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Promyelocytic leukemia-nuclear bodies (PML-NBs) are dot-like protein assemblies and implicated in the pathogenesis of leukemia and viral infection. PML is the scaffold protein of PML-NB and its client proteins such as SUMO, DAXX, and Sp100 reside in PML-NBs. It is known that a short exposure to trivalent arsenic (As3+) induces the solubility change and the subsequent SUMOylation of PML, and the SUMO interacting motif (SIM) is not necessary for these biochemical changes. However, it has not been well studied how As3+initiates or enhances the association of SUMO with PML and the other PML-NB client proteins. Here, we report that As3+enhanced non-covalent association of PML with SUMO via the SUMO-SIM interaction which is dispensable for the solubility change and SUMOylation of PML. We also report that the As3+-induced solubility change of PML was not affected by ML792, a SUMO E1 enzyme inhibitor, even though the nuclear localization of SUMO2/3 and protein SUMOylation were halted by ML792. As3+did not change the solubility of DAXX and SUMOylation enzymes such as SAE1, UBA2, and UBC9. In contrast, As3+induced SUMOylation of Sp100 with a concomitant loss of its solubility like PML in human leukemia cell lines. Our current results indicate that both covalent and non-covalent associations of SUMO with PML are increased in As3+-exposed cells, and Sp100 may play a role in the maintenance of PML-NBs.

Asparagus is a popular vegetable rich in healthy functional components. However, the process of its production leaves ferns from aboveground parts and roots from underground parts as unusable parts, and this is an issue to be resolved. In our previous studies, large amounts of rutin were noted in the cladophylls and storage roots (brown and epidermis), and the protodioscin content was high in buds, in the soil-covered section of spears, and in rhizomes. This study was conducted to examine the distribution of growth-inhibitory activity and mineral contents in different parts of asparagus. Correlations, including representative functional components (rutin and protodioscin), were examined. The results suggest there are differences in growth-inhibitory activity of different parts of asparagus. The growth-inhibitory activity was strong in the buds, rhizome, and absorptive and storage roots, and weak in the cladophylls and lateral branches. The percent N content of the aboveground part of asparagus was high compared with that in the aboveground part of other crops. Although the percent K content was similar to the mean of the aboveground part of other crops, it was higher than that in general green manure, suggesting the residual stems and leaves of the aboveground part of asparagus are effective green manure. In the aboveground part of asparagus, the rutin content and percent N and K content were higher, whereas growth-inhibitory activity tended to be low, suggesting that when no disease developed in the aboveground part, it can be used as an organic substance.

Updating the value of sensory cues through experiences is a critical element of adaptive behavior. Although dopaminergic neurons (DANs) achieve this by driving associative learning, whether they contribute to assessment of sensory values outside the context of association remains largely unexplored. Here we show in Drosophila that DANs in the mushroom body encode the innate value of odors and constantly update the current value by inducing plasticity during olfactory navigation. Simulation of neuronal activity in a network based on the connectome data reproduced the characteristics of DAN responses, proposing a concrete circuit mechanism for computation. Notably, odors alone induced value- and dopamine-dependent changes in the activity of mushroom body output neurons, which store the current value of odors, as well as the behavior of flies steering in a virtual environment. Thus, the DAN circuit known for discrete, associative learning also continuously updates odor values during navigation in a non-associative manner.

Difficulty in cessation of drinking, smoking, or gambling has been widely recognized. Conventional theories proposed relative dominance of habitual over goal‐directed control, but human studies have not convincingly supported them. Referring to the recently suggested “successor representation (SR)” of states that enables partially goal‐directed control, we propose a dopamine‐related mechanism that makes resistance to habitual reward‐obtaining particularly difficult. We considered that long‐standing behavior towards a certain reward without resisting temptation can (but not always) lead to a formation of rigid dimension‐reduced SR based on the goal state, which cannot be updated. Then, in our model assuming such rigid reduced SR, whereas no reward prediction error (RPE) is generated at the goal while no resistance is made, a sustained large positive RPE is generated upon goal reaching once the person starts resisting temptation. Such sustained RPE is somewhat similar to the hypothesized sustained fictitious RPE caused by drug‐induced dopamine. In contrast, if rigid reduced SR is not formed and states are represented individually as in simple reinforcement learning models, no sustained RPE is generated at the goal. Formation of rigid reduced SR also attenuates the resistance‐dependent decrease in the value of the cue for behavior, makes subsequent introduction of punishment after the goal ineffective, and potentially enhances the propensity of nonresistance through the influence of RPEs via the spiral striatum‐midbrain circuit. These results suggest that formation of rigid reduced SR makes cessation of habitual reward‐obtaining particularly difficult and can thus be a mechanism for addiction, common to substance and nonsubstance reward., Long‐standing behavior towards a certain reward can lead to a formation of rigid reduced successor representation (SR) of states based on the future occupancy of the rewarded goal state. If it is formed, whereas no reward prediction error (RPE) is generated at the goal while the person does not resist temptation, once the person starts resistance, a sustained large RPE is generated at the goal, somewhat similarly to the hypothesized sustained fictitious RPE caused by drug‐induced dopamine.

Hypertension is the most considerable but treatable risk factor for cardiovascular disease. Although physicians prescribe multiple antihypertensive drugs and promote lifestyle modifications, the real‐world blood pressure (BP) control rate remains poor. To improve BP target achievement, we developed a novel digital therapeutic—the HERB software system —to manage hypertension. Here, we performed a randomized pilot study to assess the safety and efficacy of the HERB system for hypertension. We recruited 146 patients with essential hypertension from March 2018 to March 2019. We allocated eligible patients to the intervention group (HERB system + standard lifestyle modification) or control group (standard lifestyle modification alone). The primary outcome was the mean change from baseline to 24 weeks in 24‐hour systolic BP (SBP) measured by ambulatory blood pressure monitoring (ABPM). The baseline characteristics in each group were well balanced; the mean age was approx. 57 years, and 67% were male. In the primary end point at 24 weeks, HERB intervention did not lower the mean change of 24‐hour SBP by ABPM compared with the controls (adjusted difference: −0.66 mmHg; p = .78). In an exploratory analysis focusing on antihypertensive drug‐naïve patients aged

Dopamine signals ramping towards reward timings have become widely reported, but their functions remain elusive. Through modeling analyses and experiments in mice, a recent study by Mikhael, Kim et al. shows that such signals represent reward prediction errors used for accurate value learning in conditions with uncertainty about upcoming state and its resolution by sensory feedback.

Promyelocytic leukemia (PML) nuclear bodies (PML-NBs), a class of membrane-less cellular organelles, participate in various biological activities. PML-NBs are known as the core-shell-type nuclear body, harboring ‘client’ proteins in their core. Although multiple membrane-less organelles work in the oocyte nucleus, PML-NBs have been predicted to be absent from oocytes. Here, we show that some well-known PML clients (but not endogenous PML) co-localized with small ubiquitin-related modifier (SUMO) protein in the nucleolus and peri-centromeric heterochromatin of maturing oocytes. In oocytes devoid of PML-NBs, endogenous PML protein localized in the vicinity of chromatin. During and after meiotic resumption, PML co-localized with SUMO gathering around chromosomes. To examine the benefit of the PML-NB-free intranuclear milieu in oocytes, we deliberately assembled PML-NBs by microinjecting human PML-encoding plasmids into oocytes. Under conditions of limited SUMO availability, assembled PML-NBs tended to cluster. Upon proteotoxic stress, SUMO delocalized from peri-centromeric heterochromatin and co-localized with SC35 (a marker of nuclear speckles)-positive large compartments, which was disturbed by pre-assembled PML-NBs. These observations suggest that the PML-NB-free intranuclear environment helps reserve SUMO for emergent responses by redirecting the flux of SUMO otherwise needed to maintain PML-NB dynamics.

Promyelocytic leukemia (PML) nuclear bodies (PML-NBs), a class of membrane-less organelles in cells, are involved in multiple biological activities and are present throughout cells of adult organisms. Although the oocyte nucleus is an active region for the flux of multiple non-membranous organelles, PML-NBs have been predicted to be absent from oocytes. Here, we show that the deliberate assembly of PML-NBs during oocyte growth preferentially sequestered Small Ubiquitin-related Modifier (SUMO) protein from the nucleoplasm. SUMO not only was involved in the regulation of oocyte nuclear maturation but also was committed to the response, mediated by liquid droplet formation, to multiple stressors including nucleolar stress and proteotoxic stresses. Exogenous assembly of PML-NBs in the nucleus of oocytes affected the efficiency of the response of SUMO. These observations suggest that the PML-NB-free intranuclear milieu ensures that a reserve of SUMO remains available for emergent responses in oocyte development. This work demonstrated a benefit of the PML-NB-free intranuclear milieu, namely the ability to redirect the flux of SUMO otherwise needed to control PML-NB dynamics.

Asparagus is a popular vegetable rich in healthy functional components. Asparagus spears are known to contain a large amount of rutin, which has been found to possess anti-inflammatory, antitumor, and antibacterial/viral properties, and protodioscin, which is an antitumor substance and present in the bottom parts (8 cm from the cut end). However, the process of its production leaves fern in the aboveground parts and roots in the underground parts as significant amounts of nonusable parts, and this issue should be solved. This study was conducted to examine the distributions of rutin and protodioscin, representative functional components in different parts of asparagus. The results suggested that large amounts of rutin were noted in the cladophylls and storage roots (brown and epidermis), and the protodioscin content was high in the buds, the soil-covered section of the spear, and the rhizome. A significant amount of rutin was detected in the aboveground parts, which is consistent with the results of previous studies, but it was also found in the storage roots. The largest amount of protodioscin was found in the buds, as well as in young fruits and seeds of the aboveground parts. Injury by continuously cropping asparagus may be associated with high rutin content in the storage roots of asparagus.

The pregnane X receptor (PXR) is the key regulator of our defense mechanism against foreign substances such as drugs, dietary nutrients, or environmental pollutants. Because of increased health consciousness, the use of dietary supplements has gradually increased, and most of them can activate PXR. Therefore, an analysis of the interaction between drugs and nutrients is important because altered levels of drug-metabolizing enzymes or transporters can remarkably affect the efficiency of a co-administered drug. In the present study, we analyzed the effect of vitamin K-mediated PXR activation on drug metabolism-related gene expression in intestine-derived LS180 cells via gene expression studies and western blotting analyses. We demonstrated that menaquinone 4 (MK-4), along with other vitamin Ks, including vitamin K1, has the potential to induce MDR1 and CYP3A4 gene expression. We showed that PXR knockdown reversed MK-4-mediated stimulation of these genes, indicating the involvement of PXR in this effect. In addition, we showed that the expression of MDR1 and CYP3A4 genes increased synergistically after 24 h of rifampicin and MK-4 co-treatment. Our study thus elucidates the importance of drug–nutrient interaction mediated via PXR.

Neurosarcoidosis is a rare disease and is often difficult to diagnose. Herein, we report a case of neurosarcoidosis in a patient with a history of Ewing’s sarcoma of the brain. He presented with fever of unknown origin, and a pathological diagnosis was obtained via biopsy of a normal-sized inguinal lymph node with fluorodeoxyglucose (FDG) accumulation on positron emission tomography/computed tomography (PET/CT). The condition could not have been diagnosed without FDG-PET/CT.

Background: We previously showed 8-weeks of fish oil supplementation attenuated muscle damage. However, the effect of a shorter period of fish oil supplementation is unclear. The present study investigated the effect of fish oil, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), for 4 weeks on muscular damage caused by eccentric contractions (ECCs) of the elbow flexors.Methods: Twenty-two untrained men were recruited in this double-blind, placebo-controlled, parallel design study and the subjects were randomly assigned to the EPA and DHA group (EPA and DHA, n = 11) and placebo group (PL, n = 11). They consumed either EPA 600 mg and DHA 260 mg per day or placebo supplement for 4 weeks prior to exercise. Subjects performed 60 ECCs at 100% maximal voluntary contraction (MVC) using a dumbbell. Changes in MVC torque, range of motion (ROM), upper arm circumference, muscle soreness, echo intensity, muscle thickness, serum creatine kinase (CK), and interleukin-6 (IL-6) were assessed before exercise; immediately after exercise; and 1, 2, 3, and 5 days after exercise.Results: ROM was significantly higher in the EPA and DHA group than in the PL group immediately after performing ECCs (p < 0.05). No differences between groups were observed in terms of MVC torque, upper arm circumference, muscle soreness, echo intensity, and thickness. A significant difference was observed in serum CK 3 days after ECCs (p < 0.05).Conclusion: We concluded that shorter period EPA and DHA supplementation benefits joint flexibility and protection of muscle fiber following ECCs.

In order to analyze the dynamic behavior of neural systems such as oscillations or rhythms, an approach based on the dynamical systems theory may be useful. In the first part of this article, we present an elementary introduction to that approach based on the materials for an author's lecture. In the second part, we introduce our own study related to dopamine and reinforcement learning using that approach, in which we assumed the decay of learned values and propose a possible mechanism of the effects of dopamine depletion on motivation.

Difficulty in cessation of drinking, smoking, or gambling has been widely recognized. Conventional theories proposed relative dominance of habitual over goal-directed control, but human studies have not convincingly supported them. Referring to the recently suggested “successor representation” of states that enables partially goal-directed control, we propose a dopamine-related mechanism potentially underlying the difficulty in resisting habitual reward-seeking, common to substance and non-substance reward. Consider that a person has long been taking a series of actions leading to a certain reward without resisting temptation. Given the suggestions of the successor representation and the dimension reduction in the brain, we assumed that the person has acquired a dimension-reduced successor representation of states based on the goal state under the established non-resistant policy. Then, we show that if the person changes the policy to resist temptation, a large positive reward prediction error (RPE) becomes generated upon eventually reaching the goal, and it sustains given that the acquired state representation is so rigid that it does not change. Inspired by the anatomically suggested spiral striatum-midbrain circuit and the theoretically proposed spiraling accumulation of RPE bias in addiction, we further simulated the influence of RPEs generated in the goal-based representation system on another system representing individual actions. We then found that such an influence could potentially enhance the propensity of non-resistant choice. These results suggest that the inaccurate value estimation in the reduced successor representation system and its influence through the spiral striatum-midbrain circuit might contribute to the difficulty in cessation of habitual reward-seeking.

Background After the accident at the Fukushima Daiichi Nuclear Power Plant, radioactive contaminants were released over a widespread area. Monitoring the biological effects of radiation exposure in animals in the ex-evacuation zone should be continued to understand the health effects of radiation exposure in humans. The present study aimed to clarify the effects of radiation by investigating whether there is any alteration in the morphology and gene expressions of immune molecules in the intestine of pigs and inobuta (wild boar and domestic pig hybrid) in the ex-evacuation zone in 2012. Gene expression analysis was performed in small intestine samples from pigs, which were collected from January to February 2012, in the ex-evacuation zone. Pigs lived freely in this zone, and their small intestine was considered to be affected by the dietary intake of radioactive contaminants. Results Several genes were selected by microarray analysis for further investigation using real-time polymerase chain reaction. IFN-γ, which is an important inflammatory cytokine, and TLR3, which is a pattern recognize receptor for innate immune system genes, were highly elevated in these pigs. The expressions of the genes of these proteins were associated with the radiation level in the muscles. We also examined the alteration of gene expressions in wild boars 5 years after the disaster. The expression of IFN-γ and TLR3 remained high, and that of Cyclin G1, which is important in the cell cycle, was elevated. Conclusions We demonstrated that some changes in gene expression occurred in the small intestine of animals in the ex-evacuation zone after radiation. It is difficult to conclude that these alterations are caused by only artificial radionuclides from the Fukushima Daiichi Nuclear Power Plant. However, the animals in the ex-evacuation zone might have experienced some changes owing to radioactive materials, including contaminated soil, small animals, and insects. We need to continue monitoring the effects of long-term radiation exposure in living things.

Inorganic arsenicals are well-known carcinogens, whereas arsenite (iAsIII) compounds are now recognized as potent therapeutic agents for several leukemias, and arsenic trioxide has been used for the treatment of recurrent acute promyelocytic leukemia (APL). However, recent clinical trials revealed that arsenite is not always effective for non-APL malignancies. Another arsenical, S-dimethylarsino-glutathione ([DMAIII(GS)]), which is a putative metabolic intermediate in the hepatic metabolism of iAsIII, shows promise for treating several types of lymphoma. However, the metabolism of [DMAIII(GS)] has not been well investigated, probably because [DMAIII(GS)] is not stable in biological fluids where the concentration of glutathione is low. In the present study, we injected [DMAIII(GS)] intravenously into mice and compared the tissue distribution and metabolic dynamics of [DMAIII(GS)] with those of sodium arsenite (NaAsO2). We found a unique organ preference for the distribution of [DMAIII(GS)] to the lung and brain in comparison to NaAsO2. Furthermore, [DMAIII(GS)] appeared to bind to serum albumin by exchanging its glutathione moiety quickly after administration, providing novel insights into the longer retention of [DMAIII(GS)] in plasma.

Background Smoking cessation helps extend a healthy life span and reduces medical expenses. However, the standard 12-week smoking cessation program in Japan has several notable problems. First, only 30% of participants complete this program. Second, participants may choose not to participate unless they have a strong motivation to quit smoking, such as health problems. Third, the program does not provide enough support during the period between clinical visits and after 12 weeks. Objective This study examined the efficacy of the 24-week ascure program to address the problems of accessibility and continuous support. The program combines online mentoring, over-the-counter pharmacotherapy, and a smartphone app. Methods Using a retrospective study design, we investigated data for 177 adult smokers who were enrolled in the ascure smoking cessation program between August 2017 and August 2018. The primary outcomes were continuous abstinence rates (CARs) during weeks 9-12 and weeks 21-24. To confirm smoking status, we performed salivary cotinine testing at weeks 12 and 24. We also evaluated the program adherence rate. Finally, we performed exploratory analysis to determine the factors associated with continuous abstinence at weeks 21-24 to provide insights for assisting with long-term continuous abstinence. Results The CARs of all participants for weeks 9-12 and weeks 21-24 were 48.6% (95% CI 41.2-56.0) and 47.5% (95% CI 40.0-54.8), respectively. Program adherence rates were relatively high throughout (72% at week 12 and 60% at week 24). In the analysis of the factors related to the CAR at weeks 21-24, the number of entries in the app’s digital diary and number of educational videos watched during the first 12 weeks were significant factors. Conclusions The ascure program achieved favorable CARs, and participants showed high adherence. Proactive usage of the smartphone app may help contribute to smoking cessation success in the long-term.

BACKGROUND Smoking cessation helps extend a healthy life span and reduces medical expenses. However, the standard 12-week smoking cessation program in Japan has several notable problems. First, only 30% of participants complete this program. Second, participants may choose not to participate unless they have a strong motivation to quit smoking, such as health problems. Third, the program does not provide enough support during the period between clinical visits and after 12 weeks. OBJECTIVE This study examined the efficacy of the 24-week ascure program to address the problems of accessibility and continuous support. The program combines online mentoring, over-the-counter pharmacotherapy, and a smartphone app. METHODS Using a retrospective study design, we investigated data for 177 adult smokers who were enrolled in the ascure smoking cessation program between August 2017 and August 2018. The primary outcomes were continuous abstinence rates (CARs) during weeks 9-12 and weeks 21-24. To confirm smoking status, we performed salivary cotinine testing at weeks 12 and 24. We also evaluated the program adherence rate. Finally, we performed exploratory analysis to determine the factors associated with continuous abstinence at weeks 21-24 to provide insights for assisting with long-term continuous abstinence. RESULTS The CARs of all participants for weeks 9-12 and weeks 21-24 were 48.6% (95% CI 41.2-56.0) and 47.5% (95% CI 40.0-54.8), respectively. Program adherence rates were relatively high throughout (72% at week 12 and 60% at week 24). In the analysis of the factors related to the CAR at weeks 21-24, the number of entries in the app’s digital diary and number of educational videos watched during the first 12 weeks were significant factors. CONCLUSIONS The ascure program achieved favorable CARs, and participants showed high adherence. Proactive usage of the smartphone app may help contribute to smoking cessation success in the long-term.

It is extremely important to understand the mechanisms underlying the effect of radiation on animals including humans. Radiation potentially induces alterations of the immune system, which may cause serious diseases. The Fukushima Daiichi Nuclear Power Plant (FNPP) accident occurred in 2011, in which radioactive contaminants were released over a wide area. After farmers evacuated from the ex-evacuation zone set within 20-km radius from FNPP, pigs were left behind and unleashed in the zone (abandoned pigs). Their small intestine was considered to have been affected by the dietary intake of radioactive contaminants. The present study thus aimed to clarify the effect of radiation by investigating whether there is any alteration in the expression of genes encoding immune-related molecules and in morphology of the intestine of abandoned pigs. Microarray analysis revealed changes of the expression of various genes involved in inflammation and oxidative stress including IFN-γ, which is an important inflammatory cytokine, was highly elevated in abandoned pigs, and its expression level was correlated with radioactive cesium concentration in skeletal muscle. On the other hand, there were no morphological changes in the intestine. At the sampling stage of this study, only 1 year passed since the FNPP accident occurred, it would be premature to conclude that the identified alteration of gene expression was caused only by artificial radionuclides attributed to the FNPP accident. We need to continue monitoring the effect of long-term radiation exposure in living organisms in the ex-evacuation zone.

Promyelocytic leukemia (PML) and a suite of other proteins form nuclear bodies (NBs) where SUMOylation of PML and tumor suppression events occur in response to arsenite (As3+) treatment. Soluble PML is rapidly modified to the insoluble form in response to As3+, yet the relationship between the solubility change and nuclear localization of PML and PML-nuclear body (PML-NB) proteins remained elusive. We have investigated differences in the solubility change of well-known PML-NB proteins such as death-associated protein 6 (DAXX), SUMO, and PML in genetically engineered HEK293, and Jurkat and HL60 cells. The solubility of PML and SUMO2/3 monomers in RIPA solution decreased in 2 h in response to As3+. Live image analysis of GFP-PML revealed that extranuclear PML was insoluble in RIPA irrespective of the As3+-treatment and PML in PML-NBs, which was soluble in the untreated cells, was converted to insoluble forms by As3+. The solubility of DAXX was not changed by As3+, even though PML and DAXX co-localized completely in the subcellular compartments. Murine double mutant 2 (MDM2), which is known to interacts with intranuclear PML, did not affect the As3+-induced solubility change of PML. These results indicate that As3+ selectively reorganizes PML and SUMO2/3 monomers into insoluble forms in PML-NBs, and then PML SUMOylation proceeds.

Promyelocytic leukemia (PML), which is a tumor suppressor protein that nevertheless plays an important role in the maintenance of leukemia initiating cells, is known to be biochemically modified by As{sup 3+}. We recently developed a simple method to evaluate the modification of PML by As{sup 3+} resulting in a change in solubility and the covalent binding of small ubiquitin-like modifier (SUMO). Here we semi-quantitatively investigated the SUMOylation of PML using HEK293 cells which were stably transfected with PML-VI (HEK-PML). Western blot analyses indicated that PML became insoluble in cold RadioImmunoPrecipitation Assay (RIPA) lysis buffer and was SUMOylated by both SUMO2/3 and SUMO1 by As{sup 3+}. Surprisingly SUMO1 monomers were completely utilized for the SUMOylation of PML. Antimony (Sb{sup 3+}) but not bismuth (Bi{sup 3+}), Cu{sup 2+}, or Cd{sup 2+} biochemically modified PML similarly. SUMOylated PML decreased after removal of As{sup 3+} from the culture medium. However, unSUMOylated PML was still recovered in the RIPA-insoluble fraction, suggesting that SUMOylation is not requisite for changing the RIPA-soluble PML into the RIPA-insoluble form. Immunofluorescence staining of As{sup 3+}-exposed cells indicated that SUMO2/3 was co-localized with PML in the nuclear bodies. However, some PML protein was present in peri-nuclear regions without SUMO2/3. Functional Reallymore » Interesting New Gene (RING)-deleted mutant PML neither formed PML nuclear bodies nor was biochemically modified by As{sup 3+}. Conjugation with intracellular glutathione may explain the accessibility of As{sup 3+} and Sb{sup 3+} to PML in the nuclear region evading chelation and entrapping by cytoplasmic proteins such as metallothioneins. - Highlights: • As{sup 3+} is a carcinogen and also a therapeutic agent for leukemia. • PML becomes insoluble in RIPA and SUMOylated by As{sup 3+}. • Sb{sup 3+} modifies PML similar to As{sup 3+}. • Functional RING motif is necessary for As{sup 3+}-induced PML modification.« less

There are several mechanisms for antibiotic-resistant Pseudomonas aeruginosa. The purpose of this study is to investigate the association between the expression of efflux pump-coding genes and antibiotic resistance in P. aeruginosa causing urinary tract infections (UTIs). We extracted the RNA from 105 clinical strains of P. aeruginosa isolated from UTI patients with full data on antibiotic MICs and assayed real-time quantitative reverse-transcription PCR. We investigated the gene expressions of four resistance nodulation cell division-type multi-drug efflux pump systems (MexAB-OprM, MexCD-OprJ, MexEF-OprN and MexXY(-OprA)) and the correlation of the MICs of nine antibiotics, risk factors and antibiotic resistance-related genes with expressions of mexB, mexC, mexE and mexY. Multivariate statistical data demonstrated a significant relationship between increased expression of mexB or mexC and complicated UTI (Odds ratio=8.03, P

Highlights • Mutation of Cys227 in mouse AQP11 has been reported to cause kidney injury. • The importance of Cys227 for the molecular function of AQP11 has yet to be elucidated. • We examined the molecular function of Cys227 AQP11 in transfected mammalian cells. • The mutation at Cys227 caused an increase in the water permeability of AQP11. • Cys227 is considered to be crucial for the proper molecular function of AQP11., Aquaporin-11 (AQP11) is the latest member of the mammalian water channel protein family to be described. Recent in vivo studies have shown that mutation at Cys227 causes renal failure. However the importance of Cys227 for the molecular function of AQP11 is largely unknown. In this study, we examined the subcellular localization, water permeability, and multimerization of AQP11 with a mutation at Cys227. Interestingly, cells expressing the mutants had significantly higher osmotic water permeability. In contrast, the mutation lowered the cell surface expression and multimerization levels. Our observations suggest that Cys227 is crucial for the proper molecular function of AQP11.

Ribosomal DNA (rDNA) sequences are widely used for phylogenetic and bacterial identification. However, rDNA of different species often reveals similar or identical same sequences. This study employed the bacterial stable small RNA (ssrA) gene encoding transfer-messenger RNA (tmRNA) as a tool for identification of Staphylococcus aureus, Enterococcus spp, Pseudomonas spp and Enterobacteriaceae from clinical isolates as representative groups using PCR and species specific primers. The method correctly identified 11 standard strains and 99 clinical isolates. Quantitative PCR revealed a limit of detection of 10(-5) µg of DNA for S. aureus and Enterococcus spp, and 10(-6) µg for Pseudomonas spp and Enterobacteriaceae. Further studies with a greater number of bacteria especially from clinical samples will need to be undertaken before this bacterial molecular marker can be applied in a clinical setting.

It has been suggested that the midbrain dopamine (DA) neurons, receiving inputs from the cortico-basal ganglia (CBG) circuits and the brainstem, compute reward prediction error (RPE), the difference between reward obtained or expected to be obtained and reward that had been expected to be obtained. These reward expectations are suggested to be stored in the CBG synapses and updated according to RPE through synaptic plasticity, which is induced by released DA. These together constitute the 'DA=RPE' hypothesis, which describes the mutual interaction between DA and the CBG circuits and serves as the primary working hypothesis in studying reward learning and value-based decision making. However, recent work has revealed a new type of DA signal that appears not to represent RPE. Specifically, it has been found in a reward-associated maze task that striatal DA concentration primarily shows a gradual increase towards the goal. We explored whether such ramping DA could be explained by extending the 'DA=RPE' hypothesis by taking into account biological properties of the CBG circuits. In particular, we examined effects of possible time-dependent decay of DA-dependent plastic changes of synaptic strengths by incorporating decay of learned values into the RPE-based reinforcement learning model and simulating reward learning tasks. We then found that incorporation of such a decay dramatically changes the model's behavior, causing gradual ramping of RPE. Moreover, we further incorporated magnitude-dependence of the rate of decay, which could potentially be in accord with some past observations, and found that near-sigmoidal ramping of RPE, resembling the observed DA ramping, could then occur. Given that synaptic decay can be useful for flexibly reversing and updating the learned reward associations, especially in case the baseline DA is low and encoding of negative RPE by DA is limited, the observed DA ramping would be indicative of the operation of such flexible reward learning.

The purpose of this study was to examine the in vitro susceptibilities to antimicrobial agents and genetic diversity of 195 clinical strains of Salmonella spp., which were isolated and examined for the extended-spectrum β-lactamase (ESBL) blaCTX-M gene and the presence of gyrA, gyrB, parC, and parE genes mutations in Hyogo, Japan, from 2009 to 2012. Forty-three of the 195 strains were antimicrobial resistant. Two Salmonella enterica subsp. enterica strains, 1 serovar Schwarzengrund, and 1 serovar Enteritidis were identified as ESBL-producing strains possessing blaCTX-M-15 and blaCTX-M-2, respectively. Among 8 nalidixic acid-resistant strains, 7 had mutations in gyrA alone or in gyrA and parC. In conclusion, we identified CTX-M ESBL-producing Salmonella clinical strains with multidrug resistance. Further studies are needed to monitor these serious drug-resistant Salmonella strains in Japan.

A pvsB-vctA-irgA triple deletion mutant of Vibrio parahaemolyticus can utilize enterobactin under iron-limiting conditions by inducing a previously undescribed receptor, PeuA (VPA0150), in response to extracellular alkaline pH and enterobactin. In silico analyses revealed the existence of a two-component regulatory system operon, peuRS, immediately upstream of peuA, which constitutes an operon with the TonB2 system genes. Both the peuRS and peuA-tonB2 operons were found to be upregulated under iron-limiting conditions in a ferric uptake regulator (Fur)-dependent manner. The involvement of peuA and peuRS in enterobactin utilization was analyzed by complementation experiments using deletion mutants. Primer extension analysis indicated that, under iron-limiting conditions, the transcription of peuA was initiated from the +1 site at pH 7.0 and from both the +1 and +39 sites at pH 8.0 in the presence of enterobactin. The +39 transcript was absent from the peuRS deletion mutant. Secondary structure prediction of their 5′-untranslated regions suggested that translation initiation is blocked in the +1 transcript, but not in the +39 transcript. Consistent with this, in vitro translation analysis demonstrated that production of PeuA was determined only by the +39 transcript. These studies establish a novel gene regulation mechanism in which the two-component regulatory system PeuRS enhances expression of the alternative +39 transcript that possesses non-inhibitory structure, allowing the peuA expression to be regulated at the translation stage.

Fission yeast its3(+) encodes an essential phosphatidylinositol-4-phosphate 5-kinase (PI4P5K) that regulates cell integrity and cytokinesis. We performed a genetic screen to identify genes that function in PI4P5K-mediated signaling, and identified gyp10(+) encoding a Rab GTPase-activating protein (GAP), a negative regulator for Rab GTPase signaling. Its3 overproduction caused growth defects and abnormal cytoplasmic accumulation of the Its3 protein, which can be stained by calcofluor. Notably, Its3 overproducing cells displayed abnormal membranous structures, multilamella Golgi and fragmented vacuoles showed by Electron microscopy. Furthermore, the excess cytoplasmic Its3 structure partly colocalized with the fluorescence of FM4-64. Gyp10 rescued both growth defects and abnormal Its3 localization when it was over-expressed. Gyp10 functionally interacted with the Rab GTPases Ypt3 and Ryh1, both of which regulate Golgi membrane trafficking. Consistently, mutation or deletion of Ypt3 and Ryh1 suppressed phenotypes associated with Its3 overproduction. Importantly, the plasma membrane localization of Its3 was also affected by the impairment of the Ypt3/Ryh1 Rab membrane trafficking, thus suggesting that membrane trafficking events regulated by two Rab GTPases functionally interacts with PI4,5P2 signaling. These results suggest a mechanism whereby PI4P5K signaling/localization is affected by Golgi membrane trafficking, thus provide a functional link between the PI4,5P2 signaling and Rab-mediated trafficking.