Lilian Fukusima Hayashi

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.

Seasonal variation in the serum 25-hydroxyvitamin D levels of young and elderly active and inactive adults in Sao Paulo, Brazil: The Sao PAulo Vitamin D Evaluation Study (SPADES).

Objective: To evaluate the 25-hydroxyvitamin D [25(OH)D] concentrations in individuals in the city of São Paulo belonging to different age groups and exhibiting specific behavioral characteristics and to correlate the 25(OH)D concentration with the level of UV radiation (UVR). Patients and Methods: A total of 591 individuals were included, distributed as follows: 177 were living in institutions (NURSING, 76.2 ± 9.0 y old), 243 were part of the community elderly (COMMUNITY, 79.6 ± 5.3 y old), 99 were enrolled in a physical activity program targeting the elderly (ACTIVE, 67.6 ± 5.4 y old) and 72 were young (YOUNG, 23.9 ± 2.8 y old). Blood samples from all individuals were collected throughout the year. UVR measurements were taken by an official meteorology institution. Results: The UVR values varied throughout the year, following a sinusoidal-like pattern. Because of the Earth’s orbit, we hypothesized that there would be cyclic patterns for the 25(OH)D and UVR values that repeat every 12 mo. The general formula is represented by the equation P1+P2⋅sin(−2⋅π12⋅(t−P3)) The mean 25(OH)D concentration and the amplitude of the variation were significantly higher for the YOUNG and ACTIVE groups than for the COMMUNITY and NURSING groups. The nadir for UVR was in June, whereas the nadir for the 25(OH)D concentration was in the spring, corresponding to a delay of one season. Conclusions: There was seasonal variation in the 25(OH)D concentration for all the groups studied; however, the amplitude of the variation was higher for the groups of young and physically active people, possibly due to the higher level of sunlight exposure for these groups. The lowest 25(OH)D concentration was detected in the spring.

Increases in summer serum 25-hydroxyvitamin D (25OHD) concentrations in elderly subjects in São Paulo, Brazil vary with age, gender and ethnicity

BackgroundHypovitaminosis D is a common condition among elderly individuals in temperate-climate countries, with a clear seasonal variation on 25 hydroxyvitamin D levels, increasing after summer and decreasing after winter, but there are few data from sunny countries such as Brazil. Many factors can interfere on vitamin D cutaneous synthesis. We aimed at studying the 25OHD variations during winter and summer in an outdoor physically active elderly population living in São Paulo city, and analysed their determining factors.MethodsNinety-nine individuals (52 women and 47 men, from 55 to 83 years old) from different ethnic groups were selected from an outdoor physical activity group. Data are reported as Mean ± SD, and we used Pearson Linear Correlation, Students t-test for non-related samples, Chi-square (χ²) test and One-way ANOVA for analysis.ResultsMean 25OHD value for the whole group was 78.9 ± 30.9 nmol/L in the winter and 91.6 ± 31.7 nmol/L in the summer (p = 0.005). Mean winter serum 25OHD concentrations were not different between men and women (81.2 ± 30.1 nmol/L vs. 76.7 ± 31.8 nmol/L, respectively), and 19.2% of the individuals showed values < 50 nmol/L. In the summer, we noticed an increase only for men (107.6 ± 31.4 nmol/L) compared to women (76.7 ± 24.0 nmol/L), and 6.5% showed values < 50 nmol/L. A decrease in the mean PTH in the summer compared to the winter was noticed, with PTH levels showing a relationship with 25OHD concentrations only in the winter (r = -0.208, p = 0.041). White individuals showed an increase in mean serum 25OHD in the summer (p = 0.016) which was not noticed for other ethnic groups (Asians, native Brazilians and blacks). An increase in 25OHD values in the summer was observed in the age groups ranging from 51-60 and 61-70 years old (p < 0.05), but not in the age group from 71 years old on.Conclusions25OHD values increased during the summer in elderly residents of São Paulo, but to different extents depending on ethnicity, gender and age. This season-dependent increase was noticed only among men, white and who were in the youngest group of individuals.

Salivary cortisol as an indicator of adrenocortical function in healthy infants, using massage therapy.

CONTEXT AND OBJECTIVE The evaluation of adrenocortical function with the use of therapeutic massage has been little studied in Brazil. The purpose of this study was to evaluate the salivary cortisol levels before and after Shantala massage therapy on healthy infants. DESIGN AND SETTING Prospective case series, in a public nursery, in São Paulo. METHODS Saliva was obtained from 11 infants at the times of 8:00-9:00 a.m. and 4:00-5:00 p.m. in a nursery and 9:00-10:00 p.m. at home. They received a 15-minute therapeutic massage on two consecutive days, and saliva was collected before and after the massage. The procedure was repeated after a one-week interval. Cortisol values (intra-assay < 5%; inter-assay < 10%) at different times of the day were compared by ANOVA. RESULTS The mean cortisol values (nmol/l +/- SD) on the first day were: morning (M) = 14.1 +/- 5.7, afternoon (A) = 8.3 +/- 2.7, night (N) = 3.3 +/- 1.1; after two consecutive days of therapeutic massage: M = 22.3 +/- 13.5, A = 13.4 +/- 6.0, N = 5.8 +/- 3.5; after a one-week interval: M = 15.8 +/- 7.7, A = 14.3 +/- 7.7, N = 3.4 +/- 2.0. CONCLUSION There was a modification in the salivary cortisol values following massage, thus reflecting possible adaptation of the hypothalamic-pituitary-adrenal axis.

Psychological stress as a risk factor for postoperative keloid recurrence

OBJECTIVE To investigate psychological stress on the prognosis of the postoperative recurrence of keloids. METHODS Patients with keloids (n=25), candidates for surgical resection and postoperative radiotherapy, had their psychological stress evaluated on the day before the surgical procedure. The parameters evaluated were pain and itching (Visual Numerical Scale), quality of life (Questionnaire QualiFibro/Cirurgia Plástica-UNIFESP), perceived stress (Perceived Stress Scale), depression and anxiety (Hospital Depression and Anxiety Scale), salivary cortisol and minimum and maximum galvanic skin responses (GSR) at rest and under stress (i.e., while the questionnaires were being filled out). Patients were evaluated during the 3rd, 6th, 9th and 12th months of postoperative care. During each return visit, two experts classified the lesions as non-recurrent and recurrent. RESULTS The recurrence group presented the greatest values in GSR during a stressful situation. The chance of recurrence increased by 34% at each increase of 1000 arbitrary units in maximum GSR during stress. CONCLUSION Psychological stress influenced the recurrence of keloids.

Association between vitamin D serum levels and adolescent idiopathic scoliosis

Background Idiopathic scoliosis (IS) is a deformity of the spine that occurs in up to 4% of children during childhood and adolescence. Idiopathic scoliosis is considered multifactorial, and family history may present several individuals affected. We still cannot determine which curves will worsen and at what rate, but some factors, such as age, growth potential and skeletal maturity have been associated with a higher risk of progression. Studies have suggested that a decrease in bone mineral density may be responsible for the appearance and progression of the disease, and some have tried to link vitamin D receptor gene (VDRG) polymorphism to adolescent idiopathic scoliosis (AIS).

Association between vitamin d levels and adolescent idiopathic scoliosis

Objetivo: O objetivo deste estudo e comparar os niveis de 25-hidroxivitamina D em pacientes com EIA e com um grupo controle. Metodos: Os pacientes foram recrutados no ambulatorio de nossa instituicao durante 2013. Criancas menores de 10 ou maiores de 18 anos, portadores de disturbios ​​neurologicos ou musculares, deformidades congenitas ou sindromes geneticas foram excluidos. A dosagem de calcio, fosforo, creatinina, ureia e hormonio da paratireoide humano foi realizada para descartar disfuncoes renais ou das paratireoide. Os niveis de 25-OHD foram determinados por teste de eletroquimioluminescencia totalmente automatizado apos oito horas de jejum, durante a primavera brasileira. O nivel adequado de 25OHD foi definido como acima de 30 ng/mL. Os resultados foram comparados a um grupo de individuos saudaveis. Resultados: No grupo 1 (controle), 63,3 % apresentaram niveis anormais de vitamina D , enquanto que 91% dos pacientes portadores de EIA apresentou baixo nivel de vitamina D. A media do IMC foi de 19,6 kg/m2 para os controles e 20,3 kg/m2 no grupo 2. A analise estatistica demonstrou diferenca relevante (p < 0,0001) entre os niveis de vitamina D. A media e os valores minimos de vitamina D foram respectivamente 27 e 13,6 ng / mL para o grupo 1 e 18,8 e 3,13 ng / ml para o grupo da EIA. Conclusoes: Pacientes portadores de EIA mostraram niveis de 25OHD mais baixos do que pacientes saudaveis, com significância estatistica. O presente estudo sugere que a deficiencia da vitamina D possa estar relacionada com a etiologia da escoliose idiopatica do adolescente, corroborando dados da literatura. Mais pesquisas devem ser desenvolvidas para investigar o impacto real dos niveis sericos de vitamina D sobre a fisiopatologia da EIA.Objetivo: Comparar os níveis de 25-hidroxivitamina D em pacientes com EIA e um grupo controle. Métodos: Os pacientes foram recrutados no ambulatório de nossa instituição durante 2013. Crianças com escoliose diagnosticada antes de 10 ou após 18 anos de idade, portadoras de distúrbios neurológicos ou musculares, deformidades congênitas ou síndromes genéticas foram excluídas. Os níveis de 25-OHD foram determinados por teste de eletroquimioluminescência totalmente automatizado. O nível adequado de 25-OHD foi definido como acima de 30 ng/ml. Os resultados foram comparados a um grupo de indivíduos saudáveis. Resultados: No grupo 1 (controle), 63,3 % apresentaram níveis anormais de vitamina D, enquanto 91% dos pacientes portadores de EIA apresentaram baixo nível de vitamina D. A média do IMC foi de 19,6 kg/m2 para os controles e 20,3 kg/m2 no grupo 2. A análise estatística demonstrou diferença relevante (p l 0,0001) entre os níveis de vitamina D. A média e os valores mínimos de vitamina D foram respectivamente 27 e 13,6 ng/ml para o grupo 1 e 18,8 e 3,13 ng/ml para o grupo EIA. Conclusões: Pacientes portadores de EIA tinham níveis de 25-OHD mais baixos do que os pacientes saudáveis, com significância estatística. O presente estudo sugere que a deficiência da vitamina D possa estar relacionada com a etiologia da escoliose idiopática do adolescente, corroborando dados da literatura. Mais pesquisas devem ser desenvolvidas para investigar o impacto real dos níveis séricos de vitamina D sobre a fisiopatologia da EIA.

Modifiable factors of vitamin D status among a Brazilian osteoporotic population attended a public outpatient clinic

Objectives To evaluate the serum 25-hydroxyvitamin D [25(OH)D] concentration in Brazilian osteoporotic patients and the modifiable factors of vitamin D status in this population. Subjects and methods In a cross-sectional study, 363 community-dwelling patients who sought specialized medical care were evaluated between autumn and spring in São Paulo, Brazil. Serum levels of 25(OH)D and parathormone (PTH), biochemical and anthropometric measurements, and bone density scans were obtained. The group was assessed using two questionnaires: one questionnaire covered lifestyle and dietary habits, skin phototype, sun exposure, medical conditions, and levels of vitamin D supplementation (cholecalciferol); the other questionnaire assessed health-related quality-of-life. Logistic regression and a decision tree were used to assess the association between the variables and the adequacy of vitamin D status. Results The mean age of the overall sample was 67.9 ± 8.6 years, and the mean 25(OH)D concentration was 24.8 ng/mL. The prevalence of inadequate vitamin D status was high (73.3%), although 81.5% of the subjects were receiving cholecalciferol (mean dose of 8,169 IU/week). 25(OH)D was positively correlated with femoral neck bone mineral density and negatively correlated with PTH. In the multivariate analysis, the dose of cholecalciferol, engagement in physical activity and the month of the year (September) were associated with improvement in vitamin D status. Conclusions In this osteoporotic population, vitamin D supplementation of 7,000 IU/week is not enough to reach the desired 25(OH)D concentration (≥ 30 ng/mL). Engagement in physical activity and the month of the year are modifiable factors of the vitamin D status in this population.

Desenvolvimento de um radioimunoensaio de dexametasona no soro como subsídio para o diagnóstico da síndrome de Cushing

INTRODUCTION: The 1 mg dexamethasone suppression test (DxST) is widely used to screen Cushings syndrome (CS) due to its high diagnostic accuracy. CS is an endocrine-metabolic disorder caused by hypercorticism, which is characterized by the absence of cortisol suppression in DxST. OBJECTIVE: To develop a radioimmunoassay (RIA) for the measurement of serum dexamethasone (Dx) to complement DxST. METHODS: Three rabbits were inoculated with dexamethasone-21-hemisuccinate-BSA in order to choose the best antibody. Serum Dx RIA was performed according to RE 899/2003 (Agencia Nacional de Vigilância Sanitaria [ANVISA]) regulations. Serum samples from 96 volunteers from Universidade Federal de Sao Paulo (UNIFESP) and Santa Casa de Misericordia de Sao Paulo were analyzed, 67 of which were submitted to DxST and 29 were not. There were 12 patients with CS. RESULTS: The Dx antibody chosen showed good specificity. Intra- and interassay CV were < 20% with 93.8% accuracy and the lowest detection limit was 19.5 ng/dl. Serum Dx concentration was similar among both volunteers and CS patients (absence of cortisol suppression): 205 to 703 ng/dl and 174 to 661 ng/dl (95% CI), respectively. Values were undetectable among those that were not submitted to the test. Discussion: The anti-Dx antibody shows high specificity and reliability to quantify serum Dx in DxST. The Dx RIA presented reproducibility and reliability in the determination of serum Dx levels during DxST. CONCLUSION: The current RIA for serum Dx is accurate and reliable, which permits to establish a reference value range to substantiate DxST interpretation.

Vitamin D deficiency and its Relation with Adolescent Idiopathic Scoliosis

Introduction Adolescent idiopathic scoliosis (AIS) is a three-dimensional deformity of the spine occurring in up to 4% of children between 10 and 18 years of age. AIS is considered multifactorial, and probably genetics play an important role in its genesis. Family history may present several individuals affected. Classical progression risk factors such as curve magnitude, remaining growth potential, and skeletal maturity help us to identify which curves are likely to worsen. Scientists have tried to link the AIS to vitamin D receptor gene (VDRG) polymorphism. Approximately 90% of the daily recommended intake of vitamin D should be provided by sunlight through a process housed in the skin, the 10% remaining (around 400 IU) must be ingested. After hydroxylation processes hosted by the liver and kidneys, calcitriol (1,25OHVitD—active form) will play an important role in bone homeostasis and muscle strength. Vitamin D levels are considered insufficient under 30 ng/mL and deficient when lower than 20 ng/mL. Recently, bone mineral density has been studied and included as a potential explanation to the developing of skeletal deformities and its progression. Benefits of vitamin D include adequate management of muscle and bone quality, and this hormone could be implicated on AIS development. This study had the objective of comparing vitamin D serum levels of children being treated for AIS in our institution to those of normal individuals. Patients and Methods Patient recruitment was undertaken along 2013 from the outpatient clinics of Spine Pathology of the Department of Orthopedics. Children younger than 10 or older than 18 years, and those carrying neurologic or muscular disorders, congenital deformities, or genetic syndromes were excluded. Calcium, phosphorus, creatine, urea, and human parathyroid hormone dosage were taken to rule out renal or parathyroid dysfunctions. Patients were measured and weighted, and 25-OHVitD levels were determined through electrochemiluminescence by an automated test after an 8-hour fasting, during Brazilian Spring. The results were compared with a paired group composed of healthy individuals. Results The majority of the patients in both groups are nonblack females. In group 1 (controls), all children were eutrophic according to their body mass index and 63.3% showed abnormal vitamin D levels. Group 2 presented 12.7% incidence of malnutrition, and only 9% presented normal vitamin D level. Statistical analysis through unpaired t tests found relevant difference (p < 0.001) between the means of vitamin D levels of the two groups. Conclusions There is a significant difference of vitamin D levels between patients with AIS and normal individuals. However, more studies are necessary to establish a cause–consequence relation between low vitamin D levels and AIS.