Clinical effects of combined Lactobacillus paracasei and kestose on canine atopic dermatitis.

Probiotics and prebiotics are viable bacteria with beneficial effects on the host and components that selectively act on the beneficial commensal bacteria, respectively. The combined use of probiotics and prebiotics is termed synbiotics. Probiotic intake improves dysbiosis in the intestinal microbiota and can positively affect canine atopic dermatitis (CAD). However, clinical studies on improvements in CAD using synbiotics remain limited. In this study, 15 dogs with CAD who received prednisolone, a synthetic glucocorticoid (GC) used in the treatment of CAD, for more than 90 days were continuously treated with Lactobacillus paracasei M-1 from fermented food as a probiotic, and trisaccharide kestose as a prebiotic, for 90 days to determine their synbiotic effects on CAD. The CAD symptoms were evaluated using the canine atopic dermatitis lesion index (CADLI) and pruritus visual analog scores (PVAS) at 30, 60 and 90 days after synbiotic administration. The total prednisolone use for 90 days pre- and post-administration was also evaluated. Synbiotic administration significantly reduced the CADLI (pre: median, 28.0 [22.0-32.0]; 30 days: median, 20.0 [20.0-28.0]; 60 days: median, 20.0 [10.0-21.0]; 90 days: median, 12.0 [10.0-19.0]) and PVAS (pre: median, 6.0 [5.0-7.0]; 30 days: median, 3.0 [3.0-3.5]; 60 days: median, 3.0 [3.0-3.5]; 90 days: median, 2.0 [2.0-3.5]) scores, and reduced the total prednisone use over 90 days (pre: 112.0 [25-450] mg; post: 80.0 [18.-300.0] mg; p⟨0.001) in the 15 dogs. Thus, the synbiotic activity of L. paracasei M-1 and trisaccharide kestose can improve CAD.

Non-invasive classification of single and double-yolk eggs using Vis-NIR spectroscopy and multivariate analysis.

1. This study was conducted to develop an efficient technique for separating double-yolked (DY) from single-yolked (SY) light brown broiler eggs with comparable shape and size, that were hard to distinguish merely by their external characteristics, using Vis-NIR transmission spectroscopy combined with multivariate analysis.2. Spectroscopic transmission (200-900 nm) was measured after collecting the eggs, and the yolk number was verified by breaking the eggs after boiling. The absorbance of important spectral wavelengths sensitive to yolk amount were identified using feature selection techniques (Principal Component Analysis and Genetic Algorithm).3. Discriminant analysis (DA) and support vector machine (SVM) classifiers were used to develop classification models for DY and SY eggs using the selected important spectral wavelengths.4. When compared to alternative nonlinear techniques, the developed model applying linear discriminant analysis produced greater accuracies in the first (96%) and second (100%) experiments, implying lower inter-egg variability from spectral data and a linear relationship between classes. However, the position and orientation of yolks in DY eggs may limit the classification accuracy of the eggs.

DNA damage modeled with Geant4-DNA: effects of plasmid DNA conformation and experimental conditions.

The chemical stage of the Monte Carlo track-structure (MCTS) code Geant4-DNA was extended for its use in DNA strand break (SB) simulations and compared against published experimental data. Geant4-DNA simulations were performed using pUC19 plasmids (2686 base pairs) in a buffered solution of DMSO irradiated by60Co or137Csγ-rays. A comprehensive evaluation of SSB yields was performed considering DMSO, DNA concentration, dose and plasmid supercoiling. The latter was measured using the super helix density value used in a Brownian dynamics plasmid generation algorithm. The Geant4-DNA implementation of the independent reaction times method (IRT), developed to simulate the reaction kinetics of radiochemical species, allowed to score the fraction of supercoiled, relaxed and linearized plasmid fractions as a function of the absorbed dose. The percentage of the number of SB after •OH + DNA and H• + DNA reactions, referred as SSB efficiency, obtained using MCTS were 13.77% and 0.74% respectively. This is in reasonable agreement with published values of 12% and 0.8%. The SSB yields as a function of DMSO concentration, DNA concentration and super helix density recreated the expected published experimental behaviors within 5%, one standard deviation. The dose response of SSB and DSB yields agreed with published measurements within 5%, one standard deviation. We demonstrated that the developed extension of IRT in Geant4-DNA, facilitated the reproduction of experimental conditions. Furthermore, its calculations were strongly in agreement with experimental data. These two facts will facilitate the use of this extension in future radiobiological applications, aiding the study of DNA damage mechanisms with a high level of detail.

TOPAS-nBio validation for simulating water radiolysis and DNA damage under low-LET irradiation.

The chemical stage of the Monte Carlo track-structure simulation code Geant4-DNA has been revised and validated. The root-mean-square (RMS) empirical parameter that dictates the displacement of water molecules after an ionization and excitation event in Geant4-DNA has been shortened to better fit experimental data. The pre-defined dissociation channels and branching ratios were not modified, but the reaction rate coefficients for simulating the chemical stage of water radiolysis were updated. The evaluation of Geant4-DNA was accomplished with TOPAS-nBio. For that, we compared predicted time-dependentGvalues in pure liquid water for·OH, e-aq, and H2with published experimental data. For H2O2and H·, simulation of added scavengers at different concentrations resulted in better agreement with measurements. In addition, DNA geometry information was integrated with chemistry simulation in TOPAS-nBio to realize reactions between radiolytic chemical species and DNA. This was used in the estimation of the yield of single-strand breaks (SSB) induced by137Csγ-ray radiolysis of supercoiled pUC18 plasmids dissolved in aerated solutions containing DMSO. The efficiency of SSB induction by reaction between radiolytic species and DNA used in the simulation was chosen to provide the best agreement with published measurements. An RMS displacement of 1.24 nm provided agreement with measured data within experimental uncertainties for time-dependentGvalues and under the presence of scavengers. SSB efficiencies of 24% and 0.5% for·OH and H·, respectively, led to an overall agreement of TOPAS-nBio results within experimental uncertainties. The efficiencies obtained agreed with values obtained with published non-homogeneous kinetic model and step-by-step Monte Carlo simulations but disagreed by 12% with published direct measurements. Improvement of the spatial resolution of the DNA damage model might mitigate such disagreement. In conclusion, with these improvements, Geant4-DNA/TOPAS-nBio provides a fast, accurate, and user-friendly tool for simulating DNA damage under low linear energy transfer irradiation.

Income-based inequality in nationwide general health checkup participation in Japan.

OBJECTIVE: In Japan, it is mandatory for employers to provide general health checkup opportunities to employees. Although many companies have subsidized checkups for employees' dependent family members, their participation is low. We assessed income-based inequality in the participation of employees' dependents in the general health checkup. STUDY DESIGN: This is a cross-sectional descriptive study. Annual participation rate in general health checkup and various factors including income, age, and sex were collected and analyzed to examine the income-based inequality of participation rate in general health checkup. METHODS: The data for the present study were sourced from the Fukuoka Branch of the Japan Health Insurance Association, a large medical insurer in Japan. We extracted data of 196,057 dependents aged 40-74 years. We conducted a multiple logistic regression analysis using participation from April 2015 to March 2016 as dependent variable and income category ranging from 1 (lowest) to 4 (highest) between April and June 2015 as independent variable (adjusted for sex and age). We computed slope index of inequality (SII) and relative index of inequality (RII) based on income category. RESULTS: Higher the income, the more likely were dependents to participate in the general health checkup. SII for the participation rate of general health checkup ranged between -0.02 (95% confidence interval [CI]: -0.07 to 0.03) and 0.06 (0.03-0.09) for men; 0.03 (0.01-0.06) and 0.10 (0.09-0.11) for women. RII for the participation rate of general health checkup ranged between -0.19 (95% CI: -0.66 to 0.29) and 0.88 (0.15-1.61) for men; 0.22 (0.05-0.39) and 0.68 (0.60-0.76) for women. The highest inequality existed for men in their 50s and 60s and women in their 50s; the lowest inequality was among men and women aged 70-74 years. CONCLUSION: There was income-based inequality in participation in the general health checkup among dependents (family members) of the insured persons. The degree of inequality differed with age group. It cannot be explained solely by financial barrier among low-income group, rather it may reflect Japanese unique context in medical insurance system.

Non-destructive sex-specific monitoring of early embryonic growth rate in light brown broiler eggs using light transmission and its correlation with hatching time and chick weight.

1. Monitoring early embryonic growth rate (EGR) has significant economic and animal welfare benefits. This study focuses on monitoring sex-specific early EGR using light transmission, and correlating this with hatching time and chick weight. For broiler eggs in particular, spectral masking of the light brown eggshells needed to be addressed. This was done using longitudinal visible transmission spectroscopy combined with eggshell colour image analysis. 2. Prior to incubation, colour images of eggs were captured followed by daily measurements of transmission spectra of eggs from days one to nine of incubation. The sex of the eggs was subsequently verified 2 d after hatching. 3. To accurately and sensitively determine sex differences in EGR using light transmission, while minimising interference from eggshell colour and thickness, the ratio of longitudinal transmissions was determined to be most effective at 575 and 610 nm. 3. Embryonic growth was detectable from d 3 (72 h) of incubation, 24 h earlier than previously reported lateral transmission measurements. However, at this time, low blood levels meant that no significant sex-differences (P > 0.05) for the mean T575/T610 ratio were detectable. This may have been due, in part, to spectral masking from the light brown eggshells. At d 7, female embryos had a significantly lower (P < 0.05) mean T575/T610 ratio than males. 4. Although the T575/T610 ratio had low correlations with hatching time and hatch-weight of chicks, this could be a good starting point for further non-destructive investigations for such predictions. 5. In conclusion, the methodology had the sensitivity to differentiate sex-specific early EGR in broiler eggs, even with pigmented eggshells, and has the potential to advance precision hatchery management and poultry research.

Cumulative incidence of femoral localized periosteal thickening (beaking) preceding atypical femoral fractures in patients with rheumatoid arthritis.

The incidence of localized periosteal thickening (LPT, also termed beaking) of the lateral cortex that often precedes an atypical femoral fracture (AFF) was not high in patients with rheumatoid arthritis (RA) but incomplete AFFs developed in two patients. Higher-dose prednisolone was a significant risk factor for LPT in patients with RA. INTRODUCTION: Atypical femoral fractures (AFFs) are stress fractures; bisphosphonate (BP) use is a major risk factor for the development of such fractures. Localized periosteal thickening (LPT, also termed beaking) of the lateral cortex often precedes a complete or incomplete AFF. We evaluated the incidence of latent LPT in patients with rheumatoid arthritis (RA), to evaluate LPT progression, and to define LPT risk factors. METHODS: A total of 254 patients with RA were included; all underwent annual X-ray evaluation, dual-energy X-ray absorptiometry, and analyses of serum and bone metabolic markers for 2-3 years. LPT of the lateral cortex was sought in femoral X-rays. RESULTS: The incidence of LPT was 2.4% (6/254). Among patients on both BP and prednisolone (PSL) at enrollment, the incidence was 2.3% (3/131). Two femurs of two patients with LPT developed incomplete AFFs; LPT was extensive and associated with endosteal thickening. One patient had been on BP and PSL and microscopic polyangiitis was comorbidity. The other was on a selective estrogen receptor modulator and PSL. A daily PSL dose >5 mg (OR 11.4; 95%CI 2.15-60.2; p = 0.004) and higher-dose methotrexate (OR 1.22; 95%CI 1.01-1.49; p = 0.043) were significant risk factors for LPT. CONCLUSIONS: The incidence of latent LPT was not high (2.4%) but incomplete AFFs developed in two RA patients. Higher-dose PSL because of a comorbid disease requiring glucocorticoid treatment other than RA or refractory RA were risk factors for LPT; X-ray screening for latent LPT would usefully prevent complete AFFs.

LET-Dependent Intertrack Yields in Proton Irradiation at Ultra-High Dose Rates Relevant for FLASH Therapy.

FLASH radiotherapy delivers a high dose (≥10 Gy) at a high rate (≥40 Gy/s). In this way, particles are delivered in pulses as short as a few nanoseconds. At that rate, intertrack reactions between chemical species produced within the same pulse may affect the heterogeneous chemistry stage of water radiolysis. This stochastic process suits the capabilities of the Monte Carlo method, which can model intertrack effects to aid in radiobiology research, including the design and interpretation of experiments. In this work, the TOPAS-nBio Monte Carlo track-structure code was expanded to allow simulations of intertrack effects in the chemical stage of water radiolysis. Simulation of the behavior of radiolytic yields over a long period of time (up to 50 s) was verified by simulating radiolysis in a Fricke dosimeter irradiated by 60Co γ rays. In addition, LET-dependent G values of protons delivered in single squared pulses of widths, 1 ns, 1 µs and 10 µs, were obtained and compared to simulations using no intertrack considerations. The Fricke simulation for the calculated G value of Fe3+ ion at 50 s was within 0.4% of the accepted value from ICRU Report 34. For LET-dependent G values at the end of the chemical stage, intertrack effects were significant at LET values below 2 keV/µm. Above 2 keV/µm the reaction kinetics remained limited locally within each track and thus, effects of intertrack reactions remained low. Therefore, when track structure simulations are used to investigate the biological damage of FLASH irradiation, these intertrack reactions should be considered. The TOPAS-nBio framework with the expansion to intertrack chemistry simulation provides a useful tool to assist in this task.

Nondestructive sex-specific monitoring of early embryonic development rate in white layer chicken eggs using visible light transmission.

1. Sex-specific variations in early embryonic development rates may pre-empt later variations in embryonic development through to pipping and hatching. Given that erythropoiesis (blood production) can be equated with early embryonic growth rate, it was hypothesised that blood pigment haemoglobin can act as a specific spectral fingerprint for changes in growth rate. Moreover, by measuring longitudinal, rather than lateral, spectral transmission through the egg, a more consistent spectrum with a higher signal-to-noise ratio could be captured.2. Longitudinal visible transmission (T575/T598 ratio), which is sensitive to haemoglobin, was used to monitor sex-specific early embryonic development rate in white layer chicken eggs from d 0 to 8 of incubation. The sex of these eggs was subsequently confirmed two days after hatching.3. Embryonic development was detectable from d 3 (72 h) of incubation, 36 h earlier than previously reported lateral spectral measurements, supporting the greater sensitivity of longitudinal measurements.4. At d 3, the mean T575/T598 ratio for male embryos was significantly lower (P < 0.001) (i.e. higher absorbance of haemoglobin) than for female embryos, which was thought to be due to sex-differences in early embryogenesis. On the other hand, female embryos had a significantly lower (P < 0.05) mean T575/T598 ratio than male embryos at d 7 of incubation, presumably due to the combined effects of oestrogen synthesis receptors and enzymes on erythropoiesis in female embryos at this time.5. In conclusion, the proposed methodology has the sensitivity to differentiate sex-specific embryonic development rates during early incubation and the potentiality to advance precision incubation management and poultry research.

Identifying Frail-Related Biomarkers among Community-Dwelling Older Adults in Japan: A Research Example from the Japanese Gerontological Evaluation Study.

We examined correlating clinical biomarkers for the physical aspect of frailty among community-dwelling older adults in Japan, using Japanese Gerontological Evaluation Study (JAGES). We used information from the JAGES participants (N = 3,128) who also participated in the community health screening in 2010. We grouped participants' response to the Study of Osteoporotic Fracture (SOF) Frailty Index into robust (=0), intermediate frail (=1), and frail (=2+) ones to indicate physical aspect of frailty. Independent of sex and age, results from multinomial logistic regression showed above normal albumin and below normal HDL and haemoglobin levels were positively associated with intermediate frail (RRR = 1.99, 95% CI = 1.22-3.23; RRR = 1.36, 95% CI = 1.33-1.39; RRR = 1.36, 95% CI = 1.23-1.51, resp.) and frail cases (RRR = 2.27, 95% CI = 1.91-2.70; RRR = 1.59, 95% CI = 1.51-1.68; RRR = 1.40, 95% CI = 1.28-1.52, resp.). Limited to women, above normal Hb1Ac level was similarly associated with intermediate frail and frail cases (RRR = 1.18, 95% CI = 1.02, 1.38; RRR = 2.56, 95% CI = 2.23-2.95, resp.). Use of relevant clinical biomarkers can help in assessment of older adults' physical aspect of frailty.

Japan health system review

The Asia Pacific Observatory on Health Systems and Policies (the APO) is a collaborative partnership of interested governments, international agencies, foundations, and researchers that promotes evidence-informed health systems policy regionally and in all countries in the Asia Pacific region. The APO collaboratively identifies priority health system issues across the Asia Pacific region; develops and synthesizes relevant research to support and inform countries' evidence-based policy development; and builds country and regional health systems research and evidence-informed policy capacity.

High and pointed type of femoral localized reaction frequently extends to complete and incomplete atypical femoral fracture in patients with autoimmune diseases on long-term glucocorticoids and bisphosphonates.

Once a localized reaction (beaking) was detected, discontinuation of bisphosphonates (BPs) and switching to vitamin D supplementation or teriparatide therapy effectively improved its shape. When the localized reaction was high, of the pointed type, and/or accompanied by prodromal pain, the risks of complete and incomplete atypical femoral fracture increased and consideration of prophylactic fixation for such patients was required. INTRODUCTION: Femoral localized reaction (localized periosteal thickening of the lateral cortex, beaking) is reported to precede atypical femoral fractures (AFFs) and to develop in 8-10% of patients with autoimmune diseases taking BPs and glucocorticoids. The aims of the present study were to retrospectively investigate the shapes of localized reaction to consider how to manage the condition. METHODS: Twenty femora of 12 patients with autoimmune diseases who were on BPs and glucocorticoids exhibited femoral localized reaction. The heights of localized reaction were measured and the shapes classified as pointed, arched, and other. Localized reaction changes were divided into three categories: deterioration, no change, and improvement. A severe form of localized reaction was defined; this was associated with prodromal pain, de novo complete AFF, or incomplete AFF with a fracture line at the localized reaction. RESULTS: The mean height of localized reaction was 2.3 ± 0.8 mm (range, 1.0-3.7 mm) and the pointed type was 35%. Localized reaction was significantly higher (3.3 ± 0.8 vs. 2.1 ± 0.7 mm; p = 0.003) and the pointed type more common (78 vs. 27%; p = 0.035) in those with the severe form of localized reaction. Seven patients with localized reactions discontinued BPs just after localized reaction was detected, but five continued on BPs for 2 years. Localized reaction deterioration was more common in patients who continued than discontinued BPs (100 vs. 29%; p = 0.027). After 2 years, all patients had discontinued BPs and localized reaction did not deteriorate further in any patient. CONCLUSIONS: Once a localized reaction was detected, discontinuation of BPs and switching to vitamin D supplementation or teriparatide therapy effectively improved it. When the localized reaction was high, of the pointed type, and/or accompanied by prodromal pain, the risks of complete and incomplete AFF increased and consideration of prophylactic fixation for such patients was required.

The cumulative incidence of and risk factors for latent beaking in patients with autoimmune diseases taking long-term glucocorticoids and bisphosphonates.

SUMMARY: The incidence of beaking, which has been reported to precede atypical femoral fracture, was high and increased over 2 years in patients with autoimmune diseases who were taking bisphosphonates and glucocorticoids. Regular femoral X-rays are strongly recommended to screen for beaking, and bisphosphonate drug holidays should be considered. INTRODUCTION: Atypical femoral fractures (AFFs) have been recently recognized as complications associated with bisphosphonate (BP) use. AFFs are considered to be stress fractures; localized periosteal thickening of the lateral cortex is often present at the fracture site; this thickening is termed "beaking." Beaking has been reported to precede AFF. The aims of the present study were to evaluate the incidence of latent beaking in patients with autoimmune diseases taking BPs and glucocorticoids and to identify risk factors for beaking. METHODS: A total of 125 patients with autoimmune diseases who were taking BPs and glucocorticoids was included; 116 patients underwent X-rays and analysis of serum and urine bone metabolic markers annually for 2 years. Mean patient age was 54.5 years; there were 105 (90.5%) females and the mean duration of disease was 13.2 years. Focal lateral cortical thickening in femoral X-rays was defined as beaking. RESULTS: Beaking was detected in 15 femora of 10 patients (8.0%) at the time of recruitment. Over the 2-year observation period, the incidence of beaking increased to 21 femora of 12 patients (10.3%), and a complete AFF at the location of beaking occurred in one patient. Beaking was associated with a longer duration of BP treatment (6.1 ± 1.0 years vs. 5.0 ± 2.9 years, p = 0.01). Age 40-60 years, BP therapy ≥4 years, and diabetes mellitus were significantly associated with beaking. CONCLUSIONS: The incidence of beaking was high, and increased over 2 years, in patients with autoimmune diseases who were taking BPs and glucocorticoids. Regular femoral X-rays are strongly recommended to screen for beaking. Long-term BP/glucocorticoid use was a risk factor for beaking in patients with autoimmune diseases; BP drug holidays should be considered.

Improvement of depth dose distribution using multiple-field irradiation in boron neutron capture therapy.

It is important that improvements are made to depth dose distribution in boron neutron capture therapy, because the neutrons do not reach the innermost regions of the human body. Here, we evaluated the dose distribution obtained using multiple-field irradiation in simulation. From a dose volume histogram analysis, it was found that the mean and minimum tumor doses were increased using two-field irradiation, because of improved dose distribution for deeper-sited tumors.

The global economic crisis, household income and pre-adolescent overweight and underweight: a nationwide birth cohort study in Japan.

BACKGROUND: We hypothesized that children from lower income households and in households experiencing a negative income change in connection to the global economic crisis in 2008 would be at increased risk of adverse weight status during the subsequent years of economic downturn. METHODS: Data were obtained from a nationwide longitudinal survey comprising all children born during 2 weeks of 2001. For 16,403 boys and 15,206 girls, information about anthropometric measurements and household characteristics was collected from 2001 to 2011 on multiple occasions. Interactions between the crisis onset (September 2008) and household income group, as well as the crisis onset and a >30% negative income change in connection to the crisis, were assessed with respect to risk of childhood over- and underweight. RESULTS: Adjusted for household and parental characteristics, boys and girls in the lower household income quartiles had a larger increase in risk of overweight after the crisis onset relative to their peers in the highest income group. (Odds ratio (95% confidence interval) for interaction term in boys=1.23 (1.02-1.24); girls=1.35 (1.23-1.49) comparing the lowest with the highest income group.) Among girls, an interaction between the crisis onset and a >30% negative change in household income with respect to risk of overweight was observed (odds ratio for interaction term=1.23 (1.09-1.38)). Girls from the highest income group had an increased risk of underweight after the crisis onset compared with girls from the lowest income group. CONCLUSIONS: Boys and girls from lower household income groups and girls from households experiencing a negative income change in connection to the global economic crisis in 2008, may be at increased risk of overweight. Vulnerability to economic uncertainty could increase risk of overweight in preadolescence.

Influence of exercise training with thigh compression on heat-loss responses.

We investigated the effect of thigh compression, which accelerates activation of central command and muscle metabo- and mechanoreceptors, on the adaptation of sweating and cutaneous vascular responses during exercise heat acclimation. Nine non-heat-acclimated male subjects were acclimated to heat (32 °C and 50% RH) while cycling [50% of maximum oxygen uptake ( V ˙ O 2 m a x )] 60 min/day for 7 days (control group). The experimental group (n = 9) conducted the same training while the proximal thighs were compressed by a cuff at 60 mmHg. V ˙ O 2 m a x , acetylcholine-induced forearm sweating rate (iontophoresis), and mean sweating and cutaneous vascular responses on the forehead, chest, and forearm (SRmean and CVCmean ) during passive heating were evaluated before and after training. Training significantly increased V ˙ O 2 m a x while did not affect acetylcholine-induced sweating rates in either group. Training significantly decreased Tb thresholds for SRmean and CVCmean during passive heating without the alternations of sensitivities in both groups. Although SRmean during passive heating at a given ΔTb was not improved in either group, CVCmean was significantly (P < 0.05) attenuated after exercise training only in experimental group. Our results indicate that thigh cuff compression during exercise heat acclimation does not influence adaptation of the sweating response but attenuate cutaneous vasodilation.

The Piccolo Intronic Single Nucleotide Polymorphism rs13438494 Regulates Dopamine and Serotonin Uptake and Shows Associations with Dependence-Like Behavior in Genomic Association Study.

Piccolo (PCLO) inhibits methamphetamine-induced neuropharmacological effects via modulation of dopamine (DA) uptake and regulation of the transport of synaptic vesicles in neuronal cells. Clinical studies have recently suggested that the single nucleotide polymorphism (SNP) rs13438494 in the intron 24 of the PCLO gene is associated with psychiatric disorder, in the meta-analysis of GWAS. Therefore, in this study, we attempted to evaluate the possible role of the PCLO SNP in the mechanisms of uptake of monoamines. To characterize rs13438494 in the PCLO gene, we constructed plasmids carrying either the C or A allele of the SNP and transiently transfected them into SH-SY5Y cells to analyze genetic effects on the splicing of PCLO mRNA. The C and A allele constructs produced different composition of the transcripts, indicating that the intronic SNP does affect the splicing pattern. We also transfected DA and serotonin (5-hydroxytryptamine; 5- HT) transporters into cells and analyzed their uptakes to elucidate the association to psychiatric disorders. In the cells transfected with the C allele, both the DA and 5-HT uptake were enhanced compared to the A allele. We also conducted a clinical study, in order to clarify the genetic associations. PCLO rs13438494 exhibits a relationship with the symptoms of drug dependence or related parameters, such as the age of first exposure to methamphetamine, eating disorders, tobacco dependence and fentanyl requirement. Our findings suggest that rs13438494 is associated with drug abuse and contributes to the pathogenesis of psychiatric disorders via modulation of neurotransmitter turnover.

Quantitative characterization of hydration state and destructuring effect of monosaccharides and disaccharides on water hydrogen bond network.

Terahertz time-domain attenuated total reflection measurements of monosaccharide (glucose and fructose) and disaccharide (sucrose and trehalose) solutions from 0.146 M to 1.462 M were performed to evaluate (1) the hydration state and (2) the destructuring effect of saccharide solutes on the hydrogen bond (HB) network. Firstly, the extent of hydration water was determined by the decreased amount of bulk water with picosecond relaxation time that was replaced by that with much longer orientational relaxation time. As a result, we found glucose and trehalose exhibits stronger hydration capacity than fructose and sucrose, respectively, despite of the same number of the hydroxyl groups. For each saccharide, the hydration number tended to decrease with solute concentration. Secondly, the destructuring effect of these saccharide solutes on the HB network of the surrounding bulk water was discussed from the perspective of the fraction of non-hydrogen-bonded (NHB) water isolated from the HB network. We found the fraction of NHB water molecules that are not engaged in the HB network monotonously increased with saccharide concentration, indicating saccharide solutes promote the disruption of the water HB network. However, no noticeable differences were confirmed in the fraction of NHB water between glucose and fructose or between sucrose and trehalose. In contrast to hydration number, the number of NHB water produced by a single saccharide solute was less dependent on solute concentration, and three monosaccharide/disaccharide solutes were found to produce one/two NHB water molecules.

Hydration and hydrogen bond network of water around hydrophobic surface investigated by terahertz spectroscopy.

Water conformation around hydrophobic side chains of four amino acids (glycine, L-alanine, L-aminobutyric acid, and L-norvaline) was investigated via changes in complex dielectric constant in the terahertz (THz) region. Each of these amino acids has the same hydrophilic backbone, with successive additions of hydrophobic straight methylene groups (-CH2-) to the side chain. Changes in the degree of hydration (number of dynamically retarded water molecules relative to bulk water) and the structural conformation of the water hydrogen bond (HB) network related to the number of methylene groups were quantitatively measured. Since dielectric responses in the THz region represent water relaxations and water HB vibrations at a sub-picosecond and picosecond timescale, these measurements characterized the water relaxations and HB vibrations perturbed by the methylene apolar groups. We found each successive straight -CH2- group on the side chain restrained approximately two hydrophobic hydration water molecules. Additionally, the number of non-hydrogen-bonded (NHB) water molecules increased slightly around these hydrophobic side chains. The latter result seems to contradict the iceberg model proposed by Frank and Evans, where water molecules are said to be more ordered around apolar surfaces. Furthermore, we compared the water-hydrophilic interactions of the hydrophilic amino acid backbone with those with the water-hydrophobic interactions around the side chains. As the hydrophobicity of the side chain increased, the ordering of the surrounding water HB network was altered from that surrounding the hydrophilic amino acid backbone, thereby diminishing the fraction of NHB water and ordering the surrounding tetrahedral water HB network.