Ghazi Alsbeih

The phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) plays a critical role in cell growth and survival. PIK3CA somatic mutations are linked to the PIK3CA-related overgrowth spectrum (PROS) syndrome. Cells from those patients appeared to be associated with a moderate but significant radiosensitivity. Mutations or amplifications in this gene are common in breast, colorectal, and lung cancers. Alterations in the PIK3CA gene, including amplification and mutation, are common in cancer, but their influence on radiotherapy is not yet fully understood. This report reveals a potential association between PIK3CA gene modifications and radiosensitivity (p < 0.05) deducted from 8 established colorectal cancer cell lines (HT-29, DLD-1, HCT-116, SW480, HCT-15, Colo-320, and LoVo). Meanwhile gene amplification (> 2) seems to be linked to increased radiation sensitivity, mutations appear to be associated with increased radioresistance in colorectal cancer cells. Leveraging this relationship, PIK3CA amplification and mutations could act as biomarker to pinpoint patients who might benefit from more personalized radiotherapy regimens. However, this association is preliminary and hypothesis-generating in view of the limited number of cases. Further studies are needed to confirm this conclusion. By uncovering the distinct mechanistic effects of these PIK3CA alterations on radiosensitivity phenotype in both normal and cancerous cells, researchers can lay the groundwork for tailored radiotherapy strategies in colorectal cancer. This insight could enhance treatment effectiveness while reducing side effects, ultimately leading to improved patient outcomes.

The search for NF-κB blockers to optimize antioxidants and counteract inflammation and carcinogenesis has identified several promising natural compounds, encompassing curcumin. Nonetheless, despite the pleiotropic health attributes of curcumin and established safety, its in vivo efficacy is limited by its poor pharmacokinetic properties, primarily due to its low bioavailability because of its low free serum concentrations and short half-life. To overcome these restrictions, we investigated the efficacy of the synthetic curcumin analog PAC in breast cancer cells. Additionally, MCF-7 and MDA-MB-231 cell lines were employed to explore the anticancer effects of PAC by assessing cell viability, apoptosis, clonogenic survival, and the expression of NF-κB, p53, Bcl-2, and Bax. Our results validate that PAC effectively inhibits cell proliferation and induces apoptosis in a dose-dependent manner. PAC also significantly enhances the radiosensitivity of breast cancer cells, indicating a potential synergistic effect with radiation therapy. Mechanistically, PAC exerts anticancer effects by inhibiting NF-κB signaling and modulating apoptotic genes like p53, Bax, and Bcl-2. These findings highlight its potential as a breast cancer therapeutic, enhancing existing treatments.

Venom peptides are promising agents in the development of unconventional anticancer therapeutic agents. This study explored the potential of Pilosulin-3, a recombinant peptide from the venom of the Australian jack jumper ant “Myrmecia pilosula”, as a cytotoxic and radiosensitizing agent in MCF-7 and MDA-MB-231 breast cancer (BC) cell lines. Pilosulin-3’s cytotoxicity was evaluated across a wide range of concentrations using a proliferation assay. Cell cycle progression and apoptosis were examined at the inhibitory concentration 25% (IC25) and IC50 of Pilosulin-3, both with and without a 4Gy X-ray irradiation dose. Radiosensitivity was assessed at IC25 using the clonogenic survival assay. The study revealed that Pilosulin-3 exerted a concentration-dependent cytotoxic effect, with IC25 and IC50 values of 0.01 and 0.5 µM, respectively. In silico screening indicated high selectivity of Pilosulin-3 peptide, which was predicted to be the most likely anticancer agent (PROB = 0.997) with low hemolytic activity (PROP = 0.176). Although Pilosulin-3 exhibited a significant (p < 0.05) G2/M cell cycle arrest in combination with radiation, there was no discernible effect on apoptosis induction or cell survival following irradiation. In conclusion, Pilosulin-3 proved to be cytotoxic to BC cells and induced a cytostatic effect (G2/M arrest) when combined with radiation. However, it did not enhance the efficacy of cell killing by irradiation. While it holds potential as a cytotoxic agent in breast cancer treatment, its application as a radiosensitizer does not find support in these results.

Introduction: The search for biomarkers to predict radiosensitivity is important not only to individualize radiotherapy of cancer patients but also to forecast radiation exposure risks. The aim of this study was to devise a machine-learning method to stratify radiosensitivity and to investigate its association with genome-wide copy number variations (CNVs) as markers of sensitivity to ionizing radiation.Methods: We used the Affymetrix CytoScan HD microarrays to survey common CNVs in 129 fibroblast cell strains. Radiosensitivity was measured by the surviving fraction at 2 Gy (SF2). We applied a dynamic programming (DP) algorithm to create a piecewise (segmented) multivariate linear regression model predicting SF2 and to identify SF2 segment-related distinctive CNVs.Results: SF2 ranged between 0.1384 and 0.4860 (mean=0.3273 The DP algorithm provided optimal segmentation by defining batches of radio-sensitive (RS), normally-sensitive (NS), and radio-resistant (RR) responders. The weighted mean relative errors (MRE) decreased with increasing the segments' number. The borders of the utmost segments have stabilized after partitioning SF2 into 5 subranges.Discussion: The 5-segment model associated C-3SFBP marker with the most-RS and C-7IUVU marker with the most-RR cell strains. Both markers were mapped to gene regions (MCC and SLC1A6, respectively). In addition, C-3SFBP marker is also located in enhancer and multiple binding motifs. Moreover, for most CNVs significantly correlated with SF2, the radiosensitivity increased with the copy-number decrease.In conclusion, the DP-based piecewise multivariate linear regression method helps narrow the set of CNV markers from the whole radiosensitivity range to the smaller intervals of interest. Notably, SF2 partitioning not only improves the SF2 estimation but also provides distinctive markers. Ultimately, segment-related markers can be used, potentially with tissues’ specific factors or other clinical data, to identify radiotherapy patients who are most RS and require reduced doses to avoid complications and the most RR eligible for dose escalation to improve outcomes.

To cope with the shortage of filtering facepiece respirators (FFRs) during the coronavirus (COVID-19) pandemic, healthcare institutions were forced to reuse FFRs after applying different decontamination methods including gamma-irradiation (GIR). The aim of this study was to evaluate the effect of GIR on the filtration efficiency (FE) of FFRs and on SARS-CoV-2 detection. The FE of 2 FFRs types (KN95 and N95-3 M masks) was assessed at different particle sizes (0.3–5 µm) following GIR (0–15 kGy) delivered at either typical (1.65 kGy/h) or low (0.5088 kGy/h) dose rates. The detection of two SARS-CoV-2 RNA genes (E and RdRp4) following GIR (0–50 kGy) was carried out using RT-qPCR assay. Both masks showed an overall significant (P < 0.001) reduction in FE with increased GIR doses. No significant differences were observed between GIR dose rates on FE. The GIR exhibited significant increases (P ≤ 0.001) in the cycle threshold values (ΔCt) of both genes, with no detection following high doses. In conclusion, complete degradation of SARS-CoV-2 RNA can be achieved by high GIR (≥ 30 kGy), suggesting its potential use in FFRs decontamination. However, GIR exhibited adverse effects on FE in dose- and particle size-dependent manners, rendering its use to decontaminate FFRs debatable.

To cope with the shortage of filtering facepiece respirators (FFRs) caused by the coronavirus disease (COVID-19), healthcare institutions have been forced to reuse FFRs using different decontamination methods, including vapor hydrogen peroxide (VHP). However, most healthcare institutions still struggle with evaluating the effect of VHP on filtration efficiency (FE) of the decontaminated FFRs. We developed a low-cost in-house FE assessment using a novel 3D-printed air duct. Furthermore, we assessed the FE of seven types of FFRs. Following 10 VHP cycles, we evaluated the FE of KN95 and 3M-N95 masks. The 3M-N95 and Benehal-N95 masks showed significant lower FE (80.4–91.8%) at fine particle sizes (0.3–1 µm) compared to other FFRs (FE ≥ 98.1%, p < 0.05). Following 10 VHP cycles, the FE of KN95 masks was almost stable (FE stability > 99.1%) for all particle sizes, while 3M-N95 masks were stable only at 2 and 5 µm (FE stability > 98.0%). Statistically, FE stability of 3M-N95 masks at 0.3, 0.5, and 0.7 µm was significantly lower (p ≤ 0.006) than 2 and 5 µm. The in-house FE assessment may be used as an emergency procedure to validate the decontaminated FFRs, as well as a screening option for production control of FFRs. Following VHP cycles, both masks showed high stability at 5 µm, the size of the most suspected droplets implicated in COVID-19 transmission.

We tested the hypothesis that differences in DNA double-strand break (DSB) repair fidelity underlies differences in individual radiosensitivity and, consequently, normal tissue reactions to radiotherapy. Fibroblast cultures derived from a radio-sensitive (RS) breast cancer patient with grade 3 adverse reactions to radiotherapy were compared with normal control (NC) and hyper-radiosensitive ataxia-telangiectasia mutated (ATM) cells. DSB repair and repair fidelity were studied by Southern blotting and hybridization to Alu repetitive sequence and to a specific 3.2-Mbp NotI restriction fragment on chromosome 21, respectively. Results for DNA repair kinetics using the NotI fidelity assay showed significant differences (P < 0.001) with higher levels of misrepaired (misrejoined and unrejoined) DSBs in RS and ATM compared with NC. At 24-h postradiation, the relative fractions of misrepaired DSBs were 10.64, 23.08, and 44.70% for NC, RS, and ATM, respectively. The Alu assay showed significant (P < 0.05) differences in unrepaired DSBs only between the ATM and both NC and RS at the time points of 12 and 24 h. At 24 h, the relative percentages of DSBs unrepaired were 1.33, 3.43, and 12.13% for NC, RS, and ATM, respectively. The comparison between the two assays indicated an average of 5-fold higher fractions of misrepaired (NotI assay) than unrepaired (Alu assay) DSBs. In conclusion, this patient with increased radiotoxicity displayed more prominent misrepaired than unrepaired DSBs, suggesting that DNA repair fidelity is a potential marker for the adverse reactions to radiotherapy. More studies are required to confirm these results and further develop DSB repair fidelity as a hallmark biomarker for interindividual differences in radiosensitivity.

In cases of nuclear and radiological accidents, public health and emergency response need to assess the magnitude of radiation exposure regardless of whether they arise from disaster, negligence, or deliberate act. Here we report the establishment of a national reference dose–response calibration curve (DRCC) for dicentric chromosome (DC), prerequisite to assess radiation doses received in accidental exposures. Peripheral blood samples were collected from 10 volunteers (aged 20–40 years, median = 29 years) of both sexes (three females and seven males). Blood samples, cytogenetic preparation, and analysis followed the International Atomic Energy Agency EPR-Biodosimetry 2011 report. Irradiations were performed using 320 kVp X-rays. Metafer system was used for automated and assisted (elimination of false-positives and inclusion of true-positives) metaphases findings and DC scoring. DC yields were fit to a linear–quadratic model. Results of the assisted DRCC showed some variations among individuals that were not statistically significant (homogeneity test, P = 0.66). There was no effect of age or sex (P > 0.05). To obtain representative national DRCC, data of all volunteers were pooled together and analyzed. The fitted parameters of the radiation-induced DC curve were as follows: Y = 0.0020 (±0.0002) + 0.0369 (±0.0019) ^*D + 0.0689 (±0.0009) ^*D². The high significance of the fitted coefficients (z-test, P < 0.0001), along with the close to 1.0 p-value of the Poisson-based goodness of fit (χ² = 3.51, degrees of freedom = 7, P = 0.83), indicated excellent fitting with no trend toward lack of fit. The curve was in the middle range of DRCCs published in other populations. The automated DRCC over and under estimated DCs at low (2 Gy) doses, respectively, with a significant lack of goodness of fit (P < 0.0001). In conclusion, we have established the reference DRCC for DCs induced by 320 kVp X-rays. There was no effect of age or sex in this cohort of 10 young adults. Although the calibration curve obtained by the automated (unsupervised) scoring misrepresented dicentric yields at low and high doses, it can potentially be useful for triage mode to segregate between false-positive and near 2-Gy exposures from seriously irradiated individuals who require hospitalization.

Flow Cytometry-Based Gamma-H2AX Assay for Radiation Biodosimetry protocol (method) by Ghazi Alsbeih