Search

Differences in uterotonic activity were observed between zoapatle Montanoa (Cerv.), plants growing in their natural habitat and plants growing in an experimental agricultural plot. Details of an in vitro analogic model for assaying uterotonic potency in guinea pig strips is described. Important species differences on the uterine response to zoapatle aqueous crude extract were noticed in rats, hamsters, guinea pigs, cats and Rhesus monkeys. The need for proper biological evaluation of chemical substances already isolated from zoapatle specimens, is mentioned, and the advantages of working with zoapatle specimens grown under controlled ecological conditions are pointed out.An in vitro analogic method for comparing the uterotonic potency of dialysed zoapatle aqueous crude extract (DZ-ZACE) with oxytocin is described. Uterine strips from estrogenized guinea pigs were selected as the appropriate animal model after trials with estrogenized rats, hamsters, cats, and Rhesus monkeys. Results of the in vitro uterotonic effect produced by 21 different zoapatle Montanoa plants collected from different parts of Mexico and grown in an experimental agricultural field station indicate considerable difference, with less variation in uterotonic oxytocin/equivalents among the plants grown at the field station than in wild specimens. The general reduction in uterotonic potency of plants grown at the field station may be due to differences in the time of collection: wild specimens were collected in winter 1980, while field station specimens were collected just after flowering in early summer 1981. The findings clearly demonstrate the influence of the plant environment on the expression of biological response. Because of its lack of toxicity, abundance, ease of production, and the availability of an efficient analogic in vitro assay for the evaluation of relative uterotonic potency, ZACE may eventually be the source of a new agent to intercept pregnancy.

The changes in the activity and in the subcellular distribution of three lysosomal enzymes have been studied in the normal human endometrium during the menstrual cycle by the use of highly sensitive fluorimetric technics. Total activity of lysosomal enzymes increased during the maturation of the endometrial tissue from the proliferative to the secretory phase. This increase is greater (105%) for N-acetyl-β-D-glucosaminidase, than for acid phosphatase (77%) or β-glucuronidase (65%). During the proliferative phase the relative activity of the three enzymes found free in the cell cytoplasm was similar and ranged from 21% for N-acetyl-glucosaminidase to 32% for β-glucuronidase. The main difference found between the proliferative and the secretory phase of the menstrual cycle was a drastic increase -- both absolute and relative -- of the cytoplasmic free activity of the lysosomal enzymes. The increase in total free activity represented approximately two, three and four times the amount found free during the proliferative phase for β-glucuronidase, acid phosphatase and N-acetyl-glucosaminidase, respectively. In all cases the relative activity found free represented almost 50% of the total enzymatic activity. The membrane bound activity of the three enzymes also showed a significant increase during the secretory phase, although less dramatic than the free activity.

Zoapatle aqueous crude extract (ZACE) obtained from Montanoatomentosa (M.t.) inhibits the invitro spontaneous contractility pattern of rat uterine tissue. The opposite effect was observed with ZACE from Montanoafrutescens (M.f.) in the same preparation. Both plant extracts, M.t. and M.f., increased the invitro spontaneous contractility pattern in the uterine guinea pig assay. In depolarized uterine tissue, propanolol (β-blocker) inhibited the relaxing effect induced by M.t. Atropine (cholinergic antagonist) abolished the increase in uterus contractility produced by the presence of M.f.

The activities and subcellular distribution of three lysosomal enzymes have been studied on the endometriurn of three groups of women wearers of a progesterone releasing IUD, a Cu-T-200 and an inert T intrauterine devices. The changes in the parameters studied induced by the presence of an inert T were quantitatively small and of limited biological significance. The presence of a copper T induced significant changes mainly during the secretory phase: total activities of the lysosomal enzymes were at least doubled, and in the case of β-glucuronidase the value was almost four times that found in the inert T users. The more outstanding differences observed in the copper T users were in the amount of free activities of glucosaminidase, acid phosphatase and β-glucuronidase which were found to be more than 3, 4 and 6 times higher, respectively, than the activities found in the women using a placebo T. The results obtained in endometria exposed to the continuous release of progesterone were completely different from that observed in the endometria exposed to the continuous release of copper. In this case the existence of a remarkable stability during the menstrual cycle was readily apparent with small or no changes in the free activity of the enzymes during the secretory phase. These results are related to the mechanism of action of the different T-IUDs and to the observed changes in the patterns of bleeding and pain that are observed in women wearers of these contraceptive devices.

Zoapatle aqueous crude extract (ZACE) from Montanoa tomentosa has been used as an oral contraceptive in traditional Mexican medicine for centuries. Kaurenoic acid, as isolated from Montanoa frutescens, and the methyl esters of 15-hydroxy-dihydro-kaurenoic acid and 15-keto-dihydro-kaurenoic acid were tested in vitro on human sperm motility and viability. The estimated ED50 concentrations immobilization were 374, 126 and 58 micrograms/ml, respectively using 15 X 10(6) sperms/500 microliter. Kaurenoic acid and both of its derivatives displayed only weak to negligible capacity for killing human sperms.The effects on human sperm motility and viability of 3 compounds isolated from an aqueous extract of Montanoa frutescens Cerv (Compositae, Heliantheae) were tested. The plant is related to the zoapatle, traditionally used as a contraceptive, and known to have uterotonic and luteolytic activity. Kaurenoic acid, its 15 hydroxy- and its methyl ester were prepared from the polar fraction, and dissolved in 95% ethanol at a concentration of 10 mg/ml. Human sperm, suspended in Bigger's Whitten and Wittingham medium at a final concentration of 15 million sperm per 500 mcl were incubated at 37 degrees C. for 60 seconds. Judging sperm motility under phase contrast microscopy, kaurenoic acid was 0.89 times less effective, and the hydroxy kaurenoic acid 2.6 times more effective than verapamil, while the keto derivative was 5.7 times more potent than verapamil at the same concentration. Sperm viability assessed by vital staining was hampered only about 10% by the keto-kaurenoic acid, 21% by kaurenoic acid, and 33% by hydroxy-kaurenoic acid. Doses required for immobilization of 50% of human sperms ranged from 58 to 374 mcg/ml. This is 225 times lower than that reported for gossypol in vitro, and much lower than the millimolar range needed to inactivate sperm with nonoxynol. Thus these drugs may prompt a search for analogs useful in vaginal contraceptives.

Intrauterine administration of zoapatle aqueous crude extract (ZACE) from Montanoafrutescens on the fourth day of rat pregnancy, at concentrations equivalent to 50 mg and 5 mg of dry leaves, was associated with total inhibition of implantation sites. On the other hand, ZACE from Montanoatomentosa equivalent to 50 and/or 100 mg of dry leaves, prepared and administered in the same fashion, did not inhibit the number of implants by day 11 of pregnancy. However, most implants were found abnormal, of blue color, poor orientation or spacing; these morphological changes could be considered as reabsorption sites.

Intrauterine administration of Zoapatle aqueous crude extract (ZACE) prepared from Montanoatomentosa s.s.p. tomentosa, on day 4 of pregnancy, did not alter the normal uterine morphological changes by days 5 and 8 of pregnancy in the rat. On the other hand, ZACE made from Montanoafrutescens caused profound alterations on the uterine structures, such as: focal loss of epithelial lining, thickened blood vessels and alterations in stroma cells in rat endometrium. Morphological alterations correlate well with the antiimplantation effect associated with the intrauterine administration of these plant extracts. Scanning electron micrographs of leaves from Montanoatomentosa and Montanoafrutescens revealed striking differences in glandular secretion and trichomes.

Crossed-perfusions of homologous loci of the mesencephalic reticular formation were carried out on pairs of “encephale isole” cats in order to detect the possible secretion of a chemical substance mediating electrocortical synchronization and desynchronization. Special care was taken in quantitatively determining (i) the precise site and diffusion of perfusates in different brain stem structures by scintillographic radioactivity determination of a 14C-labeled cerebrospinal fluid used as perfusate solution; (ii) the spontaneous variations in the amount of electroencephalogram (EEG) synchrony by determining changes in concentration of delta EEG waves and amplitude of cortical recruiting responses; and (iii) possible variations in hydrodynamic function of the perfusion system reflected in changes in body temperature, electrocardiogram rate, and blood pressure. The results showed that perfusion of the tegmental mesencephalic reticular formation (with maximal radioactivity in the ipsilateral formation) with perfusate with possible desynchronizing properties significantly reduced the concentration of delta waves and the amplitude of recruiting responses under stable conditions of temperature, heart rate, and blood pressure.

The effect of zoapatle aqueous crude extract (ZACE) was further studied and partially characterized upon human and rabbit spermatozoa. ZACE prepared from Montanoa tomentosa s.s.p tomentosa did not influence sperm motility or viability in a wide range of ZACE concentrations tested; on the other hand, ZACE prepared from Montanoa frutescens had immediate and constant inhibitory effect upon motility and decreased cell viability. Red cell lysis was readily observed with ZACE-frutescens, but not with ZACE-tomentosa. The effect of time on ZACE-frutescens potency for inducing red cell lysis was observed.

Increase in menstrual blood loss (MBL) is one of the main causes for discontinuing use of intrauterine devices (IUDs). Increased blood loss may be especially important in the developing countries where some women have pre-existent anemia and nutritional deficiencies. MBL and hemoglobin (Hb) were measured in a group of 78 women prior to the insertion of one of the following eight devices: Lippes Loop D, 11 subjects; TCu-200, 14; TCu-380B, 12; TCu-220C, 8; UPS-40, 7; UPS-65, 10; norethisterone (10 ug/day) and levonorgestrel (2 ug/day), 9 and 7, respectively. Based on changes in the MBL, the women could be divided into two groups. One wearing the inert and the copper-releasing devices which increased MBL from between 43.6 and 21.85 ml when compared to their own control cycle and decreased Hb between 0.16 and 1.19 gm; and one wearing steroid-releasing devices associated with no change in MBL or a decrease of MBL up to 25.7 ml and no significant changes in Hb concentration.

The fibroblast, a widely distributed cell of mesenchymal origin, is the most abundant cell type of loose connective tissue (1). Although these cells participate in many functions, their name refers to their ability to elaborate connective tissue fibers. The fibroblast's major activity is exhibited in the formation of new connective tissue components during growth, regeneration and repair (2). These cells have been shown, by Dougherty et al., to play a major part during the development of inflammatory reactions (3-5). Fibroblasts are extremely sensitive target cells to anti-inflammatory steroids (6-8). The growth inhibitory effects of anti-inflammatory steroids on conective tissue cells in vitro (6-10) and in vivo (11, 12) is a well-known phenomenon. The following is a description of the early morphological changes occurring in tissue culture fibroblasts, in the presence of the most potent naturally occurring steroid, Cortisol, and the most potent topical synthetic anti-inflammatory steroid, fluocinolone acetonide (13-15).

In a previous study EEG abnormalities were detected in 2 of 24 healthy women during the 1st 3 months of treatment with various ovulatory and anovulatory contraceptive regimens. The present study is a follow up of the same group of women to detect possible EEG abnormalities occurring at the 6th and 12th months of treatment and those occurring within the 1st month after discontinuation of contraception. Only 17 women were studied (the 7 others were excluded due to voluntary treatment termination and pregnancy); they were divided into groups: 1) coil (5 cases) 2) microdoses (5 cases) and 3) sequential (4 cases) and combined (3 cases). In each case 3 consecutive pairs of routine EEG recordings (during the 6th and 12th months of contraceptive treatment and within 1st month of contraceptive discontinuation) and an equal number of plasma samples for progesterone/17 beta estradiol determination were taken. EEG routine recordings were also practiced during menstruation and during sleep. Procedures for routine EEG recordings and interpretation hormone determination standardization of dates and statistical significance of changes along the menstrual cycle were done as in the previous study. 4 of 7 patients in group 3 (the anovulatory sequential and combined contraceptive regimen) exhibited subclinical pseudoparoxysmal EEG discharges at the 6th and 12th months of treatment. These abnormalities may be due to the effect of specific hormone content of these particular compounds as they were not seen in control records taken before and after treatment and were seen on dates corresponding to the secretory phase of ovulatory cycles and concomitant to hormonal changes reflecting ovarian inhibition. In the coil group no abnormal EEG signs were observed in patients before during and after the regimen. 1 patient in the microdose group was found to have EEG abnormalities but the abnormalities were not attributed to the hormone content of the compound as abnormal EEG signs were found before and after treatment and were not concomitant to ovarian inhibition. No EEG abnormalities were observed in records taken during menstruation and during sleep of all the subjects.

Chlormadinone acetate 1-2-H3 (C.A.) ∗∗ or progesterone -7-H3 was administered intravenously to patients 8 hours before hysterectomy, and at this time tissue samples were taken for counting. The highest tissue concentration (dpm/g. wet tissue) of chlormadinone acetate (7 cases) in the tissues examined was found to be in the subcutaneous abdominal fat. All the patients were in different phases of the menstrual cycle. In the genital tract, the greatest amount of radioactivity was found in the fallopian tube, endocervix, or cervical mucus per se (6 out of 7 cases). The average urinary excretion of C.A. over 3 days was 17.5% of the total dose. In contrast, only 3 out of 6 patients treated with progesterone showed the highest concentration in the subcutaneous abdominal fat. These patients were in the proliferative phase of the cycle. The remaining patients, in the follicular phase, showed maximum concentration of radioactivity in skin, uterus, and uterine fibroids. In addition, progesterone was not concentrated by the mucous producing tissues of the genital tract in any of the patients. The average urinary excretion of progesterone over 3 days was 32.58% of the total dose.

Radioactively labelled d-Norgestrel * was injected intravenously to six volunteer patients 8 hrs prior to hysterectomy with the purpose of elucidating the site(s) of uptake and to correlate this with blood levels, urinary excretion and stage of the menstrual cycle. In all cases, a selective deposition of radioactive material was found in the endometrium regardless of the stage of their menstrual cycle. However, important quantitative differences were observed in the amount of radioactivity deposited at genital tract tissues and subcutaneous abdominal fat related to the stage of menstrual cycle. Radioactivity in blood during the first 8 hrs was composed mainly of free steroids; conjugated steroids composing the majority of the radioactivity found in blood by 120 hrs. Urinary excretion of radioactivity by 80 hrs was 43.38% of the administered dose during the estrogenic phase and 35.12% during the progestational phase. Deposition of radioactivity in the genital tract tissues may be related during the proliferative phase to estrogenic induction of steroid binding protein which may be a receptor. Contraceptive activity achieved by 30 μg of daily administration of d-Norgestrel may be related to selective endometrial concentration of this compound. This selective uptake was not seen previously for progesterone or chlormadinone acetate.*Kindly provided by Dr. H.J. Scharff (Schering/Berlin).

Human cornea were excised at the time of surgical procedure from patients undergoing radical surgery. Three separate incubations were performed in Eagle's minimum essential medium with [4- 14 C]-progesterone as precursor for five days. From the initial radioactivity, 6.9% was biotransformed to 6β-hydroxy-4-pregnene-3,20-dione. This steroid was identified by paper chromatography, derivative formation, and recrystallizations to constant S.A.

The zoapatle aqueous crude extract has been used in Mexico for the last 5 centuries for the induction of labor, treatment of post-partum bleeding problems, and as a menses inducer. Today, it is sold in street markets, and its long documented history of use by humans could be taken as indirect evidence of a lack of toxicity. Rigorous pharmacological and clinical studies described here, fully confirm the empirical observations.For the last 5 centuries in Mexico the zoapatle aqueous crude extract (ZACE) has been used for the induction of labor, treatment of postpartum bleeding problems, and as a menses inducer. Despite widespread and continuous use of ZACE on the part of the Mexican population and the absence of documented side effects or toxicological phenomenon associated with its use, it was decided to undertake acute and subacute toxicological studies prior to clinical studies in volunteer subjects in Mexico and Sweden. Plant specimens for the studies were collected in different batches at several locations near the National University of Mexico. The material was properly identified at the university's botanical herbarium. ZACE was prepared by boiling 100 g of dry leaves in 400 ml of distilled water for 20-30 minutes, filtered through gauze, and the final volume obtained by using a rotor evaporator 50-60 degrees Centigrade with light negative pressure, to 100 ml. The ZACE concentration was 1 g dry leaves/ml. The extract was tested in an "in vitro" guinea pig uterine strip assay. For the 1st series of studies in Mexico, 60 female Sprague-Dawley rats, 8 weeks old, and 6 mongrel female dogs, were used. 25 rats were used as control and 35 tested. All animals were maintained in a controlled laboratory condition with 12/12 hour light/darkness cycle. 2 dogs served as control, and 4 were treated. All experimental animals received, through a rubber cannula, 5 ml/KG of ZACE daily. Control animals received distilled water. Each animal was carefully observed for 30 minutes after fluid intake. At the end of the study, the rats were sacrificed with ether, and the dogs with phenobarbital. Complete anatomapathological examinations were performed. 6 control and 8 treated rats died during the study, and all deaths were associated with accidental placement of the rubber cannula. Behavioral changes were observed in neither rats nor dogs throughout the study. No changes were recorded in body weights. No macro nor microscopic alteration was found in either group of rats. Results of hematological examinations did not differ between the 2 groups. It was concluded that ZACE is devoid of acute and subacute toxicity. In addition to the clinical observations reported by Gallegos in Mexico, where 10 normally menstruating volunteer women received 15 or 30 gm/day of zoapatle dry leaves extract orally for 3-9 consecutive days without clinical evidence of any side effects, clinical investigations were conducted by Landgren et al. in Sweden. 6 women in early pregnancy were given zoapatle; 6 women in early pregnancy served as a control group. A significant dilation of the cervix was found in each of the subjects treated with zoapatle decocts. A menstrual like cramp was reported by all and bleeding occurred in 4 of 6 subjects. No gastrointestinal symptoms were observed. No pain or bleeding nor significant dilatation of the cervix was evident prior to surgery. Immediately prior to vacuum extraction, cervical dilatation was less than 5 mm in all subjects. In sum, ZACE was devoid of toxic complication or side effects.

A study was designed to establish the degrees by which additions of progesterone to long-term incubations of human endometria at various menstrual phases can modify enzymatic processes. Progesterone-67-tritiated was biotransformed by human proliferative and secretory endometria into: 6beta-hydroxy-4-pregnen-3- 20-dione 20alpha-hydroxy-4-pregnen-3-one and 20beta-hydroxy-4-pregnen-3-one 5alpha-pregnane-3-20-dione and 5beta-pregnane-3 20-dione. There was progesterone-67-tritiated biotransformation into the above metabolites in the absence (control) and in presence of 10 and 100 mcg/ml unlabeled progesterone. The biotransformation of the radioactive precursor was traced at incubation periods of 6 24 48 and 72 hours. At increasing dosages of labeled progesterone and longer incubation periods a decrease of total metabolites was observed. Linearity of the progesterone 67-tritiated biotransforming process is observed only in proliferative endometria controls. The proliferative endometria formed proportionately greater concentrations of highly polar compounds than the secretory one. Lesser amounts of pregnanediones were formed by the latter. There was no change in the C-20 reduced progesterone derivatives under any conditions. Further study is necessary to determine whether the in vitro endometrial capabilities are similar in tissue exposed to bioactive progestrone-releasing IUDs.(Authors modified)

A collection of Montanoa (Cerv) specimens was conducted throughout Mexico. Twenty-one specimens were classified, some of them grown in the greenhouse and transplanted in an agricultural experimental field station in the Valley of Mexico. In vitro uterotonic potency was assayed and the results expressed as equivalents of oxytocine, by using estrogenized guinea pig uterine strips. A great variation of uterotonic potency was observed among the wild plants. A clear decrease and uniformity of uterotonic potency was found in plants grown in the experimental field. The study points out the importance of ecological variations in expression of the plant's biological activity.21 specimens of the zoapatle Montanoa (Cerv) plant were collected from sites throughout Mexico with a variety of climatic, humidity, soil composition, and altitude conditions to assess their uterotonic properties. Some plants were then grown in an agricultural experimental field station of the National Institute of Agricultural Research, where all the variables were known, allowing for a valid comparison between species and plants. In vitro uterotonic potency was assayed and the results expressed as equivalents of oxytocin. The wild plants were found to vary greatly in uterotonic potency, perhaps reflecting differences in ecological conditions. The most active uterotonic zoapatle aqueous crude extract was the Montanoa frutescens, and the least active was M. mollissima, the former having approximately 4 times the uterotonic potency of the latter. The plants grown in the agricultural field station showed a considerable decrease in uterotonic potency, but valid comparisons must await assay in winter to match the stage in the plant life when the wild samples were assayed. Plant age, the part of the plant from which the uterotonic principle is extracted, climate, altitude, and soil constituents are other possible influences on uterotonic potency. A sample of the zoapatle aqueous crude extract with the highest uterotonic potency will be assayed for its luteolytic activity in an animal model.

In vitro histamine response in rat and guinea pig uterine strips was similar to the one observed with zoapatle aqueous crude extract (ZACE). The relaxing effect elicited by ZACE/ tomentosa on the in vitro rat uterine strips was not mediated by activation of H-2 histamine receptors; moreover, the increase on uterine contractility elicited by ZACE/tomentosa on the in vitro guinea pig uterine strip was not mediated through the activation of H-1 histamine receptors.

Post-partum bleeding was estimated during the first 72 hours in 106 women with normal delivery at term, all had live child with a body weight between 2.5 and 3.5 Kg. Three groups of women were studied: Group 1 comprised 41 volunteers with normal deliveries to whom no medication had been administered; Group 2 comprised 39 volunteers receiving 20 IU of oxytocin in 250 ml of a 5% glucose solution, immediately after delivery; and in Group 3, 26 volunteers received 0.2 mg of ergonovine maleate, orally three times daily for three days. The average post-partum blood loss in Group 1 during the first 72 hours after delivery was 151.5 ± 12.5 ml; for Group 2, 155.9 ± 13.9 ml; and for Group 3, 135.5 ± 15.9 ml. There were no significant differences among the groups and, most of the blood loss occurred during the first 24 hours after delivery.

16α-Hydroxy-ent-kauran-19-oic acid was isolated from Montanoa hibiscifolia . The effects of this acid and its methyl ester on the contractile activity of rat and guinea pig uterine horns were studied. Both inhibited spontaneous, oxytocin-induced and potassium-induced contractile activities. The inhibitory effect produced by the methyl ester was greater than that observed with the acid. The methyl ester was 2–5 times more potent than the acid upon spontaneous and potassium-induced contractions and 11–15 times more potent than the acid upon the contractile activity of uterine smooth muscle induced by oxytocin. Such effects were observed using bath concentrations of 6, 15, 30 and 60 μM of each compound.

Kauradienoic acid was obtained from the hexanic extract of M. tomentosa (Cerv) leaves by chromatographic separation. This substance influenced the in vitro contractility of the rat, dog and guinea pig uterine strips. It also induced strong contractions of the guinea pig uterus in vivo when administered intravenously, without changes on arterial blood pressure. The effects produced by the plant infusion, the hexanic extract and pure species were compared. The hexanic of other utero-active compounds in M. tomentosa in addition to those already described is discussed.The investigation of uterotonic effects of the hexanic extract (HE) of M. tomentosa leaves, as well as that (-)-kaura-9(11), 16-dien-19-oic acid, henceforth referred to as kauradienoic acid (KA) is described. Pregnant and nonpregnant female Wistar rats, guinea pigs, and dogs were used in this study. Strips obtained from 5 animals had spontaneous rhythmical contractions constant in amplitude and frequency. Muscular tone and frequency of contractions were notably increased by addition of 20 ul/ml of zoapatle aqueous crude extract (ZACE). The preparation returned to the original spontaneous contractility pattern after washing with a modified Ringer solution. Strips obtained from 5 gravid guinea pigs showed scarce spontaneous contractility which was strongly augmented by addition of ZACE. Regular spontaneous contractions were observed in over 30 assays of uterine strips obtained from 5 nongravid rats. The addition of ZACE drastically inhibited the spontaneous contractility. Constant and reproducible results were obtained in gravid rats by addition of ZACE. A gradual decrease in amplitude of contractions was observed. The addition of ZACE depressed the uterine contractility (UC) of 8 uterine strips from 2 animals. The addition of ZACE in gravid dogs in 8 uterine strips was accompanied by an increase of muscular tonus and frequency. A gradual decrease in the amplitude of the rat spontaneous UC was an effect produced by the HE. Addition of water suspension of HE elicited an immediate increase of guinea pig's uterine motility. Results were similar with those observed with ZACE. KA inhibited the "in vitro" rat UC and increased the "in vitro" guinea pig contractions of uterine musculature in a similar fashion as observed with ZACE and HE.

Oral administration of zoapatle aqueous crude extract (ZACE) made from Montanoa tomentosa or frutescens at doses of 5 mg/day from days 3 through 17 of pregnancy in rabbit, did not modify the normal plasma levels of progesterone. Pregnancy and delivery were similar in controls and zoapatle-treated animals.

Post-partum bleeding was measured for 72 hours in 66 volunteer healthy women after normal pregnancy and delivery at term. After complete placenta expulsion, a 220C Copper T was placed by the attending physician in 33 volunteer subjects before leaving the delivery room. Similar number of volunteer subjects served as a control. Post-partum bleeding during the 72 hours of study was 134.7 ± 16.3 ml for the IUD group and 155.9 ± 16.0 ml for the control group. No differences were detected among the two groups. Hemoglobin levels were the same for both groups, being at 72 hours after delivery, 11.9 ± 0.28 gm/ml for the IUD group and 12.2 ± 0.36 for controls. None of the women with immediate post-partum IUD insertion had any changes in the normal characteristics of their post-partum period.

A review of publications from 1529 to 1982 related to the zoapatle was made. Historical, biological, ethnobotanical, agricultural, clinical and chemical studies were noted; most of the early publications were written by Mexican scientists, in Spanish. More recent ones appear in the form of patents dealing with the isolation, characterization and total synthesis of some of its constituents. From this review, ideas emerged that might contribute to the understanding and rational use of this plant, or its chemical constituents, as a new, safe, reliable, inexpensive and - hopefully soon - a widely used interceptive agent.Zoapatle, the common name for Montanoa tomentosa, is a well-known plant in Mexico that has been used since the 16th century to induce labor or to increase both the tone and frequency of uterine contractions during labor. Since 1866, the plant has been recognized as a potential menses inducer. Pharmacological observations of zoapatle were made in Mexico in 1945 and focused on the administration of zoapatle aqueous crude extract (ZACE). Animal studies demonstrated increased uterine contractility. 9 compounds have been isolated from zoapatle. Much of the recent work with the material has been detailed in several patent applications. In 2 of 4 clinical studies on the use of ZACE, an increase in uterine contractility and cervical dilation among pregnant women was noted, and in another study, a cramping sensation was reported by nonpregnant women. The need to control for ecological variables which effect the plants seasonally was mentioned by 1 researcher who found working with the crude extract difficult and unpredictable. The lack of toxic effects was demonstrated in a study using animal models. Different effects were noted in a study which used zoapatanol obtained by total synthesis, including the lack of uterotonic response in rabbit tissue which warrants further investigation. The plant or its derivatives are potentially useful as interceptive agents but further research under controlled conditions is needed.

Human cornea excised from patients with wounded eyes were incubated in vitro for a 5-day period in the presence of [4-14C]-progesterone. The following C21 steroid metabolites were identified by paper chromatography, derivative formation, and crystallizations to specific activities: 20α-hydroxy-4-pregnen-3-one, 20β-hydroxy-4-pregnen-3-one, and 5α-pregnan-3,20-dione.

An estimate of the menstrual blood loss (MBL) in 29 women using the Cu-7 and in 33 women using the ML-250 IUDs was carried out. None of the women who took part in the study used hormonal contraceptives or IUDs for six months previous to the study. Before the insertion of the IUD, the MBL of each woman was measured for one or two cycles. Once the IUD was inserted, the MBL was measured at intervals of 1, 3, 6, 9 and 12 months after insertion. After this, the IUD was removed and the MBL measured each month thereafter for 3 months. In the pre-insertion period, the MBL was similar in both groups: 39.3 ± 5.9 for the group with the Cu-7, and 41.6 ± 5.5 ml for the one with the ML-250. After the insertion of the Cu-7, either a slight increase or no increase at all of MBL was observed during the 12 months of IUD use. When the IUD was removed, the menstrual blood loss level returned to normal, i.e. to the pre-insertion level.

Fifty-three volunteer women using Copper T 220C IUDs, complaining of increased menstrual bleeding, received per os 1 g, three times a day, of acetylsalicylic acid, for 5 days, during their menstrual periods. Menstrual bleeding for each patient was measured at least once before treatment. Bleeding estimates were also performed from the second to the fifth treatment cycle. From the 53 women admitted to the study, only 13 subjects (24.7%) had pre-treatment menstrual bleeding of more than 80 ml; 40 subjects had less than 80 ml. The group with hypermenorrhea had slightly decreased (not significant) the amount of menstrual blood loss with acetylsalicylic acid intake. On the other hand, 67.1% of women with bleeding less than 80 ml observed a significant increase in menstrual blood loss.

A review of new Zoapatle publications was made, including five chemical compounds characterized recently. Relative potency of Kaurenoic acid, Kauradienoic acid (its mixture), Zoapatanol and Montanol was estimated in an in vitro guinea pig uterine assay. Kaurene compounds were several times more potent than Montanol and Zoapatanol as uteroactive substances. Zoapatle aqueous crude extract made from Montanoa frutescens had a strong in vitro uterine potency and unique in vivo characteristics in several biological systems, described in detail in the following five publications.

The Effect Of Two Derivatives Of Kaurenoic Acid Isolated From Montanoa Frutescens Upon The Spontaneous Activity Of Estrogen-Dominated Rat and Guinea Pig Uteri Was Assessed. The Methyl Ester Of 15 A-Hydroxy-Dihydro Kaurenoic Acid Induced A Concentration-Related Decrease Of Spontaneous Activity Of Estrogen-Dominated Rat Uterus (Id 50 = 49.3 μ). This Effect Was Not Modified By The Presence Of Propranolol. The Methyl Ester Of 15-Keto-Dihydro Kaurenoic Acid Induced A Concentration-Dependent Increase Of Spontaneous Activity Of Estrogen-Dominated Rat Uterus (Ed 50 = 83.5 μm). This Increase Was Not Altered By The Presence Of Atropine Or Indomethacin. Both Compounds Induced An Increase Of Spontaneous Activity Of Estrogen-Dominated Guinea Pig Uterus In A Concentration-Related Manner. The Ed 50 Value For 15 α-Hydroxy Derivative Was 48.9 μm, Whereas The Ed 50 Value For 15 Keto Derivative Was 26.3 Yum, I.E. The Latter Was 1.16 Times More Potent Than The Former. These Increases Were Not Modified By Atropine Or ...