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: 2010 volume:9 issue:2

Article
PURIFICATION OF MURINE HEAT SHOCK PROTEIN 90-BETA (84 KDA) FROM MICE TUMOR CELL LINE
تنقية بروتين الصدمة الحرارية 90 نمط بيتا(84 كيلودالتون)الفأري من أحد خطوط خلايا سرطان الفئران

Authors: عامر طالب توفيق
Pages: 124-136
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Abstract

Murine heat shock protein 90 beta isotype (84 kDa) was purified from the cytoplasm of AMN3 cells (locally established mice mammary adenocarcenoma cell line). This protein does not have an in vitro activity assay, to confirm its presence during purification, detection of protein with molecular weight equal or near 84 kDa was followed in each purification step. The cell cytoplasm fraction of AMN3 cells contained proteins of molecular weight (MW) ranged from 23.4 to 85.1 kDa as determined with SDS-PAGE analysis. The precipitate of 70% ammonium sulfate saturation of cytoplasm fraction exhibited protein bands with molecular weight ranged from 31.6 to 93.3 kDa. Ion exchange chromatography with DEAE-Cellulose column for the ammonium sulfate precipitated cytoplasm proteins produced one prominent peak, eluted between 0.5 and 0.7 M NaCl. This peak contained four proteins with MW ranged from 17.3 to 85.1 kDa. Gel filtration through Sepharose 6B column for the peak eluted from DEAE-Cellulose column was conducted, different peaks were resulted. A peak with estimated molecular weight of 87 kDa was selected. Second round of gel filtration through Sephacryl S-200 was conducted for the 87 kDa selected fractions eluted from the previous gel filtration step. The presence of one peak was emphasized and its estimated MW was 83.176 kDa. The molecular weight and purity of this protein were checked again with SDS-PAGE. The identity of the purified protein as Hsp90β was confirmed with immunofixation assay by gel electrophoresis on 1% agarose using standard monoclonal antibody specific for murine Hsp90β.

Keywords

HSPs --- Hsp90 --- cancer cell --- immunotherapy


Article
CLINICAL OBSERVATION OF CYTOKINES IN PATIENTS WITH HEPATITIS B
ملاحظة سريريه للسايتوكينات في مرضى التهاب الكبد الفيروسي البائي

Authors: آمنة نعمة الثويني1
Pages: 137-143
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Abstract

Thirty serum samples were collected from patients with chronic hepatitis B who were admitted to Hepatology and Gastroenterology Teaching Hospital in Baghdad during period from beginning of February / 2008 to the end of February / 2009, also serum samples were collected from 30 healthy HBs Ag carrier was discovered accidentally through attending blood bank for donation of blood and a total of 30 apparently healthy blood donors were included in the study served as a control group. The concentrations of some cytokines Pg/ml (IFN-γ, GM-CSF, IL-1α, IL-6, IL-8 and IL- 2 receptor) have been estimated in the sera of the studied groups and the results showed that mean levels of gamma-interferon, interleukin-1 alpha, interleukin-8 and interleukins-6 in sera of carriers group were (109.6, 251.6, 232.9 and 166.8 respectively) more than the level of these cytokines in sera of chronic HBV patients (75.1, 94.7, 159.6 and 110.9 respectively) and healthy control (48.4, 10.1, 11.3 and 15.4 respectively), while the mean level of GM-CSF was elevated in sera of CHB patients (110.8) than both of healthy control and carrier group (17.3 and 41.2 respectively).Finally the interleukin-2 receptor was represented with similar level in sera of chronic patients and carrier group ( 165.6 and 167.9 respectively) with non significant difference, whereas all other statistical comparisons among the three main groups showed highly significant difference.

Keywords

Hepatitis --- CHB patient --- Cytokines.


Article
ESTIMATION THE SERUM CONCENTRATION OF INTERLUKINE 2 RECEPTORS IN PATIENTS WITH RHEUMATOID ARTHRITIS
قياس تركيز مستقبلات انترلوكين -2 في مصول مرضى التهاب المفاصل الريثاني

Authors: آمنه نعمه الثويني
Pages: 144-149
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Abstract

Serum soluble interleukin 2 receptors (sIL-2R) concentration reflect lymphocyte activation in vivo. The present study was conducted to determine if sIL-2R concentration correlate to disease activity parameters in patients with rheumatoid arthritis (RA) compared with healthy populations. One hundred blood samples were collected, from RA patients and thirty from apparently healthy groups. The mean age of RA patients was 46.75±12.82 and, the majority of patients were females (84%). The level of sIL-2R was estimated serologically by using enzyme linked immunosorbant assay. The result indicated that the sIL-2R was elevated significantly (mean 2101.67± 1899.21) in RA compared with healthy control group (mean16.76±5.75).


Article
EXTRACTION AND PURIFICATION OF MANNOSE-SPECIFIC LECTIN FROM
استخلاص وتنقية اللكتين المتخصص للارتباط بالمانوز من بكتريا Escherichia coli

Authors: امنة نعمة الثويني
Pages: 150-157
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Abstract

This study included isolation and Purification of Mannose-specific lectin (type 1 pili) from E. coli strain isolated from a human urinary tract infection. The pili were removed from the bacteria by mechanical shearing, precipitated out of solution by ammonium sulfate at a final concentration of (20%). The purification of the pili was carried out by ion exchange chromatography. There was one identical peak correspondence with the hemagglutination activity of the lectin with guinea pig erythrocytes. The further purification procedure was achieved by using gel filtration on Sepharose 6B column.


Article
THE EFFECT OF ASSISTED REPRODUCTIVE TECHNOLOGY AND SPERM ACTIVATION TECHNIQUES ON SEMEN CHARACTERISTICS AND PREGNANCY RATES
تأثير التقنيات المختبرية و تنشيط النطف المساعدة على الإنجاب في متغيرات تحليل السائل المنوي ومعدلات الحمل

Authors: باسم خميس الركابي
Pages: 158-166
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Abstract

This study was designed to assess the effect of sperm activation techniques In vitro on semen characteristics and intrauterine insemination (IUI) procedure in varicocelic infertile patients. In this study, fifty infertile male involved and according to preparation methods divided into 25 varicocelic and 25 non-varicocelic infertile couples. From each male, semen samples were analyzed before and after in vitro sperm processing by standard semen parameters. However, the spermatozoa prepared by direct swim-up and centrifugation swim-up technique with addition of IVF culture medium. Furthermore, sperm concentration, sperm motility, progressive sperm motility and normal sperm morphology were evaluated according to WHO standard criteria. For IUI technique, sperm prepared and incubated for 30 minute in 5% CO2 at 37ºC. The results of the present study indicate a highly significant (P<0.001) differences for all sperm functions and pregnancy rates (P<0.001) were reported post in vitro sperm activation using direct swim-up and centrifugation swim-up method as compared to pre-activation. Whereas, the best results for clinical pregnancy rate were reported for semen samples prepare by using direct swim-up when compared with centrifugation swim-up method. It was concluded that centrifugation force have detrimental effect on sperm plasma membrane activity and fertilization potential of human spermatozoa. Further studies are recommended to evaluate the detrimental effect of centrifugation swim-up technique on DNA damage and embryo quality after in vitro fertilization and embryo transfer.


Article
STUDY THE INH INHIBITORY EFFECT OF Lactobacillus acidophilus ISOLATED FROM YOGHURT AS PROBIOTICS ON Candida albicans GROWTH IN VITRO AND IN VIVO
دراسة التأثير التثبيطي للعصيات اللبنية المعزولة من اللبن الرائب كمعززات حيوية للحد من نمو خميرة Candida albicans خارج و داخل الجسم الحي

Authors: بشرى جاسم محمد
Pages: 167-179
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Abstract

This research was designed to study the inhibitory effect of Lactobacillus acidophilus which has been isolated from yoghurt as probiotics in reduction of Candida albicans growth in vitro and in vivo. The results showed that the Minimum Inhibitory Concentrations and Minimum Fungicidal Concentrations were 60% and 70% respectively, also the inhibition zone of L. acidophilus against C. albicans reached to 26mm in solid medium .The out come of in vivo study (histological examination ) clarified that C. albicans caused clinical pathological effect in mice tissue organs ( liver , intestine, stomach , kidney) when administrated orally by 1.5x 10 8 cfu ml C. albicans that effect decrease by orally inoculated with same dose of L. acidophilus.The results reflect the ability of L. acidophilus to reduce certain clinical pathological change in mice organs, with promising encourage to use the L. acidophilus as biotherapeutic agents against C. albicanas infections.


Article
ASSOCIATION BETWEEN HLA-CLASS II ALLELES AND T-CELL PROLIFERATION IN RESPONSE TO ENTEROVIRUSES AND ADENOVIRUS ANTIGENS IN NEWLY DIAGNOSED CHILDREN WITH TYPE 1 DIABETES MELLITUS
العلاقة بين مستضدات التطابق النسيجي- الصنف الثاني والفعالية الوظيفية للخلايا اللمفية بعد تحفيزها بمستضدات الحمات المعوية والحمات الغدية في الاطفال المصابين حديثاً بالسكري من النوع الأول

Authors: ايمان مهدي صالح
Pages: 180-190
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Abstract

Viruses may be involved in the pathogenesis of Type 1 Diabetes Mellitus (T1DM), either through direct β-cell infection or as triggers of autoimmunity. T- cell proliferation was evaluated in response to Enteroviruses antigens including Coxsackievirus B and Poliovirus in addition to Adenovirus in an HLA- matched population of children with T1DM and in children who were apparently healthy. A total of 60 Iraqi T1DM children were included in the presents study. They were with new onset of the disease. For the purpose of comparisons, 50 apparently healthy control subjects were also selected. HLA typing was measured by microlymphocytotoxicity, while methylthiazoltetrazolium (MTT) assay was used for lymphocyte proliferation by culturing peripheral blood lymphocytes (PBL) with Coxsackievirus B5, Adenovirus 3, 4 and 7 and Poliovaccine. No significant differences were shown in the PBL proliferative percentage in response to Con-A mitogen and tested viruses (CVB5 and Adenovirus) between T1DM and healthy controls, but PBL proliferative percentage of patients showed a significant decline in response to Poliovaccine. HLA class II (-DR3, DR4, DQ2 and DQ3) antigens were significantly increased in T1DM patients and they played an important role in the etiology of the disease. Strong T-cell proliferation in response to the tested viral antigens were observed to be related to HLA-DR4 and HLA-DQ3 antigens, whereas the HLA-DR3 and HLA-DQ2 alleles were associated with week responsiveness to the same antigens. However, in children with new- onset diabetes, responses were decreased and this could be caused by trapping of virus- specific T- cells in the pancreas.


Article
BIOCHEMICAL STUDY ON SUPEROXIDE DISMUTASE ENZYME IN PATIENTS WITH DIFFERENT BRAIN TUMORS
العلاقة بين مستضدات التطابق النسيجي- الصنف الثاني والفعالية الوظيفية للخلايا اللمفية بعد تحفيزها بمستضدات الحمات المعوية والحمات الغدية في الاطفال المصابين حديثاً بالسكري من النوع الأول

Authors: حذامة رزوقي حسن
Pages: 191-201
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Abstract

The aim of the current study is to in evaluate the role of SOD activity in the previously reported oxidative stress in our laboratory(1), in the patients with different brain tumors. SOD activity was assayed according to riboflavin/NBT method and its specific activity was calculated in patients with benign and malignant brain tumors and control. Moreover the specific activity was compared in these samples according to gender and the occurrence of disease.Non significant elevation (P > 0.05) in SOD specific activity was observed in tissue of malignant tumors in comparison to that of in benign brain tumors. While a highly significant decrease (P < 0.001) of the specific activity was found in sera of malignant patients group in comparison to that of the control group, and it was found lower in female than male in control and malignant groups. An elevation in this specific activity was noticed in patients with secondary brain tumors in comparison to that of primary brain tumors (P<0.05). From the results of the present study we conclude that the observed decrease in SOD activity in sera of patients with different type of brain tumors contribute to the oxidative stress that previously reported in our laboratory to be present in such patients.


Article
EFFECT OF 1% HYDROGEN PEROXIDE (H2O2) IN DRINKING WATER ON SOME PARAMETERS IN ِِADULT MALE RABBITS
تأثير1% بيروكسيد الهيدروجين في ماء الشرب على بعض المعايير الكيموحيويه في ذكور الارانب البالغة

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Abstract

The present study was undertaken to investigate the effect of 1% of H2O2 on some biochemical markers related to oxidative stress,cardiac, hepatic and thyroid functions in adult male rabbit's.Twenty adult male rabbits were divided randomly into two groups (10/group), the first (control) group and the second (T) group: animals were received 1% hydrogen peroxide (H2O2)in drinking water for one month.At the end of the experiment, fasting blood samples were drawn by cardiac puncture technique from all experimental animals for measuring : a- serum concentration of total cholesterol(TC);triacylglycerol( TAG);low density lipoprotein –cholesterol( LDL-C) ;very low density lipoprotein –cholesterol (VLDL-C) and high density lipoprotein –cholesterol (HDL-C) b- serum alanine transaminase (ALT) ;aspartate transaminase (AST) and lactate dehydrogenase (LD) activities c- albumin and globulin concentrations. d-Platelets count and prothrombin time e-serum glutathione (GSH) and malondialdehyde (MDA) concentrations. F-serum thyroid stimulating hormone (TSH), triiodothronine (T3),tetraiodothyronine(T4) and glucose concentrations. The results revealed that male rabbits receiving 1% H2O2 in drinking water for one month showed a significant increase(p<0.01) in serum TC, TAG, LDL-C, VLDL-C and MDA concentrations, ALT, AST and LD activities and platelets count, and a significant decrease (p<0.05) in serum HDL-C, GSH, albumin, globulin concentrations and prothrombin time comparing to control, besides significant decrease (p<0.05) in serum T3 & T4 and significant increase (p<0.05) in serum TSH, and glucose concentration were observed. On conclusion, it has been found that exposure to1% H2O2 of exerts deleterious effect on cardiac and hepatic functions, hypothyroidism and state of oxidative stress.


Article
PURIFICATION AND CHARACTERIZATION OF CHOLERA TOXIN FROM CLINICAL ISOLATE OF Vibrio cholerae
تنقية وتوصيف ذيفان الكوليرا من عزلات مرضية لبكتريا Vibrio cholerae

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In this study fifteen diarrhea isolates suspected as V. cholerae were obtained from Central Health Laboratory/ Ministry of Health and their identification indicated that four isolates were belonged to O1 serotype and designated as S,18,22,13 and all from El Tor biotype, while the fifth isolate was serogrouped as non-O1 serotype and designated as V. cholerae ab. Heat labile enterotoxin (LT) were determined in all clinical O1 and non-O1and the purification scheme was improved; few steps were adopted to include back extraction of Ammonium Sulfate saturation between 20-80%, desalting through Sephadex G-25 and gel filtration by Sephadex G-100. The specific activity was increased to 100 fold at the final step. The toxin purity indicated no other protein detected in toxin preparation by employing gel filtration and SDS-PAGE. The toxin had a molecular weight of 83.17 KDa as indicated by gel filtration. The stability of purified CT at different temperature proved that CT could be stored at -4°C for 6 month without loss in bioactivity, while remaining activity (%) reached to 35% and 20% after 30min incubation at 45°C and 55°C and 65°C respectively. The bioactivity of toxin diminished at 100°C for 10 min. The remaining activity (%) of purified CT treated with different PHs showed CT susceptibility to low pH (2 – 4) moderate stability at pH 7 and high stability at pH 8 - 9.


Article
RADIOLOGICAL AND CHEMICAL HAZARDS OF PUBLIC EXPOSURE TO URANIUM-235 AT AL-TWAITHA NUCLEAR RESEARCH SITE
تقدير المخاطر الإشعاعية والكيمياوية الناجمة عن التعرض لليورانيوم-235 في موقع التويثة للأبحاث النووية

Authors: نبيل هاشم التميمي
Pages: 226-238
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Abstract

Multi-step risk assessment process is used in this study to characterize the possible health hazards caused by exposure of the local inhabitants living in the vicinity of Al-Twaitha nuclear research site to uranium-235 through environmental pathways (inhalation of air and ingestion of contaminated foodstuffs) using a hypothetical linear no-threshold (LNT) statistical model. Terrestrial food chain mathematical models are used in this study to predict the uranium-235 transfer from soil to agricultural crops (vegetables) and to animal products (meat, milk, milk products and egg) consumed by humans. The possibility of developing fatal cancer to Diyala bridge and Eshtar population as a consequence of uranium-235 exposure is evaluated at 0.14 latent cancer fatalities per million exposed individuals. The risk of developing breast cancer is found to be occurs at a largest extent (0.0135%) in comparison with cancers to other body organs and tissues. Other consequences of radiation injury (genetic damage transmitted to succeeding generations) are expected to occur at a rate of 1850 per million exposed individuals in the offspring of Jesr-Diyala population as a result of changes transmitted via the genetic mechanisms due to irradiation of gonads, or there is 1 extra abnormally case in a group of 540 parents who were irradiated before conception occurred.


Article
PURIFICATION OF KLEBOCIN EXTRACTED FROM LOCAL ISOLATE Klebsiella pneumonia (K64)
فصل وتنقية الكليبوسين من العزلة المحلية Klebsiella pneumonia (K64) واختبار فعاليته ضد المايكروبية

Authors: رشيد محجوب مصلح
Pages: 239-247
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Abstract

Klebsiella pneumonia (K64) isolated from urine was induced by mutagenic agent (Mitomycim C) to produce Klebocin. The Klebocin was extracted and ultra filtrated then purified by Ion-exchange chromatography. Klebocin was obtained by Gel-filtration chromatography; the purified protein was applied to SDS-PAGE gel electrophoresis. The antimicrobial activity was tested against different microorganisms that included: Escherichia coli, Klebsiella pneumonia, Staphylococcus aureus, Candida albicans and Aspergillus niger. The Klebocin partially purified from its immunity protein by ion exchange chromatography and completely purified by gel filtration chromatography. The purity was confirmed from one band obtained from electrophoresis. Staphylococcus aureus was very sensitive while (K72) K.pneumonia revealed a minimum inhibition zone

Keywords

Klebocin --- Klebcin (K64)


Article
STUDY THE PATHOLOGICAL EFFECTS OF COVERED SMUT INFECTED WHEAT IN ALBINO MICE
دراسة التأثيرات المرضية للحنطة المصابة بالتفحم المغطى في الفأر الأبيض

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One hundred kg infected wheat, Triticum astivium were collected from infected fields in Jalaola in Iraq during winter (2007-2008). The infected plants were characterized according to its fishy odor and the ratio of bunt balls. The fishy odor referred to the infection by common bunt spores which produce trimethylamine. The ratio of bunt balls was found to be more than 30 bunt balls in 250g, therefore it designated as "smutty". The spores which were taken from the infected grains found that they were Tilletia careis and Tilletia foetida species. The presence of trimethylamine in aqueous extract of infected grains was determined by using different chromatography techniques, which confirmed the presence of trimethylamine. Forty albino mice were used for studying the pathological changes. These animals were divided equally into 2 groups 20 mice were fed for 90 days infected wheat pellets administrated orally. The pathological changes were recorded at periods 20, 40, 60, and 90 days respectively. The 2nd group considered as control group which fed pellets without infected wheat. The pathological changes appeared during 20, 40, 60 and 90 days in liver, kidney and spleen but pathological changes in heart occurred after 60 days of administration and after 40 days in lungs whereas in intestines occurred after 20days of administration. Liver and spleen were showed enlargement with acute cellular degeneration and multifocal granulomatous changes. In conclusion, common bunt infected wheat is harmful and causing tissues damage.


Article
BINDING OPTIMIZATION OF AFP WITH ITS ANTIBODY BY IRMA METHOD IN TISSUE HOMOGENATE OF SOME OVARIAN TUMORS
الظروف المثلى لارتباط البروتين الجنيني ألفا مع مستضده باستعمال طريقة الاختبار المناعي الإشعاعي المتري في مجانسات بعض أورام المبايض

Authors: سامي المظفر
Pages: 258-273
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Immunoradiometric assay (IRMA) was modified and used to characterize the binding of 125I-anti AFP antibody with its AFP antigen of benign germ cell (OI), malignant germ cell (OII), malignant serous (OIII) and isolated AFP from malignant germ cell ovarian tumor tissues. Different factor′s (concentrations of antibody, concentration of antigen, pH, time, temperature, and different concentrations of ions) affecting this binding were extensively studied. The observed optimum conditions for the binding were as follow: For (benign germ cell OI, malignant germ cell OII, malignant serous OIII and for isolated AFP of malignant germ cell), the optimum protein concentration in tissue homogenate was (300, 200, 200 and 75mg.ml-1), respectively. 125I-anti AFP antibody optimum concentration was (500, 400, 500 and 300mg.ml-1), respectively. Optimum pH was (8.0, 8.0, 7.6 and 8.0) respectively. The optimum time and temperature were 120 minute at 25˚C for (OI, OII, OIII) and180 minute at 4˚C for isolated AFP of malignant germ cell. The use of different divalent cations showed a decrease in the binding of AFP with its antibodies except for Mn+2 in OI, OII and OIII. In addition to a decreases the binding of AFP with its antibodies by using different halides in OI, OII and OIII.


Article
DETECTION OF EPISTEIN BARR VIRUS IN BREAST CANCER USING THE POLYMERASE CHAIN REACTION AND IN SITU HYBRIDIZATION
الكشف عن فيروس أبستين – بار في سرطان الثدي بإستعمال تقنية سلسلة تفاعل البلمرة و التهجين الموضعي

Authors: سجى جهاد الخالدي
Pages: 274-285
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In order to prove the implication of Epstein-Barr virus in breast cancer of Iraqi patients, breast cancer tissues were collected from 50 newly diagnosed patients recruited at The Medical City Teaching Hospital and Al-Helal Al Ahmar Hospital in Baghdad. Twenty non-cancer breast tissue biopsies represented the control group. The polymerase chain reaction was applied to detect Epstein-Barr nuclear antigen-1 (EBNA-1) and latent membrane antigen (LMP-1) viral genes. Latent Epstein-Barr virus encoded RNAs (EBER 1 & EBER 2) were detected using in-situ hybridization technique. The relation to breast cancer type and grade were studied. The amplified EBNA-1 bands of EBV were identified in 26/5 (52%) and the amplified LMP-1 bands were identified in 10 /50 (20%) of the malignant group. Positive in situ hybridization signals for EBER-1&2 were detected in 6/50 (12%). The presence of the virus was proved by EBNA-1 and LMP-1 in 6 (12%) cases, by EBNA-1 and EBERs in 4 (8%) cases, by LMP-1 and EBERs in 2 (4%) cases and by all markers in 4 (8%) cases. It was concluded that the relation of this virus to breast cancer is potentially important as an etiological factor. It also helps identifying women at risk for this type of cancer. It could assist early diagnosis and measurement of post – therapy residual disease.


Article
ROLE OF VOLATILE OILS FRACTION OF Cyperus rotundus L. IN INDUCTION OF APOPTOSIS ON CANCER CELL LINES IN VITRO
دراسة دور الزيوت العطرية لعشب السعد في استحثاث عملية الموت المبرمج في مختلف الخطوط الخلوية السرطانية

Authors: زيد عبد المنعم علي1
Pages: 286-298
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The induction of apoptosis was evaluated in the current study, and it was found that volatile oils extracted from Cyperus rotundus L. suppressed growth and proliferation of the three cancer cell lines (Human Epidermoid Larynx Carcinoma (HEp-2), Murine Mammary Adinocarcinoma (AMN-3), Human Glioblastoma Multiformi (GBM)) via early induction of apoptosis (loss of mitochondrial membrane potential). Characteristic DNA fragmentation (late apoptosis) of treated cancer cells with the oils revealed that a definite induction of apoptosis and this may due to the cytotoxic effect of that extract. In both early and late induction of apoptosis tests, REF cells did not expressed any changes in its cellular behavior after treatment with the C. rotundus extract. Induction of apoptosis was further confirmed by investigating the changes in a mammalian cell cycle. It was observed that the volatile oils of C. rotundus L. extracts caused a significant cell cycle arrest at G2/M transition phase, due to the ability of that extract to suppress the phospho-cyclin dependent cyclase-2 (cdc-2) which is responsible for moving the cell cycle from G2 to M phase. Therefore the cells will be obligated to be arrested and undergo apoptosis, but the same extract did not expressed any change in its cdc-2 level after REF cells treatment, therefore the cells were not arrested at G2/M. Finally it was concluded that the volatile oils C. rotundus enhanced the induction of apoptosis on cancer cell lines. ___________________________________________________________________

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