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1.
Int J Mol Sci ; 24(2)2023 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-36674843

RESUMO

Skin wounds remain a significant problem for the healthcare system, affecting the clinical outcome, patients' quality of life, and financial costs. Reduced wound healing times would improve clinical, economic, and social aspects for both patients and the healthcare system. Skin wound healing has been studied for years, but effective therapy that leads to accelerated wound healing remains to be discovered. This study aimed to evaluate the potential of MELK silencing to accelerate wound healing. A vectorless, transient knockdown of the MELK gene using siRNA was performed in a murine skin wound model. The wound size, total collagen, type 3 collagen, vessel size, vessel number, cell proliferation, cell apoptosis, number of mast cells, and immune infiltration by CD45, CD11b, CD45, and CD8a cells were evaluated. We observed that treatment with MELK siRNA leads to significantly faster wound closing associated with increased collagen deposition.


Assuntos
Fibroblastos , Qualidade de Vida , Humanos , Animais , Camundongos , RNA Interferente Pequeno/genética , Cicatrização/genética , Colágeno/genética , Proliferação de Células/genética , Pele/lesões , Proteínas Serina-Treonina Quinases
2.
Int J Mol Sci ; 24(9)2023 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-37175795

RESUMO

Maternal embryonic leucine-zipper kinase (MELK) plays a significant role in cell cycle progression, mitosis, cell migration, cell renewal, gene expression, embryogenesis, proliferation, apoptosis, and spliceosome assembly. In addition, MELK is known to be overexpressed in multiple types of cancer and is associated with cancer proliferation. Tumorigenesis shares many similarities with wound healing, in which the rate of cell proliferation is a critical factor. Therefore, this study aimed to determine the involvement of MELK in the regulation of cell division in two cell types involved in this process, namely fibroblasts and keratinocytes. We examined how temporal overexpression of wild-type and kinase-dead MELK kinase variants affect the rate of proliferation, viability, cell cycle, and phosphorylation state of other kinases involved in these processes, such as ERK1/2, AKT1, MAPK9, p38, and p53. We explored if MELK could be used as a therapeutic stimulator of accelerated wound healing via increased proliferation. We observed that aberrant expression of MELK results in abnormal proliferation, altered cell cycle distribution, and decreased viability of the cells, which challenge the utility of MELK in accelerated wound healing. Our results indicate that, at least in healthy cells, any deviation from precisely controlled MELK expression is harmful to fibroblasts and keratinocytes.


Assuntos
Neoplasias , Proteínas Serina-Treonina Quinases , Humanos , Proteínas Serina-Treonina Quinases/metabolismo , Fosforilação , Proliferação de Células/genética , Queratinócitos/metabolismo , Linhagem Celular Tumoral
3.
Int J Mol Sci ; 25(1)2023 Dec 19.
Artigo em Inglês | MEDLINE | ID: mdl-38203201

RESUMO

Wounds represent a common occurrence in human life. Consequently, scientific investigations are underway to advance wound healing methodologies, with a notable focus on dressings imbued with biologically active compounds capable of orchestrating the wound microenvironment through meticulously regulated release mechanisms. Among these bioactive agents are cytokines, which, when administered to the wound milieu without appropriate protection, undergo rapid loss of their functional attributes. Within the context of this research, we present a method for fabricating dressings enriched with G-CSF (granulocyte colony-stimulating factor) or GM-CSF (granulocyte-macrophage colony-stimulating factor), showcasing both biological activity and protracted release dynamics. Based on Ligasano, a commercial polyurethane foam dressing, and chitosan crosslinked with TPP (sodium tripolyphosphate), these dressings are noncytotoxic and enable cytokine incorporation. The recovery of cytokines from dressings varied based on the dressing preparation and storage techniques (without modification, drying, freeze-drying followed by storage at 4 °C or freeze-drying followed by storage at 24 °C) and cytokine type. Generally, drying reduced cytokine levels and their bioactivity, especially with G-CSF. The recovery of G-CSF from unmodified dressings was lower compared to GM-CSF (60% vs. 80%). In summary, our freeze-drying approach enables the storage of G-CSF or GM-CSF enriched dressings at 24 °C with minimal cytokine loss, preserving their biological activity and thus enhancing future clinical availability.


Assuntos
Quitosana , Surdez , Humanos , Citocinas , Fator Estimulador de Colônias de Granulócitos e Macrófagos/farmacologia , Fator Estimulador de Colônias de Granulócitos , Bandagens
4.
Int J Mol Sci ; 24(6)2023 Mar 11.
Artigo em Inglês | MEDLINE | ID: mdl-36982453

RESUMO

Immortalized cell lines are widely used in vitro tools in oncology and hematology research. While these cell lines represent artificial systems and may accumulate genetic aberrations with each passage, they are still considered valuable models for pilot, preliminary, and screening studies. Despite their limitations, cell lines are cost-effective and provide repeatable and comparable results. Choosing the appropriate cell line for acute myeloid leukemia (AML) research is crucial for obtaining reliable and relevant results. Several factors should be considered when selecting a cell line for AML research, such as specific markers and genetic abnormalities associated with different subtypes of AML. It is also essential to evaluate the karyotype and mutational profile of the cell line, as these can influence the behavior and response to the treatment of the cells. In this review, we evaluate immortalized AML cell lines and discuss the issues surrounding them concerning the revised World Health Organization and the French-American-British classifications.


Assuntos
Leucemia Mieloide Aguda , Humanos , Leucemia Mieloide Aguda/metabolismo , Mutação , Cariotipagem , Cariótipo
5.
Int J Mol Sci ; 24(6)2023 Mar 16.
Artigo em Inglês | MEDLINE | ID: mdl-36982788

RESUMO

Natural compounds, such as resveratrol (Res), are currently used as adjuvants for anticancer therapies. To evaluate the effectiveness of Res for the treatment of ovarian cancer (OC), we screened the response of various OC cell lines to the combined treatment with cisplatin (CisPt) and Res. We identified A2780 cells as the most synergistically responding, thus optimal for further analysis. Because hypoxia is the hallmark of the solid tumor microenvironment, we compared the effects of Res alone and in combination with CisPt in hypoxia (pO2 = 1%) vs. normoxia (pO2 = 19%). Hypoxia caused an increase (43.2 vs. 5.0%) in apoptosis and necrosis (14.2 vs. 2.5%), reactive oxygen species production, pro-angiogenic HIF-1α (hypoxia-inducible factor-1α) and VEGF (vascular endothelial growth factor), cell migration, and downregulated the expression of ZO1 (zonula occludens-1) protein in comparison to normoxia. Res was not cytotoxic under hypoxia in contrast to normoxia. In normoxia, Res alone or CisPt+Res caused apoptosis via caspase-3 cleavage and BAX, while in hypoxia, it reduced the accumulation of A2780 cells in the G2/M phase. CisPt+Res increased levels of vimentin under normoxia and upregulated SNAI1 expression under hypoxia. Thus, various effects of Res or CisPt+Res on A2780 cells observed in normoxia are eliminated or diminished in hypoxia. These findings indicate the limitations in using Res as an adjuvant with CisPt therapy in OC.


Assuntos
Cisplatino , Neoplasias Ovarianas , Humanos , Feminino , Cisplatino/farmacologia , Cisplatino/uso terapêutico , Neoplasias Ovarianas/metabolismo , Resveratrol/farmacologia , Fator A de Crescimento do Endotélio Vascular/metabolismo , Linhagem Celular Tumoral , Hipóxia , Fatores de Crescimento do Endotélio Vascular/metabolismo , Hipóxia Celular , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Microambiente Tumoral
6.
Int J Mol Sci ; 23(16)2022 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-36012421

RESUMO

Animal research undoubtedly provides scientists with virtually unlimited data but inflicts pain and suffering on animals. Currently, legislators and scientists alike are promoting alternative in vitro approaches allowing for an accurate evaluation of processes occurring in the body without animal sacrifice. Historically, one of the most infamous animal tests is the Draize test, mainly performed on rabbits. Even though this test was considered the gold standard for around 50 years, the Draize test fails to mimic human response mainly due to human and rabbit eye physiological differences. Therefore, many alternative assays were developed to evaluate ocular toxicity and drug effectiveness accurately. Here we review recent achievements in tissue engineering of in vitro 2D, 2.5D, 3D, organoid and organ-on-chip ocular models, as well as in vivo and ex vivo models in terms of their advantages and limitations.


Assuntos
Alternativas aos Testes com Animais , Olho , Animais , Bioensaio , Humanos , Coelhos
7.
Molecules ; 27(17)2022 Aug 25.
Artigo em Inglês | MEDLINE | ID: mdl-36080193

RESUMO

Liquid soaps are the basic cosmetics used to clean the skin of the hands. Frequent hand washing prevents viral contamination but may damage the skin's hydro-lipid layer, leading to various types of irritation. Therefore, four liquid soap formulas were developed with three amphoteric surfactants: Cocamidopropyl Betaine (LS II), CocamidopropylHydroxysultaine (LS III), and newly synthesized Evening PrimroseaamidopropylSulfobetaine (LS IV). We evaluated the skin irritating potential (zein number, bovine albumin test) and cytotoxicity (AlamarBlue™, Cell viability, and Cell cycle assays) on HaCaT cell line. We observed lower values of the zein number and bovine albumin tests after adding soaps with surfactants (the highest differences in LS IV) compared to the base soap (LS I). However, LS I and LS II did not differ in cytotoxic assays. Therefore, adding LS III and LS IV seems potentially more dangerous to the cells. However, it should be noted that cells were continuously exposed to liquid soaps for more than 24 h, so its cytotoxic effects after dermal use in humans may be unnoticeable. Concluding, results suggest that the newly synthesized LS IV should improve the safety of liquid hand washing soaps.


Assuntos
Sabões , Zeína , Animais , Bovinos , Desinfecção das Mãos/métodos , Humanos , Soroalbumina Bovina , Sabões/farmacologia , Tensoativos/farmacologia
8.
Eur J Clin Microbiol Infect Dis ; 40(3): 541-547, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32986153

RESUMO

Clinical data suggest that during the current COVID-19 pandemic, children are less prone than adults to SARS-CoV-2 infection. Our purpose was to determine the frequency of SARS-CoV-2 in children vs. adults during the 2020 pandemic in Warsaw, Poland, and to investigate whether RSV and/or influenza A/B infections were associated with SARS-CoV-2 infections. We present results of RT-PCR tests for SARS-CoV-2 performed in Warsaw, Poland. Some of the pediatric subjects were also PCR-tested for RSV, and A and B influenza. We compared the test results from the four groups of symptomatic and asymptomatic subjects: 459 symptomatic pediatric patients (children 0-18 years old), 1774 symptomatic adults, 445 asymptomatic children, and 239 asymptomatic adults. 3.26% (15/459) of symptomatic pediatric patients were positive for SARS-CoV-2 in contrast to 5.58% (99/1774) of symptomatic adults (p = 0.0448). There were no SARS-CoV-2 positive cases in the group of asymptomatic children (0/445) and two positive cases in the group of asymptomatic adults (2/239), i.e., 0.83%. In the group of symptomatic pediatric patients, 17.14% (6/35) (p = 0.0002) were positive for RSV, 8.16% (4/49) were positive for influenza A, and 2.04% (1/49), thus 10.20% (5/49) (p = 0.0176) for influenza A/B. Children were less prone to SARS-CoV-2 infection than the adults during the COVID-19 pandemic in Warsaw. Higher percentage of symptomatic children was infected with RSV or influenza A/B than with SARS-CoV-2. This suggests a necessity for the testing for all these viruses for an early identification and isolation of SARS-CoV-2-positive patients for an ensuing 2020 autumn return of COVID-19.


Assuntos
COVID-19/diagnóstico , COVID-19/epidemiologia , SARS-CoV-2/isolamento & purificação , Adolescente , Adulto , Infecções Assintomáticas/epidemiologia , Criança , Pré-Escolar , Humanos , Lactente , Vírus da Influenza A/genética , Vírus da Influenza A/isolamento & purificação , Vírus da Influenza B/genética , Vírus da Influenza B/isolamento & purificação , Polônia/epidemiologia , Vírus Sinciciais Respiratórios/genética , Vírus Sinciciais Respiratórios/isolamento & purificação , Infecções Respiratórias/diagnóstico , Infecções Respiratórias/epidemiologia , Infecções Respiratórias/virologia , SARS-CoV-2/genética
9.
Int J Mol Sci ; 22(21)2021 Oct 27.
Artigo em Inglês | MEDLINE | ID: mdl-34769035

RESUMO

Background: The invention of non-ionizing emission devices revolutionized science, medicine, industry, and the military. Currently, different laser systems are commonly used, generating the potential threat of excessive radiation exposure, which can lead to adverse health effects. Skin is the organ most exposed to laser irradiation; therefore, this study aims to evaluate the effects of 445 nm, 520 nm, and 638 nm non-ionizing irradiation on keratinocytes and fibroblasts. Methods: Keratinocytes and fibroblasts were exposed to a different fluency of 445 nm, 520 nm, and 638 nm laser irradiation. In addition, viability, type of cell death, cell cycle distribution, and proliferation rates were investigated. Results: The 445 nm irradiation was cytotoxic to BJ-5ta (≥58.7 J/cm2) but not to Ker-CT cells. Exposure influenced the cell cycle distribution of Ker-CT (≥61.2 J/cm2) and BJ-5ta (≥27.6 J/cm2) cells, as well as the Bj-5ta proliferation rate (≥50.5 J/cm2). The 520 nm irradiation was cytotoxic to BJ-5ta (≥468.4 J/cm2) and Ker-CT (≥385.7 J/cm2) cells. Cell cycle distribution (≥27.6 J/cm2) of Ker-CT cells was also affected. The 638 nm irradiation was cytotoxic to BJ-5ta and Ker-CT cells (≥151.5 J/cm2). The proliferation rate and cell cycle distribution of BJ-5ta (≥192.9 J/cm2) and Ker-CT (13.8 and 41.3 J/cm2) cells were also affected. Conclusions: At high fluences, 455 nm, 520 nm, and 638 nm irradiation, representing blue, green, and red light spectra, are hazardous to keratinocytes and fibroblasts. However, laser irradiation may benefit the cells at low fluences by modulating the cell cycle and proliferation rate.


Assuntos
Fibroblastos/efeitos da radiação , Pele/efeitos da radiação , Ciclo Celular/efeitos da radiação , Morte Celular/efeitos da radiação , Proliferação de Células/efeitos da radiação , Sobrevivência Celular/efeitos da radiação , Células Cultivadas , Humanos , Lasers , Luz , Terapia com Luz de Baixa Intensidade/métodos
10.
Int J Mol Sci ; 22(5)2021 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-33670977

RESUMO

The invention of systems enabling the emission of waves of a certain length and intensity has revolutionized many areas of life, including medicine. Currently, the use of devices emitting laser light is not only an indispensable but also a necessary element of many diagnostic procedures. It also contributed to the development of new techniques for the treatment of diseases that are difficult to heal. The use of lasers in industry and medicine may be associated with a higher incidence of excessive radiation exposure, which can lead to injury to the body. The most exposed to laser irradiation is the skin tissue. The low dose laser irradiation is currently used for the treatment of various skin diseases. Therefore appropriate knowledge of the effects of lasers irradiation on the dermal cells' metabolism is necessary. Here we present current knowledge on the clinical and molecular effects of irradiation of different wavelengths of light (ultraviolet (UV), blue, green, red, and infrared (IR) on the dermal cells. .


Assuntos
Lasers , Luz , Pele/efeitos da radiação , Animais , Humanos , Raios Infravermelhos , Raios Ultravioleta
11.
Clin Immunol ; 217: 108510, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32544611

RESUMO

Children, because of having an immature immune system, are usually more prone than the adults to the microbial infections and have more severe symptoms, which is especially true for the newborns, and very young children. However, the review of clinical data from the current COVID-19 pandemic indicates otherwise. We discuss here what are the main features and components of children's immune system, the role of maternal transmission of immunity, and what are the possible explanations for the seemingly lower infection rate and severity of COVI-19 in children.


Assuntos
Betacoronavirus/imunologia , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/imunologia , Sistema Imunitário/virologia , Pandemias , Pneumonia Viral/epidemiologia , Pneumonia Viral/imunologia , Adulto , Fatores Etários , Betacoronavirus/patogenicidade , COVID-19 , Criança , Pré-Escolar , Infecções por Coronavirus/transmissão , Infecções por Coronavirus/virologia , Citocinas/imunologia , Resistência à Doença , Feminino , Humanos , Incidência , Lactente , Recém-Nascido , Leite Humano/imunologia , Placenta/imunologia , Placenta/virologia , Pneumonia Viral/transmissão , Pneumonia Viral/virologia , Gravidez , SARS-CoV-2
12.
Int J Mol Sci ; 21(13)2020 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-32629914

RESUMO

An important problem for researchers working in the field of dermatology is the preparation of the human skin equivalent (HSE). Here, we describe a simple and reliable protocol for preparing a skin model from the commercially available cell lines: keratinocytes, fibroblasts, and melanocytes. Importantly, in our 3D model, the keratinocytes are diverse that brings this model closer to the natural skin. For the production of HSE, we used available primary PCS-200-010, PCS-201-010, PCS-200-013, and immortalized CRL-4048 and CRL-4001 cell lines. We used genipin, which is necessary for collagen cross-linking and studied its cytotoxicity for keratinocytes and fibroblasts. The addition of 20 µM genipin reduced the shrinkage of the collagen in the constructs from 59% to 24% on day 12 of the culture of the construct. A higher concentration (80-200 µM) of genipin reduced shrinkage by 14% on average. Genipin in concentration 10 µM and below was not cytotoxic to the keratinocytes, and 150 µM and below to the fibroblasts. Hematoxylin and eosin staining showed that the morphology of HSEs was identical to that of native human skin. The immunohistochemical staining of the constructs showed the presence of vimentin-positive fibroblasts in the skin layer, while the melanocytes were in the epidermis and in the basal layer. We observed that the longer differentiation of constructs led to the higher secretion of GM-CSF, IL-10, IL-15, IL-1α, IL-6, IL-7, IL-8, and MCP-1. We also observed that the longer time of differentiation led to a more stable secretion of all analytes, which was reflected in the coefficient of variation. We described here a simple, reliable, and cost-effective production of the full-thickness human skin equivalents that can be used in the research and industry. With the global trend to decrease animal use for the research and testing, our HSE could be a useful testing tool and an alternative research model.


Assuntos
Técnicas de Cultura de Células/métodos , Pele/crescimento & desenvolvimento , Pele/metabolismo , Diferenciação Celular/fisiologia , Células Cultivadas , Colágeno/metabolismo , Derme/citologia , Células Epidérmicas/citologia , Epiderme/metabolismo , Matriz Extracelular/metabolismo , Fibroblastos/metabolismo , Humanos , Iridoides/farmacologia , Queratinócitos/metabolismo , Melanócitos/metabolismo , Modelos Biológicos , Pele Artificial
13.
Molecules ; 25(11)2020 May 27.
Artigo em Inglês | MEDLINE | ID: mdl-32471271

RESUMO

The aim of this study was to determine the cytotoxic properties, influence on enzyme activity involved in metabolic syndrome, and antimicrobial activity of synthetic peptides with GQLGEHGGAGMG, GEHGGAGMGGGQFQPV, EQGFLPGPEESGR, RLARAGLAQ, YGNPVGGVGH, and GNPVGGVGHGTTGT sequences. Peptides have no cytotoxic effect on cells. The highest inhibitory effect on angiotensin converting enzyme I was noted for peptide GT-14 (IC50 = 525.63 µg/mL). None of the tested peptides had an influence on α-glucosidase. The highest α-amylase and lipase inhibitory activity was noted for GG-12 (IC50 = 56.72 and 60.62 µg/mL, respectively). The highest lipoxidase inhibitory activity was determined for peptide ER-13 (IC50 = 84.35 µg/mL). Peptide RQ-9 was characterized by the highest COX inhibitory activity (0.31 and 4.77 µg/mL for COX-1 and COX-2, respectively). Only peptide RQ-9 inhibited S. enteritidis ATCC 4931 growth (42%-48%) in all tested concentrations (15.62-250 mg/mL).


Assuntos
Síndrome Metabólica/metabolismo , Peptídeos/química , Peptídeos/farmacologia , Anti-Infecciosos/química , Anti-Infecciosos/farmacologia , Lipase/metabolismo , Síndrome Metabólica/microbiologia , Salmonella enteritidis/efeitos dos fármacos , alfa-Amilases/metabolismo
14.
Cent Eur J Immunol ; 44(1): 23-32, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31114433

RESUMO

Vitamin B6 is necessary for many enzymatic pathways (glucose and lipid metabolism, DNA/RNA synthesis, or modulation of gene expression) and affects immune cell function and blood-forming processes. We hypothesised that supplementing a protein-deficient diet with vitamin B6 may reduce the negative impact of protein malnutrition. Here, we evaluated the effect of moderate, long-term exercise (ninety days) on selected blood parameters in rats fed a normal diet, a protein-deficient diet, or a protein-deficient diet supplemented with vitamin B6. Selected haematological, immunological, and biochemical parameters were examined. A protein-deficient diet lasting 90 days caused significant reduction in body mass, increased activity of aminotransferases (asparagine and alanine), an increased percentage of innate cells in the blood, and decreased haemoglobin concentration in the blood. Adding vitamin B6 significantly increased body and muscle mass, decreased liver parameters, and caused normalisation of haemoglobin concentration and the proportion of white blood cells in the blood. These results indicate that vitamin B6 supplementation significantly improves the health of protein-malnourished rats and paves the way for the development of novel anti-malnutrition therapies.

15.
Adv Exp Med Biol ; 1039: 35-44, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-28804811

RESUMO

Histamine intolerance (pseudoallergy) is a poorly investigated type of food hypersensitivity. The main enzyme responsible for histamine degradation in the extracellular matrix is diamine oxidase (DAO). Disturbances in the concentration or activity of DAO may lead to the development of clinical signs of allergy. The aim of the present work was to assess the DAO concentration, peripheral blood morphology, lymphocytes phenotyping (CD3+, CD4+, CD8+, CD19+, NK cells, NKT cells, and activated T-cells), and natural regulatory Treg (nTregs) cell population (CD4+, CD25+, CD127low, and FoxP3) in 34 pediatric patients with histamine-dependent syndromes. Patients were divided into two groups: classical allergy and pseudoallergy on the basis of IgE concentration. The investigation was based on the analysis of peripheral blood samples. A significantly lower serum DAO, both total and specific IgE, concentration was found in the pseudoallergy group compared with the allergy group. There were no significant differences in blood morphology or lymphocyte populations. A similar level of nTreg lymphocytes was also found in both groups, although it was lower than that present in healthy individuals. The findings suggest that the serum DAO is responsible for the symptoms of histamine intolerance. Moreover, a general decrease in nTreg cells in comparison with healthy individuals may lead to symptom aggravation.


Assuntos
Amina Oxidase (contendo Cobre)/sangue , Hipersensibilidade Alimentar/sangue , Hipersensibilidade/sangue , Adolescente , Criança , Pré-Escolar , Feminino , Hipersensibilidade Alimentar/imunologia , Humanos , Hipersensibilidade/imunologia , Imunoglobulina E/sangue , Lactente , Recém-Nascido , Ativação Linfocitária , Subpopulações de Linfócitos/imunologia , Linfócitos/imunologia , Masculino , Linfócitos T Reguladores/imunologia
16.
Radiat Environ Biophys ; 57(3): 251-264, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-29626227

RESUMO

Tritium is a potentially significant source of internal radiation exposure which, at high levels, can be carcinogenic. We evaluated whether single intraperitoneal injection of BALB/c and C57BL/6 mice with tritiated water (HTO) leading to exposure to low (0.01 or 0.1 Gy) and intermediate (1.0 Gy) cumulative whole-body doses of ß radiation is immunosuppressive, as judged by enhancement of artificial tumour metastases, functioning of NK lymphocytes and macrophages, circulating cytokine's levels, and numbers of bone marrow, spleen, and peripheral blood cells. We demonstrate that internal contamination of radiosensitive BALB/c and radioresistant C57BL/6 mice with HTO at all the absorbed doses tested did not affect the development of neoplastic colonies in the lungs caused by intravenous injection of syngeneic cancer cells. However, internal exposure of BALB/c and C57BL/6 mice to 0.1 and 0.01 Gy of ß radiation, respectively, up-regulated cytotoxic activity of and IFN-γ synthesis in NK lymphocytes and boosted macrophage secretion of nitric oxide. Internal contamination with HTO did not affect the serum levels of pro- (IL-1ß, IL-2, IL-6, TNF-α,) and anti-inflammatory (IL-1Ra, IL-4, IL-10) cytokines. In addition, exposure of mice of both strains to low and intermediate doses from the tritium-emitted ß-particles did not result in any significant changes in the numbers of bone marrow, spleen, and peripheral blood cells. Overall, our data indicate that internal tritium contamination of both radiosensitive and radioresistant mice leading to low and intermediate absorbed ß-radiation doses is not immunosuppressive but may enhance some but not all components of anticancer immunity.


Assuntos
Citocinas/metabolismo , Hematopoese/efeitos da radiação , Imunidade Inata/efeitos da radiação , Neoplasias Pulmonares/patologia , Tolerância a Radiação , Trítio/química , Água/farmacologia , Animais , Relação Dose-Resposta à Radiação , Neoplasias Pulmonares/sangue , Neoplasias Pulmonares/imunologia , Neoplasias Pulmonares/radioterapia , Masculino , Camundongos , Água/química
17.
Contemp Oncol (Pozn) ; 22(1A): 48-55, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29628794

RESUMO

Cancer stem-like cells (CSLCs) are defined as cancer cells with stem cell characteristics. Although CSLCs constitute no more than a few percent of the tumor mass, they play important roles in cancer chemo-resistance, metastasis and disease recurrence. Ovarian cancer (OC) is considered the most aggressive gynecological malignancy in which the role of CSLCs is of major significance, although it remains to be specified. The studies describing ovarian CSLC phenotype vary in the definition of the molecular pattern of expression of the main markers such as CD133, CD44, CD117, and CD24. Stem-like features of OC have been shown to correlate with the clinical course of the disease and permit diagnosis, prognosis and treatment outcome to be improved. Identification of CSLC markers could provide hallmarks which, related to the chemo-resistance of the disease, will facilitate treatment selection. This review describes recent advances in research on stem-like cell status in OC, mainly focusing on surface markers of CSLCs and their clinical relevance.

18.
BMC Cancer ; 17(1): 21, 2017 01 05.
Artigo em Inglês | MEDLINE | ID: mdl-28056882

RESUMO

BACKGROUND: CD105 was postulated as a renal cell carcinoma (RCC) stem cell marker, and CD133 as a putative RCC progenitor. Hypoxia, a natural microenvironment that prevails in tumors, was also incorporated into the study, especially in terms of the promotion of hypothetical stem-like cell properties. METHODS: Within this study, we verify the existence of CD105+ and CD133+ populations in selected papillary subtype RCC (pRCC) cell lines. Both populations were analyzed for correlation with stem-like cell properties, such as stemness gene expression, and sphere and colony formation. For the preliminary analysis, several RCC cell lines were chosen (786-O, SMKT-R2, Caki-2, 796-P, ACHN, RCC6) and the control was human kidney cancer stem cells (HKCSC) and renal cells of embryonic origin (ASE-5063). Four cell lines were chosen for further investigation: Caki-2 (one of the highest numbers of CD105+ cells; primary origin), ACHN (a low number of CD105+ cells; metastatic origin), HKCSC (putative positive control), and ASE-5063 (additional control). RESULTS: In 769-P and RCC6, we could not detect a CD105+ population. Hypoxia variously affects pRCC cell growth, and mainly diminishes the stem-like properties of cells. Furthermore, we could not observe the correlation of CD105 and/or CD133 expression with the enhancement of stem-like properties. CONCLUSIONS: Based on this analysis, CD105/CD133 cannot be validated as cancer stem cell markers of pRCC cell lines.


Assuntos
Carcinoma Papilar/patologia , Carcinoma de Células Renais/patologia , Endoglina/análise , Neoplasias Renais/patologia , Células-Tronco Neoplásicas/patologia , Linhagem Celular Tumoral , Separação Celular , Perfilação da Expressão Gênica , Humanos , Imuno-Histoquímica , Reação em Cadeia da Polimerase em Tempo Real , Transcriptoma
19.
Med Sci Monit ; 23: 4865-4873, 2017 Oct 11.
Artigo em Inglês | MEDLINE | ID: mdl-29018182

RESUMO

BACKGROUND Understanding the mechanisms conditioning development of chronic kidney disease (CKD) is still a challenge. The aim of this study was to evaluate the activity of the intrarenal nitric oxide (NO) pathway in the context of sensitivity or resistance of different animal strains to the development and degree of renal failure. MATERIAL AND METHODS Two rat strains were used: Wistar (WR) and Sprague-Dawley rats (SDR) in a model of CKD - 5/6 nephrectomy. We assessed parameters of renal failure and expression of nitric oxide synthase (NOS) isoforms in renal cortex and medulla. RESULTS We did not observe renal failure in WR, and CKD developed in SDR with increase of creatinine and urea concentration as well as decrease of diuresis and glomerular filtration. In the renal cortex, baseline expression of NOS2 was higher in WR than in SDR. 5/6 nephrectomy resulted in reduction of NOS2 in both strains and NOS3 in WR. In the renal medulla, baseline NOS2 expression was higher in SDR, and nephrectomy resulted in its decrease only in SDR. Although baseline NOS3 expression was higher in SDR, the NOS3 expression after nephrectomy was higher in WR rats. CONCLUSIONS In model of CKD - 5/6 nephrectomy, SDR proved to be sensitive and WR resistant to development of CKD. The intrarenal activity of the nitric oxide pathway was the factor that differentiated both strains. This mechanism may be responsible for insensitivity of WR to development of renal failure in this model of CKD.


Assuntos
Óxido Nítrico Sintase/fisiologia , Insuficiência Renal Crônica/metabolismo , Insuficiência Renal Crônica/fisiopatologia , Animais , Creatinina/metabolismo , Modelos Animais de Doenças , Progressão da Doença , Rim/metabolismo , Falência Renal Crônica/metabolismo , Masculino , Modelos Teóricos , Nefrectomia/métodos , Óxido Nítrico/metabolismo , Óxido Nítrico/fisiologia , Óxido Nítrico Sintase/metabolismo , Óxido Nítrico Sintase Tipo I/metabolismo , Óxido Nítrico Sintase Tipo III/metabolismo , Isoformas de Proteínas , Ratos , Ratos Sprague-Dawley/fisiologia , Ratos Wistar/fisiologia , Insuficiência Renal/metabolismo
20.
Adv Exp Med Biol ; 1020: 81-89, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28512681

RESUMO

The influence of vitamin D on allergic diseases, including atopic dermatitis, is linked to the presence of vitamin D nuclear receptors in immune cells. The present study seeks to determine the possible relationship between serum vitamin D content and immune indices in children with atopic dermatitis. The study was conducted in 19 children with atopic dermatitis. The control consisted of 17 age-matched healthy children. A single significant finding was a distinctly lower number of serum regulatory T cells in atopic dermatitis compared with controls (p < 0.00001). There were no appreciable differences between the two groups concerning the immunological indices such as the phenotypes: CD3, CD4, CD8, CD4/CD8, CD19, CD16/56, natural killer T cells, and anti-CD3 human leukocyte antigen - antigen D related cell surface receptor (HLA-DR3), or the percentage of lymphocytes, eosinophils, and the IgE level. We also revealed an inverse association between the serum vitamin D and the percentage of CD8+ cells (p < 0.05; r = 0.62) in atopic dermatitis. In conclusion, the results point to a regulatory role of T cells in the pathogenesis of atopic dermatitis, but fail to substantiate the influence of vitamin D on the course of the disease.


Assuntos
Dermatite Atópica/sangue , Dermatite Atópica/imunologia , Vitamina D/sangue , Estudos de Casos e Controles , Criança , Humanos , Imunoglobulina E/sangue , Linfócitos T/imunologia
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