Calcium (hydrogen-1-malate) hexahydrate on Echeveria gibbiflora leaves and its effect on sperm cells.

Echeveria gibbiflora is a plant widely used for its contraceptive activity in traditional Mexican medicine. Data on calcium crystals in plants are not outstanding. In the case of the Echeveria gibbiflora leaves, however, its quality, quantity, and salt type are quite surprising; one striking result of its X-ray crystallographic data shows the presence of calcium bis (hydrogen-1-malate) hexahydrate [2(C4H5O(5)1), Ca(1)2+, 6(H2O1)]. This highly soluble compound might explain the rapid shape changes of calcium crystals. Because SEM-EDS analysis shows that calcium malate crystals were obtained in a highly pure state and the immobilization and agglutination pattern that OBACE show on human and bull spermatozoa are not found even when high concentrations of calcium bis (hydrogen-1-malate) hexahydrate salt are present it is not feasible to involucrate molecules as calcium malate as part of the OBACE contraceptive activity.

Male contraception, IV: hypotonic-like effect from Echeveria gibbiflora on human sperm.

At present there is no accepted method for the regulation of male fertility. The most appropriate form of contraception would undoubtedly be to develop from traditional plant-derived folk drugs a contraceptive male method. Hypoosmotic shock is used for validate plasma membrane function and its fertilizing capacity of human sperm. Such effect is induced in human sperm in the presence of a purified fraction from Echeveria gibbiflora (PFEG) aqueous crude extract. The hypotonic-like effect included a distension of the plasma membrane over the acrosome region and in some occasions around the sperm middle piece. An enhanced activity of the immobilizing and agglutination effects was induced instantaneously after the addition of PEFG versus aqueous crude extract activity (OBACE). Using electron microscopy it was possible to observe a deposit of a "sticky" dense material intercalated along the plasma membrane. The membrane was sealed making it impossible to measure the viability or metabolic activity of the treated sperm by the fluorescence (FDA-IP) technique. It was also possible to corroborate the dispersion of the acrosomal content and the disappearance of the acrosomal and nuclear membrane. Results that makes PFEG hypotonic-like effect a serious candidate to conduct a study to determine the predicting capacity of this compound in human infertility and suggest that the plant may yield a compound suitable for use as male contraceptive agent.

Effects of a purified fraction from Echeveria gibbiflora aqueous crude extract on guinea-pig spermatozoa.

Guinea-pig spermatozoa in the presence of a purified fraction from Echeveria gibbiflora aqueous crude extract suffer a hypotonic-like effect. The phenomena exhibited included a distension of the plasma membrane over the acrosome region, inducing the formation of a huge 'head-bubble'. The agglutination effect was so enhanced that instead of inducing sperm clusters, it produced cane-like 'stalk' structures. The immobilizing activity was induced instantaneously after the addition of the purified fraction. At electron microscope level it was possible to observe a heavy amount of electron dense material of the purified fraction embedded or intercalated along the plasma membrane. It was also possible to corroborate the dispersion of the acrosomal content and the disappearance of the external acrosome membrane. The purified fraction induced loosening of the plasma membrane all along the sperm cell, however, the distension of the membrane was only produced in the apical portion of the sperm head and not in the post equatorial region. The results suggest that the plant may yield a compound suitable for use as a vaginal barrier or male contraceptive agent.

Immobilizing / agglutination effects of crude extract of crassulaceae plants on human sperm: screening study.

PIP: 17 species from 6 genera of the Crassulaceae family of Mexican plants were tested for sperm agglutination, killing, and immobilizing activity. These plants, which grow in the valley of Mexico and surrounding areas, have long been used as a vaginal postcoital douche. The genera tested were Aeonium, Crassula, Echeveria, Kalanchoe, Pachyphytum, and Sedum. The plants were harvested in the flowering phase, which varied over the season for species. Aqueous crude extracts were prepared by grinding 260 gm fresh leaves, filtering through gauze, centrifuging at 3000 rpm for 15 minutes, lyophilizing, and dialyzing against 2 changes of deionized water. The final extracts were reconstituted 1 mg/ml in Keyhani and Storey medium. Motility, viability, and agglutination of washed human sperm were estimated. All 17 species exhibited immobilizing, agglutinating, and sperm killing activity. Aqueous extracts and infusions of several other plants from 5 other families were also tested without effect on sperm. Preliminary work suggests that the active agent is a micromolecule rather than a macromolecular protein.^ieng

RNA metabolism during the sexual differentiation of rat hypothalamus.

Modifications in the basal molecular biology parameters (total concentrations of RNA, DNA, proteins, rRNA, tRNa, free and polysomal bound poly-A+ mRNA) have been determined daily in the growing hypothalamus of male and female rats from day 1 to day 8 after birth. Changes observed in the parameters studied in this work occurred mainly in the first 48 h after birth. In males tRNA and free mRNA (f-mRNA) contents decreased from day 1 to day 2 and then their concentrations remained more or less constant. Total mRNA significantly decreased from day 1 to day 3 and showed a further significant decrease from day 6 to day 8. Polysomal bound-mRNA (b-mRNA) decreased from day 1 to day 3, then increased to day 6, and finally decreased once more from day 6 to day 8. The b-mRNA/f-mRNA, mRNA/rRNA and mRNA/total RNA ratios showed a bimodal behavior with a first peak on day 2, and a second, smaller peak, on days 6-7. The changes observed in the females on the first 2-3 days of life were the inverse of those observed in the males, most of the parameters studied showed a sharp increase from day 1 to day 2 or to day 3 and then a drastic decrease. The only exception to this behavior was the b-mRNA/f-mRNA ratio which showed a small decrease from day 1 to day 2, followed by a continuous increase from day 2 to day 8. b-mRNA concentrations, after the sharp decrease from day 2 to day 3 of life, increased from day 3 to day 7.(ABSTRACT TRUNCATED AT 250 WORDS)

[Agglutinating and immobilizing activity of an ethanol extract of Bursera fagaroides on human and other mammalian spermatozoa]. / Actividad aglutinante e inmovilizante del extracto etanólico de Bursera fagaroides sobre espermatozoides de humano y de otros mamíferos.

Human spermatozoa and those obtained from mouse epididymis incubated in ethanolic from mouse epididymis incubated in ethanolic extracts of the cortex of Bursera fagaroides became agglutinated and immobilized in 100% with 30% of viability decrease. This double effect occurred in 50% of pig'ssperm and 50% of rabbit sperm when the concentration was increased tenfold. The pattern of progressive motility was affected 80% of rat's spermatocytes without agglutination. The values remained the same when the extracts were partially purified with salts of Zn++ and Ba++ or by filtration in a sephadex g-10 column. Analysis with thin layer chromatography of both purified extracts produced 3 compounds in comparison with the ethanolic unpurified extract. The extract of the plant Bursera fagaroides contains 3 compounds, which apparently are glycosides with a potent activity upon agglutination -immobilization and a low effect upon spermatocytes viability, which might be used as contraceptives .