Congenital adrenal hyperplasia due to 11-beta-hydroxylase deficiency: description of a new mutation, R384X / Hiperplasia suprarrenal congénita por déficit de 11-beta-hidroxilasa: descripción de una nueva mutación, R384X

Abstract Case Description: It is presented the phenotype of a new compound heterozygous mutation of the genes R384X and Q356X encoding the enzyme of 11-beta-hydroxylase Clinical Findings: Severe virilization, peripheral hypertension, and early puberty. Treatment and Outcome: Managed with hormone replacement therapy (corticosteroid) and antihypertensive therapy (beta-blocker), resulting in the control of physical changes and levels of arterial tension. Clinical Relevance: According to the phenotypic characteristics of the patient, it is inferred that the R384X mutation carries an additional burden on the Q356X mutation, with the latter previously described as a cause of 11-beta-hydroxylase deficiency. The description of a new genotype, as in this case, expands the understanding of the hereditary burden and deciphers the various factors that lead to this pathology as well as the other forms of congenital adrenal hyperplasia (CAH), presenting with a broad spectrum of clinical presentations. This study highlights the importance of a complete description of the patient's CAH genetic profile as well as their parents' genetic profile.

Resumen Descripción del Caso: Se presenta el fenotipo de una nueva mutación heterocigota compuesta en los genes Q356X y R384X que codifican la enzima 11-beta-hidroxilada Hallazgos Clínicos: Virilización severa, pubertad precoz periférica e hipertensión. Tratamiento y Resultados: Manejo con terapia de reemplazo hormonal con corticoide y antihipertensivo con beta-bloqueador con lo que se logró controlar los cambios físicos y los niveles de tensión arterial. Relevancia Clínica: Según las características fenotípicas del paciente se infiere que la mutación R384X acarrea una carga adicional a la mutación Q356X, esta última descrita como causa de deficiencia de 11-beta-hidroxilasa. La descripción de nuevos genotipos, como en este caso, permite ampliar la comprensión de la carga hereditaria y descifrar los diversos factores que llevan a que esta patología, así como las demás formas de hiperplasia suprarrenal congénita (HSC), se presenten con un amplio espectro de cuadros clínicos. Esto permite resaltar la importancia de una descripción completa del perfil genético del paciente con HSC y de sus padres.

Histone deacetylase inhibitor, CG200745, attenuates cardiac hypertrophy and fibrosis in DOCA-induced hypertensive rats

CG200745 is a novel inhibitor of histone deacetylases (HDACs), initially developed for treatment of various hematological and solid cancers. Because it is water-soluble, it can be administered orally. We hypothesized that the HDAC inhibitor, CG200745, attenuates cardiac hypertrophy and fibrosis in deoxycorticosterone acetate (DOCA)-induced hypertensive rats. For establishment of hypertension, 40 mg/kg of DOCA was subcutaneously injected four times weekly into Sprague-Dawley rats. All the rats used in this study including those in the sham group had been unilaterally nephrectomized and allowed free access to drinking water containing 1% NaCl. Systolic blood pressure was measured by the tail-cuff method. Blood chemistry including sodium, potassium, glucose, triglyceride, and cholesterol levels was analyzed. Sections of the heart were visualized after trichrome and hematoxylin and eosin stain. The expression of hypertrophic genes such as atrial natriuretic peptide A (Nppa) and atrial natriuretic peptide B (Nppb) in addition to fibrotic genes such as Collagen-1, Collagen-3, connective tissue growth factor (Ctgf), and Fibronectin were measured by quantitative real-time PCR (qRT-PCR). Injection of DOCA increased systolic blood pressure, heart weight, and cardiac fibrosis, which was attenuated by CG200745. Neither DOCA nor CG200745 affected body weight, vascular contraction and relaxation responses, and blood chemistry. Injection of DOCA increased expression of both hypertrophic and fibrotic genes, which was abrogated by CG200745. These results indicate that CG200745 attenuates cardiac hypertrophy and fibrosis in DOCA-induced hypertensive rats.

Estudo da superposição de mecanismos fisiopatológicos como modelo de hipertensão arterial com repercussões cardíacas graves / Study of overlap of pathophysiologicalmechanisms as a model of hypertensionwith severe cardiac effects

Fundamento: Acredita-se que o rato espontaneamente hipertenso (SHR) mimetize a hipertensão arterial (HA) essencial em humanos. Lesões em órgãos-alvo nesses animais não são devidas unicamente ao aumento da pressão arterial. Outros mecanismos fisiopatológicos superajuntados talvez representem melhor o complexo dano cardiovascular observado também em humanos. Objetivo: avaliar, comparativamente, as alterações cardíacas em ratos SHR nos quais mecanismos outros de HA (renovascular, hipervolemia, disfunção endotelial) sejam superpostos. Materiais e Métodos: cinco grupos foram estudados: Controle (C,n=11); SHR (n=11); SHR + L-NAME (SHR + L-NAME, n=11); SHR com estenose cirúrgica de artéria renal (SHR + 2R-1C, n=11); SHR+ deoxicorticosterona e cloreto de sódio 0,9% (SHR+DOCA-SALT, n = 11). Foram avaliados pressão arterial caudal (PAC), hipertrofia do ventrículo esquerdo (VE) e alterações histológicas miocárdicas. Resultados: Após oito semanas, os grupos SHR + L-NAME, SHR + 2R-1C e SHR + DOCA-SALT mantiveram PAC semelhante e mais elevada que os animais SHR (159,9 ± 8,3; 162,7± 16,7 e 166,3 ± 6,7 versus 138±7,8, respectivamente), bem como a espessura relativa da parede do VE (SHR + L-NAME = 0,64 ± 0,06; SHR+DOCA-SALT=0,63 ± 0,07 versus SHR=0,57±0,03) (p < 0,05). Amassa relativa do VE dogrupo SHR+ L-NAME (4,2±1,15) foi maior que nos demais grupos (SHR=2,8±0,5; SHR + 2R-1C=3,2±0,5; SHR + DOCA-SALT = 3,1 ± 0,2) (p <0,05). O desarranjo de fibras, fibrose intersticial, espessura médio-intimal aumentada foram mais frequentes nos ratos SHR + L-NAME. Conclusão: O modelo SHR + L-NAME mostrou repercussões cardíacas mais evidentes que os demais modelos de HA, fato não explicado apenas pelos níveis de PA elevados. Tal modelo pode ser utilizado em estudos futuros como representativo de maior comprometimento cardíaco na HA grave ou em estados avançados da doença.

Background: Spontaneously hypertensive rats (SHR) are believed to mimic arterial hypertension (HA) essential in humans. The injuries on targeted organs on these animals are not due only to increased blood pressure. Other pathophysiological mechanisms may represent better the superimposed complex cardiovascular damage observed on humans as well. Objective: comparatively evaluate the cardiac abnormalities in SHR in which other mechanisms of hypertension (renovascular, fluid overload and endothelial dysfunction) are superimposed. Materials and Methods: 5 groups were studied: Control (C, n=11); SHR (n = 11); SHR + L-NAME (SHR + L-NAME, n= 11), SHR with surgical stenosis of renal artery (SHR+ 2K-1C, n = 11), SHR + deoxycorticosterone and sodium chloride 0.9% (SHR + DOCA-SALT, n = 11). The blood pressure flow (PAC), left ventricular hypertrophy (VE) (echocardiography) and myocardial histological changes were evaluated. Results:After 8 weeks, the SHR + L-NAME, SHR + 2K-1C and SHR + DOCA-SALT groups maintained similar and higher PAC than SHR (159.9 ± 8.3, 162.7 ± 16.7 and 166.3 ± 6.7 versus 138± 7.8, respectively) as well as the relative thickness of the VE wall (SHR + L-NAME = 0.64 ± 0.06, SHR + DOCA-SALT= 0, 63 ± 0.07 versus SHR = 0.57 ±0.03;) (p < 0.05). The relative LV mass (MRVE, mg/g) of the group SHR + L-NAME (4.2 ± 1.15) was higher than in other groups (SHR = 2.8 ± 0.5, SHR + 2K-1C = 3.2±0.5, SHR + DOCA-SALT = 3.1 ± 0.2) (p < 0.05). Fiber disarray, interstitial fibrosis and the increased of intima-media thickness were more frequent in SHR + L-NAME. Conclusion: SHR + L-NAME showed cardiac effects more evident than the other models of hypertension, which was not explained only by high levels of PA. This model can be used in future studies as representative of greater cardiac involvement in hypertension or severe stages of the disease.

Effect of inhibiting brain reactive oxygen species on sympathetic nerve activity in DOCA-salt hypertensive rats / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate whether brain reactive oxygen species mediate sympathoexcitation and arterial pressure elevation in DOCA-salt hypertensive rats.</p><p><b>METHODS</b>DOCA-salt hypertensive model was established in male SD rats by subcutaneous injection of DOCA after uninephrectomy and drinking 1% NaCl solution for 4 weeks. The baseline mean arterial pressure (MAP), heart rate (HR) and renal sympathetic nerve activity (RSNA) were recorded in the rats under mild anesthesia, and MAP changes following intravenous hexamethonium injection were observed. The responses of MAP, HR and RSNA to intracerebroventricular administration of tempol (20 µmol/L in 10 µl) were evaluated; plasma NE level was measured with ELISA, and ROS level and NAD(P)H oxidase activity in the hypothalamus were detected using chemiluminescence assay.</p><p><b>RESULTS</b>MAP and plasma NE levels were significantly increased in DOCA-salt rats as compared with those in the control group (P<0.01). In DOCA-salt hypertensive rats, intravenous hexamethonium injection induced a blood pressure reduction 240% of that in control rats, and significantly increased the levels of superoxide anion and NAD(P)H oxidase activity in the hypothalamus. Intracerebroventricular microinjection of tempol also resulted in more significant changes of MAP, HR and RSNA in DOCA-salt rats than in the control group (P<0.01).</p><p><b>CONCLUSION</b>Sympathoexcitation due to increased NAD(P)H oxidase-derived ROS levels in the hypothalamus may mediate arterial pressure elevation in DOCA-salt hypertensive rats.</p>

Effects of Tamoxifen in Deoxycorticosterone Acetate-salt Hypertensive Nephropahty

BACKGROUND: The present study was designed to evaluate the possible renoprotective effects of tamoxifen in deoxycorticosterone acetate (DOCA)-salt hypertensive (DSH) rats and its role in inflammation and fibrosis in the kidney. METHODS: Male Sprague-Dawley rats, weighing 180 to 200 g, were used. All rats underwent unilateral nephrectomy. One week later, one group of rats (n = 8) was implanted with DOCA strips (200 mg/kg) and another group of rats (n = 8) was implanted with DOCA strips with co-treated with tamoxifen (10 mg/kg) through gavage feeding. Rats that did not implanted DOCA strips served as controls (n = 6). Two weeks later, the systolic blood pressure (SBP) was measured by tail-cuff method. The protein expression of transforming growth factor-beta (TGF-beta), Smad, alpha-smooth muscle actin (alpha-SMA), E-cadherin, ED-1, cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS) was determined in the kidney by immunoblotting. The mRNA expression of tumor necrosis factor-alpha (TNF-alpha), monocyte chemotactic protein-1 (MCP-1), and vascular cell adhesion molecule-1 (VCAM-1) was determined by real-time polymerase chain reaction. RESULTS: In DSH rats, SBP was increased, which was not affected by tamoxifen treatment. Serum creatinine level was comparable in DSH rats compared with controls, which was not affected by tamoxifen treatment. In DSH rats, the protein expression of TGF-beta, Smad 2/3, Smad 4, alpha-SMA, ED-1, COX-2, iNOS was increased compared with controls, and these changes were attenuated by tamoxifen treatment except that of TGF-beta. The mRNA expression of TNF-alpha, MCP-1, and VCAM-1 was increased, and expression of MCP-1 and VCAM-1 was counteracted by tamoxifen treatment. CONCLUSIONS: Tamoxifen is effective in preventing the progression of nephropathy in DSH rats, the mechanism of which is associated with anti-inflammation and anti-fibrotic effects.

Effects of Tempol on Blood Pressure and Tissue Oxidative Stress in DOCA-alt and L-AME-nduced Hypertension / 대한신장학회잡지

BACKGROUND: Effects of oxidative stress on the development of deoxycorticosterone acetate (DOCA)-salt or N(G)-nitro-L-arginine (L-NAME) hypertension were examined. METHODS: Male Sprague-awley rats were treated with DOCA (200 mg/kg, subcutaneous)-salt or L-NAME (40 mg/L in daily drinking water) for 4 weeks. To reduce the oxidative stress, 4-hydroxyl-2,2,6,6-tetramethylpiperidine-1-oxyl (Tempol, 3 mM/L) was cotreated in drinking water. The expression of endothelial nitric oxide synthase (eNOS) and nitrotyrosine proteins was determined in the renal cortex and thoracic aorta. RESULTS: Tempol prevented the development of DOCA-salt hypertension, whereas it was without effect on L-NAME hypertension. In DOCA-salt hypertension, the eNOS expression in the renal cortex was increased, the degree of which was attenuated by Tempol. The renal expression of nitrotyrosine was decreased, which was further decreased by Tempol. In the aorta, the expression of both eNOS and nitrotyrosine was decreased, which was not further affected by Tempol. In L-NAME hypertension, the renal expression of eNOS was significantly increased, which was blocked by Tempol. The expression of eNOS in the aorta was slightly decreased, and was not further affected by Tempol. The renal expression of nitrotyrosine was not significantly altered. However, its expression was significantly decreased in the aorta, and was further reduced by Tempol. CONCLUSION: The blockade of oxidative stress may attenuate the development of hypertension and provide tissue protection in DOCA-salt hypertension. The blockade of oxidative stress may also contribute to a tissue protection in L-NAME hypertension.

Altered Calcium Current of the Vascular Smooth Muscle in Renal Hypertension

The present study was aimed at investigating whether the calcium current in the vascular smooth muscle (VSM) cells is altered in renal hypertension. Two-kidney, one clip (2K1C) and deoxycorticosterone acetate (DOCA)-salt hypertension were made in Sprague-Dawley rats. Rats without clipping the renal artery or implanting DOCA were used as control for 2K1C and DOCA-salt hypertension, respectively. Four weeks after clipping, systolic blood pressure was significantly higher in 2K1C rats than in control (192+/-24 and 119+/-4 mmHg, respectively, n=16 each). DOCA-salt rats also showed a higher blood pressure (180+/-15 mmHg, n=18) compared with control (121+/-6 mmHg, n=14). VSM cells were enzymatically and mechanically isolated from basilar arteries. Single relaxed VSM cells measured 5 ~ 10 mum in width and 70 ~ 150 mum in length were obtained. VSM cells could not be differentiated in size and shape between hypertensive and normotensive rats under light microscopy. High-threshold (L-type) calciumcurrents were recorded using whole-cell patch clamp technique. The amplitude of the current recorded from VSM cells was larger in 2K1C hypertension than in control. Neither the voltage-dependence of the calcium current nor the cell capacitance was significantly affected by 2K1C hypertension. By contrast, the amplitude of the calcium current was not altered in DOCA-salt hypertension. These results suggest that high-threshold calcium current of the VSM cells is altered in 2K1C hypertension, and that calcium channel may not be involved in calcium recruitment of VSM in DOCA-salt hypertension.

Role of tyrosine kinases in vascular contraction in deoxycorticosterone acetate-salt hypertensive rats

It has been known that activation of tyrosine kinases is involved in signal transduction. Role of the tyrosine kinase in vascular smooth muscle contraction was examined in deoxycorticosterone acetate (DOCA)-salt hypertensive rats. Male Sprague-Dawley rats underwent uninephrectomy, one week after which they were subcutaneously implanted with DOCA (200 mg/kg) and supplied with 1% NaCl and 0.2% KCl drinking water for 4-6 weeks. Control rats were treated the same except for that no DOCA was implanted. Helical strips of carotid arteries were mounted in organ baths for measurement of isometric force development. Genistein was used as a tyrosine kinase inhibitor. Concentration-response curves to 5-hydroxytryptamine (5-HT) shifted to the right by genistein in both DOCA-salt hypertensive and control rats. Although the sensitivity to genistein was similar between the two groups, the maximum force generation by 5-HT was less inhibited by genistein in arteries from DOCA-salt hypertensive rats than in those from controls. Genistein-induced relaxations were attenuated in arteries from DOCA-salt rats. Genistein affected the contraction to phorbol 12, 13-dibutyrate (PDBu) neither in DOCA-salt nor in control arteries. These observations suggest that tyrosine kinase is involved in 5-HT-induced vascular contraction, of which role is reduced in DOCA-salt hypertension.

Congenital Adrenal Hyperplasia with 21-hydroxylase Deficiencies in Twins

Congenital adrenal hyperplasia is inherited disorder of adrenal steroidogenesis. 21-hydroxylase deficiency is the most commone enzymatic defect and is divided into classic and late-onset or nonclassic forms. Both classic non-classic 21-hydrozylase deficiencies are inherited in a recessive manner as allelic variants. But it is rare that happened in twin infants. Chief complaints of affected twins in our case were ambiguous genitalia, hyperpigmentation and dehydrations. They were revealed into hyponatremia, hyperkalemia and increased amount of serum progesterone, 17-hydroxyprogesterone and urinary 17-ketosteroid excretion and were administered with DOCA, 9alpha-fluorohydrocortisone, hydrocortisone to control the electrolyte imbalance. And now, both of them are going to normal ratio of weight gain and body growth.

The Effects of Castration and Sex Hormone Administration on Lactic and Malic Dehydrogenase Activities in the Testis and the Prostate Tissues of Male Rabbits / 대한비뇨기과학회지

As a process to study the mechanism of steroid hormones at the molecular level,the activities of lactic dehydrogenase (L.D.) and malic dehydrogenase (M.D.),NAD-linked transhydrogenases, were measured in the testis and the prostate. Ahundred male rabbits were divided into ten group as follows: Group 1: Control Group 2: Estrogen (6,000 units) injected Group 3: Androgen (1,200 unite)injected Group 4: Progesterone (1,200 units) injected Group 5: Hydrocortisone(30 mg) injected Group 6: DOCA (6 mg) injected Group 7: Castration control Group8: Castration and estrogen (6, OOO units) injected Group 9: Castration and androgen (1,200 units) injected Group 10: Castration and progesterone (1,200units) injected Changes in the activities of lactic dehydrogenase and malic dehydrogenase in theorganic tissues by exogenous steroid hormones were carefullyobserved. The lactic dehydrogenase activities were measured by the method of Wroblewski and La Due, and malic dehydrogenase activities by the Bodansky's modification of Porter's method. The results are as follows: 1) The control valueof L.D. activities in the testicular tissue of normal rabbits proved to be 89,400units per ram. The L.D. activities showed an increase up to 110.4 per cent in theestrogen injected group, 179.3 per cent in the androgen injected group and 147.0 per cent in the progesterone injected group, while the administration of hydrocortisone and DOCA decreased the value down to 85.2 per cent and 81.5 per cent, respectively. 2) The control value of M.D. activities in the testicular tissue of the normal rabbits was 23,600 units per gram. The M.D. activities showed an increase upto 111.4 per cent in the estrogen injected group. 191.1 per cent in the androgen injected group, and 156.8 per cent in the progesterone injected group, while the administration of hydrocortisone and DOCA decreased the value down to 85.2 per cent and 81.5 per cent, respectively. 3) In the prostate tissues of non-castrated rabbits, the L.D. activities were estimated normally to be 48,100 units per gram. The administration of sex hormone resulted in raising the activities upto 101.8 per cent in the estrogen injected group, 196.9 per cent in the androgen injected group and 153.9 per cent in the progesterone injected group. But the administration of hydrocortisone and DOCA decreased the value down to 92.5 per cent and 97.1 per cent, respectively. 4) In the prostate tissue of non-castrated rabbits, the control value of M.D. activities proved to be 14,600 unite per gram. The M.D. activities showed an increase upto 117.8 per cent in the estrogen injected group, 206.8 per cent in the androgen injected group and, 176.7 per cent in the progesterone injected group, while the administration of hydrocortisone and DOCA decreased the value down to 81.9 per cent and 95.2 per cent, respectively. 5) The prostatic L.D. activities were decreased to half the normal two weeks after castration. The administration of sex hormones (i.e., estrogen, androgen and progesterone) acted inclusively upon elevating the level f activities. Androgen, in general, was the most effective to restore the activity to the level of pre-castrated state. 6) The prostatic M.D. activities were also decreased to half the normal two weeks after castration. The administration of sex hormones acted inclusively upon elevating the level ofthe activities. Androgen had a remarkable effect in elevating the M.D. activities, which showed twice the precastration level. In this study, it is concluded that L.D. and M.D. activities are present in the testis and the prostate. They are induced and activated by the administration of sex hormones, of which androgen has the most remarkable effect, and estrogen and progesteronehave less effect, while hydrocortisone and DOCA are ineffective in some cases orinhibitory in others.