The relationship of fetuin-A and lactoferrin with bone mass in elderly women.

SUMMARY: The relationships of fetuin-A and lactoferrin to bone-related phenotypes were investigated in elderly women. Fetuin-A was associated not only with bone mineral density (BMD) but also with bone resorption marker suggesting an influence of fetuin-A on osteoclasts. INTRODUCTION: The aim of this study is to investigate the relationship of bone-related phenotypes in elderly women with circulating fetuin-A and lactoferrin. METHODS: Eighty-two elderly women were studied. Serum fetuin-A, lactoferrin, C-terminal telopeptide of type I collagen (CTx), total procollagen type 1 amino-terminal propeptide, and plasma intact parathyroid hormone (PTH) were analyzed. BMD of the lumbar spine at L2-4 and at the femoral neck was measured. RESULTS: Serum fetuin-A was significantly associated with L2-4 BMD (r = 0.23, P < 0.05). After controlling for age and body weight, the association remained statistically significant. There was a significant association between serum fetuin-A and serum CTx (r = -0.37, P < 0.001). The association between fetuin-A and L2-4 BMD no longer existed after controlling for serum CTx. There were positive associations of circulating lactoferrin with plasma PTH (r = 0.24, P < 0.05) and serum CTx (r = 0.26, P < 0.05). No association between serum lactoferrin and BMD at the lumbar spine or femoral neck was detected. CONCLUSIONS: Circulating fetuin-A is related to bone mass and bone resorption markers in elderly women. Lactoferrin, in contrast, is associated only with bone resorption markers.

Chronic metabolic acidosis alters osteoblast differentiation from human mesenchymal stem cells.

Bone histology of distal renal tubular acidosis patients showed decreased bone formation with impaired bone matrix mineralization that is not entirely explained by an alteration in the mineral balance. Data from in vitro studies suggests a direct inhibitory effect of metabolic acidosis on osteoblast function. We investigated the effects of chronic metabolic acidosis on osteoblast differentiation from mesenchymal stem cells (MSCs). Human MSCs were allowed to differentiate into osteoblasts in culture. Concentrated hydrochloric acid was added to the medium to lower the bicarbonate concentration and pH. The expression of various osteoblastic genes and proteins and bone matrix mineralization were examined. Chronic metabolic acidosis enhanced the messenger RNA (mRNA) and protein expression of early osteoblast transcription factor, runx-2, whereas inhibiting osterix and having no effect on ATF-4. The expression of type I collagen, the most abundant bone matrix protein, was increased following the same pattern of runx-2. Likewise, metabolic acidosis slightly enhanced the expression of mature osteoblastic gene, osteocalcin. Study on mineralization revealed suppressed alkaline phosphatase mRNA and enzyme activity. Despite the augmented collagen deposit in acidic culture, bone matrix mineralization was impaired. In conclusion, chronic metabolic acidosis alters osteoblast differentiation from MSCs through its diverse effect on osteoblastic genes and proteins resulting in an impairment of bone formation.

A specific haplotype in the 3' end of estrogen-receptor alpha gene is associated with low bone mineral density in premenopausal women and increased risk of postmenopausal osteoporosis.

It is well established that the development of postmenopausal osteoporosis is under genetic influence. We have recently identified a synonymous single nucleotide polymorphism (SNP) in exon 8 of estrogen receptor-alpha (ERalpha) gene in the vicinity of the stop codon (G2014A) that is associated with an increased risk of postmenopausal osteoporosis. In the present study, we attempted to locate SNPs in the 3'-unstranslated region (3'UTR) of the ERalpha gene that are in linkage disequilibrium with the exon 8 SNP and assessed their utilization in the risk assessment of postmenopausal osteoporosis in 352 Thai postmenopausal women. The association with bone mineral density (BMD) in premenopausal women was also investigated in 202 premenopausal women. A C to A SNP 1,748 nucleotides distal to the end of the stop codon (C3768A) was identified. The C3768A SNP was not overrepresented in subjects with osteoporosis. However, the presence of the A-C haplotype allele based on the A2014 and C3768 alleles was significantly related to the risk of osteoporosis independently of age, body weight, the G2014A and C3768A SNPs (odds ratio 2.36, 95% CI 1.42-3.91). Moreover, the presence of the A-C haplotype allele was associated with lower femoral neck BMD in premenopausal women ( P =0.05). We concluded that a specific haplotype in the 3' end of the ERalpha gene is associated with lower BMD in premenopausal women and is associated with a higher risk of osteoporosis in postmenopausal women. It is likely that the haplotype allele exerts its influence on bone as early as during young adulthood to increase the risk of osteoporosis later in life.

Differential associations of residual estradiol levels with bone mineral density and serum lipids in postmenopausal women with osteoporosis.

OBJECTIVES: To examine the associations of residual endogenous estradiol (E2) to bone mineral density (BMD) and lipid concentrations in elderly women. METHODS: Subjects consisted of 59 elderly postmenopausal women with vertebral or femoral osteoporosis. BMD was measured at L2-4 and femoral neck by dual-energy X-ray absorptiometry (DEXA). Residual E2 concentrations were assessed by a sensitive radioimmunoassay. Data were expressed as mean +/- S.E.M. RESULTS: The age of the subjects was 65.2 +/- 0.8 years with 18.9 +/- 1.0 years postmenopausal. The mean residual E2 concentration was 6.0 +/- 0.5 pg/ml. There was a correlation between E2 levels and BMD at L2-4 (r = 0.32, P < 0.01) while no association was found at the femoral neck. The association between E2 and L2-4 BMD persisted after adjusting for years since menopause and body weight (r = 0.33, P < 0.05). With regard to serum lipid concentrations, no association of serum total cholesterol, LDL-cholesterol, HDL-cholesterol or triglyceride concentrations with residual E2 was found. CONCLUSIONS: Our findings confirm the role of residual endogenous E2 in the determination of bone mass in postmenopausal women with osteoporosis. The effect of residual E2 appears to be skeletal specific and possess no association with serum lipid concentrations.

Differences in bone mineral density and lifestyle factors of postmenopausal women living in Bangkok and other provinces.

UNLABELLED: Decreased bone mineral density (BMD) with age is an increasing health problem, especially for postmenopausal women. Multiple factors have been reported to affect BMD including both genetic and environmental factors such as calcium intake and physical activity. For Thailand, people residing in different regions may differ in BMD due to these factors. However, there is a paucity of data concerning this issue. The objectives of this study were to identify the lifestyle factors which may influence BMD and to investigate the association between BMD and these factors in postmenopausal women who have been living in Bangkok and other provinces in Thailand. Subjects consisted of 466 postmenopausal women aged 46-90 years including 236 Bangkokians (116 early postmenopausals and 120 late postmenopausals) and 230 non-Bangkokians (134 early postmenopausals and 96 late postmenopausals). All were healthy and ambulatory. BMD was measured by dual energy X-ray absorptiometry (DEXA, Expert XL). Calcium intake was assessed by food-frequency questionnaire. Data were expressed by mean + /- SEM. There were 22 per cent (n=52), 5.9 per cent (n=14), and 4.2 per cent (n=10) of postmenopausal Bangkokians while 13.9 per cent (n=32), 4.3 per cent (n=10), and 2.2 per cent (n=5) of postmenopausal non-Bangkokians who had low BMD at spine, femoral neck, and at both sites, respectively. Spine BMD (SPBMD) and femoral neck BMD (FNBMD) increased significantly across the quartiles of calcium intake in both groups of subjects (P<0.05) and a significant difference was found between the lowest and the highest quartiles of calcium intake (P<0.05). Moreover, BMD at both regions was shown to be correlated with calcium intake, exercise and sunlight exposure in these subjects (P<0.001). Further analysis revealed higher BMD at spine (0.992 + 0.02 vs 0.945 +/- 0.02 g/cm2, P<0.05) and at femur (0.780 +/- 0.01 vs 0.740 +/- 0.01 g/cm2, P<0.05), calcium intake (348.9 +/- 12.7 vs 316.3 +/- 8.0 mg/day, P<0.05), exercise (2.8 +/- 0.1 vs 2.4 +/- 0.1 h/wk, P<0.001) and sunlight exposure (2.9 +/- 0.06 vs 1.9 +/- 0.04 h/day, P<0.001) were found in late postmenopausal women in other provinces than their counterparts in Bangkok. Nevertheless, no significant difference of BMD at both sites, calcium intake and exercise was found in the early postmenopausal groups of these two areas. CONCLUSIONS: There were significant differences in BMD and lifestyle factors between late postmenopausal women in Bangkok and other provinces. Environmental factors especially calcium intake, exercise and sunlight exposure, may influence BMD in late postmenopausal Thai women.

Physical activity and risk factors for hip fractures in Thai men.

To test the hypothesis that hip fracture is associated with physical activity in Thai elderly men, a case-control study was conducted in Bangkok, Thailand. A total of 187 men aged 51 years over, resident in Bangkok, admitted consecutively with a radiologically confirmed first hip fracture were studied. 177 age-matched community controls were randomly recruited from the same neighborhood of the cases. Physical activity was independently associated with reduced risk of hip fracture after controlling for confounding factors. Very active and active past physical activity markedly reduced risk of hip fracture in comparison to subjects with inactive past physical activity. Recent active physical activity was also protective against hip fracture. This prompts a need to identify strategy to promote physical activity among the elderly and at an early age.

Bone mineral density and histology in distal renal tubular acidosis.

BACKGROUND: Chronic metabolic acidosis in distal renal tubular acidosis (RTA) has been implicated in the pathogenesis of enhanced bone resorption and osteopenia, resulting in a loss of bone mineral content. However, histomorphometric and bone densitometric studies of patients who suffered from long-standing distal RTA have rarely been done. METHODS: A cross-sectional study to determine the alterations of bone mineral density (BMD) and histology was done in 14 nonazotemic RTA patients (11 females and 3 males) who had never received alkaline therapy before enrolling into this study. The mean age was 32.7 +/- 11.9 years. BMD measurements and transiliac bone biopsy were done in all patients. Blood chemistries, intact parathyroid hormone level, and a 24-hour urine collection for the determination of urinary calcium, phosphate, sodium, and potassium were obtained from the RTA patients at the time of bone biopsy. Data from 28 age-, sex-, and body mass index-matched, normal controls who were residents in the same area were also obtained. RESULTS: Urinary excretion of calcium was 2.05 +/- 1.59 mmol/day. No patient had hypercalciuria. The serum intact parathyroid hormone level was 15.92 +/- 8.48 pg/mL. RTA patients had lower BMD in most areas when compared with normal controls. There were two patients who suffered from a pathologic fracture at the femur. Bone histomorphometry from RTA patients shows a significantly decreased bone formation rate (0.02 +/- 0.02 vs. 0.07 +/- 0.045 microm(3)/microm(2)/day, P < 0.05), not significantly decreased osteoblastic surface (0.78 +/- 1.03% vs. 2.6 +/- 1.1%) and osteoclastic surface (0.05 +/- 0.03 vs. 0.13 +/- 0.23%), but significantly increased osteoid surface (31.47 +/- 24.52 vs. 5.79 +/- 4.39%, P < 0.05) and osteoid volume (2.95 +/- 3.09 vs. 0.92 +/- 1.05%, P < 0.05) when compared with those of normal controls. There was no difference in osteoid thickness (10.65 +/- 6.10 vs. 8.69 +/- 2.14 microm). Only one distal RTA patient who had a marked increase in osteoid thickness justified the diagnosis of osteomalacia. CONCLUSIONS: This study demonstrates that low bone mass is common in distal RTA patients. Chronic metabolic acidosis results in suppression of bone formation and resorption, which in turn may contribute to the development of low bone mass in distal RTA patients. Although minor elevations in osteoid surface and osteoid volume are found among distal RTA patients, overt osteomalacia is not the predominant bone lesion.

Association of a G2014A transition in exon 8 of the estrogen receptor-alpha gene with postmenopausal osteoporosis.

We report the association of a newly identified synonymous G2014A single nucleotide polymorphism (SNP) which does not alter the amino acid sequence in exon 8 of the estrogen receptor-alpha (ERalpha) gene with osteoporosis in Thai postmenopausal women. Subjects consisted of 228 postmenopausal women aged more than 55 years divided into two groups--with vertebral or femoral osteoporosis (n = 106) or without osteoporosis (n = 122)--according to bone mineral density (BMD) criteria. The exon 8 G2014A SNP, which is 6 nucleotides upstream from the end of the stop codon, was identified by PCR-RFLP. Data are expressed as the mean and 95% CI. The allele frequency of the G2014A polymorphism was 26.4% in osteoporotic subjects and was significantly higher than that in non-osteoporotic women (15.2%) (p<0.05). By stepwise logistic regression analysis, it was found that the G2014A polymorphism was related to the presence of osteoporosis (odds ratio 2.7 per A allele, 95% CI 1.49-4.76) independently of body weight (odds ratio 0.93 per kg, 95% CI 0.89-0.96) and years since menopause (odds ratio 1.12 per year, 95% CI 1.08-1.19). In a multiple linear regression model, L2-L4 BMD of osteoporotic subjects was associated with body weight (p<0.05), endogenous estradiol levels (p<0.05) and the G2014A genotype (p<0.001), while it was related only to body weight (p<0.05) and estradiol levels in non-osteoporotic women (p<0.05). We conclude that a G2014A SNP in exon 8 of ERalpha is associated with the presence and severity of postmenopausal osteoporosis. Linkage disequilibrium between this polymorphism and the 3'-untranslated region of the ERalpha gene which may participate in the regulation of ERalpha gene expression remains to be determined.

Association of a T262C transition in exon 1 of estrogen-receptor-alpha gene with skeletal responsiveness to estrogen in post-menopausal women.

Polymorphic genetic markers of estrogen-receptor-alpha (ERalpha) gene studied so far in osteoporosis reside in non-coding region with uncertain functional significance. The purpose of the present study was to search for nucleotides changes in the exon 1 and 5' regulatory region of ERalpha gene, to study the nature of their linkages to the previously reported Pvull polymorphism in intron 1 and their functional significance in postmenopausal osteoporosis. Direct sequencing of exon 1 and promotor region of ERalpha gene revealed a synonymous nucleotide substitution from T to C at position 262, 29 nucleotides downstream from the putative start codon. No nucleotide change was found in the promotor region. Linkage disequilibrium between the T262C polymorphism and the Pvull polymorphism in intron 1 of ERalpha gene was demonstrated in 129 post-menopausal women (p<0.001). After treating 96 post-menopausal with 0.3 mg or 0.625 mg conjugated equine estrogen (CEE) for 2 yr, vertebral bone mineral density (BMD) increased regardless of the T262C genotype. However, with regard to femoral neck BMD, only those subjects that were homozygous for the T262C polymorphism had an increase in femoral BMD (+5.9+/-1.4%, mean+/-SE; p<0.0001). Using analysis of covariance to assess the effects of the T262C polymorphism, the intronic Pvull polymorphism, doses of CEE and the corresponding baseline BMD on the changes in vertebral or femoral BMD after treatments, it was found that the change in vertebral BMD was related only to the baseline BMD (p<0.05). The change in femoral BMD was independently related to the T262C polymorphism (p<0.01) and the baseline femoral BMD (p<0.01). No effect of the Pvull polymorphism or the doses of CEE on femoral BMD was demonstrated. We concluded that the previously described intronic Pvull polymorphism of ERalpha gene is in linkage disequilibrium with a T262C polymorphism in exon 1. This T262C polymorphism appears to be more directly related to the skeletal response after long-term treatment with estrogen.

Oestrogen-receptor-alpha gene polymorphism affects response in bone mineral density to oestrogen in post-menopausal women.

OBJECTIVE: An oestrogen-receptor-alpha (ERalpha) gene polymorphism has been variably reported to be related to bone mass. To investigate whether this ERalpha gene polymorphism is associated with a functional difference, we assessed the response in bone mineral density (BMD) to oestrogen therapy in post-menopausal women in relation to ERalpha gene polymorphism. PATIENTS AND MEASUREMENTS: Subjects consisted of 124 Thai post-menopausal women. Sixty-three of the women were less than 6 years post-menopausal and 61 were more than 10 years post-menopausal with vertebral or femoral osteoporosis as defined by BMD T-score less than - 2.5. Subjects were randomly allocated to receive 0.3 mg (n = 67) or 0.625 mg (n = 57) of conjugated equine oestrogen (CEE). All subjects also took 5 mg medroxyprogesterone acetate. Vertebral and femoral neck BMD were measured at baseline and 1 year after treatment. Data were expressed as mean +/- SEM. Capital P represents the absence of the restriction site while lower-case p indicates the presence of the restriction site. RESULTS: For subjects on 0.625 mg CEE, BMD at L2-4 increased significantly after 1 year in those with pp (n = 20) Pp (n = 29) and PP genotypes (n = 8) (P < 0.001). However, in subjects on 0.3 mg CEE, BMD at L2-4 increased significantly after 1 year in subjects with Pp (n = 34, + 7.6 +/- 1.5%, P < 0.001) and PP genotypes (n = 13, + 6. 9 +/- 1.0%, P < 0.001), but not in those with pp genotype (n = 20, + 2.3 +/- 2.1%, P = NS). After adjusting for age and years since menopause, the change in vertebral BMD was still lower in those without the P allele compared to those with the P allele (P < 0.05). Femoral BMD did not significantly change regardless of dose of CEE and genotype. CONCLUSIONS: We conclude that ERalpha gene polymorphism affects skeletal response to oestrogen in post-menopausal women. The effect of ERalpha gene polymorphism appears to be site-specific and does not relate to biochemical markers of bone turnover. Determination of ERalpha genotype may help identify post-menopausal women who will have more skeletal benefit from oestrogen therapy.

Bone mineral density in primary and secondary amenorrhea.

UNLABELLED: Amenorrhea in young women is one of the best clinical indicators for estrogen deficiency, except in the presence of gynecological structural pathology. This study aimed at investigating bone mineral density (BMD) in patients with primary and secondary amenorrhea. Thirty-six patients were enrolled in the study, seven with primary amenorrhea (mean age 24.3 +/- 4.5 yrs.) and twenty-nine with secondary amenorrhea (mean age 31.1 +/- 6.9 yrs.). Eighteen regularly menstruating women (mean age 31.8 +/- 3.7 yrs.) served as controls. BMD was measured at lumbar spine, femoral neck, Ward's triangle and trochanter. RESULTS: BMD was significantly decreased in both primary and secondary hypoestrogen amenorrheic patients. Primary amenorrheic patients were more severely affected with a BMD mean Z score below 80 per cent (osteopenia) at all sites measured. The age of primary amenorrheic women also strongly correlated with degree of demineralization. This should emphasize the importance of early diagnosis and treatment of young amenorrheic patients.

A T2S cross-reactive material (compound W) in hyperthyroid patients with trophoblastic disease.

In a previous study we observed increased serum levels of a 3,3'-diiodothyronine sulfate (T2S)-like material (compound W) in women who received human chorionic gonadotropin (hCG) treatment. In the present study we assessed serum compound W values in 113 women (total serum sample: 190) with trophoblastic disease, in 7 normal nonpregnant women during the menstrual cycle and 7 women given hCG treatment in the course of in vitro fertilization. Fifty-three women with trophoblastic disease had serum free thyroxine (FT4) concentrations greater than 3.0 ng/dL with suppressed serum thyrotropin (TSH) levels; 61 had FT4 values less than 3.0 ng/dL with a mean TSH of 0.83 mU/L. Mean (+/- SE) compound W concentrations in the high FT4 group were significantly higher than in the low FT4 group (76 +/- 8.1 vs. 21 +/- 1.7 ng T2S equivalent, p < 0.001) There were significant correlations between serum hCG and compound W concentrations (r = 0.472, p < 0.001), serum FT4 and hCG (r = 0.503, p < 0.0001) and serum FT4 and compound W (r = 0.585, p < 0.0001). In nonpregnant women serum compound W levels increased from 7.5 +/- 8 ng/dL at the end of the menstrual period to 15 +/- 1.7 ng/dL 21 days after the last menstrual period. Finally, a single dose of hCG (10,000 USP units, intramuscularly) increased mean (+/- SE) serum compound W levels from 12.8 +/- 2.3 to 64 +/- 9.7 ng/dL and 54 +/- 12 ng/dL at 9 and 16 days, respectively. These results indicate that hCG and perhaps luteinizing hormone (LH) increase serum compound W concentrations in women. The mechanism and significance presently are unclear.

Five-year thyrotropin screening for congenital hypothyroidism in Ramathibodi Hospital.

OBJECTIVES: To detect newborns with congenital hypothyroidism (CH) and to treat the affected infants as early as possible. STUDY DESIGN: Cord blood thyrotropin (TSH) screening for CH in Ramathibodi Hospital began in 1993. From October 1993 to December 1998, 35,390 neonates were screened. The infants with elevated TSH level of greater than 30 mU/L were recalled for verification of CH. Confirmation tests included total thyroxine, free thyroxine and TSH level. Thyroid scan and uptake were performed in some affected infants. RESULTS: Twelve infants with CH were detected resulting in an incidence of one in 2,949 live-births. All affected infants were asymptomatic at birth. Of 12 infants with CH, one premature neonate had a delayed TSH elevation and was diagnosed as having primary hypothyroidism at 2 months of age. The recall rate for validation of CH based on a cut-off value at serum TSH level of greater than 30 mU/L is 1.1 per cent. If the cut-off value of serum TSH level was raised to greater than 40 mU/L, the recall rate would decrease to 0.43 per cent. None of the affected infants had cord blood TSH level of less than 50 mU/L except one premature patient. Therefore, beginning in January 1997, the cut-off value of TSH was raised to 40 mU/L or greater. Pitfalls in this program include incomplete blood-specimen collection and incomplete follow-up. To strengthen the program, improvements were made in the follow-up system from 1996 onward. Therefore, the coverage for blood-specimen collection progressively increased from 84 per cent in 1994 to 96 per cent in 1998. Simultaneously, the patients' return after recalls also increased from 38 per cent to 100 per cent. CONCLUSIONS: The incidence of CH in Ramathibodi Hospital is approximately 1:3,000 live-births. The optimal cord blood TSH level for recall is 40 mU/L or greater. The intensification of follow-up strategy resulted in better response to recall and earlier treatment in the affected infants.

Estrogen receptor gene polymorphism is associated with bone mineral density in premenopausal women but not in postmenopausal women.

In the present study, we examined the genotypes distribution of Pvu II estrogen receptor (ER) gene polymorphism and its association to bone mass in Thai females. Subjects consisted of 134 Thai females 54 of whom were premenopausal and 80 were postmenopausal. Pvu II ER gene polymorphism was determined by PCR-RFLP. Capital P represents the absence of the restriction site while small p indicates the presence of the restriction site. Forty nine (36.6%) of the subjects had pp genotype, while 59 (44.0%) had Pp genotype and 26 (19.4%) had PP genotype. There was no significant difference in age, body weight, height and calcium intake in premenopausal women with different genotypes. The results including years since menopause were similar in postmenopausal women. When including ER gene genotypes, age, body weight, height and dietary calcium intake in a stepwise multiple regression model, it was found that besides body weight ER gene polymorphism was associated with bone mineral density (BMD) at AP spine (p < 0.05), lateral spine (p < 0.05) femoral neck (p < 0.05) and femoral trochanter (p < 0.05) with the pp genotype having the least BMD. ER gene polymorphism was the only factor associated with BMD at Ward's triangle, (p < 0.05) while only body weight was associated with BMD at distal and mid radius. There was no difference in serum intact osteocalcin (OC) concentrations among subjects with different genotypes. ER gene polymorphism was not related to BMD in postmenopausal women at any skeletal site. Similarly, serum intact OC levels were not different among postmenopausal women with different genotypes. We concluded that Pvu II estrogen receptor gene polymorphism is associated with bone mineral density in premenopausal women but not in postmenopausal women. Estrogen receptor gene polymorphism may have a modulatory role in calcium and bone metabolism during adolescence and young adulthood.

Serum oestradiol and oestrogen-receptor gene polymorphism are associated with bone mineral density independently of serum testosterone in normal males.

OBJECTIVES: The physiological effects of oestrogens on bone in men were largely unanticipated until recently, when oestrogen deficiency in males with aromatase deficiency and oestrogen resistance was found to cause osteoporosis and delayed fusion of epiphyses despite sufficient serum testosterone. This raises the possibility that in normal men oestrogens rather than androgens are of physiological importance in bone maturation. In the present study, we examined the association of serum oestradiol (E2) compared to that of free testosterone (FT) with bone mineral density (BMD) in normal men. The effect of oestrogen receptor (ER) gene polymorphism on BMD in men was also addressed. SUBJECTS: Eighty-one Thai men aged 20-79 years. All were healthy and did not take medication which may affect calcium and bone metabolism. BMD was assessed by DEXA. Dietary calcium was assessed by a 3-day dietary record. Serum E2 and FT concentrations were measured by radioimmunoassay. Polymorphism at intron 1 of the alpha isoform of ER gene was determined by PCR-RFLP. Small p represents the presence of the restriction site while capital P indicates the absence of the restriction site. RESULTS: Serum FT decreased with increasing age (r = -0.58, P < 0.0001) while E2 did not. However, there was a positive association between E2 and FT (r = 0.28, P < 0.05). Serum FT was related to BMD at femoral neck (r = 0.26, P < 0.05) and Ward's triangle (r = 0.30, P < 0.01) while E2 was related to BMD at anteroposterior (AP) lumbar spine (r = 0.29, P < 0.05), femoral neck (r = 0.23, P < 0.05) and femoral trochanter (r = 0.27, P < 0.05). Besides FT and E2, age, body weight, fat mass and fat-free mass were also correlated to BMD at various skeletal sites. Using stepwise multiple linear regression to control for the confounding effects among these factors, fat-free mass was found to be strongly associated with BMD at most skeletal sites. Serum E2 was related to BMD independently of other factors including FT at AP lumbar spine (r = 0.22, P < 0.05), femoral neck (r = 0.26, P < 0.01), femoral trochanter (r = 0.22, P < 0.05) and Ward's triangle (r = 0.26, P < 0.01) while serum FT was not associated with BMD at any site after controlling for E2 and other related factors. Concerning ER alpha gene polymorphism, 27 (33.3%) of the subjects had pp genotype, while 42 (51.9%) and 12 (14.8%) Pp and PP genotypes, respectively. After controlling for age, body weight, fat mass, fat-free mass, calcium intake, FT and E2, the presence of P allele was associated with higher BMD at AP L2-L4 (P < 0.05). CONCLUSIONS: Serum oestradiol is more related to bone mass than free testosterone in normal men. Oestrogen-receptor gene polymorphism is also associated with bone mass in men independently of oestradiol levels. Serum oestradiol together with oestrogen-receptor genotype may partly determine bone mass in males.

The effects of oestrogen exposure on bone mass in male to female transsexuals.

OBJECTIVE: The importance of oestrogen on bone mineral density (BMD) in males was suggested by reports of patients with oestrogen resistance and aromatase deficiency who demonstrated osteoporosis and epiphyseal plate maturation defect despite high testosterone levels. In the present study, we examined the effects of oestrogen exposure on BMD in transsexual men. DESIGN: Cross-sectional study of BMD in male to female transsexuals. PATIENTS: Subjects consisted of two groups of transsexual male dancers aged 16-34 years who did not receive transsexual operations (n = 28). Group 1 (n = 11) and group 2 (n = 17) had used oestrogen for 2 years or less and more than 2 years, respectively. Twenty-four healthy adult males served as controls. RESULTS: Signs of feminization were presented in both group 1 and group 2, with Tanner's stage II-III breast development. BMD at various sites were correlated only to body weight and not to smoking or milk consumption. After controlling for body weight, it was found that group 2 had significantly higher BMD at L2-4 than controls (1.22 +/- 0.03 vs. 1.14 +/- 0.03 g/cm2, P < 0.05) and group 1 (1.22 +/- 0.03 vs. 1.08 +/- 0.04 g/cm2, P < 0.05). BMD at femoral neck was also higher in group 2 compared to controls (1.10 +/- 0.03 vs. 1.01 +/- 0.03 g/cm2, P < 0.05) and group 1 (1.10 +/- 0.03 vs. 0.95 +/- 0.04 g/cm2, P < 0.05). Group 1 subjects had lower BMD compared to controls at femoral trochanter (0.70 +/- 0.04 vs. 0.83 +/- 0.03 g/cm2, P < 0.05) and total femur (0.96 +/- 0.05 vs. 1.07 +/- 0.03 g/cm2, P < 0.05). CONCLUSIONS: Long-term oestrogen exposure transsexual men result in an increase in bone mineral density despite signs of feminization. This suggests that oestrogen has positive effects on bone density in males. The finding of the trend towards reduced bone density in group 1 remains unexplained.

Preevacuation serum CA 125 in complete hydatidiform mole.

Studies on CA 125 in hydatidiform mole are limited. The objective of this study was to measure the preevacuation serum CA 125 level in patients with complete hydatidiform mole and to determine whether it could predict the later development of persistent trophoblastic disease. Preevacuation serum CA 125 levels were immunoradiometrically measured in 69 patients with histologically confirmed complete hydatidiform mole. The mean (range) serum CA 125 level was 63.7 (10.5-404.7) U/ml. Using 35 U/ml as the cutoff point, the elevated CA 125 levels were observed in 53.6% (37/69) of the patients. The mean serum CA 125 level of patients who later developed persistent trophoblastic disease was not significantly higher than that of those who had benign course (78.9 vs 52.6 U/ml, P > 0.05). In conclusion, the preevacuation serum CA 125 level was elevated in about half of patients with complete hydatidiform mole and it could not be used to predict the subsequent development of persistent trophoblastic disease.

Evaluation of five commercial kits for free thyroxine estimation by comparison with equilibrium dialysis method in nonthyroidal illness.

Five commercial kits for estimating FT4 in serum of 59 euthyroid control and 38 patients with severe NTI were studied: one non analog method (Gammacoat two step RIA, Clinical Assay) and four different analog methods (Amerlex-M RIA, Amersham; Enzymun test competitive enzyme immunoassay, Boehringer Mannheim; Amerlite chemiluminescence immunoassay, Kodak Clinical Diagnostics; Berilux chemiluminescence immunoassay, Behring) compared with equilibrium dialysis (Eq) method. Serum FT4 estimates in NTI patients measured by all commercial kits in this study yielded results comparable with those by equilibrium dialysis. The proportions of serum FT4 values concordant with FT4 (Eq) in each kit were 76.3, 76.3, 76.3, 68.4 and 78.9 per cent respectively. The percentage of NTI patients who had serum FT4 values lower than the reference levels of the methods used were 21.2, 26.3, 7.9, 15.8, 18.4 and 18.4 per cent respectively. No patient with low serum FT4 (Eq) level had subnormal or high serum TSH value. However, 4 out of 6 patients with high serum FT4 (Eq) values had depressed serum TSH values. All of them also had elevation of serum FT4 estimates measured by all kits. Serum FT4 estimates measured by all methods correlated well with FT4 (Eq) levels within the NTI group.

Serum testosterone and its relation to bone mineral density and body composition in normal males.

OBJECTIVE: Bone mineral density (BMD) declines with age in both men and women, predisposing the elderly to osteoporosis and fractures. Although there are extensive data about post-menopausal osteoporosis, there is relatively little information concerning the decrease in BMD with age in normal men, particularly the contribution of declining gonadal function with age to BMD. In the present study, we investigated the effect of age on the pituitary-gonadal axis in normal males and its relation to BMD and body composition. SUBJECTS: Ninety healthy Thai males in the Bangkok Metropolitan area without a history of smoking or significant alcohol consumption were studied. MEASUREMENTS: Serum testosterone (T), free testosterone (FT), LH and FSH were measured by radioimmunoassay in fasting blood samples obtained in the morning between 0600 and 1000 h. BMD at anteroposterior L2-L4, lateral L2-L4, femoral neck, femoral trochanter and Ward's triangle were determined by dual-energy X-ray absorptiometry. RESULTS: There were significant declines with age in BMD at lateral L2-L4 (r = -0.37, P < 0.001), femoral neck (r = -0.49, P < 0.0001), Ward's triangle (r = -0.54, P < 0.0001) but not at anteroposterior L2-L4 or femoral trochanter. Serum FT (r = -0.56, P < 0.0001) but not T (r = -0.19, P = 0.07) decreased with age. Serum LH (r = 0.27, P < 0.001) and FSH (r = 0.4, P < 0.0001) increased with age suggesting a defect in gonadal androgen synthesis or possibly a secretion of bioinactive LH. Serum FT concentrations were significantly correlated to lateral L2-L4 (r = 0.27, P < 0.05), femoral neck (r = 0.48, P < 0.0001) and Ward's triangle (r = 0.50, P < 0.0001) BMD. After controlling for age, declining FT with age was still associated with a decrease in BMD in femoral neck (P < 0.05) and Ward's triangle (P < 0.05) but not in lateral L2-L4. The proportion of body fat increased with age (r = 0.3, P < 0.01). Decreased serum T, but not FT, was associated with increased body fat after age was taken into account (P < 0.0001). CONCLUSIONS: There is a decline in serum free testosterone together with increases in LH and FSH with age in healthy males. The decrease in serum free testosterone is partially associated with the age-related decline in bone mineral density added to the effect of age at the femoral neck and Ward's triangle. Testosterone but not free testosterone is associated with age-related increase in body fat.

Virilization associated with choriocarcinoma.

We demonstrate here two autopsy-proven cases of virilization associated with choriocarcinoma. The first patient was a 27-year-old woman with a 7-year history of metastatic choriocarcinoma who presented with left hemiparesis and virilization. Serum testosterone concentration was 18 nmol/l, free testosterone 471 pmol/l, dehydroepiandrosterone sulphate (DHEA-sulphate) 1.7 mumol/l, sex hormone binding globulin 12.3 nmol/l, estradiol (E2) 1,843 pmol/l, and hCG 1.6 x 10(5) IU/l. The second patient was a 21-year-old virilized woman with metastatic choriocarcinoma who presented in semicomatose state. Limited endocrine investigation revealed serum testosterone 27 nmol/l and hCG 2.7 x 10(5) IU/l. Both patients died despite chemotherapy and radiation therapy. Autopsy findings revealed choriocarcinoma with brain and multiple organ metastasis in both. Pathology of the ovaries of both patients demonstrated hyperplasia of luteinized theca cells and lutein cells. We postulate that an association of virilization and choriocarcinoma resulted from long-standing stimulation of ovary by hCG causing theca cell hyperplasia with subsequent hypertestosteronemia and virilization.