Effects of pinealectomy and melatonin administration on thyroid follicles of blind Syrian hamsters

Since previous studies have shown that an active pineal gland exerts an inhibitory effect on circulating levels of thyroxin in the Syrian hamster, a study was conducted to determine whether the histology and ultrastructure of the thyroid gland supported the conclusions drawn from the hormone data. The ultrastructure of thyroid glands of blinded male Syrian hamsters was compared to that of intact controls kept under a 14L/10D photoperiod, to that of blinded hamsters also pinealectomized, and to that of blinded hamsters receiving 80 micrograms/ml of melatonin in the drinking water. Serum thyroxin (T4) and serum thyrotropin (TSH) concentrations were determined by radioimmunoassay. After 10 weeks serum thyroxin concentrations were less than 50% of controls and concentrations were significantly reduced. EM examination revealed that blinded hamsters had an increased number of follicular cells with flattened epithelium and nondilated endoplasmic reticulum compared to intact controls. In blinded hamsters that were pinealectomized or treated with melatonin in the drinking water, the ultrastructure of the thyroid was not different from controls and serum thyroxin concentrations were restored to near normal. These ultrastructural data support the conclusion that the pineal gland is required to obtain inhibition of the pituitary-thyroid axis in blinded hamsters and that melatonin has a counter-inhibitory effect when administered via the drinking water.     

Effects of methimazole pretreatment on cerulein induced acute pancreatitis in rats.

BACKGROUND: Many interrelationships exist between the thyroid gland and the gastrointestinal tract. Several past and recent studies have shown that the thyroid gland profoundly influences the structure and function of the exocrine pancreas in the rat. In the present study we investigated the effect of methimazole (METZ), an antithyroid drug, on cerulein induced acute pancreatitis (AP) in rats. METHODS: Rats were divided into 3 groups (10-12 weeks age, 200-250 g weight, n: 10). Group B was made hypothyroid with methimazole 5 mg/kg daily for 10 days and the others were untreated euthyroid groups. After 10 days, acute pancreatitis was induced with four doses of 20 microg/kg body weight of cerulein administered s.c at hourly intervals in group A and B while the control group C was given 4 doses of I ml saline. Pancreas wet weight (mg), plasma amylase activity (IU/l) and pancreatic histology were used as endpoints to quantify the severity of the AP. RESULTS: Plasma tri-iodothyronine (T3) (ng/dl) and thyroxine (T4) (microg/dl) levels were significantly reduced after METZ treatment for 10 days (p < 0.01). METZ pretreatment reduced significantly the cerulein induced increase in pancreatic weight (1,205 +/- 12 mg in METZ treated AP group versus 1,617 +/- 14 mg in AP group, p < 0.05) and the rise in amylase activity (7,078 +/- 816 IU/l in METZ treated AP group versus 8,611 +/- 830 IU/l in AP group p < 0.05). CONCLUSION: METZ reduces the severity of cerulein induced AP in rats. This effect might be through its antithyroid property.     

Histamine and serotonin independently elicit drinking in the rat

Drinking elicited by SC histamine or serotonin (5-HT) was studied in Sprague-Dawley male rats following 0.9% NaCl or combined antagonism of H1 and H2 histamine receptors using dexbrompheniramine (DXB) and cimetidine (C), or following antagonism of 5-HT receptors using methysergide (M). Histaminergic antagonism using IP 1 mg/kg DXB plus 16 mg/kg C abolished drinking elicited by SC 2.5 mg/kg histamine, but it failed to inhibit drinking elicited by the ED50 or by the ED100 for SC 5-HT in the same rats. Serotonergic antagonism using IP 3 mg/kg M abolished drinking elicited by SC 0.63 mg/kg 5-HT, but it failed to inhibit drinking elicited by the ED50 or by the ED100 for SC histamine in the same rats. These findings demonstrate that activation of peripheral 5-HT receptors is not necessary for SC histamine to elicit drinking and that activation of peripheral histamine receptors is not necessary for SC 5-HT to elicit drinking. This demonstrates that histamine and 5-HT activate different receptors to elicit drinking in the rat. The finding that the ED50 for histamine and the ED50 for 5-HT are not additive in their effects on drinking is consistent with the notion that a single mechanism mediates the dipsogenic effects of SC histamine and SC 5-HT in the rat.     

Subchronic sodium chlorate exposure in drinking water results in a concentration-dependent increase in rat thyroid follicular cell hyperplasia.

Chlorine dioxide (ClO2) is an effective drinking water disinfectant, but sodium chlorate (NaClO3) has been identified as a potentially harmful disinfection by-product. Studies were performed to describe the development of thyroid lesions in animals exposed to NaClO3 in the drinking water. Male and female F344 rats and B6C3F1 mice were exposed to 0, 0.125, 0.25, 0.5, 1.0, or 2.0 g/L NaClO3 for 21 days. Additional male F344 rats were exposed to 0, 0.001. 0.01. 0.1, 1.0. or 2.0 g/L NaClO3 for 90 days. Female F344 rats were exposed to 0, 0.5, 1.0, 2.0, 4.0, or 6.0 g/L of NaClO3 for 105 days. Thyroid tissues were processed by routine methods for light microscopic examination, and follicular cell hyperplasia was diagnosed using a novel method. Thyroid hormone levels were altered significantly after 4 and 21 days. NaClO, treatment induced a concentration-dependent increase in the incidence and severity of thyroid follicular cell hyperplasia. Male rats are more sensitive to the effects of NaClO3 treatment than females. Follicular cell hyperplasia was not present in male or female B6C3F1 mice. These data can be used to estimate the human health risk that would be associated with using ClO2, rather than chlorine, to disinfect drinking water.     

NSE在慢性砷中毒大鼠海马CA3区的表达

目的:利用神经元特异性烯醇化酶(NSE)免疫组化染色,光镜下观察慢性砷中毒对成年大鼠海马CA3区神经元形态学影响,以及慢性砷中毒对成年大鼠脑的神经毒性。方法:取3～4月龄SD大鼠60只,随机分为正常对照组(自由饮用蒸馏水)、低剂量组(As2O3按4mg/l的比例用蒸馏水配制,自由饮用)和高剂量组(As2O3按100mg/l的比例用蒸馏水配制,自由饮用)。每组20只,各组均用普通饲料喂养,连续喂养4月后取大鼠海马组织,常规石蜡切片尼式染色(Nissl),并利用神经元特异性烯醇化酶(NSE)抗体进行SABC法免疫组化染色,观察大鼠海马CA3区神经元的形态学改变,进行细胞形态学计数及测量NSE阳性反应产物的平均光密度。结果:砷中毒组神经元数量减少,细胞形态不规则,可见大量坏死的神经细胞,NSE免疫组化染色阳性细胞减少(P0.01),阳性反应产物平均光密度降低(P0.01)。结论:慢性砷中毒可以导致大鼠海马CA3区神经元结构损伤。     

Nitric oxide and thyroid gland: modulation of cardiovascular function in autonomic-blocked anaesthetized rats

We have previously reported that acute administration of N(G)-nitro-l-arginine methyl ester (L-NAME) increases the mean arterial pressure (MAP) and heart rate (HR) in autonomic-blocked (CAB) anaesthetized rats. In the present study we examined whether thyroid and adrenal glands are involved in these pressor and chronotropic responses. Sprague-Dawley rats were studied after bilateral vagotomy and ganglionic blockade with hexamethonium (10 mg kg(-1)), and stabilization of MAP with infusion of phenylephrine (PE) (6 microg kg(-1) min(-1)). The rats were divided into groups: L, CAB; PE, CAB + PE bolus (6 microg kg(-1)); L-TX, thyroidectomy + CAB; L-AX, adrenalectomy + CAB; TX, only thyroidectomy; C, CAB. L, L-AX and L-TX groups received a bolus of l-NAME (7.5 mg kg(-1)). Triiodothyronine (T3), thyroxin (T4) and thyrotropin (TSH) levels were measured in L and L-TX rats before and after l-NAME administration. Reduced nicotamide adenine dinucleotide (NADPH) diaphorase activity was determined in heart and aorta of the TX group. The pressor response induced by l-NAME was similar in all groups. l-NAME-induced-tachycardia was associated with this rise in MAP. Adrenalectomy did not modify this chronotropic response, but it was attenuated by thyroidectomy. Thyroidectomy by itself decreased the circulating levels of T3 but it had no effect on the plasma levels of T4 and TSH. L and L-TX groups showed similar levels of circulating T4 and TSH, meanwhile the plasma level of T3 decreased in the L group. Nitric oxide synthase (NOS) activity in atria as well as in aorta was greater in the TX group compared with C. When autonomic influences are removed, the thyroid gland modulates intrinsic heart rate via a mechanism that involves, at least in part, the nitric oxide pathway.     

Effects of maternal state on the responsiveness to nest odors of hooded rats

Systemic antagonism of H1 or H2 receptors for histamine attenuated drinking elicited by SC 20 mg/kg histamine in adult male Sprague-Dawley rats. The H1 antagonist dexbrompheniramine (DXB; 0.5-16 mg/kg) and the H2 antagonist cimetidine (C; 0.5-100 mg/kg) each inhibited drinking elicited by histamine when given IP 10 min prior to SC histamine: The lowest doses to produce a statistically significant inhibition of drinking were 2 mg/kg DXB and 32 mg/kg C. While 1 mg/kg DXB alone or 16 mg/kg DXB plus 16 mg/kg C virtually abolished drinking elicited by histamine (1.25-20 mg/kg) in a dose-response study. In addition, such combined antagonism of H1 and H2 receptors failed to elicit drinking in the absence of exogenous histamine and failed to inhibit drinking elicited by deprivation from water for 7 or 24 hr. Because combined systemic antagonism of H1 and H2 receptors can specifically and completely inhibit drinking elicited by exogenous histamine, these findings provide a probe for a histaminergic component of drinking in the rat.     

Alternative complement pathway and factor B activities in rats with altered blood levels of thyroid hormone

Evaluating the activity of the complement system under conditions of altered thyroid hormone levels might help elucidate the role of complement in triggering autoimmune processes. Here, we investigated alternative pathway (AP) activity in male Wistar rats (180 ± 10 g) after altering their thyroid hormone levels by treatment with triiodothyronine (T3), propylthiouracil (PTU) or thyroidectomy. T3 and thyroxine (T4) levels were determined by chemiluminescence assays. Hemolytic assays were performed to evaluate the lytic activity of the AP. Factor B activity was evaluated using factor B-deficient serum. An anti-human factor B antibody was used to measure factor B levels in serum by radial immunodiffusion. T3 measurements in thyroidectomized animals or animals treated with PTU demonstrated a significant reduction in hormone levels compared to control. The results showed a reduction in AP lytic activity in rats treated with increasing amounts of T3 (1, 10, or 50 µg). Factor B activity was also decreased in the sera of hyperthyroid rats treated with 1 to 50 µg T3. Additionally, treating rats with 25 µg T3 significantly increased factor B levels in their sera (P < 0.01). In contrast, increased factor B concentration and activity (32%) were observed in hypothyroid rats. We conclude that alterations in thyroid hormone levels affect the activity of the AP and factor B, which may in turn affect the roles of AP and factor B in antibody production.     

The effect of lead and polychlorinated biphenyl exposure on rat natural killer cell cytotoxicity

Splenic natural killer (NK) cell cytotoxicity was assessed in rats chronically exposed to lead (Pb) as lead acetate in the drinking water or polychlorinated biphenyl (PCB) as Aroclor 1254 in the feed. Rats treated with cyclophosphamide were included as positive immunosuppressed controls. Weanling, male Sprague-Dawley rats exposed to 50 and 500 ppm PCB in the feed for ten weeks exhibited significantly suppressed (P less than 0.01) splenic NK activity. Cyclophosphamide injected i.p. six days prior to termination at a dose of 75 mg/kg also significantly inhibited splenic NK activity. NK cell activity was reduced, though not significantly, in spleen cells isolated from animals exposed to 10 and 1000 ppm Pb as Pb acetate in the drinking water for ten weeks. In vitro exposure of rat spleen cells to PCB at concentrations of 0.4 and 20.0 micrograms/ml similarly resulted in a significant depression of splenic NK cell activity. In addition, in vitro exposure to lead at the same concentrations resulted in suppressed NK cell cytotoxicity of rat splenocytes. These results indicate that two environmental contaminants have the ability to adversely affect NK cell cytotoxicity. The effects seen here with Pb and PCB on NK cells may in part explain the tumor inducing effect these chemicals are suspected of possessing via compromising the immune surveillance system.     

Developmental Exposure to 1.0 or 2.5 mg/kg of Dibutyltin Dichloride Does Not Impair Immune Function in Sprague-Dawley Rats

Organotins are used commercially as pesticides, antifouling agents, and stabilizers for polyvinyl chloride (PVC) pipe. Mono-and di-substituted butyltins, used in PVC pipe production, are of concern to the United States EPA, they leach from supply pipes into drinking water and are reported multisystem toxicants. We assessed immune function in Sprague-Dawley rats after developmental dibutyltin dichloride (DBTC) exposure. Pregnant rats were given drinking water containing 0, 10, or 25 mg/L of DBTC (final concentration) in 0.5% Alkamuls from gestational Day (GD) 6 through weaning of pups (37 days total). Approximate doses to dams: 1 and 2.5 mg DBTC/kg body weight (BW) during gestation, or 2.0 and 4.4 mg DBTC/kg BW while nursing. Litters were sexed, weighed, and culled to 4 males and 4 females per dam on postnatal Day (PND) 2. Beginning on PND3, litters of half of the dams per dose were gavaged with 0, 1.0, or 2.5 mg DBTC/kg BW 3X/week for 10 doses (maternal + direct treatment); remaining litters were exposed indirectly via lactation (maternal treatment). BW of litters exposed to 2.5 mg DBTC/kg BW was 10-20% lower (p < or = 0.05) relative to other groups from PND14 (males) or PND17 (females) through PND37. Delayed-type hypersensitivity (DTH), antibody synthesis, and natural killer (NK) cell activity were evaluated in immunologically mature offspring (N = 6/sex/group). DTH responses and antibody synthesis did not differ by dose, sex, or exposure. NK cell activity in the 10 mg DBTC/L maternal only group was greater in male offspring than in female. In female offspring from the maternal + direct group, cytotoxicity increased by dose at the 25:1 effector:target cell ratio. Our data suggest that developmental immunotoxicity from DBTC-tainted drinking water is unlikely as the concentrations we used were several orders of magnitude higher than concentrations expected to leach from PVC pipes.     

Suppression of drinking by exposure to a high-strength static magnetic field

High-strength static magnetic fields of 7 T and above have been shown to have both immediate and delayed effects on rodents, such as the induction of locomotor circling and the acquisition of conditioned taste aversions. In this study, the acute effects of magnet field exposure on drinking were examined. Exposure to a 14.1-T magnetic field for as little as 5 min significantly decreased the amount of a glucose and saccharin solution (G+S) consumed by water-deprived rats over 10 min. The decreased intake could be accounted for largely, but not entirely, by an increase in the latency of magnet-exposed rats to initiate drinking. When intake was measured for 10-60 min after the initiation of drinking, thus controlling for increased latency, magnet-exposed rats still consumed less G+S than sham-exposed rats. The increased latency was not due simply to an inability of magnet-exposed rats to reach the elevated sipper tube of the G+S bottle, providing rats with long tubes that could be reached without raising their heads normalized intake but latency was still increased. The increased latency and decreased intake appeared to be secondary to somatic effects of magnet exposure, however, because during intraoral infusions magnet-exposed rats consumed the same amount of G+S with the same latency to reject as sham-exposed rats. The suppression of drinking by magnetic field exposure is consistent with the acute effects of other aversive stimuli, such as whole-body rotation, on short-term ingestion. These results add to the evidence that high-static strength magnetic fields can have behavioral effects on rodents.     

A small amount of tetrachloroethylene ingestion from drinking water accelerates antigen-stimulated allergic responses

Previously, we observed that tetrachloroethylene (perchloroethylene, PCE) increased histamine release and inflammatory mediator production from antigen-stimulated mast cells. In this study, we examined the enhancing effect of low concentrations of PCE in drinking water on antigen-stimulated allergic responses. After exposure of Wistar rats to PCE in drinking water for 2 or 4 weeks, we performed a passive cutaneous anaphylaxis (PCA) reaction. PCE exposure for 4 weeks enhanced PCA reaction in a dose-dependent manner. In pathological studies, PCE exposure for 2 weeks exacerbated inflammation characterized by infiltration of lymphocytes and accumulation of mast cells around the vessel. Non-purified mast cells (NPMCs) from rats treated with 1mg/L PCE in drinking water for 2 weeks increased antigen-stimulated histamine release. Furthermore, the leukocytes of rats treated with PCE in drinking water for 4 weeks showed increased interleukin (IL)-4 expression. The mechanism of enhancing the PCA reaction is assumed to be that PCE increases IL-4 production and PCE causes T helper (Th) 1/Th2-type helper T-cell imbalance and increases histamine release from excessively accumulated mast cells. The results suggest that the intake of PCE in drinking water, even at a low concentration, leads to the initiation and acceleration of allergic diseases.     

Characteristics of inflammatory cells in spontaneous autoimmune thyroiditis of NOD.H-2h4 mice

Thyroid lesions develop in most NOD.H-2h4 mice 6 weeks after they are given 0.05% NaI in drinking water. B cells are required for spontaneous autoimmune thyroiditis (SAT) development, and anti-thyroglobulin autoantibody levels correlate with SAT severity. Immunohistochemical staining of thyroids obtained 2-10 weeks after administration of NaI water suggested that CD4+ T cells initially infiltrated the thryoid, followed by CD8+ T cells and B cells. Intrathyroidal CD4+ T cells are more numerous than CD8+ T cells. CD4+ T cells and B cells form aggregates in the thyroid, while CD8+ T cells are scattered throughout the thyroid. Intrathyroidal germinal centre-like structures could be observed in thyroid lesions with 2-3+ SAT and intrathyroidal B cells co-expressed OX40L. By RT-PCR, intrathyroidal expression of OX40L, OX40, CD40L, IL-2R, CTLA-4 and Igbeta mRNA correlated closely with the SAT severity score. These molecules were not expressed in normal thyroids. In the spleen, OX40L-positive cells were detected at 2 weeks and increased 4-6 weeks after NaI water. OX40, OX40L, CD40L, IL-2R and B7-1 as well as IFN-gamma and IL-4 mRNA were minimally expressed in normal spleens, usually began to be expressed at 2 weeks and increased to maximal level 4-8 weeks after NaI water. These results suggest that in NOD.H-2h4 mice, the OX40L, OX40, CD40L and B7 molecules, which increase in the spleen and thyroid of these mice after receiving NaI water, may play a role in SAT development, implying that one or more of these molecules might be good targets for the prevention or treatment of SAT.     

Histaminergic mechanism for drinking elicited by insulin in the rat

Sprague-Dawley male albino rats showed in a dose-response study a maximal drinking response to a 5 U/kg dose of SC insulin in a 2-hr test. Drinking elicited by 5 U/kg insulin was reduced to baseline (i.e., no insulin) level by combined antagonism of H1 and H2 receptors for histamine using IP 1 mg/kg dexbrompheniramine plus 16 mg/kg cimetidine. Antagonism of histamine receptors in this fashion was specific for drinking elicited by histamine because such antagonism reduced to baseline level drinking elicited by 2.5 mg/kg SC histamine, but failed to inhibit drinking after 8- or 24-hr water deprivation or drinking after 0.63 mg/kg SC serotonin (5-HT). These results demonstrate a histaminergic mechanism for drinking elicited by exogenous insulin which is consistent with the published report that exogenous insulin can release gastric mucosal histamine in the rat. Moreover, because eating is known to elicit the release of endogenous insulin, the results reported here suggest a working hypothesis that endogenous insulin is a component for drinking around mealtime in the rat.     

Sequential analysis of development of invasive thyroid follicular cell carcinomas in inflamed capsular regions of rats treated with sulfadimethoxine after N-bis(2-hydroxypropyl)nitrosamine-initiation

A 2-stage thyroid follicular carcinogenesis model in rats initiated with N-bis(2-hydroxypropyl)nitrosamine (DHPN) is widely used to detect modifying effects of chemicals on thyroid carcinogenesis. A number of goitrogens are known to strongly promote carcinogenesis, and the carcinomas often originate adjacent to the thyroid capsule and show invasive growth into the capsule or adjacent tissues. To clarify mechanisms of progression to invasive carcinomas, we sequentially evaluated histopathological and immunohistochemical characteristics of thyroids in male F344 rats treated with sulfadimethoxine (SDM, 0.1% in drinking water) for 0-10 weeks beginning 1 week after DHPN initiation (2800 mg/kg body weight, single s.c. injection). In DHPN-SDM-treated rats, multiple focal hyperplasias and adenomas developed in thyroid follicular parenchyma at weeks 4 to 6. Apart from the proliferative lesions, capsular thickening with inflammatory cell infiltration, mainly consisting of macrophages, and migration of follicular epithelium into the capsule were also observed. Focal hyperplasias/adenomas adjacent to the capsule progressively developed to invasive carcinomas at weeks 6 to 10. In thyroid parenchyma, malignant lesions were seldom observed. With SDM-treatment alone, although no neoplastic lesions were observed, capsular thickening with inflammation and epithelial migration resulted in intracapsular residual follicles. Intracapsular residual follicular cells as well as invasive and intrathyroidal carcinoma cells generally showed increased cell proliferative activity, coincidental with cytoplasmic/nuclear positivity for beta-catenin. These results suggested that beta-catenin activation related to capsular inflammation may play a role in development of invasive carcinomas but is insufficient for tumor formation by itself. Whether this is associated with mutations in the beta-catenin gene remains to be clarified.     

Cholecystokinin blockade alters the systemic immune response in rats with acute pancreatitis

Acute pancreatitis (AP) is characterized by initial pancreatic injury resulting from the activation of digestive enzymes and, later, widespread inflammation to distant organs. The aim of this study was to study whether the time-course of inflammatory events during AP induced by bile-pancreatic duct obstruction (BPDO) varies after lowering the acinar enzyme content by L364,718 (0.1 mg/kg/day) administration over 7 days before inducing AP. Flow cytometric immunophenotyping was used to analyse the following at different AP stages: distribution of major circulating leucocyte subsets, activation state of circulating neutrophils and monocytes as reflected by CD11b expression and tumour necrosis factor-alpha (TNF-alpha) production and the contribution of T-cell-derived pro-(TNF-alpha) and anti-(IL-10) inflammatory mediators. TNF-alpha plasma levels and neutrophil infiltration in pancreas and lung were also measured. At early BPDO times, L364,718 treatment partially inhibited leukocytosis and increase in peripheral blood neutrophils and monocytes as well as TNF-alpha expression by monocytes. However, from 6 h onwards after BPDO, L364,718 treatment was unable to prevent either pancreatic and lung neutrophil infiltration or the release of TNF-alpha from activated monocytes. By its action on circulating lymphocytes, L364,718 treatment enhanced the severity of the inflammatory response induced by BPDO. Peripheral blood lymphocytes were recruited from earlier BPDO times, and 12 h after BPDO, T cells displayed a significantly higher reserve of TNF-alpha able to be released under stimulation but lower functional reserve of interleukin-10 (IL-10) than observed in untreated rats. It is concluded that lowering the acinar enzyme content through L364,718 treatment prevents earlier systemic immune events in BPDO-induced AP. However, at the point of maximal injury, the inflammatory response became pronounced, largely due to the role played by activated T lymphocytes.     

急性胰腺炎大鼠腹水白细胞cPKC-α水平的变化及汉防己甲素等的影响

目的:观察急性胰腺炎(AP)大鼠腹水白细胞传统型蛋白激酶C-α(cPKC-α)的水平变化及汉防己甲素(Tet)等药物的影响,并对其机制进行探讨。方法:取56只SD大鼠随机分为4组:假手术组(SO+NS组),AP对照组(AP+NS组),AP+阿司匹林(ASP)组,AP+Tet组,采用牛磺脱氧胆酸钠复制AP模型,模型制成后10min,AP+Tet组腹腔注入Tet(80mg/kg·BW);AP+ASP组经食管灌入ASP混悬液(125mg/kg·BW)。各组动物在相应处理后1h、5h,提取腹水白细胞,制备白细胞裂解液,用免疫印迹(Westernblot)和化学发光法检测其cPKC-α水平;同时分别取动物血浆测定淀粉酶(AMY)活性;取5h大鼠胰腺组织作病理检查。结果:ASP和Tet组胰腺病理损伤轻于AP+NS组,血浆AMY活性低于AP+NS组,腹水白细胞cPKC-α水平高于AP+NS组(P＜0.05或P＜0.01)。结论:ASP、Tet可增加AP大鼠腹水白细胞cPKC-α的水平;ASP、Tet可明显减轻AP时胰腺组织及功能损伤。     

Carcinogenicity of chloral hydrate administered in drinking water to the male F344/N rat and male B6C3F1 mouse

Male B6C3F1 mice and male F344/N rats were exposed to chloral hydrate (chloral) in the drinking water for 2 years. Rats: Measured chloral hydrate drinking water concentrations for the study were 0.12 g/L, 0.58 g/L, and 2.51 g/L chloral hydrate that yielded time-weighted mean daily doses (MDDs) of 7.4, 37.4, and 162.6 mg/kg per day. Water consumptions, survival, body weights, and organ weights were not altered in any of the chloral hydrate treatments. Life-time exposures to chloral hydrate failed to increase the prevalence (percentage of animals with a tumor) or the multiplicity (tumors/animal) of hepatocellular neoplasia. Chloral hydrate did not increase the prevalence of neoplasia at any other organ site. Mice: Measured chloral hydrate drinking water concentrations for the study were 0.12 g/L, 0.58 g/L, and 1.28 g/L that gave MDDs of 13.5, 65.0, and 146.6 mg/kg per day. Water consumptions, survival, body and organ weights, were not altered from the control values by any of the chloral hydrate treatments. Enhanced neoplasia was observed only in the liver. Prevalence and multiplicity of hepatocellular carcinoma (HC) were increased only for the high-dose group (84.4%; 0.72 HC/animal; p < or = 0.05). Values of 54.3%; 0.72 HC/animal and 59%; 1.03 HC/animal were observed for the 13.5- and 65.0-mg/kg per day treatment groups. Prevalence and multiplicity for the control group were 54.8%; 0.74 HC/animal. Hepatoadenoma (HA) prevalence and multiplicity were significantly increased (p < or = 0.05) at all chloral hydrate concentrations: 43.5%; 0.65 HA/animal, 51.3%; 0.95 HA/animal and 50%; 0.72 HA/animal at 13.5, 65.0, and 146.6 mg/kg per day chloral hydrate compared to 21.4%; 0.21 HA/animal in the untreated group. Altered foci of cells were evident in all doses tested in the mouse, but no significant differences were observed over the control values. Hepatocellular necrosis was minimal and did not exceed that seen in untreated rats and mice. Chloral hydrate exposure did not alter serum chemistry and hepatocyte proliferation in rats and mice or increase hepatic palmitoyl CoA oxidase in mice at any of the time periods monitored. It was concluded that chloral hydrate was carcinogenic (hepatocellular neoplasia) in the male mouse, but not in the rat, following a lifetime exposure in the drinking water. Based upon the increased HA and combined tumors at all chloral hydrate doses tested, a no observed adverse effect level was not determined.     

Immune responses in sprague-dawley rats exposed to dibutyltin dichloride in drinking water as adults

Organotins are used commercially as agricultural pesticides, antifouling agents, and stabilizers for polyvinyl chloride (PVC) pipe. Mono- and di-substituted methyl and butyltins, used in PVC pipe production, are of concern as they leach from supply pipes into drinking water and have been reported to cause multisystem toxicity, including immunotoxicity. As part of an ongoing study to evaluate immunotoxic effects of organotins, we assessed immune function in adult Sprague-Dawley (CD) rats after exposure to dibutyltin dichloride (DBTC). Individually-housed adult male and female CD rats were given drinking water containing 0, 10, or 25 mg DBTC/L (final concentration) in 0.5% Alkamuls for 28 days. Water bottles were changed and water consumption was monitored twice weekly and body weights (BW) were recorded weekly. Delayed-type hypersensitivity (DTH), primary and secondary antibody responses to sheep red blood cells, and natural killer (NK) cell activity were evaluated in separate groups of treated and control animals on day 29 of exposure. Water consumption was significantly decreased in both sexes at 25 mg DBTC/L. BW, immune organ weights, the DTH response, and NK cell activity did not vary by dose. Different results for antibody responses in male rats were obtained in two experimental replicates. In the first replicate, IgG was elevated at the highest dose whereas in the second replicate, IgM was suppressed. However, as these effects occurred at the high dose of 25 mg DBTC/L, which is a concentration a million times higher than levels of DBTC reported in drinking water, our data suggest that DBTC is unlikely to cause immunotoxicity at concentrations found in drinking water supplies.     

Histamine plays no part in schedule-induced polydipsia in the rat

Twelve Sprague-Dawley male albino rats were tested with or without combined antagonism of peripheral H1 (using 2 mg/kg dexbrompheniramine IP) and H2 (using 32 mg/kg cimetidine IP) receptors for histamine prior to (a) drinking after 2.5 mg/kg histamine SC, (b) drinking after 24-hr water deprivation, and (c) drinking during the acquisition and maintenance of schedule-induced polydipsia (SIP) with a 45 mg Noyes pellet delivered every 90 sec. Such antagonism of histamine receptors abolished drinking elicited by exogenous histamine without inhibiting drinking after water deprivation. Moreover, histaminergic antagonism failed to prevent the acquisition and maintenance of SIP and failed to alter the distribution of contacts with the drinking spout during interpellet intervals. These findings demonstrate no role for endogenous systemic histamine in SIP.