Omar M. Hauache

Vitamin D endocrine system and the genetic susceptibility to diabetes, obesity and vascular disease. A review of evidence

The Vitamin D endocrine system regulates multiple aspects of calcium metabolism and cellular differentiation and replication in the immune system, endocrine pancreas, liver, skeletal muscles and adipocytes. It plays an important role in glucose homeostasis, notably, in the mechanism of insulin release. Actions of vitamin D are mediated by the binding of 1, 25-(OH)2D3 to a specific cytosolic/nuclear vitamin D receptor (VDR), a member of the steroid/thyroid hormone receptor superfamily. Several frequent polymorphisms are found in the VDR gene and were reported to be associated with a variety of physiological and pathological phenotypes in many populations. In this paper, we will review the evidences suggesting associations of allelic variations in the VDR gene and phenotypes related to body weight, glucose homeostasis, diabetes and its vascular complications.

Screening for macroprolactinaemia and pituitary imaging studies

objective Hyperprolactinaemia is caused by high levels of monomeric, dimeric or macro forms of prolactin in circulation, the monomeric form being predominant in patients with prolactinomas. Macroprolactinaemia, however, is common and is associated with asymptomatic cases. In this study, we reviewed our records regarding clinical and imaging investigations in patients who were found to have hyperprolactinaemia predominantly due to the presence of macroprolactin and compared them with the findings observed in patients whose prolactin molecular size consisted predominantly of the monomeric form.

Extracellular calcium-sensing receptor: structural and functional features and association with diseases

The recently cloned extracellular calcium-sensing receptor (CaR) is a G protein-coupled receptor that plays an essential role in the regulation of extracellular calcium homeostasis. This receptor is expressed in all tissues related to this control (parathyroid glands, thyroid C-cells, kidneys, intestine and bones) and also in tissues with apparently no role in the maintenance of extracellular calcium levels, such as brain, skin and pancreas. The CaR amino acid sequence is compatible with three major domains: a long and hydrophilic aminoterminal extracellular domain, where most of the activating and inactivating mutations described to date are located and where the dimerization process occurs, and the agonist-binding site is located, a hydrophobic transmembrane domain involved in the signal transduction mechanism from the extracellular domain to its respective G protein, and a carboxyterminal intracellular tail, with a well-established role for cell surface CaR expression and for signal transduction. CaR cloning was immediately followed by the association of genetic human diseases with inactivating and activating CaR mutations: familial hypocalciuric hypercalcemia and neonatal severe hyperparathyroidism are caused by CaR-inactivating mutations, whereas autosomal dominant hypoparathyroidism is secondary to CaR-activating mutations. Finally, we will comment on the development of drugs that modulate CaR function by either activating (calcimimetic drugs) or antagonizing it (calcilytic drugs), and on their potential therapeutic implications, such as medical control of specific cases of primary and uremic hyperparathyroidism with calcimimetic drugs and a potential treatment for osteoporosis with a calcilytic drug.

Vitamin D Receptor Gene Polymorphism: Correlation with Bone Mineral Density in a Brazilian Population with Insulin-Dependent Diabetes Mellitus

Abstract: Patients with insulin-dependent diabetes mellitus (IDDM) are at higher risk of developing osteoporosis. Among the genetic factors related to the development of osteoporosis, a possible association between vitamin D receptor (VDR) gene polymorphism and bone mineral density (BMD) has been described in some populations. We characterized the VDR gene polymorphism in a healthy adult Brazilian population and in a group of patients with IDDM and correlated these findings with densitometric values in both groups. The Brazilian population is characterized by an important racial heterogeneity and therefore is considered an ethnically heterogeneous population. We recruited 94 healthy adult Brazilian volunteers (63 women and 31 men), mean (+ SD) age 32.4 + 6.5 years (range 18–49 years), and 78 patients with IDDM (33 women and 45 men) diagnosed before 18 years of age, mean (+ SD) age 23.3 + 5.5 years (range 18–39 years). VDR genotype was assessed by polymerase chain reaction amplification followed by BsmI digestion on DNA isolated from peripheral blood leukocytes. Statistical analysis included Bonferroni t-test to compare densitometric values within different genotypes in both groups and multiple regression analysis of bone density adjusted for potential confounding factors. The IDDM group had a lower BMD compared with the control group. The VDR genotype distribution in the control group was 43 Bb (45.7%), 39 bb (41.5%) and 12 BB (12.8%). This distribution did not differ from that observed in the IDDM group: 39 Bb (50%), 26 bb (33.3%) and 13 BB (16.7%). In the IDDM group, patients with the Bb genotype had a higher body weight when compared with the BB genotype (p= 0.02). However, when diabetic patients were controlled for age, sex and body mass index, BB genotype was associated with a lower mean BMD at lumbar spine and femoral neck than in Bb and bb patients. BB patients had a shorter duration of IDDM than bb and Bb patients. These findings suggest a small influence of VDR gene polymorphism on BMD of a racially heterogeneous population with IDDM.

Body fat and cholecalciferol supplementation in elderly homebound individuals

Vitamin D deficiency, observed mainly in the geriatric population, is responsible for loss of bone mass and increased risk of bone fractures. Currently, recommended doses of cholecalciferol are advised, but since there are few studies evaluating the factors that influence the serum levels of 25-hydroxyvitamin D (25(OH)D) following supplementation, we analyzed the relationship between the increase in serum 25(OH)D after supplementation and body fat. We studied a group of 42 homebound elderly subjects over 65 years old (31 women) in order to assess whether there is a need for adjustment of the doses of cholecalciferol administered to this group according to their adipose mass. Baseline measurements of 25(OH)D, intact parathyroid hormone and bone remodeling markers (osteocalcin and carboxy-terminal fraction of type 1 collagen) were performed. Percent body fat was measured by dual-energy X-ray absorptiometry. The patients were divided into three groups according to their percent body fat index and were treated with cholecalciferol, 7,000 IU a week, for 12 weeks. The increases in serum levels of 25(OH)D were similar for all groups, averaging 7.46 ng/mL (P < 0.05). It is noteworthy that this increase only shifted these patients from the insufficiency category to hypovitaminosis. Peak levels of 25(OH)D were attained after only 6 weeks of treatment. This study demonstrated that adipose tissue mass does not influence the elevation of 25(OH)D levels following vitamin D supplementation, suggesting that there is no need to adjust vitamin D dose according to body fat in elderly homebound individuals.

Distribution of HOMA-IR in Brazilian subjects with different body mass indexes.

N THE CLINICAL SETTING, assessment of insulin resistance (IR) can be made without any laboratory test, but by evaluating data such as weight or BMI, among others. However, in the last few years laboratories have been asked to provide direct or indirect measures of IR. Homeostatic model assessment (HOMA-IR) is a simple and inexpensive technique to evaluate IR from basal (fasting) glucose and insulin or C-peptide concentrations, validated against a variety of physiological methods (1-4). To validate the potential use of HOMA-IR values in clinical practice, it is important to establish its distribution in a sample of normoglycemic individuals from a population with the same genetic background (2). In this regard, we aim to describe the distribution of HOMA-IR values in a large sample of Brazilian population. We analyzed retrospectively fasting insulin and glucose levels in 1,898 Brazilian adults (25% males, age range 18-90 years). All had fasting glucose 30 (n= 391) (table 1). A recent publication in a group of 312 non-obese, normoglicemic Brazilian subjects found a mean HOMA-IR of 1.7, and a threshold value for insulin resistance of 2.71 (3). The use of HOMA ‐IR to quantify insulin sensitivity can be helpful in clinical practice for comparisons of insulin sensitivity, gathering of longitudinal data in subjects at risk for development of abnormal glucose tolerance. Also, HOMA ‐IR can be used to assess longitudinal changes of IR in patients with diabetes in order to examine the natural history of the disease and to assess the effects of treatment, as demonstrated for example in the U.K. Prospective Diabetes Study (UKPDS) (5). Our results can be useful as a reference for further Brazilian studies using HOMA-IR values as a measure of IR in this particular population.

A novel mutation in the LRP5 gene is associated with osteoporosis-pseudoglioma syndrome

Dear Editor, Osteoporosis-pseudoglioma syndrome (OPPG) is an autosomal recessive disorder characterized by low bone mass and ocular globe alterations, with frequent fractures as a consequence of high skeletal fragility and severe visual deficiency [1]. Recently, Gong et al. identified mutations in the low-density lipoprotein receptor-related protein 5 gene (LRP5) that cause OPPG [2]. The LRP5 gene is located on the long arm of chromosome 11 at 11q13.4, has 23 exons, and encodes a transmembrane protein with 1,615 amino acids. This protein is a member of the low-density lipoprotein receptor (LDLR) family, the members of which have very well conserved characteristics. We had the opportunity to follow-up two brothers with clinical characteristics of OPPG, and we decided to investigate the LRP5 gene to identify any mutation that could explain the clinical features of low bone mass, fractures, and blindness. The older brother came to the hospital when he was 12 years old, (53 kg; 157 cm). At that time, lumbar bone mineral density (BMD, Hologic 4,500 A) was 0.512 g/cm (Z score −3.99). He was born blind, had a normal development until the age of 7 years, when he had lumbar pain due to the fracture of a vertebral body. At the age of 11, pain in the left leg led to the diagnosis of a tibial fracture. One month later, the same occurred in the right tibia and 2 months after that, a fracture in the right femur was detected, and he was referred to our hospital. The younger brother was 4 years old at that time and was born blind. At 2.5 years, he had a fracture of the right femur after a small fall. He was eutrophic (13 kg and 97 cm), with a lumbar BMD of 0.277 g/cm2 (Z score −5.4). Both of them had normal biochemical and hormonal parameters. Their parents were consanguineous, had normal BMD and no history of fractures or blindness, and there were no similar cases in the family. The 23 exons were analyzed and we identified an LPR5 missense mutation in the third codon of exon 8, in which an asparagine is replaced by isoleucine (N531I) in two siblings with OPPG and their consanguineous parents. The patients were found to be homozygous for this mutation, while their parents are heterozygous. This amino acid is located at the second propeller domain (YWTD repeat) in the extracellular domain of the receptor. Fifty normal participants underwent analysis of this exon, and the mutation was not present in any of them, but a polymorphism C→T (rs545382 NCBI, F549F) was found at a frequency of 1:5. According to Sidow’s program [3] of multivariate analysis of protein polymorphism (MAPP), we observed that the N531I mutant scored higher (yielding a bad amino acid substitution) and this mutation is located in the group of deleterious variants because of its high degree of evolutionary conservation. Based on these facts, we believe that this mutation is responsible for the clinical features presented in our patients and thereby define a novel mutation that causes OPPG in humans. Osteoporos Int DOI 10.1007/s00198-007-0360-x

Serum leptin concentration during puberty in healthy nonobese adolescents

Data obtained during the past five years have indicated that there are important age- and gender-based differences in the regulation and action of leptin in humans. To study the physiological changes of leptin during puberty in both sexes, and its relationship with body composition and sexual maturation, we measured leptin concentrations in 175 healthy adolescents (80 girls, 95 boys, 10-18 years of age), representing all pubertal stages. We excluded individuals with a body mass index (BMI) below the 5th or above the 95th percentile relative to age. Serum concentrations of leptin were determined by a monoclonal antibody-based immunofluorimetric assay, developed in our laboratory. Body composition was determined by dual-energy X-ray absorptiometry. Pubertal stage was assigned by physical examination, according to Tanner criteria for breast development in females and genital development in males. Leptin concentration in girls (N = 80) presented a positive linear correlation with age (r = 0.35, P = 0.0012), BMI (r = 0.65, P < 0.0001) and %fat mass (r = 0.76, P < 0.0001). In boys (N = 95) there was a positive correlation with BMI (r = 0.49, P < 0.0001) and %fat mass (r = 0.85, P < 0.0001), but a significant negative linear correlation with Tanner stage (r = -0.45, P < 0.0001) and age (r = -0.40, P < 0.0001). The regression equation revealed that %fat mass and BMI are the best parameters to be used to estimate leptin levels in both sexes. Thus, the normal reference ranges for circulating leptin during adolescence should be constructed according to BMI or %fat mass to assure a correct evaluation.

Changes in clinical and laboratory findings at the time of diagnosis of primary hyperparathyroidism in a University Hospital in São Paulo from 1985 to 2002

In contrast to most developed countries, most patients with primary hyperparathyroidism in Brazil are still symptomatic at diagnosis. However, we have been observing a change in this pattern, especially in the last few years. We evaluated 104 patients, 77 females and 27 males aged 11-79 years (mean: 54.4 years), diagnosed between 1985 and 2002 at a University Hospital. Diagnosis was made on the basis of clinical findings and of high total and/or ionized calcium levels, high or inappropriate levels of intact parathyroid hormone and of surgical findings in 80 patients. Patients were divided into three groups, i.e., patients diagnosed from 1985 to 1989, patients diagnosed from 1990 to 1994, and patients diagnosed from 1995 to 2002. The number of new cases diagnosed/year increased from 1.8/year in the first group to 6.0/year in the second group and 8.1/year in the third group. The first group comprised 9 patients (mean serum calcium +/- SD, 13.6 +/- 1.6 mg/dl), 8 of them (88.8%) defined as symptomatic. The second group comprised 30 patients (mean calcium +/- SD, 12.2 +/- 1.63 mg/dl), 22 of them defined as symptomatic (73.3%). The third group contained 65 patients (mean calcium 11.7 +/- 1.1 mg/dl), 34 of them symptomatic (52.3%). Patients from the first group tended to be younger (mean +/- SD, 43.0 +/- 15 vs 55.1 +/- 14.4 and 55.7 +/- 17.3 years, respectively) and their mean serum calcium was significantly higher (P < 0.05). All of symptomatic patients independent of group had higher serum calcium levels (12.4 +/- 1.53 mg/dl, N = 64) than asymptomatic patients (11.4 +/- 1.0 mg/dl, N = 40). Our data showed an increase in the percentage of asymptomatic patients over the years in the number of primary hyperparathyroidism cases diagnosed. This finding may be due to an increased availability of diagnostic methods and/or to an increased awareness about the disease.

Bone mineral density in young women of the city of São Paulo, Brazil: correlation with both collagen type I alpha 1 gene polymorphism and clinical aspects.

Osteoporosis is a multifactorial disease with great impact on morbidity and mortality mainly in postmenopausal women. Although it is recognized that factors related to life-style and habits may influence bone mass formation leading to greater or lower bone mass, more than 85% of the variation in bone mineral density (BMD) is genetically determined. The collagen type I alpha 1 (COLIA1) gene is a possible risk factor for osteoporosis. We studied a population of 220 young women from the city of São Paulo, Brazil, with respect to BMD and its correlation with both COLIA1 genotype and clinical aspects. The distribution of COLIA1 genotype SS, Ss and ss in the population studied was 73.6, 24.1 and 2.3%, respectively. No association between these genotypes and femoral or lumbar spine BMD was detected. There was a positive association between lumbar spine BMD and weight (P<0.0001), height (P<0.0156), and body mass index (BMI) (P<0.0156), and a negative association with age at menarche (P<0.0026). There was also a positive association between femoral BMD and weight (P<0.0001), height (P<0.0001), and BMI (P<0.0001), and a negative correlation with family history for osteoporosis (P<0.041). There was no association between the presence of allele s and reduced BMD. We conclude that a family history of osteoporosis and age at menarche are factors that may influence bone mass in our population.