Protein arginine deiminase (PAD) enzymes catalyze the conversion of protein-bound arginine into c... more Protein arginine deiminase (PAD) enzymes catalyze the conversion of protein-bound arginine into citrulline, an irreversible posttranslational modification with loss of a positive charge that can influence protein–protein interactions and protein structure. Protein arginine deiminase activity depends on high intracellular calcium concentrations occurring in dying cells. In this study, we demonstrate that protein citrullination is common during pyroptotic cell death in macrophages and that inhibition of PAD enzyme activity by Cl-amidine, a pan-PAD inhibitor, blocks NLRP3 inflammasome assembly and proinflammatory IL-1β release in macrophages. Genetic deficiency of either PAD2 or PAD4 alone in murine macrophages does not impair IL-1β release; however, pharmacological inhibition or small interfering RNA knockdown of PAD2 within PAD4−/− macrophages does. Our results suggest that PAD2 and 4 activity in macrophages is required for optimal inflammasome assembly and IL-1β release, a finding o...
The CD1d-restricted invariant natural killer T (iNKT) cells are implicated in innate immune respo... more The CD1d-restricted invariant natural killer T (iNKT) cells are implicated in innate immune responses against human immunodeficiency virus (HIV). However, the determinants of cellular dysfunction across the iNKT cells subsets are seldom defined in HIV disease. Herein, we provide evidence for the involvement of the negative checkpoint regulator (NCR) 2B4 in iNKT cell alteration in a well-defined cohort of HIV-seropositive anti-retroviral therapy (ART) naïve, ART-treated, and elite controllers (ECs). We report on exaggerated 2B4 expression on iNKT cells of HIV-infected treatment-naïve individuals. In sharp contrast to CD4(-)iNKT cells, 2B4 expression was significantly higher on CD4(+) iNKT cell subset. Notably, an increased level of 2B4 on iNKT cells was strongly correlated with parameters associated with HIV disease progression. Further, iNKT cells from ART-naïve individuals were defective in their ability to produce intracellular IFN-γ. Together, our results suggest that the levels ...
International Archives of Allergy and Immunology, 2016
Aim of Study: We used a triplex real-time polymerase chain reaction (PCR) to classify our common ... more Aim of Study: We used a triplex real-time polymerase chain reaction (PCR) to classify our common variable immunodeficiency (CVID) patients into distinct groups according to the amount of their T-cell receptor excision circles (TRECs) and κ-deleting recombination excision circles (KRECs). Materials and Methods: TREC and KREC analysis was performed using a multiplex real-time PCR assay. The T- and B-lymphocyte subsets were measured by flow cytometry. Results: The copy number of TRECs and KRECs was significantly reduced in CVID patients compared to healthy controls. The TREC copy number was inversely correlated with age in both healthy subjects and patients; however, the KREC copy number was inversely correlated with age only in CVID patients. Moreover, no association was seen between TREC/KREC copy number and clinical manifestations such as bronchiectasis, splenomegaly, granulomata, autoimmune cytopenias, organ-specific autoimmunity, enteropathy and lymphoid hyperplasia. Conclusion: T...
Summary Immune senescence as well as disturbed CD8+ T cell differentiation are a hallmark of chro... more Summary Immune senescence as well as disturbed CD8+ T cell differentiation are a hallmark of chronic HIV infection. Here, we investigated to what extent immune senescence is reversible after initiation of anti-retroviral treatment (ART). Peripheral blood mononuclear cells (PBMCs) from a cohort of HIV patients with different disease courses, including untreated viral controllers (n = 10), viral non-controllers (n = 16) and patients on ART (n = 20), were analysed and compared to uninfected controls (n = 25) by flow cytometry on bulk and HIV-specific major histocompatibility complex (MHC) class I tetramer+ CD8+ T cells for expression of the memory markers CCR7 and CD45RO, as well as the senescence marker CD57 and the differentiation and survival marker CD127. Furthermore, a subset of patients was analysed longitudinally before and after initiation of ART. Frequencies of CD57+CD8+ T cells decreased after initiation of ART in central memory (Tcm) but not in effector memory T cell populat...
To function optimally, human blood natural killer (NK) cells need to communicate with other immun... more To function optimally, human blood natural killer (NK) cells need to communicate with other immune cells. Previously, it has been shown that NK cells communicate with 6-sulfo LacNAc dendritic cells (slanDCs), which are able to stimulate NK cells in vitro. In this study, we investigated how slanDCs regulate the level of NK cell activation. The secretion of interleukin (IL)-1β by slanDCs during coculture with NK cells increased as a result of signaling via intercellular adhesion molecule-1 on slanDCs following its interaction with lymphocyte function-associated antigen-1 on NK cells. IL-1β induced the expression of Fas receptor (CD95) on NK cells. The binding of Fas ligand (CD178) to CD95 induced the apoptosis of activated NK cells. Moreover, IL-1β also induced increased cyclooxygenase-2 expression in slanDCs, which in turn enabled the cells to secrete prostaglandin (PG)-E2. Consequently, PGE2 acted as a suppressing agent, tuning down the activation level of NK cells. In summary, IL-1...
HIV-HCV co-infection is associated with accelerated progression to hepatic fibrosis, cirrhosis an... more HIV-HCV co-infection is associated with accelerated progression to hepatic fibrosis, cirrhosis and hepatocellular carcinoma than HCV mono-infection. The contribution of innate immunity during HIV-HCV co-infection has been a relatively under-investigated area. Natural killer (NK) cells are pivotal sentinels of innate immunity against viruses and tumour cells. In this study we evaluated the effect of HIV-HCV co-infection on peripheral blood NK cell subsets with emphasis on the phenotype of CD56(bright) NK cells. Sixty patients were included in the study; HIV mono-infected (n = 12), HCV mono-infected (n = 15), HCV-HIV co-infected (n = 21) and healthy controls (n = 16). PBMCs were isolated and immunophenotyping of NK cells was performed by flowcytometry. We observed an expansion of CD56(bright) NK cell subset in HIV-HCV co-infection as compared to healthy controls and HIV mono-infected group. All the infected groups had an upregulated expression of the activating receptor NKG2D on CD56(...
Cellular and molecular life sciences : CMLS, Jan 5, 2015
Natural killer (NK) cells are innate immune effectors that provide first line of defence against ... more Natural killer (NK) cells are innate immune effectors that provide first line of defence against viruses. Human NK cells are heterogeneous in nature, and their functions rely on a dynamic balance between germ-line-encoded activating and inhibitory receptors. HIV-1 infection results in altered NK cell receptor repertoire and impaired effector functions including the ability to lyse virus-infected cells and secretion of antiviral cytokine IFN-γ. Over the last decade, additional NK cell subset-specific molecules have been identified, leading to emergence of a more complex cellular diversity than previously thought. Herein, we discuss NK cell subset redistribution, altered receptor repertoire and influence of interaction of polymorphic leucocyte antigen (HLA) and killer cell immunoglobulin-like receptors (KIR) on HIV-1 disease progression.
Natural killer (NK) cells are effector and regulatory innate immune cells and play a critical rol... more Natural killer (NK) cells are effector and regulatory innate immune cells and play a critical role in the first line of defense against various viral infections. Although previous reports have indicated the vital contributions of NK cells to HIV-1 immune control, nongenetic NK cell parameters directly associated with slower disease progression have not been defined yet. In a longitudinal, retrospective study of 117 untreated HIV-infected subjects, we show that higher frequencies as well as the absolute numbers of CD8(+) CD3(-) lymphocytes are linked to delayed HIV-1 disease progression. We show that the majority of these cells are well-described blood NK cells. In a subsequent cross-sectional study, we demonstrate a significant loss of CD8(+) NK cells in untreated HIV-infected individuals, which correlated with HIV loads and inversely correlated with CD4(+) T cell counts. CD8(+) NK cells had modestly higher frequencies of CD57-expressing cells than CD8(-) cells, but CD8(+) and CD8(-...
NK cells are pivotal sentinels of the innate immune system and distinct subpopulations in periphe... more NK cells are pivotal sentinels of the innate immune system and distinct subpopulations in peripheral blood have been described. A number of studies addressed HIV-induced alterations of NK cell phenotype and functionality mainly focusing on CD56 dim CD16 + and CD56 2 CD16 + NK cells. However, the impact of HIV-infection on CD56 bright NK cells is less well understood. Here we report a rise of CD56 bright NK cells in HIV-infected individuals, which lack CCR7-expression and strongly correlate with HIV viral load. CCR7 2 CD56 bright NK cells were characterized by increased cytolytic potential, higher activation states and a more differentiated phenotype. These cells thus acquired a number of features of CD56 dim CD16 + NK cells. Furthermore, CD56 bright NK cells from HIV patients exhibited higher degranulation levels compared to uninfected individuals. Thus, chronic HIV-infection is associated with a phenotypic and functional shift of CD56 bright NK cells, which provides a novel aspect of HIV-associated pathogenesis within the NK cell compartment.
Coxiella burnetii (Cb) is a hardy, stealth bacterial pathogen lethal for humans and animals. Its ... more Coxiella burnetii (Cb) is a hardy, stealth bacterial pathogen lethal for humans and animals. Its tremendous resistance to the environment, ease of propagation, and incredibly low infectious dosage make it an attractive organism for biowarfare. Current research on the classification of Coxiella and features influencing its presence in the soil is generally confined to statistical techniques. Machine learning other than traditional approaches can help us better predict epidemiological modeling for this soil-based pathogen of public significance. We developed a two-phase feature-ranking technique for the pathogen on a new soil feature dataset. The feature ranking applies methods such as ReliefF (RLF), OneR (ONR), and correlation (CR) for the first phase and a combination of techniques utilizing weighted scores to determine the final soil attribute ranks in the second phase. Different classification methods such as Support Vector Machine (SVM), Linear Discriminant Analysis (LDA), Logist...
The aim of this study is to analyse the influence of LILRA3 and the genetic leukocyte immunoglobu... more The aim of this study is to analyse the influence of LILRA3 and the genetic leukocyte immunoglobulin-like receptor 3 (LILRA3) deletion on transmission and clinical course of HIV infection. Case and control study. LILRA3 genotypes were determined by PCR. HIV patients were categorized into short-term progressors, normal progressors and long-term nonprogressors according to the clinical course. Functional studies were performed using real-time PCR, intracellular flow cytometry and ELISA. The prevalence of the homozygous LILRA3 deletion was higher in HIV-positive individuals (n = 439) than in controls (n = 651) (P = 0.02). The disease progression was faster in homozygously deleted patients with more short-term progressors than in heterozygous (P = 0.03) and homozygously positive (P = 0.002) individuals. These results have been confirmed in a seroconverter cohort (n = 288). The frequency of the homozygous deletion in the confirmation cohort was higher than in controls (P = 0.04). Combini...
Human natural killer (NK) cells are one major component of lymphocytes that mediate early protect... more Human natural killer (NK) cells are one major component of lymphocytes that mediate early protection against viruses and tumor cells, and play an important role in immune regulatory functions. In this study, we demonstrated that human NK cells could be divided into four subsets, CD56 hi CD16 À , CD56 lo CD16 À , CD56 þ CD16 þ and CD56 À CD16 þ , based on the expression of cell surface CD56 and CD16 molecules. Phenotypic analysis of NK cell subsets indicated that the expression of activation markers, adhesion molecules, memory cell markers, inhibitory and activating receptors, and intracellular proteins (granzyme B and perforin) were heterogeneous. Following interleukin (IL)-2 stimulation, interferon-g was preferentially produced by CD56 þ CD16 À NK cells and this subset showed more proliferative capacity. The cytolytic activity of both CD56 þ CD16 À and CD56 þ/À CD16 þ subsets could be augmented in response to IL-2. The data provided a new definition for NK cell subsets demonstrating their phenotypic and functional diversity and possible stage of NK cell differentiation in peripheral blood.
Cooperative learning is being used throughout the world. It is gaining international popularity. ... more Cooperative learning is being used throughout the world. It is gaining international popularity. On the basis of theory and practice, cooperative learning methods have been used and tested in different cultures and classroom situations. Therefore, a study has been conducted in order to examine the "comparative effectiveness of cooperative learning and traditional approaches of teaching Islamic Studies at Elementary level in Pakistan". Pretest posttest experimental design was used. Students of class six were equally divided into two groups on the basis of teacher made pretest scores. As a treatment cooperative learning method Student-Teams-Achievement-Division (STAD) was provided to experimental group, while control group was taught by using traditional methods of teaching, and kept under traditional classroom situation for the period of 12 weeks. At the end of the treatment, teacher made posttest was administered to measure the achievement scores of students. Significance of difference between the mean scores of both groups was tested by applying t-test at 0.05 level of significance. Findings of the study revealed that both the groups were equal in the beginning of the experiment. The experimental group showed better results on posttest scores, showing the obvious preeminence of cooperative learning method over traditional methods of teaching. High and low achievers of experimental group showed better result as compared to control group, and also helpful in developing creative thinking. Results of the study indicate that cooperative learning is more effective teaching method for Islamic Studies as compared to traditional methods. It revealed that it was more 2 REVIEW OF THE RELATED LITERATURE 9 xvi APPENDICES Appendix Topic
Protein arginine deiminase (PAD) enzymes catalyze the conversion of protein-bound arginine into c... more Protein arginine deiminase (PAD) enzymes catalyze the conversion of protein-bound arginine into citrulline, an irreversible posttranslational modification with loss of a positive charge that can influence protein–protein interactions and protein structure. Protein arginine deiminase activity depends on high intracellular calcium concentrations occurring in dying cells. In this study, we demonstrate that protein citrullination is common during pyroptotic cell death in macrophages and that inhibition of PAD enzyme activity by Cl-amidine, a pan-PAD inhibitor, blocks NLRP3 inflammasome assembly and proinflammatory IL-1β release in macrophages. Genetic deficiency of either PAD2 or PAD4 alone in murine macrophages does not impair IL-1β release; however, pharmacological inhibition or small interfering RNA knockdown of PAD2 within PAD4−/− macrophages does. Our results suggest that PAD2 and 4 activity in macrophages is required for optimal inflammasome assembly and IL-1β release, a finding o...
The CD1d-restricted invariant natural killer T (iNKT) cells are implicated in innate immune respo... more The CD1d-restricted invariant natural killer T (iNKT) cells are implicated in innate immune responses against human immunodeficiency virus (HIV). However, the determinants of cellular dysfunction across the iNKT cells subsets are seldom defined in HIV disease. Herein, we provide evidence for the involvement of the negative checkpoint regulator (NCR) 2B4 in iNKT cell alteration in a well-defined cohort of HIV-seropositive anti-retroviral therapy (ART) naïve, ART-treated, and elite controllers (ECs). We report on exaggerated 2B4 expression on iNKT cells of HIV-infected treatment-naïve individuals. In sharp contrast to CD4(-)iNKT cells, 2B4 expression was significantly higher on CD4(+) iNKT cell subset. Notably, an increased level of 2B4 on iNKT cells was strongly correlated with parameters associated with HIV disease progression. Further, iNKT cells from ART-naïve individuals were defective in their ability to produce intracellular IFN-γ. Together, our results suggest that the levels ...
International Archives of Allergy and Immunology, 2016
Aim of Study: We used a triplex real-time polymerase chain reaction (PCR) to classify our common ... more Aim of Study: We used a triplex real-time polymerase chain reaction (PCR) to classify our common variable immunodeficiency (CVID) patients into distinct groups according to the amount of their T-cell receptor excision circles (TRECs) and κ-deleting recombination excision circles (KRECs). Materials and Methods: TREC and KREC analysis was performed using a multiplex real-time PCR assay. The T- and B-lymphocyte subsets were measured by flow cytometry. Results: The copy number of TRECs and KRECs was significantly reduced in CVID patients compared to healthy controls. The TREC copy number was inversely correlated with age in both healthy subjects and patients; however, the KREC copy number was inversely correlated with age only in CVID patients. Moreover, no association was seen between TREC/KREC copy number and clinical manifestations such as bronchiectasis, splenomegaly, granulomata, autoimmune cytopenias, organ-specific autoimmunity, enteropathy and lymphoid hyperplasia. Conclusion: T...
Summary Immune senescence as well as disturbed CD8+ T cell differentiation are a hallmark of chro... more Summary Immune senescence as well as disturbed CD8+ T cell differentiation are a hallmark of chronic HIV infection. Here, we investigated to what extent immune senescence is reversible after initiation of anti-retroviral treatment (ART). Peripheral blood mononuclear cells (PBMCs) from a cohort of HIV patients with different disease courses, including untreated viral controllers (n = 10), viral non-controllers (n = 16) and patients on ART (n = 20), were analysed and compared to uninfected controls (n = 25) by flow cytometry on bulk and HIV-specific major histocompatibility complex (MHC) class I tetramer+ CD8+ T cells for expression of the memory markers CCR7 and CD45RO, as well as the senescence marker CD57 and the differentiation and survival marker CD127. Furthermore, a subset of patients was analysed longitudinally before and after initiation of ART. Frequencies of CD57+CD8+ T cells decreased after initiation of ART in central memory (Tcm) but not in effector memory T cell populat...
To function optimally, human blood natural killer (NK) cells need to communicate with other immun... more To function optimally, human blood natural killer (NK) cells need to communicate with other immune cells. Previously, it has been shown that NK cells communicate with 6-sulfo LacNAc dendritic cells (slanDCs), which are able to stimulate NK cells in vitro. In this study, we investigated how slanDCs regulate the level of NK cell activation. The secretion of interleukin (IL)-1β by slanDCs during coculture with NK cells increased as a result of signaling via intercellular adhesion molecule-1 on slanDCs following its interaction with lymphocyte function-associated antigen-1 on NK cells. IL-1β induced the expression of Fas receptor (CD95) on NK cells. The binding of Fas ligand (CD178) to CD95 induced the apoptosis of activated NK cells. Moreover, IL-1β also induced increased cyclooxygenase-2 expression in slanDCs, which in turn enabled the cells to secrete prostaglandin (PG)-E2. Consequently, PGE2 acted as a suppressing agent, tuning down the activation level of NK cells. In summary, IL-1...
HIV-HCV co-infection is associated with accelerated progression to hepatic fibrosis, cirrhosis an... more HIV-HCV co-infection is associated with accelerated progression to hepatic fibrosis, cirrhosis and hepatocellular carcinoma than HCV mono-infection. The contribution of innate immunity during HIV-HCV co-infection has been a relatively under-investigated area. Natural killer (NK) cells are pivotal sentinels of innate immunity against viruses and tumour cells. In this study we evaluated the effect of HIV-HCV co-infection on peripheral blood NK cell subsets with emphasis on the phenotype of CD56(bright) NK cells. Sixty patients were included in the study; HIV mono-infected (n = 12), HCV mono-infected (n = 15), HCV-HIV co-infected (n = 21) and healthy controls (n = 16). PBMCs were isolated and immunophenotyping of NK cells was performed by flowcytometry. We observed an expansion of CD56(bright) NK cell subset in HIV-HCV co-infection as compared to healthy controls and HIV mono-infected group. All the infected groups had an upregulated expression of the activating receptor NKG2D on CD56(...
Cellular and molecular life sciences : CMLS, Jan 5, 2015
Natural killer (NK) cells are innate immune effectors that provide first line of defence against ... more Natural killer (NK) cells are innate immune effectors that provide first line of defence against viruses. Human NK cells are heterogeneous in nature, and their functions rely on a dynamic balance between germ-line-encoded activating and inhibitory receptors. HIV-1 infection results in altered NK cell receptor repertoire and impaired effector functions including the ability to lyse virus-infected cells and secretion of antiviral cytokine IFN-γ. Over the last decade, additional NK cell subset-specific molecules have been identified, leading to emergence of a more complex cellular diversity than previously thought. Herein, we discuss NK cell subset redistribution, altered receptor repertoire and influence of interaction of polymorphic leucocyte antigen (HLA) and killer cell immunoglobulin-like receptors (KIR) on HIV-1 disease progression.
Natural killer (NK) cells are effector and regulatory innate immune cells and play a critical rol... more Natural killer (NK) cells are effector and regulatory innate immune cells and play a critical role in the first line of defense against various viral infections. Although previous reports have indicated the vital contributions of NK cells to HIV-1 immune control, nongenetic NK cell parameters directly associated with slower disease progression have not been defined yet. In a longitudinal, retrospective study of 117 untreated HIV-infected subjects, we show that higher frequencies as well as the absolute numbers of CD8(+) CD3(-) lymphocytes are linked to delayed HIV-1 disease progression. We show that the majority of these cells are well-described blood NK cells. In a subsequent cross-sectional study, we demonstrate a significant loss of CD8(+) NK cells in untreated HIV-infected individuals, which correlated with HIV loads and inversely correlated with CD4(+) T cell counts. CD8(+) NK cells had modestly higher frequencies of CD57-expressing cells than CD8(-) cells, but CD8(+) and CD8(-...
NK cells are pivotal sentinels of the innate immune system and distinct subpopulations in periphe... more NK cells are pivotal sentinels of the innate immune system and distinct subpopulations in peripheral blood have been described. A number of studies addressed HIV-induced alterations of NK cell phenotype and functionality mainly focusing on CD56 dim CD16 + and CD56 2 CD16 + NK cells. However, the impact of HIV-infection on CD56 bright NK cells is less well understood. Here we report a rise of CD56 bright NK cells in HIV-infected individuals, which lack CCR7-expression and strongly correlate with HIV viral load. CCR7 2 CD56 bright NK cells were characterized by increased cytolytic potential, higher activation states and a more differentiated phenotype. These cells thus acquired a number of features of CD56 dim CD16 + NK cells. Furthermore, CD56 bright NK cells from HIV patients exhibited higher degranulation levels compared to uninfected individuals. Thus, chronic HIV-infection is associated with a phenotypic and functional shift of CD56 bright NK cells, which provides a novel aspect of HIV-associated pathogenesis within the NK cell compartment.
Coxiella burnetii (Cb) is a hardy, stealth bacterial pathogen lethal for humans and animals. Its ... more Coxiella burnetii (Cb) is a hardy, stealth bacterial pathogen lethal for humans and animals. Its tremendous resistance to the environment, ease of propagation, and incredibly low infectious dosage make it an attractive organism for biowarfare. Current research on the classification of Coxiella and features influencing its presence in the soil is generally confined to statistical techniques. Machine learning other than traditional approaches can help us better predict epidemiological modeling for this soil-based pathogen of public significance. We developed a two-phase feature-ranking technique for the pathogen on a new soil feature dataset. The feature ranking applies methods such as ReliefF (RLF), OneR (ONR), and correlation (CR) for the first phase and a combination of techniques utilizing weighted scores to determine the final soil attribute ranks in the second phase. Different classification methods such as Support Vector Machine (SVM), Linear Discriminant Analysis (LDA), Logist...
The aim of this study is to analyse the influence of LILRA3 and the genetic leukocyte immunoglobu... more The aim of this study is to analyse the influence of LILRA3 and the genetic leukocyte immunoglobulin-like receptor 3 (LILRA3) deletion on transmission and clinical course of HIV infection. Case and control study. LILRA3 genotypes were determined by PCR. HIV patients were categorized into short-term progressors, normal progressors and long-term nonprogressors according to the clinical course. Functional studies were performed using real-time PCR, intracellular flow cytometry and ELISA. The prevalence of the homozygous LILRA3 deletion was higher in HIV-positive individuals (n = 439) than in controls (n = 651) (P = 0.02). The disease progression was faster in homozygously deleted patients with more short-term progressors than in heterozygous (P = 0.03) and homozygously positive (P = 0.002) individuals. These results have been confirmed in a seroconverter cohort (n = 288). The frequency of the homozygous deletion in the confirmation cohort was higher than in controls (P = 0.04). Combini...
Human natural killer (NK) cells are one major component of lymphocytes that mediate early protect... more Human natural killer (NK) cells are one major component of lymphocytes that mediate early protection against viruses and tumor cells, and play an important role in immune regulatory functions. In this study, we demonstrated that human NK cells could be divided into four subsets, CD56 hi CD16 À , CD56 lo CD16 À , CD56 þ CD16 þ and CD56 À CD16 þ , based on the expression of cell surface CD56 and CD16 molecules. Phenotypic analysis of NK cell subsets indicated that the expression of activation markers, adhesion molecules, memory cell markers, inhibitory and activating receptors, and intracellular proteins (granzyme B and perforin) were heterogeneous. Following interleukin (IL)-2 stimulation, interferon-g was preferentially produced by CD56 þ CD16 À NK cells and this subset showed more proliferative capacity. The cytolytic activity of both CD56 þ CD16 À and CD56 þ/À CD16 þ subsets could be augmented in response to IL-2. The data provided a new definition for NK cell subsets demonstrating their phenotypic and functional diversity and possible stage of NK cell differentiation in peripheral blood.
Cooperative learning is being used throughout the world. It is gaining international popularity. ... more Cooperative learning is being used throughout the world. It is gaining international popularity. On the basis of theory and practice, cooperative learning methods have been used and tested in different cultures and classroom situations. Therefore, a study has been conducted in order to examine the "comparative effectiveness of cooperative learning and traditional approaches of teaching Islamic Studies at Elementary level in Pakistan". Pretest posttest experimental design was used. Students of class six were equally divided into two groups on the basis of teacher made pretest scores. As a treatment cooperative learning method Student-Teams-Achievement-Division (STAD) was provided to experimental group, while control group was taught by using traditional methods of teaching, and kept under traditional classroom situation for the period of 12 weeks. At the end of the treatment, teacher made posttest was administered to measure the achievement scores of students. Significance of difference between the mean scores of both groups was tested by applying t-test at 0.05 level of significance. Findings of the study revealed that both the groups were equal in the beginning of the experiment. The experimental group showed better results on posttest scores, showing the obvious preeminence of cooperative learning method over traditional methods of teaching. High and low achievers of experimental group showed better result as compared to control group, and also helpful in developing creative thinking. Results of the study indicate that cooperative learning is more effective teaching method for Islamic Studies as compared to traditional methods. It revealed that it was more 2 REVIEW OF THE RELATED LITERATURE 9 xvi APPENDICES Appendix Topic
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