Radiation-induced genomic instability and cellular communication: mechanistic investigations

Communication between irradiated and un-irradiated (bystander) cells can cause damage in cells that are not directly targeted by ionizing radiation (IR); a process known as the bystander effect (BE). BE can also lead to genomic instability (GI) within the progeny of bystander cells, similar to the progeny of directly irradiated cells. The molecular factors that mediate this cellular communication can be transferred between cells via gap junctions or be released into the extracellular media/microenvironment of cells and tissue following irradiation. Although GI is thought to be a critical step in the onset and progression of cancer, BE response contributions in such processes are still not clear. Therefore, this study was designed to investigate the risks or benefits associated with the induction of non-targeted effects especially BE following exposure to low LET X-ray radiation using two different cell types. Additionally, the project aims to achieve an increased understanding of the mechanisms of non-targeted effects of ionizing radiation by examining the molecular signalling via exosomes within the irradiated, bystander and progeny of irradiated and bystander cell population. Different cell combinations were established between tumour (MCF7) and nontumour (HMT-3522S1) human breast epithelial cells using a 6-well plate co-culture system. The cells were irradiated with two doses of X-ray; 0.1 Gy (a iagnostic procedure relevant dose) and 2 Gy (therapeutic dose) and a sham-irradiation dose of 0 Gy (for control groups of experiment). The co-culturing time was 4 hours for all cell combination, whereupon a media transfer approach was used to induce BE within the cells in the exosome part of this study. The early and late cellular damage responses were evaluated by the following biological endpoints: cytogenetic/chromosomal analysis, apoptotic analysis, telomere length and telomerase activity measurements. In addition to these biological endpoints, the comet assay was utilised to estimate the initial and delayed DNA damage within the cells that had been treated with exosomes, previously extracted from the irradiated, bystander and control cell media. The results showed that 2 Gy direct irradated MCF7 and HMT cells were both able to induce early and late chromosomal damage in the bystander MCF7 and HMT cells. Furthermore, these bystander cells exhibited early and delayed telomeric instability, which could prompt further GI at later time-points. In comparison, 0.1 Gy direct ii irradiated MCF7 cells were only able to induce initial and delayed chromosomal damage within the bystander MCF7, which also demonstrated a high level of telomeric instability at early and late time-points. While, bystander HMT cells did not show chromosomal damage after 1, 12 and 24 generations/population doublings following co-culture with 0.1 Gy direct irradiated MCF7 or HMT cells. 0.1 Gy bystander HMT cells did reveal a high level of apoptosis at early and late time points, which might be due the removal of cells with a high level of chromosomal damage. Interestingly, the 0.1 Gy bystander HMT cells exhibited significant levels of telomeric instability at early and late time points, which could contribute to chromosomal instability at later time-points. The investigation in to the mechanisms of molecular signalling via exosomes showed that the exosomes of irradiated cell conditioned media (ICCM) from MCF7 cells had the ability to induce BE within MCF7 and HMT cells similar to the effects of ICCM following 2 Gy X-ray. The exosomes that were isolated from the MCF7 bystander cell media had a similar effect as the ICCM on the MCF7 and HMT bystander cells. These exosome-bystander cells also showed GI within their progeny after 24 generations and retained the ability to induce cellular damage to fresh un-irradiated MCF7 cells, demonstrating an underlying mechanism for propagating the delayed damage responses. The inhibition of the exosome’s cargo molecules by RNase treatment and protein denaturating (boiling of exosmes) significantly abrogated BE and GI in both MCF7 and HMT bystander cells following 2 Gy X-ray. Thus data demonstrated crucial roles for exosome RNA and protein molecules in the non-targeted effects of IR induction. In summary, our investigations demonstrate that BE has detrimental consequences within the tumour and non-tumour breast epithelial cells (MCF7 and HMT3522S1) following low and high doses of X-ray irradiation, and these detrimental consequences are frequently mediated by exosomes that contain RNA and protein molecules. Inhibition of these molecules can abrogate BE and GI following a radiotherapy dose, which can potentially have an application in clinical radiotherapy

Cytotoxic Activity of Enteromorpha intestinalis Extracts against Tumor Cell-Line HeLa

Two extracts (methanol and hexan) were extracted from green algae  Enteromorpha intestinalis which is isolated from Basra / Iraq, the cytotoxic activity of these extracts were evaluated by using MTT assay on HeLa cells in vitro , according to the results both extracts are effectiveness on cancer cells with the superiority of methanol extract , the IC50 of methanol extract was 79.08µg/ml and 156.3µg/ml for hexan extract . Also during the current study a number of cytotoxic compounds were isolated from both algal extracts according to the chemical analysis results by using Gc mass technique , these compounds are loliolide , palmitic acid , ethyl palmitate , phytol and squalene from methanol extract while the hexane extract contains palmitic acid , ethyl palmitate and dibutyl phthalate. Keywords : Enteromorpha intestinalis , cytotoxic activity, HeLa cell

New record of Haematotrephus nittanyense (Zeliff ,1946) Parasitized in the respiratory system of Black-winged Stilt Himantopus himantopus and White tailed Plover Chettusia leucura birds in Thi Qar Province - Iraq

Digenetic trematode, Haematotrephus nittanyense is isolated from respiratory system of Black-winged Stilt Himantopus himantopus and White-tailed Plover Chettusia Leucurus,shot in Al- Chibayish marshes , Eastern of Thi Qar province from December 2012 to November 2013 .The Prevalence in those birds are 17.03 , 2.15 % respectively . Mean intensity of infection 2.8 , 1.2 respectively.. This trematode is recorded for the first time in Iraqi birds,taxonomically described and illustrated

Three New Records of Orobanche (Orobanchaceae) to the Flora of Iraq

Three species of Orobanche L., O. alba Stephan ex Willd., O. caryophyllacea Sm. in Trans, and O. reticulata Wallr. were recorded for the first time to the flora of Iraq. O. alba is collected from Chah-Maka village, after Sheomersy from Zalan junction in Sulaimanyia District (MSU), it is parasitized on Vicia monantha. O. caryophyllacea is collected from Pira-Magron mountain (MSU), it is parasitized on Galium sp. (Rubiaceae) and O. reticulata is collected from Pera-Magron and parasitized on Vicia monantha. Key to the species, description, habitat, distribution map and photographs  were provided.

New record of Haematotrephus nittanyense (Zeliff ,1946) Parasitized in the respiratory system of Black-winged Stilt Himantopus himantopus and White tailed Plover Chettusia leucura birds in Thi Qar Province - Iraq

Digenetic trematode, Haematotrephus nittanyense is isolated from respiratory system of Black-winged Stilt Himantopus himantopus and White-tailed Plover Chettusia Leucurus,shot in Al- Chibayish marshes , Eastern of Thi Qar province from December 2012 to November 2013 .The Prevalence in those birds are 17.03 , 2.15 % respectively . Mean intensity of infection 2.8 , 1.2 respectively.. This trematode is recorded for the first time in Iraqi birds,taxonomically described and illustrated

The Impact of Vitamin D Receptor Gene Polymorphisms and Haplotypes on the susceptibility to Toxoplasmosis

Background: Toxoplasmosis is a cosmopolitan infectious disease caused by Toxoplasma gondii. Vitamin D is an immune modulator exerting its effect through a nuclear receptor called vitamin D receptor. Genetic polymorphisms in the vitamin D receptor gene could affect the activity of vitamin D and hence the individual’s susceptibility to toxoplasmosis. Objective: To evaluate the impact of four single nucleotide polymorphisms (FokI, BsmI, TaqI and ApaI) and different haplotypes of vitamin D receptor gene on the susceptibility of Iraqi women to toxoplasmosis. Patients and Methods: This case-control study involved 72 women with confirmed toxoplasmosis and 50 women as controls, DNA was extracted from blood samples and allele specific polymerase chain reaction technique was used for genotyping of the four polymorphisms using specific primers. Haplotypes and linkage disequilibrium were calculated using single nucleotide polymorphism analyzer 2.0 software. Results: Only the FokIpolymorphism had significant reverse association with toxoplasmosis in homozygote form (OR=0.140, 95%CI= 0.027-0.717, P=0.018). At allelic level, FokI F allele had significantly higher frequency in patients than controls (OR= 0.552, 95%CI=0.314-0.972, P=0.043).The frequency of two haplotypes differed significantly between patients and controls where FBAT haplotype was more frequent in patients while fta B was more frequent in controls. Moderate linkage disequilibrium correlations were found between FokI and TaqI in patients and controls. Conclusion: Allele f of FokI polymorphism and fBat haplotype in vitamin receptor gene is associated with a protective role against toxoplasmosis.

Effect of Auxin 2,4-D and NAA on Induction Callus and Adventitious Shoots and Roots regeneration from half Shoot Tips Culture of white ginger (Zingiber officinale var. Roscoe)

أجريت هذه الدراسة في مختبر زراعة الانسجة النباتية في كلية الزراعة –جامعة البصرة- العراق خلال المدة 2018-2019 بهدف دراسة تأثير تراكيز مختلفة من الاوكسين 2,4-D و  NAAفي استحثاث الكالس وتوليد الافرع والجذور العرضية من زراعة انصاف البراعم الطرفية لنبات الزنجبيل الابيضRosc.  Zingiber officinale ، تشير النتائج ان الوسط الغذائي MS المزود بالاوكسين 2,4-D بتركيز 1.0 ملغم.لتر-1 مع وجود BA بتركيز 0.5 ملغم.لتر-1 + 500 ملغم.لتر-1 PVP اعطى اعلى نسبة مئوية لاستحثاث الكالس بلغت 100% واكبر قطر ووزن طري وجاف لكتلة الكالس بلغت 2.26 سم و 1.95 و 0.12 ملغم بالتتابع، واظهرت النتائج ايضاً الى ان الوسط الغذائي MS والمزود بالاوكسين NAA بتركيز 30.0 ملغم.لتر-1 و2ip  بتركيز 3 ملغم.لتر-1 + PVP بتركيز 500 ملغم.لتر-1 قد اعطى اعلى نسبة مئوية لاستحثاث الكالس بلغت 100% في فترة قصيرة بلغت 11.33 يوم واعلى قطر ووزن طري وجاف لكتلة الكالس بلغت 2.50 سم و0.86 ملغم و0.14 ملغم بالتتابع كذلك تمت عملية توليد الجذور العرضية بيضاء اللون في جميع تراكيز NAA الا ان التركيز 10.0 ملغم.لتر-1 NAA قد اعطى اعلى معدل لعدد الجذور واطوالها بلغت 13.33 جذر/ كتلة كالس و2.83 سم بالتتابع. كذلك اوضحت النتائج عند استبدال PVP بالفحم المنشط بتركيز 500 ملغم.لتر-1 اظهرالوسط الغذائي MS المزود بالاوكسين NAA 30.0 ملغم.لتر-1 تفصص الكالس ثم ظهر نتوء اخضر صغير تميز الى فرع خضري مع ظهور جذور بيضاء اللون في نهاية فترة التحضين.This study was conducted in plant tissue culture laboratory of Agriculture College, Basrah university, Iraq during the period 2018-2019. The aims of this study were to find out the effect of different concentration of two auxin 2,4-D and NAA to induce the callus and regeneration of adventitious shoots and roots of Zingiber officinale Rosc. white via culture of half shoots tips. Results of the present study revealed that medium of MS containing 1.0 mg.l-1  2,4-D + 0.5 mg.l-1 BA + 500 mg.l-1 PVP caused primary callus formation (100%) with high diameter, fresh and dry weight of callus (2.26 cm, 1.95 mg and 0.12 mg); respectively. Results also showed that MS medium containing 30.0 mg.l-1 + 3 mg.l-1 2ip + 500 mg.l-1 PVP caused the higher primary callus formation (100%) within (11.33 days) and a higher diameter, fresh and dry weight of callus (2.50 cm , 0.86 mg and 0.14 mg); respectively. However adventitious roots were regenerated from primary callus in all concentration of NAA, whereas the medium MS containing 10.0 mg.l-1 NAA was gave the highest number and length of roots (13.33 root .callus and 2.83cm). When the activated charcoal (500 mg.l-1) was replaced by PVP in the same medium containing 30.0 mg.l-1 NAA response was found at the end of the incubation period, small green adventitious shoots and white roots were regenerated.                                                                            &nbsp

An Account of Phelipanche Pomel (Orobanchaceae) in Iraq

Full taxonomical descriptions were given to eight species of  Phelipanche Pomel (Orobanchaceae) recorded in Iraq. A key for species identification, habitat, flowering period, distribution in Iraq, general distribution and comment on species relationships were presented. Floral characteristics, photographs and maps of distribution of species were illustrated.

Preparation of some allergens from shrimp meat and evaluated of their ability to anaphylaxis's in patients suffering from shrimp allergy in Basra province ,south of Iraq

one hundred and ninety sera from subjects were tested by direct and indirect ELISA test for IgE antibodies against raw and cooked shrimp antigen respectively. The result showed that the total IgE 100>IU/ml in patients had a higher rate (98.4 %) with significant difference( p<0.01) between total IgE 100< IU/ml. distribution of shrimp antigens (raw and cooked) was determined by indirect specific IgE based ELISA, the result of this test revealed that the overall rate ( 57.3%, 74.2%) respectively of allergic patients were seropositive against all tested allergens. According to the sex of allergic patients, the female had a higher rate ( 69.2 % ) of seropositivity against raw allergens was than that of males ( 51.2 % ) with significant difference (p<0.05) between them. In concern to the effect of age on the seropositivity, the highest rate ( 60%) was observed in allergic patients (10-20 )years of age, There was no significant effect (p>0.05) for the age of allergic patients. Also, specific IgE seropositivity against the tested cooked allergens showed a higher rate in females (84.6 %) with significant difference (p<0.05) compared with males. While the effect of age on the seropositivity, other age group showed higher rate ( 83.3 %) in the 4th age group (41-50)years another age group, there was no significant difference between other age group ( p>0.05 ). Key words : Allergens,  shrimp   ,  anaphylaxis'