Table of content

Iraqi Journal of Biotechnology

المجلة العراقية للتقانات الحياتية

ISSN: 18154794
Publisher: Baghdad University
Faculty: Institute of genetic Engineering and Techno-biology
Language: English

This journal is Open Access

About

Iraqi Journal of Biotechnology was founded in 2001 ,it was first issued in 2002,it is a semi-annual refereed scientific journal issued by the Institute of Genetic Engineering and Biotechnology in Baghdad University in fields of biology, environment, agricultural sciences ,medicine, dentistry, pharmacology, veterinary medicine and researches specialized in bioinformatics

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E-mail:journal@ige.uobaghdad.edu.iq
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Baghdad-AL-Jadriyah -p.o.box:12074

Table of content: 2012 volume:11 issue:2

Article
PRODUCTION AND PURIFICATION OF PLASMID VECTOR PUB110 IN TRANSFORMED BACILLUS SUBTILIS BACTERIA
إنتاج وتنقية ناقل الإستنسال pUB110 في بكتريا Bacillus subtilis المتحولة

Authors: Khlood A. Al- Khafaji
Pages: 141-150
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Ninety percent of B. subtilis protoplast could obtained after 30 minute from treatment of the bacteria with both Ethylene diamine tetra acetic acid (EDTA) and lysozyme enzyme. Protoplast transformed with the plasmid vector pUB110 had efficiency 104 cell/mg of DNA . Kanamycin resistant cell was gained after culturing transformed bacteria onto regeneration medium which containing kanamycin antibiotic. Three different methods were used for DNA extraction from transformed B. subtilis and the plasmid vector pUB110 could detected from the three methods. Salting out procedure gave the highest DNA yield with chromosomal DNA smear in comparison with the other two procedures. Furthermore, gel electrophoresis for DNA extracted by CTAB method showed high purity DNA extract in comparison with other extraction methods.


Article
DETERMINING THE CELLULASE GENE LOCATION OF SOME PSEUDOMONAS SPECIES ISOLATED FROM SOIL AND STUDYING THE POSSIBILITY TO INCREASE CELLULASE DEGRADATION BY PLASMID DNA AMPLIFICATION TECHNIQUE
تعيين مواقع المورثات المحللة للسليلوز في بعض الأنواع العائدة لجنس Pseudomonas ودراسة إمكانية زيادة قدرة التحلل بإستعمال تقنية تضخيم محتوى الـDNA البلازميدي

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Twenty six cellulose-degrading bacterial isolates were isolated from different soil samples from Nineveh governorate, these isolates were checked for antibiotic resistance towards Tetracycline, Nalidixic acid, Cefalexin, Ampicillin and Chloramphenicol, according to their resistance pattern eight of them were chosen for diagnosis. The study found that all these isolates belonged to Pseudomonas putida. Curing by elevated temperature was carried out to identify the cellulose degradation gene location, curing percentage of some isolates were between (4-31%) which may give a fact that these isolates may carry cellulose genes on their plasmids. On the other hand some isolates failed to be cured probably because their cellulose genes were chromosomal. Plasmid amplification by 150µg/ml chloramphenicol was studied for all isolates that carried plasmid for cellulose degradation, we found that the isolate (19) had the ability to amplify its plasmid content. Certification that cellulase plasmids were amplified was done by comparing the CO2 evolution for the original and the amplified isolate per week.


Article
USING ELISA TECHNIQUE IN DETECTION OF MOLD CONTAMINATION OF TOMATO PAST
إستعمال تقنيه الإيلايزا في كشف التلوث الفطري لمعجون الطماطة

Authors: Abdul Ghani I. Yahya
Pages: 163-174
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Abstract

Indirect ELISA was used for determining antibodies titer in antisera using Anti – IgG – HRP (Goat Anti – IgG-Horse Radish proxidase), the titer of these anti bodies for Aspergillus flavus, Rhizopus stolonifer were 1/160, 1/80. Conjugate was prepared by labeled antisera against A. flavus , R. stolonifer (After precipitation by ammonium sulfate) with alkaline phosphatase, titration of conjugate to each one was carried out using ELISA test, results showed that the titer of A. flavus conjugate was 1:10 and R. stolonifer conjugate was 1:5 The sandwich ELISA test was used for detecting several concentrations of A. flavus, R. stolonifer and their extracts in carbonate buffer solution pH 9.5 involved: 25, 12.5, 6.25, 3.125, 1.56, 0.78, 0.39 micrograms per milliliter. ELISA test also used in detecting their concentration: 1, 2, 3 micrograms of these two fungi per milliliter of tomato paste diluted 1:5 diluted with PSB solution results showed that the absorbance of these concentration in carbonate buffer was more than in tomato paste and positive correlation was found between concentration of mold and the absorbance , ELISA test was compared with biological method (plating culture) for detecting the three concentrations of mold in tomato paste, results showed that ELISA test was more sensitive than plating culture which detect viable mold spores only.


Article
EFFECT OF RADIATION ON NUMBER AND WEIGHT OF SHOOTS IN VITRO FOR
تأثير أشعة كاما في عدد البراعم ووزن النمو الخضري لنبات الموز صنف (كراندناين) المكثر بإستعمال تقنية الزراعة النسيجية

Authors: Abdulkareem R. Kadhim
Pages: 175-181
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Abstract

The aim of this experiment was to study the effect of gamma rays ( 0, 10, 20, 30, 40 and 50 Gy) and time intervals (2and 4weeks) on number and weight of shoots of Grandnaine banana variety propagated in vitro. The result showed that the high doses (20, 30, 40,50 Gy) of gamma ray had significant negative effect on number and weight of shoots while 10 Gy dose showed that the highest number and weight of shoots compared with the control. Moreover significant difference were found in the number and weight of shoots after 4 weeks compared with 2 weeks. The interaction between radiation and time interval showed that the doses 0, 10 and 20 Gy led to a significant increase in number and weight of shoots in 4 weeks compared with 2 weeks, however there were no significant effect between time intervals and the high doses 30, 40,and 50 Gy.


Article
تحديد العوامل الوراثية المسؤولة عن صفة المقاومة للمضادات الحيوية في جرثومة STAPHYLOCOCCUS SCIURI
INVESTIGATING OF THE GENETIC FACTORS RESPONSIBLE FOR ANTIBIOTICS RESISTANCE IN Staphylococcus Sciuri

Authors: Fatima A. Ali
Pages: 182-195
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Abstract

For investigating the genetic factors responsible for antibiotics resistance for Staph. sciuri transformation experiments were carried out with standard strain Escherichia coli JM83 using DNA plasmid purified from 5 strains of Staph. scriui posses predominant resistance differences between (6-24) antibiotics. The mechanism responsible for antibiotic resistance was tested to produc Extend Spectrum Beta lactamase (ESBLs) after using electrophoresis technique using Polyacrylamide gel with sodium dodocyl sulfate (SDS – PAGE) for BLs extracted from the 5 strains produced it and it revealed (1-3) bands with m.wt. between (20-26) KDa...As regard responsible for It was found that this genes are located on plasmid DNA molecules and these results were supported by electrophoresis of DNA purified from plasmid transformed colonies of laboratory strains of E. coli JM83, a single band of plasmid DNA on agarose gel appeared to have between (1-4) bands according to the types of the strains. On the other hand, one strain couldn't give any band which may indicate the occurrence of gene code for antibiotic resistance may be carried by the chromosome too.


Article
EFFECT OF DIFFERENT CONCENTRATIONS OF BA AND SUCROSE ON CALLUS INDUCTION AND INITIATION OF ASEXUAL EMBRYOS IN ORANGE (CITRUS SINENSIS L. OSBECK) NUCELLAR SEED TISSUES IN VITRO
تاثير تراكيز مختلفة من BA والسكروز في إستحثاث الكالس ونشوء الأجنة اللاجنسية من نسيج الجويزاء لبذور أشجار البرتقال (sinensis L. Citrus)

Authors: Kadhim M. Ibrahim
Pages: 196-208
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Abstract

Plant tissue culture techniques were exploited for the micropropagation of sweet orange (Citrus sinensis L. Osbeck) seed nucellar tissues as a source for raising asexual embryos and thereafter plants. Callus was induced from the nucellar tissues and subsequent induction of somatic embryos. Callus fresh weight, number of asexual embryos, shoots and their length and root number were investigated. Sucrose concentrations (2, 4, and 6 g/L) and Benzyl adenine (BA) at 0.0, 0.5, 1.0 and 2.0% were added to Murashiege and Skoog (MS) medium.The addition of 0.5 mg/l of BA in the presence of 2% sucrose to the medium caused highest callus fresh weight (295mg), while supplementation with 1.0 or 2.0 mg/l BA in the presence of 4% sucrose resulted in highest number of somatic embryos per exiced tissue from single seed seed. Supplementation of the medium with 1.0 or 2.0 mg/l of BA and 6% sucrose recorded the highest number of shoots per single tissues. Shoots rasied from callus tissues were rooted on the same medium but containing 0.0 or 0.5 mg/l BA in the presence of 4% sucrose achieving the highest mean number reached 2.5 roots/shoot. Successful acclimatization of plantlets was achieved when they transferred to a mixture of peat moss and riversand media. Acclamatizsed plantlets were transferred successfully to pots with a survival percentage of 80%.


Article
STUDY THE GENOTOXICITY EFFECT OF HOECHST 33258 STAIN IN THE HUMAN LYMPHOCYTE
دراسة التأثير السمي الوراثي لصبغة هوكست Hoechst 33258 في الخلايا

Authors: Muhammad A. Al-A’adhmi
Pages: 209-218
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Abstract

Most stains using in DNA labeling has the ability to cause genetic mutations because of its ability to intercalated on DNA. Hoechst 33258 stain is one of a widely used stains in laboratory preparations, some studies indicated that this stain caused genetic mutations. It is important to detect the relationship between this stain and the people who works in laboratories, there is a lack of studies describing accurately the genotoxicity of this stain. This study is an investigation on the genotoxicity of Hoechst 33258 stain in human lymphocytes (in vitro) using cytogenetic tests including Mitotic Index (MI), Chromosome Aberration (CA), Sister chromatid exchange (SCE) and Preparation of micronuclei, using different concentration of Hoechst stain 5μg/ml, 10μg/ml, 15μg/ml, 20μg/ml. The results showed significant increase in Mitotic Index (MI), Chromosomal Aberration (CA), Sister chromatid exchange (SCE) and Micronuclei this increase is direct proportion with the increase of stain concentration. The Chromosome Aberration represented by chromosomal and chromatid breaks showed at (5,10,15,20)μg/ml, chromosomal and chromatid gap showed at (5,10,15,20) μg /ml, while ring chromosome and dicentric chromosome showed at (15,20) μg /ml.


Article
SOME IMMUNOLOGICAL VARIABLES IN THE BLOOD OF WOMEN WITH TOXOPLASMOSIS IN RAMADI CITY
بعض المتغيرات المناعية في دم النساء المصابات بداء المقوسات في مدينة الرمادي

Authors: Marwa Sh. Dh.Al-Rawi
Pages: 219-226
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The aim of the present study was to evaluate some immunological variables in women with Toxoplasmosis in Ramadi city. A total of 136 blood samples collected from women who were suspected of having Toxoplasmosis in addition to 40 collected Samples blood from apparently-healthy women as controls. Different immunological tests were done including Latex Agglutination Test (LAT) for Toxoplasmosis, Anti-Toxoplasma IgM and IgG by ELISA, and Complement C3 and C4 and IgA by Single Radial Immunodiffusion Assay. It was found that (20.2%) were positive for Anti-Toxoplasma IgM by ELISA and (26.5%) were positive for Anti-Toxoplasma IgG. Levels of IgA were found to be significantly higher in infected patients than controls and levels of complement C3 were also significantly higher in patients than controls while no significant differences were noticed in C4 levels.


Article
TRANSFER OF RI T-DNA GENES OF AGROBACTERIUM RHIZOGENES R1601 VIA DIRECT MICROINJECTION AND CO_CULTIVATION TO CARROT, DAUCUS CAROTA L. TISSUE AND PRODUCTION OF TRANSFORMED HAIRY ROOT CULTURES
إنتقال جيناتT-DNA المحمولة على بلازميدات Ri من بكتريا Agrobacterium rhizogenes R1601 بوساطة الحقن المباشر والزراعة المرافقة الى أنسجة الجزر Daucus carota L. وتكوين مزارع الجذور الشعرية المحورة وراثياً

Authors: Mozahim K. Al-Mallah
Pages: 227-239
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The present study detected the optimal requirements to produce hairy roots on stem segments and root discs of carrot, Daucus carota L., by the natural vector for genetic transformation A.rhizogenes R1601. Direct inoculation of stems segment by bacterial inoculum of density 96x108cell/ml was efficient and sustained the initiation of hairy roots after 10-15 days. They developed on inoculated and non-inoculated sites at percent reached up to 49.9% with a mean of 5.6 hairy roots /segment. Hairy roots excised were as single or clusters of roots and placed on agar-solidified MS medium. They were transferred to MS medium supplemented with gradual concentrations 100, 200, 300 mg/L of antibiotic Cefotaxime through several subsequent transfer to eliminate bacteria. Finally, cultures of hairy roots free from bacteria was developed on MS medium which considered as selection medium. This phenotype of hairy roots was fast growing, brown in color, lacking of root hairs and negatively geotropism. Also, hairy roots were formed on carrot discs, via Co_cultivation with A. rhizogenes R1601 for 8 hours which appeared the best time, after 25 days at a percent of 17.8% and a ratio of 3 hairy root/disc. This phenotype of hairy root was slow in growth, white in colour and express less branching. Paper electrophoresis results demonstrated the present of agropine in both phenotype of hairy roots which proved the incidence of genetic transformation of these roots compared with the normal root (control).


Article
EFFECT OF ULTRA-VIOLET RADIATION ON BIOLOGICAL PERFORMANCE OF THE FIGMOTH,
تأثير الأشعة فوق البنفسجية على الإداء الحياتي لحشرة عثة التين

Authors: Maysoon A. Shawkit
Pages: 240-247
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Abstract

The effect of ultra-violet rays 312 nm wavelength on different egg ages (1,2and3) days of the fig moth, Ephestia cautella as well as the effects on early and late pupa exposed for 3, 6, 9and 15 minutes were studied. Results showed that percent of egg hatching and adult emergence were reduced gradually with increasing exposure period, this is because of higher sensitivity of the embryonic cells in the eggs when exposed to this type of radiation. This study also showed significant differences in many biological parameters for the pupae aged 2 days when compared with 7 days old ,the younger one was more effected, therefore, resulted in increasing pupal and adult malformation .This investigation concluded that uv-radiation can be apply as safe and effective method to control the storage pests.


Article
MOLECULAR CHARACTERIZATION OF ANTIGENS EXTRACTED FROM HYDATID CYSTS OF HUMAN AND OTHER INTERMEDIATE HOSTS

Authors: Ihsan E. Al-Saimary
Pages: 248-260
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Abstract

This research includes a study of Hydatid Cysts E. granulosus (larval stage on the molecular level, where 7 samples of hydatid cysts were collected from parasite intermediate hosts " Human (Liver, Spleen, Lung) and liver of Sheep, Goat, Cattle and Buffaloes". DNA was extracted from germinal layer cells of hydatid cysts which were isolated shortly or preserved for various periods in 70% ethanol . Genetic analysis of isolated DNA from hydatid cysts collected from human and animals was done by Polymerase Chain Reaction (delete) (PCR) to determine genetic variation depending on Random Amplified Polymorphic DNA . In the present study 10 primers have been used, during which the genetic variations were revealed among isolated (extracted DNA) of hydatid cysts which was collected from human and other intermediate hoists except Cows and Buffaloes. The current results of this study have shown the following :1 - It was found one primer (OPA – 01) was able to diagnose sample numbered 1 which represent the isolated DNA of liver hydatid cyst which was obtained from human at age group 10 – 20 years old. 2 - The ability of primer OPC – 10 to determine fingerprinting of DNA sample of Sheep liver hydatid cyst. 3 - The ability of primer OPC – 05 to determine fingerprinting of DNA sample of human spleen hydatid cyst which was obtained from human at age group 30 – 40 years old. 4 - The ability of primer OPE – 07 to determine fingerprinting of DNA sample of Goat liver hydatid cyst. 5– Amplification process to the DNA samples which extracted from Cows and Buffaloes liver hydatid cysts wasn’t completed by using all 10 primers.

Keywords

E. granulosus --- Molecular --- Antigens --- Human.


Article
EFFECT OF PHYSICAL AND CHEMICAL MUTAGENESIS ON THE ABILITY OF LOCALLY ISOLATED BACILLUS STEAROTHERMOPHILUS
تأثير المطفرات الفيزيائية والكيميائية على قابلية بكتريا BACILLUS STEAROTHERMOPHILUS في إنتاج البروتييز

Authors: Asmaa A. Hussein
Pages: 261-269
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Ninety three soil samples were collected from different locations in Basra governorate. From these samples, 53 bacterial isolates of a thermophilic Bacillus spp. were obtained, all these isolates were identified according to their morphological and cultural characteristics and biochemical tests. Results showed that 6 of these isolates are identified as Bacillus stearothermophilus. These isolates were screened according to their ability of protease production, and it was found that all these isolates were protease producers, among them B. stearothermophilus B17 was the most efficient in protease production. Specific activity of protease in crude filtrate of this isolate was 36.4 U/mg protein. Ability of B17 in protease production developed by mutagenesis using physical and chemical mutagens. Results showed that physical mutagenesis using UV irradiation caused to obtain two over producer mutants out of 45 (13.3%) increased in their ability in protease production. Specific activity of protease in crude filtrate of these two mutants are 54.4 and 80.08 U/mg protein respectively in comparison with 36.4 U/mg protein for the wild-type, while chemical mutagenesis by using MNNG was also lead to obtain one over producer mutant out of thirty two (3.1%) increased in its ability in protease production. Specific activity of protease in culture filtrate of this mutant was 99 U/mg protein in comparison with 36.4 U/mg protein for the wild-type.


Article
INDUCTION OF GENETIC VARIATION FOR DROUGHT TOLERANCE IN TWO RICE CULTIVARS AMBER 33 AND AMBER BAGHDAD
إستحداث تغايرات وراثية لتحمل الجفاف في صنفي الرز عنبر 33 وعنبر بغداد

Authors: Asmaa K. Aurabi
Pages: 270-281
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Seeds of two rice cultivars Amber 33 and Amber Baghdad were presoaked in the chemical mutagen Sodium azide at the concentrations 0.0, 0.5, 1.0 or 2.0 mM for 2, 4 or 6 hrs. The effect of Sodium Azid was examined on seed germination, shoot and root length and promising dose. To increase the genetic variation for drought tolerance, seeds treated with the optimum dose that made 40% in growth reduction in seedling height. Calli were induced from mature embryos on appropriate medium and then transferred to a medium containing 0.0, 0.5, 1.0, 1.5 or 2.0% of Polyethylene Glycol (PEG) (W/ V). Differences were recorded between cultivars and treatments with respect to the studied traits. Results showed that there were no significant differences between cultivars in respect to seeds germination shoot and root length while these parameters decreased with increasing mutagen concentration and soaking time. Results also revealed that there were significant differences between cultivars in % of callus induction and callus fresh weight when callus cultures were transferred to different combinations of 2,4-D and Kin, while there were no significant differences in callus fresh and dry weights between cultivars with increasing concentrations of Sodium Azide and PEG. Callus fresh weight decreased with increasing PEG level in the medium for seeds not exposed to Sodium Azide.


Article
IDENTIFICATION OF PSEUDOMONAS AERUGINOSA FROM BURN WOUNDS ISOLATES BY PCR USING EXOTOXIN A-SPECIFIC PRIMERS.
تشخيص بكتيريا Pseudomonas aeruginosaالمعزولة من جروح الحروق بواسطة تفاعل البوليمريز المتسلسل بإستعمال بادئات خاصة بالذيفان الخارجي أ.

Authors: Iman A. Hussien
Pages: 282-291
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The purpose of this research was to evaluate the use of PCR for the identification of burn wounds isolates of Pseudomonas aeruginosa by amplifying a 396-bp region of the exotoxin A (ETA) structural gene sequence. Specific primers amplified ETA-positive P. aeruginosa DNA, whereas other species of Pseudomonas and bacteria other than Pseudomonas which were isolated from burn wounds infections did not yield any 396-bp fragment. The specificity and sensitivity of the assay were 100 and 96%, respectively, which confirms the assay's reliability for diagnostic and epidemiological studies. The assay can detect 0.2 pg of P. aeruginosa DNA per reaction mixture (3ul) by ethidium bromide staining of an agarose gel. This method is rapid and less cumbersome than other diagnostic methods for the identification of P. aeruginosa strains. The method described can be used to detect a low level of P. aeruginosa from clinical samples without the use of selective media or additional biochemical tests.

Keywords

burn wound --- P. aeruginosa --- ExotoxinA --- PCR.


Article
MOLECULAR VARIATIONS OF MAIZE CMS POPULATIONS AND SUBPOPULATIONS
التغايرات الجزيئية لمجتمعات أصلية ومشتقة لسلالات عقيمة من الذرة الصفراء

Authors: Ayoub O. Alfalahi
Pages: 292 -312
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RAPD DNA markers were used to evaluate trends in genetic diversity among 20 of 24 cms line populations derived from early and advanced cycles of selection. All of the 10 RAPD primers used for initial screening were found to be polymorphic. A total of 139 DNA fragments were generated by 10 random decamer primers, with an average of 13.9 easily scorable fragments per primer. The number of amplified fragments produced per primer ranged from 6 for OPC-02 primer to 21 for OPC-07 and OPE-07 primers, with molecular size ranged from 264bp to 2717bp. The total number of polymorphic fragments and the percentage of polymorphism were 109 and 78%, respectively. Maximum level of polymorphisms were (94%) and (93%) observed for the primers OPD-05 and OPC-08, respectively. Primers OPC-02 and OPD-08 showed the lowest percentage of polymorphism, which were about (50%) and (53%), respectively. Based on the bivariate (1-0) data and genetic similarity with the use of UPGMA cluster method, the dendrogram separated the studied populations into five major clusters I, II, III, IV and V. Cluster analysis which compared between original and subpopulations places in the dendrogram, showed that selection plays an important role in developing new populations via selfing. R6O and its progeny R6S showed a diverged genetic background against other populations. Genetic similarities, computed by Nei and Li’s similarity coefficient revealed that the highest estimate (0.97) was observed between the original and subpopulation of R6. Meanwhile the lowest genetic similarity (0.76) was detected between the original and subpopulation of R2. The highest genetic similarity among the different populations was 0.89 observed between R3O and both R4 populations, whereas genetic distance widened slightly in R6 population after three cycles of selection as R6s possessed the lowest GS value (0.57) against A5O. Relationship between genetic diversity and hybrid vigor was fair enough. Results indicated that DNA molecular markers were highly efficient in detecting the purity and genetic relationship among maize cms populations.

Keywords

Maize --- CMS --- RAPD Markers --- Genetic Similarity


Article
MUTAGENIC EFFECTS OF FLUORINE GIVEN BY DRINKING WATER ON THE CHROMOSOMAL ABERRATION AND MICRONUCLEI IN WHITE MICE MALE
التغيرات الوراثية لفلوريد الصوديوم (NaF) المعطى عن طريق ماء الشرب وشذوذ كروموسومات ونويات ذكور الفئران البيضاء

Authors: Bassim M. Jawad
Pages: 313-320
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The present experiment was designed to study the mutagenic effects of fluorine given by using NaF in drinking water on the chromosomal aberration (CA) and Micronuclei (Mn) in white mice male for three month. (65) male mice divided into three groups the 1st and 2nd groups daily intake (150 and 300 ) mg/L NaF indirectly by drinking water give NaF, while give tap water to the control group. The cytogenetic analysis showed that NaF was causes significant increased (P≤0.01) in M.n and C.A which include “break, ring, delayed and gap” so concluded that fluorine toxicity has dose related effects.


Article
SEQUENCES AND EXPRESSION OF THE ACTIVE LYSOSTAPHIN GENE FROM STAPHYLOCOCCUS SIMULANS ISOLATED OF BOVINE MASTITIS AND ITS BACTERICIDAL EFFECT ON Staphylococcus Aureus
تحديد تسلسل وتعبير جين اللايسوستافين من

Authors: Jalal Y. Mustafa
Pages: 321-339
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Sixty milk samples were recovered from cows suffering from clinical mastitis in Basra. Among these 55 (91.66%) gave good growth in blood agar. After microscopical and biochemical tests, 49 isolates have been diagnosed as G+ bacteria staphylococci. Thirty (50%) isolates of S. aureus are coagulase positive staphylococci (CoPS) and 19 (31.66%) isolates were coagulase negative staphylococci (CoNS). Among the recovered isolates, two isolates are S. simulans and only one gave good antimicrobial activity against S. aureus on the basis of inhibition zone. The active lysostaphin gene with determined size and sequences are 738 bp was isolated and then cloned to E. coli. A lysostaphin gene was prepared with terminal Histidine group in order to isolate and purify lysostaphin protein by used special primers. His-tag column was chosen for isolation and purification of protein with in short time. A high fidelity in running, one band of protein in polyacrylamide gel was seen in comparison with standard protein from Sigma which gave more than two bands, after running in polyacrylamide gel, the molecular weight was about 27 KDa. The antibacterial effect of lysostaphin against S. aureus was studied in Vitro which gave good inhibition zone on sold media. The LD50 of lysostaphin was determined and there were no effects of lysostaphin on mice with all concentrations used. The effect of lysostaphin against S. aureus in Vivo was studied by inducible infection in mammary glands (mastitis). The antibacterial activity of lysostaphin showed significant effect. Finally the histological studies of mammary glands showed the significant activities of lysostaphin to inhibit the growth of S. aureus. To eliminate the antibiotics resistant building this study is conducted. The aims of the study: Isolation and identification of Staphylococcus simulans as a source of lysostaphin producer from mastitis cows. Cloning the lysostaphin gene, using expression of this gene as bactericidal effect on S. aureus in plate. Screening the effect of lysostaphin on induced infected mice (locally) with S. aureus.


Article
DETECTION OF GENE EXPRESSION OF SERINE PALMITOYLTRANSFERASE (SPT2) IN MOUSE CELL LINE RAW264.7 INFECTED WITH LEISHMANIA MEXICANA AMASTIGOTES
التحري عن التعبير الجيني لإنزيم (SPT2)

Authors: Hayder Z. Ali
Pages: 340-348
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Leishmania species are the causative agent of a tropical disease known as leishmaniasis. Previous studies on the old world species Leishmania major, showed that the amastigotes form which resides inside the macrophage of the vertebrate host, utilize host’s sphingolipids for survival and proliferation. In this study, gene expression of serine palmitoyltransferase (SPT) subunit two (MmLCB2) of the mouse macrophage cell line (RAW264.7), which is the first enzyme in the de novo sphingolipid biosynthesis, was detected in both infected and non-infected macrophages. This was detected under condition where available sphingolipid was reduced, with the new world species Leishmania mexicana. Results of qPCR analysis showed that there was no difference in the expression of MmLCB2 in infected and non-infected macrophages, under normal and serum-reduced media, suggesting that host sphingolipid did not up-regulated during infection. This can be concluded as a difference between the Old and New world Leishmania on the level of host-parasite interaction.

Keywords

Leishmania mexicana --- SPT --- RAW264.7 --- myriocin.


Article
GENETIC CHANGES IN LOCAL METHICILIN RESISTANT Staphylococcus aureus LEADING TO HETRO-VANCOMYCIN SUSCEPTIBILITY

Authors: Dlnya A. Mohammed
Pages: 349-356
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Methicillin-resistant Staphylococcus aureus (MRSA), which emerged as a major nosocomial pathogen, is now spreading rapidly worldwide. Moreover, treatment of S. aureus infections is becoming more complicated. The emergence of glycopeptide resistance in S. aureus is considered to be a serious threat around the world, since current treatment of serious infections caused by MRSA relies mainly upon the administration of glycopeptide antibiotics. To understand the mechanism of vancomycin resistance, resulting from multiple mutations, in clinically isolated S. aureus. Complete sequences of multiple genes, including two-component vancomycin histidine kinase sensors (VraS), vancomycin response regulator (VraR), teacoplanin resistant gene (trf AB ) were compared with those of their susceptible strain. Further genetic analysis was performed on 9 vancomycin- resistance S. aureus (VRSA) revealed that a single point mutations leading to amino acid substitutions in a gene: vraS, encoding a two-component histidine kinase sensor. The accumulation of the mutation in proteins VraR regulator, correlated with further increases in the glycopeptide MIC. Genetic analysis of 9 VRSA isolates also identified the predominant amino acid substitutions in VraS: 5 isolates revealed mutations in VraS gene, followed by 3 isolates with mutations in Vra R Finally the present research provides novel insights into genetic diversity of VraSR mutants among clinical S. aureus isolates with variable susceptibility to vancomycin.


Article
DETECTION OF A, A- MUTATIONS OF G6PD GENE ON MOLECULAR LEVEL IN IRAQI POPULATION
الكشف عن طفرات A, A- المصاحبة لأنزيم نازع هيدروجين الكلوكوز-6- فوسفات (G6PD) على المستوى الجزيئي بإستعمال تقنية PCR/RFLP في عينات محلية

Authors: Rana A. Al-Temmemy
Pages: 357-368
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The study involved the extraction of deoxyribonucleic acid (DNA) from 71 samples of random G6PD patients and 85 samples apparently healthy individuals from different Iraqi populations respectively, which was then amplified by polymerase chain reaction (PCR) and later subjected to digestion by restriction enzymes (Nla III and Fok I) to create restriction fragment length polymorphism (RFLP) to enable the detection of mutation that caused G6PD deficiency namely A and A-. The results of the current study showed that Iraqis were affected by G6PD deficiency in a percentage 7.2% and showed that the affected cases were attributed to A mutation in 4.2% while 0% was recorded for A- mutation.


Article
ADHERENCE OF TYPE 1 FIMBRIATED ESCHERICHIA COLI TO UROEPITHELIAL CELLS OF WOMEN WITH DIABETES MELLITUS
التصاق بكتريا القولون Escherichia coli المهدبة بخمل النمط-1 بالخلايا الطلائية البولية لنساء مصابات بمرض السكري

Authors: Rita N. Rammo
Pages: 369-376
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Adherence of type 1 fimbriated Echerichia coli to uroepithelial cells of diabetic and non-diabetic women as well as the inhibition of adherence by the presence of anti-type 1 fimbriae at different dilutions have been studied. One hundred thirty nine isolates were collected from different clinical sources. Ninety isolates were characterized as E. coli by morphological, microscopic and biochemical tests. Sixty one % of the E. coli isolates were identified as being type 1 fimbriated, mannose sensitive hemagglutination (MSHA) and the rest (39%) as being non-type 1 fimbriated, mannose resistant hemaggllutination (MRHA). The number of type 1 fimbriated E. coli adhered to uroepithelial cells isolated from diabetic women (41.04 ± 2.43) was about twice that of control (16.48 ± 2.3). Partially purified type 1 fimbriated E. coli was mixed with Freund's adjuvant (CFA and IFA) and subsequently injected subcutaneously into rabbits via three injections in 2 weeks intervals. Anti-type 1 fimbriae (antibodies ) that had been raised in rabbit blood serum were pre-incubated with type 1 fimbrated E. coli. Inhibition in adherence ranged between 62 to 72 % depending on the dilution of antisera. Moreover, it has been found that low dilution of antisera causes higher inhibition of type 1 fimbriated E. coli to diabetic uroepithelial cells.


Article
DETERMINATION GENETIC FACTORS CONTROLLING Β-LACTEMASE ENZYME RELATED TO BLA TEM AND BLA SHV FAMILIES USING THE POLYMERASE CHAIN REACTION
تحديد العوامل الوراثية المسيطرة على إنتاج أنزيم البيتا لاكتاميز من عائلتي bla TEMوbla SHV بإستعمال التفاعل التضاعفي لسلسلة الدنا

Authors: Sawsan S. Al-Jubori
Pages: 377-388
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Clinical isolates (75) of Gram- negative bacteria were isolated from patients with different infections at Al-Kadhumia Educational hospital in Baghdad. These isolates were diagnosed using different morphological and biochemical test followed by the complementary api 20E and api 20NE test. They were distributed as Escherichia coli, Proteus mirabilis and Pseudomonas aeruginosa (25 isolates for each). β-lactemase production was investigated using the rapid iodometric method and the results revealed that 48 isolates out of 75(64%) gave a positive result .These isolates were, 21 from E.coli (84%),17 isolates for P. mirabilis (68%) and 10 of Ps. aeruginosa(40%).The double disc approximation test was used to detect Extended spectrum β-lactemases (ESBLs)and the result showed that 28 isolates out of 48(58.33%)gave positive result as 19,3,6 isolates for E. coli, P. mirabilis and Ps. aeruginosa respectively. Polymerase chain reaction(PCR) technique was performed to detect the genetic factors controlling β-lactemases related to bla TEM and bla SHV families and template DNA was prepared either by the boiling method in which the total genomic DNA will serve as a template or by using the plasmid DNA as a template and the later was prepared by the alkaline lysis method. Results of using total genomic DNA showed that 27 isolates out of 28(96.42)were harboring bla TEM gene based on the presence of 950 bp bands in 1% agarose gel .By using the plasmid DNA as a template ,only 10 isolates (35.7%) gave clear band which may indicate that β-lactemases of these isolates may be are plasmid mediated .For bla SHV gene , the results of using the first template showed that only 2 E. coli isolates out of 28(4.14%)gave a positive result represented by presence of 800 bp band in the agarose gel while no band was observed when the plasmid template was used which may indicate that the enzymes here under chromosomal control.


Article
IN VITRO REGENERATION AND SOMATIC EMBRYOGENESIS IN CITRUS
الإكثار المعملي والأجنة الجسدية في الموالحالإكثار المعملي والأجنة الجسدية في الموالح

Authors: Adel M. El-Sawy
Pages: 389-406
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Plant regeneration through somatic embryogenesis was investigated in Washington navel orange, Satsuma mandarin, Lemon, Variegated lemon, Lime, Citron, Pummelo, Rough lemon , Sour orange , Volkamer lime and Rangpur lime using stigma, style and ovary as explants. Explants were cultured on Murashige and Skoog( MS )medium supplemented with50 gl-1 sucrose, 500 mgl-1 malt extract and 3 mgl-1 Benzyl adinen (BA). Two months later, somatic embryos were induced from embryogenic callus. The percentage of embryo formation and number of embryo germinated depended on the explants type and genotype. Best results were obtained when stigma explants of variegated lemon and Citron were used. After ten months, somatic embryos developed into plantlets at a frequency ranged from13.3% for Lime to 66.7 % for Lemon. Virus presence was tested by ELISA and RT-PCR . The obtained results indicated that the plantlets regenerated through somatic embryogenesis were CTV-free. RAPD analysis was used to asses the genetic stability of obtained plantlets as compared to the mother plants. Results indicated that most plantlets were belonging to the respective mother plants and the polymorphism percentage was genotype and explant- dependant. Abbreviations: BA – Benzyladenine; CTV – citrus tristiza virus; TCL – Thin cell layer ; CPsV – citrus pserosis virus; GA3 – Gibrillic acid.


Article
DNA PATERNITY TEST AND STATISTICS FOR FATHER-DAUGHTER INCEST CASE
فحص البنوة في قضية وطأ المحارم (أب وإبنته) بتقنية البصمة الوراثية والإحصائيات الخاصة بها

Authors: Ammirah J .Omar
Pages: 407-413
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Incest is sexual intercourse between close relatives (1,2) that is either illegal in the jurisdiction where it takes place or socially taboo. The type of sexual activity and the nature of the relationship between people that constitutes a breach of law or social taboo vary with culture and jurisdiction. Father-daughter was the most commonest form of incest, its prevalence is difficult to assess due to secrecy and privacy (3,4). In Iraqi law incest is a Forensic case usually referred to Medico legal Institute to be investigated with a high degree of care as a disputed paternity due to sever , danger sequel of punishment may reach to perpetrator execution. Aim of this study is to focus the light and report one of the important incest cases referred to Medical-Legal Institute .DNA profiles of 15 autosomal Short Tandem Repeat and Amelogenin markers were investigated for the (Father and father, s cousin, Mother and son) in DNA Typing Department. This case was rebated according to the match of all genetic markers of DNA profile of the son with that of the putative father (Mother s Father ). We conclude that the father can not be excluded from being a biological father of son who has 8 homogenous out of 15 loci suggesting high kinship relatedness between his father & mother. The Liklihood ratio was 99.99 %.


Article
DETECTION OF HOMOZYGOUS G.IVS5+1G>A,
تعقب الطفرات الوراثية متجانسة الزيجة g.IVS5+1G>A, g.IVS34-1G>C, c.886C>T في مرضى تضخم الغدة الدرقية السام و سرطان الغدة الدرقية في العراق

Authors: Abdulhussein M. AL-Faisal
Pages: 414-421
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Forty eight DNA samples from 31 toxic goiter patients and 17 thyroid cancer patients were analyzed to detect g.IVS5+1G>A, g.IVS34-1G>C and c.886C>T mutations of the thyroglobulin (TG) gene. Among these samples three homozygous mutations were detected. Two of these mutations were detected in two toxic goiter patients as guanine to adenine transition g.IVS5+1G>A at position +1 of the donor splice acceptor site in exon-intron 5 and transversion that replaced guanine by cysteine (g.IVS34-1G>C) in the exon 34. The third homozygous mutation was detected among one thyroid cancer patient as transition that replaced cysteine by thymine c.886C>T in the exon 7. No homozygous g.IVS34-1G>C mutation was detected in thyroid disorders before, to our knowledge this is the first time by which homozygous mutation was detected in one toxic goiter patient.


Article
MOLECULAR BIOLOGY OF ENTEROTOXIC GENES PROFILES OF STAPHYLOCOCCUS AUREUS ASSOCIATED WITH SUB-CLINICAL MASTITIS IN DAIRY COWS IN SULAIMANYAH PROVINCE
البايولوجي الجزيئي لنسق جين السم من المكورات العنقودية الذهبية المرتبطة دون السريري التهاب الضرع في الأبقار الحلوب في محافظة السليمانية

Authors: Essam F. Al-Jumaily
Pages: 422-432
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The study were conducted on 250 dairy cows (985 quarters) selected randomly from six selected dairy herds and also from individual cows from areas around the Sulaimani region to investigate the coagulase negative and coagulase positive Staphylococcus species caused clinical and sub-clinical mastitis in dairy cattle. The prevalence of bovine mastitis was 80 % (200 cows). Production of enterotoxins A, B, C and D were detected by a reverse passive latex agglutination test (RPLA). Among 246 coagulase positive staph. (CoPS) isolates, 113 (45.9%) were found to be positive for production of one or more of enterotoxin types A, B and C, while none of the coagulase negative staph. (CoNS) isolates had the ability to produce enterotoxin. The technique of polymerase chain reaction was applied to determine the presence of gene that coded for staphylococcal pantone-valantine lucocidine (luk gene) and staphylococcal enterotoxin (se gene). Among 246 S.aureus and 95 CoNS isolates only 10 isolates(4.1%) were found to carry the luk gene. None of the CoNS isolates carried the (luk gene). And also from 246 Staphylococcus aureus isolates, 125 isolates (45.9%) were found to be involved in production of one or more se genes, out of 125 S. aureus enterotoxin producer, it was found that 66 isolates (52.8 %) produce enterotoxin type A; 27 isolates (21.6%) produce enterotoxin type B; 20 isolates (16 %) produce enterotoxin type C and 12 isolates (9.6 %) produce enterotoxin type E. None of the isolates tested produce enterotoxin type D. The results indicated that S. aureus isolated from bovine mastitis has the ability to produce enterotoxin, while CoNS did not have the ability to produce enterotoxin. Additionally, it was demonstrated that S. aureus causing mastitis in Sulaimanian dairy herds harbored the sea, seb, sec and see genes and produced the SEA, SEB ,SEC and SED toxin, suggesting that it may play a role in bovine mastitis pathogenesis.


Article
DETECTION OF SERUM TUMOR MARKERS CA15-3 AND CEA IN BREAST CANCER PATIENTS
تحديد بعض معلمات الورم في مرضى سرطان الثدي

Authors: Ola H. Al_Bayati
Pages: 433-444
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Breast cancer is the commonest type of cancer affecting women worldwide. Different studies have dealt with the etiological factors of that cancer trying to find a way for early diagnosis and satisfactory therapy. This study is confined to the malignant subtypes invasive ductal carcinoma which, by far, the commonest type of breast cancer and invasive lobular carcinoma, designated to study the relationship between breast cancer patients and their concentrations of serum tumor markers CA15-3, CEA and correlate these concentrations with the stage of disease. The study consisted of 50 patients who were diagnosed as breast cancer patients they have attended to AL-Kadhemya Teaching Hospital (all were females) and 25control group apparently normal individuals with comparable age range of patients. The blood samples (5 ml) of venous blood was collected from all of studied cases in order to be used for immunological study by measuring their serum level of serum tumor markers CA15-3 and CEA using Enzyme-linked Immunosorbent Assay (ELISA) technique. The results have shown significant relation of CA15-3 which elevated in 46% of patients, while the serum tumor marker CEA levels were not significant which increased in 30% of patients.


Article
EVALUATION OF IMMUNOGLOBULIN M (IGM) AND INTERLEUKIN 2 (IL-2) ASSAY IN TRUE INFECTION OF ENTAMOEBA HISTOLYTICA
تقيم مستوى الضد م والمدور المناعي الثاني في الإصابة الحقيقية للزحار الأميبي

Authors: Ghada B. Al-Oumashi
Pages: 445-454
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Eighty six sera samples were collected from patient suffering from gastrointestinal symptoms (less than one year-67 years old) had been established from the beginning of May to the end of November 2010 who attends to the Maternity and Childhood Teaching Hospital and Al-Diwania Education Hospital in Al-Diwania Governorate. Nested polymerase reaction and restriction endouclase were done previously and classified the specimen into positive for Entamoeba histolytica mixed with E. dispar , positive for E. dispar only and negative for Entamoeba sp. and usually control groups. IL-2 was detected by using ELISA in microtiter plate and designed the concentration of it in the serum. Data were translated into a computerized database structure. An expert statistical advice was sought for. Statistical analyses were computer assisted using SPSS version 13. The result showed there was an elevation in the median concentration of IgM and IL-2 in positive group for E. histolytica mixed with E. dispar in comparing with control groups, and those which was negative for Enatmoeba and positive for E.dispre only. In conclusion the levels of the serum. IgM and IL-2 which are a mediator of inflammation gave a high sensitivity and specificity in relationship with invasive amoebasis due to the high titter of IgM and IL-2, so it can be used in early diagnosis of E. histolytica infection.


Article
PUTATIVE GENETICALLY MODIFIED CALLUS DERIVED FROM TRANSFORMED HAIRY ROOTS INDUCED ON SUGARBEET (BETA VULGARIS) EXPLANTS BY AGROBACTERIUM RHIZOGENES 1601 HARBOURING RI-PLASMID
تكوين الكالس المعدل وراثياً المشتق من الجذور الشعرية المحولة وراثياً المتكونة على أوراق البنجر السكري Beta vulgaris ببكتريا Agrobacterium rhizogenes 1601 الحاوية على بلازميدات Ri

Authors: Mzahem K. Al-Mallah
Pages: 455-463
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Hairy roots were induced on leaf explants excised from sugarbeet (Beta vulgaris) plants when inoculated by Agrobacterium rhizogenes R1601. Paper electrophoresis proved the presence of agropine, the unusual amino acid in these roots which demonstrated the transformation of these tissues. Transformed hairy roots were easily grown on agar-solidified Murashige and Skoog, 1962 (MS) medium containing 0.5mgl-1 2,4dichlorophenoxy acetic acid (2,4-D) or B5 medium supplemented with a mixture of 0.7 mgl-1 Benzyl Adenine (BA) and 0.7 mgl-1 (2,4-D). Callus produced from these hairy roots grown on MS medium was of friable and white in color, while brown in color on B5 medium. Calli were grown efficiently in liquid B5 medium provided with a mixture of 1.0 mgl-1 kinetine (Kin.) and 1.0 mgl-1 2,4-D. These results may contribute in transformation of sugarbeet plants.


Article
GENOTYPING OF THE ABO BLOOD GROUP SYSTEM IN IRAQ POPULATION USING PCR-RFLP
التنميط الوراثي لمجموعة الدم ABO للمجتمع االعراقي بإستعمال PCR-RFLP

Authors: Mohammed I. Nader
Pages: 464-474
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Due to a large number of alleles that could give similar phenotypes but differ in genomic structure. This study study was conducted to determine the ABO genotyping by use PCR-RFLP method. Blood samples withdrawn from 30 unrelated individuals who appear to be healthy. ABO system phenotype grouping was detected by two immunoagglutination tests. Slide and ELISA were done for all samples. Homozygous allele was selected by chosen special family which gives known offspring according to Mendel laws. The DNA was extracted by using two methods, the manual method by using extra gene kit gives relatively high yield concentration (5-10µg/ml) but slightly low purity (0.9-3.3) and the automated BioRobot EZ1 gives slightly lower yield of DNA(4-8 µg/ml) but high purity (1.6–2.2) in comparison with manual method . Two separate segments of ABO gene the glycosyltransferases gene (216pb) in exon 6 and (703pb) in exon 7 were amplified using two sets of specific primers. Restriction digisten of the two amplification PCR productes was used for discrimination of AB and O allells.


Article
HEPATITIS C VIRUS GENOTYPES IN IRAQ
النمط الوراثي لفايروس التهاب الكبد نمط C في العراق

Authors: Mohammad D. Khalid
Pages: 475-480
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This study included 210 Hepatitis C Virus(HCV) infected patients 150 males and 60 females with an range of 3-50 years, they are 100 patients from thalassemic HCV infected, 100 patients from chronic liver disease HCV infected, and 10 persons from blood donors HCV infected. The study show 94(94%),85(85%),80(80%)from thalassemic and chronic liver disease and blood donors HCV infected carrier the HCV, genotype (4) respectively, and 5(5%), 10(10%). 20(20%) respectively carrier 1b genotype and 1(1%),5(5%)from thalassemic and chronic liver disease HCV infected respectively carrier mixed genotype (1b and 4). While 2(20%) from blood donor HCV infected HCV carrier 1b genotypes.

Keywords

HCV --- Genotype --- Iraq


Article
THE ROLE OF PLASMIDS IN ANTIBIOTIC SUSCEPTIBILITY OF LOCAL ISOLATES OF Klebsiella pneumoniae
دور البلازميدات في الحساسية الدوائية لبكتريا K. pneumoniae المعزولة محلياً

Authors: Meelad A. Al-Nasiri
Pages: 481-493
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Four local isolates of Klebsiella pneumoniae subsp. pneumoniae from different sources were tested towards twenty six antibiotic disks. The results showed that all selected isolates were sensitive to nalidixic acid, amikacin, and ciprofloxacin and the isolate MK6 showed further sensitivity to tetracycline, kanamycin, and streptomycin; but all isolates showed resistance toward the rest of antibiotics used. The minimal inhibitory concentration (MIC) test of four selected Klebsiella pneumoniae isolates (MK1, MK2, MK6 and MK20) to nine antibiotics showed the highest MIC values for the selected isolates were for amoxicillin and chloramphenicol (≥ 16384 µg/ml), while the lowest MIC was for streptomycin as it ranged from (8 – 16 µg/ml). Agarose gel electrophoresis was performed to detect plasmid profile for the four selected isolates. The results showed that all isolates exhibited two bands of small plasmid DNA with molecular size ranged from less than 4kb for the smaller one to more than 12kb for the larger one as compared with DNA of molecular size marker )lambda phage restricted with EcoRІ and HindIII). Besides two bands of large plasmid DNA were obtained from the isolates MK20, MK1, and MK6, but the isolate MK2 didn’t show any band of large plasmid DNA. This may be due to a technical error or to the low DNA concentration during extraction process In order to determine whether the plasmids of selected isolates play a role in antibiotic sensitivity, plasmid curing experiment was performed by using physical agent (elevated temperature). The results indicated that high temperature (47°C) was efficient to cure all the plasmids, and the isolates MK20, MK1, and MK2 became sensitive to streptomycin, kanamycin, and tetracycline.


Article
VEGF EXPRESSION IN TROPHOBLASTIC TISSUE OF WOMEN WITH SPONTANEOUS MISCARRIAGE AND INFECTED BY TOXOPLASMA GODNII USING INSITU HYBRIDIZATION
التعبير الموقعي لعامل النموVEGF في النسيج المغذي للجنين (التروفوبلاست) لنساء تعرضن للإجهاض الفجائي نتيجة الإصابة بداء المقوسات

Authors: Nidhal A. AL-Mohaimen
Pages: 494-502
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A total of fifty women, aged between (16-42) years, were involved in this study. Enzyme linked immunosorbent assay (ELISA) test was used for the detection of specific IgM (using serum samples) and insitu hybridization method was used for the detection of VEGF m RNA in trophoplastic tissue. Samples were classified into three groups: Group A-patients with spontaneous miscarriage and Toxoplasma godnii positive (n= 20 women), with a mean age of(23.8±1.631);Group B-patients with spontaneous miscarriage and Toxoplasma godnii negative (n=20 women), with a mean age of (25.5± 1.60); Group C- control group, women with induced abortion for medical causes (n=10 women), with a mean age of (26.4± 1.628). The highest percentage of Vascular Endothelial Growth Factor mRNA expression (65.30± 3.587)% was within the induced abortion group, while the lowest expression was within (A) and (B) group (35.45± 2.184)% and (37.65± 2.964)% respectively. These results indicated that VEGF is essential factor in the process of blood vessel formation in the successful pregnancy, and its low level in miscarriage groups indicated its role in the etiology of miscarriage irrespective of toxo status.


Article
A GENETIC STUDY ON ENTEROCIN – PRODUCTION FROM Enterococcus faecalis ISOLATED FROM DIFFERENT CLINICAL SOURCES
دراسة وراثية على الانتروسين المنتج من بكتريا Enterococus faecalis المعزولة من مصادر سريرية مختلفة

Authors: Nuha Joseph Kandela
Pages: 503-518
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This study include genetic investigation Locally isolated Enterococcus. faecalis ability to produce enterocin, Hemolysin and cytolysin (production hemolysin and enterocin). The results of detection virulence factor showed that E.faecalis isolates possessed 14% hemolysin, of which 8% from clinical sources, and 6% of these isolates showed ability to produce cytolysin , 6% of normal flora isolates produced hemolycin was distributed between 4% isolates produce cytolycin and 2% non-produced. The results of plasmid content showed that selected isolates E.faecalis (EFU32 and EFU36) from Urinary Tract Infection and E.faecalis EFUW47 from Wound infection contained large plasmid bands; while other selected isolates E. faecalis EFW46 from Wound infection and normal flora did not exhibit this band. It was also observed that clinical and non-clinical isolates have homogenous plasmid profile in spite of the variation in their ability in producing enterocin, hemolysin and resistance to antibiotics. The results of plasmid curing by using physical and chemical agents showed the physical agents (heating at 48ºC) was more efficient to curied small and large plasmid completely together with the lots of hemolysin and enterocin production ability, and resistance to some antibiotics. the results of the treatment with chemical agents like mitomycin – c and ethidium bromide, showed the abilty of mitomycin – c agent to cure all small and large plasmid bands for all isolates at concentration of 80 µgml, also lose the ability to produce enterocin and hemolycin was observed. while the results showed variety when treated with eithidium bromide. In order to investigate the role of the large plasmid for enterocin production and whether it is a conjugative plasmid or not, an experiment on conjugation was carried out between cured EFU36 which harbor large plasmid as donor cell and cured EFW46 that did not harbor large plasmid, as a recipient cell. The results of conjugation experiment revealed that genes encoding for the production of enterocin, resistant to vancomycin may located on conjugative large plasmid.


Article
DETECTION OF AUTOLYSIN, PNEUMOLYSIN AND PNEUMOCOCCAL SURFACE ADHESION A GENES AMONG STREPTOCOCCUS PNEUMONIAE CAUSING BACTERIAL MENINGITIS IN CHILDREN
التحري عن جينات PNEUMOCOCCAL AUTOLYSIN, PNEUMOLYSIN, SURFACE ADHESION A في بكتريا STREPTOCOCCUS PNEUMONIAE المسببة لإلتهاب السحايا البكتيري في الأطفال

Authors: Huda Z. Al-Banae
Pages: 519-528
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The aim of the present study is to assess the presence of autolysin, pneumolysin and pneumococcal surface adhesion A genes as virulence factors that may contribute in pathogenicity of Streptococcus pneumoniae causing bacterial meningitis in children. Non-culture tests such latex agglutination test and PCR for S.pneumoniae diagnosis should be considered for patients who need early identification of pathogens or have previously received antibiotics, or whose initial cerebrospinal fluid Gramʼs stain is negative with negative culture at(72)hrs incubation. There were (303) cases delivered their cerebrospinal fluids samples to the central public health laboratory distributed by (183) male (60.39%) and (120) female (39.60%). Seventy seven cases (25.41%) were diagnosed as bacterial meningitis; (36) cases (11.88%) as viral meningitis. S. pneumoniae was isolated from (16) cases (20.7%) of bacterial meningitis. DNA from S. pneumoniae was extracted then subjected to amplification by PCR leading to detection of autolysin and pneumolysin genes in (9) out from (16) isolate and presence of pneumococcal surface adhesion A gene in (8) out from (16) isolate.


Article
ASSOCIATION OF CELIAC DISEASE WITH HLA-DRB1 AND HLA-DQB1 ALLELES IN A SAMPLE OF IRAQI PATIENTS
علاقة مرض حساسية الحنطة بالنمط الوراثي لمستضد كريات الدم البيضاء الصنف الثاني في عينة من المرضى العراقيين

Authors: Hanaa N. Abdullah
Pages: 529-536
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Celiac disease (CD) is a complex disorder triggered by gluten affecting genetically predisposed individuals. The CD is triggered by the binding of one or more gliadin peptides to CD associated HLA class II molecules. Fifty patients with CD and fifty control group were studied. The sera were qualitativelymeasured for anti-TTG-IgA,IgG antibodies and anti-gliadin- IgA,IgG antibodies by ELISA method. The HLA class II (DRB1, DQB1) were genotyped by using Polymerase Chain Reaction-Sequence Specific Primers (PCR-SSP). In the current studypositivity for Anti-TTG antibodies showed a frequency of 38% in CD patients as compared with the control group 0.0%, while high frequency of Anti- gliadin antibodies positivity in celiac disease patient's sera showed 22% as compared with the control group 0.0%with a highlysignificant difference were highly (P=0.001).Human leukocyte antigen genotyping revealed that the DR-alleles, DRB1*03(01,06,08,10), DRB1*0701 and DQB1*02 (01,02) showed highly significant increased frequency in CD as compared withthe controls, while the DRB1*1302 and DQB1*0601 alleles showed significant decreased frequency in CD when compared with the control groups.


Article
GENETIC STUDY ON SWARMING PHENOMENON AND ADHERENCE CAPACITY OF Proteus Mirabilis
دراسة وراثية لظاهرة الانثيال وقابلية الالتصاق لبكتريا Proteus mirabilis

Authors: Wasan W. Al-Bassam
Pages: 537-549
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Five local isolates of Proteus mirabilis(diagnosed in previous study)were selected to carry out studies on genetics depending on their ability to swarming phenomenon and adherence to uroepithelial cells. The results of plasmid content on agarose gel electrophoresis indicate the isolates of P.mirabilis have homogenous plasmid profile and all harbour small plamid bands differ in location and size. Plasmid curing experiments were performed using chemical agents (SDS and ethidium bromide). The results indicated that ethidium bromide caused plasmid elimination of P. mirabilis WEC9 and WEC14 isolates at concentration of 1.25%.WEC14 isolate showed also difference in swarming migration diameter on blood agar after plasmid curing which indicate the involvement of the plasmid and chromosome in regulation of swarming phenomenon in local isolate. The selected isolates indicate the ability for adherence to uroepithelial cells ,and the adhesion average ranged between 32-40 bacteria/uroepithelial cell, in regard of cured isolates the isolates were still able to adhere with an average of adhesion about 29 and 38 bacteria/uroepithelial compared with uncured isolates. The swarming inhibition of Ww14 isolate using various concentrations of anti –swarm agents (Urea, SDS and Tris) was performed. The results showed that Urea at a concentration of 1.25% displayed high inhibition with a mean value of (1±0.6) cm and SDS at a concentration of 1.25% showed inhibition with a mean value of (2 ± 0.7) cm, whereas the results of anti-swarming phenomenon using Tris at various concentrations were not statistically significant.

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