The vitamin B6 requirement in oral contraceptive users. II. Assessment by tryptophan metabolites, vitamin B6, and pyridoxic acid levels in urine.

The requirement for vitamin B6 in oral contraceptive users was studied in 8 college-age women who used combined (7) or sequential (1) oral contraceptives. The subjects and 8 controls consumed a basal diet supplemented to result in daily intake of 2.06 mg pyridoxine hydrochloride for 10 days (predepletion) and then containing only .36 mg of vitamin B6 for 32 days. After the depletion period, the diet was supplemented with pyridoxine hydrochloride to increase the intake of B6 to .96, 1.56, and 5.06 mg for 8, 9, and 7 days respectively. Complete 14-hour urine collections were analyzed for xanthurenic acid, kynurenic acids, kynurenine, and 3-hydroxykynurenine after administration of a l load-dose of 2 gm L-trytophan on days 2, 11, 18, 25, 32, 39, 43, 50 , 59, and 66 for the subjects and days 2 and 10 for the controls. Pretryptophan urine was analyzed for vitamin B6. Posttryptophan urine was analyzed for 4-pyridoxic acid. It was found that during the depletion phase the excretion of tryptophan metabolites increased significantly. Excretion dropped significantly upon supplementation with 1.56 or 5.06 mg of vitamin B6, returning values to normal. Levels of vitamin B6 and 4-pyridoxic acid in the urine decreased during depletion to be restored to normal upon supplementation with 1.56 mg/day. Since an intake of 5.0 mg vitamin B6 caused a loss of the vitamin in the urine and all levels were returned to normal with an intake of 1.56 mg, it is suggested that 1.5 mg of vitamin B6 is sufficient to meet the needs of most oral contraceptive users and that there is no significant difference in the vitamin B6 requirement of oral contraceptive users and nonusers.     

Assessment of vitamin B 6 status. Studies on pregnant women and oral contraceptive users.

The vitamin B6 status of 10 pregnant women (third trimester), 9 oral contraceptive agent users, and 12 notnpregnant women (controls) was investigated over a 10-week period by means of the erythrocyte glutamate-oxaloacetate transaminase (E-GOT) activation test and by measurements of whole blood pyridoxal phosphate levels. Blood pyridoxal phosphate levels in oral contraceptive agent users (7.6 plus or minus 1.1 ng/ml) were significantly lower than in the control group (9.6 plus or minus 1.7 mg/ml)indicating a relative vitamin B6 deficiency. Pyridoxal phosphate levels in pregnant women(5.1 plus or minus 1.3ng/ml) were even lower; in fact, no overlap was found in individual mean values between the pregnant and control groups. The E-GOT activation test did not indicate a vitamin deficiency in the pregnant women or in oral contraceptive agent users and the E-GOT activation factor did not correlate with blood pyridoxal phosphate levels unless results obtained after pyridoxine hydrochloride (20 mg/day) administration were included. The E-GOT activation test appears to be a poor indicator of vitamin B6 status, except in pronounced deficiency as it is less responsive to vitamin depletion than blood pyridoxal phosphate levels, and suffers from relatively large variations in individual control values. This may be a result of factors unrelated to vitamin B6 as blood pyridoxal phosphate levels remained fairly constant in the individuals investigated.     

Humoral immunity in oral contraceptive users. I. Plasma immunoglobulin levels

The worldwide acceptance of steroid-based oral contraception makes it imperative that the effect of these agents on the immune system is understood. Nevertheless, information regarding the effect of steroid-based oral contraception on plasma immunoglobulin (Ig) levels is often conflicting. In this report immunoglobulin levels in the plasma of females taking steroid-based oral contraceptives are measured in a novel manner using an enzyme-linked immunosorbent assay (ELISA). No significant alterations in the levels of IgG or IgM are reported.     

Immune studies in oral contraceptive users.

Some of the parameters of immune status-percentage of B and T lymphocyte subpopulation, PHA-induced lymphocyte transformation measured by 3H-thymidine incorporation (PILT) and levels of total haemolytic complement (CH5O) were studied in Indian women from low income group who were using estrogen-progestogen combination pills. There were no differences in percentage of B and T lymphocyte subpopulation or PILT between OC users and the control group. However, CH5O levels were significantly lower in OC users. The depression in circulating complement levels in OC users is intriguing and suggests the possibility that complement system and circulating immune complexes may be altered in OC users.     

Multivitamin supplementation in oral contraceptive users.

The effects of oral contraceptives (OC) containing 30 micrograms of ethinyl oestradiol and of subsequent multivitamin and folic acid supplementation on vitamin A, total B2 [including its three individual constituents, i.e. riboflavine, RB; flavine-mono-nucleotide, FMN; and flavine-adenine-dinucleotide, FAD], B12, C and folate concentration in serum and red blood cells have been studied in a group of 59 non-pregnant female volunteers. The group taking OC comprised 28 women while 31 women were included in the group of non-OC users serving as the controls. The women were studied for four cycles. Blood samples were taken on days 3 and 23 of the first cycle to obtain baseline values of each analyte. Multivitamin and folic acid supplementation started on day 1 of the second cycle and this was continued daily throughout three consecutive cycles until the end of the study. Vitamin A levels were significantly higher and vitamin B12 levels were significantly lower in the group using OC. Comparison of the baseline values of vitamin total B2, FAD, C, serum and red blood cell folate as determined on days 3 and 23 of the first cycle of the two groups compared revealed no significant differences. Multivitamin and folic acid supplementation did not affect the concentrations of vitamin A and vitamin B12 with either group, whereas all other vitamins increased significantly in both groups. The consistency of each effect of multivitamin supplementation between the two groups was also tested. The degree of these effects was not statistically different between both groups. The results suggest that the vitamin status is indeed affected by OC treatment, but the effects of multivitamin supplementation are not different in OC and non-OC users. Supplementation during OC use or just after discontinuing treatment cannot be justified for healthy young women. However, in the case of women with a critical vitamin balance or higher folate needs, multivitamin supplementation may be considered.     

Fibrinolytic activity of uterine fluid in oral contraceptive users

The fibrinolytic activity of uterine fluid increased gradually during the menstrual cycle in oral contraceptive (OC) users. This is in contrast to the low activity found during the luteal phase in non-users. It is concluded that the “synthetic gestagens” contained in the OCs used in this study influence the fibrinolytic activity of uterine fluid in a different way than does native progesterone during the normal luteal phase. This may imply an additional contraceptive effect of OCs.     

Effect of the administration of vitamin B6 at two levels of intake on xanthurenic acid excretion among oral contraceptive pill users

Researchers compared data on 30 25-35 year old women who took the combined oral contraceptive (OC) Ovlar (0.05 mg ethinyl estradiol and 0.5 mg dinorgestrel) for 2-5 years with data on 10 women who did not take OCs to determine the longterm effect of Ovlar on vitamin B6 metabolism and urinary Xanthurenic acid (XA). Cases received either 50 mg or 100 mg of vitamin B6 tablets every day for 4 weeks. Mean XA excretion/24 hours stood much higher among OC users than among the controls (12.51 vs. 2.33; p.01), e.g., XA excretion among OC users ranged from 3.45 to 34.95 mcg/24 hours. Among the OC users, mean urinary XA excretion/24 hours was significantly higher before 50 mg vitamin B6 administration than it was after its administration (13.5 mcg vs. 2.71 mcg; p.01). Administration of 100 mg vitamin B6 had the same effect (11.52 mcg vs. 2.26 mcg; p.01). XA excretion was basically the same for the 2 OC user groups (50 mg and 100 mg administration of vitamin B6). Both doses of vitamin B6 brought about comparable XA excretion levels as the control group. The researchers concluded that 50 mg of vitamin B6 taken daily for 4 weeks is sufficient to correct vitamin B6 deficiency and changed metabolism among OC users. Research has demonstrated that OCs alter vitamin B6 tryptophan metabolism in 2 ways. They increase hepatic tryptophan oxygenase activity resulting in a surplus of tryptophan further in the pathway to niacin. This effect explains the higher production of several metabolites. OCs also reduce the proclivity for kynureninase, the pyridoxal phosphate of vitamin B6 dependent enzyme of tryptophan metabolism perhaps increasing the need for plasma pyridoxal phosphate.     

The psychical and sexual adaptation of oral contraceptive users

Oral contraceptives have effects on the sexual and emotional life of women. They are successful in: 1) reducing menstrual pain and premenstrual tension, 2) maintaining contraceptive effectiveness, and 3) reducing depression or neurasthenic problems among 10 to 20% of women. Negative effects of oral contraceptives generally stem from conscious or unconscious sexual or psychical conflicts. Negative motivating factors (to use or not to use oral contraceptives) include pressure from spouse, mass media publicity, using mechanical protection to discourage frequency of intercourse, and confused feelings of sterility or feminine identity.     

Serotonin metabolism and depression in oral contraceptive users

Serotonin and nicotinic acid ribonucleotide metabolic pathways of tryptophan metabolism were studied before and after tryptophan load test in thirty women using oral contraceptive steroids for a period of 2 to 5 years. Ten of them were suffering from depression. Another ten healthy women participated in this study as a control group. Twenty-four-hour urinary excretion of serotonin, 5-hydroxy indole acetic acid and total 5-hydroxyindoles were estimated as indices of serotonin pathway metabolites, while xanthrenate excretion was determined as an index of tryptophan oxygenase pathway.Plasma cortisol, urinary 17-oxosteroids and 17-hydroxycorticosteroids were determined to assess adrenal cortical function. Urinary creatinine output was assayed to check the adequacy of 24-hr urine collection. The changes induced by oral contraceptive steroids on tryptophan and corticosteroid metabolism were correlated with the associated depression. Changes in serotonin metabolism were demonstrated in the depression group before and after tryptophan load test, while in the non-depression group, these changes were only demonstrated after tryptophan load test.Results indicated that alterations in tryptophan metabolism are usually well compensated in the non-depression group but may accentuate or precipitate the development of depression in susceptible women.     

The classical and alternate pathways of complement in oral contraceptive users

Whole complement (CH50), C3, C4, the alternate pathway activity of complement (APH50) and factor B were measured in 159 women currently taking oral contraceptives and in 186 women who were not taking the pill. The mean levels of all components of the complement, except APH50, were found to be elevated in current users compared to non-users. The elevation in serum levels was seen in the first year after initiation of oral contraceptive use. Thereafter, levels changed little with duration of use. Among women who stopped using oral contraceptives, complement levels were similar to those of women who had never used the pill. Results appeared to be similar irrespective of the type of progestagen included in the oral contraceptive. These data provide support for the view that oral contraceptives have no adverse effect on the classical and the alternate pathways of complement.     

Riboflavin nutritional status and absorption in oral contraceptive users and nonusers.

Riboflavin nutritional status, based on erythrocyte glutathione reductase (EGR) determinations, and the absorption of the vitamin after 10-mg oral doses were assessed in a group of oral contraceptive users (group OC, n = 18) and in a group of nonusers (group NOC, n = 17). Before administration of test doses of riboflavin, mean EGR levels of 4.06 (group OC) and 4.44 (group NOC) mumoles glutathione reduced per minute per gram of hemoglobin were not significantly different. After oral administration of riboflavin, a significant increase in EGR levels was observed within each group although no significant differences were detected between groups. Mean EGR levels continued to be slightly lower for group OC compared to group NOC. Oral absorption of riboflavin based on urinary excretion results was not significantly different between the groups. These findings indicate OC use does not significantly affect riboflavin nutritional status in a subject population having adequate nutrition of moderate to high socioeconomic status. Significant reductions in EGR levels which have been previously reported for OC users appear to be associated with a low socioeconomic status.     

The level of dietary protein affects the vitamin B-6 requirement of cats

A depletion-repletion model was used to study the effect of the level of dietary protein on the vitamin B-6 requirement of growing kittens. Twenty kittens were given a pyridoxine-free diet for 42 d to deplete vitamin B-6 reserves. They then were divided into four groups of five kittens each. Two groups were fed diets containing 30% casein with either 1.0 (Group 30-1) or 2.0 (Group 30-2) mg pyridoxine/kg diet and two groups were fed 60% casein diets with either 1.0 (Group 60-1) or 2.0 (Group 60-2) mg pyridoxine/kg diet for 44 d. During repletion, body weight gain of kittens from Group 30-2 was higher than that of kittens from the other groups. Body weight gains of kittens from Groups 30-1 and 60-2 were higher than that of kittens from Group 60-1, but there was no difference in body weight gain between Groups 30-1 and 60-2. At 44 d of repletion, kittens from Group 60-1 generally had higher plasma concentrations of free tyrosine and cystathionine, lower plasma B-6 vitamin concentrations and a higher urinary oxalate excretion than did kittens from the other groups. These findings indicate that the vitamin B-6 requirement of growing kittens--as is true of humans, chickens and mice--is positively related to the level of protein in the diet. For a 30% casein diet, the vitamin B-6 requirement was greater than or equal to 1.0 but less than 2.0 mg pyridoxine/kg diet; for a 60% casein diet, the requirement was greater than or equal to 2.0 mg pyridoxine/kg diet.     

Increased capillary permeability for plasma proteins in oral contraceptive users.

The transcapillary fluid balance was examined in eleven women before administration of a monophasic oral contraceptive (desogestrel 0.15 mg, ethinylestradiol 0.03 mg), and after three and six months of use. The interstitial colloid osmotic pressure was measured by the "wick" method, and the interstitial hydrostatic pressure by the "wick-in-needle" method in subcutaneous tissue on thorax and leg. During the six-month observation period, the following changes were observed: Plasma colloid osmotic pressure decreased (mean 1.8 mmHg, p = 0.047), as well as serum albumin (mean 5.1 g/l, p = 0.0006), total protein concentration (mean 2.8 g/l, p = 0.0006), hemoglobin (mean 0.5 g/dl, p = 0.014) and hematocrit (mean 1.8%, p = 0.047). Blood pressure and body weight remained unchanged, but foot volume showed a significant increase. The colloid osmotic pressure gradient (plasma-interstitium) was significantly reduced. The results indicate an increase in plasma volume in addition to an increased capillary permeability to plasma proteins during oral contraceptive use. We suggest that the observed changes in transcapillary fluid balance is caused by the estrogen component of the oral contraceptive pill.     

Antimullerian hormone and obesity: insights in oral contraceptive users

Background

The study was conducted to examine the impact of oral contraceptives (OCs) on serum antimullerian hormone (AMH) levels by obesity status in reproductive-age women.

Study Design

Ovulatory women, ages 18-35 years, of normal (<25 kg/m²; n=10) and obese (>30 kg/m²; n=10) body mass index (BMI) received a low-dose OC (20 mcg ethinyl estradiol/100 mcg levonorgestrel) for two cycles. Serum samples obtained at several time points during active pill use and hormone-free intervals were analyzed for AMH, follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol and inhibin B.

Results

AMH levels did not differ by OC cycle day in either BMI group. On average, AMH levels were 34% lower in the obese group (2.9±2.1 vs. 4.4±1.8 ng/mL, p<.05). Modeling to determine differences in AMH throughout the cycle based on obesity status demonstrated significantly lower levels (p<.05), whereas serum AMH, FSH, LH, estradiol and inhibin B levels revealed no correlations when all time points were included.

Conclusions

In reproductive-age women, serum AMH levels do not appear to fluctuate during OC use, but AMH levels are significantly lower in obese women. Lower levels do not appear to be due to differences in gonadotropin levels or ovarian activity.     

Humoral immunity in oral contraceptive users. II. In vitro immunoglobulin production

The worldwide acceptance of steroid-based oral contraception makes it imperative that the effect of these agents on the immune system is understood. Nevertheless, information regarding the effect of steroid-based oral contraception on humoral immunoregulation is limited. In this report the in vitro production of IgG and IgM is measured following stimulation with either the T-dependent activator pokeweed mitogen (PWM) or the T-independent activator fixed/killed Staphylococcus aureus Cowan I (StaCw). No significant differences are observed between the in vitro IgG or IgM levels following stimulation with PWM or StaCw for females taking steroid-based oral contraceptives and females not taking steroid-based oral contraceptives. We conclude that humoral immunoregulation is unaltered in steroid-based oral contraceptive users.