The functional integrity of the cytoskeleton of mesenchymal stem cells (MSCs) is essential for it... more The functional integrity of the cytoskeleton of mesenchymal stem cells (MSCs) is essential for its differentiation into multiple cell lineages including adipocytes, chondrocytes, and osteoblasts. Abnormalities in the cytoskeletal proteins such as actin and microtubule can cause disrupted cell signalling and irregular movements of organelles leading to cell death. This study investigated cytoskeletal and nuclear morphological changes of the MSC due to oxidative damage by hydrogen peroxide (H 2 O 2) and the possible prevention of these changes by the antioxidant thymoquinone (TQ). Bone marrow MSCs from Sprague Dawley rats were cultured and treated with different concentrations of H 2 O 2 with or without TQ to observe the potential protective activity. Triple-label fluorescence immunocytochemistry was performed post-treatment to observe the nucleus, actin and microtubules using 4',6-diamidino-2-phenylindole (DAPI), Alexa Fluor 488-labelled phalloidin and Cy3-labelled anti-tubulin antibody, respectively. The normal stem cell cytoskeleton demonstrated intact actin and microtubule structures along with normal appearance of the nucleus. However, oxidative damage by H 2 O 2 caused a severe disruption of the cytoskeletal morphology of the actin and microtubule along with apoptosis and necrosis of the nucleus. Interestingly, both immunocytochemical and Fluorescence-Activated Cell Sorting (FACS) results showed that these morphological changes were prevented by TQ at low concentrations while higher concentrations of TQ were harmful. This study suggested that TQ could save MSCs from oxidative-induced cell death.
ETHNOPHARMACOLOGICAL RELEVANCE The edible plant Opuntia dillenii (Ker Gawl.) Haw. commonly known ... more ETHNOPHARMACOLOGICAL RELEVANCE The edible plant Opuntia dillenii (Ker Gawl.) Haw. commonly known as Nagphana, belongs to the Cactaceae family. It is traditionally used to treat various ailments including inflammation, gastric ulcers, diabetes, hepatitis, asthma, whooping cough and intestinal spasm. AIM OF THE STUDY Despite its traditional use in various countries, detailed toxicological studies of O. dillenii cladode are few. Thus in the current study, toxicity of O. dillenii cladode derived methanol extract, fractions and its α-pyrones: opuntiol and opuntioside have been addressed. METHODS The test agents were assessed using both in vitro and in vivo toxicity assays. MTT on human embryonic kidney cell line (HEK-293), tryphan blue exclusion in rat neutrophils, Cytokinesis-B block micronucleus (CBMN) in human lymphocytes and genomic DNA fragmentation using agarose gel electrophoresis were performed. In acute toxicity test, mice orally received extract (5 g/kg) for 7 days followed by measurements of relative organ weight, biochemical (blood profile, liver and kidney function test) and histological studies (liver and kidney) were carried out. Rat bone marrow micronucleus genotoxicity assay was also conducted. RESULTS O. dillenii derived test agents were non-cytotoxic and had no effect on the integrity of DNA. Methanol extract (5 g/kg) orally administered in mice did not cause any significant change in relative organ weights, biochemical parameters and liver and kidney histology as compared to vehicle control. In parallel, extract did not stimulate micronuclei formation in rat bone marrow polychromatic erythrocytes. CONCLUSION These results led to conclude that edible O. dillenii extract is non-toxic via the oral route and appears to be non-cyto-, hepato-, nephro- or genotoxic, thereby supporting its safe traditional use against various ailments. Therefore, opuntiol and opuntioside may serve as lead compounds in designing new drug(s) derived from edible plants.
Ruzali, Nurul Kabir Nurul Bashar, and Durriyyah Sharifah Hasan Adli (2015). Senile dementia from ... more Ruzali, Nurul Kabir Nurul Bashar, and Durriyyah Sharifah Hasan Adli (2015). Senile dementia from neuroscientific and Islamic
Iron deficiency anemia is one of the most common micronutrient deficient conditions around the gl... more Iron deficiency anemia is one of the most common micronutrient deficient conditions around the globe with various consequences, including the weakened immune system. Quercetin is widely distributed bioflavonoid; it has been debated for its dual roles in iron regulation. Quercetin-iron interaction in the body is a complex mechanism which has not been completely understood. The present study aimed to investigate the effect of quercetin on iron supplementation in iron deficiency anemia and on iNOS expression in splenic macrophages. The rat model of iron deficiency anemia was induced by feeding low iron diet to weanling rats for 20 days. The animals were then administered with ferrous sulfate, quercetin, and their combination for 30 days. Blood parameters, histopathological analysis, iron storage, CD68, iNOS and SLC40 expression in rat spleen were investigated. Our results showed that quercetin regulated iron absorption, despite SLC40 down-expression, indicating possible alternate route of iron transport, and that quercetin modulated iNOS production in splenic macrophages. .
Pasteurella multocida is the main cause of haemorrhagic septicaemia (HS) outbreak in livestock, s... more Pasteurella multocida is the main cause of haemorrhagic septicaemia (HS) outbreak in livestock, such as cattle and buffaloes. Conventional vaccines such as alum-precipitated or oil-adjuvant broth bacterins were injected subcutaneously to provide protection against HS. However, the immunity developed is only for short term and needed to be administered frequently. In our previous study, a short gene fragment from Pasteurella multocida serotype B was obtained via shotgun cloning technique and later was cloned into bacterial expression system. pQE32-ABA392 was found to possess immunogenic activity towards HS when tested in vivo in rat model. In this study, the targeted gene fragment of ABA392 was sub-cloned into a DNA expression vector pVAX1 and named as pVAX1-ABA392. The new recombinant vaccine was stable and expressed on mammalian cell lines. Serum sample collected from a group of vaccinated rats for ELISA test shows that the antibody in immunized rats was present at high titer and can be tested as a vaccine candidate with challenge in further studies. This successful recombinant vaccine is immunogenic and potentially could be used as vaccine in future against HS.
Aims: The aim of the current study was to identify active compound(s) responsible for the antipro... more Aims: The aim of the current study was to identify active compound(s) responsible for the antiproliferative effects of O. basilicum and explore their underlying mechanism/s. Background: Plants have been the source of medicines for the treatment of various diseases since ancient times. Ocimum basilicum (Sweet Basil, Bobai Tulsi) has been used in the folk medicine for the treatment of human liver, spleen and stomach cancers. Objective: To emphasize the importance of O. basilicum as a potential novel non-toxic alternative to the conventional anticancer therapy. Method: O. basilicum (aerial parts) methanolic extract and fractions were screened against HT-144, MCF-7, NCI-H460 and SF-268 human cancer cell lines using sulforhodamine B assay. The more active petroleum ether insoluble (PEI) fraction was fractionated into six sub-fractions (OB-1 to OB-6). Four pure compounds (3-O-methyl ursolic acid, oleanolic acid, 3-epi-ursolic acid and ursolic acid) were isolated from the more potent sub-f...
Significance and Impact of the Study: New approach of combating haemorrhagic septicaemia disease ... more Significance and Impact of the Study: New approach of combating haemorrhagic septicaemia disease among bovines by recombinant DNA vaccine is crucial to overcome the loss of edible products from the infected bovines. DNA vaccine can potentially serve as a better immunogen which would elicit both cellular and humoral immunity, and it is also stable for its molecular reproduction. This research report demonstrates an effective yet simple way of administering the DNA vaccine via the intranasal route in rats, to provoke the mucosal immunity through the development of immunoglobulins IgA, IgG and bronchus-associated lymphoid tissue which guard as the first-line defence at the host's mucosal lining.
Haemorrhagic septicaemia (HS) is a well-known high fatality septicaemic disease happening among b... more Haemorrhagic septicaemia (HS) is a well-known high fatality septicaemic disease happening among bovines. The disease is caused by the Pasteurella multocida serotype B:2 bacteria. P. multocida B:2 has high mortality and morbidity rates and is spread through the intranasal and oral routes in bovines. In this study, our aim was to investigate the efficacy of the recombinant protein vaccine, ABA392/pET30a via intranasal inoculation by targeting the mucosal immunity. The constructed recombinant protein vaccine ABA392/pET30a was subjected to an animal study using Sprague Dawley rats. The study was divided into two parts: active and passive immunization studies. Both studies were carried out through the determination of immunogenicity (using Total White Blood Cell (TWBC) Count with Indirect ELISA) and histopathogenicity, analyzing (Bronchus Associated Lymphoid Tissue (BALT) formation) in lungs. As a result, the IgA and IgG development of both tested groups: group 1 (50μg/mL protein vaccine) and group 2 (100μg/mL protein vaccine) showed equivalent with the positive control group 4 (formalin-killed P. multocida B:2). However, there was a significant difference when compared with the negative control group 3 (normal saline). These results demonstrate that both the protein vaccine at the concentration 50μg/mL and 100μg/mL have the same efficacy as the commercially available positive control vaccine. From the studies, higher concentration of protein vaccine at 100μg/mL showed higher development of both IgA and IgG compared to 50μg/mL protein vaccine. Higher and rapid development of IgA compared to IgG showed that mucosal immunity has been induced through the intranasal administration of the protein vaccine. In addition, leucocytosis was observed at each dose of vaccination showed that the protein vaccine is capable to induce the immune responses of the host. Histopathogenicity studies of the vaccinated groups showed more BALT formation and no severe lesions after challenge compared to the negative control group. Besides, no inflammatory onsite or anaphylactic responses were observed after the intranasal inoculation which proved to be safer and provided longer lasting immunity. Therefore, recombinant protein vaccine ABA392/pET30a could be a potential candidate for intranasal administration which can provoke mucosal immunity against HS disease.
For a long time, hematoxylin and eosin (H&E) staining has been widely used for the diagnosis of m... more For a long time, hematoxylin and eosin (H&E) staining has been widely used for the diagnosis of many diseases by assessing the histopathological changes in tissue sections. Eosin, one of the staining dyes in the H&E stain, is fluorescent in the fluorescein isothiocyanate (FITC) and Texas Red (TxR) fluorescent channels. In this study, the use of eosin fluorescence to visualize and quantify histopathological changes was elucidated in a glycerolinduced model of acute kidney injury (AKI) in mice. AKI was induced in BALB/c mice by intramuscularly injecting 10 mL/kg of 50% glycerol after 24 h of water deprivation and, after another 24 h, the kidneys were dissected out. After H&E staining, kidney tissue sections were examined under a fluorescent microscope using a doublechannel filter cube (FITC and TxR). The H&E fluorescence pattern of normal kidney structures was characterized as the fluorescence patterns varied between different parts of the kidney and could be used for the structural identification of different parts of the kidney. Interestingly, in the injured kidney sections, a striking increase in the red fluorescence was noticed only in the damaged areas. In the damaged kidney sections, casts deposited inside tubules showed bright yellow fluorescence, whereas fibers showed strong green fluorescence. In vitro experiments were also performed in order to understand possible mechanisms underlying this phenomenon. It was concluded that eosin fluorescence can be used to quantify the degree of damage to kidney tissue which, in turn, may help in assessing the kidney-protective effects of unknown compounds and plant extracts.
Introduction: Rap1, a member of Ras superfamily of small GTP-binding proteins, is involved in car... more Introduction: Rap1, a member of Ras superfamily of small GTP-binding proteins, is involved in cardiovascular biology in numerous ways. It is an evolutionary conserved regulator of adhesion, polarity, differentiation and growth. Aims: Our aim was to analyze Rap1 activated rat bone marrow mesenchymal stem cells (MSCs) for their potential role in adhesion and cardiac differentiation. Methods: Myocardial infarction (MI) was produced in Sprague Dawley (SD) rats through occlusion of the left anterior descending coronary artery. MSCs were treated with 8-pCPT-2'-O-Me-cAMP (CPT) to activate Rap1. Normal (untreated) and CPT-treated MSCs were transplanted through intra-myocardial injection in respective groups. Cardiac function was assessed by echocardiography at 2 and 4 weeks after cell transplantation. Histological analysis was performed to observe changes at tissue level. Results: Homing of CPT-treated MSCs was significantly (***P < 0.001) higher as compared to normal MSCs in the infarcted hearts. This may be due to increase in the gene expression of some of the cell adhesion molecules as evident by qRT-PCR analysis. Significant (***P < 0.001) improvement in the restoration of heart function in terms of left ventricular diastolic and systolic internal diameters (LVIDd, LVIDs), % ejection fraction, % fraction shortening, and end systolic and diastolic volumes were observed in CPT-treated MSCs as compared to the MI model. Histological analyses showed significant (***P < 0.001) Accepted Article This article is protected by copyright. All rights reserved. reduction in scar formation in the CPT-treated group. Differentiation of treated MSCs into functional cardiomyocytes was evident through immunohistochemical staining. LV wall thickness was also preserved significantly (***P < 0.001). Blood vessel formation was more pronounced in CPT-treated group though both cell therapy groups showed significant increase as compared to MI model. Conclusion: Our findings showed that pharmacological activation of EPAC-Rap1 improves cardiac function through better survival, adhesion, and differentiation of transplanted cells. Transplantation of these MSCs in the infarct area restored functional myocardium.
The present study was undertaken to explore the possible anti-diabetic mechanism(s) of Emblica of... more The present study was undertaken to explore the possible anti-diabetic mechanism(s) of Emblica officinalis (EO) and its active constituent, ellagic acid (EA), in vitro and in vivo. Neonatal streptozotocin-induced non-obese type 2 diabetic rats were treated with a methanolic extract of EO (250 or 500 mg/kg) for 28 days, and blood glucose, serum insulin, and plasma antioxidant status were measured. Insulin and glucagon immunostaining and morphometry were performed in pancreatic section, and liver TBARS and GSH levels were measured. Additionally, EA was tested for glucose-stimulated insulin secretion and glucose tolerance test. Treatment with EO extract resulted in a significant decrease in the fasting blood glucose in a dose- and time-dependent manner in the diabetic rats. It significantly increased serum insulin in the diabetic rats in a dose-dependent manner. Insulin-to-glucose ratio was also increased by EO treatment. Immunostaining of pancreas showed that EO250 increased β-cell size, but EO500 increased β-cells number in diabetic rats. EO significantly increased plasma total antioxidants and liver GSH and decreased liver TBARS. EA stimulated glucose-stimulated insulin secretion from isolated islets and decreased glucose intolerance in diabetic rats. Ellagic acid in EO exerts anti-diabetic activity through the action on β-cells of pancreas that stimulates insulin secretion and decreases glucose intolerance.
The functional integrity of the cytoskeleton of mesenchymal stem cells (MSCs) is essential for it... more The functional integrity of the cytoskeleton of mesenchymal stem cells (MSCs) is essential for its differentiation into multiple cell lineages including adipocytes, chondrocytes, and osteoblasts. Abnormalities in the cytoskeletal proteins such as actin and microtubule can cause disrupted cell signalling and irregular movements of organelles leading to cell death. This study investigated cytoskeletal and nuclear morphological changes of the MSC due to oxidative damage by hydrogen peroxide (H 2 O 2) and the possible prevention of these changes by the antioxidant thymoquinone (TQ). Bone marrow MSCs from Sprague Dawley rats were cultured and treated with different concentrations of H 2 O 2 with or without TQ to observe the potential protective activity. Triple-label fluorescence immunocytochemistry was performed post-treatment to observe the nucleus, actin and microtubules using 4',6-diamidino-2-phenylindole (DAPI), Alexa Fluor 488-labelled phalloidin and Cy3-labelled anti-tubulin antibody, respectively. The normal stem cell cytoskeleton demonstrated intact actin and microtubule structures along with normal appearance of the nucleus. However, oxidative damage by H 2 O 2 caused a severe disruption of the cytoskeletal morphology of the actin and microtubule along with apoptosis and necrosis of the nucleus. Interestingly, both immunocytochemical and Fluorescence-Activated Cell Sorting (FACS) results showed that these morphological changes were prevented by TQ at low concentrations while higher concentrations of TQ were harmful. This study suggested that TQ could save MSCs from oxidative-induced cell death.
ETHNOPHARMACOLOGICAL RELEVANCE The edible plant Opuntia dillenii (Ker Gawl.) Haw. commonly known ... more ETHNOPHARMACOLOGICAL RELEVANCE The edible plant Opuntia dillenii (Ker Gawl.) Haw. commonly known as Nagphana, belongs to the Cactaceae family. It is traditionally used to treat various ailments including inflammation, gastric ulcers, diabetes, hepatitis, asthma, whooping cough and intestinal spasm. AIM OF THE STUDY Despite its traditional use in various countries, detailed toxicological studies of O. dillenii cladode are few. Thus in the current study, toxicity of O. dillenii cladode derived methanol extract, fractions and its α-pyrones: opuntiol and opuntioside have been addressed. METHODS The test agents were assessed using both in vitro and in vivo toxicity assays. MTT on human embryonic kidney cell line (HEK-293), tryphan blue exclusion in rat neutrophils, Cytokinesis-B block micronucleus (CBMN) in human lymphocytes and genomic DNA fragmentation using agarose gel electrophoresis were performed. In acute toxicity test, mice orally received extract (5 g/kg) for 7 days followed by measurements of relative organ weight, biochemical (blood profile, liver and kidney function test) and histological studies (liver and kidney) were carried out. Rat bone marrow micronucleus genotoxicity assay was also conducted. RESULTS O. dillenii derived test agents were non-cytotoxic and had no effect on the integrity of DNA. Methanol extract (5 g/kg) orally administered in mice did not cause any significant change in relative organ weights, biochemical parameters and liver and kidney histology as compared to vehicle control. In parallel, extract did not stimulate micronuclei formation in rat bone marrow polychromatic erythrocytes. CONCLUSION These results led to conclude that edible O. dillenii extract is non-toxic via the oral route and appears to be non-cyto-, hepato-, nephro- or genotoxic, thereby supporting its safe traditional use against various ailments. Therefore, opuntiol and opuntioside may serve as lead compounds in designing new drug(s) derived from edible plants.
Ruzali, Nurul Kabir Nurul Bashar, and Durriyyah Sharifah Hasan Adli (2015). Senile dementia from ... more Ruzali, Nurul Kabir Nurul Bashar, and Durriyyah Sharifah Hasan Adli (2015). Senile dementia from neuroscientific and Islamic
Iron deficiency anemia is one of the most common micronutrient deficient conditions around the gl... more Iron deficiency anemia is one of the most common micronutrient deficient conditions around the globe with various consequences, including the weakened immune system. Quercetin is widely distributed bioflavonoid; it has been debated for its dual roles in iron regulation. Quercetin-iron interaction in the body is a complex mechanism which has not been completely understood. The present study aimed to investigate the effect of quercetin on iron supplementation in iron deficiency anemia and on iNOS expression in splenic macrophages. The rat model of iron deficiency anemia was induced by feeding low iron diet to weanling rats for 20 days. The animals were then administered with ferrous sulfate, quercetin, and their combination for 30 days. Blood parameters, histopathological analysis, iron storage, CD68, iNOS and SLC40 expression in rat spleen were investigated. Our results showed that quercetin regulated iron absorption, despite SLC40 down-expression, indicating possible alternate route of iron transport, and that quercetin modulated iNOS production in splenic macrophages. .
Pasteurella multocida is the main cause of haemorrhagic septicaemia (HS) outbreak in livestock, s... more Pasteurella multocida is the main cause of haemorrhagic septicaemia (HS) outbreak in livestock, such as cattle and buffaloes. Conventional vaccines such as alum-precipitated or oil-adjuvant broth bacterins were injected subcutaneously to provide protection against HS. However, the immunity developed is only for short term and needed to be administered frequently. In our previous study, a short gene fragment from Pasteurella multocida serotype B was obtained via shotgun cloning technique and later was cloned into bacterial expression system. pQE32-ABA392 was found to possess immunogenic activity towards HS when tested in vivo in rat model. In this study, the targeted gene fragment of ABA392 was sub-cloned into a DNA expression vector pVAX1 and named as pVAX1-ABA392. The new recombinant vaccine was stable and expressed on mammalian cell lines. Serum sample collected from a group of vaccinated rats for ELISA test shows that the antibody in immunized rats was present at high titer and can be tested as a vaccine candidate with challenge in further studies. This successful recombinant vaccine is immunogenic and potentially could be used as vaccine in future against HS.
Aims: The aim of the current study was to identify active compound(s) responsible for the antipro... more Aims: The aim of the current study was to identify active compound(s) responsible for the antiproliferative effects of O. basilicum and explore their underlying mechanism/s. Background: Plants have been the source of medicines for the treatment of various diseases since ancient times. Ocimum basilicum (Sweet Basil, Bobai Tulsi) has been used in the folk medicine for the treatment of human liver, spleen and stomach cancers. Objective: To emphasize the importance of O. basilicum as a potential novel non-toxic alternative to the conventional anticancer therapy. Method: O. basilicum (aerial parts) methanolic extract and fractions were screened against HT-144, MCF-7, NCI-H460 and SF-268 human cancer cell lines using sulforhodamine B assay. The more active petroleum ether insoluble (PEI) fraction was fractionated into six sub-fractions (OB-1 to OB-6). Four pure compounds (3-O-methyl ursolic acid, oleanolic acid, 3-epi-ursolic acid and ursolic acid) were isolated from the more potent sub-f...
Significance and Impact of the Study: New approach of combating haemorrhagic septicaemia disease ... more Significance and Impact of the Study: New approach of combating haemorrhagic septicaemia disease among bovines by recombinant DNA vaccine is crucial to overcome the loss of edible products from the infected bovines. DNA vaccine can potentially serve as a better immunogen which would elicit both cellular and humoral immunity, and it is also stable for its molecular reproduction. This research report demonstrates an effective yet simple way of administering the DNA vaccine via the intranasal route in rats, to provoke the mucosal immunity through the development of immunoglobulins IgA, IgG and bronchus-associated lymphoid tissue which guard as the first-line defence at the host's mucosal lining.
Haemorrhagic septicaemia (HS) is a well-known high fatality septicaemic disease happening among b... more Haemorrhagic septicaemia (HS) is a well-known high fatality septicaemic disease happening among bovines. The disease is caused by the Pasteurella multocida serotype B:2 bacteria. P. multocida B:2 has high mortality and morbidity rates and is spread through the intranasal and oral routes in bovines. In this study, our aim was to investigate the efficacy of the recombinant protein vaccine, ABA392/pET30a via intranasal inoculation by targeting the mucosal immunity. The constructed recombinant protein vaccine ABA392/pET30a was subjected to an animal study using Sprague Dawley rats. The study was divided into two parts: active and passive immunization studies. Both studies were carried out through the determination of immunogenicity (using Total White Blood Cell (TWBC) Count with Indirect ELISA) and histopathogenicity, analyzing (Bronchus Associated Lymphoid Tissue (BALT) formation) in lungs. As a result, the IgA and IgG development of both tested groups: group 1 (50μg/mL protein vaccine) and group 2 (100μg/mL protein vaccine) showed equivalent with the positive control group 4 (formalin-killed P. multocida B:2). However, there was a significant difference when compared with the negative control group 3 (normal saline). These results demonstrate that both the protein vaccine at the concentration 50μg/mL and 100μg/mL have the same efficacy as the commercially available positive control vaccine. From the studies, higher concentration of protein vaccine at 100μg/mL showed higher development of both IgA and IgG compared to 50μg/mL protein vaccine. Higher and rapid development of IgA compared to IgG showed that mucosal immunity has been induced through the intranasal administration of the protein vaccine. In addition, leucocytosis was observed at each dose of vaccination showed that the protein vaccine is capable to induce the immune responses of the host. Histopathogenicity studies of the vaccinated groups showed more BALT formation and no severe lesions after challenge compared to the negative control group. Besides, no inflammatory onsite or anaphylactic responses were observed after the intranasal inoculation which proved to be safer and provided longer lasting immunity. Therefore, recombinant protein vaccine ABA392/pET30a could be a potential candidate for intranasal administration which can provoke mucosal immunity against HS disease.
For a long time, hematoxylin and eosin (H&E) staining has been widely used for the diagnosis of m... more For a long time, hematoxylin and eosin (H&E) staining has been widely used for the diagnosis of many diseases by assessing the histopathological changes in tissue sections. Eosin, one of the staining dyes in the H&E stain, is fluorescent in the fluorescein isothiocyanate (FITC) and Texas Red (TxR) fluorescent channels. In this study, the use of eosin fluorescence to visualize and quantify histopathological changes was elucidated in a glycerolinduced model of acute kidney injury (AKI) in mice. AKI was induced in BALB/c mice by intramuscularly injecting 10 mL/kg of 50% glycerol after 24 h of water deprivation and, after another 24 h, the kidneys were dissected out. After H&E staining, kidney tissue sections were examined under a fluorescent microscope using a doublechannel filter cube (FITC and TxR). The H&E fluorescence pattern of normal kidney structures was characterized as the fluorescence patterns varied between different parts of the kidney and could be used for the structural identification of different parts of the kidney. Interestingly, in the injured kidney sections, a striking increase in the red fluorescence was noticed only in the damaged areas. In the damaged kidney sections, casts deposited inside tubules showed bright yellow fluorescence, whereas fibers showed strong green fluorescence. In vitro experiments were also performed in order to understand possible mechanisms underlying this phenomenon. It was concluded that eosin fluorescence can be used to quantify the degree of damage to kidney tissue which, in turn, may help in assessing the kidney-protective effects of unknown compounds and plant extracts.
Introduction: Rap1, a member of Ras superfamily of small GTP-binding proteins, is involved in car... more Introduction: Rap1, a member of Ras superfamily of small GTP-binding proteins, is involved in cardiovascular biology in numerous ways. It is an evolutionary conserved regulator of adhesion, polarity, differentiation and growth. Aims: Our aim was to analyze Rap1 activated rat bone marrow mesenchymal stem cells (MSCs) for their potential role in adhesion and cardiac differentiation. Methods: Myocardial infarction (MI) was produced in Sprague Dawley (SD) rats through occlusion of the left anterior descending coronary artery. MSCs were treated with 8-pCPT-2'-O-Me-cAMP (CPT) to activate Rap1. Normal (untreated) and CPT-treated MSCs were transplanted through intra-myocardial injection in respective groups. Cardiac function was assessed by echocardiography at 2 and 4 weeks after cell transplantation. Histological analysis was performed to observe changes at tissue level. Results: Homing of CPT-treated MSCs was significantly (***P < 0.001) higher as compared to normal MSCs in the infarcted hearts. This may be due to increase in the gene expression of some of the cell adhesion molecules as evident by qRT-PCR analysis. Significant (***P < 0.001) improvement in the restoration of heart function in terms of left ventricular diastolic and systolic internal diameters (LVIDd, LVIDs), % ejection fraction, % fraction shortening, and end systolic and diastolic volumes were observed in CPT-treated MSCs as compared to the MI model. Histological analyses showed significant (***P < 0.001) Accepted Article This article is protected by copyright. All rights reserved. reduction in scar formation in the CPT-treated group. Differentiation of treated MSCs into functional cardiomyocytes was evident through immunohistochemical staining. LV wall thickness was also preserved significantly (***P < 0.001). Blood vessel formation was more pronounced in CPT-treated group though both cell therapy groups showed significant increase as compared to MI model. Conclusion: Our findings showed that pharmacological activation of EPAC-Rap1 improves cardiac function through better survival, adhesion, and differentiation of transplanted cells. Transplantation of these MSCs in the infarct area restored functional myocardium.
The present study was undertaken to explore the possible anti-diabetic mechanism(s) of Emblica of... more The present study was undertaken to explore the possible anti-diabetic mechanism(s) of Emblica officinalis (EO) and its active constituent, ellagic acid (EA), in vitro and in vivo. Neonatal streptozotocin-induced non-obese type 2 diabetic rats were treated with a methanolic extract of EO (250 or 500 mg/kg) for 28 days, and blood glucose, serum insulin, and plasma antioxidant status were measured. Insulin and glucagon immunostaining and morphometry were performed in pancreatic section, and liver TBARS and GSH levels were measured. Additionally, EA was tested for glucose-stimulated insulin secretion and glucose tolerance test. Treatment with EO extract resulted in a significant decrease in the fasting blood glucose in a dose- and time-dependent manner in the diabetic rats. It significantly increased serum insulin in the diabetic rats in a dose-dependent manner. Insulin-to-glucose ratio was also increased by EO treatment. Immunostaining of pancreas showed that EO250 increased β-cell size, but EO500 increased β-cells number in diabetic rats. EO significantly increased plasma total antioxidants and liver GSH and decreased liver TBARS. EA stimulated glucose-stimulated insulin secretion from isolated islets and decreased glucose intolerance in diabetic rats. Ellagic acid in EO exerts anti-diabetic activity through the action on β-cells of pancreas that stimulates insulin secretion and decreases glucose intolerance.
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