Chorein, a protein supporting activation of phosphoinositide 3 kinase (PI3K), participates in the... more Chorein, a protein supporting activation of phosphoinositide 3 kinase (PI3K), participates in the regulation of actin polymerization and cell survival. A loss of function mutation of the chorein encoding gene VPS13A (vacuolar protein sorting-associated protein 13A) leads to chorea-acanthocytosis (ChAc), a neurodegenerative disorder with simultaneous erythrocyte akanthocytosis. In blood platelets chorein deficiency has been shown to compromise expression of vesicle-associated membrane protein 8 (VAMP8) and thus degranulation. The present study explored whether chorein is similarly involved in VAMP8 expression and dopamine release of pheochromocytoma (PC12) cells. Chorein was down-regulated by silencing in PC12 cells. Transmission electron microscopy was employed to quantify the number of vesicles, RT-PCR to determine transcript levels, Western blotting to quantify protein expression and ELISA to determine dopamine release. Chorein silencing significantly reduced the number of vesicles, VAMP8 transcript levels and VAMP8 protein abundance. Increase of extracellular K+ from 5 mM to 40 mM resulted in marked stimulation of dopamine release, an effect significantly blunted by chorein silencing. Chorein deficiency down-regulates VAMP8 expression, vesicle numbers and dopamine release in pheochromocytoma cells.
Mutation Research/Genetic Toxicology and Environmental Mutagenesis, Jun 15, 2012
The mutagenic potential of titanium dioxide nanoparticles (TiO2-NPs) of an average size 30.6 nm w... more The mutagenic potential of titanium dioxide nanoparticles (TiO2-NPs) of an average size 30.6 nm was investigated using β-galactosidase (lacZ) gene complementation in plasmid pUC19/lacZ− Escherichia coli DH5α system. Plasmid pUC19 was treated with varying concentrations of TiO2-NPs and allowed to transfect the CaCl2-induced competent DH5α cells. The data revealed loss in transformation efficiency of TiO2-NPs treated plasmids as compared to untreated plasmid DNA in DH5α host cells. Induction of multiple mutations in ...
Annals of Hepatology Official Journal of the Mexican Association of Hepatology, Mar 1, 2013
This study aims to investigate whether the SNPs of CXCR1 gene, could predict the likelihood of vi... more This study aims to investigate whether the SNPs of CXCR1 gene, could predict the likelihood of viral persistence and/or disease progression. We investigated the association of two different SNPs (rs2234671, and rs142978743) in 598 normal healthy controls and 662 HBV patients from a Saudi ethnic population. The HBV patients were categorized into inactive carriers (n = 428), active carriers (n = 162), cirrhosis (n = 54) and Cirrhosis-HCC (n = 18) sub-groups. Genetic variants in CXCR1 were determined by polymerase chain reaction (PCR)-based DNA direct sequencing. The frequency of the risk allele 'C' for the SNP, rs2234671 was found to be insignificant when the patient group was compared to the uninfected control group, however, a significant distribution of the allele 'C' of rs2234671 was observed among active HBV carriers + cirrhosis + cirrhosis - HCC vs. inactive HBV carriers with an OR = 1.631 (95% C.I. 1.016-2.616) and p = 0.032. However, no significant association was observed for rs142978743 when the distribution of risk allele was analyzed among the different patient groups (i.e. inactive carriers, active carriers, cirrhosis and HCC). Furthermore, the most common haplotype, Haplo-1 (AG), was found to have an insignificant frequency distribution between HBV cases and controls, while the same haplotype was found to be significantly distributed when active carriers + cirrhosis + cirrhosis - HCC patients were compared to inactive HBV carriers with a frequency of 0.938 and p = 0.0315. Haplo-2 (AC) was also found to be significantly associated with a frequency of 0.058 and p = 0.0163. The CXCR1 polymorphism, rs2234671 was found to be associated with chronic HBV infection and may play a role in disease activity.
Sodium potassium pump (Na+/K+ ATPase) is a validated pharmacological target for the treatment of ... more Sodium potassium pump (Na+/K+ ATPase) is a validated pharmacological target for the treatment of various cardiac conditions. Recent published data with Na+/K+ ATPase inhibitors suggest a potent anti-cancer action of these agents in multiple indications. In the present study, we focus on istaroxime, a Na+/K+ ATPase inhibitor that has shown favorable safety and efficacy properties in cardiac phase II clinical trials. Our experiments in 22 cancer cell lines and in prostate tumors in vivo proved the strong anti-cancer action of this compound. Istaroxime induced apoptosis, affected the key proliferative and apoptotic mediators c-Myc and caspase-3 and modified actin cystoskeleton dynamics and RhoA activity in prostate cancer cells. Interestingly, istaroxime was capable of binding to mAR, a membrane receptor mediating rapid, non-genomic actions of steroids in prostate and other cells. These results support a multi-level action of Na+/K+ ATPase inhibitors in cancer cells and collectively validate istaroxime as a strong re-purposing candidate for further cancer drug development.
Chromium oxide nanoparticles (Cr2O3NPs) are widely used in polymers and paints. In the present st... more Chromium oxide nanoparticles (Cr2O3NPs) are widely used in polymers and paints. In the present study, we aimed to determine the toxicity of Cr2O3NPs in murine fibrosarcoma (L929) cells. The cytotoxicity of Cr2O3NPs was measured by MTT and neutral red uptake assays; Cr2O3NPs had significant cytotoxic effects on L929 cells. Enhancement of intracellular reactive oxygen species was observed in L929 cells after exposure to Cr2O3NPs. Cr2O3NPs produced caspase-3, indicating that exposure to Cr2O3NPs induced apoptosis. After exposure to Cr2O3NPs, the cellular glutathione level decreased and lipid peroxidation, superoxide dismutase, and catalase increased in a dose- and time-dependent manner. By using single-cell gel tests, we also observed increased DNA damage in a Cr2O3NP exposure-duration- and dose-dependent fashion. Cell toxicity and DNA damage may be useful biomarkers for determining the safety of Cr2O3NPs in human and animal health.
FGF23 regulates renal phosphate and vitamin D metabolism. Loss of FGF23 results in massive calcif... more FGF23 regulates renal phosphate and vitamin D metabolism. Loss of FGF23 results in massive calcification and rapid aging. FGF23 production is stimulated by 1,25(OH)2 D3 and NFκB signaling. Here, we report that treatment of UMR106 osteoblast-like cells with 1,25(OH)2 D3 , inducing Fgf23 transcription, resulted in actin polymerization which was blocked by NFκB inhibitor wogonin. Interestingly, 1,25(OH)2 D3 -induced Fgf23 gene transcription was abolished by the actin microfilament-disrupting agent cytochalasin B, as well as by the inhibition of actin-regulating Rac1/PAK1 signaling. Our results provide strong evidence that actin redistribution regulated by the Rac1/PAK1 pathway participates in 1,25(OH)2 D3 -induced Fgf23 gene transcription. This article is protected by copyright. All rights reserved.
Barium oxide nanoparticles (BaONPs) are an important industrial compound and are widely used in p... more Barium oxide nanoparticles (BaONPs) are an important industrial compound and are widely used in polymers and paints. In this study, apoptotic and genotoxic effects of BaONPs in mouse embryonic fibroblast (L929) cells were determined by using single-cell gel test. In vitro cytotoxicity assays were performed to assess BaONPs' toxicity in L929 cells. Mild cytotoxicity was observed in L929 cells due to BaONPs. BaONPs increased lipid peroxidation, catalase, and superoxide dismutase levels and lowered glutathione levels in L929 cells. This was accompanied by concomitant generation of reactive oxygen species and activation of caspase-3 in BaONPs-treated L929 cells. On the other hand, when we exposed L929 cells to BaONPs for 24 and 48 hours (comet assay), there was a duration- and dose-dependent increase in DNA impairment detected in the single-cell gel test. Thus, BaONPs exhibit genotoxic and apoptotic effects in L929 cells, most likely due to initiation of oxidative damage.
Quercetol is a polyphenolic molecule present in vegetables and fruits, and is beneficial to human... more Quercetol is a polyphenolic molecule present in vegetables and fruits, and is beneficial to human and animal health. The current work aimed to test cytotoxic and apoptotic effects of quercetol on HepG2 cells. Quercetol was isolated from Ocimum sanctum and characterized by gas chromatography-tandom mass spectrometry (GC-MS/MS), nuclear magnetic resonance spectroscopy, and Fourier transform infrared spectroscopy. Quercetol (50-600 μg/mL) was examined for cytotoxic activity by tetrazolium salt and neutral red uptake tests and comet assay for genotoxicity, using HepG2 cells, over 24 hours. Data from 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide and neutral red uptake tests demonstrated quercetol-induced cytotoxicity in HepG2 cells in a concentration-dependent manner. With 4',6-diamidino-2-phenylindole staining, a significant induction of chromosomal condensation was observed at 300 μg/mL of quercetol. DNA fragmentation analysis showed that quercetol produced cell death in HepG2 cells in a concentration-dependent manner. Thus, our study suggests that an environmentally relevant concentration of quercetol, which was a chemically standardized extract from O. sanctum, induced cell death and DNA damage in HepG2 cells.
This study compares the diagnostic utility of complexed prostate-specific antigen (cPSA), total P... more This study compares the diagnostic utility of complexed prostate-specific antigen (cPSA), total PSA (tPSA) and their ratios with free PSA (fPSA) for benign prostatic hyperplasia (BPH) and prostate cancer. This is though to be the first study to evaluate cPSA in the ethnic population of Saudi Arabia. Serum samples were collected from 54 patients (aged over 50) and assayed for tPSA, cPSA and fPSA. Thirty-five patients were histologically and clinically proven to have BPH and 19 patients were proven to have cancer. Sensitivity, specificity and ROC curves were calculated. With a cPSA cut-off of 4 ng/mL the sensitivity was 79%, the specificity was 34%, and the positive and negative predictive values (PPV and NPV) were 39% and 75%, respectively. At the same cut-off for tPSA, the sensitivity was 84%, the specificity was 29%, and the PPV and NPV were 39 and 77%, respectively. The sensitivity for both tests was lower at a cut-off of 20 ng/mL but the specificity increased to 77% for cPSA and ...
This study was conducted to investigate the ultrastructural changes in the liver and kidney of mi... more This study was conducted to investigate the ultrastructural changes in the liver and kidney of mice induced by a single dose of Cadmium chloride. A group of mice (25) were injected with a single intraperitoneal dose of Cadmium chloride (2 mg/ kg) body weight. After 4 weeks, all treated and untreated animals were killed, the live and kidney were removed and processed for transmission electron microscopy. Ultrastructure of liver tissues showed increase in the number of lysosomal structure and oil droplets. Also changes in the mitochondrial structures and enlarge in Kupffer cells were observed. Ultrastructure of kidney tubule cells showed cellular necrosis, disappearing of microvilli and changes in the mitochondria and nucleus. Basing upon the results of electron microscopy investigation, it was possible to conclude that a single PI dose of cadmium chloride (2mg/kg) can induce ultrastructural changes in both liver and kidney tissues
Journal of Environmental Pathology, Toxicology and Oncology, 2015
This study aims to investigate whether asiaticoside, a triterpene glycoside, can afford protectio... more This study aims to investigate whether asiaticoside, a triterpene glycoside, can afford protection to DNA from alterations induced by gamma radiation under in vitro, ex vivo, and in vivo conditions. In vitro studies were done on plasmid pBR322 DNA, ex vivo studies were done on cellular DNA of human peripheral blood leukocytes, and in vivo investigations were conducted on cellular DNA of spleen and bone marrow cells of mice exposed to whole-body gamma radiation. The supercoiled form of the plasmid pBR322 DNA upon exposure to the radiation was converted into relaxed open circular form due to induction of strand breaks. Presence of asiaticoside along with the DNA during irradiation prevented the relaxation of the supercoiled form to the open circular form. When human peripheral blood leukocytes were exposed to gamma radiation, the cellular DNA suffered strand breaks as evidenced by the increased comet parameters in an alkaline comet assay. Asiaticoside, when present along with blood during irradiation ex vivo, prevented the strand breaks and the comet parameters were closer to that of the controls. Whole-body exposure of mice to gamma radiation resulted in a significant increase in comet parameters of DNA of bone marrow and spleen cells of mice as a result of radiation-induced strand breaks in DNA. Administration of asiaticoside prior to whole-body radiation exposure of the mice prevented this increase in radiation-induced increase in comet parameters, which could be the result of protection to DNA under in vivo conditions of radiation exposure. Thus, it can be concluded from the results that asiaticoside can offer protection to DNA from radiation-induced alterations under in vitro, ex vivo, and in vivo conditions.
Stigmasterol (99.9% pure) was isolated from Azadirachta indica and its chemopreventive effect on ... more Stigmasterol (99.9% pure) was isolated from Azadirachta indica and its chemopreventive effect on 7,12-dimethylbenz[a]anthracene (DMBA)-induced skin cancer was investigated in Swiss albino mice. Skin tumors were induced by topical application of DMBA and promoted by croton oil. To assess the chemopreventive potential of stigmasterol, it was orally administered at a concentration of 200 mg/kg and 400 mg/kg three times weekly for 16 weeks. Reduction in tumor size and cumulative number of papillomas were seen as a result of treatment with stigmasterol. The average latency period was significantly increased as compared with the carcinogen-treated control. Stigmasterol induced a significant decrease in the activity of serum enzymes, such as aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, and bilirubin as compared with the control. Stigmasterol significantly increased glutathione, superoxide dismutase, and catalase as compared with the control. Elevated levels of lipid peroxide and DNA damage in the control group were significantly inhibited by administration of stigmasterol. From the present study, it can be inferred that stigmasterol has chemopreventive activity in an experimental model of cancer. This chemopreventive activity may be linked to the oxidative stress of stigmasterol. The antigenotoxic properties of stigmasterol are also likely to contribute to its chemopreventive action.
This study compares the diagnostic utility of complexed prostate-specific antigen (cPSA), total P... more This study compares the diagnostic utility of complexed prostate-specific antigen (cPSA), total PSA (tPSA) and their ratios with free PSA (fPSA) for benign prostatic hyperplasia (BPH) and prostate cancer. This is though to be the first study to evaluate cPSA in the ethnic population of Saudi Arabia. Serum samples were collected from 54 patients (aged over 50) and assayed for tPSA, cPSA and fPSA. Thirty-five patients were histologically and clinically proven to have BPH and 19 patients were proven to have cancer. Sensitivity, specificity and ROC curves were calculated. With a cPSA cut-off of 4 ng/mL the sensitivity was 79%, the specificity was 34%, and the positive and negative predictive values (PPV and NPV) were 39% and 75%, respectively. At the same cut-off for tPSA, the sensitivity was 84%, the specificity was 29%, and the PPV and NPV were 39 and 77%, respectively. The sensitivity for both tests was lower at a cut-off of 20 ng/mL but the specificity increased to 77% for cPSA and ...
This study aims to investigate whether the SNPs of CXCR1 gene, could predict the likelihood of vi... more This study aims to investigate whether the SNPs of CXCR1 gene, could predict the likelihood of viral persistence and/or disease progression. We investigated the association of two different SNPs (rs2234671, and rs142978743) in 598 normal healthy controls and 662 HBV patients from a Saudi ethnic population. The HBV patients were categorized into inactive carriers (n = 428), active carriers (n = 162), cirrhosis (n = 54) and Cirrhosis-HCC (n = 18) sub-groups. Genetic variants in CXCR1 were determined by polymerase chain reaction (PCR)-based DNA direct sequencing. The frequency of the risk allele 'C' for the SNP, rs2234671 was found to be insignificant when the patient group was compared to the uninfected control group, however, a significant distribution of the allele 'C' of rs2234671 was observed among active HBV carriers + cirrhosis + cirrhosis - HCC vs. inactive HBV carriers with an OR = 1.631 (95% C.I. 1.016-2.616) and p = 0.032. However, no significant association ...
Recently we have reported potent anti-cancer actions of various steroidal Na(+)/K(+) ATPase inhib... more Recently we have reported potent anti-cancer actions of various steroidal Na(+)/K(+) ATPase inhibitors in multiple cell lines. Furthermore, the most powerful compound identified in this study, the 3-[(R)-3-pyrrolidinyl]oxime derivative (3-R-POD), was highly effective in various tumor cell lines in vitro, and exhibited significant tumor growth inhibition in prostate and lung xenografts in vivo. In the present study we have addressed the molecular mechanisms implicated in the anti-cancer actions of 3-R-POD. We report here that 3-R-POD induces strong apoptotic responses in A549 lung- and in DU145 prostate- cancer cells. These effects are accompanied by significant upregulation of caspase-3 activity. Focussing on A549 cells, we further demonstrate late downregulation of BCL-2- and upregulation of c-Fos- gene transcription. In addition, the steroidal Na(+)/K(+) ATPase inhibitor induced late de-phosphorylation of Focal Adhesion Kinase (FAK) and activation of p38 MAPK. Our findings suggest...
Chorein, a protein supporting activation of phosphoinositide 3 kinase (PI3K), participates in the... more Chorein, a protein supporting activation of phosphoinositide 3 kinase (PI3K), participates in the regulation of actin polymerization and cell survival. A loss of function mutation of the chorein encoding gene VPS13A (vacuolar protein sorting-associated protein 13A) leads to chorea-acanthocytosis (ChAc), a neurodegenerative disorder with simultaneous erythrocyte akanthocytosis. In blood platelets chorein deficiency has been shown to compromise expression of vesicle-associated membrane protein 8 (VAMP8) and thus degranulation. The present study explored whether chorein is similarly involved in VAMP8 expression and dopamine release of pheochromocytoma (PC12) cells. Chorein was down-regulated by silencing in PC12 cells. Transmission electron microscopy was employed to quantify the number of vesicles, RT-PCR to determine transcript levels, Western blotting to quantify protein expression and ELISA to determine dopamine release. Chorein silencing significantly reduced the number of vesicles, VAMP8 transcript levels and VAMP8 protein abundance. Increase of extracellular K+ from 5 mM to 40 mM resulted in marked stimulation of dopamine release, an effect significantly blunted by chorein silencing. Chorein deficiency down-regulates VAMP8 expression, vesicle numbers and dopamine release in pheochromocytoma cells.
Mutation Research/Genetic Toxicology and Environmental Mutagenesis, Jun 15, 2012
The mutagenic potential of titanium dioxide nanoparticles (TiO2-NPs) of an average size 30.6 nm w... more The mutagenic potential of titanium dioxide nanoparticles (TiO2-NPs) of an average size 30.6 nm was investigated using β-galactosidase (lacZ) gene complementation in plasmid pUC19/lacZ− Escherichia coli DH5α system. Plasmid pUC19 was treated with varying concentrations of TiO2-NPs and allowed to transfect the CaCl2-induced competent DH5α cells. The data revealed loss in transformation efficiency of TiO2-NPs treated plasmids as compared to untreated plasmid DNA in DH5α host cells. Induction of multiple mutations in ...
Annals of Hepatology Official Journal of the Mexican Association of Hepatology, Mar 1, 2013
This study aims to investigate whether the SNPs of CXCR1 gene, could predict the likelihood of vi... more This study aims to investigate whether the SNPs of CXCR1 gene, could predict the likelihood of viral persistence and/or disease progression. We investigated the association of two different SNPs (rs2234671, and rs142978743) in 598 normal healthy controls and 662 HBV patients from a Saudi ethnic population. The HBV patients were categorized into inactive carriers (n = 428), active carriers (n = 162), cirrhosis (n = 54) and Cirrhosis-HCC (n = 18) sub-groups. Genetic variants in CXCR1 were determined by polymerase chain reaction (PCR)-based DNA direct sequencing. The frequency of the risk allele 'C' for the SNP, rs2234671 was found to be insignificant when the patient group was compared to the uninfected control group, however, a significant distribution of the allele 'C' of rs2234671 was observed among active HBV carriers + cirrhosis + cirrhosis - HCC vs. inactive HBV carriers with an OR = 1.631 (95% C.I. 1.016-2.616) and p = 0.032. However, no significant association was observed for rs142978743 when the distribution of risk allele was analyzed among the different patient groups (i.e. inactive carriers, active carriers, cirrhosis and HCC). Furthermore, the most common haplotype, Haplo-1 (AG), was found to have an insignificant frequency distribution between HBV cases and controls, while the same haplotype was found to be significantly distributed when active carriers + cirrhosis + cirrhosis - HCC patients were compared to inactive HBV carriers with a frequency of 0.938 and p = 0.0315. Haplo-2 (AC) was also found to be significantly associated with a frequency of 0.058 and p = 0.0163. The CXCR1 polymorphism, rs2234671 was found to be associated with chronic HBV infection and may play a role in disease activity.
Sodium potassium pump (Na+/K+ ATPase) is a validated pharmacological target for the treatment of ... more Sodium potassium pump (Na+/K+ ATPase) is a validated pharmacological target for the treatment of various cardiac conditions. Recent published data with Na+/K+ ATPase inhibitors suggest a potent anti-cancer action of these agents in multiple indications. In the present study, we focus on istaroxime, a Na+/K+ ATPase inhibitor that has shown favorable safety and efficacy properties in cardiac phase II clinical trials. Our experiments in 22 cancer cell lines and in prostate tumors in vivo proved the strong anti-cancer action of this compound. Istaroxime induced apoptosis, affected the key proliferative and apoptotic mediators c-Myc and caspase-3 and modified actin cystoskeleton dynamics and RhoA activity in prostate cancer cells. Interestingly, istaroxime was capable of binding to mAR, a membrane receptor mediating rapid, non-genomic actions of steroids in prostate and other cells. These results support a multi-level action of Na+/K+ ATPase inhibitors in cancer cells and collectively validate istaroxime as a strong re-purposing candidate for further cancer drug development.
Chromium oxide nanoparticles (Cr2O3NPs) are widely used in polymers and paints. In the present st... more Chromium oxide nanoparticles (Cr2O3NPs) are widely used in polymers and paints. In the present study, we aimed to determine the toxicity of Cr2O3NPs in murine fibrosarcoma (L929) cells. The cytotoxicity of Cr2O3NPs was measured by MTT and neutral red uptake assays; Cr2O3NPs had significant cytotoxic effects on L929 cells. Enhancement of intracellular reactive oxygen species was observed in L929 cells after exposure to Cr2O3NPs. Cr2O3NPs produced caspase-3, indicating that exposure to Cr2O3NPs induced apoptosis. After exposure to Cr2O3NPs, the cellular glutathione level decreased and lipid peroxidation, superoxide dismutase, and catalase increased in a dose- and time-dependent manner. By using single-cell gel tests, we also observed increased DNA damage in a Cr2O3NP exposure-duration- and dose-dependent fashion. Cell toxicity and DNA damage may be useful biomarkers for determining the safety of Cr2O3NPs in human and animal health.
FGF23 regulates renal phosphate and vitamin D metabolism. Loss of FGF23 results in massive calcif... more FGF23 regulates renal phosphate and vitamin D metabolism. Loss of FGF23 results in massive calcification and rapid aging. FGF23 production is stimulated by 1,25(OH)2 D3 and NFκB signaling. Here, we report that treatment of UMR106 osteoblast-like cells with 1,25(OH)2 D3 , inducing Fgf23 transcription, resulted in actin polymerization which was blocked by NFκB inhibitor wogonin. Interestingly, 1,25(OH)2 D3 -induced Fgf23 gene transcription was abolished by the actin microfilament-disrupting agent cytochalasin B, as well as by the inhibition of actin-regulating Rac1/PAK1 signaling. Our results provide strong evidence that actin redistribution regulated by the Rac1/PAK1 pathway participates in 1,25(OH)2 D3 -induced Fgf23 gene transcription. This article is protected by copyright. All rights reserved.
Barium oxide nanoparticles (BaONPs) are an important industrial compound and are widely used in p... more Barium oxide nanoparticles (BaONPs) are an important industrial compound and are widely used in polymers and paints. In this study, apoptotic and genotoxic effects of BaONPs in mouse embryonic fibroblast (L929) cells were determined by using single-cell gel test. In vitro cytotoxicity assays were performed to assess BaONPs' toxicity in L929 cells. Mild cytotoxicity was observed in L929 cells due to BaONPs. BaONPs increased lipid peroxidation, catalase, and superoxide dismutase levels and lowered glutathione levels in L929 cells. This was accompanied by concomitant generation of reactive oxygen species and activation of caspase-3 in BaONPs-treated L929 cells. On the other hand, when we exposed L929 cells to BaONPs for 24 and 48 hours (comet assay), there was a duration- and dose-dependent increase in DNA impairment detected in the single-cell gel test. Thus, BaONPs exhibit genotoxic and apoptotic effects in L929 cells, most likely due to initiation of oxidative damage.
Quercetol is a polyphenolic molecule present in vegetables and fruits, and is beneficial to human... more Quercetol is a polyphenolic molecule present in vegetables and fruits, and is beneficial to human and animal health. The current work aimed to test cytotoxic and apoptotic effects of quercetol on HepG2 cells. Quercetol was isolated from Ocimum sanctum and characterized by gas chromatography-tandom mass spectrometry (GC-MS/MS), nuclear magnetic resonance spectroscopy, and Fourier transform infrared spectroscopy. Quercetol (50-600 μg/mL) was examined for cytotoxic activity by tetrazolium salt and neutral red uptake tests and comet assay for genotoxicity, using HepG2 cells, over 24 hours. Data from 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide and neutral red uptake tests demonstrated quercetol-induced cytotoxicity in HepG2 cells in a concentration-dependent manner. With 4',6-diamidino-2-phenylindole staining, a significant induction of chromosomal condensation was observed at 300 μg/mL of quercetol. DNA fragmentation analysis showed that quercetol produced cell death in HepG2 cells in a concentration-dependent manner. Thus, our study suggests that an environmentally relevant concentration of quercetol, which was a chemically standardized extract from O. sanctum, induced cell death and DNA damage in HepG2 cells.
This study compares the diagnostic utility of complexed prostate-specific antigen (cPSA), total P... more This study compares the diagnostic utility of complexed prostate-specific antigen (cPSA), total PSA (tPSA) and their ratios with free PSA (fPSA) for benign prostatic hyperplasia (BPH) and prostate cancer. This is though to be the first study to evaluate cPSA in the ethnic population of Saudi Arabia. Serum samples were collected from 54 patients (aged over 50) and assayed for tPSA, cPSA and fPSA. Thirty-five patients were histologically and clinically proven to have BPH and 19 patients were proven to have cancer. Sensitivity, specificity and ROC curves were calculated. With a cPSA cut-off of 4 ng/mL the sensitivity was 79%, the specificity was 34%, and the positive and negative predictive values (PPV and NPV) were 39% and 75%, respectively. At the same cut-off for tPSA, the sensitivity was 84%, the specificity was 29%, and the PPV and NPV were 39 and 77%, respectively. The sensitivity for both tests was lower at a cut-off of 20 ng/mL but the specificity increased to 77% for cPSA and ...
This study was conducted to investigate the ultrastructural changes in the liver and kidney of mi... more This study was conducted to investigate the ultrastructural changes in the liver and kidney of mice induced by a single dose of Cadmium chloride. A group of mice (25) were injected with a single intraperitoneal dose of Cadmium chloride (2 mg/ kg) body weight. After 4 weeks, all treated and untreated animals were killed, the live and kidney were removed and processed for transmission electron microscopy. Ultrastructure of liver tissues showed increase in the number of lysosomal structure and oil droplets. Also changes in the mitochondrial structures and enlarge in Kupffer cells were observed. Ultrastructure of kidney tubule cells showed cellular necrosis, disappearing of microvilli and changes in the mitochondria and nucleus. Basing upon the results of electron microscopy investigation, it was possible to conclude that a single PI dose of cadmium chloride (2mg/kg) can induce ultrastructural changes in both liver and kidney tissues
Journal of Environmental Pathology, Toxicology and Oncology, 2015
This study aims to investigate whether asiaticoside, a triterpene glycoside, can afford protectio... more This study aims to investigate whether asiaticoside, a triterpene glycoside, can afford protection to DNA from alterations induced by gamma radiation under in vitro, ex vivo, and in vivo conditions. In vitro studies were done on plasmid pBR322 DNA, ex vivo studies were done on cellular DNA of human peripheral blood leukocytes, and in vivo investigations were conducted on cellular DNA of spleen and bone marrow cells of mice exposed to whole-body gamma radiation. The supercoiled form of the plasmid pBR322 DNA upon exposure to the radiation was converted into relaxed open circular form due to induction of strand breaks. Presence of asiaticoside along with the DNA during irradiation prevented the relaxation of the supercoiled form to the open circular form. When human peripheral blood leukocytes were exposed to gamma radiation, the cellular DNA suffered strand breaks as evidenced by the increased comet parameters in an alkaline comet assay. Asiaticoside, when present along with blood during irradiation ex vivo, prevented the strand breaks and the comet parameters were closer to that of the controls. Whole-body exposure of mice to gamma radiation resulted in a significant increase in comet parameters of DNA of bone marrow and spleen cells of mice as a result of radiation-induced strand breaks in DNA. Administration of asiaticoside prior to whole-body radiation exposure of the mice prevented this increase in radiation-induced increase in comet parameters, which could be the result of protection to DNA under in vivo conditions of radiation exposure. Thus, it can be concluded from the results that asiaticoside can offer protection to DNA from radiation-induced alterations under in vitro, ex vivo, and in vivo conditions.
Stigmasterol (99.9% pure) was isolated from Azadirachta indica and its chemopreventive effect on ... more Stigmasterol (99.9% pure) was isolated from Azadirachta indica and its chemopreventive effect on 7,12-dimethylbenz[a]anthracene (DMBA)-induced skin cancer was investigated in Swiss albino mice. Skin tumors were induced by topical application of DMBA and promoted by croton oil. To assess the chemopreventive potential of stigmasterol, it was orally administered at a concentration of 200 mg/kg and 400 mg/kg three times weekly for 16 weeks. Reduction in tumor size and cumulative number of papillomas were seen as a result of treatment with stigmasterol. The average latency period was significantly increased as compared with the carcinogen-treated control. Stigmasterol induced a significant decrease in the activity of serum enzymes, such as aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, and bilirubin as compared with the control. Stigmasterol significantly increased glutathione, superoxide dismutase, and catalase as compared with the control. Elevated levels of lipid peroxide and DNA damage in the control group were significantly inhibited by administration of stigmasterol. From the present study, it can be inferred that stigmasterol has chemopreventive activity in an experimental model of cancer. This chemopreventive activity may be linked to the oxidative stress of stigmasterol. The antigenotoxic properties of stigmasterol are also likely to contribute to its chemopreventive action.
This study compares the diagnostic utility of complexed prostate-specific antigen (cPSA), total P... more This study compares the diagnostic utility of complexed prostate-specific antigen (cPSA), total PSA (tPSA) and their ratios with free PSA (fPSA) for benign prostatic hyperplasia (BPH) and prostate cancer. This is though to be the first study to evaluate cPSA in the ethnic population of Saudi Arabia. Serum samples were collected from 54 patients (aged over 50) and assayed for tPSA, cPSA and fPSA. Thirty-five patients were histologically and clinically proven to have BPH and 19 patients were proven to have cancer. Sensitivity, specificity and ROC curves were calculated. With a cPSA cut-off of 4 ng/mL the sensitivity was 79%, the specificity was 34%, and the positive and negative predictive values (PPV and NPV) were 39% and 75%, respectively. At the same cut-off for tPSA, the sensitivity was 84%, the specificity was 29%, and the PPV and NPV were 39 and 77%, respectively. The sensitivity for both tests was lower at a cut-off of 20 ng/mL but the specificity increased to 77% for cPSA and ...
This study aims to investigate whether the SNPs of CXCR1 gene, could predict the likelihood of vi... more This study aims to investigate whether the SNPs of CXCR1 gene, could predict the likelihood of viral persistence and/or disease progression. We investigated the association of two different SNPs (rs2234671, and rs142978743) in 598 normal healthy controls and 662 HBV patients from a Saudi ethnic population. The HBV patients were categorized into inactive carriers (n = 428), active carriers (n = 162), cirrhosis (n = 54) and Cirrhosis-HCC (n = 18) sub-groups. Genetic variants in CXCR1 were determined by polymerase chain reaction (PCR)-based DNA direct sequencing. The frequency of the risk allele 'C' for the SNP, rs2234671 was found to be insignificant when the patient group was compared to the uninfected control group, however, a significant distribution of the allele 'C' of rs2234671 was observed among active HBV carriers + cirrhosis + cirrhosis - HCC vs. inactive HBV carriers with an OR = 1.631 (95% C.I. 1.016-2.616) and p = 0.032. However, no significant association ...
Recently we have reported potent anti-cancer actions of various steroidal Na(+)/K(+) ATPase inhib... more Recently we have reported potent anti-cancer actions of various steroidal Na(+)/K(+) ATPase inhibitors in multiple cell lines. Furthermore, the most powerful compound identified in this study, the 3-[(R)-3-pyrrolidinyl]oxime derivative (3-R-POD), was highly effective in various tumor cell lines in vitro, and exhibited significant tumor growth inhibition in prostate and lung xenografts in vivo. In the present study we have addressed the molecular mechanisms implicated in the anti-cancer actions of 3-R-POD. We report here that 3-R-POD induces strong apoptotic responses in A549 lung- and in DU145 prostate- cancer cells. These effects are accompanied by significant upregulation of caspase-3 activity. Focussing on A549 cells, we further demonstrate late downregulation of BCL-2- and upregulation of c-Fos- gene transcription. In addition, the steroidal Na(+)/K(+) ATPase inhibitor induced late de-phosphorylation of Focal Adhesion Kinase (FAK) and activation of p38 MAPK. Our findings suggest...
Iron oxide (Fe3O4) nanoparticles (IONPs) have received much attention for their utility in biomed... more Iron oxide (Fe3O4) nanoparticles (IONPs) have received much attention for their utility in biomedical applications such as magnetic resonance imaging, drug delivery and hyperthermia. Recent studies reported that IONPs induced cytotoxicity in mammalian cells. However, little is known about the genotoxicity of IONPs following exposure to human cells. In this study, we investigated the cytotoxicity, oxidative stress and genotoxicity of IONPs in two human cell lines; skin epithelial A431 and lung epithelial A549. Prepared IONPs were polygonal in shape with a smooth surface and had an average diameter of 25 nm. IONPs (25-100 μg/ml) induced dose-dependent cytotoxicity in both types of cells, which was demonstrated by cell viability (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide) and lactate dehydrogenase leakage assays. IONPs were also found to induce oxidative stress in a dose-dependent manner, evident by depletion of glutathione and induction of reactive oxygen species (ROS) and lipid peroxidation. Comet assay revealed that level of DNA damage was higher with concentration of IONPs in both types of cells. Quantitative real-time PCR analysis showed that following the exposure of cells to IONPs, the expression levels of mRNA of caspase-3 and caspase-9 genes were higher. We also observed the higher activity of caspase-3 and caspase-9 enzymes in IONPs treated cells. Moreover, western blot analysis showed that protein expression level of cleaved caspase-3 was up-regulated by IONPs in both types of cells. Taken together, our data demonstrates that IONPs have potential to induce genotoxicity in A431 and A549 cells, which is likely to be mediated through ROS generation and oxidative stress. This study suggests that genotoxic effects of IONPs should be further investigated at in vivo level.
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