The human carotid atherosclerotic plaque stimulates angiogenesis on the chick chorioallantoic membrane.

The chick chorioallantoic membrane was used to determine whether the carotid atherosclerotic plaque stimulates angiogenesis. Carotid endarterectomy specimens (1 mm3) with fibromuscular plaque (n = 8) and complicated plaque (n = 11) were implanted on the membrane on day nine of incubation and the response evaluated on day 11. Following fixation in situ with 10% formalin the angiogenic response was evaluated by: (1) examining whole membrane mounts, (2) quantitatively from a vascular density index and (3) from a histological study. Unmanipulated chorioallantoic membrane (n = 11) and plaque boiled prior to implantation (n = 6) served as controls. The vascularity of whole mounts of both fibromuscular and complicated plaque was greater than the controls. Vessel density of the membrane was estimated by counting the number of vessels intersecting four concentric circles (144.5 mm total circumference) placed on the formalin fixed membrane. The vascular density index due to the fibromuscular plaque (390.6 +/- 8.3) and complicated plaque (391.0 +/- 14.9) were similar (P greater than 0.9) but were significantly greater (P less than 0.001) than the unmanipulated membrane (327.9 +/- 5.6) or after treatment with the boiled plaque (283.8 +/- 15.6). Transforming growth factor beta 1 confirmed the validity of the experimental model to study angiogenesis. The histology of the chorioallantoic membrane due to either type of plaque was similar. Numerous vessels surrounded the plaque, and intraplaque vessels containing nucleated chick erythrocytes were observed. Although scattered vessels surrounded the boiled plaque, intraplaque vessels were not observed. This study demonstrates that the atherosclerotic plaque has angiogenic properties that may account for the increase in vasa vasorum that is associated with the plaque.

Adipose tissue determinations in cadavers--a comparison between cross-sectional planimetry and computed tomography.

The adipose tissue surfaces in 11 slices (+/- 5 cm from the umbilicus) were compared in two cadavers using computed tomography (CT) versus planimetry of band-sawed slices of the corresponding sections. A very close correlation was found with partial correlations of around 0.90. Retroperitoneal fat formed a considerable proportion of the total adipose tissue surface in the slices. The results were similar whether fat was defined as -250 to -50, -190 to -30, or -140 to -40 Hounsfield units. These data indicate that CT measurements agree closely with a direct morphometric method and thus can be used as a 'gold standard' for future development. The fact that fat which is located extraperitoneally, but still intraabdominally, constitutes a significant proportion of the slice surface in the umbilical region indicates that data relating intraabdominal fat measurements to metabolic functions must be interpreted with caution.

The origin and distribution of vasa vasorum at the bifurcation of the common carotid artery with atherosclerosis.

The purpose of our study was to determine the origin and relation of vasa vasorum to atherosclerotic plaque at the bifurcation of the common carotid artery. We randomly selected 12 unembalmed adult human cadavers, 40-96 years of age. We prepared luminal casts of the arteries from eight cadavers and cleared the arteries from the remaining four cadavers. A network of vasa vasorum surrounding atherosclerotic plaque was observed in five luminal casts and in two cleared specimens; the vasa vasorum originated from the superior thyroid and ascending pharyngeal arteries. Three of the five luminal casts also demonstrated vasa vasorum arising directly from the internal carotid artery distal to the plaque. An extensive network of vasa vasorum was not observed in specimens from the five cadavers relatively free of gross atherosclerotic plaque. Our findings demonstrate the importance of the external carotid artery in giving rise to the vasa vasorum that supply the areas of atherosclerotic plaque.

Effects of shock waves on the rat ovary.

The studies reported in this paper were undertaken to investigate the effect of shock waves on ovarian function. In an initial study, five female Wistar rats underwent general anesthesia. One ovary was then exposed to 1500 shock waves at 20 kV using the Dornier XL-1 experimental lithotripter. One week following this treatment, the animals were sacrificed and both ovaries were step sectioned. We counted all follicles that contained both an oocyte and antrum. These follicles in turn were classified further as being healthy or atretic. No statistical difference in these counts was observed between shocked and unshocked ovaries taken from the same animal. In the definitive study, thirty-one adult female Wistar rats underwent unilateral oophorectomy. These rats were then divided into treated, sham-treated and control groups. The eleven treated animals were given general anesthesia and received 1500 shocks at 15 kV directed at their remaining ovary using the Dornier XL-1 lithotripter. The ten sham treated animals received identical anesthesia and duration of waterbath immersion as the treated group. In contrast, however, sham treated animals were positioned outside the path of the focused shock wave. The control group received no treatment. Normal estrus cycle was maintained in all groups following shock wave exposure. Two weeks after this exposure, all three groups of animals were allowed to mate. All animals in each of the three groups became pregnant. There was no statistical difference in litter size or fetal weights at three weeks gestation. No gross teratogenic effects were observed. The rat ovary appears to be structurally and functionally resistant to the shock wave energy levels used in this experiment.

Effect of chronic alcohol consumption on the pregnant mare serum gonadotropin-induced luteinizing hormone surge.

The studies reported in this paper were undertaken to investigate the effect of chronic (10 day) alcohol consumption on female pituitary-gonadal function. The immature female rat model treated with pregnant mare serum gonadotropin (PMSG) was used since it results in a highly reproducible luteinizing hormone (LH) surge and ovulation. Twenty-day-old female rats were placed on diets that were either (1) unrestricted (ad libitum); (2) contained 5% ethanol in a liquid diet, or (3) isocalorically pair-fed with the liquid diet to the ethanol group. After 10 days on their respective diets, the groups were subdivided and given either 8 IU of PMSG in 0.1 ml saline or 0.1 ml saline s.c. between 10.00 and 11.00 h. The animals were sacrificed by decapitation at 24, 48, 52, 54, 56, 58, 60 and 62 h after injection. Trunk blood was obtained for serum measurements of LH. The uteri were weighed and prepared for the histological study. In all dietary groups, serum LH levels were significantly higher in the PMSG-treated animals when compared to the saline controls at all time intervals with the exception of the alcohol 58-hour group. In the ad libitum animals, plasma LH concentrations were highest at 52 h following hormone administration. The serum LH concentrations were highest at 56 h after hormone administration in the pair-fed group and were significantly less than the ad libitum group at 52, 54, 56, and 58 h after PMSG stimulation. No significant plasma LH surge was observed in the alcohol group and the LH concentrations were significantly less than the pair-fed rats at 56 and 58 h after PMSG treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Ethanol inhibition of compensatory ovarian hypertrophy in unilaterally ovariectomized rats.

The purpose of this study was to determine the effect of ethanol consumption on compensatory ovarian hypertrophy (COH) which occurs following unilateral ovariectomy. Holtzman rats, 40 days old, were either unilaterally ovariectomized or sham-ovariectomized. The rats were then placed into subgroups which would receive either an ad libitum chow and water diet, a liquid diet, or a liquid diet containing 5% ethanol. The animals were maintained on their respective diets for 11 days. The rats were killed at 51 days of age, and the ovaries and uteri were removed, weighed, and prepared for histological investigation. The results showed that uteri from ethanol-fed animals failed to develop epithelial glands and exhibited a condensed stroma in comparison to uteri from animals fed ad libitum or pair-fed to ethanol-fed rats. Also, rats that were fed ad libitum had a COH of 82 +/- 16% and rats that were pair-fed a liquid diet had 114 +/- 28% COH; rats that were fed the liquid diet containing ethanol did not experience compensatory ovarian hypertrophy (-3 +/- 10%). Histologically, the ovaries of rats fed ad libitum showed large numbers of corpora lutea and only a few mature ova. The histology of ovaries from pair-fed animals was similar to those from animals fed ad libitum. In contrast, the ovaries from the animals fed the ethanol diet had nearly twice as many mature ova but only one-fourth as many corpora lutea as the number seen in ovaries from the other groups. The data show that ethanol consumption inhibits COH by suppressing ovulation and the subsequent luteal formation.

Estrous cyclicity in rats fed an ethanol diet for four months.

This study is designed for better understanding the effect of chronic ethanol treatment on ovarian function in sexually mature rats by assessing the estrous cycle. Rats were fed the following diets for 17 weeks: a liquid diet containing 5% (w/v) ethanol, a liquid diet without ethanol (pair-fed) or laboratory chow and water ad lib. Estrous cycles were determined throughout the 17 weeks and the rats were necropsied at proestrus and metestrus. Ethanol-fed rats had significantly more irregular estrous cycles than did controls, and the duration of an estrous cycle in the ethanol-treated rats was statistically longer than that of controls. However, ovarian and uterine weights and ovarian and vaginal morphology of ethanol-fed rats were similar to those of controls.

Effects of ethanol on superovulation in the immature rat following pregnant mare's serum gonadotropin (PMSG) or PMSG and human chorionic gonadotropin treatment.

We sought to determine whether superovulation could occur in immature rats on a 5% ethanol diet and treated with pregnant mare's serum gonadotropin (PMSG) alone or with human chorionic gonadotropin (hCG). Holtzman female rats were divided into five groups at 20 days of age. Six rats (Group I) were killed at that age. Ten rats (Group II) were placed on an ad libitum laboratory chow diet and killed on Day 33. Twenty-four rats (Group III) were placed on an ad libitum laboratory chow diet. Twenty-four rats (Group IV) were placed on 5% ethanol liquid diet, while 24 rats in Group V were pair-fed with the animals in Group IV. At 30 days of age, 12 rats from each Group, III, IV, and V, received 25 IU of PMSG s.c. and were killed 74-76 h later. The remaining 12 rats from each Group, III, IV and V, received 25 IU of PMSG and 54-56 h later received 10 IU of hCG and were killed 20 h later. Ovulation occurred in all the rats of Groups III and V that received PMSG alone or with hCG. In the ethanol-treated rats that received PMSG alone, 75% ovulated, while 92% ovulated that received PMSG and hCG. The number of ova shed in the ethanol-PMSG-treated rats was significantly less than in the ethanol-PMSG-hCG-treated animals and in the controls. The uterine weights and morphology of the animals in Group IV were similar to those in Groups III and V. The study indicates that ethanol does not have a direct gonadotoxic effect on the ovary but indicates that ethanol has an effect on the hypothalamus and/or the pituitary, thereby disrupting the synthesis and/or release of luteinizing hormone releasing hormone (LHRH) or luteinizing hormone (LH).

Female reproduction during chronic ethanol consumption in rats.

This study was undertaken to ascertain ovarian function under the conditions of ethanol withdrawal and continued ethanol treatment to distinguish between a temporary delay in ovarian activity and a permanent suppression of ovarian function. Immature rats were fed the following diets for 16 weeks: a liquid diet containing 5% ethanol, a liquid diet without ethanol (pair-fed controls), a liquid diet with 5% ethanol for eight weeks followed by laboratory chow and water for eight weeks, or chow and water ad lib. Vaginal patency was significantly delayed in both groups of ethanol-treated rats compared to controls. The duration of an estrous cycle for the rats in the ad lib group was 5.0 +/- 0.3 days, while a "regular" estrous cycle was four to six days in duration. The rats which received ethanol for 16 weeks exhibited more irregular estrous cycles (both less than 4 and greater than 6 days) than the rats with other treatments and the cycles were significantly longer. After 16 weeks of treatment, the rats were mated; ethanol was not given during pregnancy. The average number of pups per litter and body weight of the offspring were similar for all groups. These data show that although ethanol alters normal cyclic activity, it does not totally suppress ovarian function since alcohol-treated rats were capable of mating and delivering viable offspring.

Ethanol-induced alterations in the morphology and function of the rat ovary.

The purpose of this study was to compare the effects of different dosages of ethanol on ovarian morphology and function. Holtzman rats, 20 days old, were divided into groups as follows: The rats in Group I were autopsied at 20 days of age, and those in Group II were placed on ad libitum chow and water diet; the rats in Groups III and V were fed on a liquid diet containing 2.5% or 5% ethanol respectively; Groups IV and VI were pair-fed controls to Groups III and V, respectively. Rats in Groups II, III, IV, and VI were maintained on the diets for 50-55 days and killed at late proestrus-estrus, while the animals in Group V did not exhibit estrous cycles and were killed on day 55 of treatment. The average increase in body weights of rats in Groups II, III, and IV was significantly greater than the increase in body weights of rats given 5% ethanol or their pair-fed controls. In the rats treated with 5% ethanol, vaginal opening was significantly delayed from the controls, estrous cycles were absent, ovarian weights were similar to those of the 20-day-old rats, ovaries contained corpora lutea of only one estrus, uteri weighted less than controls, and histologically, the uteri and vaginae were similar to those of 20-day-old rats. However, in the rats treated with 2.5% ethanol, all of the parameters were similar to those of the controls. The average serum alcohol level for the rats on the 5% ethanol diet was 249 mg%; the serum alcohol levels were at the lower limit of detection for the rats on the 2.5% ethanol diet. The data show that ovarian function was suppressed in the rats that received the 5% ethanol but not in rats on the 2.5% ethanol diet.

The utilization of xeroradiography for radiography of cross-section of thin cadaveric slices.

Although xeroradiography has had many medical applications, its utilization and the advantages it offers in the study of cadaveric sections has not previously been fully explored. It was our purpose to determine possible advantages or disadvantages this modality might have for this purpose. The edge enhancement which xeroradiography allows is a distinct advantage since the various anatomic structures become more clearly defined. On the other hand, the hollow viscera and the lumina of blood vessels are not enhanced, but rather blend with the walls of these "hollow" structures. Also, there is some loss of detail in the depiction of some of the glandular structures, such as the seminal vesicles. An artist's assistance to define these by comparison with the gross specimen gives us an optimum result. An x-ray enhanced image is thereby attained which exceeds the usual line drawing or artist's depiction of the cross-sectional specimen, and both can be similarly labeled for study.

The effect of ligation or separation between the intrauterine device horn and adjacent ovary on implantation in the hamster.

The purpose of this study was to determine whether the luteolytic action of an intrauterine device (IUD) is suppressed following interruption of the continuity between the IUD uterine horn and the adjacent ovary. After several estrous cycles, a silk IUD was placed in the cervical end of one uterine horn of adult hamsters. The animals were then mated, and on day 6 of gestation the ovary and oviduct contralateral to the IUD were removed and a ligature was placed between the IUD horn and the adjacent ovary. The hamsters were killed on day 9 or day 13 of gestation. Fetal development in the contralateral horn was suppressed on day 13 but not on day 9. In the second study the animals were treated as in the first study except that the communication between the IUD horn and the adjacent ovary was severed completely. The hamsters were killed on day 9 or day 13. On both days 9 and 13 normal fetal development was observed in the control horn; no implantation sites were present in the IUD side. In the control (non-IUD) animals of each study, normal fetuses were present in both uterine horns. The study demonstrates that luteolysis does not occur if there is complete disruption of the communication between the IUD horn and ovary. The study also demonstrates that, since implantation did not occur in the IUD horn with a normally functioning ipsilateral ovary, the luteolytic action of the device is not the prime factor in suppressing implantation in the hamster.

The interaction of clomiphene, estradiol, and progesterone in the control of rat uterine glycogen metabolism.

Uterine glycogen accumulation was studied in ovariectomized rats treated with all combinations of clomiphene citrate (0.25 mg/kg) estradiol (1.0 micron g) and progesterone (5.0 mg). The rats were given three consecutive daily dosages and killed 24 hours after the final dosage. Based on biochemical data, either estradiol or clomiphene increased uterine glycogen concentration and total glycogen, but progesterone did not. Progesterone significantly suppressed both the estradiol and clomiphene-induced glycogen increases. Based on the histochemical results, progesterone also suppressed the estradiol and clomiphene-induced glycogen responses, but the tissue affected differed. Clomiphene markedly increased luminal epithelial glycogen whereas estradiol induced primarily myometrial glycogenesis. Progesterone completely suppressed the clomiphene-induced epithelial effect and partially suppressed the estradiol-induced myometrial effect. Clomiphene also suppressed the estradiol-induced myometrial response. The results indicate that progesterone does have a significant interaction with clomiphene in the control of uterine morphology and biochemistry. The results also stress the importance of correlated histochemical and biochemical studies in the study of clomiphene-induced uterine glycognesis.

PIP: The interaction of clomiphene citrate (CL), estradiol-17 (E2), and progesterone (P) in the control of rat uterine glycogen metabolism was studied by histochemical and biochemical techniques. Bilaterally ovariectomized rats were given CL (.25 mg/kg), E2 dipropionate (1.0 mcg/rat), or P (5.0 mg/rat) alone or in combination with 1 or more. Rats were treated for 3 days. Uterine horns were examined and analyzed for glycogen levels. E2 and CL alone increased uterine glycogen concentration and total glycogen while P was ineffective. P, however, significantly suppressed E2- and CL-induced glycogen increases (p .05). P partially suppressed E2 induced myometrial response and completely suppressed CL epithelial response. Results indicate an important interaction between P and CL in the control of uterine morphology and biochemistry. It is recommended that histochemical and biochemical analysis of CL-induced uterine glycolysis be considered in any study.

Effect of direct injection of neutrophils from uterine horns of rats containing intrauterine devices into recipient pregnant uteri.

The purpose of this study was to determine whether or not neutrophils from uterine horns containing an intruterine device (IUD) inhibit implantation of rat blastocysts. On day 4 (6:00 P.M. to 8:00 P.M.) of gestation the uterine horns of the recipient rats were exposed and injected with neutrophils, supernatant luminal fluid, or Hanks' solution. The total volume injected was 30 mul. All of the rats were killed on day 12 of pregnancy, and the number of fetuses and resorption sites was determined. The data show that, in rats injected with neutrophils on day 4 of gestation, implantation was significantly increased as compared with controls injected with Hanks' (vehicle) solution. Whether the supernatant fluid had an effect in suppression of implantation could not be determined, since the effect was not statistically different from that observed in either the vehicle- or neutrophil-injected groups. However, the responses observed with the supernatant-injected groups. However, the responses observed with the supernatant-injected group were intermediate between those of the control horns and those of the neutrophil-injected horns. The finding supports the concept that in IUD-bearing animals in which there is a rapid influx of neutrophils the cells play a major role in suppressing implantation.

Influence of the IUD on ovarian activitiy in the hamster and rat.

PIP: The efects of an unilaterally placed IUD on the maintenance of pregnancy in the contralateral horn was studied in rats and hamsters. Implantation and the number of corpora lutea were clearly affected by the IUD, but there was no effect on the number of eggs shed. There were no fetuses found in either horn in hamsters receiving a unilateral silk IUD. However, in rats, fetuses were found in the contralateral horn, though not in the IUD-bearing horn. Thus, the IUD has a different effect on ovarian function in rats and hamsters. The results suggest that the IUD causes regression of the corpus luteum in hamsters, and alters the uterine environment in rats.^ieng

A circadian rhythm of mitotic activity in the uterine luminal epithelium of the rat: effect of estrogen.

The study was designed to determine whether or not the rat uterine luminal epithelium exhibits a mitotic circadian rhythm and to ascertain the effect of estrogen treatment at different time periods on the uterine epithelial mitotic response. Immature rats were injected with either sesame oil (controls) or 60 ng of estradiol-17 beta at eight time periods and were necropsied 24 hours after treatment. Colchicine was administered IP two hours before autopsy. Peak mitotic activity was observed during the nocturnal phase (0300) for both the control and estrogen-treated rats. The nadirs were recorded during the diumal phase (1800 and 1200 for the control and estrogen groups, respectively). The differences between low and high values were 1100% for the control rhythm and 101% for the estrogen animals. The data demonstrate the existence of overt circadian rhythms in the uterine epithelium for both control and estrogentreated rats.