The importance of homocysteine levels in the prognosis of patients with chronic renal disease and in hemodialysis patients / Importância dos níveis de homocisteína no prognóstico de pacientes com doença renal crônica e em pacientes em hemodiálise

ABSTRACT Introduction: Homocysteine (Hcy) is one of the metabolites of methionine (Met), an essential diet-derived amino acid. There is a close relationship between high plasma Hcy levels and declining renal function. Plasma and urinary Hcy level has been the target of studies as a biomarker that forecasts poor outcome in renal patients and in hemodialysis patients. This review evaluates the main studies that sought to correlate Hcy and poor prognosis in renal disease as well as the treatments proposed for the reduction of plasma Hcy levels in these patients. Conclusion: Hcy could be an important biomarker of renal disease progression mainly in hemodialysis patients. We emphasize the importance of normalizing plasma levels of this amino acid to ensure a better prognosis in kidney disease.

RESUMO Introdução: A homocisteína (Hcy) é um dos metabólitos da metionina (Met), um aminoácido essencial proveniente da dieta. Existe uma estreita relação entre os altos níveis plasmáticos de Hcy e o declínio da função renal. A Hcy plasmática e urinária tem sido alvo de estudos como um biomarcador capaz de sinalizar o prognóstico em doentes renais e em pacientes em hemodiálise. Esta revisão avalia os principais estudos que buscaram correlacionar a Hcy e o prognóstico da doença renal e descreve os tratamentos propostos para a redução dos níveis plasmáticos de Hcy nesses pacientes. Conclusão: A Hcy pode ser um biomarcador da progressão na doença renal, principalmente em pacientes hemodialíticos. Ressaltamos a importância da normalização dos níveis plasmáticos desse aminoácido para garantir um melhor prognóstico na doença renal.

Remote ischemic preconditioning in myocardial protection in hemodialysis patients.

BACKGROUND: Remote ischemic preconditioning (RIPC) is a procedure that generates a brief period of ischemia followed by reperfusion. The role of RIPC in protecting myocardial ischemia during hemodialysis is not yet established. The aim of the study was to evaluate RIPC myocardial protection as evaluated by ultrasensitive I troponin in hemodialysis outpatients. PATIENTS AND METHODS: A double-blind randomized trial with two groups: intervention submitted to RIPC and control group without RIPC. Intervention group received RIPC in three consecutive hemodialysis sessions. Blood samples were taken before and after each session. Blood urea nitrogen for calculation of single-pool Kt/v and ultrasensitive I troponin were measured to evaluate dialysis adequacy and myocardial injury. RESULTS: A total of 47 patients were randomized. About 60.8% were men and 54% were diabetic. The mean single-pool Kt/v was 1.51 in the intervention group and 1.49 in control. The ultrasensitive troponin I measured no significant change from the time of collection: before or after dialysis. CONCLUSION: The RIPC applied in three consecutive sessions did not demonstrate superiority to control, therefore another study tested RIPC in 12 consecutive sessions with a positive result in myocardial protection. In our study, more than half of the patients were diabetic. Diabetic patients have a trend to show a lower response to RIPC because of the greater presence of collateral coronary circulation. In summary, in this model there was no interference of RIPC in ultrasensitive troponin I values, but troponin had a high negative predictive value for myocardial infarction in all tested models.

Serum NGAL and Cystatin C Comparison With Urinary Albumin-to-Creatinine Ratio and Inflammatory Biomarkers as Early Predictors of Renal Dysfunction in Patients With Type 2 Diabetes.

INTRODUCTION: Diabetic nephropathy is associated with specific histological changes. Early detection of poor glomerular and tubular function can be achieved with biomarkers of diabetes. The aim of this study was to evaluate the accuracy of kidney dysfunction biomarkers in type 2 diabetes (T2D). METHODS: Patients with T2D were grouped according to their glycated hemoglobin level. Patients' urine and blood samples were taken to measure cystatin C (CysC), neutrophil gelatinase-associated lipocalin, beta-trace protein levels, and the first morning void albumin-to-creatinine ratio. Patients in the end stage of renal disease or receiving dialysis were not included. Receiver operating characteristic curves were generated, and the areas under the curve were compared with the performance of the biomarkers used to evaluate kidney dysfunction in T2D. RESULTS: Ninety patients with T2D were chosen. CysC was positively correlated with creatinine (P < 0.001), estimated glomerular filtration rate (P < 0.001), and urinary beta-trace protein (P = 0.01). The area under the curve was 0.635 for CysC, 0.621 for serum neutrophil gelatinase-associated lipocalin, and 0.660 for the albumin-to-creatinine ratio. A crude logistic regression model showed a positive association between serum CysC (P = 0.01) and serum neutrophil gelatinase-associated lipocalin (P < 0.001). A linear regression model showed a positive association between serum CysC, creatinine, and estimated glomerular filtration rate (P < 0.001) but did not show a positive association with glycated hemoglobin (P = 0.892). DISCUSSION: Neutrophil gelatinase-associated lipocalin and serum CysC were positively associated with the presence of renal dysfunction and had better performance on receiver operating characteristic analysis than the other markers evaluated in patients with T2D without kidney dysfunction.

Quality of Life of Post-Mastectomy Women Living in a Semi-Arid Region of Brazil.

Health is the major reference regarding quality of life; when it comes to breast cancer in particular, the loss of a breast traumatically affects a woman's life, reflecting on her quality of life. Recognizing this problem, our aim was to investigate the quality of life of women who live in a semi-arid region of Brazil after breast cancer mastectomy. In this exploratory, transversal and observational study, a Brazilian variantof the shorter version of the original instrument from the World Health Organization Quality of Life (WHOQOL-BREF), applied in the study population, was analyzed and their socio-demographic profile was obtained. The sample was composed of 50 mastectomized women. The 50 included patients comprised women at a mean age of 54 years. Most of them had finished elementary school, and their average income was one Brazilian minimum monthly wage. Regarding the data related to quality of life, the highest score was found in the social relationships domain (4.29) followed by the psychological (4.09) and environmental (3.88) domains. The lowest score observed was for the physical domain (3.48). With these findings we can say that social and psychological parameters are driving factors of the quality of life in post-mastectomy women. Therefore, these results are useful to establish strategies to improve the quality of life of breast cancer mastectomy patients.

Evaluation of MCT1, MCT4 and CD147 Genes in Peripheral Blood Cells of Breast Cancer Patients and Their Potential Use as Diagnostic and Prognostic Markers.

BACKGROUND: Patients with breast cancer-the deadliest cancer among women-are at constant risk of developing metastasis. Oxidative stress and hypoxia are common feature of tumor cells that can proliferate even in a resultant metabolic acidosis. Despite the low extracellular pH, intracellular pH of tumor cells remains relatively normal, or even more alkaline due to the action of a membrane protein family known as monocarboxylate transporters (MCTs). The objective of this study was to verify the diagnostic and prognostic value of MCT1, MCT4 and CD147 in tumor and peripheral blood samples of patients with breast cancer undergoing chemotherapic treatment. METHODS: Differential expression of MCT1, MCT4 and CD147 obtained by qPCR was determined by 2-ΔΔCq method between biological samples (tumor and serial samples of peripheral) of patients (n = 125) and healthy women (n = 25). RESULTS: tumor samples with higher histological grades have shown higher expression of these markers; this higher expression was also observed in blood samples obtained at diagnosis of patients when compared to healthy women and in patients with positive progression of the disease (metastasis development). CONCLUSION: markers studied here could be a promising strategy in routine laboratory evaluations as breast cancer diagnosis and prognosis.

Acne in adult women and the markers of peripheral 3 alpha-diol G activity.

BACKGROUND: Acne in adult women is a frequent hard-to-manage disease with many relapse cases. It mostly interferes with the quality of life of patients, bringing them major metabolic and social losses. As androgenic hormones play a very important role in the acne pathogenesis, the early diagnosis of hyperandrogenic states is very useful for the proper evaluation of each patient and for a better choice of therapeutic management. Defining a pattern for laboratory profile analysis is important for the control of relapses of acne breakouts in adult women, which lately has been the aim of many published studies. AIM: To establish the relation between 3 alpha-diol G levels and acne in female patients with normal androgenic status without menstrual dysfunctions. PATIENTS/METHODS: The evaluation of serum 3 alpha-androstanediol glucuronide levels through an enzymatic immunoassay method (Androstanediol Glucuronide ELISA Kit) for a direct quantitative measurement in 26 patients with grade II and III acne, ages ranging from 13 to 50. RESULTS: Among the analyzed patients, 83% had grade II acne, and among this total, 60% were aged 14 or over. According to age, 12 studied patients showed serum 3 alpha-diol G levels within normal range and 11 patients had increased levels. CONCLUSIONS: A total of 60% of adult women with acne present increased levels of androgens and among those with normal levels and without menstrual dysfunctions, 50% show an increase in 3 alpha-diol G. Therefore, a pharmacological approach with anti-androgenic drugs for acne therapy in most of these patients is advisable.

Plasma levels of E-cadherin and MMP-13 in prostate cancer patients: correlation with PSA, testosterone and pathological parameters.

AIMS AND BACKGROUND: Prostate cancer is the most common malignant tumor in men. Serum prostate-specific antigen (PSA), Gleason score and clinical range at the time of diagnosis are important factors to predict prognosis and outcome after therapy but additional accurate and reliable biomarkers are still wanted. So far, few biomarkers of prostate cancer have been successfully implemented and are being used in clinical practice. However, modifications of E-cadherin and MMP-13 expression are likely to be involved in prostate cancer invasion and thus are potential biomarkers for prognosis. PATIENTS AND METHODS: We analyzed the concentrations of E-cadherin and MMP-13 in plasma of patients with prostate cancer at diagnosis and 3 and 6 months after the beginning of treatment and related these measures to free and total PSA and other clinical features. RESULTS: The concentration of E-cadherin was lower in patients with prostate cancer compared to the control group, but there was no difference in the concentration of MMP-13 between these two groups. During treatment, however, we found no significant differences between the concentrations of MMP-13 and E-cadherin, but we observed a significant positive correlation between total PSA and E-cadherin plasma concentration at the third month of treatment and between total testosterone and MMP-13 plasma concentration before the start of treatment. CONCLUSIONS: The results suggest that these parameters could be used both in the diagnosis and prognosis of prostate cancer.

Inability of Turbidimetry Method in Detecting Glycated Hemoglobin to Select Diabetes Mellitus Patients According to Their Concentrations of Blood Glucose Levels.

BACKGROUND: Diabetes mellitus is a chronicle illness in which there is a high blood glucose level defined as hyperglycemia, resulted by a deficiency in insulin secretion and/or in its action. Nowadays, it is being seen as a public health problem and is reaching increasing proportions with regard to the appearance of new cases. For diagnosis, sensible and accurate methods should be used to avoid complications of the sickness. The measure of glycated hemoglobin may not be used for diagnosis, but is the reference method to evaluate the grade of glycemic control in the long term, reflecting the blood glucose level in the latest 2-3 months. The aim of this study was to evaluate the grade of concordance between turbidimetry and liquid chromatography methods in the glycated hemoglobin determination and to estimate the sensibility and specificity values of turbidimetry. METHODS: This study included 133 blood samples obtained from patients and healthy donors, ageing between 18 and 80 years with glycemic values between 58 and 473 mg/dl. RESULTS AND CONCLUSION: Turbidimetry is a useful method for determining glycemic levels above 100 and over 200 mg/dl, but does not have the ability to select samples with intermediary blood glucose concentrations.

Systemic chemotherapy interferes in homocysteine metabolism in breast cancer patients.

BACKGROUND: Hyperhomocysteinemia in breast cancer (BC) patients can be a risk factor for thromboembolic events. This study aimed to evaluate homocysteine and its cofators (folic acid and vitamin B12) concentrations and platelet count at diagnosis of BC, 3 and 6 months after the beginning of chemotherapy treatment and to correlate them with clinical data. METHODS: Thirty-five BC patients were included; blood samples were obtained by venipuncture. Plasmatic Hcy and cofactors concentrations were measured by competitive chemiluminescent enzyme immunoassay method. Platelet count was done using an automated analyzer. Statistical analysis was performed using the software SPSS. RESULTS: During chemotherapy, homocysteine (P = 0.032) and vitamin B12 (P < 0.001) concentrations increased, while folate and platelets decreased (P < 0.001). Among the clinical data, the menopausal status showed significant positive correlation (P = 0.022) with homocysteine concentration increase. CONCLUSIONS: Evaluation of homocysteine concentrations during chemotherapy is extremely important because their levels increase during chemotherapy treatment, thus increasing the risk of thromboembolism development.

Endothelial, renal and hepatic variables in Wistar rats treated with Vancomycin.

Vancomycin (VCM) is indicated in combat against Gram-positive infections, but it is not considered a first-choice drug because of its adverse effects. It is believed that oxidative stress is the primary mechanism of endothelial injury and the consequent VCM toxicity, which varies from phlebitis to nephrotoxicity. Moreover, dose recommendations, dilution, rates and types of infusion are still controversial. The aim of this study was to determine the effect of different VCM dilutions in endothelial, liver and kidney injuries by biochemical parameters and histopathological analysis. Wistar rats were randomly divided into six groups and subjected to femoral vein cannulation for drug administration. Control groups received 0.9 ml of saline and the others received VCM (10mg/Kg/day) at dilutions of 5.0 and 10.0 mg/mL for 3 and 7 days. Homocysteine, hs-CRP, AST, ALT, GGT, urea, creatinine, lycopene, alpha-tocopherol, beta-carotene and retinol were analyzed. Kidney, liver and cannulated femoral vein fragments were collected.This study showed alterations in ALT which featured hepatotoxicity. However, drug dilutions were not able to show changes in other biochemical parameters. In contrast, kidney and endothelium pathological changes were observed. More studies are needed to characterize VCM induced kidney and endothelium toxicity and biochemical markers able to show such morphological modifications.

Standardization of a method to study angiogenesis in a mouse model.

In the adult organism, angiogenesis is restricted to a few physiological conditions. On the other hand, uncontrolled angiogenesis have often been associated to angiogenesis-dependent pathologies. A variety of animal models have been described to provide more quantitative analysis of in vivo angiogenesis and to characterize pro- and antiangiogenic molecules. However, it is still necessary to establish a quantitative, reproducible and specific method for studies of angiogenesis factors and inhibitors. This work aimed to standardize a method for the study of angiogenesis and to investigate the effects of thalidomide on angiogenesis. Sponges of 0.5 x 0.5 x 0.5 cm were implanted in the back of mice groups, control and experimental (thalidomide 200 mg/K/day by gavage). After seven days, the sponges were removed. The dosage of hemoglobin in sponge and in circulation was performed and the ratio between the values was tested using nonparametric Mann-Whitney test. Results have shown that sponge-induced angiogenesis quantitated by ratio between hemoglobin content in serum and in sponge is a helpful model for in vivo studies on angiogenesis. Moreover, it was observed that sponge-induced angiogenesis can be suppressed by thalidomide, corroborating to the validity of the standardized method.

Incidence of Leptospirosis infection in the East Zone of Sao Paulo City, Brazil.

BACKGROUND: Leptospirosis is a zoonosis which is spread through contamined running water. This contaminations is seriously affected by the flooding which occurs in the area surrounding the Aricanduva river. The transmission of the disease results mainly from the contact of water with soil contaminated by the urine of infected animals. We aimed to conduct an epidemiological survey on Leptospirosis cases in Sao Paulo East Zone area. METHOD: The analysis conducted in this study was based on data collected from the health authorities of that region close the Aricanduva river between 2007 and 2008 years, which give the rates of confirmed cases, mortality and death from human Leptospirosis. Other information concerned with the relationships among rainfall index, points of flooding and incidence of Leptospirosis. RESULTS: We observed a direct and important water contamination. Records of flooding points and dates of the reported cases in the region showed a direct relationship from which the period of higher rainfall also recorded an increase in cases. The annual record of the city and the region and rainfall regions also presented correlation. CONCLUSION: The association between the indices of flooding and Leptospirosis cases indicates that preventive measures are necessary to avoid exposing the community.

Circulating tumor cell detection during chemotherapy in patients with breast cancer is not associated with plasma homocysteine levels.

Breast cancer remains the second most frequent type of cancer in the world and the first among women, and systemic chemotherapy is an adjuvant therapeutic modality that improves survival in a great part of patients. Women with breast cancer, however, frequently show a higher risk of thromboembolism, an event associated to hyperhomocysteinemia and the presence of circulating tumor cells (CTC). Our aim is to correlate the presence of CTCs, detected by the analysis of CK19 and c-erbB2 gene expressions, and the homocysteine plasma levels in the peripheral blood in patients with breast cancer undergoing chemotherapy. Epithelial marker expression (CK19 and c-erbB2) and homocysteine levels were analyzed in a mononuclear fraction of the peripheral blood and plasma, respectively, obtained from 35 patients diagnosed with breast cancer at diagnosis and throughout chemotherapy treatment. No significant relation between the CK19 and c-erbB2 expressions and hyperhomocysteinemia was observed at any moment of the evaluation throughout the chemotherapy treatment (3 and 6 months after the onset). Among clinical data, only menopausal status showed a statistically significant correlation with homocysteine concentration. Although differences in the expressions of the analyzed epithelial markers were detected at 3 and 6 months of chemotherapy treatment, no relation between plasma homocysteine variations and the CK19 and c-erbB2 gene expressions was found in patients under chemotherapy treatment at any moment of the evaluation, suggesting that chemotherapy affects the expressions of the studied genes independently.

Lack of reliability of nanotechnology in the of free plasma DNA in samples of patients with prostate cancer.

BACKGROUND: Several studies seek biological markers that give diagnostic and degree of tumor development. The aim of this study was to validate the determination of plasma DNA using nanotechnology (Nanovue™-NV) in samples of 80 patients with prostate cancer. METHODS: Blood samples of 80 patients of the Urology Ambulatory of Faculdade de Medicina do ABC with prostate cancer confirmed by anatomical-pathology criteria were analyzed. DNA extraction was performed using a GFX TM kit (Amersham Pharmacia Biotech, Inc, USA) following the adapted protocol. Plasma was subjected to centrifugation. RESULTS: There was a big difference between the first and the second value obtained by NanoVue Only two samples had no differences between duplicates. Maximum difference between duplicates was 38 µg/mL. Average variation between 51 samples was 10.29 µg/mL, although 21 samples had differences above this average. No correlation was observed between pDNA obtained by traditional spectrophotometry and by nanotechnology. CONCLUSION: Determination of plasma DNA by nanotechnology was not reproducible.

Mild systemic oxidative stress in the subclinical stage of Alzheimer's disease.

Alzheimer's disease (AD) is a late-onset, progressive degenerative disorder that affects mainly the judgment, emotional stability, and memory domains. AD is the outcome of a complex interaction among several factors which are not fully understood yet; nevertheless, it is clear that oxidative stress and inflammatory pathways are among these factors. 65 elderly subjects (42 cognitively intact and 23 with probable Alzheimer's disease) were selected for this study. We evaluated erythrocyte activities of superoxide dismutase, catalase, and glutathione peroxidase as well as plasma levels of total glutathione, α-tocopherol, ß-carotene, lycopene, and coenzyme Q10. These antioxidant parameters were confronted with plasmatic levels of protein and lipid oxidation products. Additionally, we measured basal expression of monocyte HLA-DR and CD-11b, as well as monocyte production of cytokines IL1-α, IL-6, and TNF-α. AD patients presented lower plasmatic levels of α-tocopherol when compared to control ones and also higher basal monocyte HLA-DR expression associated with higher IL-1α production when stimulated by LPS. These findings support the inflammatory theory of AD and point out that this disease is associated with a higher basal activation of circulating monocytes that may be a result of α-tocopherol stock depletion.

Effects of ethanol on CYP2E1 levels and related oxidative stress using a standard balanced diet.

Expression of cytochrome P4502E1 (CYP2E1) is very much influenced by nutritional factors, especially carbohydrate consumption, and various results concerning the expression of CYP2E1 were obtained with a low-carbohydrate diet. This study describes the effects of ethanol treatment on CYP2E1 levels and its relationship with oxidative stress using a balanced standard diet to avoid low or high carbohydrate consumption. Rats were fed for 1, 2, 3, or 4 weeks a commercial diet plus an ethanol-sucrose solution. The results have shown that ethanol administration was associated with CYP2E1 induction and stabilization without related oxidative stress. Our findings suggest that experimental models with a low-carbohydrate/high-fat diet produce some undesirable CYP2E1 changes that are not present when a balanced standard diet is given.

Adhesion molecules affected by treatment of lung cancer cells with epidermal growth factor.

Lung cancer is one of the leading causes of death in the world. Some tumor events are attributed to an important group of molecules (cadherins and integrins). We evaluated the interactions of cell adhesion molecules in cell lines from lung cancer. Two lung cancer cell lines were nonmetastatic (H358 and H441) and two were metastatic (H1299 and H292). All cell lines were treated with epidermal growth factor (EGF), and Western blot analysis was performed to assess the interactions between these proteins. The bronchoalveolar cells H358 showed the three analyzed proteins: E-cadherin, ß-catenin, and p120 catenin. The adenocarcinoma cells H441 did not present p120 catenin, and carcinoma cells did not show E-cadherin (H1299) or p120 catenin (H292). FAK (pTyr925) was dephosphorylated in adenocarcinoma cells H441, absent in carcinoma cells H1299, and upregulated in the other carcinoma cells H292. p130Cas showed no difference when the cell lines were treated with EGF for 30 min; it was absent in the metastatic carcinoma cells H1299. Paxillin was dephosphorylated in adenocarcinoma cells H441 and also absent in other metastatic carcinoma cells H292. Vinculin showed the same results, and talin was downregulated in adenocarcinoma cells H441 when the cells were treated with EGF. Rap1 was downregulated and PYK2 was upregulated in the same cell line. Our data help to comprehend the mechanism involved in cell migration to the blood and metastasis generation. In conclusion, the expression patterns of cell-cell adhesion were not affected by EGF treatment but it affected cell-extracellular matrix adhesion.

The management of menopause with complementary and alternative medicine using an experimental model: ovariectomized rats.

AIM: The purpose of this research was to assess the effects of the flower essences She Oak and Bush Fuchsia on behavioral anxiety in ovariectomized (OVX) rats. METHODS: For four weeks, OVX rats received the flower essences She Oak, Bush Fuchsia or a combination of the two. After flower therapy, the animals were subjected to an elevated plus maze (EPM) behavioral anxiety-test. Cortisol blood level was also evaluated. RESULTS: OVX rats treated with the flower essence She Oak became less anxious and had more entries in the EPM open arms. On the other hand, OVX rats treated with the Bush Fuchsia essence spent more time in the EPM closed arms. This finding is similar to those obtained with controls. In addition, OVX rats that received She Oak and Bush Fuchsia in combination presented the same results as those receiving the Bush Fuchsia alone. CONCLUSIONS: Our results suggest that the flower essence She Oak could have an anxiolytic effect in OVX rats, but that the combination therapy of the She Oak and Bush Fuchsia could avoid the effects of the She Oak.

Effects of gamma-hexachlorocyclohexane and L-3,3',5-triiodothyronine on rat liver cytochrome P4502E1-dependent activity and content in relation to microsomal superoxide radical generation.

Liver microsomal cytochrome P4502E1-dependent p-nitrophenol (PNP) hydroxylation and expression of cytochrome P4502E1 were studied in rats subjected to gamma-hexachlorocyclohexane (HCCH) or L-3,3,5-triiodothyronine (T3) administration as a possible mechanism contributing to superoxide radical (O2.-) generation. HCCH treatment (a single dose of 40 mg/kg body wt) produced a 43% increase in the content of total cytochrome P450, whereas T3 (daily doses of 0.1 mg/kg body wt for two consecutive days) led to a 37% decrease. NADPH-dependent O2.- generation was elevated by HCCH and T3, expressed as either per mg of protein or per nmol of cytochrome P450, with a 135% enhancement in the O2.- production/superoxide dismutase (SOD) activity ratios being observed in both conditions. This was partly due to depression of SOD activity. Concomitantly, the molecular activity of NADPH-cytochrome p450 reductase was enhanced by 90 and 69% by HCCH and T3, respectively. In these conditions, microsomal PNP hydroxylation showed increases of 58 and 45% in HCCH- and T3-treated rats over control values, respectively, with a parallel 31% (HCCH) and 41% (T3) enhancement in the content of cytochrome P4502E1 assessed by western immunoblotting. We conclude that HCCH and T3 enhance the expression and activity of cytochrome P4502E1 and that of NADPH-cytochrome P450 reductase in rat liver, regardless of the changes in total cytochrome P450 content, representing major contributory mechanisms to microsomal NADPH-dependent O2.- generation.