Adult granulosa cell tumors of the ovary: a retrospective study of 30 cases with respect to the expression of steroid synthesis enzymes / 부인종양

OBJECTIVE: Some, but not all, granulosa cell tumors are characterized by estrogen production. This study was designed to determine whether there are clinical or pathological variations in granulosa cell tumors in relation to the expression of sex steroid synthesis enzymes. METHODS: Clinical symptoms, serum hormonal values, and histology of 30 granulosa cell tumor patients who underwent surgery between 2002 and 2014 were retrospectively reviewed. RESULTS: Most patients presented with abnormal genital bleeding including abnormal menstrual cycles. Eight of 16 patients older than 50 years had endometrial hyperplasia and one had endometrial cancer. Serum 17β-estradiol (E2) levels tended to be higher in patients over 50 years of age (p=0.081). Serum follicle-stimulating hormone (FSH) levels were low in all patients irrespective of serum E2 levels. Magnetic resonance imaging revealed a thicker endometrium in older as compared to younger patients (p<0.05). Tumor cells in the majority of cases were positive for inhibin α and P450 aromatase, irrespective of age and serum E2 levels. P450 17α-hydroxylase (P450c17) expression varied among cases. P450c17 was strongly positive in luteinized tumor cells and weakly positive in theca cells and fibroblasts. High E2 levels were associated with P450c17-positive cells in the tumor (p<0.05). CONCLUSION: The expression of hormone-synthesizing enzymes divides granulosa cell tumors into 2 distinct types; tumors with P450c17-positive cells show elevated serum E2 and related clinical symptoms, while tumors without these cells show symptoms related to FSH suppression by inhibin.

Maternal protein restriction during lactation modulated the expression and activity of rat offspring hepatic CYP1A1, CYP1A2, CYP2B1, CYP2B2, and CYP2E1 during development

Early nutrition plays a long-term role in the predisposition to chronic diseases and influences the metabolism of several drugs. This may happen through cytochromes P450 (CYPs) regulation, which are the main enzymes responsible for the metabolism of xenobiotics. Here, we analyzed the effects of maternal protein restriction (MPR) on the expression and activity of hepatic offspring’s CYPs during 90 days after birth, using Wistar rats as a mammal model. Hepatic CYP1A1, CYP1A2, CYP2B1, CYP2B2 and CYP2E1 mRNA and protein expression, and associated catalytic activities (ECOD, EROD, MROD, BROD, PROD and PNPH) were evaluated in 15-, 30-, 60-, and 90-day-old offspring from dams fed with either a 0% protein (MPR groups) or a standard diet (C groups) during the 10 first days of lactation. Results showed that most CYP genes were induced in 60- and 90-day-old MPR offspring. The inductions detected in MPR60 and MPR90 were of 5.0- and 2.0-fold (CYP1A2), 3.7- and 2.0-fold (CYP2B2) and 9.8- and 5.8– fold (CYP2E1), respectively, and a 3.8-fold increase of CYP2B1 in MPR90. No major alterations were detected in CYP protein expression. The most relevant CYP catalytic activities’ alterations were observed in EROD, BROD and PNPH. Nevertheless, they did not follow the same pattern observed for mRNA expression, except for an induction of EROD in MPR90 (3.5-fold) and of PNPH in MPR60 (2.2-fold). Together, these results suggest that MPR during lactation was capable of altering the expression and activity of the hepatic CYP enzymes evaluated in the offspring along development.

SNP genetic polymorphisms of MDR-1, CYP1A2 and CYPB11 genes in four canine breeds upon toxicological evaluation

The fields of pharmacogenetics and pharmacogenomics have become increasingly promising regarding the clinical application of genetic data to aid in prevention of adverse reactions. Specific screening tests can predict which animals express modified proteins or genetic sequences responsible for adverse effects associated with a drug. Among the genetic variations that have been investigated in dogs, the multidrug resistance gene (MDR) is the best studied. However, other genes such as CYP1A2 and CYP2B11 control the protein syntheses involved in the metabolism of many drugs. In the present study, the MDR-1, CYP1A2 and CYP2B11 genes were examined to identify SNP polymorphisms associated with these genes in the following four canine breeds: Uruguayan Cimarron, Border Collie, Labrador Retriever and German Shepherd. The results revealed that several SNPs of the CYP1A2 and CYP2B11 genes are potential targets for drug sensitivity investigations.

Interdisciplinarity and nursing research: opportunities and challenges / Interdisciplinaridade e pesquisa em enfermagem: oportunidades e desafios / La interdisciplinariedad y la investigación en enfermería: oportunidades y desafíos

Interdisciplinary collaboration is widely recognized and considered essential for optimizing the development of knowledge and practice. However, interdisciplinarity is commonly accepted as an unquestioned good; rarely examined as both a source of benefit as well as difficulty for nursing and other disciplines. The aim of this article is to critically examine the opportunities and challenges that interdisciplinarity can provide for research in nursing and other disciplines. Based on a North American perspective, I describe the emergence of uni-disciplinary nursing research and the knowledge exchanges that occurred between nursing and other disciplines. I discuss the rise of interdisciplinary research, outline several examples of nursing participation in interdisciplinarity, and highlight the prominent benefits and difficulties associated with interdisciplinary research. I argue that authentic collaboration is required to conduct meaningful interdisciplinary research and describe how this can be promoted.

Colaboração interdisciplinar é amplamente reconhecida e considerada essencial para a otimização do desenvolvimento do conhecimento e prática. No entanto, a interdisciplinaridade é comumente aceita como um bem inquestionável, raramente examinado tanto como uma fonte de benefícios, bem como dificuldade para a enfermagem e outras disciplinas. O objetivo deste artigo é analisar criticamente as oportunidades e desafios que a interdisciplinaridade pode oferecer para a pesquisa em enfermagem e outras disciplinas. Com base em uma perspectiva norte-americana, descreve-se o surgimento de pesquisas em enfermagem unidisciplinar e as trocas de conhecimento que ocorreram entre a enfermagem e outras disciplinas. Discute-se a ascensão da pesquisa interdisciplinar, delineiam-se vários exemplos de participação da enfermagem na interdisciplinaridade, e destacam-se os benefícios proeminentes e dificuldades associadas com a pesquisa interdisciplinar. Defende-se que a colaboração autêntica é necessária para conduzir a pesquisa interdisciplinar significativa e descreve-se como isso pode ser promovido.

La colaboración interdisciplinaria es ampliamente reconocida y considerada esencial para optimizar el desarrollo del conocimiento y la práctica. Sin embargo, la interdisciplinariedad es comúnmente aceptada como un bien incuestionable; rara vez examinada tanto como una fuente de beneficio, así como de dificultad para la enfermería y otras disciplinas. El objetivo de este artículo es examinar críticamente las oportunidades y desafíos que la interdisciplinariedad puede proporcionar para la investigación en enfermería y otras disciplinas. Sobre la base de una perspectiva norteamericana, describe-se el surgimiento de la investigación en enfermería unidisciplinaria y los intercambios de conocimientos que se produjeron entre la enfermería y otras disciplinas. Se discute el aumento de la investigación interdisciplinaria, esbozan-se varios ejemplos de la participación de enfermería en la interdisciplinariedad, y destacan-se los beneficios y las dificultades asociadas con la investigación interdisciplinaria. Argumenta-sé que se requiere auténtica colaboración para llevar a cabo la investigación interdisciplinaria significativa y describe-se la forma en que esto puede ser promovido. .

Insights into battles between Mycobacterium tuberculosis and macrophages

As the first line of immune defense for Mycobacterium tuberculosis (Mtb), macrophages also provide a major habitat for Mtb to reside in the host for years. The battles between Mtb and macrophages have been constant since ancient times. Triggered upon Mtb infection, multiple cellular pathways in macrophages are activated to initiate a tailored immune response toward the invading pathogen and regulate the cellular fates of the host as well. Toll-like receptors (TLRs) expressed on macrophages can recognize pathogen-associated-molecular patterns (PAMPs) on Mtb and mediate the production of immune-regulatory cytokines such as tumor necrosis factor (TNF) and type I Interferons (IFNs). In addition, Vitamin D receptor (VDR) and Vitamin D-1-hydroxylase are up-regulated in Mtb-infected macrophages, by which Vitamin D participates in innate immune responses. The signaling pathways that involve TNF, type I IFNs and Vitamin D are inter-connected, which play critical roles in the regulation of necroptosis, apoptosis, and autophagy of the infected macrophages. This review article summarizes current knowledge about the interactions between Mtb and macrophages, focusing on cellular fates of the Mtb-infected macrophages and the regulatory molecules and cellular pathways involved in those processes.

New insights into steroidogenesis in normo- and hyperandrogenic polycystic ovary syndrome patients / Novos aspectos na esteroidogênese em pacientes normo ou hiperandrogênicas com síndrome de ovários policísticos

OBJECTIVE: This study sought to examine corticosteroidogenic enzyme activities in normo- and hyperandrogenic polycystic ovary syndrome (PCOS) patients. SUBJECTS AND METHODS: This cohort study included 81 patients with biochemical hyperandrogenism and 41 patients with normal androgen levels. Enzyme activities were assessed according to the serum steroid product/precursor ratios at baseline and after adrenal stimulation. RESULTS: At baseline, in the delta 4 (Δ4) pathway, hyperandrogenic patients showed greater 17-hydroxylase and 17,20 lyase activities in converting progesterone (P4) into 17-hydroxyprogesterone (17-OHP4) and 17-hydroxypregnenolone (17-OHPE) into androstenedione (A) (p = 0.0005 and p = 0.047, respectively) compared to normoandrogenic patients. In the delta 5 (Δ5) pathway, the 17-hydroxylase and 17,20 lyase enzymes showed similar activities in both groups. Hyperandrogenic patients presented lower 21-hydroxylase, lower 11β-hydroxylase (p = 0.0001), and statistically significant increases in 3β-hydroxysteroid dehydrogenase II (3β-HSDII) activities (p < 0.0001). Following tetracosactrin stimulation, only the 17,20 lyase activity remained up-regulated in the Δ4 pathway (p < 0.0001). CONCLUSION: Hyperandrogenic patients had higher 17,20 lyase activity, both at baseline and after adrenal stimulation. Greater conversion of dehydroepiandrosterone (DHEA) into A with normal conversion of 17-OHPE to 17-OHP4 in hyperandrogenic PCOS patients indicated different levels of 3β-HSDII activity in adrenal cells, and hyperandrogenic patients had lower 11β-hydroxylase and 21-hydroxylase activities.

OBJETIVO: O objetivo deste estudo foi examinar a atividade de enzimas responsáveis pela produção de corticosteroides em pacientes normo e hiperandrogênicas com síndrome de ovários policísticos (SOP). SUJEITOS E MÉTODOS: A coorte estudada incluiu 81 pacientes com hiperandrogenismo bioquímico e 41 pacientes com níveis normais de androgênio. A atividade enzimática foi avaliada de acordo com as proporções de produto/precursor do esteroide sérico, no momento inicial do estudo e depois de estimulação adrenal. RESULTADOS: No momento inicial, na via delta 4 (Δ4), as pacientes hiperandrogênicas mostraram maior atividade da 17-hidroxilase e 17,20 liase na conversão da progesterona (P4) em 17-hidroxiprogesterona (17-OHP4) e na conversão da 17-hidroxipregnenolona (17-OHPE) em androstenediona (A) (p = 0,0005 e p = 0,047, respectivamente) em comparação com pacientes normoandrogênicas. Na via delta 5 (Δ5), a 17-hidroxilase e a 17,20 liase mostraram atividades similares nos dois grupos. As pacientes hiperandrogênicas mostraram menor atividade da 21-hidroxilase, menor atividade da 11β-hidroxilase (p = 0,0001) e aumento estatisticamente significativo na atividade da 3β-hidroxiesteroide desidrogenase II (3β-HSDII) (p < 0.0001). Após a estimulação com tetracosactrin, apenas a atividade da 17,20 liase permaneceu regulada para cima na via Δ4 (p < 0.0001). CONCLUSÃO: As pacientes hiperandrogênicas apresentaram atividade mais alta da 17,20 liase, tanto no momento inicial quanto depois da estimulação adrenal. Maior conversão da desidroepiandrosterona (DHEA) em A com conversão normal da 17-OHPE em 17-OHP4 em pacientes hiperandrogênicas com SOP indica níveis diferentes de atividade da 3β-HSDII em células da adrenal, e pacientes hiperandrogênicas apresentaram menores atividades da 11β-hidroxilase e da 21-hidroxilase.

O sistema endocrinológico vitamina D / The vitamin D endocrine system

O sistema endocrinológico vitamina D é constituído por um grupo de moléculas secosteroides derivadas do 7-deidrocolesterol, incluindo a forma ativa 1,25-diidroxi-vitamina D (1,25(OH)2D), seus precursores e metabólitos, sua proteína transportadora (DBP), seu receptor nuclear (VDR) e as enzimas do complexo do citocromo P450 envolvidas nos processos de ativação e inativação dessas moléculas. Os efeitos biológicos da 1,25(OH)2D são mediados pelo VDR, um fator de transcrição ativado por ligante, presente em quase todas as células humanas, e que pertence à família de receptores nucleares. Além dos clássicos papéis de reguladora do metabolismo do cálcio e da saúde óssea, as evidências sugerem que a 1,25(OH)2D module direta ou indiretamente cerca de 3 por cento do genoma humano, participando do controle de funções essenciais à manutenção da homeostase sistêmica, tais como crescimento, diferenciação e apoptose celular, regulação dos sistemas imunológico, cardiovascular e musculoesquelético, e no metabolismo da insulina. Pela influência crítica que esse sistema exerce em vários processos do equilíbrio metabólico sistêmico, é importante que os ensaios laboratoriais utilizados para sua avaliação apresentem alta acurácia e reprodutibilidade, permitindo que sejam estabelecidos pontos de corte que, além de serem consensualmente aceitos, expressem adequadamente o grau de reserva de vitamina D do organismo e reflitam os respectivos impactos clínico-metabólicos na saúde global do indivíduo.

The vitamin D endocrine system comprises a group of 7-dehydrocholesterol-derived secosteroid molecules, including its active metabolite 1,25-dihydroxy-vitamin D (1,25(OH)2D), its precursors and other metabolites, its binding protein (DBP) and nuclear receptor (VDR), as well as cytochrome P450 complex enzymes participating in activation and inactivation pathways of those molecules. The biologic effects of 1,25(OH)2D are mediated by VDR, a ligand-activated transcription factor which is a member of the nuclear receptors family, spread in almost all human cells. In addition to its classic role in the regulation of calcium metabolism and bone health, evidence suggests that 1,25(OH)2D directly or indirectly modulates about 3 percent of the human genome, participating in the regulation of chief functions of systemic homeostasis, such as cell growth, differentiation and apoptosis, regulation of immune, cardiovascular and musculoskeletal systems, and insulin metabolism. Given the critical influence of the vitamin D endocrine system in many processes of systemic metabolic equilibrium, the laboratory assays available for the evaluation of this system have to present high accuracy and reproducibility, enabling the establishment of cutoff points that, beyond being consensually accepted, reliably express the vitamin D status of the organism, and the respective clinical-metabolic impacts on the global health of the individual.

Effects of brucine combined with glycyrrhetinic acid or liquiritin on rat hepatic cytochrome P450 activities in vivo / 药学学报

Abstract: The activities of four CYP450 enzymes (CYP3A, 1A2, 2El and 2C) and the mRNA expression levels of CYP1A2, 2El, 2Cll and 3A1 in rat liver were determined after Wistar rats were orally administered with brucine (BR) at three dosage levels (3, 15 and 60 mg.kg-1 per day) and the high dose of BR combined with glycyrrhetinic acid (GA, 25 mg.kg-1 per day) or liquiritin (LQ, 20 mg.kg-1 per day) for 7 consecutive days. Compared with the control, brucine caused 24.5% and 34.6% decrease of CYP3A-associated testosterone 6beta-hydroxylation (6betaTesto-OH) and CYP2C-associated tolbutamide hydroxylation (Tol-OH), respectively, and 146.1% increase of CYP2El-associated para-nitrophenol hydroxylation (PNP-OH) at the high dose level. On the other hand, (BR+GA) caused 51.4% and 33.5% decrease, respectively, of CYP2El-associated PNP-OH and CYP1A2-associated ethoxyresorufin-O-de-ethylation (EROD) as compared with the high dose of BR group. Meanwhile, (BR+LQ) caused 41.1% decrease of CYP2El-associated PNP-OH and 37.7% increase of CYP2C-associated Tol-OH. The results indicated that the co-administration of BR with GA or LQ had effect on mRNA expression and activities of the CYP450 enzymes mentioned above to some extent, and the in vivo antagonism of LQ on BR-induced CYPs adverse effects and the in vivo inhibitory action of GA on CYP2E1 and 1A2 might play an important role in the detoxification of Radix Glycyrrhizae against Strychnos nux-vomica L.

The Role of MAPK Signaling of 1,25(OH)2D3-induced CYP24 Expression in Activated Human Macrophages / 대한류마티스학회지

OBJECTIVE: Several important roles of 1,25(OH)2D3 have been recognized in the immune system. The availability of 1,25(OH)2D3 at the cellular level is significantly influenced by the relative abundance of enzymes to synthesize (CYP27B1) and catabolize (CYP24) 1,25(OH)2D3. In this study, we examined the effect of 1,25(OH)2D3 on the expression of the CYP24 gene and the role of MAPK for the induction of CYP24 by 1,25(OH)2D3 in activated human macrophages. METHODS: For obtaining human activated macrophages, we treated U937 cells with PMA and we cultured these cells for 24 hours to adhere. After 24 hours treatment with PMA, the differentiated cells were washed with phosphate buffered saline (PBS), and then they were used for examining the effect of 1,25(OH)2D3 on the expression of the CYP24 gene. The mRNA expressions of the vitamin-D3 inducible genes were measured by real-time PCR, and the change of the protein expression by 1,25(OH)2D3 was measured by immunoblotting. RESULTS: 1,25(OH)2D3 significantly induced the expression of CYP24 in the U937 cells and the 1,25(OH)2D3-induced expression of CYP24 was strongly augmented in the PMA-differentiated U937 cells. The 1,25(OH)2D3-induced expression of CYP24 was mediated by Erk1/2 and p38 MAPKs. Parallel to the induced expression of CYP24, 1,25(OH)2D3 induced the expression and phosphorylation of the CCAAT enhancer-binding protein (C/EBPbeta). CONCLUSION: In this study, we found that 1,25(OH)2D3 inducedthe expression of CYP24 via activation of MAPKs. These results suggest that MAPK inhibitors may be useful for the treatment of inflammatory conditions, in which active vitamin D3 can be used as the therapeutic molecule, by increasing the availability of 1,25(OH)2D3.

A young woman with hypogonadism, hypertension and hypokalaemia

We report a case of a 16 years old girl who presented sequentially with primary amenorrhoea, hypertension and hypokalaemia. Eight years later, she was finally diagnosed with 17alpha-hydroxylase deficiency congenital adrenal hyperplasia. Previous antihypertensive medications were stopped. Hydrocortisone alone successfully maintained normotension and normokalaemia.

Mutations in the vitamin D receptor gene in four patients with hereditary 1, 25-dihydroxyvitamin D-resistant rickets / Mutações no gene do receptor de vitamin D em quatro pacientes com raquitismo hereditário resistente a 1, 25-dihydroxyvitamin D

Mutations in the vitamin D receptor (VDR) are associated to the hereditary 1,25-dihydroxivitamin D-resistant rickets. The objectives of this work are: search for mutations in the VDR and analyze their functional consequences in four Brazilian children presented with rickets and alopecia. The coding region of the VDR was amplified by PCR e direct sequenced. We identified three mutations: two patients had the same mutation in exon 7 at aminoacid position 259 (p.Q259E); one patient had a mutation in exon 8 at codon 319 (p.G319V) and another one had a mutation in exon 3 leading to a truncated protein at position 73 (p.R73X). Functional studies of the mutant receptors of fibroblast primary culture, from patients' skin biopsy treated with increasing doses of 1,25(OH)2 vitamin D showed that VDR mutants were unable to be properly activated and presented a reduction in 24-hydroxylase expression level.

Mutações no receptor de vitamina D (VDR) são associadas a raquitismo hereditário resistente a 1,25-dihidroxivitamina D. Os objetivos deste trabalho foram procurar mutações no VDR e analisar suas conseqüências funcionais em quatro pacientes com raquitismo e alopécia. A região codificadora do VDR foi amplificada por PCR e seqüenciada diretamente. Identificamos três mutações: dois pacientes apresentavam a mesma mutação no éxon 7 na posição protéica 259 (p.Q259E); um paciente apresentava uma mutação no éxon 8 no códon 319 (p.G319V) e o outro apresentava uma mutação no exon 3 resultando em uma proteína truncada na posição 73 (p.R73X). O estudo funcional dos receptores mutados nos extratos de culturas de fibroblasto primárias obtidas de biópsia de pele dos pacientes, tratados com doses crescentes de 1,25(OH)2 vitamina D demonstraram que os receptores mutantes não apresentam ativação adequada apresentando expressão reduzida de 24-hidroxilase.

Altered bone metabolic markers in type 2 diabetes mellitus: impact of glycemic control

To study the influence of glycemic control on bone minerals and biochemical markers of bone metabolism in patients with type 2 diabetes mellitus. A Case-control study was conducted at Uhod Hospital, KSA from October 2003 to August 2004 to study 60 premenopausal, multipareous female patients with type 2 diabetes mellitus for >5 years, under oral anti-diabetics, with no diabetic complications. They were divided according to their glycemic control into: controlled group [n=22] and poorly controlled group [n=38] and were compared to 30 age matched healthy women. Osteocalcin [OC], urinary deoxypyridinoline [Dpd], Parathyroid hormone [PTH] were measured by chemiluminescent enzyme immunometric assay and 25 hydroxy vitamin D [25 OH-D] was measured by high performance liquid chromatography. In both diabetic groups, there were higher ALP [177 +/- 39.88 and 287 +/- 41.4 mg/dl] and PTH [49 +/- 9.87 and 56.25 +/- 12.3 Pg/ml] than in controls [144 +/- 22.54 mg/dl, 26.9 +/- 5.60 Pg/ml respectively], but lower serum calcium [8.87 +/- 0.3 and 8.79 +/- 0.7 mg/dl], and 25 OH-D [50.9 +/- 12.6, 45.4 +/- 18.9 micro g/l] and osteocalcin [4.09 +/- 1.48 and 1.89 +/- 0.24 ng/ml] than controls [9.96 +/- 1.9l, 57.9 +/- 13.6 micro g/l, 6.5 +/- 1.5 ng/ml respectively], Urinary calcium and urinary Dpd were higher [270.66 +/- 41.7 and 300.56 +/- 55.67 mg/d and 10.8 +/- 4.6, 12.06 +/- 5.12 nM/mM creatinine] than in controls [244.23 +/- 51.5 mg/d, 6.2 +/- 0.8 nM/mM creatinine]. Glycemic indices [FBG, HbA1C] showed significant positive correlation with ALP [r=0.290 and 0.294], urinary calcium [r=0.340 and 0.260] and Dpd [r=0.468 and 0.228]. Our data give evidence of altered bone metabolic markers in both controlled and uncontrolled female patients with type 2 diabetes mellitus with more significant alterations in the uncontrolled group. This could reflect the strong impact of glycemic control on diabetic bone turnover

Molecular characterization of 21 hydroxylase deficiency congenital adrenal hyperplasia in Egyptian children: a pilot study

Defective steroid synthesis due to derangements of 21-hydroxylase gene [CYP21] constitutes the most frequent cause of congenital adrenal hyperplasia [CAH] and is one of the most frequent inborn errors of metabolism world wide. The molecular basis of CYP21 mutations is complicated. During meiosis gene conversion occurs and transfers deleterious point mutations from the inactive [CYP21P] to the corresponding sequence of the CYP21 gene causing either complete or partial inactivation of 21-hydroxylase activity. Allele specific polymerase chain reaction [ASPCR] was used for the detection of the 4 most common mutations in CYP21 gene: Intron 2 splice mutations [IVS2-13], 8bp deletion in exon 3 [del-8bp], I172N mutation in exon 4 and V281L mutation in exon 7, in 11 salt wasting SW-CAH Egyptian infants. In the examined 22 alleles, 2 alleles were carrying del-8bp, 3 were carrying IVS2-13 mutation, 4 with I172N, 4 with V281L. In the present study the percentage of undetectable mutations was 50%. The wide range of genetic mutations reported for CYP21 gene reconfirm the marked heterogeneity of the disorder among Egyptian and calls for more extensive molecular work

The effects of serum calcium and 1 alpha hydroxylase on the expression of 24-hydroxylase gene / 中国应用生理学杂志

<p><b>AIM</b>To investigate the effects of 1 alpha hydroxylase and serum calcium on the expression of 24-hydroxylase gene in mice kidney.</p><p><b>METHODS</b>Mice with targeted deletion of the 25-hydroxyvitamin D 1 alpha hydroxylase gene(1alpha (OH)ase-/-), and the vitamin D receptor gene (VDR-/-) were used. The study of each mutant had two groups which were (1) mutant with high calcium diet, which maintained fertility but left mice hypocalcaemia; (2) mutant with high lactose diet, which normalized calcium in two mutant. Mice in same litter were as control. There were six groups in total and each group had five mice. All mice were killed at 10-week-old. Serum calcium was determined by an autoanalyzer. RNA was isolated from mouse kidney and the express of 1 alpha hydroxylase gene and 24-Hydroxylase gene were studied by RT-PCR.</p><p><b>RESULTS</b>On the high calcium intake, all mutant animals were hypocalcaemia (1alpha (OH)ase-/- (78 +/- 10.4) mg/L, P < 0.05; VDR-/- (68 +/- 9.8) mg/L, P < 0.05. WT (111 +/- 16.5 mg/L), but when the high lactose diet was administered, serum calcium levels in two mutant mice rose to wild-type levels. The 1 alpha hydroxylase gene was expressed at very higher levels in the vitamin D receptor mutant mice than in wild-type mice when animals received a high calcium intake; This was reduced by eliminating hypocalcaemia with the high lactose diet. Expression of the 24(OH)ase gene was extremely down-regulated in two mutant mice on the high calcium diet but was restored to wild-type levels on the high lactose diet.</p><p><b>CONCLUSION</b>The express of 24-hydroxylase gene was directly regulated by serum calcium rather than 1 alpha-hydroxylase. These studies indicate that both the serum calcium and 1 alpha-hydroxylase exert effects on the expression of 24-hydroxylase gene, but 1 alpha-hydroxylase take the effects by elevated the concentration of serum calcium. There are no direct interaction between 1 alpha-hydroxylase gene and 24-hydroxylase gene.</p>

Cerebrotendinous xanthomatosis: report of two Brazilian brothers

Xantomatose cerebrotendínea é doença autossômica recessiva tratável causada pelo acúmulo de lipídeos por deficiência da enzima 27-esterol hidroxilase na produção de ácido cólico e deoxicólico. Descrevemos dois irmãos brasileiros com dificuldade cognitiva e diarréia crônica. Um deles apresentava catarata bilateral. Os achados neurológicos foram dificuldade progressiva para deambular, ataxia de membros e sinais piramidais. Ambos tinham xantomas de tendão aquileu bilateralmente. O exame de ressonância magnética revelou áreas de sinal hiperintenso em ambos os hemisférios cerebelares. Descrevemos os casos com diagnóstico genético comparando-os com a literatura. O estudo do gene CYP27A1 demonstrou a mutação C1183T no exon 6.

Intracellular calcium ion release stimulated by 1,25(OH)2D3 and influenced with mechanical pressure in mandibular condylar chondrocytes of rabbit / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the intracellular calcium ion release and the system of calcium channel by 1,25 (OH)2D3 stimulus, and the effect of mechanical pressure on it in rabbit mandibular condylar chondrocytes (MCC) in vitro.</p><p><b>METHODS</b>In vitro cultured MCC from two-week-old New Zealand rabbits were incubated under 20 g/L heparin, 1 g/L procaine, continuous pressure of 90 kPa for 60 min and 360 min in a hydraulic pressure controlled cellular strain unit. With the Fluo-3/AM probe loaded, 1,25(OH)2D3 was added to the medium and then the intracellular calcium level was detected by a laser confocal scanning microscope.</p><p><b>RESULTS</b>Intracellular calcium concentration increased in MCC treated with 1,25(OH)2D3, 1,25(OH)2D3 and procaine, while it didn't change in heparin treated group. Calcium in group under continuous pressure of 90 kPa for 60 min was also increased, even higher than the group stimulated only with 1,25(OH)2D3. Intracellular calcium in group treated with continuous pressure of 90 kPa for 360 min showed no significant difference compared to the control and even decreased at the end of the recording period.</p><p><b>CONCLUSION</b>1,25(OH)2D3 could stimulate the intracellular calcium release channel of inositol triphosphate (IP3) receptor open in MCC in vitro and increases the level of intracellular calcium concentration. Pretreatment of definite mechanical pressure could modulate the sensitivity of IP3 channel to 1,25(OH)2D3 stimulus.</p>

Congenital adrenal hyperplasia: biochemical and molecular perspectives.

Congenital adrenal hyperplasia is a disorder occurring in both sexes and is the commonest cause of ambiguous genitalia. It is a group of autosomal recessive disorders in which, on the basis of an enzyme defect the bulk of steroid hormone production by adrenal cortex shifts from corticosteroids to androgens. Autosomal recessive mutations in the CYP21, CYP17, CYP11B1 and 3betaHSD genes that encode steroidogenic enzymes, in addition to mutations in the gene encoding the intracellular cholesterol transport protein steroidogenic acute regulatory protein StAR can cause CAH. Each of the defects causes different biochemical consequences and clinical features. Deficiencies in 21 hydroxylase (21-OH) and 11beta-Hydroxylase (11beta-OH) are the two most frequent causes of CAH. All the biochemical defects impair cortisol secretion, resulting into compensatory hypersecretion of ACTH and consequent hyperplasia of the adrenal cortex. Research in recent years has clarified clinical, biochemical and genetic problems in diagnosis and treatment of the disorders. Expanding knowledge of the gene mutations associated with each of these disorders is providing valuable diagnostic tools in addition to the biochemical profile and phenotype. Genotyping is useful in selecting instances to provide genetic counseling and to clarify ambiguous cases.

Effect of deltamethrin on benzyloxyresorufin O-dealkylatase activity and CYP2B1/2B2 protein level in rat brain / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To detect the effect of deltamethrin (DM) on benzyloxyresorufin O-dealkylatase (BROD) activity and the expression of CYP2B1/2B2 in rat brain.</p><p><b>METHODS</b>BROD activity was determined by fluorophotometry under the treatment of DM in vivo and vitro. Western-blot analysis was used to detect the expression of CYP2B1/2B2.</p><p><b>RESULTS</b>In vivo, DM could markedly inhibit BROD activity in rat brain microsome after rats had been treated with DM (12.5 mg.kg-1.d-1, i.p.) for 5 days. The inhibitory rate in whole brain, cerebral cortex and cerebellum were 26.7%, 23.8% and 33.3% respectively(P < 0.05). However, in vitro, under the concentration of 2 x 10(-8)-2 x 10(-4) mol/L of DM, there was no obvious change of BROD activity in rat brain. Moreover, Western-blot analysis indicated that DM could significantly reduce the expression of CYP2B1/2B2 in vivo, the inhibitory rate of protein synthesis was 42.6%.</p><p><b>CONCLUSION</b>DM could inhibit BROD activity in rat brain and this effect may be related to the reduction of CYP2B1/2B2 protein synthesis.</p>

Screening for late onset congenital adrenal hyperplasia due to 21-hydroxylase deficiency.

Basal and post-ACTH levels of 17 alpha hydroxy-progesterone (17 OHP) were determined in 53 subjects with hirsutism. Late onset congenital adrenal hyperplasia (LOCAH) was detected in five (10.6%) on the basis of elevated basal and/or ACTH stimulated levels of 17 OHP. Of the five patients, two were considered to have a heterogygous state on account of a small rise in stimulated 17 alpha OHP. Screening tests for LOCAH are essential as the clinical diagnosis is not otherwise possible for this treatable and often familial disorder.