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Background: Radiation therapy has undergone significant technical development in the past decade. However, the complex therapy of intermediate-risk patients with organ-confined prostate carcinoma still poses many questions. Our retrospective study investigated the impact of selected components of the treatment process including radiotherapy, hormone deprivation, risk classification, and patients’ response to therapy. Methods: The impact of delivered dose, planning accuracy, duration of hormone deprivation, risk classification, and the time to reach prostate-specific antigen (PSA) nadir state were analyzed among ninety-nine individuals afflicted with organ-confined disease. Progression was defined as a radiological or biochemical relapse within five years from radiotherapy treatment. Results: We found that 58.3% of the progressive population consisted of intermediate-risk patients. The progression rate in the intermediate group was higher (21.9%) than in the high-risk population (12.1%). Dividing the intermediate group, according to the International Society of Urological Pathology (ISUP) recommendations, resulted in the non-favorable subgroup having the highest rate of progression (33.3%) and depicting the lowest percentage of progression-free survival (66.7%). Conclusion: Extended pelvic irradiation on the regional lymph nodes may be necessary for the ISUP Grade 3 subgroup, similarly to the high-risk treatment. Therapy optimization regarding the intermediate-risk population based on the ISUP subgrouping suggestions is highly recommended in the treatment of organ-confined prostate cancer.
Viktória Temesfői; Róbert Herczeg; Zoltán Lőcsei; Klára Sebestyén; Zsolt Sebestyén; László Mangel; Miklós Damásdi. Should We Reconsider the Necessity of a Refinement of Prostate Cancer Risk Classification and Radiotherapy Treatment Strategy? Experiences from a Retrospective Analysis of Data from a Single Institution. Journal of Clinical Medicine 2020, 10, 110 .
AMA StyleViktória Temesfői, Róbert Herczeg, Zoltán Lőcsei, Klára Sebestyén, Zsolt Sebestyén, László Mangel, Miklós Damásdi. Should We Reconsider the Necessity of a Refinement of Prostate Cancer Risk Classification and Radiotherapy Treatment Strategy? Experiences from a Retrospective Analysis of Data from a Single Institution. Journal of Clinical Medicine. 2020; 10 (1):110.
Chicago/Turabian StyleViktória Temesfői; Róbert Herczeg; Zoltán Lőcsei; Klára Sebestyén; Zsolt Sebestyén; László Mangel; Miklós Damásdi. 2020. "Should We Reconsider the Necessity of a Refinement of Prostate Cancer Risk Classification and Radiotherapy Treatment Strategy? Experiences from a Retrospective Analysis of Data from a Single Institution." Journal of Clinical Medicine 10, no. 1: 110.
Medicinal plants are widely used in folk medicine but quite often their composition and biological effects are hardly known. Our study aimed to analyze the composition, cytotoxicity, antimicrobial, antioxidant activity and cellular migration effects of Anthyllis vulneraria, Fuchsia magellanica, Fuchsia triphylla and Lysimachia nummularia used in the Romanian ethnomedicine for wounds. Liquid chromatography with mass spectrometry (LC-MS/MS) was used to analyze 50% (v/v) ethanolic and aqueous extracts of the plants’ leaves. Antimicrobial activities were estimated with a standard microdilution method. The antioxidant properties were evaluated by validated chemical cell-free and biological cell-based assays. Cytotoxic effects were performed on mouse fibroblasts and human keratinocytes with a plate reader-based method assessing intracellular adenosine triphosphate (ATP), nucleic acid and protein contents and also by a flow cytometer-based assay detecting apoptotic–necrotic cell populations. Cell migration to cover cell-free areas was visualized by time-lapse phase-contrast microscopy using standard culture inserts. Fuchsia species showed the strongest cytotoxicity and the highest antioxidant and antimicrobial activity. However, their ethanolic extracts facilitated cell migration, most probably due to their various phenolic acid, flavonoid and anthocyanin derivatives. Our data might serve as a basis for further animal experiments to explore the complex action of Fuchsia species in wound healing assays.
Rita Csepregi; Viktória Temesfői; Sourav Das; Ágnes Alberti; Csenge Anna Tóth; Róbert Herczeg; Nóra Papp; Tamás Kőszegi. Cytotoxic, Antimicrobial, Antioxidant Properties and Effects on Cell Migration of Phenolic Compounds of Selected Transylvanian Medicinal Plants. Antioxidants 2020, 9, 166 .
AMA StyleRita Csepregi, Viktória Temesfői, Sourav Das, Ágnes Alberti, Csenge Anna Tóth, Róbert Herczeg, Nóra Papp, Tamás Kőszegi. Cytotoxic, Antimicrobial, Antioxidant Properties and Effects on Cell Migration of Phenolic Compounds of Selected Transylvanian Medicinal Plants. Antioxidants. 2020; 9 (2):166.
Chicago/Turabian StyleRita Csepregi; Viktória Temesfői; Sourav Das; Ágnes Alberti; Csenge Anna Tóth; Róbert Herczeg; Nóra Papp; Tamás Kőszegi. 2020. "Cytotoxic, Antimicrobial, Antioxidant Properties and Effects on Cell Migration of Phenolic Compounds of Selected Transylvanian Medicinal Plants." Antioxidants 9, no. 2: 166.
A fluorescence-based enzymatic microplate intracellular glucose assay was designed and fully validated. The method was tested in a hepatocellular cancer cell line (HepG2). Our novel one-step extraction reagent gave stable cell lysates for glucose, adenosine triphosphate (ATP), and total protein determination from the same sample. Limit of detection for glucose was 0.13 µM (26 pmol/well), which is superior to commercially available glucose assays. Both intra- and interday assay imprecision in HepG2 cultures were less than 12% coefficient of variance (CV). In cell lysates spiked with glucose, recovery at two levels varied between 83.70% and 91.81%, and both linearity and stability were acceptable. HepG2 cells treated with agents affecting glucose uptake/metabolism (phloretin, quercetin, quercetin-3′-sulfate, NaF, 3-bromopyruvate, NaN3, oligomycin A, ochratoxin A, cytochalasin B, and anti-GLUT1 antibody) showed dose-dependent changes in glucose and ATP levels without total protein (cell) loss. Finally, we performed flow cytometric glucose uptake measurement in the treated cells using 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxyglucose fluorescent glucose analog. Glucose uptake did not always mirror the intracellular glucose levels, which most likely reflects the differences between the two methodologies. However, interpreting data obtained by both methods and taking ATP/protein levels at the same time, one can get information on the mode of action of the compounds.
Rita Csepregi; Viktória Temesfői; Nikolett Sali; Miklós Poór; Paul W. Needs; Paul A. Kroon; Tamás Kőszegi. A One-Step Extraction and Luminescence Assay for Quantifying Glucose and ATP Levels in Cultured HepG2 Cells. International Journal of Molecular Sciences 2018, 19, 2670 .
AMA StyleRita Csepregi, Viktória Temesfői, Nikolett Sali, Miklós Poór, Paul W. Needs, Paul A. Kroon, Tamás Kőszegi. A One-Step Extraction and Luminescence Assay for Quantifying Glucose and ATP Levels in Cultured HepG2 Cells. International Journal of Molecular Sciences. 2018; 19 (9):2670.
Chicago/Turabian StyleRita Csepregi; Viktória Temesfői; Nikolett Sali; Miklós Poór; Paul W. Needs; Paul A. Kroon; Tamás Kőszegi. 2018. "A One-Step Extraction and Luminescence Assay for Quantifying Glucose and ATP Levels in Cultured HepG2 Cells." International Journal of Molecular Sciences 19, no. 9: 2670.
Green fluorescent protein (GFP) is considered to be suitable for cell viability testing. In our study, GFP transfected A549 lung carcinoma cell line was treated with sodium fluoride (NaF), cycloheximide (CHX) and ochratoxin A (OTA). GFP fluorescence, intracellular ATP, nucleic acid and protein contents were quantified by a luminescence microplate assay developed in our laboratory. Flow cytometry was used to confirm the findings and to assess the intensity of GFP during different types of cell death. A 24 h NaF and CHX exposure caused a dramatic decrease in ATP contents (p < 0.05) compared with those of the controls. GFP fluorescence of the cells was in close correlation with total protein; however, GFP/ATP increased at NaF and decreased at CHX treatments (p < 0.05). ATP/protein and ATP/propidium iodide (PI) were largely decreased at NaF exposure in a dose-dependent manner (p < 0.05), while CHX and OTA showed markedly fewer effects. Both treatments caused apoptosis/necrosis at different rates. NaF induced mainly late apoptosis while OTA, mainly apoptosis. CHX effects varied by the incubation time with 100-fold elevation in late apoptotic cells at 24 h treatment. GFP intensity did not show a significant difference between live and apoptotic populations. Our results suggest when using GFP, a multiparametric assay is necessary for more precise interpretation of cell viability.
Rita Csepregi; Viktória Temesfői; Miklós Poór; Zsuzsanna Faust; Tamás Kõszegi. Green Fluorescent Protein-Based Viability Assay in a Multiparametric Configuration. Molecules 2018, 23, 1575 .
AMA StyleRita Csepregi, Viktória Temesfői, Miklós Poór, Zsuzsanna Faust, Tamás Kõszegi. Green Fluorescent Protein-Based Viability Assay in a Multiparametric Configuration. Molecules. 2018; 23 (7):1575.
Chicago/Turabian StyleRita Csepregi; Viktória Temesfői; Miklós Poór; Zsuzsanna Faust; Tamás Kõszegi. 2018. "Green Fluorescent Protein-Based Viability Assay in a Multiparametric Configuration." Molecules 23, no. 7: 1575.
To clarify controversies in the literature of the field, we have purified and characterized B16F1 melanoma cell derived exosomes (mcd-exosomes) then we attempted to dissect their immunological activities. We tested how mcd-exosomes influence CD4+ T cell proliferation induced by bone marrow derived dendritic cells; we quantified NF-κB activation in mature macrophages stimulated with mcd-exosomes, and we compared the cytokine profile of LPS-stimulated, IL-4 induced, and mcd-exosome treated macrophages. We observed that mcd-exosomes helped the maturation of dendritic cells, enhancing T cell proliferation induced by the treated dendritic cells. The exosomes also activated macrophages, as measured by NF-κB activation. The cytokine and chemokine profile of macrophages treated with tumor cell derived exosomes showed marked differences from those induced by either LPS or IL-4, and it suggested that exosomes may play a role in the tumor progression and metastasis formation through supporting tumor immune escape mechanisms.
Annamaria Marton; Csaba Vizler; Erzsebet Kusz; Viktoria Temesfoi; Zsuzsa Szathmary; Krisztina Nagy; Zsolt Szegletes; Gyorgy Varo; Laszlo Siklos; Robert L. Katona; Vilmos Tubak; O.M. Zack Howard; Erno Duda; Janos Minarovits; Katalin Nagy; Krisztina Buzas. Melanoma cell-derived exosomes alter macrophage and dendritic cell functions in vitro. Immunology Letters 2012, 148, 34 -38.
AMA StyleAnnamaria Marton, Csaba Vizler, Erzsebet Kusz, Viktoria Temesfoi, Zsuzsa Szathmary, Krisztina Nagy, Zsolt Szegletes, Gyorgy Varo, Laszlo Siklos, Robert L. Katona, Vilmos Tubak, O.M. Zack Howard, Erno Duda, Janos Minarovits, Katalin Nagy, Krisztina Buzas. Melanoma cell-derived exosomes alter macrophage and dendritic cell functions in vitro. Immunology Letters. 2012; 148 (1):34-38.
Chicago/Turabian StyleAnnamaria Marton; Csaba Vizler; Erzsebet Kusz; Viktoria Temesfoi; Zsuzsa Szathmary; Krisztina Nagy; Zsolt Szegletes; Gyorgy Varo; Laszlo Siklos; Robert L. Katona; Vilmos Tubak; O.M. Zack Howard; Erno Duda; Janos Minarovits; Katalin Nagy; Krisztina Buzas. 2012. "Melanoma cell-derived exosomes alter macrophage and dendritic cell functions in vitro." Immunology Letters 148, no. 1: 34-38.