Multiple myeloma results from the clonal proliferation of abnormal plasma cells (PCs) in the bone marrow (BM). In this study, the cell surface expression markers (CD) on atypical PCs (detected by multiparametric flow cytometry (MFC)) were correlated with copy number alterations (CNAs) in the genome (detected by multiplex ligation-dependent probe amplification (MLPA)) to assess their impact on prognosis in newly diagnosed MM patients. Statistically significant results were obtained when different stages of PC maturation (classified based on CD19 and CD81 expression) were associated with CD117 expression and identified CNAs. In the intermediately differentiated PC group (CD19(−) CD81(+)), patients who didn’t express CD117 had a lower median progression free survival (PFS) (p = 0.024). Moreover, within this group, patients with less than three adverse CNAs, which harbor CD117, had a better outcome with a PFS of more than 48 months compared with 19 months (p = 0.008). Considering all the results, our study suggested the need to integrate both the CD markers and copy number alterations to evaluate the prognosis of newly diagnosed multiple myeloma patients.
Over the past decade, cold atmospheric plasma (CAP) has undergone extensive research as a promising therapeutic approach in oncology, with different treatment methods and exposure configurations being investigated and resulting in various biological effects, most of them after long exposure or treatment durations. This study aimed to evaluate the potential of a custom-made CAP generation source to produce plasma-activated medium (PAM) with cytotoxic effects and subsequently to establish the optimal exposure and treatment parameters. The exposure’s electrical parameters, as well as pH and content of PAM were analyzed. The cytotoxic potential and optimal parameters of the treatment were established by evaluating the viability of human osteosarcoma cells (HOS cell line) and human osteoblasts (HOB cell line) treated with PAM under different conditions. Our results showed that indirect treatment with CAP presents selective dose-dependent cytotoxic effects, while the cell viability decrease was not found to be correlated with the PAM acidification due to CAP exposure. The Griess assay revealed very high and long-term stable concentrations of in PAM. Overall, this study presents a simple and faster method of producing PAM treatment with cytotoxic effects on HOS cells, by using a custom-built CAP source.
We evaluated, by the flow cytometry technique, the viability of two tumor cell lines: colorectal carcinoma (HT-29) and human A549 arcinoma incubated with the cytotoxic peptide LL-37. The results obtained for the two cell lines HT-29 and A549 are significantly different under the action of cathelicidin LL-37. At high concentrations of 20mM, cellular apoptosis was over 30% higher for colorectal adenocarcinoma line compared to that by peptide exposure. Apoptosis was also significant in low-concentration (4uM) catechidine-labeled lung cancer cells for 48 h. Also, optimization of primers was sought to evaluate gene expression for, Bcl2, IL6, IL8. Determination of gene expression for these molecular targets under the action of the cytotoxic peptide was performed in order to evaluate the immune response of tumor cells. For this purpose, the genetic material (RNA) was extracted from cell cultures and reversed in cDNA, which was subsequently amplified by the qRT-PCR technique. Evaluation of tumor cell metabolism was done by determining the gene expression for Bcl2, IL6, IL8 by the action of the cytotoxic peptides used. The cytotoxic effect of cathelicidin LL-37 for the two cell lines HT29 and A549 is supported by the decrease in IL-6 and IL-8 gene expression. The increase in Bcl2 expression for cells exposed to the action of the peptide is explained by the increase in anti-apoptotic protein synthesis that explains the fight of tumor cells for survival and proliferation.
Many studies have highlighted the anti-tumor properties of some natural peptides known to have antimicrobial virtues. In this study, we evaluated the tumoricidal potential of dermaseptin, defensin, cecropin A and B on tumor cell lines: M14K (human mesothelioma). The viability study was performed using PE V Annexin and 7-AAD (7-amino-actinomycin D) (BD Pharmingen). This test was used to detect and measure apoptosis by flow cytometry technique. The experimental results of our study revealed that the cytotoxic effects of the four peptides depend on their concentration. In this in vitro experimental study, we found that the cytotoxic effect of the four cytotoxic peptides used depended on their concentration in the tumor cell culture medium, being significant at concentrations of 120mM and maintained at concentrations of 60�M. At 30�M concentrations these tumoricidal effects were insignificant. Of all the studied peptides, dermaseptin has the most powerful effect and the weakest effect b - defensin - 1.
Background/Objectives: KDIGO supports molecular testing in selected autosomal dominant polycystic kidney disease (ADPKD) scenarios and targeted next-generation sequencing panels when broader evaluation is warranted; however, gene inclusion alone establishes neither analytical completeness nor clinical reportability. Methods: We designed and evaluated a 28-gene ADPKD-spectrum hybrid-capture panel in a 16-sample, two-configuration pilot using 694 analytical target intervals per sample. Multiplex ligation-dependent probe amplification assessed dosage in suspected PKD1-related disease or TSC2/PKD1 contiguous-gene deletion, and whole-exome sequencing selectively supported short-read reidentification. Results: In the primary eight-sample dataset, 4592/5552 observations (82.7%) had interval mean depth ≥20×, 4256/5552 (76.7%) had interval minimum depth ≥20×, and 89.6% of interval-record bases reached ≥10×. Nine genes (FLCN, GANAB, HNF1B, PRKCSH, REN, SEC61A1, TSC2, UMOD and VHL) met mean depth ≥20× throughout; six also met the minimum-depth criterion. For PKD1, 45/46 intervals met both criteria in all samples, but exon 1 remained undercovered, and nominal depth in duplicated sequence did not establish authentic-locus callability. Signals were observed at all four positive-comparator loci, although two lacked sufficient support for independent reporting. Seven of nine panel-first cases yielded observations: one HNF1B and one unique-locus PKD1 finding were supported, whereas four duplicated-sequence PKD1 findings remained confirmation-dependent. Conclusions: Responsible implementation requires explicit analytical boundaries and staged complementary testing.
Background: Technology allows us to predict a histopathological diagnosis, but the high costs prevent the large-scale use of these possibilities. The current liberal indication for surgery in benign thyroid conditions led to a rising frequency of incidental thyroid carcinoma, especially low-risk papillary micro-carcinomas. Methods: We selected a cohort of 148 patients with thyroid nodules by ultrasound characteristics and investigated them by fine needle aspiration cytology (FNAC)and prospective BRAF collection for 70 patients. Also, we selected 44 patients with thyroid nodules using semi-quantitative functional imaging with an oncological, 99mTc-methoxy-isobutyl-isonitrile (99mTc-MIBI) radiotracer. Results: Following a correlation with final histopathological reports in patients who underwent thyroidectomy, we introduced the results in a machine learning program (AI) in order to obtain a pattern. For semi-quantitative functional visual pattern imaging, we found a sensitivity of 33%, a specificity of 66.67%, an accuracy of 60% and a negative predicting value (NPV) of 88.6%. For the wash-out index (WOind), we found a sensitivity of 57.14%, a specificity of 50%, an accuracy of 70% and an NPV of 90.06%.The results of BRAF in FNAC included 87.50% sensitivity, 75.00% specificity, 83.33% accuracy, 75.00% NPV and 87.50% PPV. The prevalence of malignancy in our small cohort was 11.4%. Conclusions: We intend to continue combining preoperative investigations such as molecular detection in FNAC, 99mTc-MIBI scanning and AI training with the obtained results on a larger cohort. The combination of these investigations may generate an efficient and cost-effective diagnostic tool, but confirmation of the results on a larger scale is necessary.