Comparison of some trace elements concentration in blood, tumor free breast and tumor tissues of women with benign and malignant breast lesions: an Indian study.

Fifty women residing in and around New Delhi, India and identified to have benign (25 nos.) and malignant (25 nos.) breast lesions were studied for the first time to access the association between environmental exposure to lead and risk of breast cancer and to determine the potential of changes in trace elements concentration as a diagnostic marker and/or its etiological involvement in the disease. Blood, tumor tissue and breast adipose tissue from tumor free area from each patient of the two groups, collected at the time of lumpectomy or mastectomy (only blood sample was collected from disease free control group), were analyzed to determine the concentration of Pb, Zn, Cu, Fe and Ca using Atomic Absorption Spectrometry. Blood lead was significantly higher in malignant cases than in those of benign and control (p<0.05 each). Lead level was also higher in tumor tissue when compared with their respective normal tumor free breast tissue, though non-significant, in both benign and malignant cases. Interestingly, Zn, Fe, and Ca levels were higher in blood of malignant cases than in those of their benign counterparts. Furthermore, these metals were also higher in tumor of malignant and benign cases as compared to normal tumor free breast tissue, many of them statistically significant (p<0.05/0.01/0.001). However, Cu level was insignificantly lower in the blood and tumor tissue of malignant cases when compared with their benign counterparts while it was significantly higher (p<0.05) in tumor of benign cases when compared with those of their respective normal tumor free breast tissue. There were statistically significant correlations between lead and trace element levels only in normal tumor free breast tissue of benign and malignant cases (r=0.41-0.73; p<0.05-0.001) but neither in blood nor tumor tissue of the two groups. These results suggest that in the backdrop of existing experimental and epidemiological evidences exposure to lead may be one of the risk factors for breast lesions; though it warrants further investigation. Further, modulation of trace elements level in both benign and malignant breast diseases patients may be of potential to be used as diagnostic marker of the disease process and its possible relationship etiologically.

Isolation and culture of hamster ectoplacental cone trophoblasts: an in vitro study on the cell types and their growth pattern.

The method of isolation of trophoblast cell types from ectoplacental cone of hamster embryo of day 8 post coitum (plug day as day 1) and examination of their growth pattern in vitro is presented in this study. Based on their size, three types of trophoblast cells were identified. (1) the smaller cells having clear cytoplasm formed the major portion (70% to 80%) of the total cell population (2) moderately bigger cells having mono or binucleated appearance and containing minute granules in the nuclear region formed 5% to 10% and (3) extra large and multinucleated cells shared 10% to 20% of the total cell number. While the smaller and slightly bigger cells showed moderate growth the larger ones had extensive growth and were seen to acquire different shapes on extending the duration of culture, such as fusiform, dumb-bell, polygonal, rectangular or flowery. The extremities of the cytoplasmic growth of these cells were seen to be thickened at one end giving the impression of a pad-like structure. The significance of these adaptations are not known at present.

Syntheses and bioevaluation of substituted dihydropyridines for pregnancy-interceptive activity in hamsters.

A number of 2,6-dimethyl-3,5-bis(methoxycarbonyl)-4-substituted-1,4- dihydropyridines were synthesized and evaluated for pregnancy-interceptive activity in mated hamsters. Out of 24 compounds, 12, 15, 21, 22, 28, and 34 caused a marked reduction in the number of implantations when administered on days 3-8 postcoitum. In an in vitro competition assay, none of the compounds exhibited noticeable binding affinity for uterine progesterone receptors. The results reported here have helped to identify new leads for developing pregnancy-interceptive agents and the active compounds do not seem to elicit their interceptive effect through receptor-mediated inhibition of progesterone action in hamster uterus.

Non-steroidal menses-regulating agents: the present status.

PIP: Contraception researchers are just beginning to study the likelihood of developing menses regulators, most likely in the form of a once-a-month pill. Some advantages of this pill would be self-administration and short-term use without long-term side effects. Both pregnant and nonpregnant women could benefit from its use. Such a pill would either prevent implantation or cause the embryo to detach from the endometrium during the early stages of implantation. Researchers at the Central Drug Institute in Lucknow, India, have reviewed the sequence of events in the endometrium and blastocyst during the implantation and pre-implantation stages and have examined changes which may be applicable to menses induction. Likely biochemical target sites are prevention of endometrial preparations for blastocyte attachment, modification of the physicochemical environment of the uterus to thwart implantation, and interference with the functions of the utero-placental junction for facilitating detachment/resorption of the implanted blastocyst. In vivo studies in laboratory animals and in vitro studies are using various steroidal and nonsteroidal compounds. Mifepristone (RU-486) has already demonstrated effective contragestational activity. Research needs for successful development of menses inducers include: identification of biochemical markers which facilitate in vitro screening of pregnancy interceptive activity; a guarantee of a correlation of in vitro and in vivo contragestational efficacy of test compounds; identification of the pharmacophores which would steer the test compounds to the uterus in concentrations sufficient to intercept pregnancy; and search for exact biochemical target site(s) affected by compounds demonstrating a contragestational effect in laboratory animals.^ieng

Monte Carlo computer simulation of the aqueous hydration of the glycine zwitterion at 25 degree C.

Monte Carlo computer simulation on a dilute aqueous solution of the glycine zwitterion are reported. The results are presented in terms of the Quasi-Component Distribution Functions (QCDF) of Ben Naim and partitioned into atomic and functional group contributions using the Proximity Criterion. The Proximity Criterion analysis has been extended to orientational properties and a new normalization procedure has been introduced for the radial distribution functions obtained by the Proximity Criterion. The solvation environment of the glycine zwitterion is found to contain, on the average, 14.4 water molecules out of which 3.2 belong to the ammonium group, 6.1 to the methylene group and 5.1 to the carboxyl group. The importance of the many-body statistical mechanical approach to hydration is emphasized by our finding that the configuration corresponding to the absolute minimum of the glycine zwitterion-water potential surface was found to have negligible statistical weight in the aqueous simulation.

Surface morphology of hamster (Mesocricetus auratus) decidual cells in vitro.

Cell surface morphology of hamster decidual cells isolated from day 8 implantation swellings was studied, using both phase-contrast and scanning electron microscopy. Two kinds of cells, fibroblastic and epithelioid, were identified in cultures examined by phase-contrast microscopy. Fibroblastic cells were spindle-shaped, having pointed or blunt terminals on one end and bifid or webbed projections at the other end. Epithelioid cells, on the other hand, were flat and discoid, having a distinctively ruffled plasma membrane. Further, the plasma membrane of epithelioid cells formed rope-like or flange-like processes. The significance of such adaptations is discussed.

Phenotypical response in different cell types of rat uterus to centchroman. A qualitative analysis.

To assess the alleged estrogenic and antiestrogenic profile of centchroman, a phenotypical response was studied in native uterine cells or migrant blood cells in adult ovariectomized rats under the influence of estradiol-dipropionate (EDP) and centchroman (CC) given per se and together. Histologic observations were divided arbitrarily into the following regions: luminal epithelium, superficial and deep stroma, and the two layers of muscularis. Glandular cell proliferation, mitoses, and vascularity were examined additionally. It was found that CC per se stimulated luminal epithelial cell height and glandular cell proliferation better than EDP. Leukocytic infiltration and vascularity were also more pronounced in the uteri of CC-treated rats. Furthermore, except for a slight reduction in glandular cell size and edematous condition of circular muscle layer, the changes remained unaffected after conjoint treatment of CC and EDP. The observations revealed that some of the nongenomic responses, elicited by the rat uterus after CC treatment, seem to be unrelated to the hormonal attributes of the molecule.

Effect of inhibitors of enzymes involved in polyamine biosynthesis pathway on pregnancy in mouse and hamster.

The syntheses of polyamines, which play a definitive role in cell proliferation and tissue growth during early embryogenesis, are believed to be principally regulated by two enzymes: ornithine decarboxylase (ODC) and polyamine oxidase (PAO). However, when the level of enzyme ODC goes down, another enzyme known as S-adenosyl methyl decarboxylase (SAMDC) is believed to up-regulate the biosynthetic pathway. In the present study, the effects of inhibitors of enzymes SAMDC and PAO were determined in mated hamster and mouse in peri-and immediate postimplantation stages of pregnancy. It was noticed that, whereas the SAMDC inhibitor (MDL-73815) failed to arrest the pregnancy in the hamster in spite of raising the dose to 100 mg/kg, the PAO inhibitor (MDL-72527DA) was found to be 100% effective at 75 mg/kg in both species. The results confirmed the role of enzyme PAO in promoting early development of the embryo.

Pregnane derivatives as pregnancy interceptive agents: efficacy determination on growing trophoblasts (in vitro) and in pregnant hamsters (in vivo).

An in vitro test system was standardized to study potentiality of five hormonally inert pregnane derivatives on growing trophoblasts isolated from ectoplacental cone (EPC) of day 8 hamster embryo. Cells were incubated with different concentrations of respective compounds in surface droplets. The response was determined by analyzing the sequence of changes in cell morphology like attachment, growth, proliferation, differentiation and/or degeneration within 24 or 48 h following seeding. The in vivo efficacy of these compounds was determined in hamster during peri- and immediate post-implantation periods (days 3-8 post coitum). Two compounds 88/583 and 88/585 were found to inhibit not only growth and proliferation of the cells but caused total degeneration within 24 h. The same compounds induced partial to complete resorption of the foetuses in treated animals. Whereas, the other three compounds 88/506, 88/594 and 89/43 that showed lack of comparable potentiality in vitro were found to be equally ineffective in vivo. The results indicate a positive correlationship between in vitro and in vivo activity.

Search for new chemical entities as menses inducing agents.

In continuation of an ongoing program on developing nonsteroidal pregnancy interceptives to be used as a menses regulating agent, a new class of compounds belonging to 3-substituted amino-1-aryl-6-hydroxy-hex-2-ene-1-ones series has been investigated for pregnancy interceptive activity in the hamster and rat. The compounds were administered (subcutaneous) on days 4-8 (hamster) and 5-9 (rat) post coitum (PC). The animals were laparotomized on days 12 (hamster) and 16 (rat) PC. To derive percent efficacy, the total number of implantation was divided by the number of normal and resorbed implantations. Among the 14 compounds evaluated, three were found to intercept pregnancy by 100%. Another compound was active by 75%, whereas the rest were inactive. None of the active compounds were, however, active in rat with this schedule. Results indicate that the observed species- and schedule-specific activity owes its origins to differences in the implantation physiology and early post-implantation development between the two species. The study, nevertheless, offers an insight to the new class of compounds for this activity.

Pregnancy interceptive efficacy and biological profile of 3-amino-6,7-dimethoxy-1H-pyrazolo [3,4-b] quinoline (compound 85/83) in rodents.

Administration of compound 85/83 during the peri- and post-implantation period intercepted pregnancy in hamster and guinea pig by parenteral route and in hamster by oral route also. The m.e.d. for hamster and guinea pig was 10 and 20 mg/kg, respectively; lower doses were less effective. Restricting the administration to early post-implantation schedule interrupted pregnancy partially in both species. The compound was, however, ineffective in rat and in the pre-implantation schedule (days 1-4 post-coitum) in hamster. When tested in vitro on growing trophoblasts at 13.8 x 10(-5) M concentration, it prevented growth and caused degeneration of the cells within 24 h; lower concentration (9.2 x 10(-5) M) was less effective. The compound was found to be devoid of estrogenic, antiestrogenic, progestational and antiprogestational properties in conventional bioassays. In hormone competition assays, its relative binding affinity (RBA) to estrogen receptor was negligible (0.002% of estradiol-17 beta), while for uterine cytosol progesterone receptors in rabbit and hamster was 0.06 and 0.08% of progesterone, respectively. The compound 85/83 appears to intercept pregnancy by interfering with development of trophoblast cells.

Emetine ditartrate: a possible lead for emergency contraception.

Interception of pregnancy in its initial stage is an attractive and viable approach to contraception. A chemical agent, taken within the first few days of missed menses, intercepts the conception, which is expelled with menstrual flow. The main targets of such agents are the uterus, blastocyst and the growing trophoblasts, whose nutritional requirement is inhibited. Our previous work has identified several nonsteroidal chemical entities as pregnancy interceptives in rodents and infrahuman primates. However, none reached clinical stage due to their ineffectiveness by oral route. Nevertheless, parallel to these rationally designed synthetic compounds, a program was ongoing to identify natural product(s) that can be used as interceptives. We are reporting for the first time the detailed profile of emetine ditartrate, a compound whose pregnancy interceptive efficacy has been studied in mouse, rat, hamster, guinea pig and rabbit by oral and intravaginal routes of administration. By the oral route, the compound caused 100% resorption of the fetuses in rat, hamster and guinea pig at 6.0, 5.0 and 3.0 mg/kg, respectively, on administration during peri- and early postimplantation periods of pregnancy (depending upon the day of implantation in each species). By intravaginal route, the compound was administered once in the form of a vaginal pessary on the day of implantation in respective species; interception of pregnancy was not achieved completely in rat and hamster at doses four to five times the oral dose in multi-day schedule. However, in guinea pig and rabbit it was fully effective at 7.0 and 70.0 mg/animal, respectively. The compound was devoid of estrogenic, antiestrogenic and progestational activity but possessed mild antiprogestational activity at the high dose in vivo. In in vitro assay, however, it did not show any significant binding to estrogen and progesterone receptors. The mode of action of the compound was found to be mainly on the uterus and early embryos around implantation, possibly on the trophoblasts and endometrial cells at the attachment site. The absence of 100% efficacy in rat and hamster by intravaginal route, but not by oral route, is possibly due to poor absorption of the compound through the vagina in these species. The guinea pig and rabbit, therefore, seem the better species for evaluating the efficacy of the compound administered by the vaginal route.

Contraceptive and hormonal properties of a new 1,4-dihydro-2-oxoquinoline derivative (compound 84-182) in rodents and rhesus monkeys.

Compound 84-182 prevented pregnancy when administered subcutaneously at 10 mg/kg dose on days 3-8 post-coitum in hamsters and on days 6-10 post-coitum in guinea pigs. At lower doses, while in hamsters there was a marked reduction in implantation number, majority of implantations in guinea pigs showed signs of resorption. The compound was ineffective when administered at 10 mg/kg dose on days 1-3 or 6-7 post-coitum in hamsters and on days 1-5 or 4-8 post-coitum in rats. In rhesus monkeys, treatment with the compound at 5 and 10 mg/kg doses on days 16-21 of the menstrual cycle induced frank vaginal bleeding between days 21 and 24. Treatment on days 21-30 or after confirmation of pregnancy on days 32-36 was ineffective. In conventional bioassays, the compound was devoid of any estrogenic, antiestrogenic, progestational, antiprogestational, androgenic or antiandrogenic properties at the contraceptive dose. In competitive protein binding assay, the compound showed relative binding affinity (RBA) of less than 0.1% and 0.28% of progesterone, respectively, for rabbit and hamster uterine cytosol progesterone receptors. Its RBA for rat uterine cytosol estrogen receptors was less than 0.1% of estradiol-17 beta.

Ultrastructure of cellular components of human trophoblasts during early pregnancy.

Two cell types, the cyto- and syncytio-trophoblasts, were identified in human chorionic villi of 6-10 weeks' gestation. The intracellular organization of these cells was examined. Ultrathin sections of small pieces of chorionic villi revealed the presence of a multinucleate syncytiotrophoblastic layer, whose surface was covered with microvilli. The cytotrophoblasts, however, had a single large nucleus with a prominent nucleolus. An interesting feature of the basement membrane of these cells was the presence of aggregates of dark granules in samples of the earlier gestational age (6-8 weeks) and granular bodies having a dense outer ring and a translucent inner ring with a lucid central area in samples of 8-10 weeks' gestation. Both types of granules are mineralized and are assumed to perform a buffering role for maintaining the neutrality of the layer.

Blood lead and zinc in pregnant women and their offspring in intrauterine growth retardation cases.

As part of our program to investigate the possible role of environmental pollutants in the incidence of intrauterine-growth retardation (IUGR) in India, we determined the lead and zinc levels in mothers and neonatal blood, collected at parturition, in cases with normal and IUGR babies. Both maternal and cord blood lead levels were significantly higher in IUGR cases than in normal cases (p < 0.05). The mean level of zinc was also higher in maternal blood of IUGR cases. Significantly, the mean cord blood lead level was > 10 microg/dL, which is greater than Centers for Disease Control's intervention level, in 54% of newborns. A good correlation (r = 0.53, p < 0.01) between maternal and cord blood lead levels confirmed the transfer of lead from mother to the fetus. There was a weak but significant relationhsip between cord blood lead levels and birth weight of newborns (r = -0.22, p < 0.05). The study may serve as a pointer to the perils of in utero exposure to chemical contaminants and a call for measures by the public health authorities for a continuous bio-monitoring program to evaluate impact of environmental pollutants on women and children's health.

Some essential elements in maternal and cord blood in relation to birth weight and gestational age of the baby.

Maternal and cord blood were collected from 54 Indian women at parturition and analyzed for Zn, Cu, and Fe by flame atomic absorption spectrophotometry to determine the relationship between levels of these elements in mother's and infant's blood and maternal age, birth weight, and gestational age of the baby. The blood Zn level of mothers in the age group 24-28 yr was significantly higher than those of mothers in the age group of 18-23 yr (p<0.05). Similarly, mothers in the 24 to 28-yr group also had higher blood Fe level than mothers in the group 29-38 yr (p<0.05). The levels of Zn, Cu, and Fe were higher in the maternal blood and lower, but not significantly, in the cord blood of low-birth-weight babies than in those of normal-birth-weight babies. However, differences in the levels of Zn, Cu, and Fe between maternal and cord blood of the two birth-weight groups was statistically significant. There were no significant differences in the levels of the three elements in maternal or cord blood by the gestational age of the baby. A weak but significant correlation was found between the birth weight of the baby and the Fe level in the cord blood (r=0.26; p<0.05). Also, weak significant correlations were observed between gestational age of the baby and Fe (r=0.23; p<0.05) and Cu (r=0.31; p<0.05) levels in the cord blood. Although, there are many confounders of low birth weight and preterm deliveries, a diminished placental transfer of these essential elements could be one of the several etiological factors for low birth weight of newborns.

Environmental exposure to lead as a risk for prostate cancer.

OBJECTIVE: To evaluate the possible role of environmental exposure to lead as a risk factor for prostate pathology in patients suffering from prostate cancer (PCA) and benign prostate hyperplasia (BPH). METHODS: Blood lead (BPb) level was determined in PCA and BPH cases using a graphite furnace Atomic Absorption Spectrophotometer and compared with those in a control group living in the similar socioeconomic environment. RESULTS: BPb was significantly higher in PCA and BPH cases than in normals (P < 0.05). Blood levels of zinc and copper were significantly lower in PCA and BPH cases when compared with controls (P < 0.05). In all the three groups, a statistically significant positive correlation between lead and thiobarbituric acid reactive substances (TBARS) measured as malondialdehyde, and negative correlation between blood lead and antioxidant GSH level, indicative of possible generation of reactive oxygen species, were also observed after adjusting for age as a possible confounders. However, positive association between blood lead and TBARS was relatively higher in PCA patients (r = 0.77, P < 0.05) than in BPH (r = 0.32, P < 0.05) and normal (r = 0.30, P < 0.05). CONCLUSION: These results with limited power seem to suggest for the first time that environmental exposure of aging males to lead may be a risk factor for prostate cancer and/or benign prostate hyperplasia possibly through generation of reactive oxygen species and/or reducing the level of zinc which acts as a cellular growth protector.

Ultrastructure of the yolk substance in preimplantation embryos of the gerbil.

The yolk substance or deutoplasm in preimplantation embryos of the Mongolian Gerbil was observed to be composed of tubule-like structures which were grouped in slightly wavy bundles running in various directions and occupying the major cytoplasmic space in the cell. The tubules were about 70 nm wide and, where cut longitudinally, had a maximal length of 2 micrometer in the micrographs. This deutoplasm structure has no similarity to that observed in other species, thus supporting the earlier assumption that the ultrastructure of deutoplasm is species specific.

Decidual cell reaction: relative response to traumatization in uterus of rat, hamster and guinea pig.

Relative decidual cell response to traumatization was studied in the uterus of immature rat, hamster and guinea pig. A single dose of progesterone (1.0 mg/animal) caused 285 +/- 41 and 459 +/- 58% uterotrophy in traumatized horn of rat and hamster respectively, while in guinea pig increasing doses of progesterone (1, 2 and 5 mg/animal), several combinations of estrogen and progesterone or alteration in treatment schedule of the two hormones as well as extending the day of traumatization failed to elicit comparable uterotrophic response. Uterus of immature hamster was found to be most responsive to traumatization and that of guinea pig the least.

Easy criterion for detection of mating in guinea pig.

Existing criteria for determining receptivity to mating in female guinea pig has been redefined on the basis of kind of cells present in vaginal smear following its opening. The animals that showed not only cornified but also predominantly cornified cells over the leucocytes show higher sperm positivity rate. It is inferred that continuous cohabitation of these animals invariably leads to successful mating.