Gonadotropins and sex hormones modulate interrenal function in soft-shelled turtle

The aim of the current work was to investigate the role of gonadotropins and female sex hormones on interrenal activity in soft-shelled turtles, Lissemys punctata punctata. 1) FSH treatment (3 microg/100 g body wt daily for 10 days) caused interrenal hypertrophy with increased nuclear diameter, raises of acid phosphatase and alkaline phosphatase concentrations, and depletions of cholesterol (except the free fraction) and ascorbic acid levels from the interrenal gland. However, LH treatment (3 microg/100 g body wt daily for 10 days) failed to produce any perceptible change in the interrenal activity. The combined treatments of FSH and LH (3 microg each/100 gm body wt daily for 10 days) produced no further change beyond that of FSH alone. 2) Estrogen treatment with the low dose (25 microg/100 g body wt daily for 10 days) had no effect, but with higher doses (50 microg or 100 microg/100 gm body wt daily for 10 days) is caused interrenal stimulation by inducing the same manifestations to those of FSH. The degree of manifestations was higher with the higher dose (100 microg daily) than that of the moderate dose (50 microg daily). Progesterone treatment with the low dose (25 microg /100 g body wt daily for 10 days) had no significant effect, but with the moderate (50 microg daily) and higher (100 microg daily) doses suppressed interrenal activity by showing the reverse changes to those of estrogen. The degree of manifestations was higher with the higher dose than that of the moderate one. The combined treatments of estrogen and progesterone (100 microg each/100 g body wt daily for 10 days) caused interrenal stimulation but to a lesser extent than that of estrogen alone. The findings are briefly discussed.     

Adrenomedullary and glycemic responses to ACTH, corticosterone, aldosterone, epinephrine and norepinephrine administrations in the soft-shelled turtle

The aim of the current investigation was to ascertain the role of ACTH and adrenal hormones on adrenomedullary and glycemic functions in soft-shelled turtles, Lissemys punctata punctata. All the experiments were carried out on sexually immature animals. Findings revealed that: (1) ACTH administration (0.5 IU/1.0 IU/2.0 IU per 100 g body wt. daily for 10 days) in all doses stimulated adrenomedullary function by increasing medullary cell nuclear diameter with elevations of norepinephrine, epinephrine and blood sugar levels. Only moderate and higher doses (50 microg/100 microg per 100 g body wt. daily for 10 days) of dexamethasone suppressed adrenomedullary activity and blood sugar level by reversing the changes to those of ACTH; the responses were dose-dependent. But these changes were no longer observed after ACTH treatment in dexamethasone (DMS) recipients (DMS: 100 microg/ 100 g body wt daily for the first 10 days and ACTH: 0.5 IU / 100 g body wt daily for the next 10 days); (2) Only moderate and higher doses (50 microg/100 microg per 100 g body wt daily for 10 days) of corticosterone increased adrenomedullary activity and blood sugar level and the responses were also dose-dependent. But aldosterone treatment in all doses (same as for corticosterone) had no significant effect on the adrenal medulla or blood sugar level; (3) Only moderate and higher doses of norepinephrine or epinephrine (same as for corticosterone) caused adrenomedullary atrophy with depletions of norepinephrine and epinephrine levels but elevated the glycemic level. The findings are briefly discussed.     

Effect of pretreatment of Cassia fistula Linn. leaf extract against subacute CCl4 induced hepatotoxicity in rats

CCl4 alone treatment (0.lml of liquid paraffin/100g body weight, ip) for 7 days followed by 0.l ml of CCl4 (in liquid parafiin/100g body weight, ip) from day 8 till day 14, caused a 16 fold increase in lipid peroxidation and a 50% reduction in catalase and glutathione reductase in liver tissue of rats accompanied by an increase in the activities of transaminases. alkaline phosphatase, lactate dehydrogenase and gamma - glutamyl transpeptidase in serum as compared to liquid paraffin treated control. Pretreatment of ethanolic leaf extract of C. fistula (500mg/kg body weight/day for 7 days) followed by CCl4 treatment (0.1 ml/100g body weight from day 8 till day 14) completely reversed back lipid peroxidation and the activities of catalase and glutathione reductase in the liver tissue towards normalcy. This treatment also reversed the elevated levels of the enzymes in the serum. Ethanolic leaf extract alone treatment did not produce any change in all the parameters studied. The results suggest antioxidant and hepatoprotective properties of C. fistula during its pretreatment against CCl4 induced hepatotoxicity.     

Histochemical studies on the effect of thyroid hormone on alkaline and acid phosphatase activities in liver of fish and amphibia.

The Lata fishes (Ophicephalus punctatus) showed increased alkaline and acid phosphatase activities in liver after immersion for 15-30 days in thyroxine-containing medium (0.025 mug/ml). A single injection of thyroxine (1-2 mug/g of body weight) caused increased acid phosphatase activity in liver of Lata fish in comparison to the controls on the 5th day after experiment but the alkaline phosphatase activity remained unchanged. Both alkaline and acid phosphatases showed increased activities in liver of Lata fishes treated with a single injection of 4 mug of thyroxine per g of body weight on the 5th day. Immersion of Lata fishes in thiourea solution (1 mg/ml) for 15 days did not show any alteration in alkaline or acid phosphatase activities but these enzyme activities decreased after 30 days' immersion in thiourea solution in comparison to the controls. A seasonal variation of alkaline and acid phosphatase activities was observed in liver of Lata fishes. More alkaline phosphatase activity was found in liver of summer fishes than in winter fishes. The winter fishes showed more acid phosphatase activity than the summer fishes. Three consecutive injections of thyroxine (0.1 mug/g of body weight) to toads (Bufo melanostictus) caused increased alkaline and acid phosphatase activities in liver on the 5th day of the experiment, in comparison to the controls.     

Adrenal glucocorticoid function and cytoplasmic dehydrogenase activity in the presnece of mechanical and toxic liver damage]

Against the background of low steroid metabolism in the liver there was noted some decrease in the rate of corticosterone synthesis by the adrenal gland sections, and also, a decrease in the dehydrogenase activity in the cytoplasm of adrenal cells in male rats 48 hours after partial hepatectomy, as compared to the sham-operated animals. These changes resulted from suppression of the central mechanisms of stress realization due to the lowered steroid metabolism. Intraperitoneal administration of carbon tetrachloride (0.1 ml per 100 g of body weight) at the same periods led to a significant intensification of the steroidogenesis in the adrenal tissue and to the activation of NAD-dependent dehydrogenases in the cytoplasm. The role of toxic injury of the glands in the changes of the functional state of the adrenal cortex cells is discussed.     

Neurohypophysial-adrenal gland responses to water-deprivation in the rose-ringed parakeet.

The aim of the present investigation is to study the relative influence of neuohypophysis, adrenal cortex and adrenal medulla under dehydration stress in the parakeet. Birds subjected to dehydration for 7 days lost body weight. Neurosecretory material (NSM) was partially depleted from the neurohypophysis. Adrenal gland weight was increased followed by a hypertrophy of the cortical tissue. A fall in adrenal cholesterol and ascorbic acid level was marked. Adrenaline and noradrenaline contents of the adrenal medulla were suppressed as was evident from cytochemical and biochemical findings. It is suggested that neurohypophysis, adrenal cortex and adrenal medulla are involved in maintaining water homeostasis in this avian species.     

Acid and alkaline phosphatase activities in lithium treated testis, prostate and seminal vesicles of adult albino rats: evidence of duration and dose dependent response

Changes in the activities of acid and alkaline phosphatase were observed in the testis, prostate and seminal vesicle after the injection of lithium chloride at the doses of 100, 200 and 400 micrograms/100 g body weight/day for 7, 14 and 21 days. The studies indicate that 200 and 400 micrograms/100 g body weight for 14 days and 21 days showed a significant inhibition in the activity of acid phosphatase in all the above reproductive organs. There is a significant stimulation of alkaline phosphatase activity at the doses of 200 and 400 micrograms of lithium after 21 days of treatment in testis, prostate and seminal vesicle along with significant decrease in accessory sex organs weight in comparison to control animal. Therefore, it is evident that the effect of lithium on male reproductive organs mainly depends on the amount of the drug being injected and the duration of treatment to it.     

Water and sodium balance in the estuarine diamondback terrapin (Malaclemys)

Summary Total body water decreased significantly in terrapins exposed to sea water (SW). Although the intracellular fluid decreased somewhat upon SW exposure, the decline in extracellular fluid was almost twice as great. Under conditions of voluntary drinking after salt loading, terrapins substantially increased the volume of the extracellular fluid while maintaining the intracellular fluid near the freshwater (FW) control levels. FW terrapins were consistently heavier than animals of the same plastron length exposed to SW. Thus expression of body fluid volumes as ml/cm plastron length rather than as % body weight is necessary to correct for the loss of total body water with progressive dehydration. Fasted terrapins in SW lost weight at 0.32% weight/day, whereas the rate in FW was 0.21%/day. Water influx and efflux in SW were 0.17 and 0.16 ml/100 g·h respectively. When the efflux was increased by the calculated value for unmeasured respiratory loss, it exceeded the influex by 0.01 ml/100 g·h. Consequently the net water loss determined with radiotracers (equivalent to 0.24% weight/day) was similar to the difference between the weight losses in SW and FW (0.11%/day). Partitioning studies indicated that the majority of water exchange between the terrapin and SW occurs through the integument. Terrapins in SW underwent a concentration of the body fluids, most of which can be attributed to water loss, not electrolyte gain. The rates of Na influx and efflux were quite low (usually ranging from 6–10 moles/100 g·h). In two terrapins the injection of NaCl loads resulted in eight- to 19-fold increases in Na efflux. The uptake of Na from SW occurred orally. The skin was virtually impermeable to Na. The salt gland and possibly the cloaca were the major routes of Na efflux. The injection of NaCl loads resulted in an increase in cephalic Na excretion from a mean of 3.2 moles/100 g·h to 32.5 moles/100 g·h. Terrapins in SW exhibited a significant increase in bladder urine [K] over the FW controls. There was a direct relationship between plasma [Na], urine [K], and lachrymal salt gland Na–K ATPase content. In comparing SW terrapins with FW painted turtles (Chrysemys) exposed to SW radiotracer studies demonstrated a similarity in Na influx, but there was at least a four-fold increase in water exchange in the painted turtle. It seems likely that the skins of many aquatic reptiles (marine, estuarine and FW) are impermeable to Na but differ markedly in water permeability.     

Enzyme Histochemistry on Paraffin Embedded Tissue Sections

To obtain diagnostic enzyme reactions in paraffin embedded tissue sections, we compared four fixatives (buffered formol sucrose, Baker's formol calcium, periodate lysin paraformaldehyde, and buffered formalin acetone) and subsequent acetone dehydration with or without graded concentrations of Triton X-100. Four spleens and 14 lymph nodes were tested for peroxidase, naphthol ASD chloroacetate esterase, acid phosphatase, alpha naphthyl acetate esterase, and alpha naphthyl butyrate. Best results were obtained by a processing method using buffered formalin acetone, Holt's gum sucrose, dehydration in acetone with 0.03% Triton X-100, and paraffin for embedding.     

Effects of acute fresh water exposure on water flux rates and osmotic responses in Kemp's ridley sea turtles (Lepidochelys kempi)

Water flux rates and osmotic responses of Kemp's Ridley sea turtles (Lepidochelys kempi) acutely exposed to fresh water were quantified. Salt-water adapted turtles were exposed to fresh water for 4 d before being returned to salt water. During the initial salt water phase, absolute and relative water flux rates were 1.2+/-0.1 l d(-1) and 123.0+/-6.8 ml kg(-1) d(-1), respectively. When turtles were exposed to fresh water, rates increased by approximately 30%. Upon return to salt water, rates decreased to original levels. Plasma osmolality, Na(+), K(+), and Cl(-) decreased during exposure to fresh water, and subsequently increased during the return to salt water. The Na(+):K(+) ratio was elevated during the fresh water phase and subsequently decreased upon return to salt water. Aldosterone and corticosterone were not altered during exposure to fresh water. Elevated water flux rates during fresh water exposure reflected an increase in water consumption, resulting in a decrease in ionic and osmotic concentrations. The lack of a change in adrenocorticoids to acute fresh water exposure suggests that adrenal responsiveness to an hypo-osmotic environment may be delayed in marine turtles when compared to marine mammals.     

Hormone mediated changes in monoamine stores and regulation of enzymes of biosynthesis and metabolism in the rat adrenal gland. Influence of progesterone, estradiol, ACTH and testosterone administration

The influence of daily injections, for 7 days, of 17-hydroxyprogesterone caproate (8 mg/100 g body weight pro die), of estradiol-17 beta 17-undecylate (1.2 mg/100 g body weight pro die) of testosterone enanthate (2 mg/100 g body weight pro die) and of ACTH (4 IU/100 g body weight pro die) to male Wistar rats, weighing 200 g, were investigated. Monamine storage and regulation of enzymes phenylethanolamine N-methyl transferase (PNMT), monoamine oxidase (MAO) and catechol-O-methyl transferase (COMT) were studied in the adrenal glands. Results from testosterone treated rats were especially significant : the hormone administration decreased the stores of dopamine and norepinephrine (epinephrine precursors), stimulated PNMT and decreased metabolism of epinephrine by MAO and COMT. However, the adrenal content of epinephrine remained in the normal range. These findings suggest that uptake of epinephrine is accelerated in peripheral tissues by testosterone.     

Manifestations of social stress in grouped Japanese quail

Physiological manifestations of social stress in stable and unstable (resident and visitor) pairs of adult male Japanese quail (Coturnix coturnix japonica) were compared after 7, 14, 21 and 28 days of regrouping. Unstable pairs had reduced body and relative testes (mg/100 g) weights. Plasma cholesterol was increased significantly in unstable pairs, and adrenal cholesterol was reduced significantly by daily regrouping of unstable pairs. Twelve consecutive days of ACTH treatment (2, 4 or 8 IU/100 g body wt given intramuscularly) to birds in stable pairs induced a decrease in body and relative testes weights. The results were similar to those found in birds subjected to daily regrouping.     

Oral administration of phytocomponent p-hydroxycinnamic acid prevents bone loss in ovariectomized rats

The preventive effect of phytocomponent p-hydroxycinnamic acid (HCA) on ovariectomy (OVX)-induced bone loss was investigated. HCA (250 or 500 μg/100 g body weight) was orally administered once daily for 30 days to OVX rats. The analysis using a peripheral quantitative computed tomography (pQCT) showed that OVX caused bone loss in the femoral-metaphyseal tissues. This change was significantly restored after the administration of HCA (250 or 500 μg/100 g body weight) to OVX rats. Mineral content, mineral density, and polar strength strain index in the femoral-metaphyseal tissues were significantly decreased in OVX rats. These decreases were significantly restored after the administration of HCA (500 μg/100 g) to OVX rats. Moreover, OVX caused a significant decrease in calcium content or alkaline phosphatase activity in the femoral-diaphyseal and -metaphyseal tissues. These decreases were significantly restored after the administration of HCA (250 or 500 μg/100 g) to OVX rats. Deoxyribonucleic acid (DNA) content in the diaphyseal or metaphyseal tissues was significantly increased in OVX rats. These increases were significantly restored after oral administration of HCA (500 μg/100 g). This study demonstrates that HCA has preventive effects on OVX-induced bone loss of rats in vivo.     

Dependence of organism stability on chronic stress from thyroid status

In experiences on 108 male rats, the effect of acute (immobilization during 3 hrs) and chronic (immobilization for 3 hrs during 5 days) stresses on the organism general stability was studied as evaluated with changes of body weight, adrenal glands, spleen, thymus relative mass, gastric mucosa state, animals physical endurance. Chronic stress evoked more obvious decreasing of spleen and thymus relative mass than the acute one, as well as lesion of gastric mucosa accompanied with decrease of the rat resistance to physical loading. Thyroid function suppression by merkazolil (1.2 mg/100 g body weight during 14 days) promotes further the reduction of the organism stability in acute and, especially, in chronic stress, while physiological doses of thyroid hormones (5.0-8.0 mcg of thyroxin on kg of body weight during 28 days), on the contrary, increased it in both stress conditions. Existence of the organism stability dependence on thyroid status both in acute and chronic stress proves iodothyronine's important role in the organism antistress-system.     

Catecholamine hormones and the control of avian salt glands

Hypertonic saline loading (0.5 M NaCl, 15 ml.kg-1 i.v.) increased cardiac frequency and elicited nasal salt gland secretion in control ducks. Partial depletion of catecholamines by prior treatment with reserpine decreased body weight, lowered arterial pressure and abolished the tachycardiac responses to saline loading. Reserpine also increased plasma concentrations of Na, K and total osmolytes, yet altered neither the composition nor the flow rate of nasal fluid secretion. The preservation of the normal secretory responses to hypertonic stress in hypotensive, reserpine-treated ducks indicates that the nasal salt glands can function independently of changes in circulating catecholamine hormones.     

The hypothalamic and neurohypophysial vasopressin content as influenced by diphenylhydantoin in dehydrated rats.

Rats dehydrated up to 8 days were treated with diphenylhydantoin given intraperitoneally in daily doses of 10 mg/100 g of the initial body weight. The single dose of diphenylhydantoin diminished the vasopressin content in the hypothalamus and neurohypophysis of normally hydrated rats. Under conditions of severe dehydration (8 days), DPH treatment resulted in a more marked decrease of vasopressin in the hypothalamo-neurohypophysial system.     

Elevation of the panting threshold of the desert iguana,Dipsosaurus dorsalis,during dehydration: Potential roles of changes in plasma osmolality and body fluid volume

Summary Dehydration of the desert iguana,Dipsosaurus dorsalis, resulted in a progressive elevation in the magnitude of the skin temperature necessary to elicit thermal panting (i.e., the panting threshold). Panting threshold increased from 43.4±0.8 °C at 100% initial body weight (IBW) to 45.4±1.2 °C at 90% IBW to 45.7±0.9 °C at 80% IBW. Plasma osmolality showed no significant change with dehydration to 80% IBW. Changes in plasma osmolality, whether induced by NaCl or non-ionic sucrose loading, had a significant impact on panting threshold. Increasing plasma osmolality resulted in an elevation of panting threshold while decreasing plasma osmolality resulted in lower panting thresholds. Decreasing body fluid volume by exsanguination of 1 ml whole blood/100 g body weight resulted in a mean increase in panting threshold by 0.7±0.2 °C. Volume loading with 160 mM NaCl (approximately isosmotic) had no significant effect on panting threshold. These data suggest that plasma osmolality and decreases in body fluid volume may be potent modulators of panting threshold during periods of water deprivation. However, at least in desert iguanas, increases in plasma osmolality would not appear to be an important factor in the elevation of panting threshold during dehydration to 80% IBW.     

Kidney concentrating ability of a subterranean xeric rodent,the naked mole-rat (Heterocephalus glaber)

The naked mole-rat (Heterocephalus glaber) is a strictly subterranean mammal inhabiting the arid zones of north-east Africa. These animals have no access to free water and water balance thus might be facilitated by regulating renal water loss. The urinary concentrating ability of the naked mole-rat was determined using five dietary manipulations in which both water and salt content were altered. Control animals (n=7) received a high quality protein cereal mixed to a thin paste with water (1 g cereal: 85 g water). Water stress was induced by reducing the water content of the diet by either 50% (n=7) or 65% (n=7). Salt loading was facilitated by replacing the water with the same volume of either 0.9% salt (n=7) or 3.0% salt (n=4) solutions. Changes in body mass, food consumption and urine volume were measured daily. The effect of diet on osmolality and electrolyte concentrations of urine and plasma were determined on termination of the diet trials. Although energy intake was not reduced, naked mole-rats lost body weight with both water stress treatments. Urine volume voided per day decreased significantly with both water stress treatments (P<0.05), such that the most extreme water stress led to an 80% reduction in urine volume. Mildly salt-loaded animals gained weight, yet underwent a sodium diuresis, as indicated by a 1.3-fold increase in the daily volume of urine voided (P<0.05). Maximum urine concentration (1521±250 mmol·kg^-1) was achieved with mild water stress and was 4.6±0.9 times that of plasma. Neither further water stress nor salt loading further increased urine osmolality (P>0.05). The naked mole-rat exhibits a moderate kidney concentrating ability and cannot maintain plasma osmolality or body mass with either extreme water stress or salt loading. Although this species succesfully inhabits arid zones, survival in these areas is not facilitated by renal water conservation, but rather by their underground existence in a microhabitat where humidities are high and radiant heat loads low. In this milieu a moderate kidney concentrating ability is adequate.Abbreviations Bm body mass - ESL extreme salt load - EWS extreme water stress - MSL mild salt load - MWS mild water stress     

Randomised double blind trial of single dose doxycycline for treating cholera in adults.

OBJECTIVE--To compare the efficacy of a single dose of doxycycline (200 or 300 mg) with the standard multiple doses of tetracycline in patients with cholera. DESIGN--Randomised double blind controlled trial. Patients were given a single 200 mg dose of doxycycline, a single 300 mg dose of doxycycline, or multiple doses of tetracycline (500 mg, six hourly intervals). SETTING--Hospital in Bangladesh treating diarrhoea. PATIENTS--261 Patients aged over 15 admitted to the hospital with severe dehydration due to acute watery diarrhoea associated with Vibrio cholerae. All vibrios isolated from the stools and rectal swabs of patients, including those patients with prolonged excretion of vibrios, were sensitive to tetracycline. The stools of all patients at admission were negative for shigella and salmonella. INTERVENTIONS--All patients received rapid intravenous acetate solution for the first four hours after admission to hospital. They were then entered in the study and randomised. Oral rehydration was started immediately after the intravenous treatment. If signs of severe dehydration reappeared during oral treatment patients were given rapid intravenous acetate solution until dehydration was fully corrected. MAIN OUTCOME MEASURES--Stool output in first 24 hours and till diarrhoea stopped, total intake of oral rehydration fluid, duration of diarrhoea, and excretion of vibrio after receiving antibiotic treatment. RESULTS--The median stool outputs during the first 24 hours (275 ml/kg body weight) and till diarrhoea stopped (296 ml/kg body weight) were significantly higher in patients receiving 200 mg doxycycline as a single dose than in patients receiving either standard tetracycline (242 ml/kg body weight and 254 ml/kg body weight) or 300 mg doxycycline (226 ml/kg body weight and 255 ml/kg body weight). Similarly, median consumption of oral rehydration solution (18.45 l) was significantly higher in patients receiving 200 mg doxycycline than in patients receiving either 300 mg doxycycline (16.10 l) or standard tetracycline (14.80 l). Almost equal numbers of patients in each group required unscheduled intravenous acetate solution to correct dehydration during antibiotic treatment. Patients treated with doxycycline (low or high dose), however, had more prolonged excretion of bacteria. CONCLUSIONS--A single 300 mg dose of doxycycline is as effective as the standard multiple dose tetracycline treatment for cholera in terms of stool output, duration of diarrhoea, vomiting, and requirement for oral rehydration solution.     

Use of Gallocyanin as a Myelin Stain for Brain and Spinal Cord

Tissue fixed in 10% formalin, formol saline, CaCO₃ or phosphate buffer neutralized formalin, Baker's formol calcium, Cajal's formol ammonium bromide, formalin-95% ethanol 1:9, formalin-methanol 1:9, Lillie's methanol-chloroform or Salthouse's formol cetyltrimethylammonium bromide was dehydrated and embedded in paraffin. Sections were attached to slides with either albumen or gelatine adhesive and processed throughout at room temperature of 22-25 C. Mordanting 30-60 min in 1% iron alum was followed by a 10 min wash in 4 changes of distilled water. Myelin was stained in a gallocyanin self-differentiating solution for 1-2.5 hr; thick sections requiring the longer time. The staining solution (pH approximately 7.4) consisted of Na₂CO₃, 90 mg; distilled water, 100 ml; gallocyanin, 250 mg; and ethanol, 5 ml. The ethanol was added to this mixture last, and after the other ingredients had been boiled and then cooled to room temperature. After a staining and thorough washing, Nissl granules were stained for 5-10 min in a solution consisting of: 0.1 M acetic acid, 60 ml; 0.1 M sodium acetate, 40 ml; methyl green, 500 mg. Washing, dehydration, clearing and mounting completed the process. Myelin sheaths were stained dark violet; neuronal nuclei, light green with dark granules of chromatin; nucleoli of motor cells and erythrocytes, dark violet; cytoplasm, green with dark green Nissl granules. The simple and reliable method can be adapted easily for use with automatic tissue processors.