Numerous epidemiological studies show a strong association between low birth weight and later life hypertension and metabolic disease. Excessive in utero exposure to glucocorticoids (‘stress hormones’) has been hypothesized to be important in such developmental ‘programming’, acting via crucial physiological, gene expression or structural changes in the developing fetus. Normally, the fetus is protected from the high levels of maternal glucocorticoids by an enzymic placental barrier, 11 betahydroxysteroid dehydrogenase type 2 (11β-HSD2). In the placenta, 11β-HSD2 efficiently converts active maternal glucocorticoids (cortisol in humans; corticosterone in rodents) to physiologically inactive 11-keto forms. In previous studies in rats, maternal administration of dexamethasone, a synthetic glucocorticoid which is minimally metabolized by 11β-HSD2, or carbenoxolone, a potent inhibitor of 11 β-hydroxysteroid dehydrogenase, increased glucocorticoid load to the fetus. This resulted in lower offspring birthweight and later life hypertension and hyperglycemia — important components of the metabolic syndrome. These programming effects were seen when dexamethasone was administered selectively during the third week of gestation. We have used this well-validated model of programming to dissect the molecular mechanisms that mediate the programming of hypertension. In accord with previous observations, administration of dexamethasone (100μg/kg/day) to pregnant rats during the last week of pregnancy significantly reduced offspring birthweight by 10%. Moreover, the 9 month-old adult offspring had systolic hypertension (9% rise) accompanied by significant hypokalemia (10% fall K+). The coexistence of hypertension and hypokalemia suggested that prenatal overexposure to dexamethasone might increase mineralocorticoid activity in the kidney. Intriguingly, although offspring of dexamethasone-treated dams had 46% lower plasma renin concentrations (consistent with intravascular fluid volume expansion), 24-hour total urinary aldosterone levels were significantly reduced compared to controls (reduction of 56%). Maternal dexamethasone treatment was associated with a permanent decrease in 11β- HSD2 mRNA and activity in the kidney of the offspring (45% and 36% respectively). 11β-HSD2 plays an important role in regulation of renal sodium reabsorption (and thereby blood pressure) by acting as a pre-receptor barrier to MR access, preventing glucocorticoids from activating MR in the distal nephron. Thus, the decrease in renal 11β-HSD2 activity would allow greater endogenous glucocorticoids to activate MR, likely accounting for the low-renin, low-aldosterone hypokalemic hypertensive phenotype observed in these offspring. Other components of mineralocorticoid or glucocorticoid signaling pathways, including mineralocorticoid receptor (MR), glucocorticoid receptor (GR) and 11-beta hydroxysteroid dehydrogenase type 1 (11β-HSD1) were not altered in the offspring kidney by prenatal glucocorticoid exposure. Dexamethasone-programmed offspring also showed exaggerated mineralocorticoid activity with increased kalliuresis in response to exogenously administered corticosterone, suggesting that the decrease in renal 11β-HSD2 is functionally important. In this respect, our rat model resembles the syndrome of apparent mineralocorticoid excess where reduced 11β-HSD2 allows illicit activation of MR by glucocorticoids, resulting in excessive sodium reabsorption, hypertension and hypokalemia. We also studied the effects of maternal dexamethasone on offspring erythropoietin expression in the kidney. This followed from previous observations that identified the hepatocyte nuclear factor 4 alpha (HNF4α) as a key gene up-regulated in dexamethasone-programmed offspring liver, where it might be involved in mediating hyperglycemia. HNF4α is also expressed in the kidney. The role of HNF4α in the kidney is not fully understood, but has been implicated in regulation of erythropoietin synthesis. As in the liver, prenatal exposure to dexamethasone caused a significant increase (64% increase) in renal HNF4α expression. The increase in renal HNF4α mRNA was observed early (in one week old offspring) and persisted into adulthood. This was associated with significantly elevated levels of erythropoietin in circulation (110% increase). Moreover, animals that were exposed to prenatal dexamethasone had significantly increased red blood cell mass (7% increase), presumably as a result of upregulation of erythropoietin.

L'administration chronique de rHuEPO peut engendrer de graves effets secondaires. Une augmentation de l’hématocrite provoquée par la rHuEPO, en augmentant l'érythrocytose, la viscosité sanguine et les forces de cisaillement à la surface vasculaire, peut être responsable d'hypertension artérielle (HTA) et de thromboses artérielles. La présence d'une fonction endothéliale normale et de monoxyde d'azote (NO) peut contrer les effets délétères thrombogène et hypertenseur de l'EPO. Sur ces bases, nous avons étudié les effets cardiovasculaires d'une administration chronique de rHuEPO dans différentes situations : dans le cadre du dopage, chezdes rats "sportifs" présentant une dysfonction endothéliale NO-dépendante induite par l'administration chronique de L-NAME et dans le cadre d'un traitement chez des rats urémiques développant une dysfonction endothéliale NO-dépendante résultante d'une néphrectomie de 5/6de la masse rénale. Chez nos rats entrainés, dopés et traités au L-NAME, nous avons observé une altération de la performance physique avec une mortalité importante (51%). Une HTA sévères'est développée chez ces rats, avec des valeurs de pression artérielle (> 220 mmHg) bien plusélevées que celles des rats recevant le L-NAME seul, associée à une altération de la vasorelaxation NO-dépendante aortique (< 60%). Les rats insuffisants rénaux (IRC) ont eux aussi montré une augmentation de la pression artérielle et une dysfonction endothéliale en réponse àl'acétylcholine au niveau de l'aorte et en réponse à une élévation du flux au niveau de l'artère mésentérique perfusée. Ces différents paramètres ont été améliorés par l'exercice. Les coupes de rein colorées au rouge Sirius ont montré une fibrose accentuée chez les rats CKD. La fibrose, la créatinémie et l'albuminurie ont été diminuées par l'exercice seul mais ont été aggravées chez les rats du groupe CKD+EPO+Ex. L'activité NADPH oxydase et l'expression des Nox4, p67phox etMAPK erk1/2 ont été augmentées chez les les rats CKD. L'exercice ou la rHuEPO ont prévenuces augmentations. Cependant, l'activité de la NAD(P)H oxydase et l’expression des MAPKerk1/2 sont restées élevées dans le rein des rats CKD+EPO+Ex. Nos données suggèrent que l'exercice seul a un effet protecteur contre les dysfonctions vasculaire et rénale et la fibrose rénale. Ces effets protecteurs sont associés à une inhibition de l'activité de la NADPH oxydase et des voies de signalisation MAPK erk1/2. Par contre, l'exercice combiné avec le traitement rHuEPO, a des effets délétères sur la structure et la fonction rénale des rats CKD. Ces effets nocifs semblent liés à la stimulation de la NADPH oxydase et des voies de signalisation MAPKerk1/2. Malgré les effets protecteurs cardiovasculaire et rénal de l'entraînement physique, ces résultats mettent en évidence que la fonction rénale peut être potentiellement endommagée ainsique la structure du rein en combinant l'exercice avec le traitement rHuEPO dans l'insuffisance rénale. En conclusion, nous pouvons dire que la rHuEPO affecte gravement la fonction cardiovasculaire du rat entraîné présentant une dysfonction endothéliale. Ce risque étant fatal,beaucoup de sportifs, voulant augmenter leur performance, mettent leur vie en danger. Par ailleurs, ayant remarqué les effets délétères au niveau rénal, en associant exercice et traitement rHuEPO dans des conditions expérimentales sur un modèle d'insuffisance rénale, nous suggérons une investigation clinique afin de vérifier la transposition de nos résultats aux patients insuffisants rénaux. / The chronic administration of rHuEPO can engender side effects. An increase of the hematocritinduced by rHuEPO, by increasing the erythrocytosis, the blood viscosity and the shear stress onvascular surface, can be responsible of arterial high blood pressure and arterial thrombosis. Thepresence of a normal endothelial function and nitric oxide (NO) can counter the noxious effectsof rHuEPO. On these bases, we studied the cardiovascular effects of a chronic administration ofrHuEPO in various frames: within doping field, to trained rats with L-NAME-induced NOdependentendothelial dysfunction and within the framework of a treatment, to chronic kidneydisease (CKD) rats developing endothelial dysfunction caused by the "5/6 nephrectomy". In ourdoped rats, we observed an important mortality (51%). A severe arterial high blood pressuredeveloped in these rats (> 220 mmHg) associated with an impairment of the NO-dependentvasorelaxation (< 60 %). CKD rats also showed an increase in blood pressure and an endothelialdysfunction, in response to acetylcholine in the aorta and in response to a rise in flow in perfusedmesenteric artery. These parameters were improved by exercise. Kidney sections stained withSirius red showed marked fibrosis in CKD rats. Fibrosis, creatinine and albumin were decreasedby exercise alone but were increased in rats from the CKD + EPO + Ex group. NAD(P)H oxidaseactivity and the expression of Nox4, p67phox, and MAPK erk1/2 were increased in CKDrats. Exercise or rHuEPO prevented these increases. However, the NAD(P)H oxidase activityand the expression of MAPK erk1/2 remained high in the kidney of rats from the CKD+EPO+Exgroup. Our data suggest that exercise alone has a protective effect against vascular and renaldysfunction and renal fibrosis. These protective effects are linked to the downregulation of theNADPH oxidase activity and MAPK erk1/2 signaling pathways. However, exercise combinedwith rHuEPO treatment has deleterious effects on kidney structure and function in CKDrats. These adverse effects appear to be related to the stimulation of NADPH oxidase and MAPKerk1/2 signaling pathways. Despite the cardiovascular and renal protective effects of physicaltraining, these results highlight the potentially damaging renal function and structure bycombining exercise with rHuEPO therapy in renal failure. In conclusion, we can say that therHuEPO affects seriously cardiovascular function in trained rat with endothelial dysfunction. Thisrisk being fatal, many sportsmen, looking to increase their performance, put their life in danger.Moreover, having noticed the deleterious effects in the kidney by combining exercise andrHuEPO therapy under experimental conditions on a model of renal failure, we suggest a clinicalinvestigation to verify the transposition of our results to patients with renal failure

Introdução: No processo de remodelamento miocárdico ocorre hipertrofia de miócitos e deposição exacerbada de colágeno no interstício, promovendo alteração na geometria e na função do coração. A eritropoetina (EPO) tem sido amplamente estudada nesse cenário, pois exerce efeitos cardioprotetores. Objetivo: Avaliar o papel da EPO na atenuação do remodelamento estrutural, geométrico e funcional do coração, em modelo experimental de infarto do miocárdio. Materiais e Métodos: Estudados 60 ratos Wistar machos divididos em 4 grupos (Controle; Controle+EPO; Infartado; Infartado+EPO). A fração do volume de colágeno intersticial do ventrículo esquerdo (FVCI-VE) e do ventrículo direito (FVCI-VD) foi determinada em cortes histológicos, corados com picrosirus red utilizando-se o programa QWIN Image Processing and Analysis Software (Leica Microsystems Cambridge Ltd.). Essas mesmas lâminas e o software foram utilizados para a medida do tamanho da área de infarto. A análise anatômica e funcional foi realizada por ecocardiograma, avaliando-se a fração de encurtamento do VE (FE) e o diâmetro diastólico do VE (DDVE). Para o estresse oxidativo, dois kits comerciais foram utilizados na determinação da glutationa e do ADMA. A sobrecarga ventricular, apoptose e inflamação foram realizadas por PCR, em tempo real. Na avaliação da angiogênese, utilizamos a técnica de imunohistoquímica. A análise hematológica foi realizada por exames laboratoriais para dosagem de hemoglobina e hematócrito. Resultados: A FVCI-VE (%) foi maior nos grupos infartados em relação aos grupos controles (p < 0,001), e atenuada pela EPO (p < 0,001, IAM vs IAM+EPO) (CT = 0,76 ± 0,21; CT+EPO = 0,63 ± 0,15; IAM+EPO = 1,43 ± 0,92; IAM= 3,47 ± 2,5). A FVCI-VD (%) também foi maior nos infartados em relação aos grupos controles (CT = 0,60 ± 0,2; CT+EPO = 0,83 ± 0,3; IAM+EPO = 1,01 ± 0,55; IAM = 1,60 ± 1,15) (p < 0,001), mas sem diferença estatística quando comparados os grupos IAM vs IAM+EPO. A EPO não influenciou o tamanho do IAM. Os grupos infartados tiveram piora na fração de encurtamento em relação aos controles (CT = 45,65% ± 6,4; CT+EPO = 40,81% ± 4,44; IAM+EPO = 17,32% ± 6,01 e IAM = 20,11% ± 9,49) (p < 0,001), mas sem proteção da EPO. Os grupos infartados também tiveram maior dilatação do VE (p < 0,001) (CT = 0,73 ± 0,06; CT+EPO = 0,74 ± 0,05; IAM+EPO = 0,81 ± 0,09; IAM = 0,87 ± 0,11) sem atenuação da EPO. Os marcadores de estresse oxidativo, ADMA e glutationa, não evidenciaram ação da EPO nessa via. No que se refere à sobrecarga ventricular, o gene BNP apresentou maior expressão nos grupos infartados, em relação aos controles, porém não foi atenuado pela EPO (p = 0,103). Com relação à apoptose, os genes Bcl-2 e p53 mostraram-se mais expressos nos grupos infartados em comparação aos controles (p < 0,05), mas o Bcl-2 não foi ativado e nem o p53 inibido pela EPO. Os genes estudados na avaliação da inflamação foram TNF-alfa, TGF-beta1 e Ccr-5, também não demonstraram efeito anti-inflamatório da EPO. A semiquantificação da angiogênese pela marcação do VEGF também não apresentou diferença significativa entre os grupos (p = 0,95). A análise da hemoglobina e do hematócrito apresentou diferença significativa em relação aos grupos tratados ou não com EPO (p = 0,003 e p = 0,001), respectivamente. Conclusões: EPO atenuou significativamente o acúmulo de colágeno intersticial, mas não protegeu o coração quanto à dilatação, à disfunção e à sobrecarga ventricular neste modelo. Na fase crônica do infarto, avaliada neste estudo, a EPO não modulou as vias da apoptose, estresse oxidativo e inflamação / Introduction: The process of myocardial remodeling include inappropriate collagen deposition in the interstitium developing an overall process of structural and geometric remodeling of the heart. Erythropoietin (EPO) may have a cardioprotective effects including inflammatory and oxidative stress modulation. Objective: The aim of this study was to assess the role of EPO upon structural, geometric and functional remodeling at the heart. Materials and Methods: 60 Wistar rats were divided into 4 groups: Control, Control+EPO, Infarcted, Infarcted+EPO. Interstitial collagen volume fraction in the left (LV-ICVF) and right ventricle (RV-ICVF) was quantified by videomorphometry using a QWIN Image Processing and Analysis Software (Leica Microsystems Cambridge Ltd.). These same slides and software were also used to measure the size of the infarct area. The analyzed echocardiographic parameters were the left ventricle shortening fraction (LVFS) and diastolic diameter (LVDD). For oxidative stress, two commercial kits were used in to quantify ADMA and glutathione. RT-PCR was used to assess ventricular overload, apoptosis and inflammatory cytokines. For angiogenesis we used immunohistochemistry and hematological analysis was performed by laboratory tests for hemoglobin and hematocrit. Non parametric analysis was performed and p <=0.05 was considered significant. Results: LV-ICVF (%) was greater in the infarcted groups compared to controls (p < 0.001), and attenuated by EPO (p = 0.05, MI vs MI+EPO) (CT = 0.76 ± 0.20; CT+EPO = 0.62 ± 0.16; MI+EPO = 1.22 ± 0.86; MI = 3.80 ± 2.6). The RV-ICVF (%) was also greater in the infarcted groups compared to controls (CT = 0.60 ± 0.2; CT+EPO = 0.82 ± 0.28; MI+EPO = 1.02 ± 0.58; IAM = 1.62 ± 1.20) (p = 0.007) but without statistical difference between MI vs MI+EPO. Regarding infarct size we did not observe any difference. The infarcted groups had a worsening shortening fraction compared to controls (CT = 45.65% ± 6.4; CT+EPO = 40.81% ± 4.44; MI+EPO = 17.32% ± 6.01 and MI = 20.11% ± 9.41) (p < 0,001), but without EPO protection. The infarcted groups also showed increased LV dilation (p < 0.001) (CT = 0.73 ± 0.06; CT+EPO = 0.74 ± 0.05; MI+EPO = 0.81 ± 0.08; MI = 0.90 ± 0.11) without EPO attenuation. Oxidative stress markers ADMA and glutathione did not show EPO action in this pathway. The BNP that evaluate ventricular overload, presented increased expression in infarcted groups (p = 0.04), but not attenuated by EPO (p = 0.103). The genes of apoptosis Bcl-2 and p53 were more expressed in infarcted groups when compared to controls (p < 0.05), but Bcl-2 was not activated and p53 inhibited by EPO. For Inflammation just 3 genes exhibit expression with statistical differences between groups (TGF-beta1, TNF-alfa, and CCr-5), but it did not show the EPO anti-inflammatory effect. The semi-quantification of angiogenesis by VEGF expression also did not show statistically significant differences between groups (p = 0.95).The analysis of hemoglobin and hematocrit presented significant differences compared to groups treated or not with EPO (p = 0.003 and p = 0.001), respectively. Conclusions: EPO significantly attenuated the accumulation of interstitial collagen, but it did not reflected in the protection of the heart dilation or dysfunction and oxidative stress, ventricular overload, apoptosis and inflammation of gene expression in this model

Introdução: Este estudo teve como objetivo avaliar o efeito da interleucina 6 (IL-6) e eritropoetina (EPO) na lesão medular aguda experimental em ratos. Materiais e Métodos: A lesão foi induzida pelo equipamento padronizado NYU Impactor, com queda de um peso de 10 g à distância de 12,5 mm de altura. Foram utilizados 50 ratos da linhagem Wistar, divididos em cinco grupos de 10 animais. O grupo EPO, ratos tratados com EPO; grupo EPO/IL-6, animais tratados com EPO e IL-6; grupo IL-6, administração de IL-6; grupo placebo, solução placebo; grupo Sham, procedimento sham (apenas laminectomia, sem lesão medular). Todas as drogas e soro fisiológico foram administrados por via intraperitoneal, durante três semanas. Os animais foram acompanhados por 42 dias. A recuperação motora funcional foi monitorada pela escala de Basso, Beattie e Bresnahan (BBB) nos dias 2, 7, 14, 21, 28, 35 e 42. Exame de potencial evocado foi efetuado no 42o dia, sendo realizada análise histológica qualitativa e quantitativa após eutanásia. Resultados: Os resultados das avaliações da escala BBB mostraram recuperação funcional motora superior no grupo que recebeu EPO. A administração de IL-6 isolada não mostrou benefícios em relação ao grupo que recebeu solução placebo e a associação de IL-6 com EPO mostrou resultados inferiores ao grupo que recebeu apenas EPO. Conclusão: Concluímos que uso EPO após lesão medular contusa em ratos mostrou benefícios na recuperação motora. A associação de EPO e IL- 6 mostrou benefícios, porém com resultados inferiores aos da EPO isolada. O uso isolado de IL-6 não mostrou benefícios após lesão medular contusa experimental em ratos / Introduction: The aim of this study was to evaluate the effect of interleukin-6 (IL-6) and erythropoietin (EPO) in experimental acute spinal cord injury in rats. Methods: The injury was induced by a standardized equipment for spinal cord contusion injury, the NYU Impactor, which produced the lesion by means of a 10g weight drop on the animals\' spinal cord from a 12.5-mm height. Fifty Wistar rats were divided in five groups of ten animals: Group 1 rats treated with EPO; Group 2 animals treated with EPO and IL-6; Group 3, IL-6 administration; Group 4, placebo solution; Group 5, sham procedure (only laminectomy, without spinal cord injury). All drugs and placebo solution were administered intraperitoneally for three weeks. The animals were followed up for 42 days. The functional motor recovery was monitored by the scale of Basso, Beattie and Bresnahan (BBB) on days 2, 7, 14, 21, 28, 35 and 42. Evoked potential tests were performed on the 42nd day. Qualitative and quantitative histological analysis were performed after euthanasia. Results: The group receiving EPO demonstrated superior functional motor results in the BBB scale. IL-6 administration alone did not show benefits over the placebo group solution and the IL-6 combination with EPO showed results lower than those seen in the group that received only EPO. Conclusion: We conclude that using EPO after acute spinal cord injury in rats showed benefits in motor recovery. The association of EPO and IL-6 showed benefits, but with inferior results to the isolated EPO. Isolated use of IL-6 showed no benefit after experimental spinal cord injury in rats

As síndromes talassêmicas (?- e ?-talassemia) são as desordens mais comuns e frequentes associadas com eritropoese ineficaz. O desbalanço na produção das cadeias ?- e ?-globinas resulta no comprometimento da produção de eritrócitos, em anemia e aumento de progenitores eritroides no sangue periférico. Enquanto os pacientes homozigóticos afetados por essas desordens demonstram alterações características dos parâmetros relacionados a eritropoese, a relação entre grau de anemia, eritropoese alterada e disfunção do metabolismo de ferro ainda não foram investigados nos indivíduos com ?+-talassemia heterozigótica ou ?+-talassêmia. Duzentos e vinte seis indivíduos (75 do gênero feminino e 151 do gênero masculino) foram recrutados e divididos em 5 grupos: Controle (n=28), doadores de sangue regulares (DSR, n=23), ?+-talassemia heterozigótica (TAT, n=14), ?+-thalassemia (traço ?-talassêmico, TBT, n=20) e ?0-talassemia, (?-talassemia maior, BTM, n=27). As amostras foram analisadas para parâmetros hematológicos (Micros ABX 60); ferro sérico, capacidade total de ligação ao ferro e saturação de transferrina por método colorimétrico (Pointe Scientific, Inc., Canton, MI, USA), ferritina e proteína C-reativa ultra sensível por imunoensaio (Immulite 1000); receptor solúvel de transferrina, eritropoetina, fator de diferenciação do crescimento 15 (R&D Systems) e hepcidina (Intrinsic LifeSciences, La Jolla, CA) por ELISA. As razões sTfR/log ferritina e (hepcidina/ferritina)/sTfR foram calculadas para avaliar o metabolismo do ferro. sTfR/log ferritina pode distinguir depleção dos estoques de ferro de eritropoese deficiente de ferro, enquanto (hepcidina/ferritina)/sTfR pode avaliar os estímulos contrários (disponibilidade de ferro e atividade eritropoética) que controlam a síntese de hepcidina e a absorção de ferro, na ausência de estímulos inflamatórios. Foi demonstrado que TAT teve significativa redução da hepcidina e aumento do receptor solúvel de transferrina, com parâmetros hematológicos relativamente normais. Em contraste, todos os parâmetros hematológicos de TBT foram significativamente diferentes do Controle, incluindo aumento dos níveis do receptor solúvel de transferrina, ferritina, eritropoetina e fator de diferenciação do crescimento 15. Essas alterações em ambos os grupos sugerem um balanço alterado entre eritropoese e metabolismo de ferro. Os índices sTfR/log ferritina e (hepcidina/ferritina)/sTfR estão, respectivamente, aumentado e reduzido comparados ao Controle, proporcional a severidade de cada grupo talassêmico. Em conclusão, destacamos que, pela primeira vez, foram descritas alterações no metabolismo de ferro em indivíduos com ?+-talassemia heterozigótica. Esses dados demonstram que, no contexto da saúde pública, são necessários identificação e acompanhamento dos portadores de ?+-talassemia. / The thalassemia syndromes (?- and ?-thalassemia) are the most common and frequent disorders associated with ineffective erythropoiesis. Imbalance of ?- or ?-globin chain production results in impaired red blood cell synthesis, anemia and more erythroid progenitors in the blood stream. While patients affected by these disorders show definitive altered parameters related to erythropoiesis, the relationship between the degree of anemia, altered erythropoiesis and dysfunctional iron metabolism have not been investigated in both carriers of ?-thalassemia and ?-thalassemia. 226 subjects (75 females and 151 males) were recruited to this study and divided in 5 groups: Control (n=28), repeat blood donors (DSR, n=23), ?+-thalassemia heterozygous carriers (TAT, n=14), ?+-thalassemia (?-thalassemia trait, TBT, n=20) and ?0-thalassemia, (?-thalassemia major, BTM, n=27). Samples were tested for hematological parameters (Micros ABX 60); serum iron, total iron binding capacity, and transferrin saturation by the colorimetric method (Pointe Scientific, Inc., Canton, MI, USA), ferritin and high sensitive C-reactive protein by immunoassay (Immulite 1000); soluble transferrin receptor, erythropoietin and growth differentiation factor 15 (R&D Systems) and hepcidin (Intrinsic LifeSciences, La Jolla, CA) by ELISA. Were calculated the ratios sTfR/log ferritin and (hepcidin/ferritin)/sTfR to evaluate iron metabolism. sTfR/log ferritin can distinguish storage iron depletion from iron-deficient erythropoiesis, while (hepcidin/ferritin)/sTfR can be utilized to explore and quantify the opposing forces (i.e. iron availability and erythropoietic activity) regulating hepcidin synthesis and iron absorption in absence of inflammatory stimuli. We demonstrate that TAT have a significantly reduced hepcidin and increased soluble transferrin receptor levels but relatively normal hematological findings. In contrast, TBT have all hematological parameters significantly different from controls, including increased soluble transferrin receptor, ferritin, erythropoietin and growth differentiation factor 15 levels. These changings in both groups suggest an altered balance between erythropoiesis and iron metabolism. The indexes sTfR/log ferritin and (hepcidin/ferritin)/sTfR are respectively increased and reduced relative to controls, proportional to the severity of each thalassemia group. In conclusion, we emphasize that, for the first time in the literature, subjects with heterozygous ?+-thalassemia have altered iron metabolism. Our data demonstrate that within the context of public health, identification and monitoring of patients with ?+-thalassemia are needed.

Os efeitos farmacológicos da eritropoetina (EPO) e do tacrolimo (FK 506) têm sido investigados no tratamento da lesão medular, mas são escassos os trabalhos que avaliam a interação entre essas drogas. Neste estudo experimental, 60 ratos Wistar foram submetidos a lesão contusa da medula espinal produzida pelo sistema NYU Impactor. Os animais foram divididos em cinco grupos, sendo: Controle, que recebeu soro fisiológico; EPO, que recebeu eritropoetina; o EPO + FK 506 recebeu EPO associada ao tacrolimo; o FK 506 recebeu tacrolimo. Todas as drogas e o soro fisiológico foram administrados por via intraperitoneal. O grupo Sham foi submetido à lesão medular, mas não recebeu nenhuma droga. Os animais foram avaliados quanto à recuperação da função locomotora em sete diferentes momentos pelo teste de BBB no 2o, 7o, 14o, 21o, 28o, 35o e 42o dias após lesão contusa na medula espinal. No 42o dia, foi realizada avaliação eletrofisiológica dos animais que, logo após, foram sacrificados para análise dos achados histológicos da medula lesionada. Nosso projeto experimental não revelou diferenças na recuperação da função locomotora, nas análises histológica e eletrofisiológica nos animais submetidos ao tratamento farmacológico com eritropoetina e com tacrolimo, após contusão medular torácica / The pharmacological effects of erythropoietin (EPO) and tacrolimus (FK 506) have been investigated in the treatment of spinal cord injuries, but there are few studies that evaluate the interaction between these drugs. In this experimental study, 60 Wistar rats were submitted to contusion spinal cord injury produced by the NYU Impactor system. The animals were divided into five groups: Control, which received saline only; EPO, which received erythropoietin; EPO + FK 506, which received EPO associated with tacrolimus; and the group FK 506, which received tacrolimus. All drugs and saline were administered intraperitoneally. The Sham group underwent spinal cord injury, but did not receive any drug. The animals were evaluated for recovery of locomotor function in seven different times by the BBB test, in the 2nd, 7th, 14th, 21st, 28th, 35th and 42nd days after spinal cord injury. In 42 days, electrophysiological evaluation was performed, and the animals were, shortly after, sacrificed for histological analysis of the injured spinal cord. Our experimental study did not reveal significant differences in the recovery of locomotor function, nor in the histological and electrophysiological analysis in animals treated with erythropoietin and tacrolimus after thoracic spinal cord injury

A sepse envolve mecanismos complexos de respostas imunológicas e inflamatórias, e o papel do NF- B é essencial. A diminuição da NO sintase endotelial (eNOS) durante a sepse contribui com a disfunção endotelial. A eritropoietina (EPO) é uma citocina protetora de diversos tecidos durante o estresse. Investigamos o papel da EPO na injúria renal aguda (IRA) induzida pela sepse usando o modelo de ligadura e punção do ceco (LPC). Ratos Wistar foram divididos em três grupos: controle; LPC e LPC+EPO (EPO, 4.000UI/kg, administrada 24h e 1h antes da cirurgia). Com a finalidade de estudar os efeitos precoces e tardios da EPO sobre a IRA induzida pela sepse realizamos três etapas de experimentos: Primeira etapa: 24 horas após LPC; Segunda etapa: 48 horas após LPC; Terceira etapa: análise de sobrevida. No estudo precoce o grupo LPC+EPO apresentou clearance de inulina significativamente maior que o grupo LPC. Recuperou os níveis de hematócrito na sepse, melhorou a pressão arterial e a acidose metabólica. No estudo tardio o grupo LPC+EPO apresentou clearance de creatinina significativamente maior que o grupo LPC. Nesta fase tardia a EPO recuperou os níveis de eNOS, suprimiu a infiltração de macrófagos no tecido renal e inibiu a ativação do NF- B. A EPO protege a função renal e aumenta a sobrevida neste modelo de sepse. A proteção da EPO na sepse é dependente, em parte, da inibição do NF- B e do aumento da expressão de eNOS / The pathophysiology of sepsis involves complex cytokine and inflammatory mediator networks, a mechanism to which nuclear factor-kappa B (NF- B ) activation is central. Downregulation of endothelial nitric oxide synthase (eNOS) contributes to sepsis-induced endothelial dysfunction. Erythropoietin (EPO) has emerged as a major tissue-protective cytokine in the setting of stress. We investigated the role of EPO in sepsis-related acute kidney injury (AKI) using a cecal ligation and puncture (CLP) model. Wistar rats were divided into three groups: control (sham-operated); CLP-only; and CLP+EPO. The EPO (4000 IU/kg BW, i.p.) was administered 24 h and 1 h before CLP. To study the early and late effects of EPO on sepsis-induced AKI, we performed experiments at 24 h and 48 h after CLP/sham operation, and we plotted the survival curves. At post-procedure hour 24, CLP+EPO rats presented significantly higher inulin clearance than did CLP-only rats; EPO treatment restored hematocrit levels, as well as mean arterial pressure and metabolic balance. At post-procedure hour 48, CLP+EPO rats presented significantly higher creatinine clearance than did CLP-only rats; EPO treatment restored eNOS levels, suppressed macrophage infiltration, and inhibited NF-B activation,thereby increasing survival. In conclusion, EPO protects renal function and increases survival in this model of sepsis-induced AKI. This protection is dependent on eNOS activation and is partly due to inhibition of the inflammatory response via downregulation of NF- B

<p>Intrauterusna restrikcija rasta (IUGR) se odnosi na stanje u kojem fetus nije u mogućnosti da ostvari svoj genetski potencijal za rast. IUGR je ozbiljan klinički problem i nedavno je povezan sa bolestima odraslog doba kao &scaron;to su hipertenzija, insulin nezavistan diabetes melitus, dislipidemije i ishemijske bolesti srca. Eritropoetin je glavni regulator proliferacije i diferencijacije eritroidnih progenitorskih ćelija zahvaljujući svojoj antiapoptotičkoj aktivnosti. Cilj istraživanja je bio da se ispita uticaj IUGR na bubrege, kao i uticaj eritropotina na bubrege sa IUGR. Eksperimentalna studija je sprovedena u gajili&scaron;tu Pasterovog zavoda u Novom Sadu na 60 mi&scaron;eva rase NMRI. IUGR je izazivana aplikacijom deksametazona gravidnim ženkama. Po rođenju su mladunci bili podeljeni u sedam grupa. Mladuncima je u 1. i 7. danu života davan darbepoetin alfa (DA) u dozama od 1, 4 i 10&mu;g/kg. Dve grupe su predstavljale potomke majki koje su tokom trudnoće dobile DA. Nakon 4 nedelje su uzimani uzorci bubrega i vr&scaron;ena je morfolo&scaron;ka i stereolo&scaron;ka analiza glomerula. Aplikacija deksametazona (100 &mu;g/kg) trudnim mi&scaron;icama dovodi do potomstva sa IUGR. Primena DA kod novorođenih mi&scaron;eva sa IUGR dovodi do bržeg porasta telesne mase u prvih 7 dana života (&bdquo;catch-up― rasta). Mi&scaron;evi rođeni sa IUGR imaju manju povr&scaron;inu glomerula bubrega. Primena DA nakon rođenja i u 7. danu života (4 i 10 &mu;g/kg) kod novorođenih mi&scaron;eva sa IUGR dovodi do hipertrofije glomerula bubrega. IUGR nema uticaja na broj glomerula bubrega mi&scaron;eva. Primena DA nema uticaja na broj glomerula bubrega mi&scaron;eva. Mi&scaron;evi rođeni sa IUGR imaju manju debljinu korteksa bubrega. Primena DA (4 i 10 &mu;g/kg) kod mi&scaron;eva rođenih sa IUGR dovodi do povećanja debljine korteksa bubrega. Davanje DA kod IUGR značajno povećava povr&scaron;inu glomerula i debljinu korteksa bubrega.</p> / <p>Intrauterine growth restriction (IUGR) refers to a condition in which a foetus is not able to achieve its genetic potential for growth. IUGR is a serious clinical problem, and has recently been linked with diseases of adulthood, such as hypertension, insulin-independent diabetes mellitus, dyslipidemia, and ischemic heart disease. Erythropoietin is the major regulator of proliferation and differentiation of erythroid progenitor cells, thanks to its anti-apoptotic activities. The aim of this study was to investigate the effect of IUGR on the kidneys, and the impact of erythropoietin on the kidneys with IUGR. The experimental study was conducted in Pasteur Institute of Novi Sad on 60 mice of NMRI race. IUGR has been imposed with the application of dexamethasone to pregnant females. After birth, the pups were divided into seven groups. DA was administered to the pups on the 1st and 7th day of life (dose 1, 4 and 10 &mu;g/kg). Two groups represented the offspring of the mothers who during pregnancy received DA. After 4 weeks, kidney samples were taken and morphological and stereological analysis of the glomeruli was performed. The application of dexamethasone (100 &mu;g/kg) to pregnant mice leads to their offspring with IUGR. Application of DA to newborn mice with IUGR leads to faster weight gain in the first 7 days of life (&quot;catch-up&quot; growth). Mice born with IUGR have a reduced glomerular surface. Application of DA after birth and on the 7th day of life (4 and 10 &mu;g/kg) in mice with IUGR leads to hypertrophy of the kidney glomeruli. IUGR has no effect on the number of kidney glomeruli. Application of DA has no effect on the number of kidney glomeruli. Mice born with IUGR have a reduced cortical thickness. Application of DA (4 and 10 &mu;g/kg) in mice born with IUGR leads to increased thickness of the kidney cortex. Application of DA to mice with IUGR significantly increases the surface area of the kidney glomeruli and cortical thickness.</p>

Retinopathy of prematurity, a disease characterised by aberrant retinal vascular development in premature neonates, is a leading cause of blindness and visual impairment in childhood. This work sought to examine differences in the susceptibility of inbred rat strains to oxygen-induced retinopathy, a model of human retinopathy of prematurity. The overriding aim was to identify genetic factors in rats that might be generalisable to humans. Newborn rats of six different strains were exposed to alternating cycles of hyperoxia and relative hypoxia for fourteen days. Rats were removed to room air and killed for analysis immediately, to assess oxygen-induced retinal vascular attenuation, or four days later to evaluate the extent of hypoxia-induced vasoproliferation. Whole flat-mounted retinae were stained with fluorophore conjugated isolectin GS-IB4, and measurement of vascular area was conducted using fluorescence microscopy and video-image analysis. A hierarchy of susceptibility to the inhibitory effects of cyclic hyperoxia and relative hypoxia on postnatal retinal vascularization was identified for the rat strains studied. Susceptibility to vascular attenuation was predictive of the subsequent risk of vascular morphological abnormalities. Cross-breeding experiments between susceptible and resistant strains demonstrated that the susceptible phenotype was dominantly inherited in an autosomal fashion. These studies confirmed an association between ocular pigmentation and retinopathy risk, however the finding of differential susceptibility amongst albino rat strains implicated factors in addition to those associated with ocular pigmentation. Quantitative real-time reverse transcription-polymerase chain reaction was used to compare the retinal expression of angiogenic factor genes in susceptible and resistant strains with the aim of identifying a genetic basis for the strain difference. Eight angiogenic factor genes were selected for study: vascular endothelial growth factor (VEGF); VEGF receptor 2; angiopoietin 2; Tie2; pigment epithelium-derived factor; erythropoietin; cyclooxygenase-2 and insulin-like growth factor-1. The most notable difference between strains was the expression of vascular endothelial growth factor (VEGF) during the cyclic hyperoxia exposure period - higher VEGF expression was associated with relative resistance to retinopathy. Other differences in retinal angiogenic factor gene expression between strains, such as higher expression of VEGF receptor 2 and angiopoietin 2 in resistant strains, appeared to be secondary to those in VEGF. Following cyclic hyperoxia, the expression pattern of angiogenic factor genes changed - messenger RNA levels of hypoxia-induced genes, including VEGF, VEGF receptor 2, angiopoietin 2 and erythropoietin, were significantly higher in those strains with larger avascular areas, than in those strains that were relatively resistant to retinopathy. These findings provide firm evidence for hereditary risk factors for oxygen-induced retinopathy in the rat. Differences in the regulatory effects of oxygen on VEGF expression appear to be central to the risk of retinopathy. The potential relevance of these hereditary factors is discussed in the context of the human disease.

Traumatic spinal cord injury (SCI) is a devastating condition for which there is no effective clinical treatment. Neuroprotective molecules that minimize tissue loss have shown promising results; however systemic delivery may limit in vivo benefits due to short systemic half-life and minimal passage across the blood-spinal cord barrier. To overcome these limitations, an injectable intrathecal delivery vehicle comprised of hyaluronan and methylcellulose (HAMC) was developed, and previously demonstrated to be safe and biocompatible intrathecally. Here, HAMC was determined to persist in the intrathecal space for between 4-7 d in vivo, indicating it as an optimal delivery system for neuroprotective agents to reduce tissue degeneration after SCI. HAMC was then investigated as an in vivo delivery system for two neuroprotective proteins: erythropoietin (EPO) and fibroblast growth factor 2 (FGF2). Both proteins demonstrated a diffusive release profile in vitro and maintained significant bioactivity during release. When EPO was delivered intrathecally with HAMC to the injured spinal cord, reduced cavitation in the tissue and significantly improved neuron counts were observed relative to the conventional delivery strategies of intraperitoneal and intrathecal bolus. When FGF2 was delivered intrathecally from HAMC, therapeutic concentrations penetrated into the injured spinal cord tissue for up to 6 h. Poly(ethylene glycol) modification of FGF2 significantly increased the amount of protein that diffused into the tissue when delivered similarly. Because FGF2 is a known angiogenic agent, dynamic computed tomography was developed for small animal serial assessment of spinal cord hemodynamics. Following SCI and treatment with FGF2 from HAMC, moderate improvement of spinal cord blood flow and a reduction in permeability were observed up to 7 d post-injury, suggesting that early delivery of neuroprotective agents can have lasting effects on tissue recovery. Importantly, the entirety of this work demonstrates that HAMC is an effective short-term delivery system for neuroprotective agents by improving tissue outcomes following traumatic SCI.