Role of caspase-8 and/or -9 as biomarkers that can distinguish the potential to cause toxic- and immune related-adverse event, for the progress of acetaminophen-induced liver injury.

AIMS: Acetaminophen (APAP) overdose can cause acute liver failure. Although it is well known that APAP-induced liver injury (AILI) is caused by toxic mechanism, recently it is also reported to be immune related. However, the detail of the mechanism has been unclear. Therefore, elucidation of the pathophysiology is required. MAIN METHODS: In AILI model rats (800 mg/kg), the levels of AST, ALT and Caspase (C)-3/-8/-9 levels were measured. In in vitro study using human hepatocyte cells (FLC-4) and THP-1 cells, APAP (0.03-1.0 mM) were added to FLC-4 and the cell viability, C-9, cytochrome c, mitochondria membrane potential, and glutathione levels of FLC-4 and inflammasome activation of THP-1 were evaluated. KEY FINDINGS: In AILI model rats, the levels of AST and ALT were increased only at 12-24 h. C-3/-9 levels rose at 6-9 h, whereas C-8 level rose hours later, moreover, 24 h after; C-3/-8/-9 levels re-rose. In FLC-4 cells, cytochrome c was released from the mitochondria which is promoted by oxidative stress due to drug metabolism and C-9 was activated. Thus, AILI was caused mitochondrial damage by NAPQI as early reaction (first stage). In the next stage, inflammasomes of human antigen presenting cells, which released inflammatory cytokines were activated by damage-associated molecular patterns (DAMPs) released from damaged hepatocyte by APAP. SIGNIFICANCE: It is confirmed that AILI includes immune related mechanism. Thereby, in case of N-acetylcysteine refractory, additional administration of steroid hormones should be effective and recommended as a novel strategy for AILI with immune related adverse event (irAE).

Copulation induces expression of the immediate early gene Arc in mating-relevant brain regions of the male rat.

The medial amygdala (MeA), bed nucleus of the stria terminalis (BNST), and medial preoptic area (mPOA) are important for the regulation of male sexual behavior. Sexual experience facilitates sexual behaviors and influences activity in these regions. The goal of this study was to determine whether sexual experience or copulation induces plasticity in the MeA, BNST, or mPOA of male rats, as indicated by changes in levels of Arc, which is indicative of activity-dependent synaptic plasticity in the brain. To this end, sexually naïve or experienced males were placed in mating arenas either alone, with an inaccessible estrus female, or with an accessible estrus female. Arc protein levels were then quantified in these three regions using immunohistochemistry. As expected, sexual experience facilitated copulation, as evidenced by a reduction in latencies to mount, intromit, and ejaculate. Copulation also increased the number of Arc-positive cells in the MeA, anterior BNST, posterior BNST, and the posterior mPOA, but not in the central-rostral region of the mPOA. Surprisingly, prior sexual experience did not impact levels of Arc, suggesting that copulation-induced Arc occurs in both sexually naïve and experienced males.

Maternal and Child Health Handbook use for maternal and child care: a cluster randomized controlled study in rural Java, Indonesia.

BACKGROUND: Effectiveness of the Maternal and Child Health Handbook (MCHHB), a home-based booklet for pregnancy, delivery and postnatal/child health, was evaluated on care acquisition and home care in rural Java, a low service-coverage area. METHODS: We conducted a health centre-based randomized trial, with a 2-year follow-up. Intervention included (i) MCHHB provision at antenatal care visits; (ii) records and guides by health personnel on and with the MCHHB; and (iii) sensitization of care by volunteers using the MCHHB. RESULTS: The follow-up rate was 70.2% (183, intervention area; 271, control area). Respondents in the intervention area received consecutive MCH services including two doses of tetanus toxoid injections and antenatal care four times or more during pregnancy, professional assistance during child delivery and vitamin A supplements administration to their children, after adjustment for confounding variables and cluster effects (OR = 2.03, 95% CI: 1.19-3.47). In the intervention area, home care (continued breastfeeding; introducing complementary feeding; proper feeding order; varied foods feeding; self-feeding training; and care for cough), perceived support by husbands, and lower underweight rates and stunting rates among children were observed. CONCLUSION: MCHHB use promoted continuous care acquisition and care at home from pregnancy to early child-rearing stages in rural Java.

Estradiol in the Preoptic Area Regulates the Dopaminergic Response to Cocaine in the Nucleus Accumbens.

The sex-steroid hormone estradiol (E2) enhances the psychoactive effects of cocaine, as evidenced by clinical and preclinical studies. The medial preoptic area (mPOA), a region in the hypothalamus, is a primary neural locus for neuroendocrine integration, containing one of the richest concentrations of estrogen receptors in the CNS and also has a key role in the regulation of naturally rewarding behaviors. However, whether estradiol enhances the neurochemical response to cocaine by acting in the mPOA is still unclear. Using neurotoxic lesions and microdialysis, we examined whether the mPOA modulates cocaine-induced neurochemical activity in the nucleus accumbens. Tract tracing and immunohistochemical staining were used to determine whether projections from the mPOA to the ventral tegmental area (VTA) are sensitive to estrogen signaling. Finally, estradiol microinjections followed by microdialysis were used to determine whether estrogenic signaling in the mPOA modulates cocaine-induced changes of dopamine in the nucleus accumbens. Results showed that lesions of the mPOA or microinjections of estradiol directly into the mPOA increased cocaine-induced release of dopamine in the nucleus accumbens. Immunohistochemical analyses revealed that the mPOA modulates cocaine responsiveness via projections to both dopaminergic and GABAergic neurons in the VTA, and that these projections are sensitive to estrogenic stimulation. Taken together, these findings point to a novel estradiol-dependent pathway that modulates cocaine-induced neurochemical activity in the mesolimbic system.

Astrocytes in the medial preoptic area modulate ejaculation latency in an experience-dependent fashion.

While sexually experienced males copulate at a higher frequency than sexually inexperienced males, there is still a great deal of variability in their behavior. Within the medial preoptic area (mPOA) of the hypothalamus, glutamate modulates some of this variability. Glutamate levels, for example, increase during sexual activity, peaking with ejaculation and falling precipitously during the post-ejaculation interval. Whereas lower glutamate levels after ejaculation translates to longer post-ejaculatory intervals, administration of glutamate uptake inhibitors into the mPOA increases the number of ejaculations a male rat achieves over a mating bout, and reduces the latency to ejaculate once mating begins. Because astrocytes modulate the availability of neuronal glutamate, we hypothesized that differences in the number of GFAP-positive astrocytes in the mPOA may account for variability in sexual behavior. To this end, we examined whether the number of astrocytes in the mPOA related to ejaculation latency as well as to the duration of the post-ejaculatory interval (PEI) in sexually experienced and sexually inexperienced males. Results indicate that the number of astrocytes negatively correlated with latency to reach ejaculations in sexually inexperienced but not sexually experienced rats while the number of astrocytes and PEI were not related. Astrocyte numbers did not vary between inexperienced and experienced subjects indicating that astrocyte processes may differentially project to sex-relevant glutamatergic synapses or that glutamatergic innervation of the mPOA changes as a function of sexual experience.

Sexual experience influences mating-induced activity in nitric oxide synthase-containing neurons in the medial preoptic area.

Nitric oxide (NO) acts in the medial preoptic area (mPOA) of the hypothalamus to facilitate the expression of male sexual behavior and has also been widely implicated in mechanisms of experience, learning, and memory. Using immunohistochemistry for Fos, as a marker for neural activity, and nitric oxide synthase (NOS), the enzyme that catalyzes the production of nitric oxide (NO), we examined whether sexual activity and sexual experience influence Fos co-expression in NOS-containing neurons in the mPOA of male rats. Consistent with previous findings, results indicate that mating increased activity in the mPOA, and that sexual experience facilitated the expression of sexual behaviors, together with increased mating-induced Fos and NOS in the mPOA. Results also indicate that mating increased co-expression of Fos in NOS-containing neurons, and that this increase was highest in animals undergoing their first sexual encounter, indicating that initial sexual experience increases NO production in the mPOA of male rats.

Differences in forebrain androgen receptor expression in winners and losers of male anole aggressive interactions.

Size matched male green anoles (Anolis carolinensis) were paired in a neutral setting and allowed to engage in aggressive displays. Winners and losers were apparent in each pair within 90min, resulting in stable dominant/subordinate dyads. Androgen receptor (AR) expression was assessed at three time points after the initial pairing, 2h, 3 days, and 10 days in dominants, subordinates, and two groups of control males housed alone or with a female for an equal period of time. Expression was quantified in three forebrain areas that have been implicated in aggression and reproductive social behavior in this species, the preoptic area (POA), the anterior hypothalamus (AH), septal area (SEP), and ventromedial nucleus of the posterior division of the dorsal ventricular ridge (PDVRVM ). There were significant overall group differences in AR mRNA expression in the POA and AH that appeared to result from higher POA AR expression in dominant males compared to other groups, and generally lower AR expression in subordinate males. Pairwise comparison revealed that dominants' AR mRNA expression in the POA was significantly higher in the 2h and 3 day groups compared to that of subordinates, with a similar, but nonsignificant, difference in the 10 day group. Dominants had significantly higher AR mRNA expression in the AH compared to that of subordinates in the 2h group, but differences were not significant at later times. The results suggest that POA and AH sensitivity to androgens is increased in dominants compared to subordinates, and that the difference can be seen soon after the agonistic interaction establishing winners and losers.

Removal of toxic substances by a selective membrane plasma separator.

We devised a method of plasma exchange with dialysis (PED), in which selective plasma exchange (sPE) is performed using a selective membrane plasma separator (EC-2A) with an albumin-sieving coefficient of 0.3 while the dialysate flows outside the hollow fibers, and reported the usefulness of the system for treating acute liver failure. Thereafter, EC-4A with an albumin-sieving coefficient of 0.6 was developed, which was expected to be even more effective for removing protein-bound substances. In order to examine whether or not EC-4A might be applicable to blood purification therapy against drug poisoning, we compared the efficacies of sPE, PED, and direct hemoperfusion (DHP) using an activated carbon column for the removal of phenobarbital and lithium. Subjects undergoing the extracorporeal circulation study were assigned to the sPE group, PED group, or DHP group, and the changes in the blood concentrations of phenobarbital and lithium were measured over 180 min. A significant decrease of the phenobarbital concentration over time was seen in the PED group, as compared to that in the sPE group (P < 0.0001), while no significant difference in the concentration was observed between the PED and DHP groups. The PED group showed a significant decrease of the lithium concentration over time, as compared to the DHP group (P < 0.0001), while no significant difference in the concentration was observed between the PED and sPE groups. Thus, PED was as effective as DHP for removing phenobarbital and was as effective as sPE for removing lithium. These results suggest that PED therapy using EC-4A may be a feasible modality for the treatment of drug poisoning.

The role of home-based records in the establishment of a continuum of care for mothers, newborns, and children in Indonesia.

BACKGROUND: The provision of appropriate care along the continuum of maternal, newborn, and child health (MNCH) service delivery is a challenge in developing countries. To improve this, in the 1990s, Indonesia introduced the maternal and child health (MCH) handbook, as an integrated form of parallel home-based records. OBJECTIVE: This study aimed to identify the roles of home-based records both before and after childbirth, especially in provinces where the MCH handbook (MCHHB) was extensively promoted, by examining their association with MNCH service uptake. DESIGN: This was a cross-sectional study using nationally representative data sets, the Indonesia Demographic and Health Surveys (IDHSs) from 1997, 2002-2003, and 2007. The IDHS identifies respondents' ownership of home-based records before and after childbirth. Multivariate logistic regression was used to examine associations between record ownership and service utilisation in national data and data from two provinces, West Sumatra and North Sulawesi, where ownership of pre- and post-natal records served as a proxy for MCHHB ownership. RESULTS: Pre- and post-natal record ownership increased from 1997 to 2007. Provincial data from 2007 showed that handbook ownership was associated with having delivery assisted by trained personnel [adjusted odds ratio (aOR): 2.12, 95% confidence interval (CI): 1.05-4.25], receiving maternal care (aOR: 3.92, 95% CI: 2.35-6.52), completing 12 doses of child immunisation for seven diseases (aOR: 4.86, 95% CI: 2.37-9.95), and having immunisation before and after childbirth (aOR: 5.40, 95% CI: 2.28-12.76), whereas national data showed that service utilisation was associated with ownership of both records compared with owning a single record or none. CONCLUSION: Our results suggest that pre- and post-natal home-based record use may be effective for ensuring service utilisation. In addition, since the handbook is an efficient home-based record for use throughout children's life courses, it could be an effective tool for promoting the continuum of MNCH care in Indonesia.

The medial preoptic area modulates cocaine-induced activity in female rats.

Drugs of abuse exert their effects by exploiting natural neurobiological reward mechanisms, especially the mesolimbic dopamine (DA) system. However, the mesolimbic system does not operate in isolation, and input from other reward-relevant structures may play a role in cocaine's rewarding effects. The medial preoptic area (mPOA) of the hypothalamus is involved in the regulation of two essential and naturally rewarding behaviors: sexual and maternal behaviors. It also makes strong neuroanatomical connections with areas of the mesolimbic system, particularly the ventral tegmental area (VTA). As such, the mPOA is a logical candidate for a neuroanatomical locus modulating activity in the mesolimbic system and emergent behavioral expressions of drug reward, yet the role of this structure is largely unexplored. Here, using a female rat model, we show that the mPOA innervates the VTA in a region-specific manner, that lesions of the mPOA augment cocaine-induced Fos expression in the nucleus accumbens (NAc) and cocaine-induced conditioned place preference. We also show that approximately 68% of mPOA-VTA efferents release γ-aminobutyric acid (GABA), over 75% are sensitive to DA as evidenced by colocalization with DA receptors, and nearly 60% of these contain both DA receptors and GABA, which suggests a novel key role for the mPOA in the inhibition of the mesolimbic DA circuit. Combined, these results reveal the mPOA as a critical modulating structure in cocaine-induced mesolimbic activity and behavioral manifestation of reward, at least in part, via GABAergic output that is sensitive to DA input.

Mating-relevant olfactory stimuli activate the rat brain in an age-dependent manner.

Chemosensory stimulation is vital for the expression of rodent sexual behavior. As sexual activity decreases with aging, this study investigated whether aging also impacts the integration of sex-relevant chemosensory cues. To this end, several measures were obtained from adult (10-12 months) and aged (30-36 months) male rats after exposure to a conspecific estrous female. These included rates of investigatory behaviors, levels of stimulus-induced Fos immunoreactivity, activation of gonadotropin-releasing hormone-containing cells, and levels of circulating testosterone and corticosterone. The results indicated no significant differences in investigatory behaviors, levels of corticosterone, or activation of gonadotropin-releasing hormone-containing cells between the two groups. As has been reported previously, the levels of testosterone were lower in the aged rats. However, stimulus-induced neural activity was higher in the bed nucleus of the stria terminalis and the medial preoptic area of aged rats, whereas no differences were found in the main olfactory bulb, accessory olfactory bulb, medial amygdala, ventral tegmental area, or nucleus accumbens. These findings suggest the presence of a compensatory mechanism in the hypothalamus of aged animals versus adults, whereby more cells are recruited to elicit a sexual response in the presence of a sexually exciting stimulus.

Comparison of arginine vasotocin immunoreactivity differences in dominant and subordinate green anole lizards.

The neuropeptide arginine vasotocin (AVT) and its mammalian homologue arginine vasopressin (AVP) are believed to be involved in many social behaviors including territorial aggression. Testosterone (T) is also important for controlling territorial aggression, and it is believed to be involved in modulating AVT/AVP levels in the brain. In this study, male Anolis carolinensis were paired (n=11 pairs) in a neutral cage and were allowed to establish a dominant-subordinate relationship for 10 days (experimental groups) or housed in a neutral cage with or without a female (control groups; each n=4). On 10th day animals were sacrificed and their brain sections were processed for AVT immunohistochemistry and their serum was analyzed for testosterone levels. AVT immunoreactive (AVT-ir) cell numbers were counted in the anterior hypothalamus (AH), paraventricular nucleus (PN), posterior hypothalamus (PH), preoptic area (POA), and supra optic nuclei (SON). 2-way randomized block design was conducted to assess AVT-ir cell number differences between dominant and subordinate animals and Pearson's correlations were used to determine if a relationship existed between T levels and AVT-ir cell numbers. Dominant animals had more AVT-ir cells in the POA compared to subordinate animals, and subordinate animals had fewer AVT-ir cells in the POA compared to males housed either singly or with a female. There were no differences in AVT-ir cell numbers between dominant and subordinate animals in other areas. T levels were not correlated with the AVT-ir cell numbers in any area. Thus dominant animals have increased AVT-ir cell numbers compared to subordinate animals in a brain region known to be important in male sexual behavior. However, this difference is not related to differences in T.