Alpana Sharma

Leptin, IL‐10 and Inflammatory Markers (TNF‐α, IL‐6 and IL‐8) in Pre‐Eclamptic, Normotensive Pregnant and Healthy Non‐Pregnant Women

Despite progress in immunobiology, pre‐eclampsia (PE) remains one of the most common reasons for women to die during pregnancy. The widespread pathophysiological mechanisms are endothelial dysfunction, oxidative stress and inflammation. The aim of this study was to assess the alteration in the levels of leptin, interleukin (IL)‐10 and inflammatory cytokines [tumour necrosis factor (TNF)‐α, IL‐6 & IL‐8] in pre‐eclamptic (severe and mild), healthy pregnant and non‐pregnant women and correlate these parameters with disease severity.

Oxidative stress markers and antioxidant levels in normal pregnancy and pre‐eclampsia

Objective: To compare the levels of 3 oxidative stress markers (glutathione peroxidase [GPX], superoxide dismutase [SOD], and malondialdehyde [MDA]) and 2 antioxidants (vitamin C and lycopene) in healthy and pre‐eclamptic pregnant women.

Biomarkers of Cardiac Injury: An Update:

Conventional and promising new markers of myocardial injury have become an important diagnostic tool and their prognostic significance is also recognized. In addition, they help identify patients who will derive the most benefit from therapeutic interventions. The literature was searched from the websites of the National Library of Medicine (http://www.ncbi.nlm.nih.gov/) and PubMed Central, the U.S. National Library of Medicine’s digital archive of life sciences journal literature (http://www.pubmedcentral.nih.gov/). The data were accessed from books and journals that published relevant articles in this field. The diagnosis of acute myocardial infarction (AMI) has traditionally relied on the combination of chest pain, ECG features, and elevation in serum markers. However, chest symptoms are frequently atypical or absent and ECG changes may be nonspecific or absent. Hence, the diagnosis of acute coronary syndromes has become increasingly dependent on serum markers of cardiac injury. Among them, creatine kinase (CK) is an effective and widely used test, with the recent CKMB assay offering greater specificity and sensitivity. Cardiac troponins facilitate early and rapid diagnosis, enable effective risk stratification in patients with AMI (with or without traditional criteria for MI), and identify those who will benefit from aggressive medical or surgical intervention. Recent data suggest the potential of myoglobin and CKMB isoforms as sensitive markers in the early hours after symptom onset. Cardiac-specific troponins help in rapid diagnosis, prognostication, and treatment of AMI. Troponins also facilitate early detection of recent infarction owing to their prolonged diagnostic window and also aid in the detection of “microinfarction.” CKMB is used to detect reinfarction or infarct extension, if levels rise again after declining. Finally, novel biochemical markers are receiving attention in ongoing trials. They may prove to be more effective in diagnosis and prognosis than their existing counterparts.

Circulatory levels of antioxidants and lipid peroxidation in Indian patients with generalized and localized vitiligo

Vitiligo is an acquired skin disease, characterized by white areas on the skin due to loss of functional melanocytes. The pathogenesis of the disease is still unclear. Published data show the involvement of oxidative stress in the pathophysiology of vitiligo. A total of 30 vitiligo patients and 30 healthy controls were included in this study. We estimated serum levels of malondialdehyde (MDA), vitamins E and C, total antioxidant activity and whole blood levels of superoxide dismutase (SOD) and glutathione peroxidase (GPx) in vitiligo patients and controls. We found significantly higher levels of MDA and significantly lower levels of SOD, GPx, vitamins C and E and total antioxidant activity in vitiligo patients compared with controls. This study is a maiden attempt to report on antioxidant parameters of both generalized/localized-type Indian vitiligo patients. Our results confirmed that oxidative stress may play an important role in the pathogenesis of vitiligo and cause melanocyte damage in vitiligo.

Altered antioxidant status and lipid peroxidation in Indian patients with urothelial bladder carcinoma

OBJECTIVES Urothelial carcinoma of bladder is the second most common urological malignancy after prostate cancer. Recently, there has been increased interest in research of the role of free radicals and antioxidant materials in the prevention, treatment, and alleviation of therapy-related side effects of cancer. In the present study, we aimed to assess the alterations in the levels of antioxidant vitamins, activities of defense enzymes, circulating lipid peroxide, and total antioxidant activity (AOA) in patients with urothelial carcinoma of bladder and correlate these changes with the grade and severity of the disease. MATERIALS AND METHODS The study cohort consisted of 90 subjects; 50 patients with bladder UC (25, low grade; 10, high grade; 15, muscle invasive) and 40 healthy controls. Vitamins C and E, malondialdehyde (MDA), and AOA were estimated using standard protocols. Superoxide dismutase (SOD) and glutathione peroxidase (GPx) were assayed using commercially available kits. RESULTS The serum levels of vitamins C and E, whole blood levels of SOD and GPx, and serum AOA was significantly lower (P < 0.001) while serum MDA levels were significantly higher (P < 0.001) in patients than in controls, indicating presence of oxidative stress in bladder UC patients. The levels of all the biochemical parameters were correlated with the grade and severity of the disease. There were significant differences between the patients with low grade tumors and muscle invasive tumors for all parameters (P < 0.001); except AOA (P < 0.279). CONCLUSIONS The observed redox imbalance in UC of bladder in correlation with the grade and stage, as a consequence of decreased levels of antioxidant vitamins, enzymes, and AOA, along with increased MDA levels in circulation, may be important factors in tumor development and growth. Our results suggest that with advancing stage of bladder UC, the levels of oxidative stress increase, while levels of antioxidant molecules decrease. These findings suggest possible use of antioxidant supplementation as prophylactic agents for prevention and treatment of bladder cancer.

Study of antioxidant levels in patients with multiple myeloma.

Multiple myeloma (MM), neoplastic disorder, is a B-cell malignancy characterised by the accumulation of clonal population of plasma cells. Reactive oxygen species and other free radicals mediate phenotypic and genotypic changes leading from mutation to neoplasia in all cancers including MM. In the present study, 50 clinically diagnosed patients with MM at stage II of international staging system and 50 healthy controls were included. β2 microglobulin levels were estimated by ELISA. The circulating levels of enzymatic antioxidants; superoxide dismutase (SOD), glutathione peroxidase (GPX) were spectrophotometrically estimated using RANDOX kits whereas catalase, malondialdehyde (MDA), vitamin C and E were estimated by standardised protocols using spectrophotometer/fluorometer. The serum β2 microglobulin levels were significantly higher (>3 µg/mL) in patients with MM than healthy controls. The estimated levels of SOD, GPX and catalase (enzymatic antioxidants) and vitamin C and E (non-enzymatic antioxidants) were significantly declined in patients whereas MDA levels were elevated as compared with controls. These results suggest that MM is closely associated with oxidative stress and reduced antioxidant capacity and further investigation might provide an insight to understand a putative causal link between oxidative stress and MM disease progression.

Antioxidant status in advanced cervical cancer patients undergoing neoadjuvant chemoradiation.

Abstract Cervical cancer is the most common cancer in Indian women. The aim of this study is to assess the alterations in the circulating lipid peroxide, antioxidant components and activities of defence enzymes in advanced cervical cancer patients, and to monitor the variations in their levels before and after neoadjuvant chemoradiation. Sixty patients with advanced cancer of the cervix (FIGO IIIa–IVb) are included in the study, along with 60 healthy controls. Blood samples are collected before the start of therapy (S1), two weeks after the second course of chemotherapy (S2) and two weeks after completion of tele/brachyradiation (S3). Single blood samples are taken from controls. Lipid peroxides, conjugated dienes, reduced glutathione (GSH), catalase (CAT) and glutathione-S-transferase (GST) are estimated using standard methods. Glutathione peroxidase (GPx) and superoxide dismutase (SOD) are assayed using commercially available kits. The pretreatment levels of plasma lipid peroxide were significantly elevated in cancer patients, while significantly lowered levels of GSH, GPx, GST, SOD and CAT were observed when compared to controls. After chemotherapy, the levels of lipid peroxidation showed a significant decline (P<0.05), which became highly significant after chemoradiation (P<0.01). Levels of GSH, GPx, SOD, GST and CAT showed a mild increase after chemotherapy. After chemoradiation, levels reverted to normal or near normal (P<0.01). Low levels of antioxidants in the circulation of patients with cervical cancer may be due to their increased utilisation to scavenge lipid peroxidation as well as their sequestration by tumour cells. The observed increase in antioxidant concentration after therapy might be due to the death of tumour cells or the arrest of tumour growth by chemotherapeutic agents. The normalisation of these parameters may provide information about the efficacy of neoadjuvant chemoradiation. A larger patient cohort with a longer follow-up period for therapeutic response studies may yield more significant data.

Involvement of TH1/TH2 Cytokines in the Pathogenesis of Autoimmune Skin Disease—Pemphigus Vulgaris

Pemphigus vulgaris (PV) is classical example of antigen-driven severe autoimmune bullous skin disorder. Auto reactive T cells are critical for the induction and regulation of antibody production. With regard to cytokine production profiles, it has been reported that qualitative as well as quantitative alterations in cytokine production can result in activation of inefficacious effector mechanisms and therefore, complex and severe impairment in immune functions. The purpose of this study was to observe the alterations in the levels of TH1 [Interleukin-2 (IL-2), Interferon-gamma (IFN-γ)] and TH2 (IL-4 and IL-10) cytokines in the sera from patients affected with PV and compared with Pemphigus foliaceus and healthy subjects. This work is aimed to comprehend the involvement of TH1 and TH2 cells as inflammatory infiltrate in the modulation of acantholysis and production of pemphigus lesions. Seventy PV, 13 PF and 50 healthy, age-matched individuals without any generalized skin diseases were included in this study. The diagnosis of PV and PF patients was confirmed by histopathology (hematoxylin and eosin) and / direct immunofluorescence. The levels of TH1 cytokines (IL-2 and IFN-γ) and TH2 cytokine markers (IL-4 and IL-10) were estimated by high sensitivity ELISA kits. All patients with PV and PF showed significantly (p < 0.000) elevated levels of TH2 cytokines (IL-10 and IL-4) as compared with healthy controls. However, the mean concentration of TH1 cytokines (IL-2 and IFN-γ) was significantly decreased in patients as compared to healthy individuals. Both TH1 and TH2 cytokines did not show any significant difference between PV and PF cases. Current concepts support the idea that PV, induced by autoantibodies against Dsg3, is the consequence of an imbalance between Dsg3-reactive TH2 and TH1 cells that may be critical for the maintenance of tolerance against Dsg3. Cytokine profile for confirmed PV cases showed direct evidences for involvement of T cell responses. Increase in IL-4 and IL-10 shows induction of TH2 cells in the pathogenesis of autoimmune disorders Pemphigus vulgaris. The decreased levels of IL-2 and IFN-γ might demonstrate the inhibitory effects by IL-4 and IL-10, which suppress the expansion of TH1 population.

A disproportion of TH1/TH2 cytokines with predominance of TH2, in urothelial carcinoma of bladder ☆

OBJECTIVES Bladder cancer is a common tumor of the urinary tract, accounting for 6% to 8% of all male malignancies and 2% to 3% of all female malignancies. Urothelial carcinoma (UC) of bladder is the second most common urologic malignancy after prostate cancer. Earlier report has elucidated immunologic unreactivity in cancer patients. Cytokines play a pivotal role in the induction of cell mediated and humoral immunity. Quantification of cytokine response in cancer patients can give significant insights about the cellular immunologic potency against the neoplastic cells. In the present study, we aimed to assess alterations of Th1 and Th2 derived cytokines in progression of UC of bladder by determining their circulatory concentration in bladder cancer patients and healthy controls and to correlate the observations with grade and severity of the disease. MATERIALS AND METHODS The study cohort consisted of 122 subjects; 72 patients with bladder UC (28, low grade; 17, high grade; 27, muscle invasive) and 50 healthy controls. The circulatory levels of various cytokines were measured using commercially available sandwich enzyme linked immunosorbent assay (ELISA) kit from BD Biosciences, San Diego, CA, and were statistically correlated according to the grade and the severity of disease. RESULTS The serum levels of typical Th1 cytokines: IL-2 and IFN-γ were found to be significantly lower (P < 0.001) while levels of Th2 cytokines i.e., IL-4, IL-5, and IL-10 were significantly higher (P < 0.001) in patients than in controls. The levels of all the cytokines were correlated with the grade and severity of the disease. There were significant differences between the patients with low grade tumors and muscle invasive tumors for all cytokines (P < 0.001); except IL-10 (P < 0.626). CONCLUSIONS The results of our study delineate that in bladder tumor patients a marked polarization exists towards the expression of Th2 type cytokines while Th1 remain suppressed. Furthermore, the levels of all the cytokines alter according to the grades of the tumor. This can give significant insights about the use of Th1 type cytokines for the administration of immunotherapy to bladder cancer patients. Development of new strategies attempting to manipulate the equilibrium between Th1 and Th2 cells would be beneficial in the management of UC of bladder in future.

Expression of CD147, BIGH3 and Stathmin and their potential role as diagnostic marker in patients with urothelial carcinoma of the bladder

BACKGROUND Urothelial carcinoma of the bladder is characterised by very high recurrence rate, followed up by cystoscopy which being invasive technique makes the need for non-invasive markers important for Transitional Cell Carcinoma (TCC) detection. CD147 is a transmembrane protein highly expressed in tumour cells which aids in tumour invasion and growth. BIGH3, an Extracellular matrix protein (ECM) which interacts with various ECM component in different tissue system and Stathmin(STMN1) is cytosolic microtubule destabilising protein also called as Oncoprotein18 due to its role in tumour promotion. So far the expression of BIGH3 and STMN1 remains undetermined in cancer subjects including TCC. We therefore studied the levels and molecular expression of these molecules in TCC patients, to evaluate their usefulness as diagnostic markers. METHODS Thirty consecutive TCC patients and two sets of control- 15 Benign prostatic hyperplasia (BPH) patient and 15 healthy were taken. Serum and urine levels of these molecules were estimated by ELISA and relative mRNA expression by Q-PCR from tumour and normal urothelium. Post-Hoc analysis and ROC curve were determined to evaluate the significance and sensitivity and specificity. RESULTS The mean concentrations of these molecules were found to be significantly increased (p<0.001) in the serum and urine of TCC patients, with varying significance in each grade for different molecules. The urinary levels of CD147 (67 pg/ml) and serum STMN1 concentration (1.38 ng/ml) showed a specific increase as compared to the controls, while BIGH3 was elevated in both serum and urine samples. Molecular (mRNA) expression was elevated in the high grade (Muscle Invasive) stage of the disease for all the molecules, with a significant 3-fold increase that correlated with disease severity being observed for STMN1. ROC analysis gave optimal combination of sensitivity and specificity for diagnosis of the disease in urine and serum sample for STMN1. CONCLUSION Of CD147, BIGH3 and STMN1, significant results were obtained for STMN1 and it could serve as the best possible diagnostic marker for TCC detection in future.