Fibroblast Growth Factor 23 Is an Independent and Specific Predictor of Mortality in Patients With Heart Failure and Reduced Ejection Fraction

BACKGROUND Strategies to improve risk prediction are of major importance in patients with heart failure (HF). Fibroblast growth factor 23 (FGF-23) is an endocrine regulator of phosphate and vitamin D homeostasis associated with an increased cardiovascular risk. We aimed to assess the prognostic effect of FGF-23 on mortality in HF patients with a particular focus on differences between patients with HF with preserved ejection fraction and patients with HF with reduced ejection fraction (HFrEF). METHODS AND RESULTS FGF-23 levels were measured in 980 patients with HF enrolled in the Ludwigshafen Risk and Cardiovascular Health (LURIC) study including 511 patients with HFrEF and 469 patients with HF with preserved ejection fraction and a median follow-up time of 8.6 years. FGF-23 was additionally measured in a second cohort comprising 320 patients with advanced HFrEF. FGF-23 was independently associated with mortality with an adjusted hazard ratio per 1-SD increase of 1.30 (95% confidence interval, 1.14-1.48; P<0.001) in patients with HFrEF, whereas no such association was found in patients with HF with preserved ejection fraction (for interaction, P=0.043). External validation confirmed the significant association with mortality with an adjusted hazard ratio per 1 SD of 1.23 (95% confidence interval, 1.02-1.60; P=0.027). FGF-23 demonstrated an increased discriminatory power for mortality in addition to N-terminal pro-B-type natriuretic peptide (C-statistic: 0.59 versus 0.63) and an improvement in net reclassification index (39.6%; P<0.001). CONCLUSIONS FGF-23 is independently associated with an increased risk of mortality in patients with HFrEF but not in those with HF with preserved ejection fraction, suggesting a different pathophysiologic role for both entities.

Bone metabolic and hormonal characteristics of Australian Antarctic expeditioners

Summary: Serum 25(OH)D levels decline without sunlight exposure. We studied 120 expeditioners to Antarctica to determine the skeletal and hormonal responses to sunlight deprivation. With emerging vitamin D insufficiency, serum calcium decreased, PTH increased, and bone loss at the proximal femur was observed. Baseline serum 25(OH)D levels >100 nmol/L prevented vitamin D insufficiency. Introduction: Vitamin D stores deplete without adequate sunlight exposure unless supplementation is provided. We studied 120 healthy adults who spent a year in Antarctica as a model for sunlight deprivation to define the timing and magnitude of the skeletal and hormonal responses to emerging vitamin D insufficiency. Methods: Fasting blood samples were assessed at baseline, 6 and 12 months for serum 25-hydroxyvitamin D (25(OH)D), osteocalcin (OC), bone formation (P1NP) and resorption (CTx), PTH and calcium. Lumbar spine and proximal femur BMD was measured using DXA. Differences over time were determined using repeated measures ANOVA. Percent changes were expressed as (Delta value/(value A +value B)/2)x100. Relationships between outcome measures were determined using Spearman's correlations. Results: Vitamin D insufficiency (<50 nmol/L) was observed in 85% of expeditioners by 6 months when serum calcium decreased and PTH increased (p<0.01). By 12 months, OC increased by 7.4±3.0% (p<0.05), and BMD decreased by 1.0±2.0% at the total proximal femur (p<0.05). For those with vitamin D sufficiency at baseline (>50 nmol/L), sunlight deprivation produced vitamin D insufficiency within 4 months unless baseline values were >100 nmol/L. Conclusion: Supplementation may be necessary for expeditioners with limited access to UV light.

Concentrations of PCBs, DDT and thyroid hormones in gray (Halichoerus grypus) and ringed (Phoca hispida) seals

The high levels of polychlorinated biphenyls (PCBs) and DDT in gray seal (Halichoerus grypus) and ringed seal (Phoca hispida botnica) in the Baltic Sea have been associated with pathological disruptions, including bone lesions and reproductive failures. The underlying environmental and toxicological mechanisms leading to these pathological changes are not yet fully understood. The present study investigated the relationship between the individual contaminant load and bone- and thyroid-related effects in adult gray seals (n = 30) and ringed seals (n = 46) in the highly contaminated Baltic Sea and in reference areas (Sable Island, Canada, and Svalbard, Norway). In the gray seals, multivariate and correlation analyses revealed a clear relationship between circulating 1,25-dihydroxyvitamin D3 (1,25(OH)2D), calcium, phosphate, and thyroid hormone (TH) levels and hepatic PCB and DDT load, which suggests contaminant-mediated disruption of the bone and thyroid homeostasis. Contaminants may depress 1,25(OH)2D levels or lead to hyperthyroidism, which may cause bone resorption. In the ringed seals, associations between circulating 1,25(OH)2D, THs, and hepatic contaminants were less prominent. These results suggest that bone lesions observed in the Baltic gray seals may be associated with contaminant-mediated vitamin D and thyroid disruption.

Cloning and expression of rat 25-hydroxyvitamin D3-1α-hydroxylase cDNA

A full-length cDNA for the rat kidney mitochondrial cytochrome P450 mixed function oxidase, 25-hydroxyvitamin D3-1α-hydroxylase (P4501α), was cloned from a vitamin D-deficient rat kidney cDNA library and subcloned into the mammalian expression vector pcDNA 3.1(+). When P4501α cDNA was transfected into COS-7 transformed monkey kidney cells, they expressed 25-hydroxyvitamin D3-1α-hydroxylase activity. The sequence analysis showed that P4501α was of 2,469 bp long and contained an ORF encoding 501 amino acids. The deduced amino acid sequence showed a 53% similarity and 44% identity to the vitamin D3-25-hydroxylase (CYP27), whereas it has 42.6% similarity and 34% identity with the 25-hydroxyvitamin D3-24-hydroxylase (CYP24). Thus, it composes a new subfamily of the CYP27 family. Further, it is more closely related to the CYP27 than to the CYP24. The expression of P4501α mRNA was greatly increased in the kidney of vitamin D-deficient rats. In rats with the enhanced renal production of 1α,25-dihydroxyvitamin D3 (rats fed a low Ca diet), P4501α mRNA was greatly increased in the renal proximal convoluted tubules.

Complete structure of the human alpha-albumin gene, a new member of the serum albumin multigene family.

The nucleotide sequence of the human alpha-albumin gene, including 887 bp of the 5'-flanking region and 1311 bp of the 3-flanking region (24,454 in total), was determined from three overlapping lambda phage clones. The sequence spans 22,256 bp from the cap site to the polyadenylylation site, revealing a gene structure of 15 exons separated by 14 introns. The methionine initiation codon ATG is within exon 1; the termination codon TGA is within exon 14. Exon 15 is entirely untranslated and contains the polyadenylylation signal AATAAA. The deduced polypeptide chain is composed of a 21-amino-acid leader peptide, followed by 578 amino acids of the mature protein. There are seven repetitive DNA elements (Alu and Kpn) in the introns and 3-flanking region. The sizes of the 15 alpha-albumin exons match closely those of the albumin, alpha-fetoprotein, and vitamin D-binding protein genes. The exons are symmetrically placed within the three domains of the individual proteins, and they share a characteristic codon splitting pattern that is conserved among members of the gene family. The results provide strong evidence that alpha-albumin belongs to, and most likely completes with, the serum albumin gene family. Based on structural similarity, alpha-albumin appears to be most closely related to alpha-fetoprotein. The complete structure of this family of four tandemly linked genes provides a well-characterized approximately 200 kb locus in the 4q subcentromeric region of the human genome.

1,25-Dihydroxyvitamin D3 reversibly blocks the progression of relapsing encephalomyelitis, a model of multiple sclerosis.

Experimental autoimmune encephalomyelitis (EAE) is an autoimmune disease believed to be a model for the human disease multiple sclerosis (MS). Induced by immunizing B10.PL mice with myelin basic protein (MBP), EAE was completely prevented by the administration of 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3]. 1,25-(OH)2D3 could also prevent the progression of EAE when administered at the appearance of the first disability symptoms. Withdrawal of 1,25-(OH)2D3 resulted in a resumption of the progression of EAE. Thus, the block by 1,25-(OH)2D3 is reversible. A deficiency of vitamin D resulted in an increased susceptibility to EAE. Thus, 1,25-(OH)2D3 or its analogs are potentially important for treatment of MS.

A functional Rev-erb alpha responsive element located in the human Rev-erb alpha promoter mediates a repressing activity.

Rev-erb alpha belongs to the nuclear receptor superfamily, which contains receptors for steroids, thyroid hormones, retinoic acid, and vitamin D, as well as "orphan" receptors. No ligand has been found for Rev-erb alpha to date, making it one of these orphan receptors. Similar to some other orphan receptors, Rev-erb alpha has been shown to bind DNA as a monomer on a specific sequence called a Rev-erb alpah responsive element (RevRE), but its transcriptional activity remains unclear. In this paper, we characterize a functional RevRE located in the human Rev-erb alpha promoter itself. We also present evidence that (i) Rev-erb alpha mediates transcriptional repression of its own promoter in vitro, (ii) this repressing effect strictly depends on the binding of Rev-erb alpha to its responsive element and is transferable to a heterologous promoter; and (iii) Rev-erb alpha binds to this responsive sequence as a homodimer.

Concentrações séricas de 25 (OH) e perfil metabólico mediados pela adiposidade

Introdução: Baixas concentrações séricas de hidroxivitamina D (25[OH]D) e o excesso de peso atingiram níveis epidêmicos em todo o mundo. Estudos relatam que concentrações séricas de vitamina D estão associadas às alterações lipídicas, glicolíticas e inflamatórias; e estas alterações são conhecidamente mediadas pela adiposidade. Dessa forma, a vitamina D pode atuar de forma benéfica sobre o perfil metabólico em adolescentes, adultos e idosos. Objetivo: Investigar e descrever as associações entre as concentrações séricas de 25(OH)D e o perfil metabólico, mediadas pela adiposidade em adolescentes, adultos e idosos. Metodologia: Inicialmente, foi utilizada subamostra do Inquérito de Saúde de São Paulo (ISA-Capital), estudo transversal, de base populacional (n=281), para investigar a associação entre as concentrações séricas de vitamina D e marcadores inflamatórios em adultos brasileiros. Posteriormente, foram utilizados dados do estudo Healthy Lifestyle in Europe by Nutrition in Adolescents-(HELENA), estudo multicêntrico transversal da população de adolescentes européia, com o intuito de avaliar as alterações nos marcadores lipídicos e de homeostase da glicose mediados pela deficiência de vitamina D e obesidade. Finalmente, foi analisada a amostra do estudo PHYSMED, um estudo transversal com idosos não institucionalizados para verificar associações entre concentrações séricas de vitamina D, perfil lipídico e composição corporal em idosos espanhóis aparentemente saudáveis. Resultados: Nos adultos, observou-se uma associação negativa entre as concentrações de TNF-alfa e de IL-6 e as concentrações séricas de 25(OH)D em indivíduos com peso normal. Nos adolescentes, as concentrações de 25(OH)D foram associadas de forma independente e positiva com o Quantitative Insulin Sensitivity Check Index-QUICKI (p <0.001) e negativamente associada com o IMC (p <0.05). Também foi observado que o aumento do IMC esteve associado com um aumento de 1.93 vezes maior chance de deficiência de vitamina D (IC de 95 por cento = 1.03 - 3.62; p = 0.040). Em idosos, verificou-se que as concentrações séricas de 25(OH)D foram associadas com o IMC (p = 0.04), a circunferência da cintura (p = 0.004), CT/HDL-c (p = 0.026) e o HDL-c (p = 0.001). Adicionalmente, foi observado que idosos com concentrações de HDL-c <40mg/dl possuíam 1.7 vezes maior chance de apresentarem deficiência de vitamina D em comparação com aqueles que possuíam concentrações de HDL-c >40 mg/dl (95 por cento IC = 1.10 a 2.85; p = 0.017) e o aumento na circunferência da cintura também foi associado com um maior risco de deficiência de vitamina D (95 por cento IC =0.96-1.00; p = 0.04). Conclusão: A composição corporal interage com as concentrações de 25(OH)D modulando a resposta inflamatória, à homeostase da glicose e também o perfil lipídico. Indivíduos sem deficiência de vitamina D apresentam melhor perfil metabólico e também melhor composição, sugerindo que a suficiência de vitamina D pode ter um papel importante nas condições metabólicas mediadas pela adiposidade.

Efeito do treinamento físico associado a um programa de perda de peso no controle clínico e nos fatores de saúde relacionados à qualidade de vida em asmáticos obesos

Introdução: O fenótipo asma-obeso caracteriza-se por uma asma mais grave, no qual o controle clínico é mais difícil de ser alcançado, mesmo sob terapia medicamentosa otimizada. A cirurgia bariátrica tem sido recomendada para perda de peso e melhora dos sintomas, porém os benefícios de intervenções não-cirúrgicas têm sido pouco estudados. Objetivo: Avaliar o efeito do treinamento físico associado a um programa de perda de peso no controle clínico da asma, qualidade de vida e sintomas psicossociais em asmáticos obesos. Métodos: 55 pacientes com asma moderada a grave e obesidade grau II (IMC >= 35 e < 39.9 kg/m2) foram alocados em 2 grupos: programa de perda de peso + placebo (PP+P) ou programa de perda de peso + exercícios (PP+E), sendo que o programa de perda de peso incluiu terapia nutricional e psicológica (12 sessões semanais de 60 minutos cada). O grupo PP+E associou exercícios aeróbios e resistidos programa de perda de peso, enquanto o grupo PP+P associou exercícios placebo (respiratórios e alongamentos), 2xvezes/semana, 60 minutos/sessão durante 3 meses. Antes e após as intervenções, foram avaliados o controle clínico da asma, os fatores de saúde relacionados a qualidade de vida (FSRQV), a capacidade física, a composição corporal, os sintomas de ansiedade e depressão, a qualidade do sono, a função pulmonar e as inflamações das vias aéreas e sistêmica. A comparação dos dados contínuos entre os grupos foi realizada por ANOVA de dois fatores com medidas repetidas e das variáveis categóricas pelo teste qui-quadrado. A correlação linear e a regressão linear múltipla foram utilizadas para avaliar associações entre as variáveis avaliadas. Resultados: Foram analisados os resultados de 51 pacientes que foram reavaliados. Comparado com o grupo PP+P, os pacientes que realizaram exercício apresentaram melhora no controle clínico da asma (- 0,7 [-1,3 - -0,3] vs. -0,3 [-0,9 - 0,4] escore ACQ; p=0,01) e nos FSRQV (0,8 [0,3 -2,0] vs. 0,4 [-0,3 - 0,9] escore AQLQ; p=0,02), respectivamente. Essa melhora parece ter sido consequência do aumento do condicionamento físico (3,0 [2,4-4,0] vs. 0,9 [-0,3-1,3] mL.O2/Kg/min; p < 0,001) e da perda de peso (6,8±3,5% vs. 3,1±2,6% do peso corpóreo; p < 0,001) nos pacientes do grupo PP+E, que também apresentaram uma melhora dos sintomas de depressão, da qualidade do sono (ronco, latência e eficiência) e dos níveis séricos de vitamina D. Houve também melhora da função pulmonar (capacidade vital forçada e volume de reserva expiratório) e das inflamações das vias aéreas (FeNO) e sistêmica (CCL2, CXCL9, IL-4, IL-6, TNF-alfa, IL-10 e leptina/adiponectina), que parecem ser possíveis mecanismos associados à melhora do controle clinico da asma nos pacientes do grupo PP+E (p < 0,05 para todas variáveis apresentadas). Conclusão: A inclusão do treinamento físico em um programa de perda de peso a curto prazo deve ser considerada como uma intervenção eficiente para associar à terapia medicamentosa da asma na melhora do controle clínico em asmáticos obesos

Hábitos alimentares durante a gravidez : identificação dos hábitos alimentares e fontes de informação consultadas pelas utentes da consulta pré-natal do Hospital de Santa Maria

Trabalho Final do Curso de Mestrado Integrado em Medicina, Faculdade de Medicina, Universidade de Lisboa, 2014

Prostaglandins are important in thermoregulation of a reptile (Pogona vitticeps)

The effectiveness of behavioural thermoregulation in reptiles is amplified by cardiovascular responses, particularly by differential rates of heart beat in response to heating and cooling (heart-rate hysteresis). Heart-rate hysteresis is ecologically important in most lineages of ectothermic reptile' and we demonstrate that heart-rate hysteresis in the lizard Pogona vitticeps is mediated by prostaglandins. In a control treatment (administration of saline), heart rates during heating were significantly faster than during cooling at any given body temperature. When cyclooxygenase 1 and 2 enzymes were inhibited, heart rates during heating were not significantly different from those during cooling. Administration of agonists showed that thromboxane B-2 did not have a significant effect on heart rate, but prostacyclin and prostaglandin F-2alpha caused a significant increase (3.5 and 13.6 beats min(-1), respectively) in heart rate compared with control treatments. We speculate that heart-rate hysteresis evolved as a thermoregulatory mechanism that may ultimately be controlled by neurally induced stimulation of nitric oxide production, or maybe via photolytically induced production of vitamin D.

Characterization of the human sulfate anion transporter (hsat-1) protein and gene (SAT1; SLC26A1)

Sulfate plays an essential role during growth, development, bone/cartilage formation, and cellular metabolism. In this study, we have isolated the human sulfate anion transporter cDNA (hsat-1; SCL26A1) and gene (SAT1), determined its protein function in Xenopus oocytes and characterized SAT1 promoter activity in mammalian renal cell lines. hsat-1 encodes a protein of 75 kDa, with 12 putative transmembrane domains, that induces sulfate, chloride, and oxalate transport in Xenopus oocytes. hsat-1 mRNA is expressed most abundantly in the kidney and liver, with lower levels in the pancreas, testis, brain, small intestine, colon, and lung. The SAT1 gene is comprised of four exons stretching 6 kb in length, with an alternative splice site formed from an optional exon. SAT1 5' flanking region led to promoter activity in renal OK and LLC-PK1 cells. Using SAT1 5' flanking region truncations, the first 135 bp was shown to be sufficient for basal promoter activity. Mutation of the activator protein-1 (AP-1) site at position 252 in the SAT1 promoter led to loss of transcriptional activity, suggesting its requirement for SAT1 basal expression. This study represents the first functional characterization of the human SAT1 gene and protein encoded by the anion transporter hsat-1.

The mouse sulfate anion transporter gene Sat1 (Slc26a1): Cloning, tissue distribution, gene structure, functional characterization, and transcriptional regulation by thyroid hormone

Sulfate (SO42-) is required for bone/cartilage formation and cellular metabolism. sat-1 is a SO42- anion transporter expressed on basolateral membranes of renal proximal tubules, and is suggested to play an important role in maintaining SO42- homeostasis. As a first step towards studying its tissue-specific expression, hormonal regulation, and in preparation for the generation of knockout mice, we have cloned and characterized the mouse sat-1 cDNA (msat-1), gene (sat1; Slc26a1) and promoter region. msat-1 encodes a 704 amino acid protein (75.4 kDa) with 12 putative transmembrane domains that induce SO42- (also oxalate and chloride) transport in Xenopus oocytes. msat-1 mRNA was expressed in kidney, liver, cecum, calvaria, brain, heart, and skeletal muscle. Two distinct transcripts were expressed in kidney and liver due to alternative utilization of the first intron, corresponding to an internal portion of the 5'-untranslated region. The Sa1 gene (similar to6 kb) consists of 4 exons. Its promoter is similar to52% G+C rich and contains a number of well-characterized cis-acting elements, including sequences resembling hormone responsive elements T3REs and VDREs. We demonstrate that Sat1 promoter driven basal transcription in OK cells was stimulated by tri-iodothyronine. Site-directed mutagenesis identified an imperfect T3RE at -454-bp in the Sat1 promoter to be responsible for this activity. This study represents the first characterization of the structure and regulation of the Sat1 gene encoding a SO42-/chloride/oxalate anion transporter.

Quantitative Association Tests of Immune Responses to Antigens of Mycobacterium Tuberculosis: A Study of Twins in West Africa

There is now considerable evidence that host genetic factors are important in determining the outcome of infection with Mycobacterium tuberculosis (MTB). The aim of this study was to assess the role of several candidate genes in the variation observed in the immune responses to MTB antigens. In-vitro assays of T-cell proliferation, an in-vivo intradermal delayed hypersensitivity response; cytokine and antibody secretions to several mycobacterial peptide antigens were assessed in healthy, but exposed, West African twins. Candidate gene polymorphisms were typed in the NRAMP1, Vitamin D receptor, IL10, IL4, IL4 receptor and CTLA-4 genes. Variants of the loci IL10 (-1082 G/A), CTLA-4 (49 A/G) and the IL4 receptor (128 A/G) showed significant associations with immune responses to several antigens. T-cell proliferative responses and antibody responses were reduced, TNF-alpha responses were increased for subjects with the CTLA-4 G allele. The T-cell proliferative responses of subjects with IL10 GA and GG genotypes differed significantly. IL4 receptor AG and GG genotypes also showed significant differences in their T-cell proliferative responses to MTB antigens. These results yield a greater understanding of the genetic mechanisms that underlie the immune responses in tuberculosis and have implications for the design of therapeutic interventions.