Immunomodulatory Role of Ocimum gratissimum and Ascorbic Acid against Nicotine-Induced Murine Peritoneal Macrophages In Vitro

The aim of this present study was to evaluate the immune functions and immune responses in nicotine-induced (10 mM) macrophages and concurrently establish the immunomodulatory role of aqueous extract of Ocimum gratissimum (Ae-Og) and ascorbic acid. In this study, nitrite generations and some phenotype functions by macrophages were studied. Beside that, release of Th1 cytokines (TNF-α, IL-12) and Th2 cytokines (IL-10, TGF-β) was measured by ELISA, and the expression of these cytokines at mRNA level was analyzed by real-time PCR. Ae-Og, at a dose of 10 μg/mL, significantly reduced the nicotine-induced NO generation and iNOSII expression. Similar kinds of response were observed with supplementation of ascorbic acid (0.01 mM). The administration of Ae-Og and ascorbic acid increased the decreased adherence, chemotaxis, phagocytosis, and intracellular killing of bacteria in nicotine-treated macrophages. Ae-Og and ascorbic acid were found to protect the murine peritoneal macrophages through downregulation of Th1 cytokines in nicotine-treated macrophages with concurrent activation of Th2 responses. These findings strongly enhanced our understanding of the molecular mechanism leading to nicotine-induced suppression of immune functions and provide additional rationale for application of anti-inflammatory therapeutic approaches by O. gratissimum and ascorbic acid for different inflammatory disease prevention and treatment during nicotine toxicity

Age associated oxidative damage in lymphocytes

Lymphocytes are an important immunological cell and have been played a significant role in acquired immune system; hence, may play in pivotal role in immunosenescence. Oxidative stress has been reported to increase in elderly subjects, possibly arising from an uncontrolled production of free radicals with aging and decreased antioxidant defenses. This study was aimed to evaluate the level of lipid-protein damage and antioxidant status in lymphocytes of healthy individuals to correlate between oxidative damage with the aging process. Twenty healthy individuals of each age group (11–20; 21–30; 31–40; 41–50; and 51–60 years) were selected randomly. Blood samples were drawn by medical practitioner and lymphocytes were isolated from blood samples. Malondialdehyde (MDA), protein carbonyls (PC) level were evaluated to determine the lipid and protein damage in lymphocytes. Superoxide dismutase (SOD), catalase (CAT), glutathione and glutathione dependent enzymes were estimated to evaluate the antioxidant status in the lymphocytes. Increased MDA and PC levels strongly support the increased oxidative damage in elderly subject than young subjects. The results indicated that, balance of oxidant and antioxidant systems in lymphocytes shifts in favor of accelerated oxidative damage during aging. Thus oxidative stress in lymphocytes may particular interest in aging and may play important role in immunosenescence

Internalization of Staphylococcus aureus in Lymphocytes Induces Oxidative Stress and DNA Fragmentation: Possible Ameliorative Role of Nanoconjugated Vancomycin

Staphylococcus aureus is the most frequently isolated pathogen causing bloodstream infections, skin and soft tissue infections and pneumonia. Lymphocyte is an important immune cell. The aim of the present paper was to test the ameliorative role of nanoconjugated vancomycin against Vancomycin-sensitive Staphylococcus aureus (VSSA) and vancomycin-resistant Staphylococcus aureus (VRSA) infection-induced oxidative stress in lymphocytes. VSSA and VRSA infections were developed in Swiss mice by intraperitoneal injection of 5 × 106 CFU/mL bacterial solutions. Nanoconjugated vancomycin was adminstrated to VSSA- and VRSA-infected mice at its effective dose for 10 days. Vancomycin was adminstrated to VSSA- and VRSA-infected mice at a similar dose, respectively, for 10 days. Vancomycin and nanoconjugated vancomycin were adminstrated to normal mice at their effective doses for 10 days. The result of this study reveals that in vivo VSSA and VRSA infection significantly increases the level of lipid peroxidation, protein oxidation, oxidized glutathione level, nitrite generation, nitrite release, and DNA damage and decreases the level of reduced glutathione, antioxidant enzyme status, and glutathione-dependent enzymes as compared to control group, which were increased or decreased significantly near to normal in nanoconjugated vancomycin-treated group. These findings suggest the potential use and beneficial role of nanoconjugated vancomycin against VSSA and VRSA infection-induced oxidative stress in lymphocytes

Attenuation of Mycobacterium species through direct and macrophage mediated pathway by unsymmetrical diaryl urea

Tuberculosis is a major threat for mankind and the emergence of resistance strain of Mycobacterium tuberculosis (Mtb) against first line antibiotics makes it lethal for human civilization. In this study, we have synthesized different diaryl urea derivatives targeting the inhibition of mycolic acid biosynthesis. Among the 39 synthesized molecules, compounds 46, 57, 58 and 86 showed MIC values ≤ 10 μg/ml against H37Rv and mc26030 strains. The best molecule with a methyl at ortho position of the first aromatic ring and prenyl group at the meta position of the second aromatic ring showed the MIC value of 5.2 μg/ml and 1 μg/ml against H37Rv and mc26030 respectively, with mammalian cytotoxicity of 163.4 μg/ml. The effective compounds showed selective inhibitory effect on mycolic acid (epoxy mycolate) biosynthesis in14C-radiolabelled assay. At the same time these molecules also executed their potent immunomodulatory activity by up-regulation of IFN-γ and IL-12 and down-regulation of IL-10.Fil: Velappan, Anand Babu. Sastra University; IndiaFil: Charan Raja, Mamilla R.. Sastra University; IndiaFil: Datta, Dhrubajyoti. Indian Institute of Science Education and Research Pune; IndiaFil: Tsai, Yi Ting. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Halloum, Iman. Université de Montpellier; Francia. Centre National de la Recherche Scientifique; FranciaFil: Wan, Baojie. University of Illinois; Estados UnidosFil: Kremer, Laurent. Université de Montpellier; Francia. Inserm; Francia. Centre National de la Recherche Scientifique; FranciaFil: Gramajo, Hugo Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Franzblau, Scott G.. University of Illinois; Estados UnidosFil: Kar Mahapatra, Santanu. Sastra University; IndiaFil: Debnath, Joy. Sastra University; Indi

Smoking induced oxidative stress in serum and neutrophil of the university students

Cigarette smoking has been implicated as a significant risk factor for the establishment and progression of several diseases. The purpose of this study was to examine the effect of smoking in serum as well as in neutrophil oxidative damage and antioxidant status. Subjects were randomly chosen from male university students (22-25 years) in a rural community in the District of Midnapore, West Bengal, India. To understand status of oxidative damage, we measured the level of lipid peroxidation (MDA), activities of lactate dehydrogenase (LDH), catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), and reduced glutathione (GSH) level. The MDA level and the LDH activity in serum and neutrophil were higher in smokers than the non-smokers. The activities of CAT, SOD, GPx, GR and the level of GSH were significantly lower in smokers in comparison to the non-smokers in both serum and neutrophil. These findings may suggest that, students with a short smoking history have evidence of oxidative stress and an impaired oxidant defense system. Alterations observed in smokers that increased oxidative stress can represent a risk factor for the development of chronic disease in earlier future

In Vitro Time Dependent Nicotine-Induced Free Radical Generation and Status of Glutathione Cycle in Murine Peritoneal Macrophage

: Nicotine is an alkaloid and the precursor to many tobacco carcinogens present in various tobacco products. The aim of the present study was to evaluate the free radical generation and status of the glutathione cycle in murine peritoneal macrophage during in vitro nicotine exposure with different time interval. Peritoneal macrophages were treated with 10mM nicotine in vitro for different time interval (3, 6, 9, 12, 15, 18, & 24h) and super oxide anion generation and components of glutathione cycle were analyzed. Super oxide anion generation, and NADPH oxidase activity got peak at 12hr, indicates maximum free radical generation through activation of NADPH oxidase in murine peritoneal macrophages during nicotine treatment. Reduced glutathione level, glutathione peroxidase, glutathione reductase, and glutathione-S-transferase activity were decreased significantly (p<0.05) with increasing time of nicotine treatment. But the oxidized glutathione level was time dependently increased significantly (p<0.05) in murine peritoneal macrophages. The redox ratio also decreased significantly (p<0.05) at all time of nicotine treatment. All the changes in peritoneal macrophages after 12hr in vitro nicotine treatment had no significant difference. From this study, it may be summarized that nicotine not only generates excess free radical but also affect the glutathione cycle in murine peritoneal macrophage

Aqueous extract of Ocimum gratissimum Linn and ascorbic acid ameliorate nicotine–induced cellular damage in murine peritoneal macrophage

AbstractObjectiveTo test the in vitro protective role of aqueous extract of Ocimum gratissimum Linn. (O. gratissimum) and ascorbic acid against nicotine-induced murine peritoneal macrophage.MethodsPeritoneal macrophages from mice were treated with nicotine (10 mM), nicotine (10 mM) with aqueous extract of O. gratissimum (1 to 25 μg/mL), and nicotine (10 mM) with ascorbic acid (0.01 mM) for 12 h in cell culture media, while the control group was treated with culture media. Levels of free radical generation, lipid peroxidation, protein carbonyls, oxidized glutathione levels and DNA damage were observed and compared.ResultsPhytochemical analysis of aqueous extract has shown high amount of phenolics and flavonoids compound present in it. The significantly increased free radical generation, lipid peroxidation, protein carbonyls, oxidized glutathione levels and DNA damage were observed in nicotine-treated group as compared to the control group; those were significantly reduced in aqueous extract of O. gratissimum and ascorbic acid supplemented groups. Moreover, significantly reduced antioxidant status in nicotine exposed murine peritoneal macrophage was effectively ameliorated by these two products. Among the different concentration of aqueous extract of O. gratissimum, the maximum protective effect was observed at 10 μg/mL which does not produce any significant change in the normal cell.ConclusionsThese findings suggest the potential use and beneficial role of O. gratissimum as a modulator of nicotine-induced cellular damage in murine peritoneal macrophage

VERSATILE AND SYNERGISTIC POTENTIAL OF EUGENOL

Eugenol (1-allyl-4-hydroxy-3-methoxybenzene) is the phenolic component of essential oil and the main constituent of Eugenia caryophyllata, Ocimmum gratissimum and several others medicinal plant. In view of its non-mutagenic and non-carcinogenic properties, eugenol is generally regarded as safe by the Food and Agricultural Organization of the United Nations. Eugenol has been recently shown to be effective for antimicrobials and treatment of different life threatening diseases including sepsis, leishmaniasis, and cancer. However, overall, activity of eugenol is not discussed elsewhere. In this review, we discuss the current understanding of the mechanisms involved the antioxidant, antimicrobial, anticancer and anti-inflammatory potential of eugenol.Â

Progressive increase in point mutations associates chloroquine resistance: Even after withdrawal of chloroquine use in India

Chloroquine (CQ) is highly effective against P. vivax, due to the rapid spread of CQ resistance in P. falciparum parasites; it is no longer the drug of choice against P. falciparum. This study elucidates the scenario of chloroquine efficacy at times that coincided with a new drug policy and especially assessed the chloroquine resistant molecular markers after withdrawal of chloroquine in Kolkata and Purulia, two malaria endemic zones of West Bengal, India. In vitro CQ susceptibility was tested in 781 patients with P. falciparum mono infections between 2008 and 2013, of which 338 patients had received CQ in 2008–2009. Genotyping of the pfcrt and the pfmdr1 gene was carried out in all isolates. Early treatment failure was detected in 114 patients {43 (31·39%) from Kolkata and 71 (35·32%) from Purulia} while recrudescence was identified in 13 (9.49%) and 17 (8.46%) patients from Kolkata and Purulia respectively. In vivo chloroquine resistance was strongly associated with CVMNT-YYSNY (p < 0.01) and SVMNT-YYSNY (p < 0.05) allele in Kolkata. In Purulia chloroquine resistance was associated with CVMNK-YYSNY (P < 0.005), SVMNT-YYSNY (P < 0.01) allele. The proportion of in vitro chloroquine resistance increased in subsequent years to 87.23% and 93·10% in 2013, in Kolkata and Purulia, respectively. Isolates with SVMNT-YFSND, SVMNT-YFSNY, CVIET-YFSND and CVIET-YYSNY haplotypes increased gradually (p < 0.05) from 2010 to 2013, leading to a rise in IC50 (p < 0.05) of chloroquine. An increase in in vitro chloroquine resistance and candidate gene mutations even after five years of chloroquine withdrawal against P. falciparum calls for synchronized research surveillance and proper containment strategies. Keywords: Plasmodium falciparum, ChloroQuine resistance in India, pfcrt polymorphism, pfmdr1 mutation, In vitro chloroquine resistanc