Effect of different doses of water-soluble retinoic acid on the lipid and 3-beta-ol steroid dehydrogenase concentration in the interstitial tissue and supporting cells of the testes of mice

A study was made of the influence of various doses of water-soluble retinoic acid on the concentration of lipids and 3-beta-ol steroid dehydrogenase (SD) in the interstitial tissue (IT) and supporting cells (SC) of mouse testicles. Three groups of animals were injected with 1% retinoic acid i. p., with the doses injected being 0.1, 0.2 and 0.3 ml respectively. After the termination of experiments the animals were sacrificed and the cryostat sections 9 micron thick were stained for lipids and 3-beta-ol in the testicles. It was found that water-soluble retinoic acid is capable of exerting an essential influence on the steroid-producing system of the testicles. At the same time retinoic acid injected in the doses 0.1 and 0.2 ml causes an elevation of the content of lipids and 3-beta-ol SD in the IT and SC of the testes producing Leydig's cells that furnish the internal secretion of the testicle. Injection of 0.3 ml retinoic acid brings about a decrease in the concentration of 3-beta-ol SD in the IT and SC as compared with the dose 0.2 ml. However, in the IT the activity of the enzyme remains higher than in control, whereas in the SC it falls below the control values. The lipid content in the IT and SC changes differently, namely it rises in Leydig's cells and diminishes in Sertoli's cells as compared with the effect of 0.3 ml acid. It may be suggested that injection of large doses of retinoic acid entails alterations primarily in the synthesis of sex steroids, whereas the storage of lipoid substances that serve as substrate remains at a high enough level.     

A Nuclear Stain for Escherichia Coli And Related Organisms

The dye base of new fuchsin was precipitated by adding potassium hydroxide to the dye solution. The precipitate was filtered out and washed with water. It was then suspended in water, brought into solution and adjusted to a pH of about 5.0 with nitric acid. The staining solution was prepared by adding 0.3 ml. of a 14% aqueous solution of pyrogallol and 0.1 ml. of a 1% aqueous solution of boric acid to 3.0 ml. of the dye solution. Smears of cells were made in water on a slide and allowed to dry before covering with the staining solution which was also permitted to air dry. The smear was then washed in water and mordanted for 5-20 seconds in a 0.1% aqueous solution of mercuric nitrate. After rinsing in water, the smear was air dried. When dry, the slide was placed on a 50° C. warm plate for a few seconds before covering with a very thin film of a 5% aqueous solution of nigrosin which had a pH of about 5.0.     

The optimum time for single artificial insemination of blue fox vixens (Alopex lagopus) with frozen-thawed semen from silver foxes (Vulpes vulpes)

During the breeding season of 1991 a total of 608 blue fox vixens aged 1 to 6 years (2.3 +/- 0.1 years, mean +/- SEM) from 2 farms were artificially inseminated intrauterine once with frozen-thawed silver fox semen (1 ml dose containing a total of 150 million spermatozoa). The vixens were allocated to 3 different groups according to the time of insemination. Vixens in Group 1 (n = 203), Group 2 (n = 198), and Group 3 (n = 207) were inseminated on the first, second or third day after the peak value of vaginal electrical resistance, respectively. An overall conception rate of 75% (456 of 608) and 6.0 +/- 0.1 (mean +/- SEM) cubs per litter was obtained. Conception rates and mean litter sizes were significantly different between groups of vixens with respect to day of insemination (P = 0.02, Chisquare, Kruskall-Wallis Test). Vixens inseminated on the second day (Group 2) had the highest conception rate (81%) and the largest mean litter size (7.0 +/- 0.2 cubs) of the three groups, while those inseminated on the third day (Group 3) had the lowest conception rate and mean litter size (70%, 5.4 +/- 0.3 cubs).     

萘乙酸诱发小鼠睾丸生精细胞损伤作用的研究(英文)

目的:探讨萘乙酸(?α-naphthlcetic acid,NAA)对小鼠睾丸生精细胞的损伤作用。方法:将50只健康雄性昆明种小鼠随机分为5组,每组10只。正常对照组和阳性对照组小鼠喂饲普通颗粒饲料,高、中、低剂量组分别将5000、1000、200mg/kg的萘乙酸加入饲料中喂饲小鼠,阳性对照组于处死前3天每天腹腔注射环磷酰胺20mg/kg体重。4周后处死小鼠取睾丸组织,用MTT法测定各组睾丸生精细胞增殖活性,用免疫组织化学方法检测各组Bcl-2和半光天冬酶(Caspase-3)表达。结果:高剂量组和正常对照组的睾丸生精细胞增殖活性分别为0.464±0.022和0.866±0.024,差别有显著性意义(P<0.05)。高剂量NAA组Bcl-2和Caspase-3阳性表达率分别为11.4%和40.2%,正常对照组分别为30.6%和8.6%,经检验,差异均有显著性(P<0.05)。结论:萘乙酸能诱发睾丸生精细胞凋亡,对睾丸生精细胞增殖活性有抑制作用。     

Hyperoxia prevents carrageenan-induced enlargement of functional residual lung capacity in rats

Experimental pneumonia induced by intratracheal application of carrageenan or paraquat increases the functional residual lung capacity (FRC) in rats. The mechanism of this increase is not clear, but a decrease in PO(2) may be involved. To test this possibility, we attempted to eliminate the PO(2) decrease in carrageenan-treated rats by exposing them to hyperoxia. Animals of the first group were exposed to 7 days of hyperoxia (F(I)O(2) 0.78-0.84, group Car+O(2)) after intratracheal application of carrageenan (0.5 ml of 0.7 % carrageenan in saline), whereas animals of the second group were given the same dose of carrageenan but breathed air (group Car+A). The third group of rats was kept for seven days in hyperoxia (group O(2)) and the fourth group served as controls (C). The animals were then anesthetized and intubated and their ventilatory parameters and FRC were measured during air breathing. Carrageenan application induced a FRC increase (Car+A 2.0+/-0.2 ml, C 1.6+/-0.1 ml), which was not seen in carrageenan-treated rats exposed to hyperoxia (Car+O(2) 1.6+/-0.1 ml). Hyperoxia alone did not affect the value of FRC (O(2) 1.5+/-0.1 ml). These results support the hypothesis that a decrease in PO(2) plays an important role in the carrageenan-induced increase of FRC in rats.     

第三脑室注射组胺及其受体激动剂对五肽促胃液素诱导...

The present study shows the dual effects of intraventricularly injected histamine (0.25-2.0 micrograms/5 microliters) on pentagastrin-induced gastric acid secretion. Male Wistar rats weighing 200-300 g were anesthetized with intraperitoneal sodium pentobarbital. Gastric acid was continuously washed out with 37 degrees C saline solution by means of a perfusion pump. On the background of continuous intravenous infusion of pentagastrin [7.5 micrograms/(kg.h),] histamine (0.25 microgram/5 microliters) or 2-pyridylethylamine (PEA, 10 micrograms/5 microliters), a H1-receptor agonist, was injected into the third ventricle through a chronically implanted canula. The acid output decreased 10 min after injection and did not recover at 90 min. When the dose of histamine was increased to 1.0 micrograms or 2.0 micrograms, dual effects appeared. The acid output decreased respectively in 73% or 50% of the animals, while in the rest 27% and 50% of the animals, the acid output increased. H2-receptor agonist dimaprit (10 micrograms/5 microliters, i.c.v.) or impromidine (0.1 micrograms/5 microliters, i.c.v.) had no pronounced effect on pentagastrin-induced acid secretion. Pretreatment with diphenhydramine (16 micrograms/0.2 ml or 32 micrograms/0.2 ml, i.m.) abolished the inhibitory effect of histamine and PEA on acid secretion. These results suggest that histamine may be involved in the central regulation of gastric acid secretion, and the inhibitory effect may be mediated by H1-receptors in the brain. The mechanism underlying the production of the dual effects of histamine is unknown.     

Serial determination of lung volume in small animals by nitrogen washout

This report describes the design of an apparatus and the procedures used to serially measure the total lung capacity and the functional residual capacity of small animals utilizing the N2-washout technique. The calibration data indicate that the technique is accurate to within 1 ml and has a variance of less than 5%. The in vivo lung volume measurements of rats were validated by comparing them with values obtained with a water-displacement technique; the means were within 0.3 ml. Examples of the precision and changes in lung volume of animals during studies are included to demonstrate the reliability and usefulness of the technique.     

Ultrastructural changes in the spermatogenic epithelium and in the basic components of the blood-testis barrier in adult rats following repeated administration of estradiol dipropionate]

Electron microscopic studies of the testes were conducted in intact adult Wistar rats and in rats given repeated injections of a 0.1% solution of estradiol-dipropionate-0.3 mg week for 10 weeks. Administration of the hormone caused focal inhibition of spermatogenesis (6 weeks) accompanied by development of destructive changes in the spermatogenic cells. These pathological changes were reversible and disappeared two months after the cessation of the hormone injection, and the spermatogenesis was restored. Development of the pathological changes in the spermatogenic epithelium was accompanied by disturbances of the fine structure of the main components of the blood-test is barrier (Sertoli cells, tunica propria of the seminiferous tubules).     

Endothelin-1 in citric acid aerosol inhalation-induced airway constriction of guinea pigs

Endotheline-1 (ET-1) has been shown to enhance tachykinin-induced airway constriction. This study was designed to test whether ET-1 is involved in citric acid-induced bronchoconstriction. Forty-eight anesthetized-paralyzed guinea pigs were divided into six groups of 8 animals each: saline control; citric acid; ET-1; ET-1 + citric acid; BQ123 + ET-1 + citric acid; and BQ788 + ET-1 + citric acid. BQ123 and BQ788 are specific ETA and ETB receptor antagonists, respectively. Each animal in the saline control group received 50 breaths of 4 ml saline aerosol and in all citric acid-treated groups was given 50 breaths of 4 ml aerosol generated from 0.6 M citric acid. In all ET-1-treated groups, each animal was exposed to aerosol generated from 10(-8) M ET-1. The animal in the ET-1 + citric acid group was exposed to ET-1 5 min prior to the citric acid. For the last two groups, each animal was first exposed to aerosol generated from either 10(-5) M BQ123 or 10(-5) M BQ788. Five min later, the animal was exposed to ET-1; and then 5 min later was followed by citric acid. Dynamic respiratory compliance (Crs), forced expiratory volume in 0.1 sec (FEV(0.1)), and maximal expiratory flow at 30% total lung capacity (Vmax 30) were obtained before and 3-15 min after citric acid. Either citric acid or ET-1 inhalation caused significant decreases in Crs, FEV(0.1), and Vmax 30, indicating airway constriction. Citric acid-induced airway constriction, for most cases, was not significantly augmented by ET-1. However, either BQ123 or BQ 788 significantly attenuated the airway constriction induced by the combination of ET-1 and citric acid. Also, in an additional study, either BQ123 or BQ788 significantly attenuated citric acid-induced airway constriction. These data suggest that endogenous ET-1 plays an important role in citric acid aerosol-induced airway constriction in guinea pigs.     

Production and enhanced biological activity of a novel GHRH analog, hGHRH with an N-terminal Pro-Pro extension

Growth Hormone Releasing Hormone (GHRH) is one of the most important hormones in life. Because of its potential clinical importance, its short half-life, and its expensive chemical synthesis, an analog of hGHRH with a prolonged half-life and better activity has been studied for clinical application, especially for the treatment of muscle wasting, type II diabetes, or sleep disorders. The Pro-Pro-hGHRH(1-44) peptide has better activity. The fusion partner gene with 127 amino acid residues of the C-terminus from l-asparaginase was recombined with asp-pro-pro-hGHRH(1-44) gene synthesized by PCR method to form a fusion protein with the unique acid labile linker Asp-Pro. The recombinant protein was expressed to high levels in Escherichia coli BL21 (DE3). The Pro-Pro-hGHRH(1-44) peptide was purified to homogeneity by means of cell disruption, washing, ethanol precipitation, acid hydrolysis, and SP-Sephadex C-25, and Sephadex G-25 column chromatography. The fold of the purification was about 88 times and the yield was 1.1% of the total protein weight of the inclusion body. The peptide molecular mass of 5235.25 Da was determined by ESI mass spectroscopy. Its purity was determined by SDS-PAGE. In the study of the activity, we measured GH release of rat pituitary by using the antiserum kit against human GH. The peptide doses of 0.01, 0.1, 1.0, 7.72, and 20.9 microg/ml used, respectively, released the GH values of 0.1+/-0.1, 12.5+/-7.3, 16.6+/-5.8, 49.8+/-7.6, and 79.5+/-5.7 ng/ml whereas their blank controls, respectively, were 0.5+/-0.8, 4.1+/-2.6, 3.1+/-3.1, 4.7+/-1.8, and 1.2+/-0.3 ng/ml. The activity results of all dose groups except 0.01 microg/ml Pro-Pro-hGHRH(1-44) group and hGHRH(1-40) group showed that there were significant differences between GH released by the peptide and that by its blank control. With the increase of dosage, the differences were more significant. hGHRH(1-40) showed no measured GH release when the dose was up to 2 microg/ml. The activity results show that the Pro-Pro-hGHRH(1-44) peptide is a potential GH releasing analog.     

Direct effects of heparin on basal and stimulated aldosterone production in rat adrenal glomerulosa cells

Direct effects of heparin (0.1-10 IU/ml) on basal and stimulated aldosterone production have been studied using intact rat adrenal glomerulosa cells. Heparin at any dose did not affect basal aldosterone production when added to the incubation medium. Heparin at a 0.01 IU/ml dose had no effect on aldosterone production maximally stimulated by angiotensin II (AII, 4.8 X 10(-8) M), ACTH (4.3 X 10(-9) M) or potassium (8.0 mM). However, heparin at 0.1 and 0.3 IU/ml doses selectively blocked aldosterone production maximally stimulated by AII but not by ACTH or potassium, while the compound at 1 and 10 IU/ml doses inhibited aldosterone production maximally stimulated by these three stimuli. In addition, the inhibitory effect of 0.3 IU/ml heparin occurred as early as 30 min after incubation with heparin. These data suggest that heparin at 0.1 and 0.3 IU/ml doses acts directly on adrenal zona glomerulosa to selectively block the stimulatory action of AII, while the compound at 1 and 10 IU/ml doses inhibits all the stimulatory actions of AII, ACTH and potassium.     

Effect of caffeic acid phenethyl ester on treatment of experimentally induced methicillin-resistant Staphylococcus epidermidis endophthalmitis in a rabbit model

This study investigated the anti-inflammatory effects of caffeic acid phenethyl ester (CAPE), a natural bee-produced compound, and compared it with corticosteroids in the treatment of experimentally induced methicillin-resistant Staphylococcus epidermidis (MRSE) endophthalmitis in addition to intravitreal antibiotics. An experimental endophthalmitis model was produced in 24 New Zealand albino rabbits by unilateral intravitreal injection of 0.1 ml of 4.7 x 10(4) colony-forming units (CFU) methicillin-resistant S. epidermidis. The animals were then divided randomly into three treatment groups and a control group, group 1 (six rabbits), received only intravitreal vancomycin (1.0 mg/0.1 ml); group 2 (six rabbits), received both intravitreal vancomycin (1.0 mg/0.1 ml) and intravitreal dexamethasone (400 microg/0.1 ml) and group 3 (six rabbits), received both intravitreal vancomycin (1.0 mg/0.1 ml) and subtenon CAPE (10 mg/0.3 ml) after 24 h post-infection. No treatment was given to the control group. Treatment efficacy was assessed by clinical examination, vitreous culture and histopathology. There were no statististically significant differences between clinical scores of all groups in examinations at 24 and 48 h post-infection (p = 0.915 and p = 0.067 respectively), but in examinations at 72 h post-infection and after 7 days post-infection, although the clinical scores of treatment groups were not significantly different from each other, they were significantly lower than the control group (p < 0.05). The culture results of all groups were sterile. As a result, CAPE was found to be as effective as dexamethasone in reducing inflammation in the treatment of experimental MRSE endophthalmitis when used with antibiotics. More studies are needed to determine the optimal administration route and effective dosage of this compound.     

Sensitivity of aflatoxin b1 to ionizing radiation.

Aflatoxin B1 in a 5-micrograms/ml water solution was sensitive to ionizing radiation. Inactivation was assayed by the Ames microsome mutagenicity test and confirmed by thin-layer chromatography. Destruction of aflatoxin B1 had already begun at 2.5 kilograms (kGy; 1 kGy = 0.1 Mrad), but a dose exceeding 10 kGy was necessary for total destruction.     

Analyses of sister-chromatid exchange and cell-cycle kinetics in mouse T- and B-lymphocytes from peripheral blood cultures

A reliable mouse peripheral blood lymphocyte culture assay has been developed for sister-chromatid exchange analysis. Crucial aspects for optimal mitogenesis include: (1) the addition of 5 X 10(5) leucocytes/ml culture; (2) the use of animals with leucocyte counts from 5 to 7 X 10(6)/ml; and (3) the addition of 6% mouse plasma for the first 24 h of a total 54-h incubation. When 7 micrograms phytohemagglutinin/ml were used to stimulate T-lymphocytes, the mitotic index was 3.4 +/- 0.3%, 28 +/- 2.3% of the metaphases were in first-division, and the SCE frequency/metaphase was 7.3 +/- 0.2 (n = 14 mice). When B-lymphocytes were stimulated with 60 micrograms lipopolysaccharide/ml, the mitotic index was 4.5 +/- 0.3%, 64 +/- 3.3% of the metaphases were in first-division, and the SCE frequency/metaphase was 4.6 +/- 0.1 (n = 7 mice). This culture method consistently yields sufficient numbers of metaphases from both B- and T-lymphocytes for SCE and chromosome-breakage studies.     

Sensitivity of aflatoxin b1 to ionizing radiation

Aflatoxin B1 in a 5-micrograms/ml water solution was sensitive to ionizing radiation. Inactivation was assayed by the Ames microsome mutagenicity test and confirmed by thin-layer chromatography. Destruction of aflatoxin B1 had already begun at 2.5 kilograms (kGy; 1 kGy = 0.1 Mrad), but a dose exceeding 10 kGy was necessary for total destruction.     

Effect of estradiol on tissue glycogen metabolism in exercised oophorectomized rats

The effect of both physiological and pharmacological doses of estradiol on exercise performance and tissue glycogen utilization was determined in oophorectomized estradiol-replaced (ER) rats. Doses of beta-estradiol 3-benzoate (0.02, 0.04, 0.1, 0.2, 1, 2, 4, or 10 micrograms.0.1 ml of sunflower oil-1.100 g body wt-1) were injected 5 days/wk for 4 wk. Controls were sham injected (SI). After treatment, the animals were run to exhaustion on a motorized treadmill. ER animals receiving the 0.02-microgram dose ran significantly longer and completed more total work than the SI group. ER animals receiving doses of greater than or equal to 0.04 microgram ran longer and performed more work than the 0.02-microgram group. At exhaustion, myocardial glycogen content was significantly decreased in animals that were ER with less than or equal to 0.1 microgram, whereas those replaced with doses greater than 0.1 microgram utilized significantly less glycogen. With the 10-micrograms dose no significant decrease in heart glycogen content was observed at exhaustion. A submaximal 2-h run significantly reduced glycogen content in heart, red and white portions of the vastus lateralis, and the livers of SI animals. The latter effect was attenuated in skeletal muscle and liver, and there was no effect in the hearts of the ER animals receiving 2 micrograms. These data indicate that estradiol replacement in oophorectomized rats influenced myocardial glycogen utilization during exhaustive exercise and spared tissue glycogen during submaximal exercise. These glycogen sparing effects may have contributed to the significant improvements in exercise performance observed in this study.     

Rapid calorie metering inad lib rats

Adult wistar rats of either sex housed in individual cages and fedad lib were used to investigate the effects of calorie content and taste on ingestion. A 15 min single-choice solution (water, quinine, sucrose, saccharin) test as well as 1 h mixed diet (stock diet, sucrose-mixed, saccharin-mixed, quinine-mixed) tests were used. Calorically-rich sucrose either in solution or in mixed-diet was preferentially ingested. Tasteper se has not influenced calorie intake, as intake on sweet saccharin (3.4 ±0.4 cal) or on bitter quinine (3.3 ±0.6 cal) was similar to intake on stock diet (4.8 ±0.8 cal). Increased 5 min intake on sucrose solution (4.6 ±0.1 ml) over intake of other test solutions (saccharin 3.5 ±0.2 ml, quinine, 1.2 ±0.3 ml, water 2.1 ±0.1 ml) and calorie intake suppression on 1 h stock diet immediately following sucrose solution intake, indicate rapid calorie metering, probably based on fast-acting specific gustatory signals     

A scorpion venom peptide fraction induced prostaglandin biosynthesis in guinea pig kidneys: incorporation of 14C-linoleic acid

A peptide fraction isolated from the venom of the Egyptian scorpion Buthus occitanus was proved to have a bradykinin- potentiating activity. In vivo and in vitro modes of action of the isolated bradykinin-potentiating peptide (BPP) on kidneys of guinea pigs were investigated. Animals received five successive i.p. doses of the scorpion BPP (1 microg/g body weight) at one-week intervals. The control animals were i.p. injected with saline solution only. In vivo experiments showed a significant increase in renal tissue PGE(2) content and lipid peroxides of the treated guinea pigs compared to the control animals (p < 0.05). Nonsignificant changes were detected in the levels of tissue c-AMP and 5-nucleotidase activity (p > 0.05) of the treated animals, while the changes in c-GMP and c-AMP/c-GMP ratio were both significant (p < 0.05). In vitro experiments demonstrated enhanced capacity of guinea pig-renal tissue to convert (14)C-linoleic acid to its metabolites, 6-keto-PGF(1)alpha, PGF(2)alpha, PGE(2), TxB(2), PGD(2), and arachidonic acid, in response to the added PBP (1 microg/ml) and bradykinin (1 microg/ml). This enhanced response was abolished upon the addition of 1 microg/ml of BK-inhibitor (D-Arg- [Hyp(3), Thi(5,6), Phe(7)]). The capacity for labeled metabolites recovery in BPP treated renal tissue was 19.78%, while it was 13.00% in the basal control. The total increase that evoked by BPP was 62.78%. The results clearly indicate that the isolated BPP induced prostaglandin biosynthesis, which may trigger enhanced glomerular filtration in guinea pigs.     

Effects of synthetic atrial natriuretic factor (ANF) on renin-aldosterone axis in the conscious newborn calf

The effects of synthetic atrial natriuretic factor (ANF) on the renin-aldosterone axis were studied in fifteen 4-7 day-old male milk-fed calves divided into 3 groups of 5 animals each. Synthetic ANF intravenous (i.v.) administration (1.6 micrograms/kg body wt over 30 min) induced a transient significant fall in plasma renin activity (from 2.5 +/- 0.3 to 1.7 +/- 0.3 ng angiotensin l/ml/h; P less than 0.05) but failed to reduce basal plasma aldosterone levels in the first group of animals. Administration (i.v.) of angiotensin II (AII) (0.8 micrograms/kg body wt for 75 min) was accompanied by a progressive fall in plasma renin activity (from 2.2 +/- 0.3 to 0.8 +/- 0.1 ng angiotensin l/ml/h; P less than 0.01) and by an increase in plasma aldosterone levels (from 55 +/- 3 to 86 +/- 5 pg/ml; P less than 0.01) both in the second and the third groups; addition of ANF to AII infusion (AII: 0.5 mu/kg body wt for 45 min; AII: 0.3 micrograms/kg body wt and ANF 1.6 micrograms/kg body wt during 30 min) in the third group did not modify plasma renin activity or AII-stimulated plasma aldosterone levels when compared to the AII-treated group. These findings show that in the newborn calf ANF is able to reduce plasma renin activity but fails to affect basal and AII-stimulated plasma aldosterone levels, suggesting that the zona glomerulosa of the newborn adrenal cortex is insensitive to a diuretic, natriuretic and hypotensive dose of the atrial peptide.     

Involvement of angiotensin-mediated renal vasoconstriction in renal hypertension

Systematic arterial blood pressure and renal vascular resistance were found to be significantly greater in morphine, chloraloseurethane anesthetized renal hypertensive dogs than in similarly treated normotensive dogs. A lower dose infusion of the angiotensin antagonist 1-sar-8-ala-angiotensin II in the concentration of 20 mμg/ml into the renal artery decreased renal vascular resistance in the hypertensive, but not in the normotensive animals. The subsequent administration of a higher dose (approximately 50 mμg/ml) of 1-sar-8-ala-angiotensin II produced a decrease in renal vascular resistance in the normotensives, but a still greater effect in the hypertensives. Systemic blood pressure was significantly decreased with the higher dose in the hypertensive, but not in the normotensive group. The results indicate the participation of angiotensin-mediated renal vasoconstriction in the increased renal resistance in the hypertensive animals.