The direct assay of serum progesterone after denaturation of the binding proteins was investigated. 50ul of patients' serum was diluted with 750ul phosphate buffer (0.05M, pH 7.4) and heated to 65 degrees C for 20 minutes. After cooling, 300ul of the treated serum was reacted with a rabbit antiserum to progesterone-11 alpha-hemicuccinyl-bovine serum albumin conjugate (Bioclin, U.K) and 1,2,6,7, tritium labelled progesterone. Separation of bound and free fractions was achieved with dextran coated charcoal. The method correlated well (r = 0.98) with an established method involving ether extraction of progesterone prior to assay. The mean sensitivity was 2.01 nmol/L (range 1.90-2.23nmol/L). The proposed method considerably shortens assay time and removes a tedious and imprecise stage in the conventional method involving extraction of serum.
A double-antibody radioimmunoassay technique has been used to investigate the serum prolactin (hPRL) level in Malay females from premenarche to the postmenopause. The results showed that the hPRL level (mena ± SEM) in the premenarchal, postmenarchal and late pubertal/reproductive subjects were 23.6 ± 2.3, 19.1 ± 2.0 and 22.7 ± 1.9 ng/ml respectively. In premenopausal women, hPRL level (11.8 ± 2.4 ng/ml) was significantly reduced (p < 0.01) compared to that of late pubertal group; the level declined even further after menopause (9.5 ± 1.7 ng/ml). Although the difference in the mean prolactin levels between premenopause and postmenopause were not significant, 73% of the postmenopausal women had serum prolactin concentrations below 10 ng/ml compared to 44% of the premenopausal and 10% in late pubertal group.
A simple and sensitive double-antibody radioimmunoassay for human growth hormone (HGH) was developed, optimised and validated. The anti-hGH sera raised in 2 rabbits were highly specific with low cross-reactions of 0.19% and 0.3% with human placental lactogen and 0.21% and 0.13% with human prolactin. The mean sensitivity of the assay determined from 28 assays was found to be 0.4 +/- 0.2 mIU/L. Mean recovery of added exogenous hGH was 98.8 +/- 6.8%. Linearity studies of samples diluted at 1:2, 1:4 and 1:8 gave values of 101.3 +/- 5.3%, 109.6 +/- 13.4% and 97.3 +/- 13% respectively of those expected. The reproducibility of the assay was good; within assay coefficient of variation for serum samples with GH concentrations of 2.7, 13.6 and 28.2 mU/l ranged from 5.1 to 8.3% while the inter-assay precision varied from 4.9 to 10.3%. The in-house assay showed good correlation (r = 0.96, p less than 0.001) with a commercial HGH RIA kit (Dainabot, Japan). A reference normal adult fasting GH level of less than 7 mIU/l was established from 95 samples assayed by this method.
We assessed the analytical performance of the Abbott IMxTM immunoassay analyser for total beta human chorionic gonadotropin (Beta hCG), follicle-stimulating hormone (FSH) and luteinising hormone (LH). The within-run CVs for various analyte concentrations were 2% to 6% while those for between-run imprecision in routine assay ranged from 4% to 10%. IMxTM values correlated well with radioimmunoassay for beta hCG, and immunoradiometric assay for FSH and LH; the correlation coefficients (r) were 0.97, 0.99 and 0.98 for total beta hCG, FSH and LH respectively. The average sensitivities were approximately 3.1, 0.2 and 0.5 iu/l for beta hCG, FSH and LH, respectively. Sample carry-over was not detected and there was negligible cross-reaction between LH and beta hCG in the respective assays. The automatic sample dilution protocol for beta hCG was superior to the manual procedure. The IMxTM is easy to operate and is able to process 24 samples in 40-45 minutes.
Eighty-nine patients who had hydatidiform moles evacuated at the General Hospital, Kuala Lumpur, were followed with serum beta hCG determinations from October 1988 to June 1991. A regression curve for serum beta hCG, as measured by RIA, was derived from the results of 47 of the patients who demonstrated spontaneous regression of serum beta hCG titres. All 47 patients had normal serum titres at 135 days after evacuation. The mean time taken to reach normal level was 82.6 days, while the range was 39 to 135 days (5 to 19 weeks).
Serum human prolactin (hPRL) levels in Malay women during pregnancy, intrapartum and immediately postpartum have been invest£gated by means of a double-antibody radioimmunoassay technique. There was a progressioe rise of serum prolactin concentration from 31.9 ± 10.4 ng/ml in the first trimester to 242.0 ± 24.6 ng/ml at 36 weeks pregnancy unth. the mean values during the second and third trimester of 118.9 ± 12.7 ng/ml and 214.7 ± 10.3 ng/ml respectively. During intrapartum the concentration of hPRL was 191.9 ± 26.9 ng/ml and ithat of immediately postpartum was 178.3 ± 14.5 ng/ml. Suckling of the breast within 6 hours postpartum has resulted in a minor elevation of prolactin level, and this could have been due to the greater basal prolactin level, and/or the ineffectiveness of suckling as a potent stimulus during this period.
Sera from 80 Malaysians with confirmed hepatocellular carcinoma were tested for five markers of the hepatitis B virus, anti-HCV and anti-HDV by enzyme immunoassay, and alpha fetoprotein (AFP) was measured by radioimmunoassay. Of the patients, 98.8% had evidence of HBV infection and 75% were positive for HBsAg--which latter correlated with AFP raised above cut-off values of 500 ng/ml (P = 0.0001) and 200 ng/ml (P = 0.005). Males correlated significantly with the presence of HBsAg (P = 0.002). Thirty-one per cent of HBsAg positive patients were also positive for HBeAg and 74% for anti-HBe. Twenty per cent of the cases were concurrently positive for both HBsAg and anti-HBs. Six of 70 (8.6%) patients were positive for anti-HCV, of whom four were also positive for HBsAg. None of 67 patients tested for anti-HDV were positive. The results strongly indicate an important aetiological role for hepatitis B virus in causation of hepatocellular carcinoma among Malaysians.
Growth hormone (GH) levels were measured after a 75g oral glucose load (OGTT) in normal adults, patients with impaired glucose tolerance (IGT), insulin-dependent diabetes mellitus (IDDM) and acromegaly. Nadir GH levels at 2-hour post-OGTT in normal subjects ranged from 0.4 to 8.4 mIU/L, the 95% confidence interval being 0.4-4.4 mIU/L. In IGT and IDDM subjects basal fasting GH levels were not significantly different from normal and did not alter during OGTT. The high fasting GH level measured in one each of the IGT and IDDM patients was suppressible at 1-hour after glucose intake. In contrast, acromegalic patients had elevated fasting GH levels (11.8-178 mIU/L) although in 3 patients, the levels were mildly elevated and overlapped with normal. OGTT failed or only partially suppressed GH secretion in all acromegalics. Therefore, elevated fasting GH levels are not diagnostic and OGTT is required for accurate diagnosis and assessment of treatment of acromegalic patients.
62 cases of polycystic ovary syndrome (PCO) were reviewed with regards to their clinical and endocrine features. The subgroup of patients with acanthosis nigricans (AN) was further studied in detail. The prevalence of the syndrome was significantly higher in the Indian (35.5% of cases). Obesity, AN, hirsutism, non-insulin dependent diabetes mellitus (NIDDM) and raised level of serum testosterone were present in 77.1%, 74%, 79%, 21% and 48% of the cases respectively. Patients with AN was associated with higher body mass index, serum testosterone level, and prevalence of hirsutism and NIDDM than patients without AN. These observations are in keeping with the hypothesis that hyperinsulinemia may be of importance in the pathogenesis of a sub-group of PCO associated with insulin resistant states.
13 patients with the amenorrhoea-galactorrhoea syndrome who conceived during treatment with bromocriptine were reported. Mean period of amenorrhoea was 3.0 years. In ten patients galactorrhoea was noted for a mean period of 4.2 years while in three it was discovered during examination. Seven patients presented with primary infertility. Menses returned in all cases after a mean duration of 2 months of treatment with bromocriptine at an average dose of 5. 86 mg daily. Mean serum prolactin was 4344 mUll (range 750 mU/l to 23,000 mU/l) before treatment and this declined to 186 mU/l with treatment. Seven patients became pregnant 5 to 25 months of treatment while six conceived after first menses. 21 pregnancies resulted from the thirteen patients. There was one spontaneous abortion and one premature delivery in which the baby died. Of the 16 live- births, there were twelve girls and four boys and their mean birth-weight was 2932 g. All were normal at birth and during subsequent developments except one with congenital dislocation of hip. It is concluded that bromocriptine is effective in restoring menstrual cycles and fertility by lowering serum prolactin in patients with the amenorrhoea-galactorrhoea syndrome. Bromocriptine may be safe for use during pregnancy, but it is suggested that the medication should be stopped immediately after conception unless tumour growth is apparent.
To compare the effects of dietary palmitic acid (16:0) vs oleic acid (18:1) on serum lipids, lipoproteins, and plasma eicosanoids, 33 normocholesterolemic subjects (20 males, 13 females; ages 22-41 years) were challenged with a coconut oil-rich diet for 4 weeks. Subsequently they were assigned to either a palm olein-rich or olive oil-rich diet followed by a dietary crossover during two consecutive 6-week periods. Each test oil served as the sole cooking oil and contributed 23% of dietary energy or two-thirds of the total daily fat intake. Dietary myristic acid (14:0) and lauric acid (12:0) from coconut oil significantly raised all the serum lipid and lipoprotein parameters measured. Subsequent one-to-one exchange of 7% energy between 16:0 (palm olein diet) and 18:1 (olive oil diet) resulted in identical serum total cholesterol (192, 193 mg/dl), low-density lipoprotein cholesterol (LDL-C) (130, 131 mg/dl), high-density lipoprotein cholesterol (HDL-C) (41, 42 mg/dl), and triglyceride (TG) (108, 106 mg/dl) concentrations. Effects attributed to gender included higher HDL in females and higher TG in males associated with the tendency for higher LDL and LDL/HDL ratios in men. However, both sexes were equally responsive to changes in dietary fat saturation. The results indicate that in healthy, normocholesterolemic humans, dietary 16:0 can be exchanged for 18:1 within the range of these fatty acids normally present in typical diets without affecting the serum lipoprotein cholesterol concentration or distribution. In addition, replacement of 12:0 + 14:0 by 16:0 + 18:1, but especially 16:0 or some component of palm olein, appeared to have a beneficial impact on an important index of thrombogenesis, i.e., the thromboxane/prostacyclin ratio in plasma.
BACKGROUND: Iron deficiency and its consequent anemia constitute the commonest micronutrient deficiency in the world.
OBJECTIVE: We investigated whether long-term, weekly iron-folate supplements administered at school would improve hemoglobin and ferritin concentrations in adolescent girls, including those with mild-to-moderate anemia and hemoglobin concentrations indicating borderline anemia.
DESIGN: Subjects were 266 girls with hemoglobin concentrations of 80-119.9 g/L (group A) and 358 girls with hemoglobin concentrations of 120-130 g/L (group B) who were otherwise healthy. Two hundred sixty-six girls in group A and 268 girls in group B were randomly assigned to receive either 60 or 120 mg Fe plus 3.5 mg folic acid weekly for 22 wk. Ninety of the girls in group B were randomly assigned to receive only 5 mg folic acid weekly. Capillary hemoglobin and plasma ferritin were measured at baseline and after 12 and 22 wk of supplementation.
RESULTS: By the end of the study, 2% of the girls had dropped out and > 96% had taken > or = 20 of the 22 tablets; side effects were minimal. Mean plasma ferritin increased significantly in all iron-supplemented groups, independently of initial hemoglobin values and iron doses. Ferritin concentrations decreased in the girls supplemented with folic acid only. As expected, hemoglobin responses to iron were higher in group A than in group B and increases were positively correlated with initial plasma ferritin. Hemoglobin failed to respond to folate supplementation if initial plasma ferritin concentrations were low. Mean hemoglobin increased significantly and consistently in relation to the length of treatment.
CONCLUSION: Long-term, weekly iron-folate supplementation was found to be a practical, safe, effective, and inexpensive method for improving iron nutrition in adolescent schoolgirls.
Comment in: Picciano MF. Iron and folate supplementation: an effective intervention in
adolescent females. Am J Clin Nutr. 1999 Jun;69(6):1069-70. PubMed PMID:
10357724. https://academic.oup.com/ajcn/article/69/6/1069/4714856
Study site: 3 secondary schools, Samarahan district, Sarawak, Malaysia