High-density lipoprotein associated with secondary vitellogenesis in the hemolymph of the crayfish Cherax quadricarinatus.

The high-density lipoproteins LPI and LPII were isolated from the hemolymph of the crayfish Cherax quadricarinatus by gradient ultracentrifugation and high-performance liquid chromatography (HPLC). Both lipoproteins contained a carotenoid moiety. LPI is comprised of a single polypeptide with an approximate molecular mass of 96 kDa. LPII was composed of two similar native components, LPIIa and LPIIb, both having polypeptides of 80 and 177 kDa. Both under natural conditions and after endocrine manipulations, LPI was present in males and in females, regardless of the female reproductive stage. LPII was present only in secondary-vitellogenic females, but not during the winter reproductive arrest period. LPII was also absent from young females that had received androgenic gland implants. LPII also appeared in the hemolymph of intersex individuals from which the androgenic gland had been removed. It is therefore suggested that LPII serves as a marker indicating the onset of secondary vitellogenesis in C. quad'iariicarintus females.

Soluble polysaccharide and biomass of red microalga Porphyridium sp. alter intestinal morphology and reduce serum cholesterol in rats.

The present study investigated the effects of the red microalga Porphyridium sp. on gastrointestinal physiology and lipid metabolism in male Sprague-Dawley rats. Diets containing dietary fibre from pelleted red microalgal cells (biomass) or their sulfated polysaccharide, pectin or cellulose (control) were fed to rats for a period of 30 d. All three fibre-supplemented diets increased the length of both the small intestine and colon, with a significantly greater effect in rats fed the algal polysaccharide. The polysaccharide also increased mucosa and muscularis cross-sectional area of the jejunum, and caused hypertrophy in the muscularis layer. The algal biomass significantly lowered gastrointestinal transit time by 44% in comparison with the control rats. Serum and mucosal cholecystokinin levels were lower in rats on the pectin and polysaccharide diets, while cholecystokinin levels in rats fed algal biomass were not different from those in the control animals. In comparison with the control diet, all the experimental diets significantly lowered serum cholesterol levels (22-29%). Feeding of non-fermentable algal polysaccharide or biomass significantly increased faecal weight and bile acid excretion compared with pectin-fed or control rats. The algal polysaccharide and biomass were thus shown to be potent hypocholesterolaemic agents active at low concentrations in the diet. Both metabolic and morphological changes were observed following consumption of algae, suggesting several possible mechanisms by which the alga affects lipid metabolism. The results presented in the present study encourage the use of red microalga as a functional food.

Prostaglandin E2 in previtellogenic ovaries of the prawn Macrobrachium rosenbergii: synthesis and effect on the level of cAMP.

Eicosanoids are thought to play a role in the regulation of invertebrate reproduction, as they do in vertebrate systems. This was investigated using the previtellogenic ovary of the freshwater prawn Macrobrachium rosenbergii as a biological model. Concentrations of prostaglandin E2 (PGE2), assessed by means of radioimmunoassay, in the previtellogenic ovary (oocyte diameter 20-40 microns) were 32.4 +/- 14.1 pg/mg ovary. Preincubation of the ovary with indomethacin (10 microM) inhibited PGE2 synthesis by 43%. In addition, if indomethacin was added to the culture medium, cAMP levels decreased by 48%. When previtellogenic ovaries were incubated in vitro with PGE2 (0.05 micrograms/ml medium and up), cAMP levels in the tissue homogenate sharply increased. The levels of cAMP rose most significantly (up to 10-fold) when 1-10 micrograms PGE2/ml medium was applied. These results suggest that PGE2, and possibly other prostaglandins, may play a role in the endocrine regulation of crustacean reproduction.

Enhanced 2-deoxy-D-glucose uptake and metabolism in splenocytes from diabetic and diabetes-prone BB rats. Further evidence to support prior in vivo activation.

Glucose metabolism in splenocytes from the BB rat was studied for the presence of abnormalities in [14C] 2-deoxy-D-glucose (2-dGlc) uptake, [U-14C]glucose conversion to 14CO2, and the production of lactate and pyruvate. Cells were studied freshly isolated ("resting"), and following culture both unstimulated (control) and stimulated with concanavalin A (ConA) or phorbol myristate acetate (PMA) + ionomycin. Both resting and control cells from diabetic (BBd) and diabetes-prone (BBdp) rats transported more (p less than 0.05) 2-dGlc than did cells from nondiabetes-prone (BBn) rats. Consistent with prior in vivo activation, sustained in vitro, lactate production was higher (p less than 0.05) under control conditions in BBd and BBdp than in BBn cells. Lactate production increased less with ConA and PMA + ionomycin in both BBd and BBdp than in BBn cells. PMA + ionomycin increased 2-dGlc uptake as much in BBd and BBdp cells as in BBn cells. Elevated rates of pyruvate production were observed in BBd cells under resting, control, and (especially) ConA conditions, suggesting an abnormality in pyruvate conversion to lactate. Few changes were observed in 14CO2 production. The presence of similar abnormalities in BBdp cells to those of the BBd cells suggests that the diabetic state is not causal, and the absence of an in vitro effect of 15 mmol/liter glucose in BBn cells further tends to exclude hyperglycemia as a cause of these alterations.

Neonatal low-dose gamma irradiation-induced impaired fertility in mature rats.

The reproductive capacity of mature rats at the age of 8 days was studied following neonatal exposure to 0.06 Gy dose of gamma-radiation. Decreased litter size and reduced body weight of the pups on weaning day, but not at parturition, were observed in female rats. The reduced litter size was not associated with impaired ovulation, impaired uterine implantation or mortality in utero, but resulted from increased death rate or at near parturition. Of the neonatally irradiated males 29% were found to be sterile and had degenerated or necrotic testes. The testicular damage and the reduced growth rate of the offspring of the irradiated females demonstrate the extreme sensitivity of the immature reproductive system to ionizing radiation, even at very low doses.

The reproductive system of male rats exposed to very low doses of ionizing radiation, 1: Hormonal profile of animals exposed after sexual maturity.

Adult male rats were treated with a single, whole body exposure to a dose of 0.001, 0.01 or 0.1 Gy (1 Gy = 100 rad). The animals were sacrificed 1, 7, 30, 100 or 180 days following exposure. Serum testosterone, serum and hypophyseal LH, FSH and prolactin as well as hypothalamic LHRH were measured. In the 0.1 Gy irradiated rats, serum LH and FSH were 100% higher and serum prolactin was 30% lower as compared to controls, 30 days after irradiation. The concentrations of these hormones remained significantly different from control values even after 180 days after exposure. On the other hand, the hypophyseal content of LH, FSH and prolactin decreased in the 0.1 Gy treated rats, 100 days after irradiation. In the group of the 0.01 Gy irradiated rats, serum FSH increased by 70% and hypophyseal FSH decreased. No changes were observed in serum testosterone and hypothalamic LHRH in any of the irradiated groups and no changes were observed in the group exposed to 0.001 Gy. It may be concluded that a long term damage to the reproductive system of the male rat was induced by a single, whole body exposure to 0.1 Gy of gamma radiation.

Immunological responses to chronic heat exposure and food restriction in rats.

Immunological variables were studied in rats chronically exposed to high environmental temperature (35 degrees C). Responses were compared with those of rats at 25 degrees C both fed ad libitum and pair fed to the decreased intake found in heat-exposed rats. Heat-exposed rats showed slower delayed-type hypersensitivity responses to keyhole limpet hemocyanin. They showed lower counts of peripheral blood total T cells (OX19+) as well as helper T cells (W3/25+) and smaller numbers of splenic T cells. The thymus was decreased in size. Increased levels of serum IgG antitetanus toxoid antibodies were found in heat-exposed rats. [3H]-thymidine incorporation into Concanavalin A (ConA)-stimulated splenic lymphocytes was decreased in pair-fed rats but not significantly altered in heat-exposed rats compared with controls. Heat exposure alters some aspects of both cellular and humoral immune function in a manner different from that induced by comparable food restriction without heat exposure.

The effect of dibromochloropropane (DBCP) on in vitro cyclic AMP levels and testosterone production in rat testes.

Adult male rats were injected s.c. once a week for 3 weeks with DBCP, 20 mg/kg BW. Animals were sacrified 20 weeks after last injection. Body and testes weights were recorded and testes were taken for standard histological preparation and for in vitro experiments. The in vitro experiments were carried out on testes slices (90-110 mg) incubated for 3 h with or without the addition of hCG to the incubation medium. Cyclic AMP content of the tissue as well as testosterone released into the incubation medium were determined. Testes weights of DBCP treated animals were 68% lower than that of controls. All semiferous tubules were damaged and shrunken, thus, their number per microscope field was 2.6 times that of controls. Cyclic AMP levels in testes slices were similar in both DBCP treated and controls. The addition of hCG stimulated cyclic AMP accumulation to a much higher level in the DBCP treated than in controls. When calculated per one pair of testes the content in unstimulated pair was more than twice that of DBCP treated. Stimulation of hCG increased both DBCP treated and controls to similar levels. Testosterone release into the medium by slices was higher in DBCP treated than in controls and so was also the increment due to hCG stimulation. Similar results were obtained when testosterone release was calculated per one pair of testes. It is suggested that since the major testicular compartment damaged by DBCP is the tubular one, the proportion of the interstitium per testicular unit weight is larger than in controls, thus, cyclic AMP content increment due to hCG stimulation is much higher.(ABSTRACT TRUNCATED AT 250 WORDS)

Long term effects of dibromochloropropane (DBCP) on male rats' reproductive system.

Adult male rats were injected s.c. once a week for 3 weeks with DBCP, 20 mg/kg B.W. Animals were sacrificed 5, 9, 13, 17, 25 and 50 weeks after last injection. Body weight was recorded once a week. Prior to sacrifice each male was presented with proestral females in order to determine the male's mating behaviour and fertility. Testes were removed, weighed and taken for standard histological examination. DBCP treatment caused a reduction of body weight which reverted back to control levels some 17 weeks post injection. Testes weights were reduced and remained low despite the recovery of body weight. Generally, all males showed normal mating behaviour but most of them were infertile. Testicular histology showed a correlation between decreasing testicular weight and increasing percentage of degenerated seminiferous tubules, which was on the other hand correlated with decreasing tubular diameter. Serum levels of FSH and LH were significantly increased in the infertile DBCP treated males while values for the fertile ones were similar to those of controls. There were no differences in serum testosterone levels between DBCP treated and control animals. It is concluded that in DBCP treated rats testicular degenerative damages are associated with increased circulating gonadotrophin levels and with normal testosterone levels. Although mating behaviour is unaffected fertility is depressed and does not recover for at least 50 weeks post injection. It is suggested that DBCP treatment affects mainly the activity of the Sertoli cells while the Leydig cells are affected to a much lesser degree.

Insulin binding and glucose transport in adipocytes in neonatal streptozocin-injected rat model of diabetes mellitus.

The neonatal streptozocin (STZ)-injected rat (NSIR) model of diabetes mellitus resembles human non-insulin-dependent diabetes mellitus (NIDDM) with respect to abnormalities in insulin secretory responses. The suggestion that insulin deficiency leads to insulin resistance, a prominent feature of human NIDDM, led us to examine insulin binding and glucose transport in the NSIR during the development of hyperglycemia. Male Wistar rats were injected at 2 days of age with STZ (90 mg/kg i.p.) or vehicle alone. Mild insulin deficiency, reflected by minimally decreased fed plasma insulin concentrations, was apparent at 4 wk (mean +/- SE, control vs. NSIR, 2.32 +/- 0.19 vs. 1.75 +/- 0.21 ng/ml) and at 8 wk. Pancreatic insulin content was dramatically reduced in NSIR to 12 and 5% of control values at 4 and 8 wk, respectively (P less than .001). Fed plasma glucose concentrations increased in the NSIR between 4 and 5 wk and were significantly elevated at 8 wk (251 +/- 25 vs. 527 +/- 52 mg/dl, P less than .001). 125l-labeled insulin binding showed a progressive increase as a function of adipocyte volume in control and NSIR. Epididymal fat pad weights and adipocyte volumes were significantly decreased in the NSIR. Thus, insulin binding did not differ when expressed per cell number but was increased in NSIR when corrected for cell size (percent specific binding X 10(2), 8.49 +/- 0.96 vs. 11.56 +/- 1.08/microliter cell vol; P less than .05, all ages combined).(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of in vivo administration of 3-isobutyl 1-methyl xanthine (IBMX) on cyclic AMP levels, and DNA synthesis in Ehrlich ascites tumor cells.

The effect of isobutyl 1-methyl xanthine (IBMX) upon cyclic AMP level and DNA synthesis was determined in vivo in Ehrlich ascites tumor (EAT) cells. Cyclic AMP was found to be elevated by IBMX while 3H-thymidine incorporation into the cells was inhibited when the phosphodiesterase inhibitor was injected in the mice bearing the tumor cells. The effect upon cyclic AMP level and H thymidine incorporation was more pronounced in cells obtained 4 days after the implantation compared to the effect found 8 days after cells implantation. Following 12 days of implantation, a weaker effect was found upon cyclic AMP level and no effect was found on 3H-thymidine incorporation.

Physical activity and blood pressure in normotensive young women.

Blood pressure and heart rate were measured in 22 physically active normotensive women, 20 +/- 1.7 years old (mean +/- SD) and in 25 physically non-active normotensive women 20.9 +/- 1.7 years old in supine, sitting and standing positions. Diastolic and mean blood pressures were found to be significantly lower in active women than non-active women. A significant negative correlation was found between maximal oxygen consumption and blood pressure, and it is suggested that aerobic physical activity could be used as non-pharmacological therapy in cases of moderate and borderline hypertension in young women.

The role of calcium in the induction of refractoriness to cyclic AMP stimulation by TSH.

An initial exposure of beef thyroid slices to 25 mU/mL thyroid-stimulating hormone (TSH) for two hours induces a diminished stimulation of cyclic adenosine monophosphate (AMP) production upon subsequent readdition of TSH but does not modify the effect of prostaglandin E1 (PGE1). Incubation of thyroid slices in calcium-free buffer with or without 2 mmol/L ethylene glycol bis (beta-aminoethyl ether)--N,N' = tetracetic acid (EGTA) prevented desensitization induced by TSH and PGE1, to the subsequent stimulation by TSH and PGE1, respectively, despite the presence of calcium in subsequent incubations. TSH-induced desensitization was not modified by increasing the calcium concentration up to 50 mmol/L in the initial incubation. However, the stimulatory effect of TSH upon cyclic AMP levels was decreased as the calcium concentration in the first incubation was increased. In the presence of at least 1 mmol/L calcium, an initial incubation of thyroid slices with 20 mumol/L ionophore A-23187 decreased the stimulation of cyclic AMP by 25 mU/mL TSH added to the slices for the first time during a subsequent incubation. Under these conditions, A-23187 had no effect on PGE1 stimulation of cyclic AMP. These results indicate that calcium may play a role in the TSH-induced, but not PGE1, desensitization of cyclic AMP formation.

Effect of acute and chronic exposure to heat on pharmacokinetic parameters of digoxin in rats.

Digoxin pharmacokinetic parameters in acute heat-exposed male rats (AHE) maintained for 3 hours at 35 degrees C, and chronic heat-exposed rats (CHE) maintained at 35 degrees C for 4 weeks, were compared to control rats (C) maintained at 22 degrees C. Digoxin concentration in the plasma was measured by radioimmunoassay which gave values similar to thin layer chromatography method. The half life time of digoxin during the elimination phase of the drug (t 1/2 beta) was similar in all three experimental groups (2.8; 2.7; 3.0 hrs) while the values obtained by extrapolation of digoxin plasma concentration to zero time (CpO) was 40% higher in AHE rats and 375% higher in CHE rats than in C rats. The apparent distribution volume of the drug (Vapr) was 40% lower in AHE rats and 80% lower in CHE rats as compared to C rats.

Renal clearance of urea, inulin, and p-aminohippurate in heat-acclimated rats.

Clearance values of urea, inulin, and p-aminohippurate (PAH) were measured in heat-acclimated (HA) rats exposed for 3 wk to 35 degrees C, in control rats (C) exposed to 23 degrees C, and in HA rats deacclimated (DA) for 3 wk. In HA rats, urea clearance was lower by 73%, inulin clearance by 61%, and PAH clearance by 56%, compared with C rats. The clearance values of these substances returned to the control values in DA rats. The data suggest that the low clearance values reflect both reduced renal blood flow and possible changes in the capacity of the glomeruli for filtration and the tubules for either reabsorption or secretion.

Glucoregulatory and metabolic responses to heat exposure in rats.

To determine the possible role of altered secretion and effects of insulin in fuel homeostasis during heat exposure, the hormonal and metabolic milieu of three groups of rats were studied. The first was placed at 35 degrees C for 12 days (HE), the second was pair-fed (PF) to the first but maintained at 23 degrees C, and the third was allowed to eat ad libitum at 23 degrees C (C). Plasma insulin, glucagon, glucose, and free fatty acids (FFA), and blood lactate, pyruvate, 3-hydroxybutyrate, and individual amino acids were determined. To further characterize glucoregulation, an intraperitoneal glucose tolerance test (1 mg/g body wt) and isotopic glucose turnover (primed infusion of [3-3H]glucose) were performed. In HE rats, weight was constant for the last third of the period, and metabolic state 4 h after food removal was characterized by euglycemia but hypoinsulinemia, elevated blood pyruvate and FFA, and normal 3-hydroxybutyrate compared with C. Lowered levels of branched-chain amino acids and arginine were found. Fourteen hours after food removal glucose turnover was decreased. However, glucose intolerance accompanied by hyperinsulinemia was also found. Many of these changes were also seen in PF, including constant weight, fasting euglycemia, hypoinsulinemia, elevated FFA, and lowered valine and isoleucine. In contrast, pyruvate concentrations were normal, that of 3-hydroxybutyrate was elevated, and the decrement in glucose turnover was smaller than in HE rats. The glucose tolerance was similar to that of HE but accompanied by hypoinsulinemia. The results in HE suggest decreased energy metabolism, insulin secretion altered in a complex manner, and altered insulin action.(ABSTRACT TRUNCATED AT 250 WORDS)

Role of calcium in acetylcholine-induced desensitization in dog thyroid slices.

Incubation of dog thyroid slices with 1 microM acetylcholine (ACH) for 3 h followed by a second 2-h incubation without it induces a diminution of stimulation of glucose oxidation by ACH during a third incubation of 45 min. Using a calcium-free medium during all incubations prevents the desensitization and reduces, but does not abolish, ACH stimulation of glucose oxidation. EGTA [ethylene glycol bis(beta-aminoethyl ether)-N,N'-tetraacetic acid] (2 mM) added to the calcium-free medium in all incubations prevents both refractoriness and stimulation of glucose oxidation induced by ACH. Calcium depletion during the first incubation only, achieved by using EGTA and a calcium-free medium, also prevents refractoriness but not the augmentation of glucose oxidation caused by ACH. Incubation of thyroid slices with 1 microM ionophore A23817 during the 3-h first incubation decreases the stimulation of glucose oxidation induced by its readdition or by 1 microM ACH added for the first time in the third incubation. Ionophore-induced desensitization is not related to a cholinergic muscarinic receptor effect. Initial incubation of dog thyroid slices with 1 microM ACH diminishes the subsequent stimulation of glucose oxidation by 0.5 microM ionophore. However, the ACH-induced desensitization to ionophore can be overcome by a 10-fold increase in the amount of ionophore in the third incubation. Ionophore (1 microM) in the first incubation also induces refractoriness to thyroid-stimulating hormone (TSH) (10 mU/ml)-stimulated glucose oxidation in the third incubation. In contrast, initial incubation of thyroid slices with TSH (25 mU/ml) does not affect the stimulation of glucose oxidation by 0.5 microM ionophore added during the third incubation. These results suggest that increased intracellular calcium plays a major role in, or even mediates, ACH-induced desensitization in the thyroid gland.

Exposure of dog thyroid slices to acetylcholine induces refractoriness to its subsequent stimulation of glucose oxidation.

An initial incubation of dog thyroid slices with 0.1 or 1 microM acetylcholine (ACH) for at least 2 h decreases its subsequent stimulation of [1-14C]glucose oxidation. Refractoriness persists for as long as 6 h in the absence of ACH. While new protein synthesis is essential for recovery, it is not necessary for its induction. Refractoriness is prevented when 25 microM tropicamide, an atropine-like drug, is present from the beginning of the initial incubation, but not when it is added after 2 h of incubation of slices with ACH, indicating that at this time ACH is no longer necessary for refractoriness. During refractoriness induced by ACH, stimulation of glucose oxidation by thyroid-stimulating hormone, prostaglandin E1, dibutyryl cyclic AMP, and cholera toxin, but not menadiol, is also significantly diminished. Incubation of thyroid slices with ACH does not modify its stimulation of iodide organification or 32Pi incorporation into phospholipids. These results suggest that the desensitization is not due to changes in the ACH receptor but rather to intracellular metabolic effects. This phenomenon may be important in the regulation of cholinergic effects on the thyroid.

The effect of prolonged exposure of pregnant rats to high ambient temperature on several constituents of the genital tract's fluids.

1. Various constituents of the genital tract's (GT) fluids were measured in heat-exposed rats (kept for at least 30 days at 35 +/- 1 degree C) and control rats (maintained at 22 +/- 2 degrees C). 2. There were no differences between the groups in the GT fluid volume, protein, free amino acids and glucose contents. 3. Arterial and venous blood glucose levels, pO2 and pH values were similar in both groups. 4. GT fluid protein hydrolysate from heat-exposed rats showed significantly reduced contents of glycine and alanine and elevated contents of valine and lysine as compared with controls. 5. The GT fluid's free amino acid components showed reduced glycine and elevated valine and isoleucine in the heat-exposed group as compared with controls. 6. Progesterone levels in GT fluid of heat-exposed rats was 60% higher than that of controls. 7. It is suggested that the higher progesterone concentration and the altered relative contents of several free amino acids with a possible change in the proteins of the GT fluid may affect the development of the embryo in heat-exposed rats.

Protein kinase activity in liver of heat-acclimated hamsters.

Protein kinase activity in incubated liver slices from 35 degrees C heat-acclimated (HA) hamsters was 70% higher than in similar slices from 23 degrees C control (C) hamsters. Adding glucagon to the incubation medium increased protein kinase activity by 65% in slices from C animals, but by only 30% in slices from HA animals. Binding of [3H]cAMP to proteins of a low-speed supernatant fraction of incubated and homogenized slices was 30% lower for HA than for C hamsters. For each acclimation group this binding was reduced 30% by incubation of the slices with glucagon. The activities of phosphorylase kinase, phosphorylase phosphatase, and phosphorylase alpha in slices incubated with or without glucagon did not differ between groups. Addition of glucagon increased phosphorylase kinase by 30% and phosphorylase alpha by 40% but caused no change in phosphorylase phosphatase activity. These results suggest that heat acclimation of the hamster increases the amount of a species of liver protein kinase that is different from the one that mediates the effect of glucagon on glycogenolysis.