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Biomedical subjects

M E Ortiz

Publications and source records attributed to M E Ortiz.

At least 19 recordsLinked to original sources

Cytologic, hormonal, and ultrasonographic correlates of the menstrual cycle of the New World monkey Cebus apella.

Few reports on the reproductive physiology of Cebus apella have been published. In this study we characterized menstrual cycle events by means of vaginal cytology, ultrasonography (US), and hormonal measurements in serum during three consecutive cycles in 10 females, and assessed the probability that ovulation would occur in the same ovary in consecutive cycles in 18 females. The lengths and phases of the cycles were determined according to vaginal cytology. Taking the first day of endometrial bleeding as the first day of the cycle, the mean cycle length +/- SEM was 19.5+/-0.4 days, with follicular and luteal phases lasting 8.2+/-0.2 and 11.3+/-0.4 days, respectively. The follicular phase included menstruation and the periovulatory period, which was characterized by the presence of a large number of superficial eosinophilic cells in the vaginal smear. The myometrium, endometrium, and ovaries were clearly distinguished on US examination. During each menstrual cycle a single follicle was recruited at random from either ovary. The follicle grew from 3 mm to a maximum diameter of 8-9 mm over the course of 8 days, in association with increasing estradiol (E(2)) serum levels (from 489+/-41 to 1600+/-92 pmol/L). At ovulation, the mean diameter of the dominant follicle usually decreased by >20%, 1 day after the maximum E(2) level was reached. Ovulation was associated with an abrupt fall in E(2), a decreased number of eosinophilic cells, the presence of leukocytes and intermediate cells in the vaginal smear, and a progressive increase in progesterone (P) levels that reached a maximum of 892+/-65 nmol/L on days 3-6 of the luteal phase. The menstrual cycle of Cebus apella differs in several temporal and quantitative aspects from that in humans and Old World primates, but it exhibits the same correlations between ovarian endocrine and morphologic parameters.

Animals↗

Post-coital administration of levonorgestrel does not interfere with post-fertilization events in the new-world monkey Cebus apella.

BACKGROUND: Experimental evidence to disprove the belief that emergency contraception with levonorgestrel (LNG) prevents pregnancy by interfering with post-fertilization events is lacking. Here we determined the effect of post-coital and pre-ovulatory administration of LNG on fertility and ovulation, respectively, in the Cebus monkey. METHODS: To determine the effect on fertility, LNG 0.75 mg or vehicle were administered orally or s.c. once or twice within the first 24 h after mating occurring very close to the time of ovulation. Females that became pregnant were aborted with mifepristone and re-entered the study after a resting cycle until each of 12 females had contributed, in a randomized order, two LNG and two vehicle-treated cycles. To determine the effect on ovulation, LNG 0.75 mg or vehicle were injected twice coinciding with follicles smaller or larger than 5 mm in diameter. Six females contributed five treated cycles each. RESULTS: The pregnancy rate was identical in vehicle- and LNG-treated cycles. LNG inhibited or delayed ovulation only when treatment coincided with a follicle <5 mm diameter. CONCLUSION: In Cebus monkeys, LNG can inhibit or delay ovulation but, once fertilization has taken place, it cannot prevent the establishment of pregnancy. These findings do not support the hypothesis that emergency contraception with LNG prevents pregnancy by interfering with post-fertilization events.

Animals↗

Oxidation of Hantzsch 1,4-dihydropyridines of pharmacological significance by electrogenerated superoxide.

PURPOSE: To study the reaction of a series of Hantzsch dihydropyridines with pharmacological significance such as, nifedipine, nitrendipine, nisoldipine, nimodipine, isradipine and felodipine, with electrogenerated superoxide in order to identify products and postulate a mechanism. METHODS: The final pyridine derivatives were separated and identified by gas chromatography/mass spectrometry (GC-MS). The intermediates, anion dihydropyridine and the HO2*/HO2- species, were observed from voltammetric studies and controlled potential electrolysis was used to electrogenerate O2*-. RESULTS: The current work reveals that electrogenerated superoxide can quantitatively oxidize Hantzsch dihydropyridines to produce the corresponding aromatized pyridine derivatives. CONCLUSIONS: Our results indicate that the aromatization of Hantzsch dihydropyridines by superoxide is initiated by proton transfer from the N1-position on the 1,4-dihydropyridine ring to give the corresponding anion dihydropyridine, which readily undergoes further homogeneous oxidations to provide the final aromatized products. The oxidation of the anionic species of the dihydropyridine is more easily oxidized than the parent compound.

Calcium Channel Blockers↗

Trypanosoma cruzi: inhibition of parasite growth and respiration by oxazolo(thiazolo)pyridine derivatives and its relationship to redox potential and lipophilicity.

Chagas' disease constitutes a therapeutic challenge because presently available drugs have wide toxicity to the host and are generally ineffective in the chronic stages of the disease. A series of oxazolo(thiazolo)pyridene derivatives were studied on Trypanosoma cruzi epimastigote growth and oxygen consumption and their electrochemical (redox) potentials and lipophilicity. The derivatives produced different degrees of parasite growth and respiration inhibition on CL Brener, LQ, and Tulahuen strains of T. cruzi epimastigotes. Respiratory chain inhibition appears to be a determinant of the trypanosomicidal activity of these compounds, since a significant correlation between respiration and culture growth inhibition was found. A similar correlation was found, within the different structural subfamilies, between toxic effects and the ability of the compounds to be oxidized in aqueous media. The inhibition of respiration and of parasite growth in culture increases with the lipophilicity of the substituents on the oxazolopyridine nucleus. No difference in the action of these derivatives was found among the different parasite strains. It is concluded that these compounds may have a potential usefulness in the treatment of Chagas' disease.

Animals↗

Effect of nitric oxide synthase inhibitors on ovum transport and oviductal smooth muscle activity in the rat oviduct.

The effect of the inhibition of nitric oxide synthase (NOS) on ovum transport and oviductal motility in rats was investigated. Three different NOS inhibitors were injected into the ovarian bursa at oestrus or day 3 of pregnancy. Oviducts and uteri were flushed 24 h later and the presence of ova was recorded. In oestrous and pregnant rats, treatment resulted in accelerated egg transport, as shown by a decrease in the number of ova present in the oviducts. In cyclic rats, intrabursal injection of 1 mg kg-1 of either N-monomethyl-L-arginine (L-NMMA) or N omega nitro-L-arginine methyl ester (L-NAME) elicited a 30% reduction in the number of ova present in the oviducts, whereas in pregnant animals, the same dose of L-NMMA produced a reduction of 40%. Simultaneous administration of the NO donor spermine NONOate (5 mg kg-1) completely reversed the effect of L-NMMA. Tubal motility was assessed by microsphere displacement analysis within the oviduct. Surrogate ova were transferred to the oviductal lumen at oestrus and 24 h later the effect of intraoviductal injection of 1 microgram L-NMMA or vehicle was assessed. The microspheres in the isthmus showed an oscillating motion, and periods in which movement was not detectable. However, L-NMMA treatment produced a 3.6-fold increase in the maximum instant velocities and a significant reduction in the resting periods of the microspheres compared with the control group (P < 0.001). These results provide evidence that NO inhibition increases tubal motility that results in accelerated ovum transport, and indicate that NO could act as a paracrine signal between different layers of the oviductal wall, providing a role for endogenous NO in regulation of tubal function.

Analysis of Variance↗

Sperm migration into and through the oviduct following artificial insemination at different stages of the estrous cycle in the rat.

In order to examine whether sperm migration into and through the oviduct follows an invariable pattern or is subject to regulation, rats in proestrus, estrus, metestrus, or diestrus were inseminated in the upper third of each uterine horn with 10-20 million epididymal spermatozoa. Three or eight hours later, the numbers of spermatozoa free and adhering to the epithelium in the ampullary and isthmic segments were determined. A significantly higher number of spermatozoa were recovered in estrus than in other stages, at 3 h than at 8 h, and at all stages from the isthmus than from the ampulla. Spermatozoa adhering to the epithelium were observed only in proestrus and estrus and in the isthmus. The effect of exogenous estradiol-17beta (E2) and progesterone (P4) on sperm migration was investigated in rats in which the estrous cycle was inhibited pharmacologically. E2 facilitated sperm migration into the oviduct and P4 antagonized this effect, whereas P4 alone had no effect. Concomitant treatment with E2+P4 induced adhesion of spermatozoa to the oviductal epithelium. In conclusion, the pattern of sperm migration into and through the rat oviduct varies with the stage of the cycle, being dependent on E2 and P4. The adhesion of spermatozoa to the rat oviductal epithelium is stage- and segment-specific and requires the combined action of both hormones.

Animals↗

Modified Dakin's solution for cutaneous vibrio infections.

Vibrio species, specifically Vibrio vulnificus, are known to be endemic to warm saltwater environments. As a human pathogen they are capable of causing severe, progressive, necrotizing infections. The lesions are bullous in nature and often require wide surgical debridement due to the aggressiveness of this organism. The literature supports prophylactic antibiotic therapy for those with preexisting hepatic dysfunction or immunocompromise. The authors routinely implement prophylactic antibiotic coverage with doxycycline 100 mg every 12 hours for vibrio in patients with wounds exposed to or acquired in saltwater. In addition, they institute topical therapy with 0.025% sodium hypochlorite solution (modified Dakin's), based on their in vitro study of vibrio sensitivity to antimicrobials. Over the past 2 years, the authors have treated 10 patients with this protocol for cutaneous vibrio infections confirmed by quantitative cultures. None of these patients experienced progression of infection requiring operative debridement-contrary to the aggressive nature of this organism documented in other reports.

Adolescent↗

The expression of alpha(v) and beta3 integrin subunits in the normal human Fallopian tube epithelium suggests the occurrence of a tubal implantation window.

The co-expression of alpha1beta1, alpha4beta1 and alphavbeta3 integrins in the human endometrium coincides with the implantation window. The alphavbeta3 integrin is expressed in the apical surface of the luminal epithelium and may serve to anchor trophoblast cells in the adhesion phase of implantation. Using immunohistochemistry, we compared the expression of alphav, alpha1, alpha4 and beta3 integrin subunits in samples of normal human Fallopian tube and endometrium obtained from five women in the non-receptive period (luteal phase days 2-4) and from another five women in the receptive period (luteal phase days 6-8). The staining was quantified visually on a scale of 0 to ++, according to the intensity and density of stained cells. The alphav subunit is expressed in the Fallopian tube epithelium during both periods in a pericellular distribution. The beta3 subunit is also expressed in the same location, but it is up-regulated during the period of endometrial receptivity. The other subunits are expressed in localizations which are not relevant to trophoblast adhesion and exhibit little or no difference in the level of expression between the non-receptive and receptive periods. Based on these results we postulate that the expression of the beta3 subunit in the human tubal epithelium is under the same systemic controlling signals as in the endometrium and that the normal tubal epithelium may have an implantation window, at about the same time as the endometrium, that affords the opportunity for trophoblast attachment should a 5-7 day embryo be unduly retained in the tube.

Antigens, CD↗

Electrochemical generation and reactivity of free radical redox intermediates from ortho- and meta-nitro substituted 1,4-dihydropyridines.

This paper reports a comprehensive study by cyclic voltammetry on the electrochemical characteristics and the reactivity of the one-electron reduction product from a series of nitro aryl 1,4-dihydropyridines in mixed and aprotic media. In addition, the effects of 1,4-DHP on the oxygen consumption of T. cruzi epimastigotes are reported. One-electron reduction products from 1,4-DHP derivatives significantly reacted with both thiol compounds and the nuclei acid bases, adenine and uracil. This reactivity was significantly higher than the natural decay of the radicals in mixed media. Based on these results the following tentative order of reactivity towards the xeno/endobiotics is as follows: cysteamine > glutathione > adenineuracil. Both the stability and the reactivity of the nitro radical anions electrochemically generated from 1,4-DHP showed a linear dependence with pH. The sensitivity to pH of the radicals derived from o-nitro substituted derivatives was significantly higher than m-nitro substituted derivatives. On the other hand, in all cases an increase of pH produced a significant decrease in the interaction rate constant. Interaction studies carried out in aprotic media did not show any reactivity of the radicals towards both thiol compounds and the nuclei acid bases, adenine and uracil. Therefore, we concluded that the interaction process requires certain proton activity in the media. All the tested 1,4-dihydropyridines inhibited the oxygen consumption by T. cruzi epimastigotes, Tulahuén strain. The drugs with higher electron-affinity produced greater inhibition than those with lower electron-affinity (i.e. nicardipine vs nifedipine).

Animals↗

Reactivity of the one-electron reduction product from nifedipine with relevant biological targets.

The reactivity of the electrochemically generated nitro radical anion from nifedipine, a nitro aryl 1,4-dihydropyridine derivative, with relevant endobiotics and thiol-containing xenobiotics, was quantitatively assessed by cyclic voltammetry. The method was based on the decrease in the return-to-forward peak current ratio after the addition of compounds. A quantitative procedure to calculate the respective interaction constants between the radicals and the xeno/endobiotics is also provided. In the optimal selected conditions, i.e. mixed media (0.015 M aqueous citrate/DMF: 40/60, 0.3 M KCl, 0.1 TBAI) at pH 9.0 the following order of reactivity was obtained: glutathione > uracil > adenine and cysteamine > N-acetylcysteine > captopril > penicillamine. In all cases, the interaction rate constants for these derivatives were greater than the natural decay constant of the radical. Studies on the reactivity at pH 7.4 were also conducted. Results from these experiments indicate a significant reactivity between the radical and the endo/xenobiotics. The increase in the stability of the radical anion by increasing the pH of the mixed media resulted in a decreased reaction with the endo/xenobiotics tested. Computerized simulation with DIGISIM 2.0 of the proposed mechanisms fitted very well with the experimental results for both the natural decay of the radical and its reaction with the tested compounds.

Calcium Channel Blockers↗

Mechanisms of action of intrauterine devices.

The major effect of all intrauterine devices (IUD) is to induce a local inflammatory reaction in the endometrium whose cellular and humoral components are released into the uterine cavity. This inflammatory reaction has a variable effect on the reproductive strategy of the species studied. For example, this foreign body reaction can be localized within the uterus of rodents; and in farm animals it can have striking extrauterine effects. Thus, the action of IUDs in humans cannot be discerned from animals. In humans, copper ions released from Cu-IUDs enhance the inflammatory response and reach concentrations in the luminal fluids of the genital tract that are toxic for spermatozoa and embryos. In women using the IUD, the entire genital tract seems affected, at least in part, because of luminal transmission of the fluids that accumulates in the uterine lumen. This affects the function or viability of gametes, decreasing the rate of fertilization and lowering the chances of survival of any embryo that may be formed, even before it reaches the uterus. Studies on the recovery of eggs from women using IUDs and from women not using contraception show that embryos are formed in the tubes of IUD users at a much lower rate compared with nonusers. This is believed to be the major action of IUDs. Therefore, the common belief that the major mechanism of action of IUDs in women is through destruction of embryos in the uterus (i.e., abortion) is not supported by the available evidence. In Cu-IUD users, it is likely that few spermatozoa reach the distal segment of the fallopian tube, those that encounter an egg may be in poor condition. Thus, the few eggs that are fertilized have little chance for development and their possibility for survival in the altered tubal milieu become worse as they approach the uterine cavity.

Animals↗

Increased sensitivity and accumulation of estradiol in the rat oviduct during early pregnancy.

We have previously reported that a single injection of estradiol-17 beta (E2) given on day 3 of pregnancy (P3) is far more effective for accelerating oviductal transport in the rat, than treatment given on day 1 (P1). In order to quantify this change, dose-response curves were established for six different doses of E2 (range 0.031 to 1.00 micrograms per animal) given on P1, P2 or P3. In addition, a possible mechanism was explored by comparing the plasmatic and oviductal levels of E2 between 30 and 180 min following treatment with E2 on P1 or P3. As the interval from ovulation to treatment was increased, the transport of a larger number of embryos was accelerated and a smaller dose was required. The minimal effective dose decreased 30-fold from P1 to P3, the oviducts accumulated 20% to 90% more E2 on P3 than on P1, tissue levels were 6- to 48-fold higher than plasma levels and the latter did not differ between P1 and P3. It is concluded that the oviduct exhibits increased sensitivity and responsiveness to E2 on P3 and this is associated with greater accumulation of the hormone in the organ, not attributable to higher E2 plasma levels.

Animals↗

Uterine kallikrein in the early pregnant rat.

Uterine homogenates of cycling and early pregnant Sprague Dawley rats and purified rat urinary kallikrein showed similar curves of displacement of 125I-kallikrein binding to a polyclonal antibody. Uterine kallikrein concentration measured by RIA was 8.7 +/- 2 SEM ng/g wet weight during the cycle (n = 6 in diestrus and metestrus) and 20.8 +/- 2 SEM (n = 7) ng/g wet weight on Day 7 of pregnancy (P7) (p < 0.001). On P7, kallikrein concentration was increased 12.4-fold in the implantation nodes, as compared to the interimplantation segments. Uterine homogenates of rats on P7, submitted to DEAE-cellulose chromatography and Sephadex gel filtration, yielded two fractions containing kallikrein immunoreactivity and kininogenase activity, with molecular masses that ranged from 120-125 kDa and 39-43 kDa, respectively. In the RIA, both fractions displayed parallelism with purified kallikrein. Enzymatic activity was expressed after activation by trypsin. It was inhibited by aprotinin, PMSF, p-amino-benzamidine, and leupeptin, but not by soybean or ovomucoid trypsin inhibitors. Kallikrein mRNA was demonstrated by reverse transcriptase/polymerase chain reaction in uteri of nonpregnant and P7 rats. These results show that rat uterus synthesizes one or more serine proteases that are immunologically and enzymatically related to tissue kallikrein in the implantation node on P7--determined both by an increment of whole uterus kallikrein content and a depletion of the interimplantation segments--suggests that kallikrein may play a role in the vasoactive changes of implantation.

Animals↗

Importance of uterine expulsion of embryos in the interceptive mechanism of postcoital oestradiol in rats.

This study compares the interceptive effectiveness of 1 microgram oestradiol given as a single s.c. injection at 0900 or 1700 hours on Day 1, 2, 3, or 4 of pregnancy in rats. Increasing the interval from ovulation to treatment accelerated the oviducal transport of a larger number of embryos, the majority of which were lost from the genital tract. However, after treatment at 1700 hours on Day 3 the majority of accelerated embryos were retained in the uterus. The number of implanted embryos on Day 14 was equal to the number of eggs remaining in the tract 24 h after treatment. As a consequence, the highest interceptive effectiveness was obtained with treatment given at 1700 hours on Day 2 and at 0900 hours on Day 3 of pregnancy. Accelerated oviducal transport and uterine expulsion of embryos begin to dissociate after Day 2 of pregnancy in the rat. This explains why the most effective treatments to accelerate oviducal transport are not always the most effective to reduce the number of implantations. These data emphasize the importance of retentive and expulsive properties of the uterus for fertility and infertility.

Animals↗

High potassium concentration and the cumulus corona oocyte complex stimulate the fertilizing capacity of human spermatozoa.

Progressively motile spermatozoa were incubated for 24 hours in culture media containing 4.7 or 25 mM K, in the presence or absence of hamster cumulus oophorus. The percentage of spermatozoa with progressive motility was significantly higher at 24 hours in the presence of cumulus corona oocyte complexes, irrespective of K concentration. A significant decrease in sperm mortality was observed with the association of 25 mM K and cumulus cells. A higher percentage of acrosome reaction was observed in spermatozoa incubated in 25 mM K when compared with 4.7 mM K, irrespective of time and the presence or absence of cumulus. The percentage of penetrated oocytes at 2 and 5 hours of incubation was higher when sperm had been incubated in 25 mM K than in 4.7 mM K. The presence of cumulus in the culture medium induced an additional significant increase in the percentage of penetrated oocyte. Although at 24 hours of incubation the percentage of acrosome reaction was higher than at 2 and 5 hours, the percentage of penetrated oocytes did not increase proportionally.

Acrosome↗

Embryos of different ages transferred to the rat oviduct enter the uterus at different times.

Indirect evidence of embryo signalling to the oviduct was sought in rats by examining the transport of embryos of different ages. One-cell or four-cell embryos were transferred to the oviducts of recipient rats on Day 1 of pregnancy, and the number, condition, and location of native and transferred embryos was assessed on Day 4. To control for the effect of the presence of foreign embryos and excess number of eggs and the transfer procedure upon the fate of native embryos, other groups of rats were sham-operated or left undisturbed. Recipients had a mean number of ova significantly higher than controls. In controls and recipients of 1-cell embryos, the majority of eggs reached the morula stage and all of them were located in the oviducts. In those animals receiving 4-cell embryos, half of the eggs had reached the blastocyst stage and 28% were in the uteri (p less than 0.005). These results support the idea that advanced embryos can influence the timing of their entrance to the uterus in rats.

Age Factors↗