The multinomial logistic regression model for predicting the discharge status after liver transplantation: estimation and diagnostics analysis.

The multinomial logistic regression model (MLRM) can be interpreted as a natural extension of the binomial model with logit link function to situations where the response variable can have three or more possible outcomes. In addition, when the categories of the response variable are nominal, the MLRM can be expressed in terms of two or more logistic models and analyzed in both frequentist and Bayesian approaches. However, few discussions about post modeling in categorical data models are found in the literature, and they mainly use Bayesian inference. The objective of this work is to present classic and Bayesian diagnostic measures for categorical data models. These measures are applied to a dataset (status) of patients undergoing kidney transplantation.

Regulatory role of inhibin in follicle-stimulating hormone secretion and folliculogenesis in the guinea pig.

The effects of unilateral and bilateral ovariectomy and passive immunization against inhibin on follicle-stimulating hormone (FSH) secretions and follicular development in the guinea pig were investigated. Bilateral ovariectomy decreased plasma immunoreactive (ir-) inhibin rapidly and increased plasma FSH significantly. Unilateral ovariectomy decreased plasma ir-inhibin and increased plasma FSH temporarily, and doubled the number of ova released from the remaining ovary at the subsequent ovulation in guinea pigs. Injection of 1.0 ml inhibin antiserum significantly increased concentrations of plasma FSH at 6 hr onwards and the number of small follicles (100-200 microm in diameter) at 48 hr after the injection in guinea pigs bearing progesterone-containing implants. In vitro bioassay showed that inhibin antiserum could neutralize the suppression of ovarian homogenate on FSH secretion from cultured rat anterior pituitary cells. These results confirm the evidence that the ovary is the main source of inhibin secretion and both in vitro bioassay and passive immunization against inhibin show that the inhibin is a major regulator in the follicular development through FSH secretion in guinea pigs.

Regulation of follicle-stimulating hormone secretion by estradiol and dimeric inhibins in the infantile female rat.

Plasma and ovarian levels of the dimeric forms of inhibin and plasma estradiol-17beta were investigated and compared with changes in plasma gonadotropins from Postnatal Day (PND) 5 to PND 30 in the female rat. The inhibin subunit proteins were localized in follicular granulosa cells of the ovary. Plasma immunoreactive inhibin levels were low until PND 15 and increased thereafter. Plasma levels of inhibin B (alpha and beta(B) subunits) remained very low until PND 15 and then increased by approximately 24-fold. In contrast, plasma levels of inhibin A (alpha and beta(A) subunits) were relatively low and steady until PND 20, then increased by approximately 3-fold at PND 25. Changes in ovarian inhibin A and B levels closely resembled those in plasma levels. Plasma FSH levels were low at PND 10 but started to peak from PND 15 and remained high until PND 20, followed by a remarkable reduction at PNDs 25 and 30. This dramatic fall in FSH coincided with the rise of inhibin A. A significant inverse correlation was observed between plasma FSH and plasma inhibin A (r = -0.67, P < 0.0002), ovarian inhibin A (r = -0.48, P < 0.01), plasma inhibin B (r = -0.48, P < 0.05), and ovarian inhibin B (r = -0.54, P < 0.01). Plasma estradiol-17beta levels were elevated from PND 5 through PND 15, then fell sharply through PND 30. Plasma estradiol-17beta was significantly and positively (r = 0.75, P < 0.0002) correlated with plasma FSH. Plasma LH rose to higher levels at PND 15 and tended to be lower thereafter. The inhibin alpha, beta(A), and beta(B) subunits were localized to primary, secondary, and antral and large antral follicles, but the types of these immunopositive follicles varied with age. It appeared that, at PND 25 and afterward, all three subunits were mainly confined to large antral follicles in the ovary. We conclude that estradiol-17beta likely is the major candidate in stimulation of FSH secretion in the infantile female rat. We also conclude that inhibin regulation of pituitary FSH secretion through its negative feedback in the infantile female rat begins to operate after PND 20. We suggest that this negative feedback is achieved by increases in plasma levels of the two dimeric forms, and that inhibin A appears to be the major physiological regulator of FSH secretion at the initiation of this mechanism. We also conclude that large antral follicles in the ovary are the primary source of these bioactive inhibins that are secreted in large amounts into the circulation after PND 20.

Effects of diesel exhaust particles on blood pressure in rats.

The effects of diesel exhaust particles (DEP) on pulmonary functions and consequent diseases are well known, but there have been few reports concerning involvement of the cardiovascular system. In order to assess a direct action of DEP on cardiac tissue, the effects on blood pressure of intravenous administration of 12 or 120 mg/kg DEP to anesthetized rats were studied for a 15-min period. DEP (120 mg/kg) significantly lowered blood pressure for 25 s with no signs of arrhythmia or mortality, a phenomenon seen in guinea pigs. After 25 s blood pressure gradually returned to control levels and was maintained for 15 min. The 12-mg/kg DEP concentration did not markedly affect rat blood pressure. Pretreatment with atropine (24 mg/kg) blocked the DEP-induced fall in blood pressure, while pretreatment with propranolol (48 mg/kg) proved ineffective against DEP, suggesting involvement of the parasympathetic system. Data show that the rat is less sensitive to DEP-induced effects on blood pressure and may be a poor model to reflect cardiovascular changes.

Diesel exhaust-induced airway hyperresponsiveness in c-Ha-ras transgenic mice.

Recently the quantity of diesel exhaust (DE) emissions, which contain a variety of chemicals and can induce pulmonary carcinoma in animals, has been increasing in Japan. To assess the toxicity of DE, we evaluated airway hyperresponsiveness after exposure to DE in the rasH2 (CB6F1-TgHras2) mouse, which carries c-Ha-ras genes and shows marked sensitivity to treatment with various genotoxic carcinogens such as methylnitrosourea and dimethylbenzanthracene. We exposed rasH2 mice (n=18) and their nontransgenic littermates (n=19) to room air or 3 mg/m(3) DE for 4 weeks, measured their respiratory resistance (Rrs) during inhalation of acetylcholine (ACh; 0.005, 0.01, 0.02, 0.04, 0.08, 0.16, 0.31, 0.63, 1.28, 2.5, 5, or 10 mg/ml) for 2 min, and calculated the provocative ACh concentration needed to cause a 50% increase (PC(150)) in Rrs. At all doses of ACh, Rrs was significantly higher (P<0.05) in rasH2 mice exposed to DE than in those exposed to room air. In addition, Rrs in the DE-exposed rasH2 animals was significantly higher (P<0.05) at 0.16, 0.31, and 0.63 mg/ml ACh than in DE-exposed nontransgenic littermates. The PC(150) (mean+/-standard error) of DE-exposed rasH2 mice was 3.4+/-1.9 mg/ml, that in rasH2 mice exposed to room air was 10.6+/-2.5 mg/ml, and that in DE-exposed nontransgenic animals was 10.9+/-3.7 mg/ml. In conclusion, DE causes airway hyperresponsiveness in rasH2 mice and may induce the expression of c-Ha-ras genes.

Diesel exhaust (DE) affects the regulation of testicular function in male Fischer 344 rats.

To investigate the effects of diesel exhaust (DE) particles on the reproductive system, male Fischer 344 rats at 13 mo of age were exposed to clean air or DE at particle concentrations of 0.3, 1, or 3 mg/m3 for 8 mo. DE did not markedly affect testicular and body weights. However, DE at 0.3 mg/m3 significantly decreased prostate and coagulating gland weights, accompanied by a reduction in thymus and adrenal gland weight. In contrast, there was a significant rise in the weights of prostate, seminal vesicles, and coagulating glands in the 3 mg/m3 DE group. In rats exposed to 0.3 or 1 mg/m3 DE, serum luteinizing hormone (LH) and testosterone increased significantly, while a rise in testicular testosterone was noted with 3 mg/m3 DE. The concentrations of follicle-stimulating hormone (FSH) and inhibin as well as the sperm head counts were not markedly altered in any treatment group. Positive staining with inhibin-alpha subunit and 3beta-hydroxysteroid dehydrogenase (3beta-HSD) were observed in Sertoli cells and Leydig cells, respectively. Immunolocalization of inhibin-alpha subunit and 3beta-HSD was not changed by exposure to DE. In conclusion, DE appears to exert greater effects on accessory glands than on testes in Fischer 344 rats, and the responsiveness of rats is less than that found in mice.

Exposure of neonatal female rats to p-tert-octylphenol disrupts afternoon surges of luteinizing hormone, follicle-stimulating hormone and prolactin secretion, and interferes with sexual receptive behavior in adulthood.

The present study investigated the effects of exposure of neonatal female rats to p-tert-octylphenol (OP) on estrogen-induced afternoon surges of LH, FSH, and prolactin (PRL) secretion, and on sexual behavior in adulthood. After birth, one group of female Wistar rat pups received s.c. injections of OP (100 mg/kg body weight [BW]; OP group) dissolved in DMSO, while the control group received DMSO only (DMSO group). In order to make a qualitative comparison, a third group was injected with estradiol-17beta (500 microg/kg BW; estradiol group) dissolved in DMSO. Injections were given on Days 1, 3, 5, 7, 9, 11, 13, and 15 of age. The rats from the OP and estradiol groups that were used for subsequent experiments were in persistent vaginal estrus. Spontaneous LH surge measured at Postnatal Days (PND) 78-81 was observed only in the DMSO group on the afternoon of the day of proestrus. At PND 115, randomly selected rats from each of three treatment groups were bilaterally ovariectomized (ovx), and 8 days later, Silastic capsules containing estradiol-17beta were implanted under the skin. Estrogen implants stimulated afternoon surges of LH, FSH, and PRL for two consecutive days in the DMSO group, but not in the OP and estradiol groups. Rats from the OP and DMSO groups underwent ovx at PND 186, and 6 days later they were treated with a combination of estradiol benzoate s.c. (15 microg/kg BW) and progesterone s.c. (2 mg/kg BW) to test the lordosis reflex. In response to this hormone treatment and mounting stimulus delivered by the stud male rats, the OP-treated rats were less receptive compared with control DMSO-treated rats, and thus the lordosis quotient and lordosis rating were significantly (P < 0.05) reduced in the OP group compared with the DMSO group. Analysis of the area of the sexually dimorphic nucleus of the preoptic area of the brain revealed that the area of this nucleus was larger in the OP group than it was in control DMSO rats. We conclude that the exposure of neonatal female rats to higher doses of OP disrupts the cyclic release of LH, FSH, and PRL, and interferes with the display of sexual receptive behavior in adulthood.

Testicular secretion of inhibin in the male golden hamster (Mesocricetus auratus).

To identify the cellular source of inhibin in the male golden hamster, we have used complementary approaches, immunohistochemistry and enzyme-linked immunosorbent assay (ELISA). Strong positive staining of the inhibin alpha subunit was observed in both the Sertoli and Leydig cells of the testes. No specific staining was observed for the inhibin betaA subunit, whereas specific staining for the inhibin betaB subunit was strongly positive in the Leydig cells. Inhibin pro-alphaC and inhibin B were detected in peripheral plasma, and testicular homogenate also contained large amounts of inhibin pro-alphaC and inhibin B. However, inhibin A was not detected either in peripheral plasma or in testicular homogenate. Plasma concentrations of inhibin pro-alphaC and inhibin B were significantly (P < .001) decreased 24 hours after orchidectomy. These results strongly suggest that the Leydig cells are the main source of dimeric inhibin B in the male golden hamster.

The cytotoxic effects of diesel exhaust particles on human pulmonary artery endothelial cells in vitro: role of active oxygen species.

Diesel exhaust particles (DEP) have been proved to induce serious pulmonary injury, among which lethal pulmonary edema has been assumed to be mediated by vascular endothelial cell damage. In the present study, we investigated the cytotoxic mechanism of DEP on human pulmonary artery endothelial cells focusing on the role of active oxygen species. Endothelial cell viability was assessed by WST-8, a novel tetrazolium salt. Nitric oxide (NO) production was measured by using a new fluorescence indicator, diaminofluorescein-2 (DAF-2). Organic compounds in DEP were extracted by dichloromethane and methanol. DEP-extracts damaged endothelial cells under both subconfluent and confluent conditions. The DEP-extract-induced cytotoxicity was markedly reduced by treatment with SOD, catalase, N-(2-mercaptopropionyl)-glycine (MPG), or ebselen (a selenium-containing compound with glutathione peroxidase-like activity). Thus superoxide, hydrogen peroxide, and other oxygen-derived free radicals are likely to be implicated in DEP-extract-induced endothelial cell damage. Moreover, L-NAME and L-NMA, inhibitors of NO synthase, also attenuated DEP-extract-induced cytotoxicity, while sepiapterin, the precursor of tetrahydrobiopterin (BH(4), a NO synthase cofactor) interestingly enhanced DEP-extract-induced cell damage. These findings suggest that NO is also involved in DEP-extract-mediated cytotoxicity, which was confirmed by direct measurement of NO production. These active oxygen species, including peroxynitrite, may explain the mechanism of endothelial cell damage upon DEP exposure during the early stage.

Development of embryos in superovulated guinea pigs following active immunization against the inhibin alpha-subunit.

Embryo recovery and subsequent embryonic development from guinea pigs treated with or without inhibin vaccines were compared to determine the effect of active immunization against the inhibin alpha-subunit. Twenty female guinea pigs of the Hartley strain were injected 3 times either with 1 ml inhibin vaccine (recombinant ovine inhibin a-subunit in oil emulsion: 50 microg/ml, inhibin-immunized group), or 1 ml placebo (saline in oil emulsion; control group) at 4 week intervals. After one estrous cycle following the last injection, females were naturally mated and embryos were collected at 11:00 hr of day 6 of pregnancy (Day 1: sperm in the vaginal smear) for culture in vitro. Active immunization increased the number of corpora lutea (12.6+/-3.0 vs. 4.6+/-0.2, P<0.05), recovered embryos (9.8+/-1.9 vs. 3.6+/-0.4, P<0.01) and normal embryos (7.8+/-1.4 vs. 3.6+/-0.4, P<0.05), although estrous cycle length was not affected (P>0.05). During subsequent 8 day culture in vitro, most of the recovered embryos formed trophoblast outgrowth; 100% (14/14) and 88.2% (15/17) in control and immunized groups, respectively. High levels of inhibin antibody titers were sustained in the inhibin-immunized guinea pigs at least for 5 months after the last injection while no antibody titer was detected in the control animals. These results indicate that active immunization against the inhibin a-subunit is a long-acting and efficient method to induce superovulation with normal embryonic development in the guinea pig.

Follicle selection in cyclic guinea pigs with active immunization against inhibin alpha-subunit.

Experiments were conducted to elucidate the mechanisms of active immunization against inhibin on ovarian follicular development and selection in guinea pigs. Estrous cycle was synchronized in experimental guinea pigs by implanting progesterone containing tubes. Antibodies that bound 125I-labeled bovine inhibin were produced by all guinea pigs receiving the inhibin vaccine (recombinant ovine alpha-subunit in oil emulsion) without any effects on duration of the estrous cycle. Active immunization against inhibin increased the plasma concentrations of progesterone during the luteal phase and the plasma concentrations of estradiol but failed to increase the plasma concentration of follicle-stimulating hormone (FSH) during preovulatory period. The treatment also increased the number of corpora lutea (from 1.3+/-0.3 to 7.0+/-1.6 per each ovary), and preovulatory sized follicles (from 1.8+/-0.6 to 7.0+/-1.6 per each ovary), and follicles stained positively for inhibin alpha-subunit (from 2.3+/-0.5 to 6.3+/-1.3 per each ovary) significantly. The results indicate that active immunization against inhibin enhances ovulation rate by affecting the follicle selection and only dominant follicle can be stained for inhibin alpha-subunit in guinea pigs. This study is firstly to provide direct evidence that inhibins play important role in follicle selections in guinea pigs.

Induction of superovulation by inhibin vaccine in cyclic guinea-pigs.

Experiments were conducted to determine whether neutralizing endogenous inhibin affects follicular development and ovulation rate in guinea-pigs. Eighteen female guinea-pigs bearing 4 week progesterone implants were divided into three groups. At 1 week after removal of the progesterone implants, the animals were given a s.c. injection of 1 ml placebo (saline in oil emulsion; control), or 25 or 50 micrograms inhibin vaccine three times at 4 week intervals. Blood samples were collected once a week throughout the experiment for measuring inhibin antibody titres. After the third injection of inhibin vaccine, blood samples and ovaries were collected on the morning of day 8 after the day of oestrus. Inhibin vaccine increased the ovulation rate in a dose-dependent manner (placebo: 4.2 +/- 0.4; 25 micrograms inhibin vaccine: 6.2 +/- 0.9; 50 micrograms inhibin vaccine: 9.8 +/- 0.9) without any effects on the duration of the oestrous cycle. The results also showed that active immunization against inhibin increased the number of atretic follicles of 300-399 microns in diameter on day 8 after ovulation. The present study is the first to show that the active immunization against inhibin may be a useful method for inducing multiple ovulation in guinea-pigs.

Electrocardiographic changes induced by diesel exhaust particles (DEP) in guinea pigs.

We have previously reported that diesel exhaust particles (DEP) caused a negative inotropic effect that was followed by cardiac arrest in the isolated atrial preparation of guinea pigs. The purpose of this study was to examine the systemic effects of DEP on electrocardiographic (ECG) changes using guinea-pigs. We found that intravenously administered dimethyl sulfoxide (DMSO) extract of DEP solution induced arrhythmias and deaths via complete atrioventricular (AV) block in guinea pigs. The LD of DEP solution was 132.0 +/- 7.2 mg/kg. The coefficient of variance (CV) of LD measured by the modified Hatcher-Magnus method was relatively small (5.5%). Fractions of DEP extracted by hexane, ethanol or methanol, 4-hydroxyphthalic acid 2-methyl ester, a compound isolated from methanol extract of DEP did not induce significant ECG changes in guinea pigs. As compared with fresh DEP solution, the DMSO/DEP solution used in the present study induced similar cardiac toxicity after being stored in a freezer at 4 degrees C for 3 days. These results suggest that stable and water-soluble fractions of DEP may be responsible for cardiotoxicity.

Dermatophilosis (Cutaneous streptothricosis) in Kafue lechwe (Kobus leche kafuensis).

Extensive dermatitis caused by Dermatophilus congolensis was identified in two kafue lechwe (Kobus leche kafuensis) in Lochinvar National Park of Zambia. The lesions were characterized by thickening of the skin, crusts, and nodfule formation. Almost all parts of the body were affected. Histologically there was an exudative dermatitis with acanthosis, parakeratosis, hyperkeratosis, and an exudate rich in neutrophils. This is the first known report of dermatophilosis in lechwe.

[Histopathological changes induced by thermal exposure in the rat liver].

To clarify the effects of a thermal environment on rats, histopathological changes, and temperatures in the rectum and eyes of Fischer strain rats exposed to a thermal environment of 32 degrees C for 2 days or 7 days were investigated. Thermal exposure to 32 degrees C induced elevations in rectal and ocular temperatures. The increased temperature levels persisted during the thermal exposure. Vacuolar degeneration surrounding the hepatic vein was observed in rat livers exposed to 32 degrees C for 7 days. No abnormalities were observed in other organs. These results indicate that thermal exposure to 32 degrees C induces vacuolar degeneration in hepatocytes due to the elevation of body temperature.

Effects of ozone and nitrogen dioxide on drinking and eating behaviors in mice.

Male ICR mice were exposed continuously to ozone (O3) and nitrogen dioxide (NO2) for 7 days to examine the effects on drinking and eating behaviors. Ozone at 0.1 ppm did not affect drinking and eating activities, whereas drinking activity decreased in a concentration-dependent manner to 47.7, 12.8, and 3.0% of the control value with 2-day exposures to 0.2, 0.4, and 0.8 ppm O3, respectively, and eating activity decreased to 35.2 and 8.7% of the control value at 0.4 and 0.8 ppm O3, respectively. Body weight also decreased markedly by 2.0, 4.6, and 7.5 g at 0.2, 0.4, and 0.8 ppm O3, respectively. These decrements reached a maximum on the second day of exposure. However, alterations in drinking and eating activities and body weight were transient, leading to recovery during the continuous O3 exposures. The recovery processes were dependent on the concentrations of O3. Nitrogen dioxide at 4 ppm did not affect drinking and eating activities, whereas drinking activity decreased in a concentration-dependent manner to 56.8, 8.3, and 18.7% of the control value with 2-day exposures to 6, 8, and 12 ppm NO2, respectively, and eating activity decreased markedly to 21.8 and 16.4% at 8 and 12 ppm NO2, respectively. Body weight also decreased by 2.5, 5.5, and 6.1 g at 6, 8, and 12 ppm NO2, respectively. These decrements reached a maximum on the second day of exposure. As in the O3 exposures, the decrements in drinking and eating activities and body weight were transient and recovered during the continuous exposures to NO2 depending on the concentrations of NO2. Drinking and eating activities and body weights of mice that had been previously exposed to 12 ppm NO2 for 7 days did not show changes when the mice were exposed to 0.4 ppm O3 9 days after NO2 exposure. The present study demonstrates that photochemical oxidants suppress drinking and eating behaviors in mice and that they recover thereafter under the continuous exposure conditions.

Effect of ozone toxicity in the drinking behavior of rats.

To clarify the effects of ozone on behavior, drinking activity was observed continuously in animals exposed to ozone at concentrations of 0.2, 0.4, and 0.8 ppm for 1 wk. Drinking decreased considerably on the first day after onset of exposure. On the first day the suppression rate was calculated to be 28.1%, 69.5%, and 93.1%, for the 0.2 ppm, 0.4 ppm, and 0.8 ppm group, respectively. The suppression of drinking and recovery were dependent on the concentration of ozone.

Changes of cardiac and respiratory rhythm in non- and tracheostomized rats exposed to nitrogen dioxide.

Cardiac and respiratory changes in non- and tracheostomized rats were examined during exposure to 20 ppm of NO2 for 150 min. The abnormal respiratory pattern consisted of rapid shallow breathing, deep breathing, and apnea, and the bradyarrhythmias were observed in the tracheostomized rats during exposure. Also, similar changes were seen in the nontracheostomized rats. A decrease in the heart rate (HR) was observed in both non- and tracheostomized rats. The decrease in HR was depressed by atropine injection, and the abnormal respiratory patterns were almost abolished by this drug. It was suggested, from these results, that the cardiac and respiratory abnormalities could be induced without the irritation to upper respiratory tracts, and that the vagal efferent pathway had an important role in the appearance of the abnormalities during exposure.

Reflex cardiopulmonary responses by stimulation to type J receptors in rats exposed to NO2.

To examine the role of the vagal pathway on the cardiopulmonary functions in NO2-exposed rats, phenyl diguanide, which stimulates type J receptors in the lungs, was injected to control and exposed rats at a constant dose. Based on a statistical test, a decrease in the heart rate (HR) after the injection was observed in the groups exposed to 20 ppm NO2 for 1.5 h and 3h, and 10 ppm for 24 h. On the other hand, an increase in respiratory rate (RR) was observed in the groups exposed to 10 ppm for 3 h and 4 ppm for 1 wk. No change in HR and RR was found in the group exposed to 0.4 ppm for 4 wk. These results suggest that the augmentation of the reflex cardiopulmonary responses due to stimulation to the type J receptors was produced by exposures with a higher concentration of NO2.