Effect of oral contraceptive used by technician on sensitivity of progesterone radioimmunoassay.

Gonadotropin and steroid radioimmunoassays (RIAs) have been run in our laboratory for the past 6 years. Recently, the progesterone RIA was unpredictable when performed by one technician. After checking the mechanics and reagents, it was discovered that the technician had changed from Ortho Novum 1 + 50 to Ovral about the time the RIA became erratic. Standard curves of progesterone RIAs performed while the technician was taking either Ortho Novum 1 + 50 or Ovral, or on no contraceptive are reported. The data suggest that a probably exhaled metabolic product of norgestrel, the synthetic progestin in Ovral, interferes with the progesterone RIA and indicate an additional variable investigators must eliminate in performing RIAs.     

The characterization of sulphated metabolites of norethindrone in human milk after oral administration of contraceptive steroids

The excretion of ethynyl steroids in milk from a lactating woman taking a daily dose of an oral contraceptive (Conlumin) containing 1 mg of norethindrone and 50 micrograms of mestranol has been studied. Milk was diluted with aq. triethylamine sulphate and steroids were extracted on a Sep-Pak C18 cartridge at 60-64 degrees C. Groups of unconjugated steroids, glucuronides, mono- and disulphates were separated on triethylaminohydroxypropyl Sephadex LH-20. Following hydrolysis and further purification, steroids possessing an ethynyl-substituent were isolated by chromatography on sulphohydroxypropyl Sephadex LH-20 in silver form. Gas chromatographic-mass spectrometric analysis of the O-methyloxime-trimethylsilyl ether derivatives of these steroids, showed the presence of norethindrone and mestranol in the free fraction and of tetrahydro metabolites of norethindrone with 3 alpha,5 alpha, 3 alpha,5 beta and 3 beta,5 alpha configurations in the mono- and disulphate fractions. The disulphate of the 3 alpha,5 alpha isomer was the most abundant ethynyl steroid in milk after 13 days of administration. The site of conjugation of the monosulphates was established by acetylation prior to solvolysis and analysis by gas chromatography-mass spectrometry. This showed that the 3 alpha,5 alpha isomer was conjugated mainly in the 17 beta-position while the 3 alpha,5 beta isomer was conjugated at C-3.     

Genotoxic effect of Anovlar 21, an oral contraceptive, on mouse bone marrow

Anovlar 21, a combination drug containing the oestrogen ethinyloestradiol and the progestin norethisterone acetate, was studied for its in vivo genotoxic effect on the bone marrow cells of Swiss albino mice. The chromosomal aberration assay and the micronucleus test were employed for the study. 0.08, 0.4, 0.8, 1.6, 3.2, 4.8, 6.4 and 8.0 mg/kg/day of the drug was orally administered for 15 consecutive days to mice. Bone marrow preparations were made 24 h after the final feeding. The lowest dose, 0.08 mg/kg, represents the human therapeutic range. Marrow preparations of mice fed 0.8 mg/kg/day for 15 days were made at 6, 12, 24, 48 and 96 h, and 1, 2 and 3 weeks and a time-yield analysis was carried out. Statistically significant increases in chromosomal aberrations were observed in animal groups fed doses of greater than or equal to 0.4 mg/kg/day. In the time-response study, the maximum frequency of aberrations was noted at 24 h, thereafter decreasing gradually with increasing time. But the drug did not induce a significant increase in the number of micronuclei in bone marrow erythrocytes at any of the doses or time intervals studied.     

Effect of fluoride on cariogenic oral microorganisms (an in vitro study)

The effect of sodium fluoride and sodium monofluorophosphate at concentrations of 1, 5, 10, 50, 100 and 1000 mg/l in phosphate buffer (pH 6.5) as well as in UHT milk were studied on cultures and suspensions of Streptococcus mutans, Lactobacillus acidophilus and Candida albicans. Using serial tenfold dilutions up to 10(-7) of 24-48 hour cultures, a subsequent 0, 60 and 120 min incubation caused no decrease in the number of CFUs. Growth kinetic studies in the Bioscreen biophotometer (Labsystem, Finland) revealed that sodium fluoride in different concentrations (from 0.875 mg/l up to 500 mg/l) influenced the growth dynamics of S. mutans and C. albicans: the exponential phase flattened out at the highest fluoride concentrations (500 mg/l) present in the growth media. The lag phase of C. albicans became longer. The results of these experiments indicate that sodium fluoride administered at higher concentrations than the usual caries preventive dosage made the generation time of cariogenic oral bacteria and fungi longer, slowing down their multiplication.     

LM and SEM study on the swordfish (Xiphias gladius) tongue

Swordfish (Xiphias gladius L. 1758) is a predatory and migratory fish. Its characteristic feature is a flat and sharp upper jaw forming a “sword”. The adaptation of vertebrates, including fish, to their environment is strictly related to the capacity of feeding and is carried out by often severe modifications of the anatomy of the buccal cavity, especially of the tongue. The aim of this study is, using light and scanning electron microscopy and considering that no data are so far available about the morphology of the tongue in this species, to analyse the anatomical characteristics of the tongue, especially its dorsal surface. The tongue shows a triangular shape and an apex, a body and a root. By SEM the presence of several small denticles and filiform papillae on the latero-ventral body was demonstrated while no taste buds or other sensitive structures are observed. LM shows a squamous stratified epithelium, becoming simple cuboidal around the denticles. Therefore this study could add further data to the knowledges of the fish oral cavity morphology supporting the hypothesis that the modifications and evolution of the tongue anatomy are, also in fish, related to the environment and especially to the feeding habits.     

SEM study on cytotoxic effect of monocrotophos (MCP) on lungs of rat

Monocrotophos (MCP) on oral administration (0.28 mg/100 g of body wt. i.e. 1/5th of LD50) to female rats for 15 and 30 days damaged alveolar walls lined by type II cells (great alveolar cells); clara cells (non-ciliated cells) lining bronchiolar epithelium; and emphysematous lesions due to loss of inter-alveolar walls. This led to increase in surface tension in lung due to decrease in secretion of surfactant as a result of necrosis of great alveolar cells and clara cells resulting in hypoxia. This effect was time dependent. In R group (15 days without pesticide after 30 days daily oral treatment), the toxic effects mentioned above still persisted which revealed non-repair of necrosis caused by MCP.     

Effect of exercise and oral contraceptive agents on fibrinolytic potential in trained females

It has been shown that physical exercise increases blood fibrinolytic potential, primarily by inducing a release of extrinsic plasminogen activator from the vessel wall. Synthetic estrogens have also been reported to influence fibrinolytic activity. The effect of exercise and the possible additional effect of oral contraceptive agents (OCA) on the fibronolytic system were studied in 20 competitive female rowers. Ten females used OCA (users), and 10 others did not (nonusers). All participants were subjected to standardized exhaustive exercise. Preexercise data revealed higher factor XII, total plasminogen, and free plasminogen levels together with a significantly lower C1-inactivator level in the group of users. No differences were observed in prekallikrein, high-molecular-weight kininogen, alpha 2-antiplasmin, alpha 2-macroglobulin, antithrombin III, and histidine-rich glycoprotein plasma levels. The factor XII-dependent fibrinolytic activator activity and the extrinsic (tissue-type) plasminogen activator were significantly higher; however, the urokinase-like fibrinolytic activator activity was significantly lower. These observations suggest a greater susceptibility to activation of the fibrinolytic pathways during OCA medication. Exercise resulted in a decrease of all factors under study but an increase in all fibrinolytic activities. No differences were observed between the two groups in the percentages of change that occurred with exercise.     

Irreversible protein binding of norethisterone (norethindrone) epoxide.

14,15-3H-Norethisterone-4 beta, 5 beta-epoxide, a metabolite of norethisterone, was incubated with several proteins and nucleic acids. After 30 min incubation 0.19 nmol of the epoxide were irreversibly bound per mg albumin which contains free sulfhydryl groups; proteins without SH-groups, such as concanavalin A, gamma-globulin, DNA and RNA, did not irreversibly bind norethisterone epoxide. A superoxide (O2) generating enzyme system comprised of xanthine oxidase and hypoxanthine was capable of catalyzing the irreversible binding of the parent compound, norethisterone, to albumin, indicating that an oxidation product was formed which reacted with the protein. When norethisterone epoxide was incubated for 60 min with hepatic microsomes of rats in absence of NADPH, about 2.0 nmol of the epoxide were irreversibly incorporated per mg microsomal protein. This binding was increased to 5.2 nmol by addition of a NADPH regenerating system. Addition of glutathione and cytosol decreased only the NADPH-dependent protein binding; phenobarbital pretreatment of rats induced this NADPH-dependent binding of norethisterone epoxide to microsomal protein by a factor of 2. In presence of NADPH, binding of the epoxide to microsomal protein depended on substrate concentration used. The results indicate that norethisterone epoxide is able to chemically react with proteins. In addition, hepatic microsomal enzymes convert the epoxide to another metabolite which also can react with proteins.     

Effect of a steroidal oral contraceptive on intestinal absorptive functions in proteins deficient rat

The effects of steroidal oral contraceptive norethynodrel plus ethinylestradiol-3-methyl ether (SOC) at a daily dose of 5 mg: 0.06 mg per kg body weight for 28 days on intestinal absorptive functions have been investigated in protein-deficient female albino rats. The administration of this contraceptive caused significant increase in glucose and amino acids uptake but had no effect on calcium and zinc uptake in pair-fed as well as in protein-deficient rat. Further studies carried out on glucose transport system showed that the transport of sodium-dependent glucose was significantly enhanced while that of sodium-independent glucose remained unaltered in drug-treated animals. Kinetic studies of glucose transport in the presence of sodium ions revealed that SOC treatment affected the rate of uptake of glucose by elevating Vmax, but the apparent Kt value remained the same in treated and untreated animals.