Iraqi Journal of Cancer and Medical Genetics

The present study has been done to assess the mutagenic and antimutagenic effects of Punica granatum (Pomegranate) seed extract against the genotoxic effects ifosfamide (IFO) in male albino mice Mus musculus of BALB/c strain. The pomegranate seed extract (tablets) was provided by Source naturals, INC. Santa Cruse Company, Canada (from Egypt). Two independent experiments were performed. In the first, mice was treated daily and orally with phosphate buffer saline (PBS) as negative control group, and the other three groups were treated daily and orally with three different doses of the P. granatum seed extract (12.5, 25, and 50 mg/kg b.w.) for five weeks in order to test the clastogenetic effects of the plant seed extract. In the second experiment, the first group was positive control group which weekly and intraperitonially was treated with ifosfamide (100 mg/kg b.w.), the other three groups were treated daily and orally with three different doses of the pomegranate seed extract (12.5, 25, and 50 mg/kg b.w.) and also intraperitonially treated at the end of each week with IFO for five weeks to test the protective effects P. granatum seed extract. The results of the first experiment indicated that the P. granatum seed extract has no significant clastogenetic effects on chromosomal aberrations of the bone marrow cells of treated mice. The results of the second experiment was that the P. granatum seed extract, especially at the third dose (50 mg/kg b.w.) exhibited well protective and anti-clastogenetic effect against the genotoxic actions of the ifosfamide on bone marrow cells.

Triple X syndrome 47XXX is a sex chromosome abnormality characterized by presence of an extra X chromosome. Most of the girls born with triple X chromosomes have no signs or symptoms at birth. The condition often remains undiagnosed until adulthood when the genetic defect is discovered on investigation for other reasons as in this case of a 52 year old lady who diagnosed with systemic lupus erythematosus (SLE). The chromosomal analysis for peripheral blood cells by using G-banding technique revealed trisomy X and various abnormality of 3q karyotype. Chromosomal analysis was also done for two daughters and one son of the patient that showed normal karyotype. In conclusion, our findings provide additional support for the hypothesis that X-chromosome polysomy may confer increased susceptibility to SLE. The association of SLE with chromosomal abnormality of 3q found in our patient needs further studies to uncover the role of genes affected by this aberration in SLE.

Cell proliferation characteristics may reflect the aggressiveness of gastric cancer and their eventual prognosis. The aim of this study was to evaluate whether the proliferative activities determined by Ki-67 labeling index (LI) could be used as a predictor in patients with gastric cancer, and examine the relation between COX-2 and Ki-67 expression and gastric cancer. The expression of COX-2 and Ki-67 were detected in 40 patients with gastric cancer and 30 healthy individuals were assessed by immunohistochemistry. The biopsies were fixed in 10% buffered formaldehyde solution, embedded in paraffin and stained immunohistochemically with anti-human COX-2 and Ki-67 antibodies. Expression of Ki-67 and COX-2 was significantly higher in tumor compared with control samples.A significant correlation was found between COX-2 and Ki-67 protein expression. In conclusion, the expression of COX-2 and Ki-67 proteins was closely connected with gastric cancer.

Background: Non Hodgkin lymphoma (NHL) is a diverse group of disorders both in their original history and in response
to therapy .The appearance of Epstein-Barr infection (EBV) in Non-Hodgkin’s lymphoma (NHL) could be distinguished by enzyme link immune sorbent assay by detecting IgM and IgG. The role of EBV as causative agent for non-Hodgkin lymphoma has been supported by identification of large amounts of latent membrane protein 1 (LMP-1) outflow in tumors.
The aim: of this study was to determine the incidence of EBV infection in patient with newly diagnosed patients with non- Hodgkin lymphoma.
Materials and Methods: This study was done at the Department of clinical hematology, the national center of hematology (NCH), Baghdad, Iraq from August 2012 to November 2013. It is a prospective study. A total of 56 patients with age ranged from 18 – 56 years were included. They were diagnosed with NHL and then classified to subtypes by studying histopathology and immunohistochemistry. Assessment of EBV IgM and IgG were done by ELISA technique. A second group comprised of 22 patients with non-malignant condition regarded as control group with matched sex and age. Statistical analysis was performed utilizing SPSS version 17.0.
Result: Mean age of the patients was 51.44±15.93 years. Thirty-six (70.4%) were males and 20 (29.6%) were females. Seven (12.5%) out of 56 cases were positive for EBV–IgM, two cases (28.5%) were follicular lymphoma, 5 cases (71.5%) were diffuse large B cell lymphomas. For the EBV-IgG 12 patients (21.4%) were negative while majority 44 patients (78.6%) were positive, in comparison to control group the result was statistically not significant.
Conclusion: This study showed that the age, sex and clinical characteristics of Iraqi patients with NHL are different from those of surrounding countries and. The incidence of EBV is high in NHL. The most frequent subtype of NHL is diffuse large B cell lymphoma.

Bone marrow derived mesenchymal stem cells (BM-MSCs) represent a promising source for cell therapy, As they can differentiate into bone, cartilage, fat, tendon and many other organ progenitor cells. Although the culture of MSCs has been studied for over 30 years. The identification of standard protocol for isolation and characterization have yet to be developed. A comparison study was made on three different isolation techniques, which include: direct plating, red blood lysis buffer strategies, and density gradient (DG) method (using two different gradient media: Ficoll and Percoll) to find the optimal isolation and culture condition for adequate amount of MSCs for clinical use.The results demonstrate that direct plating of whole bone marrow (BM) suspension provides a suitable alternative protocol for isolation of BM-MSCs with minimal requirements, which mainly based on the frequent medium change in primary culture. The BM crud cell suspension were isolated by aspiration from albino mice and cultured in fresh complete isolation media (CIM) RPMI-1640/10%FBS. Adherent cells from BM cells suspension were MSCs which then expanded by complete expansion media (CEM) α-MEM/ 10% FBS. By concluding, direct plating of whole bone marrow represent the alternative method for isolating of mesenchymal stem cells from small specimen of bone marrow. By concluding, the present study was designed to investigate the optimal technique for the isolation of BM-MSCs for use in tissue engineering and regenerative medicine.

Arginine deiminase (ADI) is an important enzyme in many biological applications and a cancer treatment agent, recently increasing attention is focused to choose a higher productive strain from different bacterial sources. Eighty nine Enterococcus isolates were obtained from 215 samples (20 clinical UTI specimens, 52 human stool specimens, 6 soil and 11 from sewage water), fifty seven isolates were ADI producer. Twenty five isolates were more active in enzyme production, fourteen of them were identified as E. faecium and 11 isolates were E. faecalis. E. faecium isolate M1 obtained from UTI was the most efficient in ADI production, the specific activity of ADI produced from this isolate was 2.64U/mg protein. The effect of culture medium (Mineral salt broth) components on ADI production and other cultural conditions were determined to find the optimum conditions for enzyme production. The maximum ADI production was achieved when the medium was supplemented with 20mM arginine, 1% sucrose, 1% casein, pH 7.5 and incubated at 37ºC for 18 hours. Under these conditions, the specific activity of ADI was 5.1U/mg protein. By concluding, the present study was designed to select ADI higher productive isolate and optimum culture conditions for increasing the production of this enzyme in order to use it as a potent cancer treatment agent after purifying and characterizing it in the future.

Primary amenorrhea which result commonly from sex chromosome aberration as geneticabnormalities, may result from oneaberration in autosomal chromosomes or in association with sex chromosomal aberrations. Chromosomal analysis was performed for the 26 years old lady (married and infertile) with primary amenorrhea by using G-band technique. Four autosomal chromosomes are involved in producing primary amenorrhea with normal phenotype, secondary characteristics were associated with mosaic karyotypes the
1st: 46,XX,(3;5;10)(q13-23;q23-35;22-25),(5;19)(qter;p13.2),del(19p13.2).
2nd: 45,XX, ,(3;5;10)(q13-23;q23-35;22-25),del(12),del(12),del(19p13.2),(M).
The sex chromosomes look normal.The conclusion was that may be some autosomal chromosomes regulate reproductive development and may the alerted regions on chromosomes in recent study have effect somehow on regulation of reproductive development.

Bone marrow derived mesenchymal stem cells (BM-MSCs) were successfully used in regenerative medicine. The purpose of this study was to isolate and characterize mesenchymal stem cells from mouse bone marrow for their subsequent use in researches. Previous data suggest that BM-MSCs are typically enriched by plastic adherent cultures, fibroplastoid cell fraction. However, Identification of MSCs achieved through their morphology, phenotypic characteristics and their biological behavior. The cellular morphology played a major part in identifying MSCs in vitro. In general, immature MSCs appeared as small, spindle-shaped cells, whereas mature MSCs was displayed as larger cells with a flat, polygonal morphology. Cells also tended to be locally confluent, growing in distinct colonies. Immunophenotypic analysis demonstrated that mouse BM-MSCs at passage three uniformly positive for CD44, CD90, CD105 and CD106. However, MSCs were always found to be negative for heamatopoietic specific markers CD34, CD45, and endothelial marker CD31. By concluding, mesenchymal stem cells can be successfully derived from mouse bone marrow by direct plating method. These cells represent a valuable source of stem cells for restoring the damaged organs.

This study aimed to investigate the ability of Iraqi strain of Newcastle Disease Virus (NDV) to induce apoptosis in vivo through Caspase 3 activation when administered intratumoral and compared to intra-peritoneal injection. Immunohistochemistry test was used for detecting apoptosis by using mAb against Caspase-3. Histopathological sections for the treated tumor mass showed proliferation of granulation tissue with extensive area of necrosis (mixture of apoptotic and necrotic cells). This revealed that NDV infection induce apoptosis significantly in mammary adenocarcinoma (AM3) when compared to control group proved by high expression of caspase-3.In vivo immunohistochemical detection of Caspase 3in mammary adenocarcinoma which give brown stain revealed a significantly(p<0.05)increase in the mean percentage of cells expressing caspase 3 in NDV treated group compared with low increase in the mean percentage of cells expressing caspase 3 in untreated control group at day 1,2,3,7, and at 14 day. These results revealed that NDV had powerful effect on inducing apoptosis in mammary adenocarcinoma (AM3) during its replication inside the tumor mass for long time after one single injection. This study indicate the role of NDV Iraqi strain in inducing apoptosis as confirmed by caspase-3 activation in cancer cells which is interesting confirmed feature that make NDV Iraqi strain as anti-tumor agent.

Acute lymphoblastic leukemia (ALL) accounts for about 25 percent of all childhood cancers and 75–85 % of all childhood leukemias. Thirty six children with acute lymphoblastic leukaemia were tested for antibodies to Epstein-Barr (EBV) virus with age ranged (2-12) years (20 males and 16 females) attending pediatric central hospital, oncology-hematology department. Thirty children with age ranged (2-12) years were regarded as control group. The two groups undergo Epstein-Barr virus antibodies assays (EBV-EAIgG, IgM, EBV-VCAIgG, and IgM) by enzyme linked immunosorbent assay (ELISA). The samples analysis was observed that the percentage of VCA-IgG (44.4%), VCA IgM (2.7%), EA IgG (27.7%) and EA IgM (2.7%) were higher than control group. The mean values of EBV (VCA IgG) in ALL group increase significantly when compared with control group, while no significant difference between ALL and control in the mean values of VCA IgM. There was a significant increase in the mean of EA IgG in ALL than in control group, whereas, no significant differences were noticed in EA IgM between two groups. These results clearly indicated that provide a working hypothesis and some facts on which further investigations of etiology of ALL be planned.

This study describes utilization of large silver nanoparticles (above one hundred nanometer in diameter) synthesized by plus laser ablation method in liquid (PLAL) in growth inhibition and apoptosis induction of cancer cells. Two different types of cancer cell lines and two different types of transformed cell lines were used. These cell lines were Rabdomyosarcoma (RD), glioblastoma (AMGM), rat embryo fibroblast transformed cell line (REF) and green monkey kidney transformed cell line (VERO). Cells were challenged with diluted concentration of silver nanoparticles they were 62.5μg/ml, 30.25μg/ml, and 15.12μg/ml. The synthesized silver nanoparticles was characterized it had surface plasmon resonance peak at 411nm and particle size distribution average diameter of 193.19 nm. Growth inhibition assay revealed that these relatively large nanoparticles are lethal toward cells under investigation; this lethality was concentration dependant during 24 hours of incubation at 37°C. According to acridine orange/propidium iodide mixed fluorescent staining assay, the used concentrations were able to induce apoptosis in all cells tested. However, the highest concentration was able to induce highest apoptosis percentage in the treated cells. This was confirmed by DNA fragmentation assay as well. In conclusion this study determined that above-one hundred nanometer size silver nanoparticle is capable of inducing apoptosis in cancer cell lines.

Klinefelter’s syndrome charactrised by the presence of one or more extra X chromosome in males and it is represent the most common chromosome aneuploid abnormality usually detected in men. Several types of research works have done on this syndrome in different countries worldwide to reveal many features of this syndrome including genetics, behavior, fertility, age, health problems and others. Almost all Klinefelter’s syndrome men are infertile, tall, depressed, autistic, violent, tending to make crime, disappointed, learning difficulty etc. In Iraq Klinefelter syndrome has not been studied scientifically therefore this study has been designed to put first steps to reveal some features of this syndrome in Iraq. Chromosomal analysis and questionnaire were performed on men who suffered from infertility and on prisoners. Chromosome studied was done by using G-banding technique on all cases to verify Klinefelter’s syndrome. The study included 823 male who showed infertility and 217 prisoners. The results showed that 15% (126 cases) of infertile men have Klinefelter’s syndrome, 77% (95 cases) out of them were (47, XXY), while 23% (29 cases) were mosaic for Klinefelter’s syndrome (46,XY/47,XXY). Moreover, 97% (122 cases) of Klinefelter’s bearing men were tall. Seven men (3%) of the prisoners showed Klinefelter syndrome, no mosaicism was detected. In the prisoners group, 27 persons were tall and all the seven prisoners with Klinefelter’s syndrome were among them to form 26% of them. Three of those seven prisoners were married but having no children. Three out of those seven were 20 years old and below. The prisoners with Klinefelter’s syndrome did not show remorse and most of them justified their crime as rehabilitation response. Although the number of cases involved in this study was not large, the results clearly revealed that Klinefelter’s syndrome is markedly occurred in Iraq particularly in infertile men and in prisoners. However this syndrome has not drawn any attention and has not been considered correctly through medical assessments. This study strongly recommends doing multidisciplinary research works on this syndrome through large epidemiological study through focusing on health, social, behavior, psychological and genetic of men who have this syndrome. Moreover this study recommends performing chromosomal analysis for every newborn, infertile, and tall prisoner males.

In this study, investigation done on the effect of crude aqueous extract of Achillea Fragrantisime on the cytotoxicity of different cancer cell lines . This study containing three parts, first one including selection of four types of different cancer cell lines, two types of these are cancer cell line which are (AGM-3) and (AMN-3) and the other tow types are transformed which ( REF) and (VERO) for measuring the cytotoxicity of crude extract on them, The results showed that the effect of crude aqueous extract of this plant depends on the type of cell line concentration of extract and exposure time , it was found in cancer cell lines that the high concentrations of extract investigating the highest inhibition percentage of growth cell when compared with Control, while this percentage was less in inhibition of growth cell at low concentrations after 24 hours and then this percentage be lower after exposure for 48 hours, while after exposure for 72 hours there was no significant differences when compared with control. Same results where appeared in transformed cell line but the inhibition percentage was less from what is found in cancer cell line.
Second part of this study has been measured the effect of plant crude extract on form and stimulate an apoptosis in cancer cell lines and that was happened through application of Tunnel assay technology and, the results appeared that this crude extract was stimulated on Apoptosis process in all used types of cancer cell lines . Finally, at the third part of this study was included measurement of genetic toxic effect of the plant crude extract through analysis micronucleus assey in human lymphocyte stimulated by Phytoheamaglotenine, and the Results appeared lack of MN which is one of certified analysis to investigation of breaking genetic material.
The conclusion of the study that the crude aqueous extract of Achillea Fragrantisime was useful in inhibition of growth cancer cells, and increases stimulate appaptosis process , as well as it non toxic in normal lymphocytes and that make it outbalance to use it as alternative treatment to chemotherapy and radiotherapy habitual use in spite of there side effects and some disadvantage.

Caramel food colorings which are used widely in products and processed foods, as used in the manufacture of sweets and soft drinks, and in recent years has raised a lot of doubts and warnings about its use. The fact that these pigments have direct contact to human health, especially children, and due to the lack of studies illustrate precisely the toxic effects and genetic these colorings, this study was to shed light on the effect of cytogenetic for dyes caramel ( Category IV ), in different concentrations ( 5,10,15,20 ) μg / mL, blood cells in human lymphocytes (in vitro) Using cytogenetics Parameters (Mitotic Index(MI), chromosomal aberrations(CA), and Micronucleus (MN). These results indicate that the colorings caused chromosomal changes consisted of The Chromosome Aberration represented by chromosomal and chromatid breaks, chromosomal and chromatid gap, and dicentric chromosome, also caused a reduction in( Mitotic Index(MI) and increase of micronuclei.