Exploration for unknown substances in rapeseed oil that shorten survival time of stroke-prone spontaneously hypertensive rats. Effects of super critical gas extraction fractions.

To identify the causative substances for the shortening of survival time by rapeseed (Canola) oil in stroke-prone spontaneously hypertensive rats (SHRSP), SHRSP were fed on a standard chow supplemented with 10 w/w% soybean oil (control), rapeseed oil, one of the fractions of rapeseed oil obtained by super critical gas extraction (SCE) under a pressure of 180-bar or 350-bar, at 40 degrees C, or the residue from the extraction (with 0.5% NaCl in drinking water). In another series of experiment, SHRSP were fed for 8 weeks on the above-mentioned diets without salt loading and autopsied.

Detection of initiating as well as promoting activity of chemicals by a novel cell transformation assay using v-Ha-ras-transfected BALB/c 3T3 cells (Bhas 42 cells).

Cell transformation assay using BALB/c 3T3 cells, C3H10T1/2 cells and others, can simulate the two-stage carcinogenesis utilized for formation of transformed foci. A sensitive cell transformation assay for tumor initiators as well as promoters has been developed using a v-Ha-ras-transfected BALB/c 3T3 cell line, Bhas 42; these cells are regarded as initiated in the two-stage paradigm of carcinogenesis. To distinguish between initiation and promotion, the initiation assay involves a 2-day treatment of low-density cells, obtained one day after plating, with a test chemical, and the promotion assay involves treatment of near-confluent cells with a test chemical for a period of 12 days (Day 3-14).

Influence of di-(2-ethylhexyl)phthalate on fetal testicular development by oral administration to pregnant rats.

Influence of di-(2-ethylhexyl)phthalate (DEHP) on testicular development was studied by oral administration of DEHP at doses of 500 and 1000 mg/kg/day to pregnant rats on gestational days (G) 7 to 18. Ethinyl estradiol (EE) at dose levels of 0.25 and 0.

Acute functional enhancement of circulatory neutrophils after intratracheal instillation with diesel exhaust particles in rats.

We have previously demonstrated that intratracheal instillation (IT) with diesel exhaust particles (DEP) exacerbates myocardial ischemia/reperfusion-induced arrhythmia in rats. Since activated neutrophils play a pivotal role in ischemia/reperfusion arrhythmia, in the present study we investigated the effects of DEP on peripheral neutrophil count and on the oxyradical production (ORP) of neutrophils in rats. We also determined the production of cytokines for better understanding of the relationship between pulmonary inflammation and neutrophil function.

Delayed exacerbation of acute myocardial ischemia/reperfusion-induced arrhythmia by tracheal instillation of diesel exhaust particles.

For understanding the relationship between the increased incidence of sudden cardiac death and air pollution, we examined the effects of intratracheal instillation of diesel exhaust particles (DEP) on acute myocardial ischemia/reperfusion-induced arrhythmia in rats. The animals received 1 mg DEP 24-48 h before the ischemia/reperfusion (DEP-pretreated group, DEP-PRE), and were subjected to 3 successive brief ischemia/reperfusion (3 min ischemia followed by 5 min reperfusion) procedures. These were to make the animals tolerant to ischemia/reperfusion-related myocardial deterioration.

Locomotor hyperactivity following prenatal exposure to 5-bromo-2'-deoxyuridine: neurochemical and behavioral evidence of dopaminergic and serotonergic alterations.

Prenatal exposure to 5-bromo-2'-deoxyuridine (BrdU) has been reported to induce abnormal behaviors in offspring, including marked hyperactivity. In this study, the contribution of the serotonin (5-HT) and dopamine (DA) systems to BrdU-induced developmental neurotoxicity was investigated. Sprague-Dawley rats were treated with BrdU on gestational days 9 through 15 (50mg/kg, i.

Subcellular distribution of di-(2-ethylhexyl)phthalate in rat testis.

Subcellular distribution of di-(2-ethylhexyl)phthalate (DEHP) in the testis was studied by single oral administration of [3,4,5,6-(3)H]-phthalic acid di-(2-ethylhexyl) ester (DEHP-3H) or phthalic acid di-(2-ethyl[1-(3)H]hexyl) ester (3H-DEHP) to 8-week-old male rats. Autoradiographs and electron microscopic autoradiographs were prepared from the testis, liver and kidney at 6 and 24 hr after administration and distribution of radioactive materials in the tissues were observed. In the autoradiographic specimen at 6 hr after administration of DEHP 3H-labeled at phthalic acid moiety (DEHP-3H), many grains were observed in the testis, mainly at the basal area of seminiferous tubules at the stages IX to I of the spermatogenic cycle.

Cell transformation assay using Balb/c 3T3 cells or Bhas 42 cells for the efficient detection of tumour promoters.

In this paper, we report three interlaboratory validation studies with special interest in detecting tumour promoters. The former two validation studies were on the improved two-stage in vitro transformation assay using Balb/c 3T3 cells. The third one was on the Bhas 42 cell assay.

Validation activities in Japan: a report.

Japanese activities in the validation of alternative test methods are reviewed. The validation of the Limulus endotoxin test as an alternative method to the rabbit pyrogen test was initiated by the Japanese Pharmacopoeia Society, and was successfully conducted under the direction of the National Institute of Health Sciences (NIHS), with the participation of many pharmaceutical company laboratories. The validation of in vitro alternatives to the Draize eye test for cosmetics were carried out by the NIHS and cosmetic company laboratories.

Enzyme-linked immunosorbent assay for monitoring toxic dioxin congeners in milk based on a newly generated monoclonal anti-dioxin antibody.

To develop an enzyme-linked immunosorbent assay (ELISA) for monitoring the toxicity due to polychlorinated dibenzo-p-dioxins and dibenzofurans contaminated in human breast milk, we have generated novel monoclonal antibodies using some haptenic derivatives linked to bovine serum albumin via the C-1 or C-2 position on the dioxin skeleton. BALB/c or A/J mice were repeatedly immunized with the immunogen, and spleen cells were fused with P3/NS1/1-Ag4-1 myeloma cells. After five fusion experiments, a hybridoma clone was established that secretes an antibody D9-36 group specifically recognizing the major toxic congeners, 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD), 1,2,3,7,8-pentachlorodibenzo-p-dioxin, and 2,3,4,7,8-pentachlorodibenzofran.

Minor involvement of somatic growth in the onset of puberty of Hatano high- and low-avoidance rats.

The present study was planned to examine the effects of somatic growth on the determination of timing of puberty using Hatano high- and low-avoidance rats (HAAs and LAAs); the rats were genetically selected from Sprague-Dawley (SD) rats for good or poor performance in a two-way active avoidance-learning test. Since these two lines were found to have different characteristics, such as body weight at birth, maternal care and timing of male puberty, the present study characterized female puberty in Hatano rats and then compared postnatal growth and timing of puberty between the two lines of rats when they were nursed by foster SD dams. When nursed under biological dams, HAAs became heavier, exhibited vaginal opening at a younger age and first ovulation was accompanied by more oocytes than LAAs.

An improved technique for repeated gavage administration to rat neonates.

The technique for gavage administration to rat nurslings was improved to allow determination of the direct effects of chemical substances in the nurslings. Rat neonates were treated with distilled water from postnatal day 1 through 20 using this technique. The viability of neonates during the administration period was comparable to that of untreated neonates.

Remnant-like particles from subjects who died of coronary artery disease suppress NO synthase activity and attenuate endothelium-dependent vasorelaxation.

Background: Plasma levels of remnant-like particles (RLP) is one of the predictive markers for coronary artery disease (CAD), and the inhibition by RLP of endothelium-dependent vasorelaxation has been reported. We attempted to clarify whether or not RLP, which inhibits endothelium-dependent vasorelaxation, affects nitric oxide (NO) production and NO synthase (eNOS) levels in cultured endothelial cells.

Methods: RLP were obtained from postmortem blood of subjects who died of CAD.

Rapeseed oil ingestion and exacerbation of hypertension-related conditions in stroke prone spontaneously hypertensive rats.

Two groups of 20 stroke prone spontaneously hypertensive rats (SHRSP) at 5 weeks old were fed a diet containing 10 w/w% rapeseed (canola) oil or soybean oil as the only dietary fat, and given drinking water containing 1% NaCl. Life span of the canola oil group (62+/-2 days) was shorter than that of the soybean oil group (68+/-3 days). Stroke-related symptoms were observed in every animal, but the onset of those in the canola oil group, at 47+/-1 days after starting the administration was earlier than that in the soybean oil group, 52+/-2 days.

Effects of reduction of the number of primordial follicles on follicular development to achieve puberty in female rats.

Effects of reduction of the number of primordial follicles on follicular development and concentrations of circulating hormones were examined in immature female rat offspring of dams given busulfan intraperitoneally on day 14 of gestation. The offspring of dams treated with 5 mg busulfan kg(-1) showed vaginal opening at an age comparable with the offspring of dams treated with 2.5 mg busulfan kg(-1) or with corn oil as a control, although they exhibited an irregular oestrous cycle until week 14 after birth.

Maternal behavior, milk ejection, and plasma hormones in Hatano high- and low-avoidance rats.

Hatano high- and low-avoidance (in a two-way active avoidance task) animals (HAA and LAA, respectively) were successfully selected from a Sprague-Dawley rat population. Pup growth of both strains was dependent on the maternal strain in a cross-fostering study. To determine whether there are strain-specific differences between HAA and LAA in maternal care, both strains of dams were subjected to a test battery as measured by nest building, home cage behavior, pup retrieval, and milk ejection tests.

Relevance of chemical structure and cytotoxicity to the induction of chromosome aberrations based on the testing results of 98 high production volume industrial chemicals.

Over a 6-year period (1991-1996), the chromosomal aberration testing of high production volume (HPV) industrial chemicals had been conducted using Chinese hamster lung (CHL/IU) cells according to OECD HPV testing program and the national program in Japan. A total of 98 chemicals were tested for the induction of chromosome aberration (CA), consisting of structural CA and polyploidy. Of the 98 chemicals, structural CA and/or polyploidy were induced by 39 chemicals (40%).

Strain differences in the spontaneous incidence of sperm morphological abnormalities in Hatano rats.

Two inbred strains of Hatano rats, which had been bred selectively on the basis for shuttlebox avoidance responses (high- and low-avoidance animals; HAA and LAA), were different in sperm motility. We have found that there are strain differences in the spontaneous incidence of sperm morphological abnormalities. The percentages of abnormalities were extraordinarily higher in the LAA rats, whose sperm motility was lesser.

Low-dose bisphenol A does not affect reproductive organs in estrogen-sensitive C57BL/6N mice exposed at the sexually mature, juvenile, or embryonic stage.

Bisphenol A (BPA) is used on a large scale in the manufacture of polycarbonate plastics. BPA has been shown to bind weakly to both estrogen receptor (ER) alpha and ER beta. The objective of this study was to evaluate the effects of low-dose BPA on male sexual development after exposure at various stages of development.

Differences in sperm motion between high- and low-shuttlebox avoidance rats (Hatano strains).

Sperm from the caudal epididymis of 2 inbred strains of Sprague-Dawley (SD) rats, selected on the basis of their high- or low-shuttlebox avoidance responses, were analyzed for motion characteristics by a computer-assisted sperm motion analysis (CASA) system. Sperm motion in high-avoidance animals (HAA) was characterized by high velocities, high amplitude of lateral head displacement (ALH), and low beat cross frequency (BCF). Conversely, sperm from low-avoidance animals (LAA) displayed low velocities, low ALH, and high BCF.

Effects of endocrine disrupting chemicals on the microtubule network in Chinese hamster V79 cells in culture and in Sertoli cells in rats.

The effects of ten endocrine disrupting chemicals, i.e., bisphenol A (BPA), p-nonylphenol (NP), p-octylphenol (OP), p-pentylphenol (PP), butyl benzyl phthalate (BBP), dicyclohexyl phthalate (DCHP), di-n-butyl phthalate (DBP), tetrabutyltin (TBT), tri-n-butyltin chloride (TBC), and di-n-butyltin dichloride (DBD), as well as 17 beta-estradiol (E(2)) as a positive control on the microtubule network in Chinese hamster V79 cells in culture were examined by the indirect immunofluorescence method using anti-beta-tubulin antibody.

Reproductive effects in male and female rats from neonatal exposure to p-octylphenol.

A number of alkylphenolic compounds are used in a variety of commercial products and have been shown in in vitro studies to be weakly estrogenic, but in vivo data are not available addressing this issue in mammals. Human exposure to alkylphenols may occur not only from these environmental contaminants but also through contact with manufactured and metabolic breakdown products. In this study, Sprague-Dawley rats were exposed to octylphenol by oral gavage at doses of 0 (vehicle: corn oil), 12.

High-performance immunoaffinity chromatography, an immunoaffinity membrane for selective removal of plasma components, and safety evaluation of the latter system.

This article reviews our studies on evaluating the suitability of high-performance immunoaffinity chromatography (HPIAC), an immunoaffinity membrane (IAM) for removing unwanted plasma component, and the safety of the IAM. In an attempt to resolve the problem of amyloid deposition in dialysis patients, we used an HPIAC column, bearing anti-beta 2-microglobulin to remove specifically beta 2-MG from human plasma. The use of a membrane as an affinity ligand support was also studied.

Reproductive effects in male and female rats of neonatal exposure to genistein.

Sprague-Dawley rats were administered genistein orally at doses of 12.5, 25, 50, or 100 mg/kg on postnatal days 1 through 5 to examine its effects on reproductive function after puberty. In addition, preputial separation and vaginal opening as endpoints of sexual maturation, estrous cycling, sperm count, serum testosterone concentration, and histopathologic changes of reproductive organs of male and female rats were examined.