The most commonly used flow cytometric (FCM) analysis of cellular DNA content relies on ethanol fixation followed by RNA digestion and propidium iodide (PI) intercalation into double-stranded DNA. This is a laborious and time-consuming procedure that is subject to systematic errors due to centrifugation and washing steps associated with sample preparation. It can adversely affect the reliability of the results. Here, we present a modified concept of DNA quantification in adherent cell lines by FCM that involves neither ethanol fixation nor any washing and cell transferring steps. Our high throughput assay of adherent cell lines reduces sample-processing time, requires minimal workload, provides a possibility for automation, and, if needed, also allows a significant reduction in the size of individual samples. Working with a well-proven commercial tool-The BD CycletestTM Plus DNA Reagent Kit-primarily designed for cell cycle analysis and aneuploidy determination in experimental and clinical samples, we suggest a novel, very efficient, and robust approach for DNA research in adherent cell cultures.
- MeSH
- aneuploidie MeSH
- automatizace MeSH
- buněčná adheze MeSH
- buněčný cyklus genetika MeSH
- DNA * analýza MeSH
- lidé MeSH
- průtoková cytometrie * metody MeSH
- reprodukovatelnost výsledků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
The role of postmastectomy radiotherapy and regional nodal irradiation after radical mastectomy is defined in high-risk patients with locally advanced tumors, positive margins, and unfavorable biology. The benefit of postmastectomy radiotherapy in intermediate-risk patients (T3N0 tumors) remains a matter of controversy. It has been demonstrated that radiotherapy after breast-conserving surgery lowers the locoregional recurrence rate compared with surgery alone and improves the overall survival rate. In patients with four or more positive lymph nodes or extracapsular extension, regional lymph node irradiation is indicated regardless of the surgery type (breast-conserving surgery or mastectomy). Despite the consensus that patients with more than three positive lymph nodes should be treated with radiotherapy, there is controversy regarding the recommendations for patients with one to three involved lymph nodes. In patients with N0 disease with negative findings on axillary surgery, there is a trend to administer regional lymph node irradiation in patients with a high risk of recurrence. In patients treated with neoadjuvant systemic therapy and mastectomy, adjuvant radiotherapy should be administered in cases of clinical stage III and/or ≥ypN1. In patients treated with neoadjuvant systemic therapy and breast-conserving surgery, postoperative radiotherapy is indicated irrespective of pathological response.
- MeSH
- adjuvantní radioterapie MeSH
- lidé MeSH
- lokální recidiva nádoru patologie MeSH
- mastektomie MeSH
- nádory prsu * farmakoterapie MeSH
- segmentální mastektomie MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Modern radiotherapy techniques are designed to permit reduced irradiation of healthy tissue, resulting in a diminished risk of adverse effects and shortened recovery times. Several randomized studies have demonstrated the benefits of increased dosage to the tumor bed area in combination with whole breast irradiation (WBI). Conventional WBI treatment following breast-conserving procedures, which required 5-7 weeks of daily treatments, has been reduced to 3-4 weeks when using hyperfractionated regimens. The dosage administration improves local control, albeit with poorer cosmesis. The method of accelerated partial breast irradiation (APBI) shortens the treatment period whilst reducing the irradiated volume. APBI can be delivered using intraoperative radiation, brachytherapy, or external beam radiotherapy. Currently available data support the use of external beam partial breast irradiation in selected patients. Modern radiotherapy techniques make it possible to achieve favorable cosmesis in most patients undergoing immediate breast reconstruction surgery, and studies confirm that current methods of external beam radiation allow an acceptable coverage of target volumes both in the reconstructed breast and in the regional lymphatic nodes.
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
This study establishes a new experimental approach for retrospective biodosimetric assessment by apoptosis detection ex vivo. For this purpose, we used mononuclear blood leukocytes isolated from the peripheral blood of irradiated Wistar rats and cultured them ex vivo for posterior analysis. Using flow cytometry, we distinguished apoptotic lymphocyte subsets individual biodosimetric potential at different time periods after exposure: B-lymphocytes 6-8 h (0-7 Gy), natural killer cells 24 h (0-7 Gy) and T-lymphocytes 24 h (0-1 Gy). This novel experimental design innovates through the need of a single blood sample from irradiated individuals for a complete biodosimetric assessment.
The dramatic rise in diagnostic procedures, radioisotope-based scans and intervention procedures has created a very valid concern regarding the long-term biological consequences from exposure to low doses of ionizing radiation. Despite its unambiguous medical benefits, additional knowledge on the health outcome of its use is essential. This review summarizes the available information regarding the biological consequences of low-dose radiation (LDR) exposure in humans (e.g. cytogenetic changes, cancer risk and radiation-induced cataracts. However, LDR studies remain relatively new and thus an encompassing view of its biological effects and relevant mechanisms in the human body is still needed.
- MeSH
- dávka záření MeSH
- ionizující záření * MeSH
- lidé MeSH
- počítačová rentgenová tomografie metody MeSH
- radiační poranění * etiologie prevence a kontrola MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
The JC-1 dye is widely used in apoptosis studies to monitor mitochondrial health. The probe was tested in vitro on two established cell lines and peripheral porcine blood lymphocytes after gamma irradiation (IR) to assess its potential in biodosimetric evaluation. In brief, we stained irradiated and non-irradiated cells with the JC-1 dye to determine the existing changes in mitochondrial membrane potential and monitor cell health through flow cytometry. The stage of injury in these cells was evaluated through an irradiated versus non-irradiated ratio (IVNIR), comparing the relative proportion of polarised cells containing red JC-1 aggregates. We observed a decreasing IVNIR as the radiation dose increased (i.e. 0.5; 1; 2; 4; 6; 8 and 10 Gy), performing the analysis at 4, 8 and 24 h after IR in all the tested cells. The results from the JC1-dye test showed that CD4 T lymphocytes were more sensitive to irradiation than other subpopulations.
Hyaluronic acid (HA) has a special position among glycosaminoglycans. As a major component of the extracellular matrix (ECM). This simple, unbranched polysaccharide is involved in the regulation of various biological cell processes, whether under physiological conditions or in cases of cell damage. This review summarizes the history of this molecule's study, its distinctive metabolic pathway in the body, its unique properties, and current information regarding its interaction partners. Our main goal, however, is to intensively investigate whether this relatively simple polymer may find applications in protecting against ionizing radiation (IR) or for therapy in cases of radiation-induced damage. After exposure to IR, acute and belated damage develops in each tissue depending upon the dose received and the cellular composition of a given organ. A common feature of all organ damage is a distinct change in composition and structure of the ECM. In particular, the important role of HA was shown in lung tissue and the variability of this flexible molecule in the complex mechanism of radiation-induced lung injuries. Moreover, HA is also involved in intermediating cell behavior during morphogenesis and in tissue repair during inflammation, injury, and would healing. The possibility of using the HA polymer to affect or treat radiation tissue damage may point to the missing gaps in the responsible mechanisms in the onset of this disease. Therefore, in this article, we will also focus on obtaining answers from current knowledge and the results of studies as to whether hyaluronic acid can also find application in radiation science.
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
PURPOSE: This paper compares individual radiation therapy techniques used for prostate cancer and their benefits in clinical practice. METHODS: We retrospectively analyzed 921 patients with localized prostate tumors treated between 1997 and 2012. We divided the patients into four groups according to the selected treatment technique (conformal radiation therapy [3DCRT], intensity-modulated radiation therapy [IMRT], image-guided radiation therapy [IGRT], and volumetric-modulated arc therapy [VMAT]) and evaluated the incidence of acute and chronic gastrointestinal (GI) and genitourinary (GU) toxicity. RESULTS: The incidence of grade 2 or greater acute GU and GI toxicity was significantly higher among techniques other than IGRT (p˂0.001). We found the same results in the case of grade 3 or greater acute GU toxicity (p˂0.001). Grade 3 or higher acute GI toxicity occurred only in one patient treated by 3DCRT. Cumulative late GI toxicity of grade 2 or higher and grade 3 or higher was recorded over 3 years significantly more frequently among non-IGRT techniques as compared to IGRT (p˂0.001). As regards GU toxicity, we found significantly higher incidence only for grade 2 or higher (p˂0.001), not for grade 3 or higher. No occurrence of grade 4 toxicity was recorded. The greatest incidence of patients without acute and chronic GI/GU toxicity was recorded in connection with VMAT. CONCLUSION: IGRT demonstrated a pronounced reduction in acute and chronic GU and GI toxicity as compared to non-IGRT techniques in the treatment of localized prostate cancer.
- MeSH
- akutní nemoc MeSH
- chronická nemoc MeSH
- gastrointestinální trakt patofyziologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- nádory prostaty komplikace MeSH
- radioterapie řízená obrazem metody MeSH
- retrospektivní studie MeSH
- senioři MeSH
- urogenitální systém patofyziologie MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- Publikační typ
- časopisecké články MeSH
Purpose: Among other reasons, the deteriorating global security situation and dangers associated with nuclear weapons have increased the need for deeper knowledge of the basic mechanisms involving the human immune system and ionizing radiation (IR). We conducted a review as to the effects of IR on thymic tissue, and particularly on the development of thymocytes and the T lymphocytes population in peripheral blood. Existing knowledge on this topic is based in part on national registers that store records concerning irradiated people. The majority of studies in this area, however, are based on experimental animal models. The main open question in this subject area regards the delayed effects of IR on thymus tissue, development of thymocytes, and subsequent impact on the immune system. Findings acquired to date on effects of IR are contributing to emerging fields such as immunotherapy, the objective of which is to support or activate natural immunity response. Methods: Recent research articles were reviewed regarding the influence of IR on thymus tissue and thymocytes development. Results: Differentiation and proliferation of thymocytes constitute a complex and sensitive process that is partially altered after irradiation, as are, too, the mechanisms for movement of early (derived from bone marrow) and derived (thymus derivatives) precursors. Disruption of these processes may lead to alteration of immune system function. Conclusions: Low doses (<200 mGy) may lead to changes in or disruption of functions of the thymus, thymocytes, and mechanisms of the immune system. The extent of IR’s influence is dependent not only on the individual’s radiosensitivity but also on his or her sex and age. With increasing absorbed IR dose, the risk of damage to thymus tissue and thymocytes in the organism rises and the extent of damage increases.
- MeSH
- dávka záření MeSH
- hematopoéza účinky záření MeSH
- imunitní systém účinky záření MeSH
- ionizující záření * MeSH
- lidé MeSH
- lymfocyty účinky záření MeSH
- radiační expozice * škodlivé účinky MeSH
- radiační účinky MeSH
- reiradiace MeSH
- thymocyty účinky záření MeSH
- thymus účinky záření MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy MeSH
