The role of food restriction on CCl4-induced cirrhosis model in rats.

Effects of food restriction on susceptibility to the toxic effect of some chemicals are controversial. In order to identify an exposure model that could maximize cirrhosis and minimize mortality rate, this study aimed to evaluate the effect of food restriction on tetrachloride carbon (CCl(4))-induced cirrhosis model in rats. Fifty-three male Wistar rats received CCl(4) 0.25 ml/kg weekly intragastrically once a week. Thirty-three had 44% food restriction (group 1); 10 rats had 25% food restriction (group 2); and 10 rats received ad libitum food (group 3). After 10 weeks, the animals were sacrificed and liver sections were collected for histology. Of the 53 animals enrolled for the study, 22 (41.5%) died before completing 10-week CCl(4). Mortality rate was significantly higher in group 1 compared to other groups (p<0.05). Cirrhosis was significantly more prevalent in group 1 than in group 3 (p<0.01), but without significant difference between groups 1 and 2 (p=0.624). We concluded that food restriction is an important issue to be considered when establishing a CCl(4)-induced cirrhosis model in rats. Moreover, there is an ideal range of food intake that predisposes to liver damage without increasing mortality leading to a more effective model.

White matter lesions in Fabry disease before and after enzyme replacement therapy: a 2-year follow-up.

PURPOSE: To report the clinical and neuroimaging, central nervous system (CNS) findings of patients with Fabry disease (FD) during 24 months of enzyme replacement therapy (ERT) with agalsidase-alpha. METHOD: Eight patients were included. Six completed 24 months of ERT. Clinical and magnetic resonance imaging (MRI) data were obtained at 0, 12 and 24 months of ERT. White matter lesions (WML) were evaluated as well as their relation to age, symptoms and neurological examination (CNS score). RESULTS: MRI was stable in 3 patients. WML and CNS score worsened in one patient, fluctuated in another, and improved in the sixth patient. In the whole series, there were 15 WML at baseline, and 19 at the 24th month. In two years, 4 lesions disappeared, whereas 8 appeared. CONCLUSION: A widespread pattern of silent WML in FD was seen. In two years, some WML appeared, and some disappeared. If these phenomena were related to the natural history, remains to be demonstrated.

Replication Study of Polymorphisms Associated With Brain Arteriovenous Malformation in a Population From South of Brazil.

INTRODUCTION: The aim of this study was to reproducibly determine if any of the polymorphisms were associated with the susceptibility to brain arteriovenous malformations (BAVM) or the risk of intracranial hemorrhage (ICH) presentation. METHODS: We recruited 63 BAVM patients and 96 controls. The polymorphisms selected for evaluation were apolipoprotein E (APOE), tumor necrosis factor alpha (TNF 238G>A - rs361525), interleukin 1 beta (IL1B 511C>T - rs16944 and IL1B -31T>C - rs1143627), activin-like kinase 1 (ACVRL1 IVS3-35A>G - rs2071219), endoglin (ENG 207G>A - rs11545664), and interleukin 6 (IL6 174G>C - rs1800795). RESULTS: In the single analysis, we observed statistically significant differences in the allele distributions for IL1B -31T>C (rs1143627) between the BAVM patients and control subjects (P = 0.02). There was a trend toward significance for the association between the IL1B 511C>T (rs16944) allele and BAVM risk (P = 0.07). In further logistic regression analysis, no polymorphism was significantly associated with the risk of BAVM. No polymorphisms were associated with hemorrhage presentation according to both single and multivariable analyses. CONCLUSIONS: In our sample from a south Brazil population, we found no association between the risks of BAVM and ICH presentation with any of the selected polymorphisms.

Exercise on Progenitor Cells in Healthy Subjects and Patients with Type 1 Diabetes.

PURPOSE: To evaluate the acute effect of aerobic exercise (AE) and resistance exercise (RE) on the release of endothelial progenitor cell (EPCs, CD34+/KDR+/CD45 dim) and vascular function in type 1 diabetes (T1DM). METHODS: Fourteen men with T1DM and 5 nondiabetic controls were randomly assigned to 40-min AE (60% VO 2peak) and RE sessions (60% 1-RM). The study had a crossover design, and interventions were 1 wk apart. Venous occlusion plethysmography (blood flow, reactive hyperemia, and vascular resistance) and blood collection (EPC levels, flow cytometry) were done immediately before and after exercise sessions. RESULTS: Patients were 30.3 ± 1.6 yr-old, HbA1c 7.7% ± 0.2%; controls were 26.8 ± 2.3 yr-old. Groups did not differ in EPC levels at baseline or in relation to exercise. Over time, exercise did not induce changes in patients with T1DM, whereas, in controls, EPCs were decreased after AE (-10.7%, P = 0.017) and increased after RE (+12.2%, P = 0.004). Compared with baseline, blood flow increased and vascular resistance decreased after RE in both groups. Reactive hyperemia was increased 10 min after AE and RE sessions in patients with T1DM (36.5% and 42.0%, respectively) and in controls (35.4% and 74.3%), but no group differences were observed between groups in response to exercise. CONCLUSIONS: Despite the increased vascular reactivity in both groups after both exercise sessions, EPCs were only influenced by exercise in controls. The unchanged number of EPCs in T1DM after exercise sessions might indicate a blunted endothelium regenerating capacity, revealing an early deterioration of the functional arterial characteristics not disclosed by only evaluating vascular functional variables.

Association between plasma adiponectin and arteriolar vessel caliber among elderly hypertensive subjects.

Circulating adiponectin has been related to vascular diseases, but few studies examined the relationship between plasma adiponectin and microvascular abnormalities among hypertensive individuals. We tested the association between plasma adiponectin level and retinal vessel calibers in patients with hypertension.This study included 172 patients with confirmed hypertension, aged 18-80 years. Subjects with recent cardiovascular events, advanced heart failure and end-stage renal disease were excluded. Arteriolar and venular calibers were measured in retinographies using a microdensitometric image-processing method. Blood pressure was measured using a validated oscillometric device. We observed a statistically significant inverse association between plasma adiponectin and arteriolar caliber among participants aged 60 years or older after controlling for confounders (Adjusted ß = -0.42; P = .001). In the final model, HbA1C and low-density lipoprotein also remained independently associated with arteriolar caliber. There was no association of adiponectin with venular caliber and retinal vessel calibers in participants <60 years old.Adiponectin is inversely associated with retinal arteriolar caliber in elderly hypertensive participants, suggesting that plasma adiponectin may be a marker of microvascular damage and of higher cardiovascular risk in this age stratum.

Protective effects of melatonin-loaded lipid-core nanocapsules on paraquat-induced cytotoxicity and genotoxicity in a pulmonary cell line.

Many acute poisonings lack effective and specific antidotes. Due to both intentional and accidental exposures, paraquat (PQ) causes thousands of deaths annually, especially by pulmonary fibrosis. Melatonin (Mel), when incorporated into lipid-core nanocapsules (Mel-LNC), has enhanced antioxidant properties. The effects of such a formulation have not yet been studied with respect to mitigation of PQ- induced cytotoxicity and DNA damage. Here, we have tested whether Mel-LNC can ameliorate PQ-induced toxicity in the A549 alveolar epithelial cell line. Physicochemical characterization of the formulations was performed. Cellular uptake was measured using nanocapsules marked with rhodamine B. Cell viability was determined by the MTT assay and DNA damage was assessed by the comet assay. The enzyme-modified comet assay with endonuclease III (Endo III) and formamidopyrimidine glycosylase (FPG) were used to investigate oxidative DNA damage. Incubation with culture medium for 24h did not alter the granulometric profile of Mel-LNC formulations. Following treatment (3 and 24h), red fluorescence was detected around the cell nucleus, indicating internalization of the formulation. Melatonin solution (Mel), Mel-LNC, and LNC did not have significant effects on cell viability or DNA damage. Pre-treatment with Mel-LNC enhanced cell viability and showed a remarkable reduction in % DNA in tail compared to the PQ group; this was not observed in cells pre-treated with Mel. PQ induces oxidative DNA damage detected with the enzyme-modified comet assay. Mel-LNC reduced this damage more effectively than did Mel. In summary, Mel-LNC is better than Mel at protecting A549 cells from the cytotoxic and genotoxic effects of PQ.

Exon 11 mutations, Ki67, and p16(INK4A) as predictors of prognosis in patients with GIST.

Prognostic biomarkers for GIST are under investigation. The aim of this study was to assess whether exon 11 mutations, Ki67, and p16(INK4A) are predictors of prognosis in GIST. Consecutive GIST cases (n=84) had their specimens evaluated for exon 11 mutations and expression of Ki67 and p16(INK4A). Surgical cases were categorized according to NIH and Miettinen's classification, and survival was analyzed from hospital database. GISTs were predominately gastric (45%) and with spindle cell morphology (74%). The risk category was very low or low in 28%, intermediate in 23%, and high in 49%. Exon 11 mutation was identified in 29 (48%) out of 60 cases studied. There were 12 point mutations, 10 deletions, 4 duplications, and 3 double mutations. A third of GISTs had either high Ki67 index (>3%) or negativity for p16(INK4A). In multivariate analysis, independent predictors of mortality were Ki67>3% (HR=7.3; P=0.036) and high mitotic index (HR=10.4; P=0.043). There was no association between exon 11 mutations and survival. This study suggests that Ki67>3% is an independent predictor of poor prognosis in patients with GIST. Exon 11 mutations and negativity for p16(INK4A) need further studies to address the prognostic value.

Seminiferous epithelium of rats with food restriction and carbon tetrachloride-induced cirrhosis.

OBJECTIVE: Analyze the changes in the seminiferous epithelium in rats with carbon tetrachloride-induced cirrhosis (CCl4). MATERIALS AND METHODS: Forty-eight male Wistar rats aged 45-50 days, weighing 150-180 grams were used. Twenty-two rats underwent CCl4-induced cirrhosis with CCl4 0.25 mL/Kg weekly intragastrically once a week, during 10 weeks. Additionally, they had a 44% food restriction diet (Group 1). The control group was divided in two subgroups: 13 rats had a 44% food restriction diet and no CCl4 (Group 2) and 10 rats were not submitted to CCl4 or food restriction (Group 3). After 10 weeks, the rats were sacrificed and liver sections were collected for histological analysis. The testicular analysis was carried out to evaluate the frequency of tubules in stages VIII and XIV. RESULTS: The mean rates of stage VIII in animals with food restriction plus CCl4-induced cirrhosis and food restriction without CCl4 were significantly different from animals without either food restriction or CCl4 (18.1 +/- 5.5%, 20.5 +/- 2.5% and 13.4 +/- 3.5%, respectively, p = 0.002). The mean rate of stage VIII in rats with cirrhosis was not significantly different from rats without cirrhosis (18.1 +/- 5.5% and 17.4 +/- 4.6% respectively). The mean frequency of stage XIV in rats with cirrhosis was significantly greater than rats without cirrhosis (4.7 +/- 2.3% and 6.8 +/- 1.9% respectively, p = 0.027). CONCLUSION: Animals with CCl4-induced cirrhosis and food restriction have shown alterations in spermatogenic cycle that were not seen in rats without CCl4-induced cirrhosis and food restriction.

Seminiferous epithelium of rats with food restriction and carbon tetrachloride-induced cirrhosis

OBJECTIVE: Analyze the changes in the seminiferous epithelium in rats with carbon tetrachloride-induced cirrhosis (CCl4). MATERIALS AND METHODS: Forty-eight male Wistar rats aged 45-50 days, weighing 150-180 grams were used. Twenty-two rats underwent CCl4-induced cirrhosis with CCl4 0.25 mL/Kg weekly intragastrically once a week, during 10 weeks. Additionally, they had a 44 percent food restriction diet (Group 1). The control group was divided in two subgroups: 13 rats had a 44 percent food restriction diet and no CCl4 (Group 2) and 10 rats were not submitted to CCl4 or food restriction (Group 3). After 10 weeks, the rats were sacrificed and liver sections were collected for histological analysis. The testicular analysis was carried out to evaluate the frequency of tubules in stages VIII and XIV. RESULTS: The mean rates of stage VIII in animals with food restriction plus CCl4-induced cirrhosis and food restriction without CCl4 were significantly different from animals without either food restriction or CCl4 (18.1 ± 5.5 percent, 20.5 ± 2.5 percent and 13.4 ± 3.5 percent, respectively, p = 0.002). The mean rate of stage VIII in rats with cirrhosis was not significantly different from rats without cirrhosis (18.1 ± 5.5 percent and 17.4 ± 4.6 percent respectively). The mean frequency of stage XIV in rats with cirrhosis was significantly greater than rats without cirrhosis (4.7 ± 2.3 percent and 6.8 ± 1.9 percent respectively, p = 0.027). CONCLUSION: Animals with CCl4-induced cirrhosis and food restriction have shown alterations in spermatogenic cycle that were not seen in rats without CCl4-induced cirrhosis and food restriction.

White matter lesions in Fabry disease before and after enzyme replacement therapy: a 2-year follow-up / Lesões da substância branca na doença de Fabry antes e depois da terapia de reposição enzimática: um seguimento de 2 anos

PURPOSE: To report the clinical and neuroimaging, central nervous system (CNS) findings of patients with Fabry disease (FD) during 24 months of enzyme replacement therapy (ERT) with agalsidase-alpha. METHOD: Eight patients were included. Six completed 24 months of ERT. Clinical and magnetic resonance imaging (MRI) data were obtained at 0, 12 and 24 months of ERT. White matter lesions (WML) were evaluated as well as their relation to age, symptoms and neurological examination (CNS score). RESULTS: MRI was stable in 3 patients. WML and CNS score worsened in one patient, fluctuated in another, and improved in the sixth patient. In the whole series, there were 15 WML at baseline, and 19 at the 24th month. In two years, 4 lesions disappeared, whereas 8 appeared. CONCLUSION: A widespread pattern of silent WML in FD was seen. In two years, some WML appeared, and some disappeared. If these phenomena were related to the natural history, remains to be demonstrated.

OBJETIVO: Relatar os achados neurológicos e de imagem do sistema nervoso central (SNC), observados durante 24 meses de tratamento de reposição enzimática (ERT) com agalsidase-alfa, em pacientes com a doença de Fabry (FD). MÉTODO: 8 pacientes foram incluídos; 6 completaram 24 meses de ERT. Os dados foram obtidos aos 0, 12 e 24 meses de ERT. Lesões de substância branca (WML) foram avaliadas assim como sua relação com a idade e o exame neurológico (escore SNC). RESULTADOS: Os achados de ressonância nuclear magnética foram estáveis em 3 pacientes. As WML e o escore SNC pioraram em um caso; flutuaram em um outro caso; e melhoraram no sexto paciente. No todo, havia 15 WML antes da ERT e 19 WML depois de 24 meses de ERT. Em dois anos, 4 lesões desapareceram e 8 novas surgiram. CONCLUSÕES: Viu-se um padrão difuso de WML assintomáticas, na FD. Em dois anos, algumas WML surgiram, enquanto outras desapareceram. Resta demonstrar se esses fenômenos fazem parte da história natural da doença.