of awareness, limited resources, and evaluation challenges. Participants proposed addressing these barriers by building within- and between-country partnerships to facilitate guideline implementation. In addition, participants reported less progress on implementing recommendations related to broader cultural changes, which indicates a need for specific and actionable recommendations to operationalize implementation efforts. In the two years since the initial meeting, there has been mixed progress on the recommendations. Based on participant feedback, we refined the recommendations so that they can be operationalized by health care system stakeholders in Kosovo to further support implementation efforts. It is beneficial to share these lessons learned throughout the implementation process to inform next steps in Kosovo and offer ideas for use in other settings.


The maternal childbirth experience more than a decade after delivery.
Maternal satisfaction with the birth experience is multidimensional and influenced by many factors, including mode of delivery. To date, few studies have investigated maternal satisfaction outside of the immediate postpartum period. This study investigated whether differences in satisfaction based on mode of delivery are observed more than a decade after delivery. This was a planned, supplementary analysis of data collected for the Mothers' Outcomes after Delivery study, a longitudinal cohort study of pelvic floor disorders in parous women and their association with mode of delivery. Obstetric and demographic data were obtained through patient surveys and obstetrical chart review. Maternal satisfaction with childbirth experience was assessed via the Salmon questionnaire, administered to Mothers' Outcomes after Delivery study participants >10 years from their first delivery. This validated questionnaire yields 3 scores: fulfillment, distress, and difficulty. These 3 scores were compared by mode of delivery (cesarean prior to labor, cesarean during labor, spontaneous vaginal delivery, and operative vaginal delivery). In addition, the impact of race, age, education level, parity, episiotomy, labor induction, and duration of second stage of labor on maternal satisfaction were examined. Among 576 women, 10.1-17.5 years from delivery, significant differences in satisfaction scores were noted by delivery mode. Salmon scale scores differed between women delivering by cesarean and those delivering vaginally: women delivering vaginally reported greater fulfillment (0.40 [-0.37 to 0.92] vs 0.15 [-0.88 to 0.66], P < .001) and less distress (-0.34 [-0.88 to 0.38] vs 0.20 [-0.70 to 0.93], P < .001) than those who delivered by cesarean. Women who delivered by cesarean prior to labor reported the greatest median fulfillment scores and the lowest median difficulty scores. Median distress scores were lowest among those who delivered by spontaneous vaginal birth. Among women who underwent cesarean delivery, labor induction and prolonged second stage were associated with higher difficulty scores. These factors did not affect satisfaction scores among women who delivered vaginally. Among women who delivered vaginally, operative vaginal delivery was associated with less favorable scores across all 3 scores. Maternal satisfaction with childbirth is influenced by mode of delivery. The birth experience leaves an impression on women more than a decade after delivery.


Examining relations between performance on non-verbal executive function and verbal self-regulation tasks in demographically-diverse populations.
Self-regulation is a widely studied construct, generally assumed to be cognitively supported by executive functions (EFs). There is a lack of clarity and consensus over the roles of specific components of EFs in self-regulation. The current study examines the relations between performance on (a) a self-regulation task (Heads, Toes, Knees Shoulders Task) and (b) two EF tasks (Knox Cube and Beads Tasks) that measure different components of updating: working memory and short-term memory, respectively. We compared 107 8- to 13-year-old children (64 females) across demographically-diverse populations in four low and middle-income countries, including: Tanna, Vanuatu; Keningau, Malaysia; Saltpond, Ghana; and Natal, Brazil. The communities we studied vary in market integration/urbanicity as well as level of access, structure, and quality of schooling. We found that performance on the visuospatial working memory task (Knox Cube) and the visuospatial short-term memory task (Beads) are each independently associated with performance on the self-regulation task, even when controlling for schooling and location effects. These effects were robust across demographically-diverse populations of children in low-and middle-income countries. We conclude that this study found evidence supporting visuospatial working memory and visuospatial short-term memory as distinct cognitive processes which each support the development of self-regulation.


Do three-dimensional modeling and printing technologies have an impact on the surgical success of percutaneous transsacral screw fixation?
This study aims to determine the role of computed tomography (CT)-derived templates, produced by three- dimensional (3D) modeling, image processing and printing technology, in percutaneous transsacral screw fixation and evaluate the effects of their use on surgical success. This prospective study conducted between June 2018 and December 2019 utilized 15 composite pelvis models for transsacral-transiliac screw fixation. For the procedure, modeled templates were utilized for wiring on the left side of the pelvis models, while the conventional method was performed on the right side of the pelvis models. In the computed tomography images acquired after wiring, appropriate wire position was evaluated. The placed wires held the S1 body appropriately in all of the procedures with or without template use. With the template use, the wires were placed appropriately in the surgical bone corridor suitable for the transsacral-transiliac screw fixation in all of the models. However, with the conventional methods, the wires were not placed in the safe surgical bone corridor in four models. The wire deviation angle in the axial plane was significantly lower in the template group (p=0.001), whereas it was not different between the template group and the conventional method group in the coronal plane (p=0.054). The amount of deviation from the ideal wire entry site was significantly reduced in the template group compared to the conventional method group (p=0.001). With the use of 3D modeling and printing technology, CT-derived templates can be produced and utilized for transsacral screw fixation procedures and their use increases surgical success by reducing the surgical margin of error.


Oesophageal pH monitoring in children: how is it perceived by the parents and does the technique change feeding and daily activity?
To determine whether oesophageal pH monitoring in infants induces changes in daily activity and feeding, and with the help of a questionnaire to evaluate how parents perceive pH monitoring. The parents of 100 children, consecutively referred for oesophageal pH monitoring, were asked by one of the authors (A.A.) to respond to a standardized questionnaire immediately after the end of the procedure. Twenty questions were asked, concerning five topics: feeding (4 questions), activity (6 questions), perception of the parents (6 questions), accuracy of the provided information (3 questions), and previous experience with oesophageal pH-metry (1 question). For statistical analysis, every variable was qualified as "no change" and "change" for feeding and activity, as "positive" or "negative" regarding the perception of the test, and as "sufficient" or "insufficient" regarding the information provided. The answers to the questions were also analysed according to the result of the oesophageal pH monitoring (normal vs abnormal), and according to whether or not there was previous experience of oesophageal pH-metry in a first degree relative. Changes in feeding occurred in 28% of cases and changes in activity in 30%, but did not differ according to the oesophageal pH-metry result. Equally, the perception of the parents regarding the test (37% negative), the acceptability of a second pH-metry test (refusal in 52%) and previous experience with oesophageal pH-metry (positive in 25%) were not influenced by the result of the pH monitoring. Parents of infants with abnormal oesophageal pH monitoring were more frequently dissatisfied with the information about the nature of the test provided before the test and also the results than parents of infants with normal results (52% vs 19%, respectively; p < 0.05). According to the perception of the parents, pH monitoring induces changes in feeding and activity in 28% and 30%%, respectively; 63% of the parents considered pH monitoring to be well tolerated. Parents of infants with abnormal pH monitoring results need more attention.


Hemispheric lateralization and top-down regulation of the prefrontal cortex on sequential memory of familiar faces.
Working memory of familiar faces involves the coordination of multiple brain regions in sensory processing, attention and memory, and relies on robust representations in long-term memory. It is not clear how prior knowledge interacts with bottom-up visual processing at different phases of working memory. In this study, we collected functional magnetic resonance imaging (fMRI) data of 40 right-handed participants during the sequential memory task and recognition task of familiar celebrity faces. We observed strong left-lateralized neural activity in the language-processing areas and right-lateralized activity for visual processing in the dorsal stream. Yet, no obvious hemispheric lateralization was found in either face-selective (fusiform gyrus) or memory-specific (hippocampus) areas. Besides, the left lateralization of prefrontal activity and its task-evoked regulation on visual areas boost face memory performance, i.e., faster reaction and higher accuracy. These findings suggest that the top-down prefrontal regulation plays a critical role in the successful memory of familiar faces. Our study provides neural substrates underlying how familiarity boosts face memory by endorsing prior/common knowledge through left-lateralized language network.


Prevention of parenteral nutrition-induced gut hypoplasia by coinfusion of glucagon-like peptide-2.
Maintaining rats on total parenteral nutrition (TPN) for 6 days significantly reduced mass (-34%), protein (-32%), and DNA (-35%) in small intestine and colon (29-37% decrease). Coinfusion of glucagon-like peptide-2 (GLP-2) normalized each of these variables in duodenum, jejunum, and ileum, but not in colon. Histological analysis of tissue revealed normal mucosa thickness and villus height in small intestine of GLP-2-treated rats, whereas nontreated rats maintained on TPN exhibited villus shortening (-30%) and thinning (-23%) of mucosa. These results suggest that hormonal alterations may be more important than an absence of luminal nutrition in TPN-associated mucosal changes. Additionally, GLP-2 normalization of gut mucosa permits accurate assessment of the influence of reversal of hypoplasia on gut barrier function.


Shared decision-making in end-stage renal disease: a protocol for a multi-center study of a communication intervention to improve end-of-life care for dialysis patients.
End-stage renal disease carries a prognosis similar to cancer yet only 20 % of end-stage renal disease patients are referred to hospice. Furthermore, conversations between dialysis team members and patients about end-of-life planning are uncommon. Lack of provider training about how to communicate prognostic data may contribute to the limited number of end-of-life care discussions that take place with this chronically ill population. In this study, we will test the Shared Decision-Making Renal Supportive Care communication intervention to systematically elicit patient and caretaker preferences for end-of-life care so that care concordant with patients' goals can be provided. This multi-center study will deploy an intervention to improve end-of-life communication for hemodialysis patients who are at high risk of death in the ensuing six months. The intervention will be carried out as a prospective cohort with a retrospective cohort serving as the comparison group. Patients will be recruited from 16 dialysis units associated with two large academic centers in Springfield, Massachusetts and Albuquerque, New Mexico. Critical input from patient advisory boards, a stakeholder panel, and initial qualitative analysis of patient and caretaker experiences with advance care planning have informed the communication intervention. Rigorous communication training for hemodialysis social workers and providers will ensure that standardized study procedures are performed at each dialysis unit. Nephrologists and social workers will communicate prognosis and provide advance care planning in face-to-face encounters with patients and families using a social work-centered algorithm. Study outcomes including frequency and timing of hospice referrals, patient and caretaker satisfaction, quality of end-of-life discussions, and quality of death will be assessed over an 18 month period. The Shared Decision-Making Renal Supportive Care Communication intervention intends to improve discussions about prognosis and end-of-life care with end-stage renal disease patients. We anticipate that the intervention will help guide hemodialysis staff and providers to effectively participate in advance care planning for patients and caretakers to establish preferences and goals at the end of life. NCT02405312.


Effects of multispecies probiotic combination on helicobacter pylori infection in vitro.
Probiotic bacteria alleviate many gastrointestinal symptoms, but the current trend of combining bacteria for additional benefit may make their effects more complex. We characterize four probiotics and their combination in terms of pathogen adhesion, barrier function, cell death, and inflammatory response in Helicobacter pylori-infected epithelial cells. H. pylori-infected Caco-2 cells were pretreated with Lactobacillus rhamnosus GG, Lactobacillus rhamnosus Lc705, Propionibacterium freudenreichii subsp. shermanii Js, Bifidobacterium breve Bb99, or all four organisms in combination. We evaluated the adhesion of H. pylori by in situ immunofluorescence; epithelial barrier function by measurement of transepithelial resistance; apoptosis by measurement of caspase 3 activation; cell membrane leakage by measurement of lactate dehydrogenase release; and inflammation by measurement of interleukin-8 (IL-8), IL-10, prostaglandin E(2) (PGE(2)), and leukotriene B(4) (LTB(4)) release. All probiotics inhibited H. pylori adhesion. L. rhamnosus GG, L. rhamnosus Lc705, P. freudenreichii subsp. shermanii Js, and the combination inhibited H. pylori-induced cell membrane leakage. L. rhamnosus GG, L. rhamnosus Lc705, and the combination initially improved epithelial barrier function but increased the H. pylori-induced barrier deterioration after incubation for 24 to 42 h. L. rhamnosus GG, L. rhamnosus Lc705, and P. freudenreichii subsp. shermanii Js inhibited H. pylori-induced IL-8 release, whereas L. rhamnosus GG, L. rhamnosus Lc705, and B. breve Bb99 suppressed PGE(2) release. None of these anti-inflammatory effects persisted when the probiotics were used in combination. The combination thus increased the levels of IL-8, PGE(2), and LTB(4) released from H. pylori-infected epithelial cells. The proinflammatory actions of the individual components dominated the anti-inflammatory effects when the probiotic bacteria were used in combination. Our results stress that the therapeutic response can be optimized if probiotic strains are characterized before they are used in combination.


Synthesis and Antimalarial Activity of New Enantiopure Aminoalcoholpyrrolo[ 1,2-a]quinoxalines.
We prepared a novel series of enantiopure mefloquine analogues with pyrrolo[ 1,2-a]quinoxaline core in order to fight Plasmodium falciparum resistant strain. To observe the influence of pyrrolo[1,2-a]quinoxaline core versus quinoline core on the antimalarial activity. Four enantiopure aminoalcoholpyrrolo[1,2-a]quinoxalines 2 were synthetized via Sharpless asymmetric dihydroxylation reaction in eight steps. Their antimalarial activity was evaluated on two Plasmodium falciparum strains 3D7 and W2 with a SYBR Green I fluorescence-based method and their cytotoxicity was measured on four cell lines HepG2, THP-1, CHO and HFF. IC50 values of the four compounds 2 were close to the micromolar against the two P. falciparum strains. They were more active against P. falciparum strain W2 vs. P. falciparum strain 3D7. (R)- enantiomers were always more active than their (S)-counterpart whatever the strain. Selectivity indexes of compounds 2 were lower than 100. A novel series of enantiopure aminoalcohols with pyrrolo[1,2-a]quinoxaline core were synthesized in eight steps. They displayed IC50 values close to the micromolar against two P. falciparum strains 3D7 and W2. Although, In this series, 2,8-bistrifluoromethylquinoline was a best core than pyrrolo[1,2-a]quinoxaline for an optimal antimalarial activity, the pyrroloquinoxaline 2b showed an interesting antimalarial activity.


Determining the frequency and mechanisms of HIV-1 and HIV-2 RNA copackaging by single-virion analysis.
HIV-1 and HIV-2 are derived from two distinct primate viruses and share only limited sequence identity. Despite this, HIV-1 and HIV-2 Gag polyproteins can coassemble into the same particle and their genomes can undergo recombination, albeit at an extremely low frequency, implying that HIV-1 and HIV-2 RNA can be copackaged into the same particle. To determine the frequency of HIV-1 and HIV-2 RNA copackaging and to dissect the mechanisms that allow the heterologous RNA copackaging, we directly visualized the RNA content of each particle by using RNA-binding proteins tagged with fluorescent proteins to label the viral genomes. We found that when HIV-1 and HIV-2 RNA are present in viral particles at similar ratios, ∼10% of the viral particles encapsidate both HIV-1 and HIV-2 RNAs. Furthermore, heterologous RNA copackaging can be promoted by mutating the 6-nucleotide (6-nt) dimer initiation signal (DIS) to discourage RNA homodimerization or to encourage RNA heterodimerization, indicating that HIV-1 and HIV-2 RNA can heterodimerize prior to packaging using the DIS sequences. We also observed that the coassembly of HIV-1 and HIV-2 Gag proteins is not required for the heterologous RNA copackaging; HIV-1 Gag proteins are capable of mediating HIV-1 and HIV-2 RNA copackaging. These results define the cis- and trans-acting elements required for and affecting the heterologous RNA copackaging, a prerequisite for the generation of chimeric viruses by recombination, and also shed light on the mechanisms of RNA-Gag recognition essential for RNA encapsidation.


The target region focused imaging method for scanning ion conductance microscopy.
Scanning ion conductance microscopy (SICM) has developed rapidly and has wide applications in biomedicine, single-cell science and other fields. SICM scanning speed is limited by the conventional raster-type scanning method, which spends most of time on imaging the substrate and does not focus enough on the target area. In order to solve this problem, a target region focused (TRF) method is proposed, which can effectively avoid the scanning of unnecessary substrate areas and enables SICM to image the target area only to achieve high-speed and effective local scanning. TRF method and conventional hopping mode scanning method are compared in the experiments using breast cancer cells and rat basophilic leukemia cells as experimental materials. It was demonstrated that our method can reduce the scanning time for a single sample image significantly without losing scanning information or compromising the quality of imaging. The TRF method developed in this paper can provide an efficient and fast scanning strategy for improving the imaging performance of SICM systems, which can be applied to the dynamic features of cell samples in the fields of biology and pharmacology analysis.


A review of intraexaminer and interexaminer reliability of static spinal palpation: a literature synthesis.
The purposes of this study were to locate articles that assessed the reliability of static palpation of the spine and sacroiliac joints, to appraise the quality of these studies, and synthesize their results. A structured literature search was conducted of chiropractic and medical databases PubMed, Manual Alternative and Natural Therapy System, Index to Chiropractic Literature, and Cumulative Index to Nursing and Allied Health Literature from 1965 through October 2007. Reference sections were inspected for additional citations. Only peer-reviewed articles in English containing information about static palpation of the spine or sacroiliac joints were selected. The resulting studies were appraised for quality by both of the authors using a 6-point scale instrument developed to assess the quality of reproducibility studies. The search generated 343 citations, and another 7 were harvested from the reference lists. After removing articles not meeting the inclusion criteria, 29 were retained. A total of 14 studies focused on the reliability of locating painful or tender points, 10 on the location of landmarks, and 5 on position or alignment of bone structures. A higher proportion of studies that assessed painful or tender points reported acceptable levels of reliability. However, there were no significant differences between methods of palpation when considering the proportions of high-quality studies that reported good reliability. Thus, no form of static palpation could be considered to be superior. Reported indices of agreement were generally low. More of the pain palpation studies reported acceptable kappa levels, although no one method of palpation could be deemed clearly superior.