Cravings for alcohol in alcohol use disorders are associated with attention deviation to alcohol: An observational study of Japanese in-patients.

Alcohol use disorder (AUD) may be associated with increased attentional bias (AB) toward alcohol-related information. Accordingly, our aim was to explore the relationships between alcohol-related AB, cravings, and risk of relapse among individuals with AUD after treatment. The study group included 24 in-patients with AUD who had completed alcohol withdrawal management. AB was evaluated using an image-based task, with participants asked to select the nonalcoholic image as fast and as accurately as possible and the response time (RT) measured. The intensity of the desire to drink was evaluated using a 100-mm Visual Analog Scale and the risk of relapse using the Alcohol Relapse Risk Scale. Linear regression was used to evaluate the relationship between these variables, with age, gender, duration of hospitalization, and depression score used as covariates. Intensity of cravings was significantly associated with AB RT (R2 =.625) and risk of relapse of drinking behavior (Alcohol Relapse Risk Scale score, R2 =.64). Gender and γ-GTP were significant explanatory variables of identified relationships. The main limitations of our study are a higher proportion of men than women in our study group and the absence of a control group for baseline comparison of AB RTs. This study results suggested that the desire to drink is related to AB among patients with AUD and that the intensity of the desire to drink is related to the risk of a relapse in drinking behavior after AUD treatment.

Correction: The effects of locomotor activity on gastrointestinal symptoms of irritable bowel syndrome among younger people: An observational study.

[This corrects the article DOI: 10.1371/journal.pone.0234089.].

The effects of locomotor activity on gastrointestinal symptoms of irritable bowel syndrome among younger people: An observational study.

Irritable bowel syndrome (IBS) is a common bowel disorder that manifests as unexplained abdominal pain or discomfort and bowel habit changes in the form of diarrhea, constipation, or alternating patterns of the two. Some evidences demonstrate that increased physical activity improves IBS symptoms. Hence, daily exercise is recommended in these patients. In this study, we aimed to investigate the relationship between physical activity and gastrointestinal symptoms in 101 university students (female = 78) with IBS. Participants were examined by Gastrointestinal Symptoms Rating Scale (GSRS), and gait steps were measured for 1 week using a pedometer. The association between the GSRS score and pedometer counts was determined by ordinal logistic modeling analysis. The ordinal logistic regression model for GSRS and locomotor activity showed a significant stepwise fit (z = -3.05, p = 0.002). The logistic curve separated GSRS score of 5 points (moderately severe discomfort) from 2 points (minor discomfort) by locomotor activity. The probability for daily locomotor activity to discriminate between 5 and 4 points of GSRS (i.e., likely to have reverse symptoms) decreased in accordance with increment of steps per day: 78% probability for 4000 steps, 70% probability for 6000 steps, 59% probability for 8000 steps, and 48% probability for 10000 steps. This study demonstrated that the severity of GSRS is associated with the amount of walking in younger people with IBS. These results may be used as a measure to determine the daily step count to reduce the severity of gastrointestinal symptoms in individuals with IBS.

Characterization of embryoid bodies of mouse embryonic stem cells formed under various culture conditions and estimation of differentiation status of such bodies.

Various types of embryoid body (EB) that were formed from mouse embryonic stem (ES) cells under various culture conditions were characterized in terms of gene expression pattern to estimate the differentiation status of the bodies. The gene expression of typical markers (i.e., GATA-4, GATA-6, transthyretin [TTR], alpha-fetoprotein [AFP], Nkx2.5, and alpha-myosin heavy chain [alpha-MHC]) was quantitatively analyzed in various types of EB, and the gene expression pattern of those marker genes was graphically shown for each EB. The gene expression pattern accurately represented the differentiation status of the EBs. The gene expression pattern indicated that the Nkx2.5 and alpha-MHC genes were highly expressed in the EBs formed from 1000 ES cells in a low-adherence 96-well plate. By transferring the EBs into an attachment culture, cardiomyocytes were more efficiently generated in the outgrowth of the EBs. When we increased the seeding cell number from 1000 to 4000 ES cells, the gene expression pattern changed, that is, the expression levels of the TTR and AFP genes increased, whereas those of the Nkx2.5 and alpha-MHC genes decreased, and the trend of differentiation changed from cardiomyogenesis to visceral yolk-sac-like structure formation.

Effect of oxygen on in vitro differentiation of mouse embryonic stem cells.

The monolayer culture of embryonic stem (ES) cells and embryoid body (EB) formation were carried out under various oxygen tensions (i.e., 5% O2, 20% O2, and 40% O2). The ES cells cultured in 5% O2 and 20% O2 exhibited decreased specific alkaline phosphatase (AP) activity with culture time, whereas the ES cells cultured in 40% O2 retained their specific AP activity until culture day 3. The ES cells in 40% O2 maintained an octamer-binding transcription factor 4 (Oct-4; a marker of pluripotent undifferentiated ES cells) gene expression level higher than that at other oxygen tensions. The EB formed in 40% O2 maintained an Oct-4 gene expression level higher than that at other oxygen tensions. The generation of cardiomyocytes and the decline in Oct-4 gene expression level were earliest in the EB formed in 20% O2. It was suggested that the culture condition under a high oxygen tension (40% O2) retards the differentiation of ES cells and a normal oxygen tension (20% O2) allows spontaneous cell differentiation.

A Round-bottom 96-well Polystyrene Plate Coated with 2-methacryloyloxyethyl Phosphorylcholine as an Effective Tool for Embryoid Body Formation.

In this study, we proposed a culture method for forming embryoid bodies (EBs) from mouse embryonic stem (ES) cells using a round-bottom 96-well polystyrene plate coated with 2-methacryloyloxyethyl phosphorylcholine (MPC plate). MPC is a phospholipid biocompatible polymer and prevents cells from adhering to the culture surface. The ES cells were seeded at 1000 cells per well in the MPC plate with 200 mul of medium. After 5 days of static incubation, a spherical cell aggregate termed EB was formed in a well. The size (diameter) of resulting EB was approximately 550 mum and it contained approx. 22,000 cells. It seems that the non-adhesiveness and the roundness of the well are important factors to form a good EB. Transferring the EBs to the attached differentiation culture, the EBs spread out and flattened, and the beating cells (cardiomyocytes) were effectively generated in the outgrowth of EBs. The round-bottom 96-well polystyrene plate coated with MPC is an effective tool for EB formation.

Morphology and albumin secretion of adult rat hepatocytes cultured on a hydrophobic porous expanded polytetrafluoroethylene membrane.

Primary culture of rat hepatocytes was performed on a hydrophobic porous expanded polytetrafluoroethylene (ePTFE) membrane incorporated into the base of a culture dish. Two types of ePTFE membranes, a uniaxially expanded type (ePTFE-1) and a biaxially expanded type (ePTFE-2), could be used as the culture surfaces for hepatocytes. The formation of multicellular aggregates was observed in the culture dish when each membrane type was used. A pore size of 1 mum or higher was adequate for cell adhesion and albumin secretion for both membrane types. The activity of albumin secretion in the dish with the ePTFE membrane was markedly higher than that in the polystyrene dish. Spheroidal multicellular aggregates (spheroids) were observed when hepatocytes were cultured on the ePTFE-1 membrane. The ePTFE-1 membrane maintained the albumin secretion activity for a longer period than the non-expanded PTFE film. It was assumed that the cooperative action of membrane structure and oxygen permeability promoted the formation of cell aggregates and increased the albumin secretion activity.

A simple method for forming embryoid body from mouse embryonic stem cells.

We proposed a simple method for forming an embryoid body (EB) from mouse embryonic stem (ES) cells using a polypropylene 1.5-ml conical tube with a screw cap. An ES cell suspension containing 2 x 10(4) cells was incubated in a conical tube. After 5 d of incubation, a single EB of 440 microm average diameter was formed in the conical tube. The formation efficiency of EB, which is the ratio of the number of tubes showing EB formation to the number of tubes seeded with ES cells, was greater than 99% in the conical tube, while it was approximately 60% in a hanging drop culture. The 5-day-old EB formed by the conical tube method had a sufficient differentiation ability. The beating of the cardiac muscle was microscopically observed in the populations derived from the 5-day-old EB.