The influence of self-focused attention on heartbeat perception.
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Adult↗
Biomedical subjects
J Weisz
Publications and source records attributed to J Weisz.
Identification of microsomal, organic hydroperoxide-dependent catechol estrogen formation: comparison with NADPH-dependent mechanism.
Exogenous aromatic polycyclic hydrocarbons, notably benzo[a]pyrene, can be metabolized by both NADPH-dependent monooxygenase(s) and an organic hydroperoxide (OHP)-dependent mechanism. To determine whether phenolic estrogens, endogenous aromatic polycyclic hydrocarbons, can also be hydroxylated by these alternative pathways, conversion of estradiol (E2) to 2- and 4-hydroxylated catecholestrogens (2- and 4-OH-CEs) by human placental microsomes was examined under conditions previously shown to support CE formation by NADPH- and OHP-dependent mechanisms. CEs were formed under both conditions. Properties of OHP-dependent activity were similar to CE formation by peroxidases and distinct from the NADPH-dependent monooxygenase. NADPH supported only 2-hydroxylation, whereas cumene hydroperoxide supported 2- and 4-hydroxylation equally. Monooxygenase-mediated activity had characteristics of a high-affinity, low-capacity enzyme system (apparent Km for E2 = 0.3 microM, and Vmax = 31 pmol/mg protein/30 min), whereas peroxidatic activity had properties of low affinity and high capacity (apparent Km for E2 = 55 microM, Vmax = 666 pmol/mg protein/10 min). The requirement of peroxidatic activity for oxidizing co-substrate could be met by OHPs but not by H2O2. Peroxidatic CE formation could have special functional significance for physiological and pathological consequences of estrogen action since it generates 4-OH-CEs which are both effective catechols and potent, long-acting estrogens. Moreover, it could provide a link through co-oxygenation between estrogens and diverse cellular mechanisms involving generation of OHPs.
Carbon Dioxide↗
Characteristics of estrogen-2/4-hydroxylase of porcine ovarian follicles: influence of steroidal and non-steroidal agents on the activity of the enzyme in vitro.
The conversion of [3H]estradiol to 2-hydroxyestradiol (2-OH-E2) by homogenates of porcine ovarian follicles was assayed in vitro in the presence and absence of 10 and 100 microM concentrations of the following potential substrates or inhibitors of estrogen-2/4-hydroxylase (E-2/4-H): (1) estrogens; estrone (E1), estriol (E3) and 17 alpha-estradiol (17 alpha-E2), (2) catecholestrogens; 2-hydroxyestradiol (2-OH-E2), 4-hydroxyestradiol (4-OH-E2) and 2-hydroxyestrone (2-OH-E1); (3) 2-methoxyestradiol (2-MeO-E2); (4) halogenated estrogens; 2-bromoestradiol, (2-Bromo-E2) 4-bromoestradiol and 2,4-dibromoestradiol; (5) androgens; testosterone (T), dihydrotestosterone (DHT) and androstenedione; (6) progesterone; (7) epinephrine; (8) inhibitors of steroid aromatase; aminoglutethimide and 4-hydroxyandrostenedione and (9) SKF 525A, an inhibitor of cytochrome P-450. Progesterone and 2-Bromo-E2 were the two most effective inhibitors (2-OH-E2 formation = 4 and 5% of control at 100 microM and 29.6 and 17.4% at 10 microM of progesterone and 2-Bromo-E2, respectively). 2-MeO-E2 at 100 microM was nearly as effective as progesterone in inhibiting E-2/4-H activity but only caused about 50% inhibition at 10 microM. The three catecholestrogens reduced 2-OH-E2 formation to about the same degree (21-23% of control at 100 microM). The 2,4-dibromo-E2 was equipotent with the catecholestrogens while 4-bromo-E2 was about half as effective. The phenolic estrogens, potential substrates for the enzyme, reduced 2-OH-E2 formation to different degrees, with E3 being the most effective. Among the androgens, DHT was almost as effective an inhibitor as the catecholestrogens, T was about half as effective while androstenedione had no effect. Epinephrine and the two inhibitors of aromatase did not inhibit E-2/4-H activity. SKF 525A inhibited E-2/4-H activity but with a potency only about 1/10th that reported for liver.
2-Methoxyestradiol↗
Peroxidatic catecholestrogen production by human breast cancer tissue in vitro.
The ability of breast cancer tissues from postmenopausal women to form catechol estrogens was examined by using a product isolation assay. Initial assays were carried out in the presence of either: (a) NADPH, the co-factor for monooxygenase mediated catecholestrogen (CE) formation or; (b) light-activated Tween 80 (LAT-80), a putative organic hydroperoxide co-factor for peroxidatic activity. Under monooxygenase conditions, CE formation by homogenates of 10 tumors did not exceed that obtained with heat denatured tissue. In contrast, 17 of 20 tumors incubated with LAT-80 synthesized significant amounts of CE (8.5 +/- 1.17 2-hydroxyestradiol [2-OH-E2] and 12.8 +/- 2.4 nmol/g protein/10 min 4-hydroxyestradiol [4-OH-E2]). Substitution of cumene hydroperoxide, an organic hydroperoxide, for LAT-80 enhanced estrogen 2/4 hydroxylase (E-2/4-H) activity over 200-fold, making it possible to characterize systematically the peroxidatic activity. The properties of peroxidatic E-2/4-H activity were similar to those of soluble peroxidases isolated from brain, including an acidic pH optimum, localization in the soluble fraction, an apparent Km in the range of 80 microM and an apparent Vmax in the range of 4000 nmol/g/protein/10 min for both 2- and 4-OH-E2. Under optimal assay conditions, peroxidatic E-2/4-H activity was identified in 10 of 13 tumors (2480 +/- 580 nmol/g protein/10 min for 2-OH-E2 and 2790 +/- 600 for 4-OH-E2). The level of activity detected suggests a biological relevance for CE formation by breast cancer tissue.
Animals↗
In-vitro and in-vivo potency of five new fluoroquinolones against anaerobic bacteria.
The MICs of difloxacin (A-56619), A-56620 and ofloxacin were similar or within one to two-fold dilutions against a variety of anaerobic bacteria. Ciprofloxacin was slightly less active (two- to four-fold dilutions) than difloxacin, A-56620 and ofloxacin. Norfloxacin was less active than the other fluoroquinolones tested against anaerobic bacteria. The MICs of norfloxacin, A-56620, ciprofloxacin and ofloxacin against most anaerobes, except the Gram-positive cocci, were lower at pH 8.1 than at pH 6.6. MICs of the fluoroquinolones against anaerobic cocci were the same at pH 6.6, 7.3 and 8.1. Norfloxacin and ciprofloxacin were most affected by the acidic pH. Ofloxacin and A-56620 were affected to a lesser extent by an acidic pH than norfloxacin and ciprofloxacin. The potency of difloxacin was similar at the three pHs tested. A subcutaneous anaerobic infection model in mice was used to determine the in-vivo efficacy of the new fluoroquinolones against Bacteroides fragilis. Difloxacin was the most potent compound in this test.
Anti-Bacterial Agents↗
Catechol estrogen formation by the CNS: regional distribution of estrogen-2/4-hydroxylase activity in rat brain.
Estrogen-2/4-hydroxylase (E-2/4-H) activity was measured by a direct product isolation assay in punch biopsy specimens obtained from nine nuclear regions from forebrain of adult male rats. Tritiated catechol estrogens were isolated from incubations of tissues with [6,7-3H]estradiol from all regions studied. The amount of 4-hydroxyestradiol (4-OH-E2) formed equaled or exceeded that of 2-hydroxyestradiol (2-OH-E2). There were significant regional differences in the amounts of catechol estrogen produced. The difference was nearly 8-fold between the arcuate-median eminence (ARC-ME) and the medial preoptic nucleus (POM), regions with the highest and lowest specific activities, respectively (37.7 +/- 6.2 vs. 5.1 +/- 0.7 pmol/mg protein/10 min 2-OH-E2, mean + SEM, n = 6). The supraoptic nucleus was the site of second highest concentrations of E-2/4-H activity (20.3 pmol 2-OH-E2/mg protein/10 min). Estrogen-2/4-H activity in the paraventricular (PVN) and periventricular (PERI) nuclear regions, though only about half that in the SON, was significantly greater than in the remaining brain areas (nucleus interstitialis striae terminalis, caudate, anterior hypothalamic and medial preoptic nuclei and cortex. The ARC-ME, the region with the highest E-2/4-H activity is where the dopaminergic neurones and terminals from the Gn-RH neurons are concentrated. The functions regulated by these two classes of neurones, the secretion of prolactin and gonadotrophins, respectively, have been the subject of most of the previous studies aimed at establishing the role of catechol estrogen formation in the hypothalamus.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Catecholestrogen production by porcine ovarian cells.
Catecholestrogens, formed locally, have been proposed to serve as local regulators of ovarian function. However, to date little or no activity of the critical biosynthetic enzyme, estrogen-2/4-hydroxylase, has been identified in the ovary. In the present study we have established the presence of this enzyme in porcine ovarian cells and characterized some of its biochemical features as well as its intraovarian distribution during the reproductive cycle. Homogenates of ovarian follicular tissue converted [3H]-17 beta-estradiol to the catecholestrogens 2-and 4-hydroxyestradiol (2-OH-E2 and 4-OH-E2) in a linear fashion for up to 60 min, with protein concentrations of equal to or less than 1 mg. The principal product, 2-OH-E2, was stable under the conditions of the assay. The reaction exhibited a dependence on nicotinamide cofactors, a pH optimum of 7.8, and an apparent Michaelis constant (Km) of approximately 10 microM for the production of 2-OH-E2 and 4-OH-E2. The activity of different ovarian preparations varied dramatically as a function of the reproductive cycle. Assayed at saturating substrate concentrations, immature follicular tissue and luteal tissue produced 50 or less pmol 2-OH-E2/mg protein X 40 min, while preovulatory follicles produced approximately 600 pmol 2-OH-E2/mg protein X 40 min. Even within the population of large presumptively preovulatory follicles, a variation in activity of more than 10-fold was encountered. Estrogen-2/4-hydroxylase activity of large preovulatory follicles correlated with the concentration of 17 beta-estradiol in the same follicles (r = +0.89, P less than 0.001). In large preovulatory follicles enzyme activity was present in both granulosa and theca layers. However, approximately 80% of the follicular activity was localized in the membrana granulosa. Under all conditions tested and in all ovarian compartments the formation of 2-OH-E2 was favored over that of 4-OH-E2. These studies show, for the first time, significant estrogen-2/4-hydroxylase enzyme activity within ovarian tissue. The striking increase in activity in the preovulatory follicle suggests physiological control of catecholestrogen synthesis. In conjunction with other data demonstrating stimulatory actions of catecholestrogens on ovarian cells, these observations are consistent with an intraovarian autocrine or paracrine regulatory function for these metabolites.
Animals↗
Purification and characterization of estrogen-2/4-hydroxylase activity from rabbit hypothalami: peroxidase-mediated catechol estrogen formation.
Estrogen-2/4-hydroxylase (E-2/4-H) activity of rabbit hypothalamic tissue was previously found to be localized in the soluble subcellular fraction. In the present study, the enzymatic activity responsible for catechol estrogen formation in this subcellular fraction of the rabbit hypothalamus was purified by ammonium sulfate fractionation, ion exchange chromatography, and chromatofocusing. E-2/4-H activity was found to be associated with a group of hemoproteins with peroxidase activity. The characteristics of this hypothalamic E-2/4-H activity were reestablished in light of a peroxidatic mechanism for catechol estrogen formation. Organic hydroperoxides stimulated E-2/4-H activity, presumably by serving as oxidizing cosubstrate required for peroxidase-mediated reactions. E-2/4-H activity in a 17,500 X g supernatant of hypothalamic tissue was linear with time for at least 10 min and with protein concentration to at least 100 micrograms/150 microliter. It displayed a pH optimum of 6 and an apparent Michaelis-Menten constant (Km) of 32 microM with respect to estradiol. The amounts of 4-hydroxyestradiol formed were comparable to those of 2-hydroxyestradiol. The characteristics established in this study for the peroxidase-type E-2/4-H were distinct from those of the particulate, NADPH-dependent 2-hydroxylases found in rat liver and in porcine blastocyst and ovary. These differences provide a basis for differentiating between the two types of enzymatic activity that can lead to catechol estrogen formation in vitro.
Animals↗
[Na-K]ATPase activity in proximal and distal tubules of the rat kidney: modification and application of a quantitative cytochemical technique.
A modified cytochemical assay for [Na-K]ATPase in cryostat sections of kidney was further characterized and used to quantify activity in seven functionally distinct sites along the rat nephron. The activity of [Na-K]ATPase was defined as the difference in ATPase activity in specifically identified tubules contained in serial sections incubated with and without ouabain. Preincubation of sections with ouabain was required for maximal inhibition of [Na-K]ATPase activity in several distal sites. The concentration of ouabain necessary for maximal inhibition of activity was 3.0 mM and half-maximal inhibition was obtained in all regions with 30-100 microM ouabain. In distal sites, [Na-K]ATPase formed a higher proportion of total ATPase activity (60-80 per cent) than in proximal sites (20-40 per cent). Enzyme activity was quantified using two different methods. The first measured activity over the basal region of tubules and gave an index of the concentration of [Na-K]ATPase over the basal lateral infoldings of cells composing the tubule. The second read activity over the entire cross section of tubules and provided an estimate of [Na-K]ATPase per length of tubule. The highest activities over the basal basal region were obtained from tubules of the distal nephron including the inner (MALin) and outer (MALout) medullary ascending limb, distal convoluted tubule (DCT) and connecting segment (CS). Lower activities were obtained in proximal convoluted (PCT) tubules, proximal straight (PS) tubules and the papillary collecting duct (PD). Distal convoluted tubules contained the highest activity per length of tubule. Other sites contained lower levels of activity in the following order: MALin greater than MALout greater than PCT greater than PD greater than PS. The modifications introduced increase the sensitivity and precision of this assay and permit the application of this technique to studies of [Na-K]ATPase activity in the major functional regions of the rat nephron.
Animals↗
Catechol estrogen formation by pig blastocysts during the preimplantation period: biochemical characterization of estrogen-2/4-hydroxylase and correlation with aromatase activity.
Formation of the catechol estrogens 2- and 4-hydroxyestradiol (2-OHE2 and 4-OHE2) from estradiol by pig blastocysts was studied using a direct product isolation assay for estrogen-2/4-hydroxylase (E-2/4-H). Blastocyst E-2/4-H activity was characterized biochemically using homogenates of blastocysts obtained on day 12 of pregnancy. This information was used to establish appropriate incubation conditions for the assay of E-2/4-H activity in blastocysts during the preimplantation period. Catechol estrogen formation was linear with time for up to 30 min and with blastocyst protein concentrations of up to 100 micrograms in a reaction volume of 150 microliters. The E-2/4-H activity of pig blastocysts was maximal at pH 7.9 and was not affected by the nonionic detergent Tween-80. The E-2/4-H activity was dependent on nicotinamide cofactor, with NADPH preferred over NADH for 2-OHE2 formation. The predominant catechol estrogen formed was 2-OHE2: maximum velocities (Vmax) for the formation of 2- and 4-OHE2 were 1570 and 174 pmol/mg protein . 30 min, respectively. The apparent Km values with respect to estradiol for 2- and 4-OHE2 were similar, 4.39 and 4.27 microM, respectively. Blastocyst E-2/4-H activity was detectable in one of two samples of blastocysts from day 10 of pregnancy (4.4 pmol 2-OHE2/mg protein . 30 min), increased to a maximum on days 12 and 13 (628 +/- 153 and 516 +/- 227 pmol 2-OHE2/mg protein . 30 min, respectively), and declined by day 14 (63.2 +/- 32.9 pmol 2-OHE2/mg protein . 30 min). The activity of E-2/4-H was positively correlated with aromatase activity assayed in the same tissue samples from days 10-14 of pregnancy. The surge in E-2/4-H activity coincides with several of the critical events that occur near the time of implantation. Our findings are consistent with the hypothesis that catechol estrogens mediate some of the actions of estrogens in early pregnancy in the pig.
Animals↗
The relationship between reproductive performance and blood pressure in the spontaneously hypertensive rat.
Blood pressure was measured serially throughout pregnancy in spontaneously hypertensive rats and in normotensive control rats of the Wistar-Kyoto strain. Changes in blood pressure were also correlated with the outcome of pregnancy. In control rats there was a small but significant decrease in blood pressure between days 20 and 22 of pregnancy (day of birth = day 22.5). In the spontaneously hypertensive rat this decrease occurred earlier, between days 18 and 19 of pregnancy, and was of greater magnitude. There was a positive correlation in the spontaneously hypertensive rat between the number of live-born pups and the magnitude of the decrease in blood pressure. Perinatal mortality, but not litter size, was greater in the spontaneously hypertensive rat than in control rats. Thus the physiologic mechanisms responsible for the decrease in blood pressure in the normal rat are preserved in the spontaneously hypertensive rat, and the successful completion and outcome of pregnancy depend on the capacity of the hypertensive rat to amplify these processes. Consequently, the spontaneously hypertensive rat is not an appropriate experimental model for hypertension of human pregnancy, in particular for preeclampsia, in which the presence of the conceptus characteristically causes blood pressure to rise, especially during the last trimester. However, an investigation of the factors responsible for the profound antihypertensive effect of pregnancy in rats could provide new insights into the mechanisms by which blood pressure is regulated during pregnancy and suggest new therapeutic approaches.
Animals↗
Pathways of hydrogen utilization from NADPH generated by glucose-6-phosphate dehydrogenase in circumventricular organs and the hypothalamo-neurohypophysial system: a cytochemical study.
Cytochemistry was used to examine the distribution of two pathways of utilization of hydrogen (Type I and Type II H) generated by glucose-6-phosphate dehydrogenase (G6PD) in circumventricular organs (CVOs) and the hypothalamo-neurohypophysial system in cryostat sections of rat brain. Type I H is defined as that portion of the total reducing equivalents (Total H) that is passed, in the intact cells, along the cytochrome chain (NADPH-diaphorase system). In the liver, energy from Type I H is used for cytochrome P-450-dependent oxidation of steroids, as well as xenobiotics. We proposed that mixed function oxidation, and therefore Type I H, would be preferentially localized in brain regions lacking a blood-brain barrier, such as CVOs and magnocellular cells with terminals in such brain regions. Type I H was identified in tissue sections using neotetrazolium. This reagent, when reduced, precipitates as formazan granules that can be quantified. The large difference in redox potential between NADPH and neotetrazolium ensures that only hydrogen (Type I H) passed in the intact cell along the cytochrome chain, can reduce the tetrazole. Total NADPH generation (Total H) from glucose-6-phosphate, was identified using medium containing phenazine methosulphate, a hydrogen acceptor that transfers all reducing equivalents from NADPH to the tetrazole. Type II H, the difference between Total and Type I H, is presumed to be used for NADPH-dependent biosynthetic functions such as lipid synthesis, or reduction of glutathione. In CVOs formazan granules indicative of Type I H were selectively concentrated and localized within cells throughout the SFO, organum vasculosum of the lamina terminalis, pineal gland and in the apical cytoplasm of columnar ependymocytes in the subcommissural organ. Formazan granules attributable to Type I H were also prominent throughout the hypothalamo-neurohypophysial system. Reaction product was present in the cytoplasm of some magnocellular neurons in both the supraoptic and paraventricular nuclei, in the median eminence, including the zona interna, and in and between cells in the neurohypophysis. The distribution of NADPH-diaphorase in sections incubated with NADPH instead of glucose-6-phosphate was similar to that of Type I H. These findings are consistent with the hypothesis that mixed function oxidation involving NADPH and the cytochrome chain occur in these brain regions.(ABSTRACT TRUNCATED AT 400 WORDS)
Animals↗
Differential effects of maternal stress on circulating levels of corticosterone, progesterone, and testosterone in male and female rat fetuses and their mothers.
Testosterone, progesterone, and corticosterone titers were measured by RIA in plasma of stressed and control pregnant rats and their male and female fetuses on days 17, 18, 19, and 21 of gestation and on the day of birth. The regimen of stress used (three 45-min periods of restraint under intense illumination daily from days 14-21 of pregnancy) causes failure of masculinization and defeminization of behavioral potentials in male offspring. In fetuses of both sexes, corticosterone titers increased sharply between days 17 and 18 postconception (pc) to a peak that was maintained through day 19 and then declined. This pattern resembled that obtained for testosterone in control male fetuses in which the levels of testosterone also rose sharply between days 17 and 18 pc. Corticosterone titers were elevated in samples obtained during the middle of the stress session from both the mothers (serum) and their male and female fetuses (plasma). Increased corticosterone levels were no longer evident in samples obtained from fetuses 75-165 min after the end of a stress session. Testosterone titers were altered by stress only in male fetuses. Their testosterone levels were elevated on day 17 pc, and the surge on days 18 and 19 pc, characteristic of control males, was absent in samples obtained 75-165 min after termination of stress. Progesterone titers were not affected by stress in either mothers or their fetuses. In both stressed and control groups, progesterone concentrations were identical in male and female fetuses, were higher in mothers than in fetuses, and declined in both fetuses and mothers between days 19 and 21 pc. Thus, a persistent effect of stress was observed only on testosterone and only in males.
Animals↗
Courtship stimulates aromatase activity in preoptic area of brain in male ring doves.
Aromatase activity was quantified in vitro in different regions of the basal forebrain of male ring doves at various times after the initiation of courtship. By 24 h after the introduction of a female dove into the cage of a male, there was a significant, nearly two-fold, increase in aromatase activity which was confined to the ventromedial preoptic area (VM-POA) where enzyme activity was concentrated. This increase in aromatase activity, which was sustained through day 5 of courtship, presumably serves to activate nest-oriented behavior.
Animals↗
Spenolimycin, a new spectinomycin-type antibiotic. III. Biological properties.
Spenolimycin is a new spectinomycin-type antibiotic isolated from Streptomyces gilvospiralis sp. nov. In vitro, it was active against a wide variety of aerobic Gram-positive and Gram-negative bacteria and Neisseria gonorrhoeae. It was two to four-fold more active against N. gonorrhoeae than spectinomycin. Spenolimycin was effective in the standard mouse protection test against Escherichia coli, Klebsiella pneumoniae and Streptococcus pneumoniae.
Animals↗
Steroid structure and function--IX. Molecular conformation of catechol estrogens.
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Estrogens, Catechol↗
Environmental stress alters the developmental pattern of delta 5-3 beta-hydroxysteroid dehydrogenase activity in Leydig cells of fetal rats: a quantitative cytochemical study.
Quantitative cytochemistry was used to determine the effect of subjecting pregnant rats to environmental stress on the activity of delta 5-3 beta hydroxysteroid dehydrogenase (3 beta-HSD) in Leydig cells of their fetuses. Enzyme activity was measured by microspectrophotometry in individual Leydig cells in cryostat sections of fetal testes on Days 16-21 postconception. Fetuses of stressed mothers lacked the peak of enzyme activity on Days 18 and 19 of gestation that is characteristic of Leydig cells of normal fetuses at this time. In addition, both before and after these 2 days, 3 beta-HSD activity in Leydig cells of stressed fetuses was significantly higher than normal. The altered developmental pattern of 3 beta-HSD activity in the stressed fetuses largely corresponds to the changes in plasma testosterone found previously in male fetuses of mothers exposed to the same regimen of stress. Thus, in the fetal Leydig cell, the activity of 3 beta-HSD, a key steroidogenic enzyme, can be modified by environmental stress, and provides an index of steroidogenic activity of the fetal testes and of the titers of circulating testosterone.
3-Hydroxysteroid Dehydrogenases↗
Influence of maternal stress on the developmental pattern of the steroidogenic function in Leydig cells and steroid aromatase activity in the brain of rat fetuses.
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3-Hydroxysteroid Dehydrogenases↗