Passage of hybridomas in male BALB/c mice

For cultivating hybridomas in the ascitic form there are usually used female mice BALB/c and not male ones. Efficiency of production of monoclonal antibodies with cultivation of the hybridomas in male and female mice BALB/c was studied comparatively. The animals were stimulated to form ascite by administration of the incomplete Freund's adjuvant or 3 per cent peptone with petrolatum oil. Some parameters of the ascite formation were studied: viability of the hybridoma cells, ascitic fluid formation period and volume, hybridoma cell concentration and titers of monoclonal antibodies in the ascitic fluid. In regard to all the parameters studied the male animals were not inferior to the female ones and in case of one of the hybridomas even surpassed them twofold by the volume of the ascitic fluid formed. This is evident of possible using male mice for mass cultivation of hybridoma cells with a purpose of obtaining preparative amounts of monoclonal antibodies in production of immunodiagnostic agents on their basis.     

Subcutaneously administrated genistein and daidzein decrease serum cholesterol and increase triglyceride levels in male middle-aged rats

Nutritional supplements containing soybean phytoestrogens, the isoflavones genistein (G) and daidzein (D), are increasingly used as alternative therapy for osteoporosis, cancer, and cardiovascular and other diseases with a frequency that increases with advancing age. In this study we examined the effects of subcutaneous administration of either G or D on serum lipid levels in orchidectomized (Orx) and intact (IA) middle-aged male rats, which are experimental models of andropause. Sixteen-month-old Wistar rats were treated with 10 mg/kg and 30 mg/kg of either G or D. The control groups received testosterone, estradiol, or vehicle for 3 weeks, after which the total serum cholesterol (TC), low-density lipoprotein (LDL-C), high-density lipoprotein (HDL-C), and total triglycerides (TT) were measured. Compared with the matching vehicle-treated controls, the higher doses of G and D and testosterone treatment significantly (P < 0.05) lowered the TC and lipoprotein cholesterol levels. The greatest effect was observed regarding LDL-C in both Orx and IA males after G and D treatments, in which LDL-C decreased by more than 30%. The lower isoflavone doses induced a significant cholesterol-lowering effect (P < 0.05) only in the Orx group. Like the estradiol treatment, the higher doses of G and D increased the TT levels in both rat models by more than 50% (P < 0.05). The lower doses of isoflavones increased TT only in the Orx group. In male middle-aged rats, injections of higher doses of G and D decreased the serum cholesterol levels, as did testosterone injection, and brought about an increase in serum triglycerides similar to that observed after estradiol treatment.     

Effects of hCG on serum levels of testosterone, dihydrotestosterone and androstenedione in male mice

The concentrations of testosterone (T), 5alpha-dihydrotestosterone (5alpha-DHT) and delta4-androstenedione (delta4 A) in the circulating blood were increased by a single subcutaneous injection of 10 IU of hCG in the mouse. The response of androgen synthesis to hCG stimulation was rapid and persisted over a period of 96 h. The increased supply of T to the peripheral organ(s) under the stimulation of hCG might contribute to an increase in circulating 5alpha-DHT. However, the changes in T/5alpha-DHT ratio implies that the stimulation of T and 5alpha-DHT production by hCG were not of the same degree.     

Effects of orally administered retinol on natural killer cell activity in wild type BALB/c and congenitally athymic BALB/c mice

Summary Retinoids have been shown to inhibit the growth and development of neoplastic cells in many systems. One mechanism of action may be through activation of the immune system, specifically natural killer (NK) cell activity. The effect of retinol on NK cell cytotoxicity was examined in three groups of mice: BALB/c (wild-type), BALB/c nu/nu (athymic), and BALB/c nu/nu previously injected with human tumor cells. In untreated mice, NK activity was highest in athymic mice without tumors and lowest in wild-type mice, although serum and liver retinol concentrations were identical in all three groups. In mice fed graded, nontoxic doses of retinol daily for 3 weeks, serum retinol levels in all three groups exhibited a sharp peak and decline following daily bolus retinol administration. Retinol stores in the livers showed a dose-dependent increase in all treated animals. However, NK cell activity, differed for each group. Athymic mice without tumors exhibited no change in NK activity as a result of retinol treatment. Athymic mice with tumors had NK levels that tended to increase with increasing retinol doses, but these changes were not statistically significant. Wild-type mice, on the other hand, demonstrated significantly higher NK levels after treatment with retinol doses of 300 and 600 g/day. In subsequent time course experiments, there was a peak in NK activity 1 h following bolus retinol administration similar to the peak seen in serum retinol concentrations, suggesting either an acute activation or recruitment of cytotoxic cells. Retinol thus appears to increase NK activity in wild-type BALB/c mice, and this activity may be an important component of its antineoplastic activity.This investigation was supported by Biomedical Research Support Grant RRO 5424, the Veterans Administration, and PHS Grant number CA 33589-01A2, awarded by the National Cancer Institute, DHHSThis work was done in partial fulfillment of a Ph. D. thesis by L. Fraker in the Department of Pathology, Vanderbilt University School of Medicine     

Effects of carcinogenic and non-carcinogenic chemicals on plasma esterases in BALB/c mice

Esterase profiles of plasma from female BALB/c mice treated with a variety of carcinogenic and weakly- or non-carcinogenic chemicals were analyzed. Mice treated with the potent carcinogens diethylnitrosamine, dinitrosopiperazine, dipropylnitrosamine, dimethylhydrazine, urethane, and dimethyldinitrosopiperazine had similarly altered plasma esterase profiles after 7 days' exposure to the chemicals. The alterations consisted of increased activity in 4 esterase bands. The increased activity persisted in some of the bands after cessation of carcinogen exposure. Exposure to high concentrations of the weakly- or non-carcinogenic compounds nitrosohydroxyproline, nitrosomethoxymethylamine, 1-nitroso-4 methylpiperazine, nitroso-2,6 dimethylpiperidine, and ethyl methanesulfonate caused no obvious plasma esterase alterations. Ingestion of carbon tetrachloride resulted in increased activity in one esterase band with concomitant decrease in a second band. Analysis of serum from test mice for levels of serum glutamic oxaloacetic transaminase, alkaline phosphatase, lactate dehydrogenase-lactate substrate, and D-gamma-glutamyl transpeptidase did not differentiate between mice treated with selected carcinogens and those treated with non-carcinogens and/or carbon tetrachloride.     

Elaiophylin reduces body weight and lowers glucose levels in obese mice by activating AMPK

Obesity is an epidemic affecting 13% of the global population and increasing the risk of many chronic diseases. However, only several drugs are licensed for pharmacological intervention for the treatment of obesity. As a master regulator of metabolism, the therapeutic potential of AMPK is widely recognized and aggressively pursued for the treatment of metabolic diseases. We found that elaiophylin (Ela) rapidly activates AMPK in a panel of cancer-cell lines, as well as primary hepatocytes and adipocytes. Meanwhile, Ela inhibits the mTORC1 complex, turning on catabolism and turning off anabolism together with AMPK. In vitro and in vivo studies showed that Ela does not activate AMPK directly, instead, it increases cellular AMP/ATP and ADP/ATP ratios, leading to AMPK phosphorylation in a LKB1-dependent manner. AMPK activation induced by Ela caused changes in diverse metabolic genes, thereby promoting glucose consumption and fatty acid oxidation. Importantly, Ela activates AMPK in mouse liver and adipose tissue. As a consequence, it reduces body weight and blood glucose levels and improves glucose and insulin tolerance in both ob/ob and high-fat diet-induced obese mouse models. Our study has identified a novel AMPK activator as a candidate drug for the treatment of obesity and its associated chronic diseases.Subject terms: Cell biology, Drug discovery     

Effects of estrogen on food intake, body weight, and temperature of male and female obese mice

Estradiol benzoate (EB) treatment of male and female C57BL/6J ob/ob mice for 32 days led to decreased body weight (20%), percentage body fat (8%) and carcass protein content (12%) when compared with non-EB-treated obese control mice. Estradiol reduced the caloric intakes of both genders by 25-35%, but did not affect body temperature regulation. Circulating glucose and insulin concentrations were also lowered by estrogens, although hyperinsulinemia persisted. Since post-treatment body weight changes correlated with daily food intakes (r = 0.81) rather than to rectal temperatures (r = -0.19), it appears that hypophagia provided a greater contribution to the estrogen-mediated reductions of growth and carcass fat than did altered energy expenditure for thermoregulation. While these data show that EB treatment does reduce the severity of some metabolic disturbances in a genetic model of type II diabetes, long-term estrogens do not appear to offer substantial advantages in the treatment of obesity or diabetes when compared with the effects of caloric restriction alone.     

Sub-chronic administration of stable GIP analog in mice decreases serum LPL activity and body weight

GIP receptor knockout mice were shown to be protected from the development of obesity on a high fat diet, suggesting a role of GIP in the development of obesity. In our study we aimed to test the hypothesis if excess of GIP could accelerate development of obesity and to identify GIP gene targets in adipose tissue. Therefore, mice were kept on a chow or a high fat diet and during the last 2 weeks D-Ala²-GIP or PBS injections were performed. Afterwards, serum LPL activity and several biochemical parameters (TG, FFA, cholesterol, glucose, insulin, resistin, IL-6, IL-1β, TNFα, GIP) were measured. Fat tissue was isolated and QPCR was performed for a set of genes involved in energy metabolism and inflammation. A DNA-microarray was used to identify GIP gene targets in adipose tissue of the chow diet group. We found that the D-Ala²-GIP injections caused a significant decrease in both body weight and LPL activity compared to controls. Serum biochemical parameters were not affected by D-Ala²-GIP, with an exception for resistin and insulin. The set of inflammatory genes were significantly decreased in adipose tissue in the D-Ala²-GIP injected animals on a chow diet. A DNA-microarray revealed that APO-genes and CYP-genes were affected by D-Ala²-GIP treatment in adipose tissue. These results suggest that the body weight-reducing effect of D-Ala²-GIP may be explained by lower LPL activity and insulin serum level. Moreover, the identified GIP candidate gene targets in adipose tissue link GIP action to lipid metabolism exerted by APO and CYP genes.     

Effects of nicotine on body weight, food consumption and body composition in male rats

The present study examined the effects of nicotine administration and cessation on body weight, food consumption, and body composition in rats. Administration of nicotine was associated with attenuated body weight gains and cessation was associated with accelerated body weight gains. Changes in fat composition paralleled changes in body weight, whereas changes in body water and protein did not. These results suggest that nicotine administration decreases body weight through its effects on fat stores in the body. After cessation of nicotine, body fat rapidly approached levels of control animals.     

Effects of hyperthermia and photodynamic therapy on BALB/c mice bearing subcutaneous tumors

The therapeutic effects of photodynamic therapy and hyperthermia on mice bearing subcutaneous tumors were investigated. Ehrlich ascites tumor cells (1 x 10(7)) were implanted subcutaneously into the femoral area of BALB/c mice. A total of 134 tumor-bearing mice were treated with photodynamic therapy, i.e., administration of laser irradiation (514.5 nm, 112.5 mW/cm2 for 11.12 min with a total energy 75 J/cm2) after injection (i.p.) of hematoporphyrin derivative (HPD, 7.5 and 10.0 mg/kg body weight) and/or hyperthermia (by electric heating needles to 44 and 45 degrees C for 30 min) once a day for three successive days. The results revealed that the therapeutic effects of the combination of photodynamic therapy and hyperthermia were improved when compared with photodynamic therapy or hyperthermia alone. A combination of photodynamic therapy (10.0 mg HPD/kg body weight and 75 J/cm2 of total laser irradiation energy) and hyperthermia (44 degrees C for 30 min) had the best therapeutic effect, indicating that the mortality rate within 120 days (MR120) was 12.5% and the mean survival time (MST120) was 113.8 days.     

Influence of experimental diets on cholesterol and triglyceride levels of rabbit blood serum lipoproteins

Groups of rabbits were fed for six weeks various diets: standard died + ethanol, high-cholesterol diet and a high-cholesterol + ethanol one. During the next six weeks every diet was supplemented with a fresh vegetable (carrot). Cholesterol and triglycerides were determined in the whole serum and in lipoprotein fractions. In rabbits fed standard diet ethanol caused a moderate elevation of VLDL cholesterol and triglyceride and LDL cholesterol levels. In animals on high-cholesterol diet cholesterol and triglyceride concentrations in these fractions were very high. Simultaneous consumption of large amounts of cholesterol and of ethanol resulted in a greater rise of cholesterol concentration in the whole serum and in VLDL and LDL fraction than did high-cholesterol diet alone. Addition of carrot caused a pronounced reduction of serum cholesterol concentration in animals fed all kinds of diets. The reduction concerned mainly VLDL.     

Effects of galanin-like peptide (GALP) on locomotion, reproduction, and body weight in female and male mice

Galanin-like peptide (GALP) has been implicated in the neuroendocrine regulation of both feeding and reproduction. In male rodents and primates, intracerebroventricular (icv) infusions of GALP stimulate luteinizing hormone (LH) release, induce Fos expression in brain areas implicated in feeding and reproduction, and affect food intake and body weight in rodents. In gonad-intact and castrated male rats, icv administration of GALP also stimulates male sexual behavior. While the effects of GALP on male physiology and behavior are well documented, no studies have addressed such a role of GALP in females. We tested the effects of icv GALP infusions on LH release, locomotor activity, motor control, and body weight regulation in adult ovariectomized female mice hormonally primed with estradiol benzoate and progesterone. In addition, sexually-experienced male and female mice were treated with GALP and tested for sexual behavior. In females, GALP reduced open-field locomotor activity, the ability to maintain grip on an accelerating rotarod, and 24-h body weight in a dose-dependent manner. GALP also increased LH secretion in female mice, an effect that was blocked by pre-treatment with Antide, a gonadotropin-releasing hormone (GnRH) type-1 receptor antagonist. GALP infusions slightly decreased the occurrence of lordosis behavior in female mice and significantly increased the latencies with which females displayed receptivity. Unlike previous reports in male rats, GALP inhibited male sexual behavior in mice. Our data indicate that in female mice, GALP stimulates LH release via GnRH, and decreases body weight, motor control, and locomotor activity via GnRH-independent pathways. Furthermore, our sexual behavior and locomotor findings suggest species-specific differences in the mechanism and/or location of GALP action in the brains of rats and mice.     

Plasma triglyceride levels correlate with body weight,age and gender influence body weight changes in Spermophilus lateralis during prehibernation and hibernation

Our experimental groups consisted of a total 16 male (M) and 15 female (F), juvenile and adult Spermophilus lateralis. This study evaluated weight gain rate (WGR) and mass-specific rate of gain (MSRG) during prehibernation, and weight loss rate (WLR), mass-specific rate of loss (MSRL) plus changes in the levels of plasma triglyceride (TG) during hibernation in these squirrels (SQ). Between January and March, the plasma samples were obtained from 26 SQ. Plasma TG was determined enzymatically, and it averaged 189 mg% with no significant difference in gender. Correlations were observed between plasma TG and sacrificed body weight (r = 0.441, p < 0.05), and between plasma TG and MSRL (r = 0.409, p < 0.05), although plasma TG was not correlated with WLR during hibernation. Juvenile female SQ had a higher WGR than adult female SQ, but no significant difference was found between juvenile and adult males. Both juvenile males and females showed a significantly higher MSRG than adult male and female SQ. Conversely, adult male SQ had a higher WLR and MSRL than juvenile male SQ, but no significant difference was observed between adult and juvenile females. Moreover, juvenile male SQ showed a significantly lower WLR but not MSRL than juvenile females. For these animals, our results suggest that the age factor influences on WGR and MSRG during prehibernation, and also influences on WLR and MSRL during torpor.     

Reproductive tract and pancreatic norepinephrine levels in pre- and overt-diabetic C57BL/KsJ mice: relationship to body weight, blood glucose, serum insulin, and reproductive dysfunction

Diabetes-associated changes in tissue norepinephrine (NE) levels were determined in pre- and overt-diabetic C57BL/KsJ (db/db) mice relative to age-matched controls (+/?). At 4 weeks of age, both pre- and overt-diabetic mice exhibited obesity and hyperinsulinemia relative to controls. Prediabetic mice demonstrated normoglycemia and tissue (i.e., pancreatic, ovarian, and uterine) NE levels which were comparable to control levels. However, the hyperglycemic condition that characterized the overt-diabetic mice was found to be associated with significant elevations in tissue NE levels relative to both control and prediabetic values. These data demonstrate that the Type II diabetes-obesity syndrome in this mutant murine model is accompanied by a dramatic imbalance in the adrenergic influence on reproductive tissue glucose homeostasis. The hyperglycemic component of the diabetes-obesity syndrome is temporally linked to the noradrenergic disturbances in this model, which occur independently of insulin- or obesity-related changes that also accompany the expression of the genomic mutation and reproductive tract involution.     

Qualitative alterations in plasma esterases in BALB/c mice following the administration of diethylnitrosamine.

Selected non-specific plasma esterases in female BALB/c mice, separated by polyacrylamide gel electrophoresis, were qualitatively altered following treatment of the mice with diethylnitrosamine (DEN). These alterations include a decreased preference for naphthyl butyrate as a substrate relative to naphthyl acetate; decreased sensitivity to enhancement by divalent cations such as Ca2+, Mg2+, and Mn2; and an increased sensitivity to inhibition by eserine. All esterase species affected were also quantitatively altered and some were testosterone-dependent.     

Effects of oral administration of nicotine on organ weight, serum testosterone level and testicular histology in adult male rats

This study investigated the effects of oral administration of nicotine on body and reproductive organ weight, serum testosterone level and testicular histology in adult male rats. Forty male rats divided into five groups and treated for a period of 30 days with 0.5mg/kg (low dose) and 1.0 mg/kg (high dose) body weight of nicotine while the control rats received 0.2 ml/kg normal saline. The fourth and fifth groups were gavaged with 0.5mg/kg and 1.0mg/kg body weight of nicotine but were left untreated for another 30 days. These groups served as the recovery groups.  At the end of each experimental period, the animals were scarified and their reproductive organs were removed and weighed immediately. There was no significant change in the body weight. There was a significant decrease (p <0.05) in the testicular and epididymal weight of rats for both treatments while the decrease in the seminal vesicle weight for both treatment groups was not significant. The prostate weight was not significantly increased in both groups. The recovery groups showed appreciable recovery in their organ weight. Serum level of testosterone of both groups was significantly decreased in a dose dependent manner when compared with those of the control rats. The histological section showed testicular degeneration and disorganization in the cytoarchitecture, as the observed changes were pronounced in the high dose group than the low dose group. However, there were both regeneration of the germinal epithelium and restructuring of the interstitum towards normal in the recovery groups. No lesion was observed in the epididymis of the rats. The results suggest that nicotine has deleterious effect on the male reproductive organ of albino rats ameliorated by nicotine cessation.     

当归挥发油对哮喘BALB/c小鼠的平喘作用及对Th17免疫活性的影响

目的：探讨当归挥发油对哮喘BALB/c小鼠的平喘作用及对IL-17A、RORγt等Th17细胞活性因子的影响。方法：取雌性小鼠随机分为7组（n=12）：正常对照组、模型对照组、地塞米松组、当归挥发油高中低剂量组及当归油中+地米组,通过注射卵清白蛋白（OVA）致敏、吸入OVA激发的方法来复制哮喘动物模型,在当归挥发油的防治下,考察当归挥发油对哮喘行为学、肺功能、肺组织病理学、血清IL-17A及肺组织RORγt表达的影响。结果：60、120、240 mg/kg当归挥发油可维持哮喘小鼠体重的正常增长,改善哮喘行为学、肺功能及肺组织病理学变化,抑制IL-17A与RORγt的过度表达（P<0.05, 0.01）。同时,当归挥发油与地塞米松配伍后对维持体重的正常增长、缓解IL-17A与RORγt的过度表达具有一定的协同作用。结论：当归挥发油具有明显的防治哮喘作用,抑制IL-17、RORγt过度表达而抑制Th17细胞免疫活性是其作用机制之一;而当归挥发油与糖皮质激素配伍后有一定的协同作用。     

Staphylococcal enterotoxin B toxicity in BALB/c mice: Effect on T-cells, plasma cytokine levels and biochemical markers

Abstract Groups of BALB/c mice were treated with a sub-lethal dose (60 μg) of staphylococcal enterotoxin B (SEB) intraperitoneally and were sacrificed at 2, 5, 8, or 10 h post-injection. Organ, blood plasma and lymph node samples from these mice were analyzed. Plasma levels of urea, creatinine and alanine aminotransferase were significantly raised above normal by 5 h post-injection. However, alkaline phosphatase levels showed an erratic increase after toxin administration and, after administration of 10–40 μg SEB per mouse, were consistently at least 30% below normal levels at 24 h post-injection. Weight change was also monitored but found to be inconsistent. Lung, spleen and kidney samples appeared normal on histopathological examination, but liver samples showed minor polymorph infiltration and congestion. TNF-α, and IL-1 α levels in the plasma were raised by 8 h to picogram levels per ml of plasma, whereas IFN-γ and IL-2 were raised by 2 h to nanogram levels per ml of plasma. Lymph node cells taken from mice treated with toxin were given a secondary stimulation with toxin in vitro. Although the response of the cells was lower than normal on assay at four days, a time response curve showed a peak in cell responsiveness to secondary stimulation with toxin at three days. These data indicate that biochemical markers and cytokine levels are affected by the administration of SEB to mice and may be used as indicators of toxicity.