NOPR Collection: [Pages 667-744]

Compositional alterations in erythrocyte membranes in Type II diabetes

2020-09-30T09:16:16Z

Title: Compositional alterations in erythrocyte membranes in Type II diabetes Authors: Bhise, Sunita S.; Rao, Janhavi R.; Hegde, Mahabaleshwar V.; Katyare, Surendra S. Abstract: Loss of erythrocyte membrane deformability is one of the most crucial factors in developing complications associated with Type II diabetes. The observed loss of erythrocyte membrane deformability could be related to structural changes in the membrane. In this context, here, we have made an attempt at gaining a better insight (quantitative as well as qualitative) into the protein and lipid contents in erythrocyte membranes and their interrelationships in Type II diabetes. Age matched control (n=12) and Type II diabetic subjects (n=22) were selected for this study. Morphological characteristics were studied by atomic force microscopy (AFM). AFM study confirmed remarkable alterations in morphology of the diabetic erythrocytes. In diabetic erythrocytes following changes were noted: (i) Significant increase in membrane as well as cytosolic proteins with a marginal increase in phospholipids content; (ii) The membrane total lipids:protein, phospholipids:protein, cholesterol:protein and phospholipids:cholesterol (mole:mole) ratios decreased significantly; (iii). A reproducible decrease in docosahexaenoic acid (DHA) and Omega-3 index with increase in Omega-6:Omega-3 ratio in membrane fatty acids; and (iv) The SDS-PAGE analysis indicated that all membrane proteins increased in almost equal proportion leading to increased membrane protein content. The observed compositional and stochiometric changes in lipids, proteins and their ratios may underlie morphological alterations and loss of deformability. Page(s): 671-679

Low dose of 131I-F(ab')2-Rituximab and 131I-Rituximab induces G1arrest and apoptosis in Raji cells (Burkitt’s lymphoma)

2020-09-30T09:13:13Z

Title: Low dose of 131I-F(ab')2-Rituximab and 131I-Rituximab induces G1arrest and apoptosis in Raji cells (Burkitt’s lymphoma) Authors: Suman, Shishu Kant; Kameswaran, Mythili; Dash, Ashutosh Abstract: Radiolabeled fragmented F(ab')₂ antibodies had shown better therapeutic efficacy than radiolabeled intact antibodies in treating cancers. In this study, we investigated the differences and similarities on the mechanism and extent of cell death in Raji cells (Burkitt’s lymphoma) in response to 370 kBq of ¹³¹I-F(ab')₂-Rituximab and ¹³¹I-Rituximab up to 72 h. F(ab')₂ of Rituximab was prepared and characterized by SE-HPLC and SDS-PAGE. Fragmented and intact Rituximab were radioiodinated by Chloramine-T method. Toxicity and mechanism of cell death in Raji cells in response to ¹³¹I-F(ab')₂-Rituximab and ¹³¹I-Rituximab were studied by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), LDH (lactate dehydrogenase), trypan blue exclusion, viability, apoptotic, caspase assays and cell cycle analysis. The cytotoxicity assays showed slow death of Raji cells up to 24 h in response to both ¹³¹I-F(ab')₂-Rituximab and ¹³¹I-Rituximab. Cell cycle analysis at 30 h showed G1 arrest in Raji cells which led to its slow cell death up to 24 h. Elucidative assays to identify the molecular mechanism of death of G1arrested Raji cells showed apoptotic cell death at 40 h after treatment, which was validated by demonstrating caspase activation in arrested Raji cells. Toxicity studies and mechanism of cell death in Raji cells demonstrated comparable results when treated with equivalent doses (370 kBq) of radiolabeled antibodies indicating ¹³¹I-F(ab')₂-Rituximab as a potential radioimmunotherapeutic agent for patients with Non-Hodgkin’s lymphoma. Page(s): 680-690

Amelioration of cisplatin induced nephrotoxicity by Phyla nodiflora (L.) Greene

2020-09-30T09:10:32Z

Title: Amelioration of cisplatin induced nephrotoxicity by Phyla nodiflora (L.) Greene Authors: Sharma, Sonam; Dixit, Priya; Sairam, K; Sahu, Alakh N Abstract: Phyla nodiflora (Verbenaceae), commonly called frog fruit and locally, jal bhuti, is the traditional folk medicine accepted in the Indian Medicine as well as the Traditional system of Chinese Medicine (TCM) for various treatments viz., urinary disorder, lithiasis, knee joint pain, diuresis and swelling. In the present study, we tried to standardise crude Phyla nodiflora ethanol extract (PNE) using HPTLC and also evaluated its protection against cisplatin induced nephrotoxicity in rats. HPTLC quantification of rutin and fingerprinting profile was performed. Serum kidney toxicity markers, renal tissue antioxidant and pro-inflammatory cytokine levels were assessed followed by DNA fragmentation assay and histopathological examination of renal tissue. Rutin concentration in ethanol extract was found to be 5.35% w/w. In HPTLC fingerprinting 12 peaks with R_f ranges from 0.08 to 0.86 were confirmed. Serum biochemical parameters, renal tissue antioxidants and pro-inflammatory cytokines levels were found to be restored. The results demonstrated the nephroprotective activity of P. nodiflora ethanol extract (400 mg/kg, p.o.) as evidenced by protection of kidneys from cisplatin induced DNA fragmentation and damage tissue architecture. Page(s): 691-698

Antileishmanial apoptotic activity of Nigella sativa L. essential oil and thymoquinone triggers on Leishmania tropica

2020-09-30T09:08:12Z

Title: Antileishmanial apoptotic activity of Nigella sativa L. essential oil and thymoquinone triggers on Leishmania tropica Authors: Al-Turkmani, Mohamad Osama; Mokrani, Loubna; Soukkarieh, Chadi Abstract: Nigella sativa L., commonly called Black cumin, is well-known in folk medicine and numerous studies have shown its various pharmacological activities. In this study, we estimated the cytotoxic effects of N. sativa essential oil (NEO) and its major bioactive component Thymoquinone (TQ) on Leishmania tropica promastigotes that cause cutaneous leishmanias, and also observed the programmed cell death features. The extraction of NEO was done by hydro-distillation and analyzed by Gas Chromatography-Mass Spectrometry (GC-MS). Antileishmanial activity of NEO and TQ was determined using 3-(4.5-dimethylthiazol-2-yl)-2.5-diphenyl tetrazolium bromide (MTT) assay and the obtained results are expressed as 50% inhibitory concentration (IC₅₀). The leishmanicidal activity of NEO and TQ was mediated via apoptosis as evidenced by in situ labelling of DNA fragments using terminal deoxyribonucleotidyltransferase-mediated dUTP nick end labelling (TUNEL) and cell cycle arrest at sub G₀/G₁ phase. The IC₅₀ values were 5 µg/mL and 1.3 µg/mL, respectively. We noted a significant increase in DNA fragmentation in treated parasites with IC₅₀ of both NEO and TQ as well as a cell cycle arrest. These results revealed that NEO and TQ possess potential antileishmanial activity that mediated high possibility by programmed cell death. Page(s): 699-705