Epidemiological studies of hepatitis B: Preliminary report on adjuvant potential of Calotropis gigantea and Ficus religiosa against hepatitis B vaccine containing surface antigen

Background and aims: The frequency of chronic hepatitis B virus infection varies totally in different part of the world but it could be categorized as high, intermediate and low endimicity. In this regard, development of safe and effective vaccines against infectious diseases especially hepatitis B virus that are still required for emerging new pathogens, re-emerging old pathogens and in order to improve the inadequate protection conferred by existing vaccines. In this study, immunological studies were conducted pertaining to investigate the aqueous leaves extract of Calotropis gigantea and Ficus religiosa for determining its adjuvant effect against hepatitis B vaccine containing surface antigen (HBsAg). Methods: In this study, our group evaluated the secondary metabolites that are present in the aqueous extract using high performance thin layer chromatography (HPTLC) and estimated its antibody (IgG) titre against HBsAg using variable doses (2.5-10 mg) of aqueous leaves extract of these medicinal plants and also determined splenocyte proliferation assay (ex vivo studies) on day 4 where mice (n=5) were immunized subcutaneously on day 0 with HBsAg (20 µg/ml, 100 µl). Results: The results showed that aqueous leaves extract showed anti-HBsAg titre and also enhanced splenocyte proliferation assay at higher doses (10 mg, 50 µl) as compared to control. Conclusion: Overall, the results showed that aqueous leaves extract of Calotropis gigantea and Ficus religiosa showed adjuvant activity against HBsAg

<p style="margin: 0cm 0cm 8pt; text-align: center; line-height: normal;">Virucidal potential of Prosopis spicigera and Mangifera indica on human peripheral blood mononuclear cells</p>

Introduction: The treatment of viral infections with the available antiviral drugs is not free of side effects. Therefore, in the present study, our group focused on antiviral activity against Newcastle disease (NDV) and IBD viruses using medicinal plants especially leaves of Prosopis spicigera and Mangifera indica.Methods: Different medicinal plant products especially leaves of P. spicigera and M. indica were tested in the form of aqueous leaves extracts (0.5- 30 mg/mL; 50 &mu;L) for anti-microbial activities on human peripheral blood mononuclear cells (PBMC) pertaining to determine their proliferation rate (cytotoxicity assay), tumor necrosis factor alpha (TNF&alpha;) production and CD14 monocyte surface marker.Results: Three medicinal plant aqueous extracts showed significant antimicrobial activity against PBMC at higher doses with respect to decline in proliferation assay, TNF&alpha; production and CD14 monocyte surface marker as compared to control.Conclusion: Aqueous leaves extract of P. spicigera and M. indica showed antimicrobial activities and might be useful for the treatment of various viral diseases.</p

Flow cytometric evaluation of Calotropis gigantea for determining its antimicrobial activities in infected human whole blood samples

Introduction: Medicinal plants are considered to be safer, non-toxic and less harmful as compared to synthetic based drugs that are available. In this study, we focused on aqueous leaves extract of Calotropis gigantea for determining its antimicrobial activity in infected (dengue) human whole blood samples using flow cytometry.Methods: Infected dengue human blood samples (n = 5; confirmed on the basis of NS1 antigen to dengue virus;) were collected from pathology lab and evaluated its blood counts (lymphocytes, monocytes and granulocytes count); forward scatter (FSC) and side scatter (SSC) including CD14 monocyte surface marker following the use of variable doses of aqueous leaves extract of C. gigantea.Results: In this study, the results showed that aqueous leaves extract of C. gigantea caused enhancement in case of granulocytes FSC (shape and size) and SSC (granularity) counts but this aqueous extract inhibited CD14 monocyte surface marker population at higher doses. In contrast, dengue infected human blood samples used as control showed sudden decline in granulocytes count but there was enhancement in CD14 monocyte surface marker as compared to control group.Conclusion: Overall, C. gigantea in the form of aqueous leaves extract showed anti-dengue activity in infected human whole blood samples.</p

Immunosuppressive activity of terpenoids from Mesua ferrea, Ficus benghalensis and Butea frondosa on human whole blood

Background and aims: Terpenoids are one of the largest groups of natural compounds and is generally used for the treatment of human diseases e.g. taxol (anticancer drug) and artimesinin (antimalarial drug). Thus, these agents (terpenoids) derived from medicinal plant products that are effective against intracellular or extracellular infections and these are urgently required. Therefore, this research was directed to assess its immunosuppressive activity of crude terpenoids on hepatitis B vaccine containing surface antigen (HBsAg) on human whole blood. Methods: In this study, crude terpenoids (6.25-25 mg/ml; 50 µl) extricate from the leaves of these medicinal plants and evaluated its immunosuppressive activity on HBsAg (20 µg/ml) using human whole blood in order to determine the analysis of blood counts (flow cytometry), cytotoxicity assay (MTT (3-4,5-dimethylthiazol–2-yl-2, 5-diphenyltetrazolium bromide) and nitric oxide (NO) production from cell culture supernatant. Results: The results showed that these terpenoids showed drastic decrease in monocytes and granulocytes count but increased in lymphocytes count and inhibited NO production at higher doses (25 mg/ml; 50 µl). Conclusion: These terpenoids exhibited immunosuppressive activity and could be a promising source of medicinally important natural compound

A longitudinal study of students\u27 reasoning about variation in distributions in an introductory college statistics course

Current curricular documents including the Common Core State Standards (2010) and the Guidelines for Assessment and Instruction in Statistics Education (2005) have increased the need for students\u27 understanding and reasoning about statistics at both the K-12 and college levels. In addition, an increasing number of students are taking the Advanced Placement Statistics Exam (College Board, 2011) or a college-level introductory statistics course (Scheaffer & Stasny, 2000). One of the main components for statistical thinking is consideration of variation (Wild & Pfannkuch, 1999). Previous studies have shown that students have misconceptions about variation (e.g. Reading, 2004; Torok & Watson, 1999) and students often lack the ability to give sophisticated answers (Shaughnessy, 2007). The goal of this study was to better understand how students\u27 reasoning about variation in a distributional context changes as they progress through an introductory college-level statistics course. In order to better understand the longitudinal nature of this process during a semester-long introductory statistics course, both quantitative and qualitative data were collected at three different times (beginning, middle, and end of the course) in surveys and interviews. The Structure of Observed Learning Outcomes (SOLO) Taxonomy (Biggs & Collis, 1982) was used to understand and assess the quality of their reasoning. Qualitative data came from two sources: three interviews from each of the ten interviewees and three survey questions on each of three surveys from all participants. The interviews were transcribed and responses were sorted into appropriate locations in the SOLO Taxonomy. After coding responses to each question in each interview, themes of progress were then identified. These themes showed that students progressed through four different paths of reasoning including: improved, maintained, decreased, and inconsistent. Quantitative data showed that while students were good at reasoning about situations involving bar graphs and dot plots with regards to comparing variability in distributions, they struggled with reasoning about histograms. Overall, this study found that there was no statistically significant improvement in reasoning about variability when comparing distributions as students progressed through a college-level introductory statistics course. This lack of improvement suggested that perhaps college students needed to have direct intervention or cognitive conflict in order to make more progress in reasoning about variability when comparing distributions

Potential immunosuppressive and anti-inflammatory activity of aqueous extract of Mangifera indica

Background: There is a worldwide interest in searching for potential and effective medicinal plant candidates against various diseases or disorders. Till now, there are number of anti-inflammatory drugs like NSAIDs are available which showed various adverse effects in our body. To reduce these adverse effects, some of medicinal plants have been experimentally validated. Aim: The aim of our immunopharmacological study is to investigate the immunosuppressive and anti-inflammatory activity of aqueous leaves extract of Mangifera indica in human peripheral blood mononuclear cells (PBMC) extracellular against hepatitis B vaccine antigen (HBsAg) Methods: In this study, aqueous leaves extract were collected from Mangifera indica and determined its effect on total blood counts (using forward and side scatter) and monocyte CD14 FITC surface markers using flow cytometry. In addition, the effect of aqueous leaves extract on nitric oxide (NO) production from PBMC cell culture supernatant and also estimate its proliferation assay using Concanavalin A (Con A, 0.5 mg/ml, 50 µl). Data analysis was performed using BD cell Quest Pro software for flow cytometric analysis and one way ANOVA test (Boniferroni multiple comparison test). Results: The results displayed that aqueous leaves extract of Mangifera indica showed dose dependent decline in blood counts (increased in side scatter and slightly decreased in forward scatter), monocyte CD14 FITC surface marker; Con A proliferation and nitric oxide production from cell culture supernatant in human PBMC. Conclusion: Overall, the results claimed that aqueous leaves extract of Mangifera indica at higher doses showed immunosuppressive and anti-inflammatory activity

Immunopharmacological screening of aqueous root extract of Santalum album

Introduction: In order to scrutinize the anti-microbial and anti-inf lammatory agents, manyresearch groups have focused their works on the field of immunopharmacology. Thus, the currentstudy aimed to evaluate the immunopharmacological activity of aqueous root extract of Santalumalbum against Hepatitis B surface antigen (HBsAg) and Newcastle disease virus (NDV) on humanperipheral blood mononuclear cells (PBMCs).&nbsp;Methods: Variable doses of aqueous root extract of Santalum album (0.5-30 mg/ml, 50 &micro;l) were&nbsp;prepared using phosphate buffered saline (PBS) and tested for proliferation assay, nitric oxideproduction and CD14 monocyte surface marker using flow cytometry.Results: Aqueous root extract of Santalum album inhibited proliferation, nitric oxide productionand CD14 monocyte surface marker at higher doses.Conclusion: Aqueous root extract of Santalum album possesses both anti-inf lammatory as well asantiviral activities and might be used for these purposes.&nbsp

Anti-inflammatory and anti-microbial activities of aqueous leaves extract of Butea frondosa

Introduction: Butea frondosa has been suggested to be very useful in treating inflammatory diseases but no scientific investigation has been done in such direction. In this study the anti-inflammatory and anti-microbial activities of leaves aqueous extract of B. frondosa were determined in infected and non-infected human whole blood against specific vaccine antigen, HBsAg.Methods: In order to explore the anti-inflammatory and anti-microbial activities of B. frondosa (0.5&ndash;0 mg/mL; 50 &mu;l), infected (virally) and non infected (control) human whole blood samples were stimulated with hepatitis B vaccine containing surface antigen (HBsAg, 20 &mu;g/mL;10 &mu;l) in order to determined its blood counts and proliferation assay.Results: Aqueous leaves extract of B. frondosa (10 mg/mL; 50 &mu;l) containing HBsAg inhibited the percentage count of monocytes as well as granulocytes population in both cases. In addition, this aqueous extract also reduced its proliferation rate at higher doses.Conclusion: Aqueous leaves extract of B. frondosa possesses both anti-inflammatory and antimicrobial activities and might be used for these purposes

Haemolytic and immunoadjuvant effect of Butea frondosa on the immune response to hepatitis B vaccine containing surface antigen in mice

Introduction: Immunological adjuvants derived from various synthetic micro-organisms or from medicinal plant products enhance specific immune responses against vaccine antigens. Immunological studies have already done pertaining to identify compounds from medicinal plant metabolites that are suitable for vaccine formulation. In this study, aqueous leave extracts of Butea frondosa were selected to evaluate their haemolytic activity and immunoadjuvant effects.Methods: For this study, immunoadjuvant activity was investigated ex vivo in mice model based studies using splenocyte proliferation assay and also measured IgG titre in cell culture supernatant (using indirect ELISA). Swiss mice were immunized subcutaneously with specific protein antigen i.e. hepatitis B vaccine containing surface antigen (HBsAg, 20 &mu;g/mL; 10 &mu;l) on day 0 and collected the spleen cells on day 4 and proceeded for proliferation assay of variable doses of aqueous leaves extracts of Butea frondosa (0.5&ndash;30 mg/ml; 50 &mu;l) along with hepatitis B surface antigen (HBsAg) (challenging dose; 20 &mu;g/mL; 10 &mu;l) and also estimated the antibody (IgG) titre in splenocyte cell culture supernatant including determination of its haemolytic activity in human whole blood samples.Results: The results demonstrated that aqueous leave extracts stimulated HBsAg population at lower doses (0.5 mg/mL) and also enhanced IgG titre as compared with control and HBsAg treated group. In addition, aqueous leaves extracts of Butea frondosa showed anti-HBsAg titre at higher doses and also showed slightly haemolytic effect in human whole blood.Conclusion: These results suggested that aqueous extract of Butea frondosa may represent viable candidate for effective vaccine adjuvants due to their higher immune response (with respect to IgG titre and proliferation assay) and lower or non-haemolytic effects.</p

Aqueous extract of Calamus rotang as a novel immunoadjuvant enhances both humoral and cell mediated immune response

Introduction: Search for new adjuvants for human vaccines has become an expanding field of research in the last thirty years for generating stronger vaccines, capable of inducing protective and long lasting immunity in humans. The objective of this study was to investigate the immunoadjuvant activity of aqueous extract from the leaves of Calamus rotang using phosphate buffered saline (PBS, pH 7.2) against hepatitis B vaccine containing surface antigen (HBsAg; 20 &mu;g/mL).Methods: In this research qualitative study was evaluated in order to determine the presence of secondary metabolites and further confirmation of these metabolites through high performance thin layer chromatography (HPTLC) and identification by liquid chromatography mass spectrometry (LC-MS). In addition, indirect Elisa was performed using HBsAg as coating antigen and this aqueous extract showed anti-HBsAg titre at higher doses as compared to standard and control. In continuation of these studies, Swiss mice were immunized subcutaneously on day 0 with HBsAg (20 &mu;g/mL, 100 &mu;L) and collect splenocytes on day 4 for splenocyte proliferation assay (ex vivo studies; again exposed with HBsAg) and estimation of Th1 (IFN-gamma and tumor necrosis factor TNF-&alpha;) cytokines from cell culture supernatant.Results: The aqueous leaves extract of C. rotang showed dose dependent enhancement in antibody titre and proliferation at higher doses (P &lt; 0.01) with respect to HBsAg. In addition, this aqueous extract also showed improvement in Th1 (IFN-gamma and TNF alpha) cytokines at higher doses (P &lt; 0.01) from cell culture supernatant as compared to standard HBsAg.Conclusion: Calamus rotang has additive adjuvant activity against hepatitis B vaccine antigen containing alum and may help to raise antibodies against HBsAg under challenging administration regimen and might be a potent vaccine adjuvant.</p