Wildfire Induces Changes in Receiving Waters: A Review With Considerations for Water Quality Management.

Wildfires have increased in frequency in many ecosystems, with implications for human health and the environment, including water quality. Increased fire frequency and urbanization also raise the prospect of fires burning into urban areas, mobilizing pollutants few have considered to date. As a result, water quality managers lack information to anticipate, respond to and potentially mitigate wildfire impacts. Here, we reviewed the scientific literature to assess wildfire effects on response endpoints of a conceptual model linking fire to water quality, quantifying response directionality, magnitude and duration. Physically, water yield, sediments, and temperature all increased post-fire. Chemically, nutrients, ions, organic chemicals, and metals increased in burned watersheds, sometimes by orders of magnitude over pre-fire or reference conditions. In select cases, post-fire concentrations exceeded aquatic life criteria or drinking water standards, at times even in the finished drinking water. Biological assemblages commonly declined after post-fire runoff events. The duration of effects was less than 5 yr for most endpoints (e.g., metals) on average following fire, although effects did extend 15 yr or more in some individual cases. We found only a few studies on pollutants mobilized from wildfire impacted urban areas with benzene contamination in drinking water and high metal concentrations in ash prominent exceptions. Overall, this review provides a resource for understanding wildfire impacts on water quality endpoints, with the goal of informing the response of managers and other decision makers to this growing problem.

Estimating wetland connectivity to streams in the Prairie Pothole Region: an isotopic and remote sensing approach.

Understanding hydrologic connectivity between wetlands and perennial streams is critical to understanding the reliance of stream flow on inputs from wetlands. We used the isotopic evaporation signal in water and remote sensing to examine wetland-stream hydrologic connectivity within the Pipestem Creek watershed, North Dakota, a watershed dominated by prairie-pothole wetlands. Pipestem Creek exhibited an evaporated-water signal that had approximately half the isotopic-enrichment signal found in most evaporatively enriched prairie-pothole wetlands. Groundwater adjacent to Pipestem Creek had isotopic values that indicated recharge from winter precipitation and had no significant evaporative enrichment, indicating that enriched surface water did not contribute significantly to groundwater discharging into Pipestem Creek. The estimated surface-water area necessary to generate the evaporation signal within Pipestem Creek was highly dynamic, varied primarily with the amount of discharge, and was typically greater than the immediate Pipestem Creek surface-water area, indicating that surficial flow from wetlands contributed to stream flow throughout the summer. We propose a dynamic range of spilling thresholds for prairie-pothole wetlands across the watershed allowing for wetland inputs even during low flow periods. Combining Landsat estimates with the isotopic approach allowed determination of potential (Landsat) and actual (isotope) contributing areas in wetland-dominated systems. This combined approach can give insights into the changes in location and magnitude of surface water and groundwater pathways over time. This approach can be used in other areas where evaporation from wetlands results in a sufficient evaporative isotopic signal.

Short communication: Efficacy of glycolic acid-based and iodine-based postmilking barrier teat disinfectants for prevention of new intramammary infections in dairy cattle.

A positive-control, natural exposure noninferiority field study was conducted to test the efficacy of a novel glycolic acid-based postmilking barrier teat disinfectant compared with a commercial iodine-based postmilking barrier teat disinfectant (positive control). Cows from 2 pens from a California Central Valley dairy farm were dipped after milking either with the positive-control product (PC) or the experimental product (EX) over 12 wk. New intramammary infections (NIMI) were determined by biweekly sampling of all quarters of study cows and classified as a NIMI based on somatic cell count and milk bacteriological culture results. The mean quarter-level incidence risks during a 2 wk study period were 3.50% (EX) and 4.28% (PC). The majority of NIMI were caused by coagulase-negative staphylococci, followed by non-agalactiae streptococci. The study results indicated that EX was noninferior to PC, with a 17% relative efficacy (improvement) in reducing NIMI compared with the PC group. Also, quarter somatic cell count was not affected by the postmilking teat disinfectant used. Finally, the EX product was safe in terms of teat conditioning: teat condition scores were not different between study groups. The study concluded that the glycolic acid-based experimental post-dip barrier was noninferior to the control, and could be considered a safe and effective postmilking teat disinfectant.

Randomized noninferiority study evaluating the efficacy of a postmilking teat disinfectant for the prevention of naturally occurring intramammary infections.

The objective of this study was to complete a positive-control, natural exposure, noninferiority design field study to test the efficacy of a novel glycolic acid-based postmilking teat disinfectant as compared with a previously proven iodine-based postmilking teat disinfectant (positive control). The primary outcome of interest was the effect of treatment on incidence of new intramammary infections. Secondary outcomes included the effect of treatment on prevalence of infection, somatic cell count, and teat condition. After blocking by parity, approximately 300 early- to mid-lactation cows on a large Wisconsin dairy farm were randomly assigned to 1 of 2 groups. For a 12-wk period between May and August 2014, the 2 groups were dipped after each milking with either the experimental (EX) or positive control (PC) product. Individual quarters were sampled to establish bacteriological infection status at the beginning of the study, and every 2 wk thereafter, by use of a 2-stage process evaluating somatic cell count (SCC), and then culturing milk samples only when SCC exceeded a parity-specific threshold. Teat condition scoring was completed at the beginning of the study and on wk 4, 8, and 12. Mixed logistic regression was used to evaluate the effect of treatment on dichotomous outcome measures including the odds of acquiring a new infection during a given 2-wk sampling interval (incidence), the odds for presence of infection at sampling (prevalence), and odds for a normal teat skin condition score. Mixed linear regression was used to evaluate the effect of treatment on somatic cell count. For the noninferiority analysis, the upper bound of the 95% confidence interval for the difference in new infection rate between the 2 treatments (EX - PC), had to be to the left of the critical value d (0.035) to conclude that EX was noninferior relative to PC with respect to risk for new infections. Results showed that the incidence of new infections was not different for quarters dipped with EX (3.2%) as compared with PC (4.2%). Similarly, the prevalence of infection tended to be lower for quarters dipped with EX (3.92%) as compared with PC (5.03%). No overall difference was found between treatments when evaluating somatic cell count measures and teat condition scores. Because the upper bound of the 95% confidence interval of the new IMI rate difference was smaller than the predefined noninferiority limit, it was concluded that the experimental product was not inferior compared with the positive control. As such, the glycolic acid-based teat disinfectant evaluated in this study can be considered an effective postmilking teat disinfectant, as well as safe, in so far as the product was not irritating to teat skin and did not negatively affect skin condition measures, as compared with the positive control group.

Standards of Care for the Health of Transgender and Gender Diverse People, Version 8.

Background: Transgender healthcare is a rapidly evolving interdisciplinary field. In the last decade, there has been an unprecedented increase in the number and visibility of transgender and gender diverse (TGD) people seeking support and gender-affirming medical treatment in parallel with a significant rise in the scientific literature in this area. The World Professional Association for Transgender Health (WPATH) is an international, multidisciplinary, professional association whose mission is to promote evidence-based care, education, research, public policy, and respect in transgender health. One of the main functions of WPATH is to promote the highest standards of health care for TGD people through the Standards of Care (SOC). The SOC was initially developed in 1979 and the last version (SOC-7) was published in 2012. In view of the increasing scientific evidence, WPATH commissioned a new version of the Standards of Care, the SOC-8. Aim: The overall goal of SOC-8 is to provide health care professionals (HCPs) with clinical guidance to assist TGD people in accessing safe and effective pathways to achieving lasting personal comfort with their gendered selves with the aim of optimizing their overall physical health, psychological well-being, and self-fulfillment. Methods: The SOC-8 is based on the best available science and expert professional consensus in transgender health. International professionals and stakeholders were selected to serve on the SOC-8 committee. Recommendation statements were developed based on data derived from independent systematic literature reviews, where available, background reviews and expert opinions. Grading of recommendations was based on the available evidence supporting interventions, a discussion of risks and harms, as well as the feasibility and acceptability within different contexts and country settings. Results: A total of 18 chapters were developed as part of the SOC-8. They contain recommendations for health care professionals who provide care and treatment for TGD people. Each of the recommendations is followed by explanatory text with relevant references. General areas related to transgender health are covered in the chapters Terminology, Global Applicability, Population Estimates, and Education. The chapters developed for the diverse population of TGD people include Assessment of Adults, Adolescents, Children, Nonbinary, Eunuchs, and Intersex Individuals, and people living in Institutional Environments. Finally, the chapters related to gender-affirming treatment are Hormone Therapy, Surgery and Postoperative Care, Voice and Communication, Primary Care, Reproductive Health, Sexual Health, and Mental Health. Conclusions: The SOC-8 guidelines are intended to be flexible to meet the diverse health care needs of TGD people globally. While adaptable, they offer standards for promoting optimal health care and guidance for the treatment of people experiencing gender incongruence. As in all previous versions of the SOC, the criteria set forth in this document for gender-affirming medical interventions are clinical guidelines; individual health care professionals and programs may modify these in consultation with the TGD person.

Unexpected adrenergic effects of chlorpromazine: eating elicited by injection into rat hypothalamus.

Although chlorpromazine is believed to block adrenergic transmission, injection of this drug into the hypothalamus of satiated rats does not block norepinephrine-elicited eating, but instead mimics norepinephrine by eliciting eating. The amount of eating elicited by norepinephrine and by chlorpromazine is reliably correlated. These results suggest that endogenous norepinephrine mediates eating elicited by centrally injected chlorpromazine.

Amnesia or reversal of forgetting by anticholinesterase, depending simply on time of injection.

The effect of intracerebral injections of the anticholinesterase drug diisopropyl fluorophosphate in rats was to produce good recall of an otherwise almost forgotten habit learned 28 days before. The same injections produced temporary amnesia for the same habit, otherwise well remembered, learned 14 days before. The injections had no ef fect on the memory of the same habit when it was only partly learned 14 days before. The results support the hy pothesis that the physiological basis of memory lies in an increase, and for getting in a decrease, in synaptic con ductance.

Increased caloric intake on a fat-rich diet: role of ovarian steroids and galanin in the medial preoptic and paraventricular nuclei and anterior pituitary of female rats.

Previous studies in male rats have demonstrated that the orexigenic peptide galanin (GAL), in neurones of the anterior parvocellular region of the paraventricular nucleus (aPVN) projecting to the median eminence (ME), is stimulated by consumption of a high-fat diet and may have a role in the hyperphagia induced by fat. In addition to confirming this relationship in female rats and distinguishing the aPVN-ME from other hypothalamic areas, the present study identified two additional extra-hypothalamic sites where GAL is stimulated by dietary fat in females but not males. These sites were the medial preoptic nucleus (MPN), located immediately rostral to the aPVN, and the anterior pituitary (AP). The involvement of ovarian steroids, oestradiol (E(2)) and progesterone (PROG), in this phenomenon was suggested by an observed increase in circulating levels of these hormones and GAL in MPN and AP with fat consumption and an attenuation of this effect on GAL in ovariectomised (OVX) rats. Furthermore, in the same four areas affected by dietary fat, levels of GAL mRNA and peptide immunoreactivity were stimulated by E(2) and further by PROG replacement in E(2)-primed OVX rats and were higher in females compared to males. Because both GAL and PROG stimulate feeding, their increase on a fat-rich diet may have functional consequences in females, possibly contributing to the increased caloric intake induced by dietary fat. This is supported by the findings that PROG administration in E(2)-primed OVX rats reverses the inhibitory effect of E(2) on total caloric intake while increasing voluntary fat ingestion, and that female rats with higher GAL exhibit increased preference for fat compared to males. Thus, ovarian steroids may function together with GAL in a neurocircuit, involving the MPN, aPVN, ME and AP, which coordinate feeding behaviour with reproductive function to promote consumption of a fat-rich diet at times of increased energy demand.

Increased caloric intake after a high-fat preload: relation to circulating triglycerides and orexigenic peptides.

To investigate mechanisms that mediate the greater food intake induced by a fat-rich diet, the present study tested an acute "preload-to-test meal" paradigm in normal-weight rats. In this paradigm, the rats were given a small high-fat (HF) compared to low-fat (LF) preload and, after an intermeal interval, allowed to consume freely on a subsequent test meal. Modified versions of this paradigm were tested to determine the robustness of the greater caloric intake induced by the HF preload while standardizing the test protocol. A HF preload of 10-15 kcals, compared to an equicaloric LF preload, significantly increased food intake by 40-50% in the subsequent test meal. This effect, a 4-6 kcal increase, was observed with HF preloads equal in energy density and palatability to the LF preloads. It was evident with preloads or test meals that were liquid or solid, preloads that were injected, test meals that had variable fat content, and natural intermeal intervals of 60-120 min. This overeating after a HF preload was invariably associated with a 2- to 3-fold increase in circulating levels of triglycerides (TG), with no change in leptin or insulin. It was also accompanied by increased expression of the orexigenic peptides, galanin in the paraventricular nucleus and orexin in the perifornical lateral hypothalamus. Moreover, if given repeatedly over several days, the HF compared to equicaloric LF preload significantly increased 24-h food intake. These results establish a protocol for studying the phenomenon of increased feeding on a HF diet under controlled conditions and suggest possible underlying mechanisms involving circulating lipids and orexigenic peptides.

Neurochemical-neuroendocrine systems in the brain controlling macronutrient intake and metabolism.

Appetite, energy balance and body weight gain are modulated by diverse neurochemical and neuroendocrine signals from different organs in the body and diverse regions in the brain. The hypothalamus plays an important integrative function in this process, acting through a variety of systems that involve a close interaction between nutrients, amines, neuropeptides and hormones. These systems underlie normal nutrient intake and metabolism and are thought to be responsible for shifts in feeding behavior across the circadian cycle and fluctuations relating to gender and age in both rats and humans. Moreover, alterations in these normal neurochemical-neuroendocrine systems may be associated with abnormal eating patterns, such as anorexia nervosa, bulimia and obesity. Understanding the systems that control eating behavior might provide a foundation for the treatment and possible prevention of such disorders.

Model for predicting and phenotyping at normal weight the long-term propensity for obesity in Sprague-Dawley rats.

Tests were conducted to determine whether weight gain or nutrient intake measures during the first week of exposure to a macronutrient diet can accurately predict an animal's long-term propensity towards obesity. In multiple groups of normal-weight Sprague-Dawley rats (n=35-70/group), daily weight gain during the first 5 days on a high-fat diet (45-60% fat) was found to be strongly, positively correlated (r=+0.71 to r=+0.82) with accumulated body fat in 4 dissected depots after 4-6 weeks on the diet. This measure consistently identified obesity-prone (OP) rats which, relative to the obesity-resistant (OR) rats, were only slightly heavier (+15 g, 4%) and hyperphagic (+9 kcal, 8%) after 5 days but markedly heavier (+70g) with up to 2-fold greater fat mass after several weeks on the diet. Other dietary conditions and measures revealed weaker relationships to ultimate body fat accrual. The OP rats identified by their 5-day weight-gain score exhibited at this early stage clear disturbances characteristic of markedly obese rats. These included elevated leptin, insulin, triglycerides and glucose, along with increased lipoprotein lipase activity (LPL) in adipose tissue and galanin expression in the paraventricular nucleus. Most notable were significant reductions in muscle of LPL activity and ratio of beta-hydroxyacyl-CoA dehydrogenase to citrate synthase activity, indicating a decline in lipid transport and capacity of muscle to metabolize lipids. By occurring early with initial weight gain, these hypothalamic and metabolic disturbances in OP rats, favoring fat storage in adipose tissue over fat oxidation in muscle, may have causal relationships to long-term accumulation of body fat.

Prenatal fat-rich diet exposure alters responses of embryonic neurons to the chemokine, CCL2, in the hypothalamus.

Maternal consumption of a high-fat diet (HFD) during pregnancy is found to stimulate the genesis of hypothalamic orexigenic peptide neurons in the offspring, while HFD intake in adult animals produces a systemic low-grade inflammation which increases neuroimmune factors that may affect neurogenesis and neuronal migration. Building on this evidence and our recent study showing that the inflammatory chemokine, CCL2, stimulates the migration of hypothalamic neurons and expression of orexigenic neuropeptides, we tested here the possibility that prenatal exposure to a HFD in rats affects this chemokine system, both CCL2 and its receptors, CCR2 and CCR4, and alters its actions on hypothalamic neurons, specifically those expressing the neuropeptides, enkephalin (ENK) and galanin (GAL). Using primary dissociated hypothalamic neurons extracted from embryos on embryonic day 19, we found that prenatal HFD exposure compared to chow control actually reduces the expression of CCL2 in these hypothalamic neurons, while increasing CCR2 and CCR4 expression, and also reduces the sensitivity of hypothalamic neurons to CCL2. The HFD abolished the dose-dependent, stimulatory effect of CCL2 on the number of migrated neurons and even shifted its normal stimulatory effect on migrational velocity and distance traveled by control neurons to an inhibition of migration. Further, it abolished the dose-dependent, stimulatory effect of CCL2 on neuronal expression of ENK and GAL. These results demonstrate that prenatal HFD exposure greatly disturbs the functioning of the CCL2 chemokine system in embryonic hypothalamic neurons, reducing its endogenous levels and ability to promote the migration of neurons and their expression of orexigenic peptides.

Prenatal fat exposure and hypothalamic PPAR ß/δ: Possible relationship to increased neurogenesis of orexigenic peptide neurons.

Gestational exposure to a fat-rich diet, while elevating maternal circulating fatty acids, increases in the offspring's hypothalamus and amygdala the proliferation and density of neurons that express neuropeptides known to stimulate consummatory behavior. To understand the relationship between these phenomena, this study examined in the brain of postnatal offspring (day 15) the effect of prenatal fat exposure on the transcription factor, peroxisome proliferator-activated receptor (PPAR) ß/δ, which is sensitive to fatty acids, and the relationship of PPAR ß/δ to the orexigenic neuropeptides, orexin, melanin-concentrating hormone, and enkephalin. Prenatal exposure to a fat-rich diet compared to low-fat chow increased the density of cells immunoreactive for PPAR ß/δ in the hypothalamic paraventricular nucleus (PVN), perifornical lateral hypothalamus (PFLH), and central nucleus of the amygdala (CeA), but not the hypothalamic arcuate nucleus or basolateral amygdaloid nucleus. It also increased co-labeling of PPAR ß/δ with the cell proliferation marker, BrdU, or neuronal marker, NeuN, and the triple labeling of PPAR ß/δ with BrdU plus NeuN, indicating an increase in proliferation and density of new PPAR ß/δ neurons. Prenatal fat exposure stimulated the double-labeling of PPAR ß/δ with orexin or melanin-concentrating hormone in the PFLH and enkephalin in the PVN and CeA and also triple-labeling of PPAR ß/δ with BrdU and these neuropeptides, indicating that dietary fat increases the genesis of PPAR ß/δ neurons that produce these peptides. These findings demonstrate a close anatomical relationship between PPAR ß/δ and the increased proliferation and density of peptide-expressing neurons in the hypothalamus and amygdala of fat-exposed offspring.

Effects of embryonic ethanol exposure at low doses on neuronal development, voluntary ethanol consumption and related behaviors in larval and adult zebrafish: Role of hypothalamic orexigenic peptides.

Embryonic exposure to ethanol is known to affect neurochemical systems in rodents and increase alcohol drinking and related behaviors in humans and rodents. With zebrafish emerging as a powerful tool for uncovering neural mechanisms of numerous diseases and exhibiting similarities to rodents, the present report building on our rat studies examined in zebrafish the effects of embryonic ethanol exposure on hypothalamic neurogenesis, expression of orexigenic neuropeptides, and voluntary ethanol consumption and locomotor behaviors in larval and adult zebrafish, and also effects of central neuropeptide injections on these behaviors affected by ethanol. At 24h post-fertilization, zebrafish embryos were exposed for 2h to ethanol, at low concentrations of 0.25% and 0.5%, in the tank water. Embryonic ethanol compared to control dose-dependently increased hypothalamic neurogenesis and the proliferation and expression of the orexigenic peptides, galanin (GAL) and orexin (OX), in the anterior hypothalamus. These changes in hypothalamic peptide neurons were accompanied by an increase in voluntary consumption of 10% ethanol-gelatin and in novelty-induced locomotor and exploratory behavior in adult zebrafish and locomotor activity in larvae. After intracerebroventricular injection, these peptides compared to vehicle had specific effects on these behaviors altered by ethanol, with GAL stimulating consumption of 10% ethanol-gelatin more than plain gelatin food and OX stimulating novelty-induced locomotor behavior while increasing intake of food and ethanol equally. These results, similar to those obtained in rats, suggest that the ethanol-induced increase in genesis and expression of these hypothalamic peptide neurons contribute to the behavioral changes induced by embryonic exposure to ethanol.

PVN galanin increases fat storage and promotes obesity by causing muscle to utilize carbohydrate more than fat.

To understand the function of the feeding-stimulatory peptide, galanin (GAL), in eating and body weight regulation, the present experiments tested the effects of both acute and chronic injections of this peptide into the paraventricular nucleus (PVN) of rats. With food absent during the test, acute injection of GAL (300 pmol/0.3 microl) significantly increased phosphofructokinase activity in muscle, suggesting enhanced capacity to metabolize carbohydrate, and reduced circulating glucose levels. It also decreased beta-hydroxyacyl-CoA dehydrogenase activity in muscle, indicating reduced fat oxidation, while increasing circulating non-esterified fatty acids (NEFA) and lipoprotein lipase activity in adipose tissue (aLPL). Chronic PVN injections of GAL (300 pmol/0.3 microl/injection) versus saline over 7-10 days significantly stimulated daily caloric intake and increased the weight of four dissected fat depots by 30-40%. These effects, accompanied by elevated levels of leptin, triglycerides, NEFA and aLPL activity, were evident only in rats on a diet with at least 35% fat. Thus, by favoring carbohydrate over fat metabolism in muscle and reversing hyperglycemia, PVN GAL may have a function in counteracting the metabolic disturbances induced by a high-fat diet. As a consequence of these actions, GAL can promote the partitioning of lipids away from oxidation in muscle towards storage in adipose tissue.

Galanin and alcohol dependence: neurobehavioral research.

It is known that microinjection of galanin (GAL) intraventricularly or in specific hypothalamic sites increases food consumption and, conversely, the intake of food increases the expression of GAL in hypothalamic sites. Ethanol (EtOH) is a calorie-rich food as well as a drug of abuse. The research reviewed here shows that GAL may play a similar role in alcohol intake. First, experiments in which GAL was microinjected into the third ventricle or the paraventricular nucleus (PVN) showed increases in EtOH consumption. The increase in EtOH consumption occurred during both the light and dark cycles after GAL injection in the third ventricle in rats with limited EtOH access. Injection of GAL did not increase food intake in rats that had been chronically drinking alcohol. GAL receptor blockade reversed these increases. Microinjection of GAL directly into the PVN also increased ad libitum EtOH intake and blockade of these receptors in the PVN inhibited ad libitum EtOH consumption. Secondly, rats administered EtOH showed increases in GAL in the PVN and related hypothalamic sites. EtOH injection and voluntary intake, both ad libitum and limited access, increased GAL gene and peptide expression in the PVN consistently across administration procedures. These experiments show that GAL injection increases alcohol intake and that the intake of alcohol increases GAL, suggesting a positive feedback relationship between alcohol intake and specific hypothalamic GAL systems. Such a relationship may contribute to the motivation to consume excessive alcoholic beverages and the development of alcohol dependence.

Prenatal exposure to ethanol stimulates hypothalamic CCR2 chemokine receptor system: Possible relation to increased density of orexigenic peptide neurons and ethanol drinking in adolescent offspring.

Clinical and animal studies indicate that maternal consumption of ethanol during pregnancy increases alcohol drinking in the offspring. Possible underlying mechanisms may involve orexigenic peptides, which are stimulated by prenatal ethanol exposure and themselves promote drinking. Building on evidence that ethanol stimulates neuroimmune factors such as the chemokine CCL2 that in adult rats is shown to colocalize with the orexigenic peptide, melanin-concentrating hormone (MCH) in the lateral hypothalamus (LH), the present study sought to investigate the possibility that CCL2 or its receptor CCR2 in LH is stimulated by prenatal ethanol exposure, perhaps specifically within MCH neurons. Our paradigm of intraoral administration of ethanol to pregnant rats, at low-to-moderate doses (1 or 3g/kg/day) during peak hypothalamic neurogenesis, caused in adolescent male offspring twofold increase in drinking of and preference for ethanol and reinstatement of ethanol drinking in a two-bottle choice paradigm under an intermittent access schedule. This effect of prenatal ethanol exposure was associated with an increased expression of MCH and density of MCH(+) neurons in LH of preadolescent offspring. Whereas CCL2(+) cells at this age were low in density and unaffected by ethanol, CCR2(+) cells were dense in LH and increased by prenatal ethanol, with a large percentage (83-87%) identified as neurons and found to colocalize MCH. Prenatal ethanol also stimulated the genesis of CCR2(+) and MCH(+) neurons in the embryo, which co-labeled the proliferation marker, BrdU. Ethanol also increased the genesis and density of neurons that co-expressed CCR2 and MCH in LH, with triple-labeled CCR2(+)/MCH(+)/BrdU(+) neurons that were absent in control rats accounting for 35% of newly generated neurons in ethanol-exposed rats. With both the chemokine and MCH systems believed to promote ethanol consumption, this greater density of CCR2(+)/MCH(+) neurons in the LH of preadolescent rats suggests that these systems function together in promoting alcohol drinking during adolescence.

Model of voluntary ethanol intake in zebrafish: effect on behavior and hypothalamic orexigenic peptides.

Recent studies in zebrafish have shown that exposure to ethanol in tank water affects various behaviors, including locomotion, anxiety and aggression, and produces changes in brain neurotransmitters, such as serotonin and dopamine. Building on these investigations, the present study had two goals: first, to develop a method for inducing voluntary ethanol intake in individual zebrafish, which can be used as a model in future studies to examine how this behavior is affected by various manipulations, and second, to characterize the effects of this ethanol intake on different behaviors and the expression of hypothalamic orexigenic peptides, galanin (GAL) and orexin (OX), which are known in rodents to stimulate consumption of ethanol and alter behaviors associated with alcohol abuse. Thus, we first developed a new model of voluntary intake of ethanol in fish by presenting this ethanol mixed with gelatin, which they readily consume. Using this model, we found that individual zebrafish can be trained in a short period to consume stable levels of 10% or 20% ethanol (v/v) mixed with gelatin and that their intake of this ethanol-gelatin mixture leads to pharmacologically relevant blood ethanol concentrations which are strongly, positively correlated with the amount ingested. Intake of this ethanol-gelatin mixture increased locomotion, reduced anxiety, and stimulated aggressive behavior, while increasing expression of GAL and OX in specific hypothalamic areas. These findings, confirming results in rats, provide a method in zebrafish for investigating with forward genetics and pharmacological techniques the role of different brain mechanisms in controlling ethanol intake.

Hypothalamic paraventricular nucleus: interaction between alpha 2-noradrenergic system and circulating hormones and nutrients in relation to energy balance.

Extensive evidence suggests that norepinephrine (NE) in the brain is active in the control of eating behavior. Central injection studies demonstrate a stimulatory effect of NE on food intake, a response which is mediated by alpha 2-noradrenergic receptors located in the medial hypothalamus, in particular the paraventricular nucleus (PVN). Activation of these PVN receptors stimulates ingestion specifically of carbohydrate-rich foods, and this response is believed to reflect the role of endogenous NE in controlling natural appetite for this macronutrient. This alpha 2-noradrenergic system in the PVN appears to be physiologically activated at the onset of the animals' active cycle, when there is a natural peak in preference for carbohydrate. At this time, the adrenal hormone corticosterone, which is known to play a major role in carbohydrate metabolism, is found to interact positively with NE in the potentiation of carbohydrate ingestion. Circulating glucose also influences the activity of PVN alpha 2-noradrenergic receptors at this time, and, moreover, alpha-noradrenergic stimulation of the PVN produces an increase in circulating levels of both corticosterone and glucose. This and other evidence has led to the hypothesis that NE in the PVN, through the activation of glucocorticoid- and glucose-sensitive alpha 2-receptor sites, is physiologically active in energy homeostasis, most particularly at the onset of the animal's active cycle. Specifically, this neurotransmitter in the PVN evokes a state of energy conservation. This state involves adjustments in carbohydrate ingestion as well as metabolism, that allow animals to maintain energy reserves by anticipating or responding to a depletion.

Gonadal steroids and hypothalamic galanin and neuropeptide Y: role in eating behavior and body weight control in female rats.

The neuropeptides, galanin (GAL) and neuropeptide Y (NPY), based on studies in male rodents, are believed to have a role in controlling energy balance, both nutrient ingestion and metabolism. Whereas these peptides are also involved in reproduction, little is known about their specific function in energy balance in females. In rats consuming lab chow or macronutrient diets, measurements across the estrous cycle were taken of hypothalamic GAL and NPY, using RIA and immunohistochemistry; of the circulating hormones, estradiol, progesterone, and LH; and also of food intake and body weight. Levels of GAL and NPY peak during the proestrous phase of the female cycle when circulating estradiol and progesterone also rise. As previously reported for GAL, this peak is detected in two areas, the medial preoptic area (MPOA; +110%; P < 0.05) and the external zone of the median eminence (+57%; P < 0.05). In addition, this proestrous peak is seen in the paraventricular nucleus (PVN), specifically the anterior parvocellular portion (+35%; P < 0.05). Similarly, NPY rises during proestrous in the medial region of the PVN (+21%; P < 0.05) in addition to the MPOA (+78%; P < 0.05) and arcuate nucleus (+35%; P < 0.05). This peak in peptide levels is accompanied by an increase in caloric intake in rats receiving the lab chow diet and a specific increase in preference for fat in rats receiving macronutrient diets. Animals showing a preference for a fat-rich diet exhibit higher levels of GAL in the MPOA as well as the PVN and median eminence and also of NPY specifically in the MPOA. These peptides in the MPOA are similarly enhanced in animals with greater body fat, independent of diet. This evidence suggests that in the female rat, both GAL and NPY in the MPOA may contribute to the overeating and increased weight gain that occur during a fat-rich diet.