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

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.

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.

Antibody Response of Broiler Chickens against Eight Commercial Infectious Bursal Disease Live Vaccines tested by ELISA

This study was designed to investigate the antibody response of broiler chickens against eight commercial IBD live vaccines. A total of 460 one- day Ross broiler chicks were divided to 9 groups, eight groups were vaccinated with IBD live vaccines and the last one was served as control. Four groups were vaccinated with intermediated vaccines, whereas the other fourth were given intermediated plus vaccines. All vaccinated groups were administrated at 14th day of age via drinking water route. Maternal derived antibody (MDA) and post- vaccination antibody response were tested by ELISA. Blood samples were collected at one-day old and at 21st, 28th and 35th day of age post- vaccination. Indirect ELISA test revealed that the mean of maternal derived antibody was 4852±745. Significant differences (P<0.05) among means of antibody titers of all vaccinated groups were found at 21st ,28th and 35th day of age compared with that of control group. The results also showed that groups which were vaccinated with intermediated plus vaccines (E and H vaccines) exhibited high level of antibody especially groups 5 and 8 than those which vaccinated with intermediate vaccines. In conclusion, Intermediate plus vaccines induced higher antibody titers than other vaccines, although some intermediate vaccines induced similar titers of antibody E and H vaccines which were administered to groups 5 and 8 respectively induced better antibody titers

Influence of antibiotics treatment on hematological aspect in chicken

Abstract: Two experiments were described in which sixty one-day old (IPA) chicks were distributed into four equal groups, three for treatment &amp; the other for control. Ampicillin, Enrofloxacine or Amoxicillin were used for five consecutive days via drinking water for treatment for at the first and twenty second day of life. Five birds from each group were slaughtered on 6 and 28 days post treatment and hematological examinationsperformed on blood samples collected just prior to slaughter. Changes in PCV, HB, RBC, WBC, MCV, MCH and MCHC were observed. The investigations demonstrated a fall in hematological values in body chicks, Whereas in older ages these values were analogous with normal ranges. The changes were not substantiate the presence of anemia. They might be an indications of an incidental haemodilutionin the hematological observed in this study

An account of the genus Cistanche (Orobanchaceae) in Iraq and taxonomic considerations in the Middle East

Species limits in the genus Cistanche are poorly understood, despite the plants’ long history of use in traditional herbal medicine and food across their range. Here we present a taxonomic account for the genus Cistanche in Iraq, where several taxa have been reported, most of them doubtfully. Using herbarium specimens, images of living material, and taxonomic literature, we found evidence of only one species occurring with certainty in Iraq: Cistanche tubulosa. We found no evidence for the occurrence of other Cistanche species in Iraq, including a putative new entity reported for the region. Our work highlights inconsistencies in the literature, and underscores the importance of examining multiple stable characters for delimiting species in the genus Cistanche