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MRN-100, An Iron-Based Compound, Possesses Anti-HIV Activity In Vitro
Mamdooh Ghoneum,Magda Shaheen
Evidence-Based Complementary and Alternative Medicine , 2010, DOI: 10.1093/ecam/nen019
Abstract: We examined the in vitro anti-human immunodeficiency virus (HIV) activity of MRN-100, an iron-based compound derived from bivalent and tervalent ferrates. MRN-100 action against HIV-1 (SF strain) was tested in primary cultures of peripheral blood mononuclear cells (MNC) by analyzing p24 antigen production and percent survival of MNC infected with HIV. MRN-100 at a concentration of 10% (v/v) inhibited HIV-1 replication in 11 out of 14 samples (79%). The percentage of suppression of p24 antigen was ?12.3 to 100% at 10 days post-treatment. MRN-100 also exhibited a significant protective effect in the survival of HIV-1-infected MNC. MNC survival post-treatment was dose dependent, 70.4% ± 8.4, 83.6% ± 10.7 and 90% ± 11.4, at concentrations 2.5, 5 and 10% (v/v), respectively, as compared with 53% ± 4 for HIV-1-infected MNC without treatment. The effect was detected as early as 4 days and continued up to 11 days. Treatment with MRN-100 caused no significant change in proliferative response of MNC alone or cocultured with different mitogens: PHA and Con-A (activators of T cell function) and PWM (activator of CD4+ T cell-dependent B cells). We concluded that MRN-100 possesses anti-HIV activity in vitro and without an increase in lymphocyte proliferation, MRN-100 may be a useful agent for treating patients with acquired immunodeficiency syndrome.
An iron-based beverage, HydroFerrate fluid (MRN-100), alleviates oxidative stress in murine lymphocytes in vitro
Mamdooh Ghoneum, Motohiro Matsuura, Sastry Gollapudi
Nutrition Journal , 2009, DOI: 10.1186/1475-2891-8-18
Abstract: Splenic lymphocytes from mice were cultured in the presence or absence of MRN-100 for 2 hrs and were subsequently exposed to hydrogen peroxide (H2O2) at a concentration of 25 μM for 14 hrs. Percent cell death was examined by flow cytometry and trypan blue exclusion. The effect of MRN-100 on Bcl-2 and Bax protein levels was determined by Western blot.Results show, as expected, that culture of splenic cells with H2O2 alone results in a significant increase in cell death (apoptosis) as compared to control (CM) cells. In contrast, pre-treatment of cells with MRN-100 followed by H2O2 treatment results in significantly reduced levels of apoptosis.In addition, MRN-100 partially prevents H2O2-induced down-regulation of the anti-apoptotic molecule Bcl-2 and upregulation of the pro-apoptotic molecule Bax.Our findings suggest that MRN-100 may offer a protective effect against oxidative stress-induced apoptosis in lymphocytes.Oxidative stress represents the imbalance between the cellular production of oxidants and the capacity of cellular antioxidant defenses to scavenge these oxidants. It is produced in cells by oxygen-derived species which include free radicals and peroxides; it is also produced at a low level by normal aerobic metabolism and nutritional deficiency in trace metal [1,2]. Increasing evidence indicates that oxidative stress is a major inducer of cell death [3]. In this process some of the reactive oxygen species (such as superoxide) are converted into hydrogen peroxide which can cause controlled apoptotic cell death [4,5].Oxidative stress is associated with many diseases, including chronic inflammation, arteriosclerosis, diabetes, stroke, Alzheimer's and Parkinson's diseases, and aging [6-8]. In addition, nutritional deficiencies like lack of iron have been shown to induce oxidative stress [9,10] and currently affect over 2 billion people worldwide. Thus far, the ability of iron to protect against oxidative stress has only been studied to a limited extent [11].I
Identification of Circulating Endothelial Cells in Peripheral Blood
Mamdooh Gari
Research Journal of Medical Sciences , 2012, DOI: 10.3923/rjmsci.2011.262.268
Abstract: The Circulating Endothelial Cells (CECs) and their activated and resting subsets (aCECs and rCECs) represent extremely rare cell population that play important roles in vascular pathophysiology. Their number and function are modulated in several diseases involving vascular injury such as human tumors. Although, consensuses on the phenotypic definition of endothelial cells and on the optimal enumeration technique is lacking, the number of clinical studies that are based on the assessment of endothelial cells the types of analytical methods that are employed are rapidly expanding. The goal of the current study was to develop a rapid and sensitive flow cytometric method to quantify and characterize CECs (their subsets and the apoptotic fraction of cells. In total, 75 peripheral blood samples were collected from normal donors and were analyzed with a six-color flow cytometric technique for the simultaneous analysis of the cell phenotypes of CECs and circulating progenitors cells using the following monoclonal antibodies: CD146, CD34, CD45, CD106 and CD133. In addition, the samples were analyzed with using the gating strategy. Apoptotic CECs and dead cells were detected using Annexin V and 7-amino-actinomycin D staining, respectively. The results show that the described technique is a reliable tool to increase the knowledge of endothelial cell biology and can be easily applied to the study of many pathological conditions.
JAK2 Mutations in Chronic Myeloproliferative Neoplasm; Towards the Application of Personalized Treatments for Saudi Patients  [PDF]
Mamdooh Gari, Fatin Al-Sayes, Farid Ahmed, Abdul Ali Peerzada, Adel Abuzenadah, Lubna S. Mira, Mohammed Al-Qahtani, Ashraf Dallol, Adeel Chaudhary, Mohammed Mahnashi, Ghazi Damanhouri
Open Journal of Blood Diseases (OJBD) , 2012, DOI: 10.4236/ojbd.2012.22004
Abstract: The chronic myeloproliferative neoplasms (CMPN) are a group of clonal hematopoietic stem cell disorders in which large numbers of red blood cells, white blood cells, or platelets grow and spread excess in the bone marrow and the pe- ripheral blood. Cytogenetic analysis of the t (9:22) and molecular detection of BCR/ABL is the main diagnostic criteria in Philadelphia positive CMPN (CML). The identification of non-receptor tyrosine kinase JAK2 mutations (exon 14 JAK2 V617F and exon 12) have significantly contributed to our understanding of the molecular mechanisms in the pathogenesis of Philadelphia negative CMPN such as polycythemia vera (PV), essential thrombocythemia (ET) and primary myelofibrosis (PMF) patients. According to the revised WHO classification, JAK2 mutation is considered as a major diagnostic and clonal marker in Philadelphia negative CMPN which will play a major role in designing personal- ized treatments for the disease. JAK2 V617F mutation frequency is unknown in Saudi Arabia. Therefore, investigation of the JAK2 V617F mutation was carried out in DNA samples from 78 peripheral blood specimens corresponding to patients with polycythemia vera (PV) (n = 11), Chronic myeloid leukemia (CML) (n = 45), essential thrombocythemia (ET) (n = 10), idiopathic myelofibrosis (MF) (n = 12). We used polymerase chain reaction and direct DNA sequencing to detect the JAK2 mutation. Overall, the incidence of the JAK2 V617F mutation was 91% in PV, 40% in ET, and 25% in MF. This approach proved to be reliable and more sensitive in detecting the mutation. Two essential findings arose from our study. First, this technique could be carried out with DNA samples, even partially degraded, from routinely processed BM or peripheral blood specimens. Second, after correlation with morphological features, it turned out that the characteristics of the megakaryocytes were more specific than the mutational status of JAK2 in characterizing ET and PMF. Concerning PV, as expected, the incidence of the JAK2 mutation was higher, but the morphological criteria were misleading in some cases, strongly suggesting that the combination of both morphology and molecular data would enable the characterization of virtually all cases. JAK2 V617F mutation frequency along with accurate morphological characterization is very reliable tool in diagnosing and classifying CMPN in Saudi patients.
Use of black pepper (Piper nigrum) as feed additive in broilers diet
Galib A. M. Al-Kassie,,Mamdooh A. M. Al-Nasrawi,,Saba J. Ajeena
Research Opinions in Animal & Veterinary Sciences , 2011,
Abstract: This study was conducted to determine the performance of broilers fed diets with black pepper (Piper nigrum). A total of 250 (Rose 308) day old chicks were used in this study. Five levels of black pepper at the rate of 0.00%, 0.25%, 0.50%, 0.75% and 1% were incorporated into the basal diet of broilers for six weeks. The Results revealed that the inclusion of black pepper at the levels of 0.50%, 0.75% and 1% in the diets improved body weight gain, feed intake and conversion ratio. At the same time the black pepper of 0.50 %, 0.75% and 1% depressed the cholesterol, Hb, RBC and H/L ratio concentration. It was concluded that the use of black pepper as feed additive at 0.50%, 0.75% and 1% enhanced the overall performance of broiler chicks.
Wagieh k.Baiomy; Abdel-mawgood Anas; Mamdooh Ghaly; Ashraf M. Moustafa
Egyptian Journal of Hospital Medicine , 2009,
Abstract: Background: The present work was based on the evaluation of histological, histochemical, and quantitative study on the adrenal medulla of the white albino rat in the different post natal age period. Material and methods:Sixty male albino rats were used in this study. The rats were classified to 4 main groups as follows: - Group one : One week old albino rats. - Group two: One month old albino rats. - Group three: Three months old albino rats. - Group four: Senile rats. Three main parameters were performed in this study, the first was the study of the morphological changes in the adrenal medulla in the different postnatal age groups. The second was concerned with the histochemical studies while the last parameter was the quantitative studies on the gland volume as well as its cellular count. These three parameters were performed by using different staining techniques. Results: The results showed that medullary cells in the early age groups were arranged in non-differentiated groups and become more differentiated in the older age groups. Both reticular and elastic fibers in the older age groups showed a definite increase especially at the region of corticomedullary zone. The different types of chromaffin cells were more observed at the old age groups. The concentration of ascorbic acid granules was more marked in the senile group. The quantitative changes were in the form of increased medullary volume especially in the old age. The number of chromaffin cells as well as the concentration of ascorbic acid contents was more noticed in the old age group. Conclusions: The differentiation of both divisions of the adrenal gland was not noticed in the early age groups. Cellular and fibrous differentiations were more seen in older age groups which may reflects an idea about the degree of gland maturation
Five novel glucose-6-phosphate dehydrogenase deficiency haplotypes correlating with disease severity
Dallol Ashraf,Banni Huda,Gari Mamdooh A,Al-Qahtani Mohammed H
Journal of Translational Medicine , 2012, DOI: 10.1186/1479-5876-10-199
Abstract: Background Glucose-6-phosphate dehydrogenase (G6PD, EC deficiency is caused by one or more mutations in the G6PD gene on chromosome X. An association between enzyme levels and gene haplotypes remains to be established. Methods In this study, we determined G6PD enzyme levels and sequenced the coding region, including the intron-exon boundaries, in a group of individuals (163 males and 86 females) who were referred to the clinic with suspected G6PD deficiency. The sequence data were analysed by physical linkage analysis and PHASE haplotype reconstruction. Results All previously reported G6PD missense changes, including the AURES, MEDITERRANEAN, A-, SIBARI, VIANGCHAN and ANANT, were identified in our cohort. The AURES mutation (p.Ile48Thr) was the most common variant in the cohort (30% in males patients) followed by the Mediterranean variant (p.Ser188Phe) detectable in 17.79% in male patients. Variant forms of the A- mutation (p.Val68Met, p.Asn126Asp or a combination of both) were detectable in 15.33% of the male patients. However, unique to this study, several of such mutations co-existed in the same patient as shown by physical linkage in males or PHASE haplotype reconstruction in females. Based on 6 non-synonymous variants of G6PD, 13 different haplotypes (13 in males, 8 in females) were identified. Five of these were previously unreported (Jeddah A, B, C, D and E) and were defined by previously unreported combinations of extant mutations where patients harbouring these haplotypes exhibited severe G6PD deficiency. Conclusions Our findings will help design a focused population screening approach and provide better management for G6PD deficiency patients.
Sunitinib Reduces Acute Myeloid Leukemia Clonogenic Cells in Vitro and Has Potent Inhibitory Effect on Sorted AML ALDH+ Cells  [PDF]
Asad M. Ilyas, Youssri Ahmed, Mamdooh Gari, Mohammed H. Alqahtani, Taha A. Kumosani, Abdulrahman L. Al-Malki, Khalid O. Abualnaja, Saad H. S. Albohairi, Adeel G. A. Chaudhary, Farid Ahmed
Open Journal of Blood Diseases (OJBD) , 2016, DOI: 10.4236/ojbd.2016.61003
Abstract: Sunitinib is an orally administered, multi-target tyrosine kinase inhibitor that has been approved by the FDA for the treatment of renal cell carcinoma and imatinib resistant gastro-intestinal tumors. Anti-leukemic activity of sunitinib has been examined in early clinical trials with limited success. However, recent trials on acute myeloid leukemia (AML) patients carrying FLT3 mutations have shown promising results. Effects of sunitinib on leukemic clonogenic cells and potential leukemic stem cells have not been examined so far. We analyzed the anti-proliferative and apoptotic properties of sunitinib on AML-derived cell lines. We also tested the effect of sunitinib on AML patient derived clonogenic cells (AML-CFC), as well as flow-sorted potential leukemic progenitors. Peripheral blood or bone marrow samples were obtained from newly diagnosed AML patients and flow sorted for CD34+ CD133+ or ALDH+ cells. Umbilical cord blood derived CD34+ cells were used as normal controls. Sunitinib induced growth arrest and apoptosis in AML derived cell lines. In addition, 7 μM sunitinib induced 75% reduction of AML-CFC as compared to DMSO treated control (±6.79%; n = 4). In contrast, 7 μM sunitinib treatment of umbilical cord blood derived normal CD34+ cells showed 29% reduction in AML-CFC (±6.77%; n = 5). Treatment of ALDH+ cells sorted from 2 AML cases and CD34+ CD133+ cells from one patient showed reduction of AML-CFC on treatment with sunitinib. Our study highlighted a potent anti-proliferative and proapoptotic effect of sunitinib on AML cell lines, AML patient derived clonogenic cells and potential leukemic stem cells.
Detection of FLT3 Oncogene Mutations in Acute Myeloid Leukemia Using Conformation Sensitive Gel Electrophoresis
Mamdooh Gari,Adel Abuzenadah,Adeel Chaudhary,Mohammed Al-Qahtani,Huda Banni,Waseem Ahmad,Fatin Al-Sayes,Sahira Lary,Ghazi Damanhouri
International Journal of Molecular Sciences , 2008, DOI: 10.3390/ijms9112194
Abstract: FLT3 (fms-related tyrosine kinase 3) is a receptor tyrosine kinase class III that is expressed on by early hematopoietic progenitor cells and plays an important role in hematopoietic stem cell proliferation, differentiation and survival. FLT3 is also expressed on leukemia blasts in most cases of acute myeloid leukemia (AML). In order to determine the frequency of FLT3 oncogene mutations, we analyzed genomic DNA of adult de novo acute myeloid leukemia (AML). Polymerase chain reaction (PCR) and conformation-sensitive gel electrophoresis (CSGE) were used for FLT3 exons 11, 14, and 15, followed by direct DNA sequencing. Two different types of functionally important FLT 3 mutations have been identified. Those mutations were unique to patients with inv(16), t(15:17) or t(8;21) and comprised fifteen cases with internal tandem duplication (ITD) mutation in the juxtamembrane domain and eleven cases with point mutation (exon 20, Asp835Tyr). The high frequency of the flt3 proto-oncogene mutations in acute myeloid leukemia AML suggests a key role for the receptor function. The association of FLT3 mutations with chromosomal abnormalities invites speculation as to the link between these two changes in the pathogenesis of acute myeloid leukemiaAML. Furthermore, CSGE method has shown to be a rapid and sensitive screening method for detection of nucleotide alteration in FLT3 gene. Finally, this study reports, for the first time in Saudi Arabia, mutations in the human FLT3 gene in acute myeloid leukemia AML patients.
Identification of Unique miRNA Biomarkers in Colorectal Adenoma and Carcinoma Using Microarray: Evaluation of Their Putative Role in Disease Progression
Kothandaraman Narasimhan,Kalamegam Gauthaman,Peter Natesan Pushparaj,Govindasamy Meenakumari,Adeel Gulzar Ahmed Chaudhary,Adel Abuzenadah,Mamdooh Abdullah Gari,Mohammed Al Qahtani,Jayapal Manikandan
ISRN Cell Biology , 2014, DOI: 10.1155/2014/526075
Abstract: MicroRNAs (miRNAs) are known to be dysregulated and play a key role in cancer progression. The present study aims to identify the miRNAs associated with colorectal adenoma and carcinoma to evaluate their role in tumor progression and metastasis using microarray. In silico analysis of miRNAs was performed on five different microarray data sets that represented the genes and miRNAs expressed in colorectal adenoma and carcinoma. We identified 10 different miRNAs that were common to both colorectal adenoma and carcinoma, namely, miR9, miR96, miR135b, miR137, miR147, miR182, miR183, miR196b, miR224, and miR503. Of these, miR135b and miR147 were significantly downregulated in colorectal adenoma but upregulated in carcinoma. In addition, we studied the gene expression profile associated with colorectal adenocarcinoma and identified three genes, namely, ZBED3, SLC10A3, and FOXQ1, that were significantly downregulated in colorectal adenoma compared to carcinoma. Interestingly, of all the miRNAs and genes associated with colorectal adenocarcinoma, the myoglobin (MB) gene was identified to be under the direct influence of miR135b, showing an inverse relationship between them in adenoma and carcinoma. Most of the identified miRNAs and associated genes are involved in signaling pathways of cell proliferation, angiogenesis, and metastasis. The present study has identified putative miRNA targets and their associated gene networks which could be used as potential biomarkers of colon adenocarcinoma. Moreover, the association of miR135b and MB gene is very unique and can be considered as a lead candidate for novel cancer therapeutics. 1. Introduction Colorectal cancer is one of the most common gastrointestinal cancers that shows a rising trend due to the dietary habits and lifestyle modifications. Accumulation of genetic and epigenetic aberrations predisposes the colonic epithelium to undergo gradual transformation with loss of cellular architecture and initiation of a benign adenoma, which subsequently develops into a malignant adenocarcinoma [1]. Successful screening methods have enabled early detection and hence reduction in the mortality rate associated with colorectal adenocarcinoma [2]. In addition to the oncogenes and the tumor suppressor genes involved in cancer initiation and progression, many other small molecules such as the oncoproteins, antisense RNAs, and microRNAs (miRNAs) are also implicated in cancer and its signaling mechanisms [3]. Among the small molecules involved in cancer progression and metastasis, the microRNAs (miRNAs) which are 21–25
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