M.S.A. Kumar

Effect of Δ9-THC on brain and plasma catecholamine levels as measured by HPLC

The effects of chronic administration of delta 9-tetrahydrocannabinol (delta 9-THC) on the plasma and brain catecholamine (CA) levels were measured using high performance liquid chromatography-electrochemical detection (LC-EC) system. Intact male rats were injected daily with vehicle (50 microliter oil) or with delta 9-THC (3 mg/kg body wt) over a period of 25 days. Trunk plasma and tissue from preoptic area (POA) and mediobasal hypothalamus (MBH) were collected and catecholamine levels were detected by LC-EC system coupled to an electronic integrator. Alumina extract of tissue and plasma samples, spiked with the internal standard (dihydroxybenzylamine), were injected into the LC-EC system; the CA were chromatographed and eluted within 12 minutes using sodium phosphate buffer as the mobile phase. delta 9-THC treatment resulted in a significant decrease in plasma and MBH levels of norepinephrine (NE), epinephrine (E), POA levels of NE; and significant increases in MBH levels of dopamine (DA) and dihydroxyphenylacetic acid (DOPAC). Our study indicates for the first time that delta 9-THC treatment significantly alters not only the POA and MBH CA levels, but also the plasma CA levels.

Substance P and Luteinizing Hormone-Releasing Hormone Levels in the Brain of the Male Golden Hamster Are Both Altered by Castration and Testosterone Replacement

The effects of castration and testosterone (T) replacement on levels of substance P (SP) and luteinizing hormone-releasing hormone (LHRH) were assessed in discrete areas of the male hamster brain. The animals were either castrated, castrated and given a chronically low or high dose of T by Silastic implant, or sham-operated. Brain tissues and trunk blood were collected 3 weeks after surgery. Plasma T levels were maintained within the normal range by the implants but at significantly lower or higher levels than the mean for sham-operated males. Levels of SP and LHRH were quantified in the olfactory bulbs, rostral basal forebrain, anterior hypothalamic and preoptic area, medial basal hypothalamic area, medial basal hypothalamic area and median eminence, and brain stem. In general, castration and T replacement effected opposite changes in levels of SP and LHRH. In the medial basal hypothalamic area and median eminence SP levels were found to be inversely related to the chronic T levels, whereas the LHRH levels were directly correlated. In the anterior hypothalamic and preoptic area, castration reduced levels of SP. Conversely, castration elevated levels of LHRH in this area. This inverse dynamic relationship between changing peptide levels was also observed in the rostral basal forebrain but not in the olfactory bulbs. In most of these forebrain regions, the dose-response curves for the experimental groups could not incorporate the peptide levels in the sham-operated control group. SP levels in the brain stem showed a monotonic inverse relationship to circulating T levels which did include the control group values.(ABSTRACT TRUNCATED AT 250 WORDS)

Distribution of substance P, GnRh, met-enkephalin in the central nervous system of the pig

The distribution of substance P (SP), gonadotropin releasing hormone (GnRH) and Met-enkephalin in the brain and spinal cord of the domestic pig is described for the first time. The levels of SP, GnRH and Met-enkephalin were measured by specific radioimmunoassays in various regions of the brain and spinal cord of the pig. Substance P and Met-enkephalin are widely distributed within the central nervous system of the pig. High levels of SP were found in the preoptic area (POA), suprachiasmatic area (SCA), medial basal hypothalamus (MBH) and brain stem while moderate amounts of SP were found in olfactory bulb (OB). High levels of Met-enkephalin were found in POA, SCA and MBH, and moderate levels of Met-enkephalin in OB and brain stem. Both SP and Met-enkephalin levels were higher in the dorsal spinal cord in comparison with the levels of these peptides in the ventral spinal cord. This finding is in agreement with the predominant role played by these neural systems in primary afferent mediation of nociceptive impulses. The POA and SCA contained only low levels of GnRH while the MBH contained high levels of GnRH. Finally, some differences in the quantitative distribution of these peptides in the pig and rat are discussed.

Effects of halothane and methoxyflurane on the hypothalamic-pituitary-adrenal axis in rat

Effects of acute exposure (2 h) to either 1.5% halothane or 0.5% methoxyflurane on chemical mediators of the hypothalamic-pituitary-adrenal (HPA) axis were evaluated in male Sprague-Dawley rats immediately after exposure, after the righting reflex (4 h), or 24 h postexposure. Effects of these anesthetics on hippocampal corticotropin releasing factor (CRF) were also evaluated. Methoxyflurane caused significant elevations in pituitary adrenocorticotropin hormone (ACTH)-like immunoreactivities in all three of the experiments time groups, yet halothane failed to cause the same response immediately after exposure. Serum ACTH-like immunoreactivities were significantly elevated immediately after exposure to both anesthetics, but were not elevated at 4 and 24 h postexposure. Corticosterone (CORT)-like immunoreactivities were significantly elevated by halothane in all experimental groups, and in the 2- and 24-h groups following methoxyflurane exposure. Hippocampal CRF-like immunoreactivities remained unaffected by either anesthetic. Results indicate that a 2-h exposure to either halothane or methoxyflurane results in significant activation of the rat hypothalamic-pituitary-adrenal axis, and that the activation appears to be sustained over a 24-h period.

Lead toxicosis in cats—a review

Although the incidence of lead toxicosis in small animals continues to decrease, it remains a significant malady. We have reviewed the literature of the past 45 years, which revealed 70 cases involving cats. Sources, signs, diagnosis, pathology and treatment of feline lead toxicosis are reviewed. In 84% of these cases the source of lead was old paint usually from home renovation. The most common signs in cats are anorexia, vomiting, and seizures. The younger individuals seem more likely to show CNS signs. Since signs are often vague, lead toxicosis may be significantly under diagnosed in cats. The gold standard of diagnostic tests is blood lead concentration, although it does not necessarily correlate with total body burden of lead or with metabolic effects including clinical signs. Diagnostic tests including erythropoietic protoporphyrin (EPP), urine aminolevulinic acid, and others are discussed. Gross findings on necropsy are few and include a yellow-brown discoloration of the liver often with a nutmeg-like appearance. Histological examination may reveal pathognomonic inclusion bodies in liver and renal tissues. Characteristic histological changes in the CNS include neuronal necrosis and demyelination. Treatment of lead toxicosis in cats, as in any species, involves removing the exposure, decontaminating the individual and the environment, supportive care and chelation therapy. The most recently available chelator is succimer (meso 2,3-dimercaptosuccinic acid). Succimer given orally is well tolerated and has a wide margin of safety. A high index of suspicion of lead toxicosis is warranted in cats since they often present with vague and non-specific signs. With any consistent history owners need to be asked about home renovation. Early diagnosis and treatment affords a good prognosis.

Distribution of catecholamines in the central nervous system of the pig

The objective of this study was to document, through comprehensive means, normal distribution and concentration of catecholamines in various regions of the CNS of pigs, an increasingly popular animal model used for transgenic manipulation of neural genes. The effects of gonadal steroidal status on this distribution were also assessed by comparing CNS catecholamine concentrations among mature male pigs (boars), immature (gilts) and mature female pigs (sows), and adult male pigs castrated prepuberally (barrows). Dissected tissue samples from the CNS were extracted in 2 N acetic acid, filtered through a 0.2 micron filter, then quantitated by reverse-phase high performance liquid chromatography using a C-18 reverse phase column with electrochemical detection. In both boars and sows the highest concentrations of norepinephrine (NE) were found in the diencephalic areas and brain stem. Gilts exhibited elevated concentrations of NE in the olfactory bulbs (OB), hypothalamus, pons, and corpus trapezoideum-locus ceruleus (LC) compared to lower concentrations in corresponding areas of sows. Prepuberal castration of the male was associated with significantly lower NE concentrations in the striatum, periaqueductal area (PAG), pons, LC, and spinal cord. The sow exhibited significantly lower NE concentrations in the mammillary area (Mam), PAG, pons, and spinal cord than those in corresponding areas of the boar. Dopamine concentrations appeared to be similar in all areas of the brain and spinal cord studied in the sow and boar. Results demonstrated that prepuberal castration of the male appears to significantly alter the DA content of the Mam and dorsal spinal cord, in contrast to gilts who possess significantly higher concentrations of DA. It is concluded from our studies that in general, catecholamine concentrations in various regions of the brain and spinal cord of sexually mature pigs parallel distributions of neuropeptides, substance P, and methionine enkephalin, as previously reported. In addition, significant association was found between gonadal activity and catecholamine concentrations in discrete areas of the pig brain.

Alterations in Catecholamine Turnover in Specific Regions of the Rat Brain Following Acute Exposure to Nitrous Oxide

The effects of nitrous oxide (N2O) on steady-state concentrations and turnover rates of catecholamines in the olfactory bulb, hypothalamus, brain stem, hippocampus, striatum, thalamus, cerebral cortex, and spinal cord were determined in rats. Animals were exposed for 2 h to either 60% N2O or air. Immediately following exposure, all animals were injected intraperitoneally with alpha-methylparatyrosine (alphaMPT), a competitive inhibitor of tyrosine hydroxylase, and sacrificed at 0, 30, or 90 min postinjection. Brain catecholamine concentrations were determined using high-performance liquid chromatography coupled with electrochemical detection (HPLC-EC). Results indicate that N2O exposure significantly elevates steady-state concentrations of norepinephrine (NE) in the hypothalamus and striatum yet decreases amine levels in the brain stem region. Steady-state levels of dopamine (DA) were not significantly altered in any region of the CNS by N2O exposure. Acute exposure to N2O also resulted in significant decreases in the turnover rate of NE in the brain stem, yet it increased turnover of this amine in the olfactory bulb, hypothalamus, and striatum. Acute exposure to N2O resulted in a decreased turnover rate of DA in the hippocampus and striatum. In contrast, N2O appears to increase DA turnover in the olfactory bulb. These results indicate that acute exposure to N2O in rats causes region-specific alterations in steady-state levels and turnover rates of DA and NE within the central nervous system.

Effects of Halothane and Methoxyflurane on Regional Brain and Spinal Cord Substance P-Like and Beta-Endorphin-Like Immunoreactivities in the Rat

Effects of acute exposure (2 hr) to either 1.5% halothane or 0.5% methoxyflurane were investigated in the Sprague Dawley rat. Pituitary (PIT) and central nervous system (CNS) substance P (SP)-like and beta-endorphin (beta-end)-like immunoreactivities were evaluated immediately after anesthetic exposure (2 h), after righting reflex (4 h) or 24 hr postexposure (24 h). Only halothane significantly reduced SP-like immunoreactivity in olfactory bulbs in both the 2-h and 4-h groups. Halothane elevated SP-like immunoreactivity of hippocampus at all three time periods, and in the hypothalamus at 2 h. Both anesthetics significantly depleted thalamic concentrations of SP-like immunoreactivity. Methoxyflurane anesthesia resulted in a drastic decrease in SP-like immunoreactivity in PIT at all three time periods periods, while halothane elevated PIT concentrations of this peptide at 4 h. Both anesthetics significantly decreased beta-end-like immunoreactivity in the olfactory bulbs and thalami at 2, 4, and 24 h. However, halothane alone significantly elevated beta-end-like immunoreactivity in the spinal cord at 24 h. Halothane significantly elevated PIT beta-end-like immunoreactivity at 2 and 24 h, while methoxyflurane significantly lowered it in the 4-h group, but elevated the levels of the same in the 24-h group. Brain stem beta-end immunoreactivity were significantly reduced at 2 h by both anesthetics, and at 4 h by methoxyflurane. Results indicate that halothane and methoxyflurane may differ significantly in their actions on SP and beta-end secreting neurons in the CNS.

Regional distribution of gonadotropin-releasing hormone-like, β-endorphin-like, and methionine-enkephalin-like immunoreactivities in the central nervous system of the goat

Regional distribution of gonadotropin-releasing hormone (GnRH)-like-, beta-endorphin (beta-end)-like-, and methionine-enkephalin (met-enk)-like-immunoreactivity was quantified across various regions of the central nervous system (CNS) of male and female goats by using highly specific radioimmunoassays. All the animals were sacrificed during the months of March through June (non-breeding season). Although the distribution of these three neuropeptides was similar to other mammalian species, species-specific gender differences in the levels of neuropeptides were noticed in the goat CNS. Highest levels of GnRH-like immunoreactivities were found in the hypothalamus. The hypothalamus of male goats exhibited significantly higher levels of GnRH-like immunoreactivities compared to female goats. Other regions exhibiting GnRH-like immunoreactivities included olfactory bulbs, preoptic and supraoptic regions, and mamillary bodies. Both beta-end- and met-enk immunoreactivities were detected in all selected regions of goat CNS, but highest levels of these opioid peptide-like immunoreactivities were limited to the forebrain regions of the goat. The supraoptic area of the female goats contained significantly higher levels of beta-end-like immunoreactivities than that of the male goats. Met-enk-peptide-like immunoreactivity also exhibited gender-specific differences in its content in some regions of the CNS. The male goats exhibited significantly higher levels of met-enk-like immunoreactivity in both the striatal and hypothalamic regions of the brain.

Effect of midazolam infusion and flumazenil administration on epinephrine arrhythmogenicity in dogs anesthetized with halothane

BackgroundMidazolam is being selected increasingly for use in patients with cardiovascular compromise. Although clinical doses of midazolam have minimal effects on cardiac function, the influence of midazolam (and other benzodiazepine sedatives) on cardiac arrhythmogenesis has yet to be elucidated fully. MethodsIn this study, we investigated the effect of midazolam, with and without flumazenil, on the arrhythmogenic serum concentration of epinephrine (ACE) in six halothane-anesthetized dogs. Midazolam was administered as a loading dose (1.5 mg/kg over 5 min) followed by a 4.5-h infusion at two rates (10 and 40 μB. kg−1. min−1) to achieve and maintain predetermined clinical and supraclinical plasma midazolam concentrations. The arrhythmogenic serum concentration of epinephrine determinations were made prior to midazolam infusion, following 2 h of midazolam infusion and following 3.5 h of midazolam infusion and 1 mg flumazenil/kg iv. Saline control studies were also performed in four of the six dogs. ResultsPlasma midazolam concentrations ranged from 363 to 855 ng/ml in the low-dose infusion study, essentially spanning the clinically effective range for humans. In the high-dose infusion study, plasma midazolam concentrations were up to four times greater, ranging from 1168 to 3563 ng/ml. The arrhythmogenic serum concentration of epinephrine values were unchanged following low-dose midazolam infusion and saline. In the high-dose study, ACE increased from baseline values of 68 ± 13 (SEM) ng/ml to 112 ± 25 ng/ml (P=.03) following midazolam infusion and decreased to 79 ±13 ng/ml with flumazenil administration. Plasma midazolam concentrations, however, were poorly correlated with ACE values normalized for control ACE (ACE ratio). Diastolic arterial pressure was significantly depressed following both low-dose (-14%) and high-dose (-19%) midazolam infusion. This decrease in blood pressure was unaffected by flumazenil administration. Other hemodynamic parameters were unaffected by drug treatment. ConclusionsThis study has demonstrated that midazolam infusion results in either no effect (with clinical plasma midazolam concentrations) or flumazenil-reversible suppression (with supraclinical concentrations) of halothane-epinephrine arrhythmogenesis.