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Article
Novel methods to design wild bacteriophages into highly lytic and therapeutic bacteriophages to extensively drug-resistant Mycobacterium tuberculosis
طرق حديثة وجديدة لتصميم بكتريوفاج ذو فعالية علاجية ومدمرة للبكتريا Mycobacterium tuberculosis المقاومة

Author: Ahmed S. Abdulamir احمد صاحب عبد الامير
Journal: Journal of the Faculty of Medicine مجلة كلية الطب ISSN: 00419419 Year: 2016 Volume: 58 Issue: 3 Pages: 276-282
Publisher: Baghdad University جامعة بغداد

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Abstract

Background: The emergence of multiple drug resistant (MDR) Mycobacterium tuberculosis (M.TB) and extensive drug resistant (XDR) M.TB lay huge burden on TB endemic countries such as Iraq.Objectives: Bacteriophage (phage) therapy can be used as alternative approach to tackle this problem. Patients and methods: Forty isolates of M.TB were cultured from TB-positive sputum specimens with three ATCC strains. Phage passaging and biokinetic based techniques were used to optimize wild anti-M.TB phages. Three chemical, non genetic- designing techniques, tween-80, mycobacterial lysis buffer, and xyelen, were used to change the specificity of wild anti-TB phages towards phage-resistant target M.TB.Results: Five wild anti-M.TB phages were isolated and optimized. The optimization techniques were successful in enhancing plaques size, clarity, burst size, and infective ratio. Chemical designing techniques succeeded to reorient specificity of 6 phages to new host bacteria.Conclusions: phage designing opens door wide for endless future applications of phage-based therapy, biocontrol and diagnosis.

خلفية البحث: ان ظهور العديد من البكتريا المتفطرة السلية (M.TB) المقاومة (MDR)للعقاقير وظهور سلالات مقاومة لعقاقير واسعة النطاق (XDR) M.TB وضع عبئا كبيرا على الدول والموؤسسات الصحية مثل حالةالسل المتوطنة في العراق. الأهداف: علاجات العاثيات البكتيرية (فج) يمكن أن تستخدم كنهج بديل لمعالجة هذه المشكلة. المرضى وطرائق العمل: أربعون عزلة من M.TB زرعت من عينات البلغم السل الإيجابي مع ثلاث سلالات مقايسة من آي تي سي سي. تم استخعدام طريقة التهجين التسلسلي للعاثيات وتقنيات الأنشطة الحيوية لتحسين العاثيات البرية لمكافحة M.TB. ثلاثة مواد وتقنيات كيميائية وتقنيات تصميم غير ,وراثية استخدمت على العاثيات البكتيرية منها توين-80، محلول تحلل العزلة الفطرية، وxyelen، كانت تستخدم لتغيير خصوصية العاثيات البكتيرية المضادة لبكتيريا السل.. النتائج: تم عزل خمسة فاجات (عاثيات بكتيرية) لمكافحة M.TB البرية. ثلاث من تقنيات التصميم و التحسين كانت ناجحة حيث نجحت تقنيات التصميم الكيميائية لإعادة توجيه خصوصية 6 عاثيات بكتيرية الى بكتيريا مضيفة جديدةالاستنتاجات: ان تقنيات تصميم العاثيات البكتيرية يفتح الباب على مصراعيه للتطبيقات المستقبلية التي لا نهاية لها من العلاج والتشخيص والسيطرة البيئية المعتمدة على العاثيات البكتيرية.


Article
MOLECULAR CHARACTERIZATION OF THE ONCOGENIC POTENTIAL AND MECHANISMS OF CYTOMEGALOVIRUS INFECTING MRC-5 CELLS

Author: Ahmed S. Abdulamir احمد صاحب عبد الامير
Journal: IRAQI JOURNAL OF MEDICAL SCIENCES المجلة العراقية للعلوم الطبية ISSN: P16816579,E22244719 Year: 2016 Volume: 14 Issue: 4 Pages: 336-350
Publisher: Al-Nahrain University جامعة النهرين

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Abstract

Background:Human cytomegalovirus (HCMV), a member of herpes virus family, is associated with different cancers in humans, including breast and colorectal cancers.Objectives:To shed light on the possible oncogenic mechanisms exerted by HCMV on tissues at the genetic level.Methods: HCMV was propagated in medical research council-5 (MRC-5) cell line and cytopathic effects of HCMV were checked for confirming infection. Oncogenic mechanism that exerted by HCMV was screened using high-throughput real-time qPCR microarray to investigate the mRNA expression of 983 genes in 16 gene families associated with oncogenesis in human cells.Results :Twenty differentially expressed genes. Upregulated genes (SCGB1A1, MAP2K3, ITGB3, TGFB1, PTN, MMP2, AKT1, AKT2, BCL2L10, CKS1B, SERPINB4, CASP4, TOLLIP, BCL-2, SERPINB5) were mostly of oncogenic or oncopromoting activity, involved mainly in MAPK, PI3K/Akt, inflammation, and angiogenesis pathways while downregulated genes (STAT3, BAK1, BLM, RB1, and IGF2R) were mostly tumor suppressor, proapoptotic, and DNA damage repair genes.Conclusion:The current study revealed that HCMV might have an oncogenic or oncomodulating activity on normal infected cells by different intracellular pathways.Keywords: Cytomegalovirus, oncogenic viruses, mrc-5, Waffergen, SmartChip Real-Time PCR System.


Article
IN VITRO STUDY ON USING BACTERIOPHAGES IN THE TREATMENT OF PATHOGENIC ESCHERICHIA COLI IN IRAQ

Authors: Marwa B. Sabri مروه باسم صبري --- Ahmed S. Abdulamir احمد صاحب عبد الامير
Journal: IRAQI JOURNAL OF MEDICAL SCIENCES المجلة العراقية للعلوم الطبية ISSN: P16816579,E22244719 Year: 2014 Volume: 12 Issue: 2 Pages: 131-138
Publisher: Al-Nahrain University جامعة النهرين

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Abstract

Background:Bacteriophages, or phages, are virus-like agents that infect bacteria. Lytic phages can be used as biological antimicrobials and can kill bacteria.Objective:To formulate therapeutic phage cocktails able to overcome multiple-drug resistant Escherichia coli in Iraq.Methods:Escherichia coli were isolated from Iraqi hospitals and were characterized in terms of site of isolation, patient's age, sex, and disease. Antibiotics sensitivity test was used to evaluate antibiotics effectiveness. Accordingly, isolated bacteria were grouped in terms of resistance to antibiotics, infection type, and infection site. Wild phages specific to Escherichia coli were isolated from different areas. The isolated phages were optimized and their biokinetics were measured.Results:A total of 10 samples of Escherichia coli, 7 samples of them revealed specific phages. The formulated phage cocktail to Escherichia coli was shown to remarkably minimize the bacterial resistance to individual phages.Conclusion:Bacteriophage cocktails are useful to tackle the problem of MDR bacteria.Keywords:Multi-drug resistance, bacteriophage, phage therapy, Escherichia coli.


Article
STUDYING THE FREQUENCY OF METHICILLIN-RESISTANT STAPHYLOCOCCUS AUREUS THROUGH THE MOLECULAR DETECTION OF MECA

Authors: Rafeef Y. Rasheed رفيف يوسف رشيد --- Ahmed S. Abdulamir احمد صاحب عبد الامير --- Amir H. Raziq عامر هاني رزاق
Journal: IRAQI JOURNAL OF MEDICAL SCIENCES المجلة العراقية للعلوم الطبية ISSN: P16816579,E22244719 Year: 2014 Volume: 12 Issue: 3 Pages: 289-294
Publisher: Al-Nahrain University جامعة النهرين

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Abstract

Background:Methicillin-resistant Staphylococcus aureus is a major cause of serious nosocomial infections and it is very important to have a reliable test to detect these bacteria. mecA encodes the penicillin binding protein 2a, which is associated with staphylococcal methicillin resistance.Objective:The study was to determine the frequency of methicillin-resistant Staphylococcus aureus in different specimens from Iraqi patients and to genetically characterize and type the samples of methicillin- resistant Staphylococcus aureus through the detection of mec A gene.Methods:Sixty clinical isolates of Staphylococcus aureus were submitted to DNA extraction. Genomic DNA was submitted to conventional polymerase chain reaction assays, employing MR1-MR2 primers (primer set). The results were compared to the cefoxitin disks agar diffusion method.Results :Fifty seven of the sixty isolates showed positive results for mecA amplification while three isolates (5%) showed negative results for mecA gene.Conclusion:Good correlation between the mecA gene detection by PCR and the cefoxitin disk diffusion methods was obtained.Key words:mecA gene, Staphylococcus aureus, methicillin resistance Staphylococcus aureus


Article
8.EXTRACTION, PURIFICATION AND THERAPEUTIC USE OF BACTERIOPHAGE ENDOLYSIN AGAINST MULTI-DRUG RESISTANT PSEUDOMONAS AERUGINOSA: AN IN-VITRO AND IN-VIVO STUDY

Authors: Mohammed R. Ali محمد رزاق علي --- Shurooq R. Kadhim شروق ريس كاظم --- Ahmed S. Abdulamir احمد صاحب عبد الامير
Journal: IRAQI JOURNAL OF MEDICAL SCIENCES المجلة العراقية للعلوم الطبية ISSN: P16816579,E22244719 Year: 2018 Volume: 16 Issue: 4 Pages: 405-412
Publisher: Al-Nahrain University جامعة النهرين

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Abstract

Background: Persistent infection with multidrug resistant Pseudomonas aeruginosa (P. aeruginosa) represents a real problem for health care providers. Bacteriophage lytic enzymes or lysins are highly evolved molecules that have been specifically developed by phages to quickly and efficiently allow their progeny to be released from the host bacterium while destructing that bacterium.Objective: Isolation of endolysin from P. aeruginosa bacteriophages, and administering them systematically in vivo lab animal and measuring their therapeutic efficacy as well as evaluation of their biosafety.Methods: This study was performed from March 2015 – August 2017, during which 50 bacteriological samples of P. aeruginosa were collected, and examined for their antibiogram, then bacteriophage cocktails were done for 5 resistant strains of them. Endolysins were extracted from their corresponding bacteriophages and characterized. The enzymatic and antibacterial activities as well in vivo therapeutic efficiency of these enzymes were investigated.Results: This study showed that the extracted endolysin from these bacteriophages was effective in treating laboratory mice from bacteremia with P. aeruginosa and saving their lives when injected intraperitoneal.Conclusion: Endolysin can be extracted directly from their bacteriophages and used effectively in proper doses to treat bacteremia in mice.Keywords: Pseudomonas aeruginosa, bacteriophage, endolysin, MDRCitation: Ali MR, Kadhim SR, Abdulamir AS. Extraction, purification and therapeutic use of bacteriophage endolysin against multi-drug resistant Pseudomonas aeruginosa: An in-vitro and in-vivo study. Iraqi JMS. 2018; 16(4): 405-412. doi: 10.22578/IJMS.16.4.8


Article
THE SEROPOSITIVITY OF PARVOVIRUS B19 AMONG KIDNEY TRANSPLANT RECIPIENTS

Authors: Zainab A. Hlail زينب علي هليل --- Ahmed S. Abdulamir احمد صاحب عبد الامير --- Ali J.H. Al-Saedi علي جاسم الساعدي
Journal: IRAQI JOURNAL OF MEDICAL SCIENCES المجلة العراقية للعلوم الطبية ISSN: P16816579,E22244719 Year: 2017 Volume: 15 Issue: 1 Pages: 88-93
Publisher: Al-Nahrain University جامعة النهرين

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Abstract

Background: Parvovirus B19 (PV-B19) is a single strand DNA virus that is responsible for causing several diseases in humans. Parvovirus B19 induced persistent anemia or/and pancytopenia or/and reticulocyteopenia in renal transplant recipients (RTRs).Objective:To find out any association between PV-B19 reactivation / primary infections with abnormal renal function tests, and post-transplantation period in renal transplant patients.Methods:A quality enzyme linked immunosorbent assay technique was applied for detection of anti-human PV-B19 IgM (DRG-Germany) and IgG (DRG- Germany) in sera of 50 renal transplant recipients followed up for three successive months and 50 normal were collected from the Center of Kidney Diseases and Transplantation in the Medical City of Baghdad and Al-Khayal Hospital and other many private laboratories, Iraq, during the period from August 2015 till February 2016. The age was ranged from 11-57 years.Results:This prospective study included fifty renal transplant recipients with fifty subjects as a control group. The mean ± standard deviation of ages for RTRs was 32.90 ±12.76, which was comparable to that of healthy controls mean 34.02 ± 14.23 ranging for both RTRs and control group between 11-57 years old. Thirty five (70%) RTR aged less than 40 years, and the remaining fifteen (30%) were above 40 years old, and 38/50 (76%) of these RTRs were males. Seropositivity rate in control was 2/50 (4%) but seropositivity rate in patients was 10% (5 cases) and 10% (5 cases) as turning positive for an individual occurrence of human PV-B19 IgM alone at P value = 0.0299 and 9/50 (18%) from control positive for IgG, while 60% (30 cases) for a similar individual rate of positive human parvovirus B19 IgG at P value < 0.0001.Conclusion:The human PV-B19 infection rate was significantly higher among renal transplant recipients than normal controls. Infections correlated with abnormal renal functions tests, and which may in turn cause anemia and/or reticulocytopenia.Keywords: Parvovirus-B19, anemia, reticulocytopenia, renal transplant recipients, ELISA


Article
6.FREQUENCY OF HEPATITIS C VIRUS GENOTYPES /SUBTYPES ASSOCIATION WITH RESPONSE TO THERAPY IN A SAMPLE OF HCV INFECTED IRAQI PATIENTS

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Background:Hepatitis C virus (HCV) is an important human pathogen affecting 120-170 million individuals in the world. Identification of the causative virus genotype is of a significance to both clinical practices and predict the likelihood to therapy response.Objective:To determine the distribution of HCV genotypes/subtypes and its association with response to therapy among newly diagnosed HCV patients.Methods:Fifty patients with confirmed anti-HCV antibodies were included in this study for HCV genotyping in association with response to therapy. Blood samples from patients were subjected to RNA extraction and reverse transcription step; viral load of HCV was measured by polymerase chain reaction (PCR) at time zero, 3 months and 6 months of dual therapy. Response to therapy was measured as a decrease in viral load (2 log or more) and was described as: good (median log is zero after 6 months of therapy), moderate (median log declines more than 2 log but not zero after 6 months of therapy), poor (median log does not decline or decline less than 2 log after 6 months of therapy).Results:Two genotypes of HCV were detected, genotype 4 was the predominant (27/50, 54%) followed by genotype 1 (23/50, 46%). For HCV subtypes, subtype 1a was of highest percentage (28%) followed by 4e (24%), 1b (18%), 4a (14%), 4b (12%), and 4e (4%).The results revealed a significant association between HCV subtypes, but not genotypes, with response to therapy. HCV subtype 1a followed by 4a showed the highest rate of response 85.7% and 71.4%, respectively, while interestingly HCV subtype 4d showed no response and 1b showed poor response 11.11%.Conclusion:HCV subtypes of great importance in predicting success to HCV therapy and it is believed this would affect the newly emerging directly acting drugs as well.Keywords:HCV, genotypes, response to therapyCitation:Abdulhassan LJ, Abdulamir AS, Alkhalidi NM, Alwaysi SA. Frequency of hepatitis C virus genotypes /subtypes association with response to therapy in a sample of HCV infected Iraqi patients. Iraq. Iraqi JMS. 2018; Vol. 16(1): 30-40. doi: 10.22578/IJMS.16.1.6

Keywords

HCV --- genotypes --- response to therapy


Article
9.ASSOCIATION OF HUMAN HERPES VIRUS 6A INFECTION IN ENDOMETRIAL EPITHELIAL CELLS WITH MISCARRIAGE

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Background:The microbial infections are major cause in abortion, of which viruses appear to be the most frequently involved pathogens. Human Herpes Virus-6 (HHV-6) has been implicated in cases of poor pregnancy outcome.Objective:Detection and quantification of HHV-6 viral load along with titer of IgM and IgG in endometrial epithelial cells of women with unexplained miscarriage (cases group) compared to the decidual endometrial tissue in women with full term conception (the control group).Methods:A total of 90 samples, 45 were taken by curettage from miscarriage women at first and second trimesters of pregnancy and 45 were taken from women with full term pregnancy during caesarian operation. HHV-6 DNA was extracted from uterus lining tissue samples, and then it was detected and measured by real time qPCR; in addition, 3 ml of blood were collected from each subject for detection of HHV-6 IgM and IgG antibodies.Results:HHV-6 DNA was detected in uterus lining tissue of 20% of abortion and 22.22% of controls (P value=0.79). The median viral load of HHV-6 per 106 of uterine cells in abortion group was 15, while for control group was 13 (P=0.54). HHV-6 load increased with increased number of previous abortions, and it increased with presence of recurrent abortion history. It showed significant differences in the median of HHV-6 DNA load between recurrent abortion and non-recurrent abortion groups (P value =0.033). Positive anti-HHV-6 IgM serum antibodies in HHV-6 DNA detected and no-detected subjects were 0%, and 9.85% respectively; the positive anti-HHV-6 IgG serum antibodies in detected HHV-6 DNA were 100%, while 83% in HHV-6 DNA non-detected subjects. The combination of positive/negative results of the anti-HHV-6 IgG and IgM serum antibodies in HHV-6 DNA detected versus non-detected group showed 100% of HHV-6 DNA detected were positive for IgG and negative for IgM, versus 73% in non-detected group. Conclusion:HHV-6 might associated with unexplained miscarriage. HHV-6 infection might increase with increased number of previous abortions, and increased with presence of recurrent abortion history.Keywords:Human Herpes Virus 6A, MiscarriageCitation;Dosh HS, Abdulamir AS, Abdul-Razzaq SH. Association of Human Herpes Virus 6A infection in endometrial epithelial cells with miscarriage. Iraqi JMS. 2019; 17(1): 57-65. doi: 10.22578/IJMS.17.1.9

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