Abhigyan Satyam

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.

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.

Macromolecular Crowding Meets Tissue Engineering by Self‐Assembly: A Paradigm Shift in Regenerative Medicine

MMC, the addition of inert polydispersed macromolecules in the culture media, effectively emulates the dense in vivo extracellular space, resulting in amplified deposition of ECM in vitro and subsequent production of cohesive, ECM-rich living substitutes.

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.

Engineering in vitro microenvironments for cell based therapies and drug discovery

Traditional ex vivo culture setups fail to imitate the native tissue niche, leading to cellular senescence, phenotypic drift, growth arrest and loss of stem cell multipotency. Growing evidence suggests that surface topography, substrate stiffness, mechanical stimulation, oxygen tension and localised density influence cellular functions and longevity, enhance tissue-specific extracellular matrix deposition and direct stem cell differentiation. In this review, we discuss how these cues will facilitate engineering of physiological in vitro microenvironments to enable clinical translation of cell based therapies and development of in vitro models for drug discovery applications.

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.

Macromolecularly crowded in vitro microenvironments accelerate the production of extracellular matrix-rich supramolecular assemblies

Therapeutic strategies based on the principles of tissue engineering by self-assembly put forward the notion that functional regeneration can be achieved by utilising the inherent capacity of cells to create highly sophisticated supramolecular assemblies. However, in dilute ex vivo microenvironments, prolonged culture time is required to develop an extracellular matrix-rich implantable device. Herein, we assessed the influence of macromolecular crowding, a biophysical phenomenon that regulates intra- and extra-cellular activities in multicellular organisms, in human corneal fibroblast culture. In the presence of macromolecules, abundant extracellular matrix deposition was evidenced as fast as 48 h in culture, even at low serum concentration. Temperature responsive copolymers allowed the detachment of dense and cohesive supramolecularly assembled living substitutes within 6 days in culture. Morphological, histological, gene and protein analysis assays demonstrated maintenance of tissue-specific function. Macromolecular crowding opens new avenues for a more rational design in engineering of clinically relevant tissue modules in vitro.

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.