ANTI-INFLAMMATORY ACTIVITY OF AQUEOUS EXTRACTS OF GRACILARIA

Objective: In the present investigation an attempt was made to examine the anti-inflammatory potential of aqueous extracts of Gracilaria salicornia C. Ag., Gracilaria edulis (Gmelin) Silva, Gracilaria corticata J. Ag., Gracilaria fergusonii J. Ag.,and Gracilaria verrucosa (Hudson) Papenfus from Mandapam, Gracilaria edulis (Gmelin) Silva., Gracilaria verrucosa (Hudson) Papenfus from Pulicate Lake, Gracilaria fergusonii J. Ag., Gracilaria corticata J. Ag. and Gracilaria corticata J. Ag. var. cylindrica from Tuticorin using heat induced haemolysis of RBC. In addition, the present study is focused to reveal the locality specific anti-inflammatory activities of selected Gracillaria species.Methods: The fresh seaweeds were boiled with distilled water (1:20 W/V) for 2 h. The slurry was filtered through Whatman No. 41 filter paper and condensed. The semi-solid crude extracts were used for anti-inflammatory analysis.Results: An aqueous extracts of studied Gracilaria species were effective in inhibiting the heat induced haemolysis at different concentrations. The results showed the dose dependent protection. The percentage of anti-inflammatory activity of studied Gracillaria was varied from 43.81 to 95.55. The highest percentage (95.55%) of activity was observed in 250 µg/ml of G. edulis aqueous extracts. The anti-inflammatory activity of studied Gracillaria species at 250 µg/ml were as follows G. edulis (Mandabam)>G. corticata (Mandabam)>G. verucosa (Mandabam)>G. salicornia (Mandabam)>G. ferugosonii (Tuticorin)>G. ferugosonii (Mandabam)>G. edulis (Pulicate)>G. corticata (Tuticorin)>G. verucosa (Pulicate)>G. corticata var. cylindrica (Tuticorin).Conclusion: The results of the present study clearly explained the anti-inflammatory potential of the studied Gracillaria species. The aqueous 0extracts of Gracillaria collected from Mandapam showed the highest percentage of protection activity. This study results confirm the existence of active principle responsible for the anti-inflammatory activity. Further studies are required to isolate the active principles without any side effects

Intracellular granzyme A expression of peripheral blood lymphocyte subsets in pulmonary tuberculosis

Cell-mediated immunity is a key weapon of host defence against tuberculosis (TB). Granzyme A (GzmA), a serine protease, present in the granules of cytotoxic cells induces caspase-independent cell death. We estimated the proportion of GzmA producing lymphocyte subsets in peripheral blood from 59 normal healthy volunteers and 48 pulmonary TB (PTB) patients using flow cytometry. When compared with normal healthy subjects, we observed a significantly higher percentage of GzmA-positive CD56+ cells (P = 0.01) in PTB patients. However, when the absolute number was compared between the two groups, a significantly decreased number of GzmA-expressing CD16+ (P = 0.01) and CD56+ (P = 0.0001) cells was observed in patients and this could be explained by the significantly reduced number of total lymphocytes (P = 0.0009) seen in the patients. There was no significant difference in the number of CD4+ and CD8+ GzmA double-positive cells between the two study groups. CD56 is a natural killer cell marker and these cells represent innate immune response to TB. We report an increased percentage of CD56+ cells expressing GzmA in TB patients, which shows the relevance of the GzmA-mediated pathway of apoptosis in immunity against Mycobacterium tuberculosis

Interferon gamma (IFN -gamma) and interleukin -4 (IL-4) gene variants and cytokine levels in pulmonary tuberculosis. Indian Journal of Medical Research

Background & objectives: Cytokine gene polymorphisms may alter Th1/Th2 balance with major implications in tuberculosis. The aim of our study was to find out whether Interferon � +874A and IL-4 -590T polymorphisms were associated with susceptibility to pulmonary tuberculosis as well as the level of IFN� and IL-4 in south Indian population. Methods: Interferon � +874A and IL-4 -590T promoter polymorphisms were studied in 129 pulmonary tuberculosis (PTB) patients and 127 normal healthy subjects (NHS) and were associated with culture filtrate and live Mycobacterium tuberculosis induced IFN� and IL-4 production in peripheral blood mononuclear cells (PBMCs). IL-4 gene variants were also associated with IgG antibody levels against M. tuberculosis culture filtrate antigen. Results: The variant IFN� genotypes and IFN� levels between genotypes did not differ significantly in patients and controls. Significantly increased frequency of variant IL-4 ‘CT’ genotype in PTB patients (P<0.05) and ‘CC’ genotype in control group (P<0.01) was observed. IL-4 levels were detectable in very few subjects and the IgG levels did not differ between the three IL-4 genotypes. Interpretation & conclusion: The study suggests a lack of functional association of Interferon � +874A polymorphism in tuberculosis in south Indian population. The higher frequency of IL-4 ‘CT’ genotype in PTB suggests a possible association of IL-4 -590T promoter polymorphism with susceptibility to tuberculosis, and the ‘CC’ genotype may be associated with protection

Elevated percentage of perforin positive cells in active pulmonary tuberculosis

Background & objectives: Perforin is one of the major effector molecules of cytotoxic cells associated with killing of cells harbouring intracellular bacterial infection. The precise role of perforin positive cells in tuberculosis still remains controversial. The present study was done to determine the number of circulating CD4+ and CD8+ perforin positive cells to assess the level of cytotoxic response against Mycobacterium tuberculosis in patients with pulmonary tuberculosis. Methods: Intracellular perforin and surface CD4 and CD8 staining of peripheral blood lymphocytes was done using specific monoclonal antibodies and enumerated using flowcytometry. Results: A significantly decreased total lymphocytes (P<0.01), CD4 (P<0.001) and CD8 (P<0.01) lymphocyte counts in PTB patients was observed compared to normal healthy individuals (NHS). Intracellular perforin staining showed significantly elevated percentages of total (P<0.05) and CD8 (P<0.01) perforin positive cells in PTB patients compared to NHS. However, the absolute counts of total, CD4 and CD8 cells positive for perforin were similar in patients and NHS. Interpretation & conclusion: Our results suggest that during active stage of pulmonary tuberculosis there was an increased percentage of CD8 cells positive for perforin, irrespective of their absolute counts. Further, CD8+ perforin positive cells ma

Differential Expression of Cytokines in Response to Respiratory Syncytial Virus Infection of Calves with High or Low Circulating 25-Hydroxyvitamin D3

Deficiency of serum levels of 25-hydroxyvitamin D3 has been related to increased risk of lower respiratory tract infections in children. Respiratory syncytial virus (RSV) is a leading cause of low respiratory tract infections in infants and young children. The neonatal calf model of RSV infection shares many features in common with RSV infection in infants and children. In the present study, we hypothesized that calves with low circulating levels of 25-hydroxyvitamin D3 (25(OH)D3) would be more susceptible to RSV infection than calves with high circulating levels of 25(OH)D3. Calves were fed milk replacer diets with different levels of vitamin D for a 10 wk period to establish two treatment groups, one with high (177 ng/ml) and one with low (32.5 ng/ml) circulating 25(OH)D3. Animals were experimentally infected via aerosol challenge with RSV. Data on circulating 25(OH)D3 levels showed that high and low concentrations of 25(OH)D3 were maintained during infection. At necropsy, lung lesions due to RSV were similar in the two vitamin D treatment groups. We show for the first time that RSV infection activates the vitamin D intracrine pathway in the inflamed lung. Importantly, however, we observed that cytokines frequently inhibited by this pathway in vitro are, in fact, either significantly upregulated (IL-12p40) or unaffected (IFN-γ) in the lungs of RSV-infected calves with high circulating levels of 25(OH)D3. Our data indicate that while vitamin D does have an immunomodulatory role during RSV infection, there was no significant impact on pathogenesis during the early phases of RSV infection. Further examination of the potential effects of vitamin D status on RSV disease resolution will require longer-term studies with immunologically sufficient and deficient vitamin D levels

Use of open access and licensed e-books by the researchers in academic and research institutes in Karnataka

The electronic books are among most extensively used electronic resources and its impact on academic community is much important. There are several managerial and technical prospects that affect usability of electronic books by research scholars in an academic institution. The purpose ofthis study is to investigated the use of open access and licensed electronic books by the research scholars in academic and research institutionsin Karnataka. The population of the study consists ofresponses from1110research scholars.The survey method was adoptedusing questionnaire as a tool for data collections. It is clear from the study that large number of research scholars are aware and use electronic books for their academic and research purpose. The institutes should subscribe large number of e-books for target users and proper training should be given to maximise the use of e-books

1, 25 Dihydroxyvitamin D3 modulated cytokine response in pulmonary tuberculosis

1, 25 Dihydroxyvitamin D3 (1, 25(OH)2 D3) has gained significant importance in tuberculosis with regard to its immunoregulatory activities. Our aim was to evaluate the effect of 1, 25(OH)2 D3 on cytokine response to Mycobacterium tuberculosis antigens in pulmonary tuberculosis. Peripheral blood mononuclear cells obtained from 60 healthy controls and 52 pulmonary tuberculosis patients were cultured with culture filtrate antigen (CFA) of M. tuberculosis and live M. tuberculosis with and without 1, 25(OH)2 D3 (10�9, 10�8 and 10�7 M concentrations). The culture supernatants were used to estimate IL-8, IL-6, TGF-b, IL-10, IFN-c, IL-12p40, IL-2, IL-4 and IL-5 levels by ELISA. 1, 25 Dihydroxyvitamin D3 significantly suppressed IL-12p40 and IFN-c production in response to CFA and live M. tuberculosis with a maximum suppression at 10�7M concentration (p < 0.0001). In CFA stimulated cultures, addition of 1, 25(OH)2 D3 significantly suppressed IL-8, IL-6 and IL-10 whereas the IL-2 levels were significantly increased in controls. It variably influenced the Th2 cytokines, showing an increased trend for IL-4 and suppressed IL-5 levels. We report that 1, 25(OH)2 D3 differentially modulates production of cytokines in response to M. tuberculosis antigens by predominantly suppressing IL-12p40 and IFN-c production in a dose dependent manner. Our results suggest a role for vitamin D in restricting acquired immune response against tuberculosis by regulating cytokine production

Regulatory role of 1, 25-dihydroxyvitamin D3 and vitamin D receptor gene variants on intracellular granzyme A expression in pulmonary tuberculosis

Vitamin D receptor (VDR) genotypes have been shown to be associated with differential susceptibility or resistance to tuberculosis. The influence of FokI, BsmI, ApaI and TaqI variants of VDR gene on 1, 25(OH)2 D3 modulated granzyme A expression of cytotoxic lymphocytes induced by culture filtrate antigen (CFA) of Mycobacterium tuberculosis was studied in 40 pulmonary tuberculosis (PTB) patients and 49 normal healthy subjects (NHS) by flow cytometry. In both the study groups, addition of 1, 25(OH)2 D3 (10−7M) significantly reduced the percentage of granzyme A positive cells in both unstimulated (NHS, pb0.0001; PTB, p=0.02) and stimulated culture conditions (CFA, NHS, pb0.0001; PTB, p=0.0001) which correlated positively with the IFN-γ levels (unstimulated, p=0.01; CFA stimulated, p=0.004) in NHS. The ApaI aa genotype and bbaaTT extended genotype were associated with a significantly decreased percentage of granzyme A positive cells in NHS (pb0.05). Our results suggest that 1, 25(OH)2 D3 suppresses granzyme A probably by down-regulating Th1 cytokine response. Moreover, the VDR gene variants might regulate cytotoxic T-cell response via 1, 25 (OH)2 D3 mediated suppression of granzyme A expression in tuberculosis

Regulatory region polymorphisms of vitamin D receptor gene in pulmonary tuberculosis patients and normal healthy subjects of south India

Vitamin D receptor (VDR) gene variants are associated with differential susceptibility or resistance to tuberculosis in different ethnic groups. We investigated the polymorphisms in the 5′ regulatory region of VDR gene in 206 normal healthy subjects and 166 patients with pulmonary tuberculosis from south India. Cdx-2 polymorphism was studied by polymerase chain reaction (PCR) with allelespecific primers, while genotyping of A1012G was done by PCR-based restriction fragment length polymorphism. A significantly decreased frequency of Cdx-2 G allele (P = 0.016) and G/G genotype (P = 0.010) and an increased frequency of A-A haplotype (A allele of Cdx-2 and A allele of A1012G) (P = 0.015) were observed in patients compared to controls. The study suggests that Cdx-2 G/G genotype may be associated with protection and A-A haplotype with susceptibility to tuberculosis