Oral Contraceptives and Increased Formation of Soluble Fibrin

Soluble fibrin was measured weekly for two months in 12 normal women and in 12 women on combined oestrogen-progestogen therapy (Ortho-Novin, Norinyl, Enavid, Ovral, Ovulen, Demulen). Plasma soluble fibrin concentration in women on oral contraceptives showed an increase of 97·2% (P <0·001) above that of normal women. In three cases, where each woman on the oral contraceptive served as her own control, stopping medication led to a return to normal of the plasma content of soluble fibrin.     

Effective suppression of ovarian cyclicity in the lowland gorilla with an oral contraceptive

To determine the effects of a human oral contraceptive on normal and abnormal reproductive endocrine patterns in two lowland gorillas (Gorilla gorilla), daily urine samples were analyzed by radioimmunoassay for estrone conjugates (EC) and pregnanediol-3-glucuronide (PdG). During a pre-treatment period one female (F1) demonstrated regular menstrual cycles averaging 27.6 ± 1.8 days, whereas ovarian cyclicity in female 2 (F2) was consistently prolonged, ranging from 37–51 days. A 56 day regimen of an oral contraceptive (Demulen 50®) was administered to both females beginning in the late luteal phase, and within 6 and 7 days of treatment onset (F1 and F2, respectively) urinary EC and PdG declined to and remained at concentrations consistent with amenorrhea throughout the administration period. Ten and twenty days after contraceptive withdrawal (F1 and F2, respectively) an EC peak was observed with subsequent PdG elevations 1–2 days later. Mean cycle length and luteal phase durations were not different (P > 0.05) before or after treatment for either female, while combined peak PdG concentrations were greater (P < 0.05) for the first 3 months after treatment compared to pre-treatment values. These results indicate that a human oral contraceptive rapidly suppresses ovarian activity in female lowland gorillas, but that the abnormal endocrine pattern observed in F2 could not be redirected into a normal profile after contraceptive withdrawal. © 1992 Wiley-Liss Inc.     

Regional Heterogeneity of Nicotine Effects on Neurotransmitters in Rat Brains <Emphasis Type="Italic">in vivo</Emphasis> at Low Doses

In our recent studies on nicotine-induced changes in neurotransmitters in brain areas associated with cognitive function using a nicotine dose of 0.5 mg/kg administered subcutaneously to conscious freely moving rats, we found changes in dopamine, norepinephrine, and serotonin, and their metabolites, in the areas examined. For the present report we examined changes in these neurotransmitters following administration of lower nicotine doses, to test regional differences in nicotine response and possible threshold levels for some effects of nicotine. The doses used were 0.15 mg/kg and 0.03 mg/kg nicotine administered subcutaneously. Nicotine levels in the brain reached peak values in less than 10 min and decreased with a half-life of about 60 min (0.15 mg/kg) or 30 min (0.03 mg/kg) to values below detection limits (1 ng/g), by the later time points of the 0.03 mg/kg experiments. Nicotine-induced dopamine (DA) increase (and increase in DA metabolites) and decrease in 5-HT levels at 0.15 mg/kg were significant in the cortex, less so in the hippocampus. Norepinephrine (NE) increase at 0.15 mg/kg was much less significant than found previously at 0.5 mg/kg. At a low nicotine dose (0.03 mg/kg), the significant changes observed were a decrease in 5-HT in the hippocampus and small increases of DA and NE in the prefrontal cortex and of NE in the medial temporal cortex. In the nucleus accumbens DA, NE, and 5-HT and their metabolites in the ventral tegmental area, mostly DA and metabolites were increased. We conclude that in areas of cognitive function nicotine-induced DA changes are more concentration dependent than changes in NE or 5-HT, and that there are regional differences in neurotransmitter changes induced by nicotine, with NE changes detectable only in the cortex and 5-HT changes only in the hippocampus at a low nicotine dose, indicating significant regional variation in sensitivity to nicotine-induced neurotransmitter changes in brain areas associated with cognitive function. The decrease in 5-HT shows that nicotine also has indirect effects caused by neurotransmitters released by nicotine. The effects of low nicotine dose are more significant in areas of reward function, indicating differences in sensitivity between cognitive and reward functions.     

Metabolomic profiling of biomarkers of liver X receptor-induced toxicity in mouse liver tissue

Methods based on matrix-assisted laser desorption/ionisation mass spectrometry (MALDI-MS), liquid chromatography coupled to an LTQ-Orbitrap mass spectrometer (LC-MS) and gas chromatography-mass spectrometry (GC-MS) were used to investigate changes in the small molecule profiles of mouse liver in response to administration of an LXR agonist. Mice were treated with either 0.3 mg/kg, 1 mg/kg, 10 mg/kg, 30 mg/kg or 60 mg/kg of an LXR test compound or saline (control) once daily, over a 5 day period, to investigate the effects of the drug on metabolism in the liver. It was possible to detect triacylglycerol accumulation in the livers of animals treated with the drug, even at the lowest concentrations using, in the first instance, MALDI MS. There was also an increase in the relative degree of triacylglycerol saturation in the drug-treated samples. Changes in the profiles of phosphatidylcholine lipids were also observed. The changes in lipid profiles were also confirmed by LC-MS and GC-MS, the latter revealing a large increase in the level of the free fatty acid oleic acid (C18:1) in the treated samples. All of the changes were dose-related. Polar metabolites in the samples were analysed by hydrophilic interaction (HILIC) chromatography in combination with an LTQ-Orbitrap mass spectrometer. There were many changes in the metabolite profiles, some of which might simply be related to generalised toxicity. The clearest marker compounds, which showed very marked changes with dose, were methylglutaryl carnitine (MGC) and hydroxymethylglutaryl carnitine (HMGC). Another marker of some interest was uridine diphosphate N-acetylglucosamine (UNGA).     

Subclinic effect of the administration of T-2 Toxin and Nivalenol in mice

Four experiments using T-2 toxin and nivalenol at different dosage, which represented the 25% and 40% of the LD50 (experiment A: 1.04 mg of T-2 toxin per kilogram of body weight, experiment B: 2.34 mg of T-2 toxin/kg b.w., experiment C: 1.04 mg of T-2 toxin/kg b. w. and 2.34 mg of T-2 toxin/kg b.w.; experiment D: 0.82 mg of nivalenol/kg b.w. and 1.845 mg of nivalenol/kg b.w.) were conducted on 400 mice. Both toxins were administered to mice of different ages (experiments A and B were adults, experiment C and D were young) by intraperitoneal single injection, and the clinical signs, hematological variables and histoanatomo pathological changes were studied. All animals survived. No changes anatomo-histopathological nor significative differences in weight gain were observed. Different behaviors were found for nivalenol and T-2 toxin. The most significant change was the increase in the level of monocytes in old animals, so this could be a biological indicator for T-2 toxin subclinical intoxication.     

A 1H NMR spectroscopic study of the biochemical effects of ifosfamide in the rat: evaluation of potential biomarkers

Ifosfamide (55 mg kg^-1 and 110 mg kg^-1) was administered via single i.p. injections to Sprague-Dawley rats and urine samples were collected for the periods of -24-0, 0-8 h, 8-24 h, 24-48 h and 48-72 h post-dose. Quantitative changes in the excretion pattern of small organic molecules in the urine of rats treated with ifosfamide were studied using high frequency 1H NMR spectroscopy. The kidneys and livers of the animals were also examined, but showed no marked histopathological changes. ¹     

Proteomic analyses of lung lysates from short-term exposure of Fischer 344 rats to cigarette smoke

A short-term 5 day mainstream cigarette smoke exposure study was conducted in Fischer 344 rats to identify changes in lung proteins. Groups of 10 male and female rats at 5 weeks of age were assigned to one of four exposure groups. Animals received either nose-only filtered air (Air Control) or 75, 200, or 400 mg total particulate matter (TPM)/m(3) of diluted cigarette smoke. Exposures were conducted for 3 h per day, for 5 consecutive days. One lung per animal was frozen in liquid nitrogen and processed for proteomic analyses. Lung lysates from control verses treated animals were screened with 650 antibodies for changes in signaling protein levels and phosphorylation using antibody microarray technology, and then over 100 of the top protein hits were assessed by immunoblotting. The top smoke-altered proteins were further evaluated using reverse lysate microarrays. Major protein changes showed medium to strong bands on Western blots, depended on dose and gender, and included protein-serine kinases (Cot/Tpl2, ERK1/2, GSK3α/β, MEK6, PKCα/γ, RSK1), protein phosphatases (PP4/A'2, PP1Cβ), and other proteins (caspase 5, CRMP2, Hsc70, Hsp60, Rac1 and STAT2). The most pronounced changes occurred with 75 mg TPM/m(3) exposed females and 200 mg TPM/m(3) exposed males. Smoke-altered proteins regulate apoptosis, stress response, cell structure, and inflammation. Changes in identified proteins may serve as early indicators of lung damage.     

An application of Lacker's mathematical model for the prediction of ovarian response to superstimulation

Introduction. A mathematical model of ovarian follicular growth is applied to the problem of predicting ovarian response in a superstimulation protocol. Methods. Fifty-four women enrolled in an ovarian superstimulation program of therapy for the amelioration of idiopathic infertility had their ovarian cycles synchronized by taking Demulen 30 for two weeks prior to the study. Daily ultrasonographic imaging, measurements of serum estradiol and doses of hMG began on day 5 after the patients stopped taking Demulen. The diameters of individual follicles were measured and followed daily. When the largest follicle attained a diameter of 19 mm, hCG was given to induce ovulation. Individual follicle growth data were fit to a mathematical model of ovarian follicle maturation and the resulting parameters were used to classify patients into low and high ovarian response groups. Results. The parameters computed from the mathematical model fit were found to be predictive of ovarian response with a sensitivity of 71% and a specificity of 70%. The parameters were also meaningful within the context of the original mathematical model and have value for determining how doses of hMG may be adjusted during the course therapy to increase the ovarian response in individuals. Conclusion. Mathematical modeling of ultrasonographically derived follicular growth data has significant potential for clinical application in ovarian superstimulation protocols. The method of fitting follicular growth data to a mathematical follicle maturation surface furthermore provides a straightforward approach for the characterization of ovarian follicular dynamics in general.     

Cholinergic Deafferentation of Dorsal Hippocampus by Fimbria-Fornix Lesioning Differentially Regulates Subtypes (m1-m5) of Muscarinic Receptors

Abstract: Unilateral aspiration lesions of the rostral supracallosal stria/cingulum bundle and fimbria-fornix were performed on adult female rats. Ten and 24 days post lesioning, an elevation (17%; p<0.01) of total muscarinic receptors was observed in lesioned versus control hippocampi. By using antisera selective for each of the five molecularly defined subtypes (m1-m5) of muscarinic receptors, significant changes were observed in the levels of expression for at least four receptor proteins. Three receptor subtypes increased in density: m1 by 14% (from 943 to 1,078 fmol/mg); m3 by 77% (from 150 to 268 fmol/ mg); and m4 by 29% (from 220 to 285 fmol/mg). In contrast, a 22% decrease in the density of m2 receptors was found (from 220 to 173 fmol/mg). Detectable levels of m5 receptors were low in the hippocampus (∼1% of total receptors), and reliable measurements were not obtained. The directions of these changes are likely to be related to the pre- or postsynaptic localization of these receptor subtypes.     

A 1H NMR spectroscopic study of the biochemical effects of ifosfamide in the rat: evaluation of potential biomarkers

Ifosfamide (55 mg kg^-1 and 110 mg kg^-1) was administered via single i.p. injections to Sprague-Dawley rats and urine samples were collected for the periods of -24-0, 0-8 h, 8-24 h, 24-48 h and 48-72 h post-dose. Quantitative changes in the excretion pattern of small organic molecules in the urine of rats treated with ifosfamide were studied using high frequency 1H NMR spectroscopy. The kidneys and livers of the animals were also examined, but showed no marked histopathological changes. ¹     

Body composition and physiological casein and wheat gluten protein requirements of 180-day-old rats

Six-month-old male rats were given diets with mounting casein and wheat gluten protein concentrations from 0% to 40% and after 14 days the optimum and minimum physiological doses were determined from changes in body nitrogen, body water, body weight and protein intake. The optimum dose for casein protein was 1.54 g/d (5% protein in the diet) and for wheat gluten protein 2.10 g/d (7.5% protein in the diet). The minimum casein and wheat gluten protein doses for 180- to 194-day-old rats, determined from body nitrogen changes, were 1499 mg/d (4.86%) and 1995 mg/d (7.12%) respectively, from body water changes 1561 mg/d (5.06%) and 2027 mg/d (7.23%) and from body weight changes 1333 mg/d (4.32%) and 1951 mg/d (6.06%). It should be noted that, unlike younger animals aged 35-49, 75-89 and 120-134 days, the optimum and minimum doses for six-month-old rats were approximately the same.     

赤霉素诱导甘蔗节间伸长的效应与相关酶活性的关系

以甘蔗品种'新台糖22号'为试验材料,在伸长初期以200 mg/L GA3进行叶面喷施处理,对照喷清水,研究GA3处理后甘蔗节间糖苷酶、过氧化物酶、过氧化氢酶的变化,以揭示赤霉素诱导甘蔗节间伸长与相关酶活性的关系.结果表明:(1)GA3处理的株高在各个时期显著高于对照,而且在处理后7、14和28 d分别比对照提高了17.32%、14.50%和8.35%,GA3处理引起甘蔗植株表现的高度优势一直保持到后期,节间伸长效果主要是在茎的中部(5～10节).(2)GA3处理后α-葡萄糖苷酶和α-甘露糖苷酶的活性较对照显著下降;POD和β-半乳糖苷酶的活性也略有下降;α-半乳糖苷酶、β-N-乙酰氨基已糖苷酶、过氧化氢酶的活性显著提高;β-葡萄糖苷酶的活性也有一定程度提高.由此推测,外源GA3主要通过调节α-葡萄糖苷酶活性、α-甘露糖苷酶、α-半乳糖苷酶、β-N-乙酰氨基已糖苷酶活性和过氧化氢酶,其次是POD、β-半乳糖苷酶和β-葡萄糖苷酶活性,最终达到节间伸长效果.     

The effects of haloperidol and amphetamine on ascorbic acid and uric acid in caudate and nucleus accumbens of rats as measured by voltammetry in vivo

The ability of haloperidol (0.1 mg/kg) to reduce the amphetamine-induced (2 and 5 mg/kg) increase in ascorbic and uric acid in anterior caudate and in nucleus accumbens was tested using voltammetry in vivo. In both areas, haloperidol reduced the amphetamine-induced increase in uric acid. In both areas, haloperidol only marginally affected the amphetamine-induced increase in ascorbic acid. Amphetamine-induced increases in uric acid were more nearly dose-related than changes in ascorbic acid. Of the two compounds, uric acid seems more likely to be associated with dopamine.     

Influence of short term dietary cholesterol and fat on human plasma Lp[a] and LDL levels.

The relationship between plasma levels of Lp[a] and LDL was examined using dietary regimens. In 81 normolipidemic male outpatients, dietary cholesterol was increased by consuming six eggs per day from a mean (SD) level of 311 (162) to 1430 (198) mg per day. Mean (SD) LDL-cholesterol levels increased from 102 (26) mg/dl to 120 (33) mg/dl (P less than 0.001), while mean (SD) Lp[a] levels were 5.5 (6.1) mg/dl on the basal diet and 5.6 (6.4) mg/dl on the cholesterol-rich diet. No significant correlation was observed between increases in either LDL-cholesterol or apolipoprotein B to Lp[a], nor was there any relationship between individual baseline levels of Lp[a] and dietary-induced changes of Lp[a]. Fourteen of the 81 participants were reexamined under strict nutritional control. Four diets with 40% of calories as fat, but differing in the type of fat and the amount of cholesterol, were administered sequentially to all subjects. As expected, mean (SD) LDL-cholesterol and apolipoprotein B levels were highest on the saturated fat, high cholesterol diet (112 (32) mg/dl and 79 (22) mg/dl) and lowest on the polyunsaturated fat, low cholesterol diet (77 (27) mg/dl and 53 (18) mg/dl). In contrast, mean Lp[a] levels did not significantly change among the four diets (range 4.2-4.9 mg/dl). No correlation of Lp[a] responses with changes in plasma lipids, apolipoproteins, or lipoproteins was observed on any diet. These data suggest that determinants of plasma Lp[a] levels are distinctly different from the determinants of plasma LDL levels in normolipidemic males.     

Results of the negative control chemical allyl alcohol in the 15-day intact adult male rat screening assay for endocrine activity

Development, standardization, and validation of methods to assess the potential of chemicals to disrupt hormonal homeostasis have been the focus of considerable research efforts over the past 10 years. As part of our validation effort, we evaluated the specificity of the 15-day intact adult male rat assay, using a negative control chemical, allyl alcohol, a known hepatotoxicant that was not expected to induce endocrine effects. Male rats were dosed for 15 days via oral gavage with 0, 10, 30, 40, or 50 mg/kg/day allyl alcohol. The endpoints evaluated included final body and organ weights, serum hormone concentrations, and a limited histopathology assessment. No mortality or adverse clinical signs were observed. Mean final body weight for rats in the 50-mg/kg/day dose group was decreased to 90% of control. Mean relative liver weights were increased at 40 and 50 mg/kg/day (115% and 117% of control, respectively). Serum testosterone and DHT concentrations were statistically significantly decreased at 50 mg/kg/day (72% of control). Serum prolactin concentrations were statistically significantly decreased at 40 mg/kg/day (58% of control), but not at 50 mg/kg/day. There were no effects on the other endpoints evaluated. Consistent with previous guidance for interpreting the 15-day intact adult male rat assay, histological and weight changes of target organs were given a higher weight-of-evidence than changes in serum hormone concentrations alone. Therefore, with only minimal changes in serum hormone concentrations and no effects on organ weights or microscopic alterations, the results of allyl alcohol in the 15-day intact adult male rat assay were considered negative and consistent with the predicted results.     

Long-term gentamicin ototoxicity in the rat: research on retro-cochlear involvement by study of evoked auditory potentials of the brain stem

Four groups of rats treated with single (50 mg/kg and 200 mg/kg) or multiple (3 X 200 mg/kg and 5 X 200 mg/kg) doses of gentamycin were studied over a 6-month period and compared with a control group. At 50 mg/kg no significant changes were observed. Significant signs of intoxication were observed at 200 mg/kg doses and at multiple doses. A significant lengthening of the latencies with changes in amplitude and a transitional increase in the auditory threshold were observed as early as the end of treatment. This was followed by a phase of amplitude decrease, a rise in auditory threshold and a decrease in latencies which remains unexplained.     

Gonadotropin-releasing hormone binding sites in ovaries of normal cycling and persistent-estrus rats

The gonadotropin-releasing hormone (GnRH) binding capacity in ovaries and pituitaries of normal cycling rats at different stages of the estrous cycle and in ovaries of persistent-estrus rats was measured using radioligand-receptor assay (RRA). Persistent estrus was induced either by neonatal administration of testosterone propionate (1.25 mg s.c.) on the second day of life or by a hypothalamic suprachiasmatic frontal cut made with Halász' knife. All animals were killed during the critical period (1400-1600 h), and GnRH receptor was assayed. GnRH receptor levels in both ovaries and pituitaries changed during the estrous cycle. The total number of ovarian GnRH binding sites was significantly higher in proestrus than in diestrus 1, the stage in which the lowest level was found. When binding sites were expressed in fmol/mg ovary, the highest level was observed in diestrus 2; however, no changes were observed during the estrous cycle when GnRH binding sites were expressed as fmol/mg protein. Changes noted were very similar to those demonstrated in pituitary GnRH receptors in our present and previous experiments. Higher levels of pituitary binding sites were found in diestrus 2 and proestrus than in estrus and diestrus 1. The changes in the GnRH receptor levels were more striking in the pituitary than in the ovaries. It appears that the total number of ovarian GnRH binding sites was not altered in either of the two persistent-estrus groups, but that their concentration was significantly higher (expressed in fmol/mg ovary or fmol/mg protein) than on any day during the estrous cycle.(ABSTRACT TRUNCATED AT 250 WORDS)     

Changes in enzyme activity of corn seedlings after foliar application of triacontanol

Changes in the activity of several enzymes in corn seedlings (Zea mays L.) after 1-triacontanol (TRIA) application have been analyzed. The specific activity of isocitrate dehydrogenase and 6-phosphogluconate dehydrogenase in corn seedlings treated with TRIA increased rapidly. Three days after treatment, the TRIA-treated seedlings showed 89% and 39% more ICDH and 6PGDH activity per mg protein, respectively, than the untreated plants. Malate dehydrogenase activity increased in treated plants at a rate approximately equivalent to the increase in soluble protein. Acid phosphatase, peroxidase, and alkaline phosphatase activity remained relatively constant on a per plant basis and decreased slightly on a per mg protein basis. No qualitative changes were observed in the isozyme patterns of the enzymes analyzed by starch gel electrophoresis, although quantitative changes consistent with the increases using spectrophotometric assays were observed.     

Production of indole diterpenes by Aspergillus alliaceus

Production of two related indole diterpenes (differing by a dimethyl leucine side chain) by Aspergillus alliaceus was improved through several pilot scale fermentations. Media were optimized through focus primarily on initial increases, as well as mid-cycle additions, of carbon and nitrogen sources. Fermentation conditions were improved by varying ventilation conditions using various combinations of air flowrate and back-pressure set points. Production improvements were quantified based on total indole diterpene concentration as well as the ratio of the major-to-minor by-product components. Those changes with a positive substantial impact primarily on total indole diterpene concentration included early cycle glycerol shots and enhanced ventilation conditions (high air flowrate, low back-pressure). Those changes with a significant impact primarily on ratio included higher initial cerelose, soybean oil, monosodium glutamate, tryptophan, or ammonium sulfate concentrations, higher broth pH, and enhanced ventilation conditions. A few changes (higher initial glycerol and monosodium glutamate concentrations) resulted in less notable and desirable titer or ratio changes when implemented individually, but they were adopted to more fully realize the impact of other improvements or to simplify processing. Overall, total indole diterpene titers were improved at the 600 L pilot scale from 125-175 mg/L with a ratio of about 2.1 to 200-260 mg/L with a ratio of about 3.3-4.5. Thus, the ability to optimize total indole diterpene titer and/or ratio readily exists for secondary metabolite production using Aspergillus cultures.     

Dose of FSH-P as a source of variation in embryo production from superovulated cows

Various superovulation treatments were evaluated retrospectively in a commercial embryo transfer program. When it appeared that embryo production was dependent on the dose of FSH-P, a dose response curve to FSH-P was developed and embryo production compared using several treatment regimes. There was a significant effect of dose of FSH-P on embryo production in superovulated cows. At doses in excess of 28 mg, embryo production declined from 5.9 transferable embryos per collection (28 mg) to 2.7 (60 mg). Total embryos collected declined from 14.9 to 6.8 and the percent transferable from 57% to 40%. There was no advantage in using a five-day treatment over a four-day treatment regimen or in using a level over a declining dose regimen. There was a large individual variation in cow response rendering decisions on treatment changes based on single records unreliable. The percentage of zero collections increased with dose rate. Adoption of a 28-mg dose rate in commercial donors resulted in the embryo production forecast by these studies.