The effect of HIV and antiretroviral therapy on chromosomal radiosensitivity

Introduction: Antiretroviral Treatment (ART) has led to an improvement in survival of HIV infected individuals. Some of them will develop cancer during the course of their infection and will require radiation therapy. HIV positive cancer patients have presented with adverse side effects of radiotherapy and elevated chromosomal radiosensitivity. This study investigated if ART has an influence on chromosomal radiosensitivity of HIV positive individuals. Methods and Materials: Blood samples from 60 HIV positive individuals were in vitro exposed to doses of X-rays of 0, 2 and 4Gy and chromosomal radiosensitivity was assessed with the micronucleus assay. The micronucleus assay was also performed on lymphocytes of a group of non HIV-infected health care workers taking prophylactic post-exposure ART to measure the effect of these ART drugs on chromosomal radiosensitivity without HIV as a confounding factor. Results: All HIV patients (those on ART and without ART) had significantly higher radiation induced Micronuclei (MN) than healthy controls. The MN yields increased in the HIV patients taking ART compared to HIV patients not on treatment. The evaluation of chromosomal radiosensitivity of health care workers on ART revealed no effects of ART. Conclusions: HIV positive individuals show an increased chromosomal radiosensitivity. Antiretroviral treatment given to HIV positive individuals can lead to enhanced chromosomal radiosensitivity and therefore impose higher risks for radiotherapy side effects in these patients

Inactivation synergy between high energy neutrons and ⁶⁰Co gamma rays

The interaction between sublesions produced by neutrons [p(66)/Be and d(16)/Be] and ⁶⁰Co γ-rays was investigated using mammalian cells, meristematic cells and human lymphocytes. The quality of each radiation source was quantified in terms of molecular yield per unit dose absorbed in a ferrous sulphate xylenol orange solution and was found to vary inversely with the mean LET of the radiation field. Inactivation parameters determined for mammalian and meristematic cells were not significantly different following simultaneous or sequential exposures to d(16)/Be neutrons and ⁶⁰Co γ-rays. Synergistic interaction was observed to be most pronounced in a radiation mixture consisting of about one part neutrons and three parts photons and appeared to be optimal at approximately 5 Gy. This phenomenon led to dose enhancement ratios that increase with radioresistance. Multi-target parameters indicated that on a per gray basis, priming doses of p(66)/Be neutrons and ⁶⁰Co γ-rays induce comparative levels of sublethal damage. However, non-parametric analysis of the survival data showed that mammalian cells regard a priming dose of neutrons as somewhat less effective than an iso-effective photon dose. A greater measure of synergy was observed between photons and priming doses of neutrons with less build-up. This is however mainly due to higher levels of biological damage induced with a more potent configuration of secondary charged particles. Interaction factors compared at levels of iso-effect tend to be smaller when the LET of the priming dose was increased. Split-doses of neutrons in the absence of build-up resulted in "negative" repair. The validity of proposed biophysical models was tested using meristematic cells, as the response of these cells show an apparent absence of intertrack damage. Contrary to expectations, synergistic interaction was observed for both growth delay measurements and micronuclear formations. Chromosome aberrations showed synergy between neutron and photon damage in human lymphocytes, as predicted by interaction functi ons. However, the synergistic interaction noted with micronuclear formation in binucleate cells was at variance with predictions based on biophysical models

Chromosomal radiosensitivity of human immunodeficiency virus positive/negative cervical cancer patients in South Africa

Cervical cancer is the second most common cancer amongst South African women and is the leading cause of cancer-associated mortality in this region. Several international studies on radiation-induced DNA damage in lymphocytes of cervical cancer patients have remained inconclusive. Despite the high incidence of cervical cancer in South Africa, and the extensive use of radiotherapy to treat it, the chromosomal radiosensitivity of South African cervical cancer patients has not been studied to date. Since a high number of these patients are human immunodeficiency virus (HIV)-positive, the effect of HIV infection on chromosomal radiosensitivity was also investigated. Blood samples from 35 cervical cancer patients (20 HIV-negative and 15 HIV-positive) and 20 healthy controls were exposed to X-rays at doses of 6 MV of 2 and 4 Gy in vitro. Chromosomal radiosensitivity was assessed using the micronucleus (MN) assay. MN scores were obtained using the Metafer 4 platform, an automated microscopic system. Three scoring methods of the MNScore module of Metafer were applied and compared. Cervical cancer patients had higher MN values than healthy controls, with HIV-positive patients having the highest MN values. Differences between groups were significant when using a scoring method that corrects for false positive and false negative MN. The present study suggested increased chromosomal radiosensitivity in HIV-positive South African cervical cancer patients

The radiosensitizing effect of Ku70/80 knockdown in MCF10A cells irradiated with X-rays and p(66)+Be(40) neutrons

Background: A better understanding of the underlying mechanisms of DNA repair after low-and high-LET radiations represents a research priority aimed at improving the outcome of clinical radiotherapy. To date however, our knowledge regarding the importance of DNA DSB repair proteins and mechanisms in the response of human cells to high-LET radiation, is far from being complete. Methods: We investigated the radiosensitizing effect after interfering with the DNA repair capacity in a human mammary epithelial cell line (MCF10A) by lentiviral-mediated RNA interference (RNAi) of the Ku70 protein, a key-element of the non-homologous end-joining (NHEJ) pathway. Following irradiation of control and Ku-deficient cell lines with either 6 MV X-rays or p(66)+Be(40) neutrons, cellular radiosensitivity testing was performed using a crystal violet cell proliferation assay. Chromosomal radiosensitivity was evaluated using the micronucleus (MN) assay. Results: RNAi of Ku70 caused downregulation of both the Ku70 and the Ku80 proteins. This downregulation sensitized cells to both X-rays and neutrons. Comparable dose modifying factors (DMFs) for X-rays and neutrons of 1.62 and 1.52 respectively were obtained with the cell proliferation assay, which points to the similar involvement of the Ku heterodimer in the cellular response to both types of radiation beams. After using the MN assay to evaluate chromosomal radiosensitivity, the obtained DMFs for X-ray doses of 2 and 4 Gy were 2.95 and 2.66 respectively. After neutron irradiation, the DMFs for doses of 1 and 2 Gy were 3.36 and 2.82 respectively. The fact that DMFs are in the same range for X-rays and neutrons confirms a similar importance of the NHEJ pathway and the Ku heterodimer for repairing DNA damage induced by both X-rays and p(66)+Be(40) neutrons. Conclusions: Interfering with the NHEJ pathway enhanced the radiosensitivity of human MCF10A cells to low-LET Xrays and high-LET neutrons, pointing to the importance of the Ku heterodimer for repairing damage induced by both types of radiation. Further research using other high-LET radiation sources is however needed to unravel the involvement of DNA double strand break repair pathways and proteins in the cellular response of human cells to high-LET radiation

Radiosensitization Effect of Gold Nanoparticles in Proton Therapy

The number of proton therapy facilities and the clinical usage of high energy proton beams for cancer treatment has substantially increased over the last decade. This is mainly due to the superior dose distribution of proton beams resulting in a reduction of side effects and a lower integral dose compared to conventional X-ray radiotherapy. More recently, the usage of metallic nanoparticles as radiosensitizers to enhance radiotherapy is receiving growing attention. While this strategy was originally intended for X-ray radiotherapy, there is currently a small number of experimental studies indicating promising results for proton therapy. However, most of these studies used low proton energies, which are less applicable to clinical practice; and very small gold nanoparticles (AuNPs). Therefore, this proof of principle study evaluates the radiosensitization effect of larger AuNPs in combination with a 200 MeV proton beam. CHO-K1 cells were exposed to a concentration of 10 μg/ml of 50 nm AuNPs for 4 hours before irradiation with a clinical proton beam at NRF iThemba LABS. AuNP internalization was confirmed by inductively coupled mass spectrometry and transmission electron microscopy, showing a random distribution of AuNPs throughout the cytoplasm of the cells and even some close localization to the nuclear membrane. The combined exposure to AuNPs and protons resulted in an increase in cell killing, which was 27.1% at 2 Gy and 43.8% at 6 Gy, compared to proton irradiation alone, illustrating the radiosensitizing potential of AuNPs. Additionally, cells were irradiated at different positions along the proton depth-dose curve to investigate the LET-dependence of AuNP radiosensitization. An increase in cytogenetic damage was observed at all depths for the combined treatment compared to protons alone, but no incremental increase with LET could be determined. In conclusion, this study confirms the potential of 50 nm AuNPs to increase the therapeutic efficacy of proton therapy

RENEB intercomparison exercises analyzing micronuclei (Cytokinesis-block Micronucleus Assay)

Purpose: In the framework of the ‘Realizing the European Network of Biodosimetry’ (RENEB) project, two intercomparison exercises were conducted to assess the suitability of an optimized version of the cytokinesis-block micronucleus assay, and to evaluate the capacity of a large laboratory network performing biodosimetry for radiation emergency triages. Twelve European institutions participated in the first exercise, and four non-RENEB labs were added in the second one. Materials and methods: Irradiated blood samples were shipped to participating labs, whose task was to culture these samples and provide a blind dose estimate. Micronucleus analysis was performed by automated, semi-automated and manual procedures. Results: The dose estimates provided by network laboratories were in good agreement with true administered doses. The most accurate estimates were reported for low dose points (== 2.7 Gy) a larger variation in estimates was observed, though in the second exercise the number of acceptable estimates increased satisfactorily. Higher accuracy was achieved with the semi-automated method. Conclusion: The results of the two exercises performed by our network demonstrate that the micronucleus assay is a useful tool for large-scale radiation emergencies, and can be successfully implemented within a large network of laboratories

RENEB intercomparisons applying the conventional Dicentric Chromosome Assay (DCA)

Purpose: Two quality controlled inter-laboratory exercises were organized within the EU project ‘Realizing the European Network of Biodosimetry (RENEB)’ to further optimize the dicentric chromosome assay (DCA) and to identify needs for training and harmonization activities within the RENEB network. Materials and methods: The general study design included blood shipment, sample processing, analysis of chromosome aberrations and radiation dose assessment. After manual scoring of dicentric chromosomes in different cell numbers dose estimations and corresponding 95% confidence intervals were submitted by the participants. Results: The shipment of blood samples to the partners in the European Community (EU) were performed successfully. Outside the EU unacceptable delays occurred. The results of the dose estimation demonstrate a very successful classification of the blood samples in medically relevant groups. In comparison to the 1st exercise the 2nd intercomparison showed an improvement in the accuracy of dose estimations especially for the high dose point. Conclusions: In case of a large-scale radiological incident, the pooling of ressources by networks can enhance the rapid classification of individuals in medically relevant treatment groups based on the DCA. The performance of the RENEB network as a whole has clearly benefited from harmonization processes and specific training activities for the network partners

Die toepassing van die beginsels en metodes van moderne bemarkingsbestuur in die bemarking van dienste met spesiale verwysing na die goederedienste van die Suid- Afrikaanse Spoorweë

M.Com.This study originated from the thought that the principles and practices of modern marketing management can be used more effectively in the South African Railways as one of the largest public organisations in the world. The objectives of the study were an investigation into the principles and practices of modern marketing management in the service industry in general and the extent to which these principles and practices are applied in the marketing of goods services by the South African Railways. The study included a survey of the literature on the marketing of services, including some railway organisations in other countries as well as an in depth investigation of the present approach to and practice of marketing management in the South African Railways

Assessment of changes in photosystem II structure and function as affected by water deficit in Amaranthus hypochondriacus L. and Amaranthus hybridus L

The present study describes the behaviour of photosystem II (PSII) in Amaranthus hypochondriacus and Amaranthus hybridus under water stress conditions, assessed by the analyses of the polyphasic rise in chlorophyll a fluorescence (O-J-I-P). We determined the adaptive behaviour in relation to the regulation of the different functional and structural parameters of PSII, which was a direct and rapid response due to changes in soil water status indicated by a decrease in leaf water potential and relative water content. It allows for the identification of specific key or limiting chlorophyll fluorescence parameters which could be used to identify traits conveying tolerance. For the above partial processes of PSII function studied, it seems that A. hybridus remained the more stable upon water stress (after 17 days of withholding water), concerning the specific energy fluxes of absorption/reaction centre (ABS/RC) apparent (antenna size) and trapping/reaction centre (TR/RC) (maximum trapping flux), as well as the density of the reaction centres/cross section (RC/CS) and the phenomenological trapping flux/cross section (TRo/CS). It was clear that amaranth adjusts the non-photochemical (kn) deactivation constant of PSII and to a less extend also the photochemical (kp) deactivation constant by means of photoregulation, which forms the basis of the quenching of chlorophyll a fluorescence. Although drought stress caused the deactivation of RCs leading to a decrease in the density of active RCs, the plants compensated by increasing the efficiency of the conversion of trapped excitation energy to electron transport beyond QA (efficiency of exciton trapping/reaction centre: ETo/TRo). Subsequent damage to PSII might be the reason for the slow, or lack of recovery, for most of the parameters measured.http://dx.doi.org/10.1016/j.plaphy.2011.05.00