Biochemical Investigation of Tissue Oxidative Stress and Angiogenesis with Associated Trace Elements in Breast Disease Patients in Uttarakhand, India.

Breast cancer is a heterogeneous cancer with diverse clinical symptoms and an ambiguous molecular spectrum. Oxidative damage, antioxidant activity, and angiogenesis combine to form significant complex factors that stimulate breast cancer development and progression. This study is designed to determine level changes in total antioxidant status and markers of lipid peroxidation melondialdehyde (MDA) and angiogenesis vascular endothelial growth factor (VEGF) along with related micronutrients of copper, zinc, magnesium, and iron in malignant and benign breast disease tissue extracts. We assess specificity and sensitivity of those markers using the area under the curve of receiver operator characteristic (ROC) curve analysis. Association studies are done with correlation analysis. The tissue extract level of MDA markers is found to be significantly higher (14.118 ± 1.47 nmol/g tissue; p < 0.05), with significantly depleted levels of antioxidants (5.983 ± 1.661 nmol/g tissue; p < 0.05). The tissue VEGF level also significantly increases in a diseased condition (512.466 ± 5.661 pg/mg tissue) versus the nondiseased condition (422.433 ± 13.615 pg/mg tissue). Related trace-element levels show a significant mixed pattern among studied groups. VEGF emerges as the best discriminatory biomarker of breast cancer presence, in accordance with ROC analysis. Oxidative stress and angiogenesis are found to be important factors in breast cancer development. This study forms the basis for future studies that focus on the relationship between roles of indices studied and cancer induction.

A novel approach to study oxidative stress in neonatal respiratory distress syndrome.

BACKGROUND: Respiratory distress syndrome of the neonate (neonatal RDS) is still an important problem in treatment of preterm infants. It is accompanied by inflammatory processes with free radical generation and oxidative stress. The aim of study was to determine the role of oxidative stress in the development of neonatal RDS. METHODS: Markers of oxidative stress and antioxidant activity in umbilical cord blood were studied in infants with neonatal respiratory distress syndrome with reference to healthy newborns. RESULTS: Status of markers of oxidative stress (malondialdehyde, protein carbonyl and 8-hydroxy-2-deoxy guanosine) showed a significant increase with depleted levels of total antioxidant capacity in neonatal RDS when compared to healthy newborns. CONCLUSION: The study provides convincing evidence of oxidative damage and diminished antioxidant defenses in newborns with RDS. Neonatal RDS is characterized by damage of lipid, protein and DNA, which indicates the augmentation of oxidative stress. GENERAL SIGNIFICANCE: The identification of the potential biomarker of oxidative stress consists of a promising strategy to study the pathophysiology of neonatal RDS.

Correlation of serum toll like receptor 9 and trace elements with lipid peroxidation in the patients of breast diseases.

Toll-like receptors are recognized as redox sensitive receptor proteins and have been implicated in cellular response to oxidative stress. Altered pro-oxidant-antioxidant balance leads to an increased oxidative damage and consequently play an important role in breast diseases. The study was designed to access the oxidative stress status by quantification of byproducts generated during lipid peroxidation and inadequate trace elements during oxidative damage and its effects on the toll like receptor (TLR) activity in patients of breast diseases. Decreased levels of selenium, copper, zinc, magnesium and iron with elevated levels of malondialdehyde (marker of lipid peroxidation) were accompanied by decreased TLR activity in patients of benign breast diseases as well as breast carcinoma. A similar pattern was observed with the advancement of disease and its subsequent progression in breast carcinoma patients. Results of multinomial regression analysis suggest benign breast disease patients are at higher risk of developing breast cancer with high odds ratio of lipid damage.

Association between biomarkers of oxidative stress, trace elements, and cell proliferation index in patients with benign and malignant breast diseases.

OBJECTIVES: To understand the association between biomarkers of oxidative stress, antioxidants, trace elements, and cell proliferation index in relation to the disease progression in the pathophysiology of breast diseases. DESIGN AND METHODS: Concentrations of markers of oxidative stress, antioxidants, trace elements, and cell proliferation index were evaluated in the patients with benign breast diseases, malignant breast diseases, and healthy volunteers as controls. Multinomial logistic regression analysis was used to identify the contribution of the selected indexes using odds ratio and associated confidence interval. RESULTS: The level of markers of oxidative stress (malondialdehyde [MDA]) and cell proliferation index were found to be significantly higher with significantly depleted levels of antioxidants and trace elements in breast cancer patients compared with control subjects as well as benign breast disease patients. A similar pattern of changes were observed between benign and control subjects. CONCLUSION: An inadequate amount of antioxidant enzymes and trace elements may be an important contributing factor associated with oxidative stress leading to elevated levels of MDA and cell proliferation index in relation to disease progression and clinical stage in the pathophysiology of breast diseases.

Expression of serum toll-like receptor 9 and oxidative damage markers in benign and malignant breast diseases.

The intracellular redox environment plays an important role in the maintenance of proper cellular homeostasis and functions. Disturbances in redox equilibrium of cells result in pro-inflammatory conditions, and these inflammatory conditions can induce carcinogenesis or increase the malignant potential of the tumor. Oxidative stress or tissue damage can trigger toll-like receptor (TLR) family of receptors that are involved in altering the innate immune system. The present study was aimed at evaluating the level of oxidative damage markers in breast diseases by measuring the 8-hydroxydeoxyguanosine (8-OHdG), protein carbonyl (PC), malondialdehyde (MDA), and total antioxidant status (TAS) alterations in relation to expression of TLR-9. A significant increase in the level of oxidative damage markers was observed in breast carcinoma patients in comparison to benign and normal controls, which was accompanied by a significant decrease in TAS and expression of TLR-9 concentrations. 8-OHdG, PC, and MDA were negatively correlated with expression of TLR-9 and TAS levels. Altered levels of biomarkers of oxidative stress and TLR-9 among the malignant, benign, and controls suggest a correlation of oxidative stress and TLR signaling in the progression of disease in breast carcinoma patients. Receiver operating characteristic analysis depicts that expression of TLR-9 is a good indicator for distinguishing cancer patients from benign and normal controls. High accuracy, specificity, and sensitivity of oxidative stress markers and expression of TLR-9 can be used as discriminatory marker/s for efficient diagnosis.

Association of oxidative DNA damage, protein oxidation and antioxidant function with oxidative stress induced cellular injury in pre-eclamptic/eclamptic mothers during fetal circulation.

Pre-eclampsia is a devastating multi system syndrome and a major cause of maternal, fetal, neonatal morbidity and mortality. Pre-eclampsia is associated with oxidative stress in the maternal circulation. To have an insight on the effect of pre-eclampsia/eclampsia on the neonates, the study was made to explore the oxidative status by quantification of byproducts generated during protein oxidation and oxidative DNA damage and deficient antioxidant activity in umbilical cord blood of pre-eclamptic/eclamptic mothers during fetal circulation. Umbilical cord blood during delivery from neonates born to 19 pre-eclamptic mothers, 14 eclamptic mothers and 18 normotensive mothers (uncomplicated pregnancy) as control cases was collected. 8-OHdG (8-hydroxy-2-deoxyguanosine), protein carbonyl, nitrite, catalase, non-enzymatic antioxidants (vitamin A, E, C), total antioxidant status and iron status were determined. Significant elevation in the levels of 8-OHdG, protein carbonyl, nitrite and iron along with decreased levels of catalase, vitamin A, E, C, total antioxidant status were observed in the umbilical cord blood of pre-eclamptic and eclamptic pregnancies. These parameters might be influential variables for the risk of free radical damage in infants born to pre-eclamptic/eclamptic pregnancies. Increased oxidative stress causes oxidation of DNA and protein which alters antioxidant function. Excess iron level and decreased unsaturated iron binding capacity may be the important factor associated with oxidative stress and contribute in the pathogenesis of pre-eclampsia/eclampsia which is reflected in fetal circulation.

Simultaneous progression of oxidative stress, angiogenesis, and cell proliferation in prostate carcinoma.

OBJECTIVES: To understand the association between markers of oxidative stress, levels of vascular endothelial growth factor (VEGF), and cell proliferation index in relation to disease progression, clinical stage, and cytologic grade in pathophysiology of prostate carcinoma. PATIENTS AND METHODS: Case control study comprised of 40 prostate carcinoma patients along with 40 age- and sex-matched healthy subjects as controls. Levels of 8-hydroxy-2-deoxy guanosine, protein carbonyl, and malondialdehyde along with total antioxidant status were measured to study the oxidative stress status in the study subjects. Angiogenesis was evaluated by studying the VEGF level and cell proliferation index. RESULTS: The levels of markers of oxidative stress along with VEGF and cell proliferation index were found to be significantly higher with significantly decreased levels of antioxidant activity in the study subjects in comparison with healthy controls. The results indicate oxidative stress, angiogenesis, and cell proliferation activity increase progressively with the increase in staging and progression of disease. CONCLUSIONS: Oxidative stress parameters, angiogenesis, and cell proliferation activity point clearly that with the progression of oxidative stress there is a simultaneous progression of angiogenesis, regulation and control of endothelial cell proliferation in relation to disease progression, clinical stage, and cytologic grade in the pathophysiology of prostate carcinoma.

Trace elements and antioxidant enzymes associated with oxidative stress in the pre-eclamptic/eclamptic mothers during fetal circulation.

BACKGROUND & AIMS: Pre-eclampsia is associated with oxidative stress in the maternal circulation. The aim of the study was to access the oxidative stress status by quantification of byproducts generated during lipid peroxidation; deficient antioxidant activity and inadequate trace elements during oxidative damage in the umbilical cord blood of pre-eclamptic/eclamptic mothers and its effect on the fetal outcome. METHODS: In a case control study umbilical cord blood samples were collected during delivery from cases of pre-eclamptic/eclamptic mothers and from normotensive (uncomplicated pregnancy) subjects as controls. Concentrations of malondialdehyde, trace elements (Zn, Cu, Se, Mg) and antioxidant enzymes (SOD, GPx) were determined. RESULTS: Decreased levels of Zinc (p < 0.001), Copper (p < 0.001), Selenium (p < 0.005), Magnesium (p < 0.05), Superoxide dismutase (p < 0.001), Glutathione Peroxidase (p < 0.001) and elevated levels of malondialdehyde (marker of lipid peroxidation) in the umbilical cord blood of pre-eclamptic and eclamptic pregnancies were observed. Positive correlation between Zn and SOD (Pearson r = 0.581, p < 0.001), Cu and SOD (Pearson r = 0.576, p < 0.001) and Se and GPx (Pearson r = 0.445, p < 0.005) was also observed. CONCLUSIONS: The reduced levels of trace elements associated with inadequate amount of antioxidant enzymes may be important contributing factor associated with oxidative stress leading to endothelial dysfunction in pre-eclamptic/eclamptic mothers.

In vivo oxidative DNA damage and lipid peroxidation as a biomarker of oxidative stress in preterm low-birthweight infants.

OBJECTIVES: To analyze the status of oxidative stress in relation to the degree of prematurity and birthweight of neonates. MATERIALS AND METHODS: Umbilical cord blood samples were obtained at the time of delivery. 8-Hydroxy-2-deoxy guanosine (8-OHdG) has been measured as oxidative DNA damage marker in preterm low-birthweight (LBW) newborns by competitive in vitro enzyme-linked immunosorbent assay (ELISA) along with malondialdehyde (MDA) as marker of lipid peroxidation and total antioxidant status to study the oxidative stress. RESULTS: Significant elevation in the levels of 8-OHdG along with malondialdehyde has been noted in preterm LBW newborns. Serum 8-OHdG is found to be significantly and negatively correlated with birthweight (r = -0.834, p < 0.001) and gestational age of the newborn (r = -0.626, p < 0.001). CONCLUSION: These results provide evidence of increased oxidative stress in the form of DNA damage and lipid peroxidation in premature LBW newborns, which may be responsible for different complications associated with prematurity.

Vascular endothelial growth factor levels in relation to oxidative damage and antioxidant status in patients with breast cancer.

PURPOSE: Oxidative stress and angiogenesis are important elements in the pathogenesis of inflammatory diseases and cancer. Vascular endothelial growth factor (VEGF) is one of the most potent angiogenic cytokines and is up-regulated by conditions associated with the generation of free radicals and reactive oxygen intermediates. In this study, we investigated the association between oxidative stress and serum VEGF status in patients with breast cancer. METHODS: Forty patients with breast carcinoma, of which 21 were stage II and 19 were stage III, along with 40 age- and gender-matched healthy controls were enrolled. Oxidative stress, total antioxidant status, and VEGF levels in serum were evaluated by spectrophotometric procedures. Malondialdehyde (MDA) levels were measured and antioxidant status was assessed by measuring total antioxidant status (TAS) to assess oxidative damage. RESULTS: VEGF and MDA levels were significantly higher in patients with breast cancer than those of controls (p<0.005). Total antioxidant level decreased significantly in patients compared to that in controls. MDA, TAS, and VEGF levels were also analyzed based on menopausal status and different clinical disease stages. MDA and TAS level significantly different in the postmenopausal group than the premenopausal group, whereas VEGF level remained unchanged. CONCLUSION: Increased VEGF level and its positive correlation with oxidative stress level and decreased antioxidant status suggest a link between oxidative stress and malignant transformation.

Evaluation of oxidative stress status following polyherbal formulation therapy in patients of cholelithiasis with choledocholithiasis.

Free radicals produce persistent oxidative stress in biological system and are highly reactive molecules produced as a byproduct of metabolism. A reactive free redical generated in the body reacts with non-radical molecules and results in free radical chain reaction leading to formation of new free radicals. If the defense mechanism of body fails to combat them or they are not properly utilized in the body -these silent killers pose a threat by injuring tissues, their proteins and fat contents. Lipids in the cell membrane undergo degradation to form hydroperoxides((1, 2, 3).) Polyunsaturated fatty acids, PUFA, are especially liable to lipid peroxidation. Lipid hydroperoxids decompose to form a variety of products including malondialdehyde (MDA) which is used as an indicator of oxidative damage of cells and tissues((4).)The present investigations involve the study of oxidative stress in the bile juice from the patients of cholecystitis/cholelithiasis with choledocholithiasis treated by cholecystectomy with choledochotomy (CBD exploration) with T-tube drainage. Results of malondialdehyde status in the bile juice of these patients pre-operatively and following polyherbal formulation therapy from 3(rd) to 10(th) post operative day are discussed.