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Osteosarcoma (OS), a malignancy primarily affecting children and adolescents, is the most frequently encountered malignant, non-hematologic, bone tumor. Despite the gradual improvement of survival rates, the management of this disease remains problematic due to challenges such metastasis development, its heterogeneous characteristics, and resistance to cytostatic drugs. Cold atmospheric plasma (CAP), a partially ionized gas operating at near room temperature, which is comprised of free carriers, excited or neutral molecules, and active radicals capable of initiating diverse physical phenomena and chemical reactions represent a new and innovative potential solution in cancer therapy. The aim of this study was to evaluate the capacity of CAP produced by our custom-build plasma source to induce cytotoxic effects to HOS cells and to analyze post-treatment modulations of cdk1, tnf α and tp53 genes expression. Direct and indirect CAP treatments effectiveness on HOS cells were evaluated by MTT assay and the regulation of interest genes expression were carried out by RT-qPCR analysis. Cell viability analysis revealed a strong cytotoxic effect of direct CAP treatment on HOS cells, while the indirect treatment resulted in a slight decrease of cells viability. Direct CAP treatment modulates the expression of all analyzed genes, both at 2- and 24-hours posttreatment. In conclusion, direct CAP treatment produced by our custom-build plasma source have cytotoxic effects on HOS cells up to 24 hours post-treatment. Furthermore, direct CAP treatment induces cell cycle arrest of HOS cells, and the CAP-induced cell death is independent of tp53 gene. [ABSTRACT FROM AUTHOR]

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 NO 2 − 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 NO 2 − 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. [ABSTRACT FROM AUTHOR]

Dizabilitatea intelectuală (DI) este o afecțiune frecventă, cu consecințe majore pentru individ, familie și societate. Datorită heterogenității sale clinice și genetice, în aproximativ 50% din cazuri etiologia bolii nu poate fi stabilită. Scopul acestui studiu a fost evaluarea capacității de stabilire a diagnosticului etiologic la 369 pacienți cu DI sindromic și rezultat normal sau incert la cariotip folosind o combinație de kituri MLPA. Toţi pacienţii au fost investigaţi prin metoda MLPA, folosind fie kiturile SALSA MLPA P064 sau P096, dacă fenotipul a fost sugestiv pentru un sindrom cu microdeleţie (subgrupul A - 186 pacienți), fie kiturile subtelomerice P036 și P070, dacă fenotipul nu a fost sugestiv pentru un sindrom cu microdeleţie sau rezultatul la cariotipul standard a fost incert (subgrupul B - 183 pacienți). Rezultatele anormale detectate de aceste kituri au fost caracterizate folosind kiturile MLPA corespunzătoare de urmărire (Telomere Follow-up set, P029-A1, P250-B2, ME028-B1). În subgrupul A am identificat 25 de pacienți cu microdeleții (13,4%). Folosind kiturile de screening subtelomeric și de urmărire la subgrupul B am detectat rearanjări criptice în 7,5% din cazuri și am identificat originea materialului suplimentar observat la cariotipul standard la 10 din 11 pacienți. Sumarizând datele obţinute din cele două loturi, folosirea combinată a seturilor MLPA a dus la stabilirea diagnosticului la 10,6% (38/358) dintre pacienții cu cariotip normal. Folosirea seturilor MLPA de urmărire a permis atât confirmarea prezenţei anomaliei, cât şi determinarea dimensiunii ei, ceea ce a facilitat interpretarea semnificaţiei clinice a rearanjărilor. Pentru laboratoarele care nu au acces la tehnologiile bazate pe microarray, folosirea mai multor kituri MLPA reprezintă o strategie eficientă pentru stabilirea diagnosticului etiologic la pacienţii cu DI.

Introduction: The intestinal microbiota is vital to human health, and has a profound influence on several biological processes including inflammation and pathogen resistance. Antibiotic intake greatly impacts bacterial diversity, can increase antibiotic resistance and impair the equilibrium between bacterial species. The key to grasping post-antibiotic effects on the gut microbiota rests on the implementation of a suitable procedure to isolate microbial DNA and a meticulous consideration of experimental sequencing artefacts. Methods: We herein report the bacterial community dynamics of a cohort of 128 surgical oncology patients before and after the intravenous administration of cefuroxime, an antibiotic routinely used in surgical antibioprophylaxis with proven efficiency against both gram-positive and gram-negative bacteria. In our study, we analyzed patient fecal samples collected through rectal examination before and 7 days post cefuroxime treatment by employing a high-throughput sequencing assay which targets the V3-V4 region of the 16S rRNA gene. A first challenge in applying the study design was to extract an appropriate amount of DNA characteristic to the sampled microbiota, which implied the use of both mechanical (ceramic beads) and chemical (proteinase K, lysozyme and lysostaphin) lysis. Results: Gut microbiota richness and composition was significantly different between the two groups, but most differences were determined by additional perioperative procedures, rather than antibioprophylaxis. Intestinal microbiota composition was not significantly changed one week post cefuroxime treatment when compared to pre-treatment condition for patients without mechanical bowel preparation, but some loss in taxonomic variety could be observed. Discussion: Taken together, cefuroxime does not promote short-term dysbiosis in surgical patients without any additional perioperative procedures. [ABSTRACT FROM AUTHOR]

Multiple myeloma (MM) is characterized by the uncontrolled proliferation of an atypical plasma cell clone. The main factor preventing the characterization of plasma cells is their low rate in the analyzed samples. Through this study we tried to highlight the necessity of enrichment of MM samples to be analyzed by MLPA and SNParray. Samples from 5 patients diagnosed with MM were investigated and 2 of them were enriched by magnetic sorting with anti-CD138 antibodies and analyzed by SNParray and MLPA. Unsorted samples showed a lower incidence of abnormalities. By analyzing the data, we concluded that even a 30% malignant cell infiltration in the MM sample is not enough and the magnetic sorting is a mandatory for the enrichment of target cells from the sample. [ABSTRACT FROM AUTHOR]

Acute lymphoblastic leukemia represents a heterogeneous group of hematological malignancies, defined by clonal proliferation of lymphoid cells. Immunophenotyping by flow cytometry and molecular analysis for the detection of genetic anomalies are clinical standard procedures for diagnosis, sub-classification and post-therapeutic evaluation. Samples from 105 patients diagnosed with acute lymphoblastic leukemia were immunophenotyped at diagnosis and were investigated by molecular analysis in order to identify the occurrence of four fusion genes: MLL-AF4, TEL-AML-1, BCR-ABL-p190, E2A-PBX-1. There were no associations found between the immunophenotype and the presence of any fusion genes evaluated. Both methods in combination remain a prerequisite for an improved subclassification of hematological malignancies, therapeutic decision, and evaluation of treatment response. [ABSTRACT FROM AUTHOR]

In this study, new hybrid scaffolds based on 3D-printed titanium alloy with spongy bone-like architecture filled with porous hydroxyapatite (i.e. which also mimic compositional and micro geometric features of bone inorganic matrix) were investigated in vitro in terms of cell osteogenic differentiation, matrix maturation and mineralization. MTT-assay proved the high proliferation profile of the osteoblastic cells coincubated with the studied scaffold. RT-PCR evidenced enhanced expression level of several key osteogenic markers, such as Runt-related transcription factor 2, collagen, alkaline phosphatase, bone sialoprotein, osteocalcin. Extracellular matrix staining showed highly collagen synthesis. Alizarin-red staining confirmed nodule bone formation, i.e. matrix mineralization. The results put forward the osteogenic behaviour of the studied scaffolds and consequently their potential for using in bone restoration after further in vivo study. [ABSTRACT FROM AUTHOR]

The aim of this study was to monitor the influence of the fermentation conditions on the exopolysaccharides (EPS) biosynthesis. For this, different culture media compositions were tested on an isolated lactic acid bacteria (LAB) strain, identified by 16S rDNA sequence as being Weissella confusa . It was proved that this bacterial strain culture in MRS medium supplemented with 80 g/L sucrose and dissolved in UHT milk produced up to 25.2 g/L of freeze-dried EPS, in static conditions, after 48 h of fermentative process. Using FTIR and NMR analysis, it was demonstrated that the obtained EPS is a dextran. The thermal analysis revealed a dextran structure with high purity while GPC analysis depicted more fractions, which is normal for a biological obtained polymer. A concentration up to 3 mg/mL of dextran proved to have no cytotoxic effect on normal human dermal fibroblasts (NHDF). Moreover, at this concentration, dextran breaks up to 70% of the biofilms formed by the Candida albicans SC5314 strain, and has no antimicrobial activity against standard bacterial strains. Due to their characteristics, these EPS are suitable as hydrophilic matrix for controlled release of drugs in pharmaceutical industry. [ABSTRACT FROM AUTHOR]