The functional morphology of the uterine capillary bed after synestrol administration

The capillary vessels of rats uterus was examined which were receiving for 7 days 0.2% solution of synoestrol i/m (2.0 mg per 1 kg of body weight). The animals were examined 1, 3, 5, 10, 20, 30, 60 days after the last injection of the solution. As the marker for the capillary vessels the histochemical method for finding the Mg2(+)-adenosintriphosphatase was used. The investigations have found prominent qualitative and quantitative changes (activity of enzyme, length and diameter of capillaries) not less than for two months after the last injection of the solution. Particularly prominent changes were found in endometrium between 10-20 days of the restorative period.     

Effect of ultrasound of high intensity on the intraorganic vessels of the liver]

Under study was the influence of a single 3-minute-long exposure to ultrasound of high intensity (2 wt/cm2) upon intraorganic vessels of the liver in albino rats. The circulatory bed was examined 10 min., 5, 15 and 30 days after treatment with ultrasound. Methods ox injection, rentgenography, claering and histology revealed an unfavourable effect of ultrasound causing local construction of arteries and veins in early terms, uneven filling of the capillary bed with the injected mass, dilatation and deformity of capillaries. The above changes grew more pronounced during 15 days, by the end of the month they diminished but not disappeared.     

Neuronal and Glial Expression of the Multidrug Resistance Gene Product in an Experimental Epilepsy Model

1. Failure of anticonvulsive drugs to prevent seizures is a common complication of epilepsy treatment known as drug-refractory epilepsy but their causes are not well understood. It is hypothesized that the multidrug resistance P-glycoprotein (Pgp-170), the product of the MDR-1 gene that is normally expressed in several excretory tissues including the blood brain barrier, may be participating in the refractory epilepsy. 2. Using two monoclonal antibodies against Pgp-170, we investigated the expression and cellular distribution of this protein in the rat brain during experimentally induced epilepsy. Repeated seizures were induced in male Wistar rats by daily administration of 3-mercaptopropionic acid (MP) 45 mg/kg i.p. for either 4 days (MP-4) or 7 days (MP-7). Control rats received an equivalent volume of vehicle. One day after the last injection, rats were sacrificed and brains were processed for immunohistochemistry for Pgp-170. As it was previously described, Pgp-170 immunostaining was observed in some brain capillary endothelial cells of animals from control group. 3. Increased Pgp-170 immunoreactivity was detected in MP-treated animals. Besides the Pgp-170 expressed in blood vessels, neuronal, and glial immunostaining was detected in hippocampus, striatum, and cerebral cortex of MP-treated rats. Pgp-170 immunolabeled neurons and glial cells were observed in a nonhomogeneous distribution. MP-4 animals presented a very prominent Pgp-170 immunostaining in the capillary endothelium, surrounding astrocytes and some neighboring neurons while MP-7 group showed increased neuronal labeling. 4. Our results demonstrate a selective increase in Pgp-170 immunoreactivity in the brain capillary endothelial cells, astrocytes, and neurons during repetitive MP-induced seizures. 5. The role for this Pgp-170 overexpression in endothelium and astrocytes as a clearance mechanism in the refractory epilepsy, and the consequences of neuronal Pgp-170 expression remain to be disclosed.     

Long-term enhancement of morphine hypalgesia as a result of periodic blockade of opiate receptors using naloxone

A significant enhancement of the analgetic effect of morphine (6 mg/kg, subcutaneously; tail withdrawal reflex at 60 degrees C) was observed in rats 3-4 hours after single naloxone (1 mg/kg) administration. Periodical naloxone injection (0.5 mg/kg, subcutaneously, 3 times per day at 3.5-hour intervals for 3 days) led to a prominent and long-term (testing on the 20th and 105th hour after the last naloxone administration) enhancement of morphine analgesia (2.6 mg/kg subcutaneously) and insignificant inhibition of stress analgesia during two-hour immobilization of animals. These modifications of morphine and stress analgetic effects are considered a result of adaptive changes of opiate receptors after their blockade.     

血红蛋白总量在运动员生物护照兴奋剂检测中的意义

目的：观察注射微量重组人促红细胞生成素（rHuEPO）后,受试者在不同时间段血液某些参数指标的变化,为兴奋剂rHuEPO检测运动员生物护照（ABP）提供实验依据。方法：采用单盲方式对14名健康青年男性受试者进行两个阶段随机安慰剂（n=14）和微量rHuEPO（n=14）皮下注射,每阶段7周,每周2次。EPO注射期间给予每天口服铁剂105 mg;在注射前第7日、第一次注射后第3、10、17、24、31、38和45日、以及第7周最后一次注射后第7、14、21日分别采取静脉血测试血液基础参数（红细胞、血红蛋白浓度[Hb]、红细胞压积、网织红细胞）,改良一氧化碳（CO）吸入法测试血红蛋白总量（tHb）以及全血量（BV）、血浆量（PV）的变化情况;分析这些指标在ABP中的使用,观察总量指标和浓度指标在检测中的有效性和时效性。结果：与安慰剂组相比,14名受试者在微量rHuEPO注射后第2周,即开始出现红细胞数量和[Hb]增加,在注射期间第5~6周达到高峰（分别增加9%和10%左右,P<0.01）,一直持续到最后一次注射后3周左右;tHb在注射后1周即开始增加,5周达到最高峰（增加10%左右,P<0.01）并一直持续到最后一次注射后1周才开始逐级恢复到注射前水平;红细胞总量在实验第5周上升至峰值,幅度为10.89%（P<0.01）,总血量（BV）变化不明显,血浆量（PV）出现下降,血液浓缩。结论：7周微量rHuEPO注射可以显著增加血液总量指标和浓度指标,其中血红蛋白总量更加敏感,可以用ABP血红蛋白总量等监测微量EPO注射;在最后一次注射结束后,血液总量指标如血红蛋白总量恢复正常,而浓度指标仍保持较高水平,可能是血液浓缩的结果。     

Vasculogenesis of the bursa cloacalis (bursa of Fabricius) of the chick embryo

Vasculogenesis of the bursa cloacalis (bursa of Fabricius) was examined in 10- to 21-day chick embryos and in chicks during the first 5 days post-hatching. The entire circulatory system was injected with India ink, and the bursae were then removed and either cleared for examination in toto or sectioned serially. The bursa was supplied by three pairs of extrinsic blood vessels. At 10 and 11 days of incubation, most intrinsic vessels were arranged in a superficial, hexagonal network. In regions of developing plicae, the hexagonal plexus extended into the core of each plica, forming middle plical vessels. The latter were interconnected across interplical areas by cross-connecting vessels. The middle plical vessels gave rise to small capillary offshoots, which soon increased in complexity, forming delicate loops. Branches extended from these loops through the subepithelial lamina propria to incipient epithelial buds by 12 days of incubation. All epithelial buds were supplied by at least one such branch, and similar branches extended to the basal aspect of the epithelium in areas where epithelial buds had not yet formed. This observation is consistent with the hypothesis that blood vessels induce formation of epithelial buds. At about 15 days of incubation, the cortex and medulla of each developing lymphatic follicle were defined clearly, and an intricate, web-like, capillary network coursed throughout the follicular cortex. The medulla appeared to be devoid of capillaries. The diameters of all intrinsic and extrinsic bursal blood vessels gradually increased throughout development. During post-hatching stages, the diameters of the extrinsic vessels continued to increase, whereas those of the intrinsic vessels were markedly decreased from late pre-hatching stages.     

Labelling of amoeboid microglial cells in rats of various ages following an intravenous injection of horseradish peroxidase

The macrophagic amoeboid microglial cells in the corpus callosum of postnatal rats were labelled following an intravenous injection of horseradish peroxidase (HRP). The earliest time when these cells were labelled was 3 h after the injection of HRP in postnatal (1-10 days) rats. Similar cells around the mesencephalic aqueduct and the fourth ventricle were also labelled. These cells, however, were weakly labelled in developing (11-20 days) and unlabelled in weaning (21-30 days) rats. The results suggest that in the postnatal rats, the HRP passed through the endothelial lining of the blood vessels and was then ingested by the amoeboid microglial cells. In the developing and older rats, the wall of blood vessels had developed fully thereby preventing the free passage of HRP into the brain tissues.     

Adaptive changes of rat liver cells induced by repeated intraperitoneal injections of d-galactosamine. III-Light- and electron-microscopic investigations of hepatocellular cytoplasmic changes.

In rats hepatocellular cytoplasmic changes after daily repeated D-galactosamine (GalN) intoxication--i.e. subacute GalN intoxication--were studied by light and electron microscopy. The number of GalN injections--and thus the days of survival--was between one and 30. The rats were killed six hours after the last GalN injection. Less degenerative changes were found after repeated GalN injections. An increased formation of atypical dense bodies (ADB), a temporary pronounced lipid accumulation and changes of the rough and smooth endoplasmic reticulum were prominent features of subacute GalN intoxication. The implications with respect to a modified GalN action in subacute GalN intoxication are discussed with special reference to biochemical data obtained in the same experimental model (Schuchhardt et al., 1977).     

Studies on retinal microangiopathy and coronary macroangiopathy in rats with streptozotocin-induced diabetes

Diabetes mellitus was induced in male rats by streptozotocin injection. After 4, 8 and 12 months, retinal and coronary vessels were prepared and their structure was studied. An age-dependent increase in thickness of the capillary basement membrane of retinal vessels was found both in controls and diabetics. Furthermore, diabetics had an added 20%, disease-related thickening, present after 4 months and thereafter. No significant change in coronary arteries was found. After 8 and 12 months, plasma lipids, triiodothyronin and thyroxin were measured. In diabetics, triglycerides were higher at both time intervals, high density lipoprotein was slightly increased and thyroxin was reduced after 8 months, and triiodothyronin was reduced after both 8 and 12 months. Diabetes of 12 months duration appears insufficient to induce significant arteriosclerotic changes in rats.     

Morphometric research on the structural changes in the liver of mice undergoing multiple exposures to a stress factor

C57BL/6 male mice were every day exposed to stressing influence. The liver was examined on day 4, 10, 21 after the start of the experiment and 3 days after the exposure was discontinued. These periods were characterized by increased concentration of ultrastructure membranes and intensified autophagocytosis. The volume changes in glycogen and autophagic vacuoles have been characterized by a strong correlation ratio. These changes were most prominent on day 10 of observation. The adaptive nature of changes is suggested.     

Age-related changes in the secretory pattern of FSH and LH in response to LH-RH in prepubertal female rats

Developmental changes in the pituitary responsiveness and the secretory pattern of FSH and LH in response to a single injection of LH-RH (100 ng/rat, s.c.) as estimated by increases in plasma concentrations of FSH and LH 10, 30 and 60 min after the injection were studied in female rats at 5, 10, 15, 20, 25 and 30 days of age. The pituitary responsiveness to LH-RH for both FSH and LH release increased from 5 to 15 days of age, reached a maximum on 15 days of age and declined thereafter, whereas a marked increase in the amount of these hormones in the pituitary occurred between 15 and 20 days of age. An apparent change in the secretory pattern of both FSH and LH was observed from 20 days of age onward. In groups up to 15 days of age, plasma concentrations of FSH and LH remained elevated 60 min after the injection of LH-RH, though the plasma concentration of these hormones returned to preinjection concentrations in groups at 20 days of age or later. These results indicate that the age-related changes in the secretory pattern of LH and FSH in response to LH-RH as well as changes in the pituitary responsiveness were apparent during the prepubertal period.     

Control of ovulation in cycling mares with ovarian steroids and prostaglandin

A combined progesterone-estradiol-17beta treatment was given in two experiments conducted to examine its effectiveness in controlling ovulation time in cycling mares. In the first experiment, the combined steroid (150 mg progesterone, 10 mg estradiol-17beta daily for 10 days) alone or combined with prostaglandin on the first and last days of steroid treatment resulted in ovulation in 15 of 16 mares 9-13 days after last injection, 13 of them on days 10-12. A CL present prior to treatment in one mare that received no prostaglandin persisted through and for 14 days after treatment. In the second experiment the combined steroid treatment started on the first or second day of estrus blocked ovulation in only 5 of 13 mares. Thus prostaglandin is necessary at least at the end of treatment. In both experiments a total of 20 mares with no luteal function at the end of steroid treatment ovulated on days 9-13 after last injection, 18 of these on days 10-12. These results indicate that the combined steroid-prostaglandin treatment can result in ovulations in a very restricted interval with apparently a normal distribution.     

A Comparison of the Oxytetracycline Preparations Aquacycline® and Terramycin®–100 with Regard to Absorption Characteristics,Local Tissue Reactions and Residues Following Dewlap Injections in Calves

In an experiment with 12 calves, Aquacycline® in a 5 % (OTC-A5) and a 10 % (OTC-A10) solution, was compared with Terramycin®-100 (OTC-C) by injecting 20 mg OTC/kg bwt. of these preparations in the dewlap and monitoring serum concentrations as well as tissue reactions and residues at the site of injection. All 3 preparations resulted in oxytetracycline (OTC) serum concentrations above 0.5 µg/ml of approximately 60 h. During this period, OTC-A5 resulted in a 39 % and OTC-A10 in a 20 % larger area under the serum concentration-time curve, as compared to OTC-C (P < 0.05). The recorded tissue reaction in the form of swelling during the first week following injection of OTC-A5 averaged 72 % of that after OTC-C (P < 0.01), while the mean swelling after OTC-A10 was 81 % of the corresponding value after OTC-C (P < 0.05). The OTC residue levels at the sites of injection were lower after OTC-A5, but none of the preparations resulted in OTC residues exceeding 0.3 mg at 28 days and about 0.15 mg at 42 days after injection. The pathological changes at the site of injection were somewhat more pronounced in those calves which received OTC-C. Accordingly, these results give some support to the claims that Aquacycline® offers advantages with respect to absorption characteristics and tissue tolerance.     

Additional effects of postpuberal estrogen injections on the vaginal epithelium in neonatally estrogenized mice

In the ovariectomized mice given 10 injections of 100 micrograms 17 beta-estradiol at intervals of 2 weeks from 60 days of age, the vaginal epithelium was atrophic when killed more than 2 months after the last injection. If mice given 3 daily injections of 20 micrograms 17 beta-estradiol from the day of birth were similarly treated with estradiol after postpuberal ovariectomy, the vaginal epithelium was stratified and hyperplastic at autopsy performed more than 2 months later. These changes in the epithelium persisted for at least 30 days after transplantation of the vaginae to normal ovariectomized hosts. Neonatal treatments only did not produce such persistent vaginal changes. In view of these results, additional effects of neonatal and postpuberal injections of estrogen on the vaginal epithelium are evident. However, effects of such neonatal and postpuberal injections of estrogen might be transient on the uterine epithelium, since abnormal proliferation was not observed in it.     

Localization of neuropeptide Y and norepinephrine in the porcine ovarian artery and their influence on the local blood pressure

The aim of the present study was to determine: (i) the presence of dopamine-beta-hydroxylase (DbetaH)- and neuropeptide Y (NPY)-immunoreactive (IR) nerve fibres in the wall of the porcine ovarian artery, (ii) the influence of NPY and norepinephrine (NE) on the contractile activity of the pig ovarian arteries, and (iii) the pharmacological analysis of the interaction between NPY and NE in the isolated porcine ovarian arteries collected from immature pigs and from animals in different days of the estrous cycle. Ovarian arteries for immunohistochemistry and isolated arteries for pharmacological studies were excised from immature pigs and mature animals on days 1-5, 8-13 and 17-20 of the estrous cycle. The study showed that both DbetaH- and NPY-IR nerve fibres were present in the pig ovarian arteries in all periods examined, and, that in some fibres DbetaH and NPY were co-localized. Both NE (10(-6) M) and NPY (10(-7) M) increased blood pressure of examined preparations, however, NE caused stronger changes in the vessel wall tension (P<0.001), than did NPY. NE significantly increased (P<0.001) blood pressure of all isolated arteries, however, this response was stronger in vessels from days 1-5 of the cycle, when compared to days 8-13 (P<0.01), 17-20 and immature pigs (P<0.001). NPY increased significantly blood pressure in isolated arteries from days 8-13 and 17-20 of the cycle (P<0.001), while in preparations taken from immature pigs and animals on days 1-5 of the estrous cycle this response was somewhat weaker (P<0.01). A higher elevation (P<0.001) of blood pressure after NPY administration was observed in isolated arteries from days 17-20 of the cycle, when compared to vessels from days 1-5 and 8-13 and those from immature pigs. Moreover, NE significantly intensified (P<0.001) an increase in the blood pressure in isolated arteries pre-treated with NPY in all periods examined. NPY insignificantly (P>0.05) potentiated increase of blood pressure in NE pre-treated vessels of immature pigs and in isolated arteries from days 17-20 of the cycle and significantly (P<0.05) in vessels from days 1-5 and 8-13 of the estrous cycle. Our results indicate that DbetaH- and NPY-IR nerve fibres are present in the pig ovarian arteries. NE and NPY administered alone increased blood pressure in the pig isolated ovarian artery and simultaneous administration of both substances caused each other potentiation of vasocontractile effect, however, the strength of observed changes was dependent on the stage of the estrous cycle.     

Response of ovaries and serum steroid levels after treatment of cystic ovarian disease in dairy cattle I. treatment with corticosteroids and a combination of human chorionic gonadotropin and progesterone

Of 38 cows with ovarian follicular cysts (cysts), 10 were injected intramuscularly with 20 mg of betamethasone or 10 mg of dexamethasone (CC) and 28 intravenously with a combination of 3,000 IU of human chorionic gonadotropin and 125 mg of progesterone (HCG·P). Ovarian responses and changes in serum levels of sex steroids were examined after both of the treatments. Percent of the cows in which cysts luteinized 10 days after the treatment in the HCG·P-treated cows was significantly higher (P<=0.05) than in the CC-treated ones. Serum levels of progesterone and estrogens (estrone and estradiol-17β) in the cows were not affected 10 days after CC injection. On the contrary, serum progesterone increased significantly (P<=0.05) and serum estradiol-17β decreased significantly (P<=0.05) 10 days after HCG·P injection. Serum progesterone did not respond to HCG·P treatment in the cows previously treated unsuccessfully with gonadotropin, while progesterone increased significantly (P<=0.05) in response to HCG·P treatment in those given no treatment before.     

Fertilizing competency of multiple ovulated eggs in the domestic fowl (Gallus domesticus)

Fertilizing competency of multiple ovulated eggs in the domestic fowl was examined by fertilization in vitro and early development in culture. Normal laying hens (White Leghorn) were treated with 75 IU of PMSG for 7 days followed by injection of anterior pituitary extracts from chickens (CAPE). Ovulation began to occur 7.5 h after injection of CAPE. These hens ovulated 1-7 ova but some premature ovulation of GV stage ova were observed. In vitro fertilization of the multiple ovulated ova was examined by inseminating 10(6)-10(7) sperm onto the germinal disks in m-Ringer's solution. The gamete or zygote nuclei were detected by DNA specific fluorescence using DAPI (4',6'-diamidino-2-phenylindole) in the histological section prepared from the germinal disk. Process of fertilization was examined in the eggs incubated for 4 h after insemination in DMEM + liquid albumen at 41 degrees C under the atmosphere of 5% CO2 in air. Fertilization rate of the total multiple ovulated eggs was 55% (11/20), in which 90% (9/10) and 10% (1/10) in the eggs recovered 7.5-8.5 h and 9.0-9.5 h after CAPE injection were obtained, respectively. Normal pronuclei were formed in five eggs of those recovered 7.5-8.5 h after CAPE injection. Early development after fertilization in vitro was also examined by incubation for 12 h in DMEM + liquid albumen at 41 degrees C under the atmosphere of 5% CO2 in air. Although development in vitro was delayed compared to that in utero condition, normal development was observed in naturally and multiple ovulated eggs.(ABSTRACT TRUNCATED AT 250 WORDS)     

Early effects of ionizing radiation on the microvascular networks in normal tissue

Microvascular networks, which control the delivery of oxygen and nutrients and the removal of metabolic waste, are the most sensitive part of the vascular system to ionizing radiation. Structural and functional changes in microvascular networks were studied in locally irradiated (single 10-Gy dose) hamster cremaster muscles observed 3, 7 and 30 days post-irradiation. Networks were selected in reference to a well-defined location in the tissue to reduce heterogeneity due to spatial variations. Intravital microscopy was used to measure structural and functional parameters in vivo. A factorial design was used to examine the effects of radiation status, time postirradiation, and network vessel type on the structure and function of microvascular networks. While the diameter of microvessels in control animals increased significantly with age, vessel diameter in irradiated vessels decreased significantly with age. Red blood cell velocity in irradiated networks at 3 and 30 days postirradiation was significantly lower than in control networks. There was a significant decrease in capillary surface area and a significant increase in vessel hematocrit in irradiated animals. Blood flow in irradiated vessels was significantly lower than in control vessels. Changes in functional parameters were evident at 3 days postirradiation while changes in structural parameters occurred later. All vessel types were not damaged equally by radiation at every time examined.     

Capillary plexus development in the day five to day six chick chorioallantoic membrane is inhibited by cytochalasin D and suramin

The chick chorioallantoic membrane (CAM) is a valuable model for evaluating angiogenesis and vasculogenesis. Our purpose was to characterize the formation of the CAM vasculature, in particular the capillary plexus, between days five and six after fertilization and to examine the mode of action of cytochalasin D and suramin on vascular development during this interval. The CAM increased 20-fold in size between days five and six, during which time the capillary plexus forms by both migration of mesodermal blood vessels toward the ectoderm and by the formation of new vessels from angioblasts near the ectoderm. Between days five and six, the CAM becomes thinner, and the density of the mesodermal cells decreases. To determine the mode of action of anti-angiogenic drugs on the day five to day six CAM, various concentrations of cytochalasin D or suramin were added directly to day five CAMs, and their effects were evaluated on day six. Both drugs significantly inhibited CAM growth, altered branching patterns of the major vessels, decreased area of the major vessels, and inhibited the formation of the capillary plexus by inhibiting both vasculogenesis and the migration of mesodermal blood vessels to the ectoderm. Cytochalasin D also inhibited compartmentalization of the plexus. Cytochalasin D and suramin were inhibitory at similar doses. This study provides new information on early CAM development, establishes the mode of action and dose dependency of cytochalasin D and suramin on day five to day six CAMs, and demonstrates that the day five to day six CAM provides a useful assay to examine the effect of anti-angiogenic drugs on blood vessel development, including capillary plexus formation.     

Use of hepatocytes cultured in microcarriers for correction of acute hepatic insufficiency

The ability of hepatocytes cultured on Biosilon microcarriers to secrete albumin and to conjugate bilirubin were examined for the purpose of developing an artificial liver support system. Cultured hepatocytes were able to synthesize 100-120 micrograms albumin per 10(5) cells in 24 hours and to conjugate about 20 micrograms/hr of bilirubin for at least 5 days. Rats with liver failure caused by i.p. injection of CCL4 or D-galactosamine were subjected to hemosorption via minicolumns containing 2 ml of Biosilon microcarriers with 40 X 10(6) cultured hepatocytes. The procedure was performed 20-24 h after hepatotoxins injection and lasted for 3 h at a flow rate of 60 ml/h. This reduced mortality from 100% to 20% after 48% and to 40% after 7 days in case of CCl4 and from 100% to 40% after 48 h and 7 days in case of D-galactosamine. Our results suggest that hepatocytes cultured on microcarriers may be efficiently applied to correction of fulminant hepatic failure caused by different hepatotoxins.