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1.
Int J Mol Sci ; 24(5)2023 Mar 03.
Artigo em Inglês | MEDLINE | ID: mdl-36902343

RESUMO

Transcriptional analyses such as microarray data have contributed to the progress in the diagnostics and therapy of colorectal cancer (CRC). The need for such research is still present because of the disease being common in both men and women with a high second position in cancer rankings. Little is known about the relations between the histaminergic system and inflammation in the large intestine and CRC. Therefore, the aim of this study was to evaluate the expression of genes related to the histaminergic system and inflammation in the CRC tissues at three cancer development designs: all tested CRC samples, low (LCS) and high (HCS) clinical stage, and four clinical stages (CSI-CSIV), to the control. The research was carried out at the transcriptomic level, analysing hundreds of mRNAs from microarrays, as well as carrying out RT-PCR analysis of histaminergic receptors. The following histaminergic mRNAs: GNA15, MAOA, WASF2A, and inflammation-related: AEBP1, CXCL1, CXCL2, CXCL3, CXCL8, SPHK1, TNFAIP6, were distinguished. Among all analysed transcripts, AEBP1 can be considered the most promising diagnostic marker in the early stage of CRC. The results showed 59 correlations between differentiating genes of the histaminergic system and inflammation in the control, control and CRC, and CRC. The tests confirmed the presence of all histamine receptor transcripts in both the control and colorectal adenocarcinoma. Significant differences in expression were stated for HRH2 and HRH3 in the advanced stages of CRC adenocarcinoma. The relations between the histaminergic system and inflammation-linked genes in both the control and the CRC have been observed.


Assuntos
Adenocarcinoma , Neoplasias Colorretais , Masculino , Humanos , Feminino , Intestino Grosso/metabolismo , Neoplasias Colorretais/patologia , Inflamação , Adenocarcinoma/patologia , Perfilação da Expressão Gênica , Carboxipeptidases , Proteínas Repressoras/genética
2.
J Clin Med ; 9(12)2020 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-33322704

RESUMO

BACKGROUND: Autophagy plays a dual role of tumor suppression and tumor promotion in colorectal cancer. The study aimed to find those microRNAs (miRNAs) important in BECN1, LAMP2, and PINK1 regulation and to determine the possible role of the epigenetic changes in examined colorectal cancer using an in silico approach. METHODS: A total of 44 pairs of surgically removed tumors at clinical stages I‒IV and healthy samples (marginal tissues) from patients' guts were analyzed. Analysis of the obtained results was conducted using the PL-Grid Infrastructure and Statistica 12.0 program. The miRNAs and CpG islands were estimated using the microrna.org database and MethPrimer program. RESULTS: The autophagy-related genes were shown to be able to be regulated by miRNAs (BECN1-49 mRNA, LAMP2-62 mRNA, PINK1-6 mRNA). It was observed that promotion regions containing at least one CpG region were present in the sequence of each gene. CONCLUSIONS: The in silico analysis performed allowed us to determine the possible role of epigenetic mechanisms of regulation gene expression, which may be an interesting therapeutic target in the treatment of colorectal cancer.

3.
J Hum Kinet ; 73: 73-82, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32774539

RESUMO

The objective of the study to determine the effects of graded exercise on the cytokines and insulin-like growth factor-1 (IGF-1), growth hormone (hGH), testosterone (T), and cortisol (C) concentrations in the peripheral blood of female soccer players, and to evaluate if increased inflammatory biomarkers were related to these hormones and performance variables. Sixteen female soccer players (N = 16, age 19.3 ± 2.3 years) participated in this study. Blood samples were collected at three time points: pre-exercise, post-exercise, and in the 15th minute of recovery, to evaluate morphological and biochemical variables. The relative expression of IL-6 (interleukin 6) and serum concentrations of the cytokines were increased in the recovery period compared to pre-exercise levels (p = 0.03 and p=0.005, respectively). There was a significant effect of exercise on serum hGH level (p " 0.001), T/C ratio (p = 0.001), and C level (p=0.02). Positive correlations were found between: post-exercise IL-1ß (interleukin 1 beta) and IL-6 (R = 0.84, p = 0.000), and the IL-6 and TNF-α (tumor necrosis factor alpha) gene expression during recovery (R = 0.65, p = 0.009), and serum IL-1ß post-exercise and maximal power (R = 0.68; p = 0.004). Exercise-induced serum C levels positively correlated with IGF-1 levels (R = 0.52 p = 0.05). Negative associations were revealed between post-exercise T/C ratio and IGF-1 (R = - 0.58, p = 0.03) and serum free T and IL- ß (R = -0.56, p = 0.04) levels. The low level of pre-exercise genes and protein of the IL-1ß, IL-6 and TNF-α indicate a lack of inflammation signs in the female soccer players. This study shows significant effects of exercise on hormone levels and pro-inflammatory markers, which could be used to identify the role of female sex steroids on the immune function.

4.
Curr Pharm Biotechnol ; 21(12): 1213-1223, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32297577

RESUMO

BACKGROUND: The first immunosuppressive drug - cyclosporine A (CsA) has many unquestioned merits in maintaining organ transplants in patients, as well as, in the treatment of many inflammatory diseases, also associated with cutaneous manifestations. The main task of this drug is to suppress the inflammatory response at the sites of action, which is not well known. OBJECTIVE: The objective of this study was to evaluate the influence of CsA in therapeutic concentration on the expression of genes associated with the inflammatory response pathway in normal human dermal fibroblasts (NHDF; CC-2511), and this study attempted to determine the mechanism of its action. METHODS: The cytotoxicity MTT test was performed. The expression of the inflammatory response pathway genes was determined using HG-U133A_2.0 oligonucleotide microarrays. Statistical analysis was performed by GeneSpring 13.0 software using the PL-Grid platform. RESULTS: Among the 5,300 mRNA, only 573 were changed significantly in response to CsA compared to the control fibroblasts (P≤0.05). CsA inhibited the expression of most genes associated with the inflammatory response in NHDFs. There were only 19 genes with a fold change (FC) lower than -2.0, among which EGR1, FOS, PBK, CDK1 and TOP2A had the lowest expression, as did CXCL2 which can directly impact inflammation. Furthermore, ZNF451 was strongly induced, and COL1A1, COL3A1, IL33, TNFRSFs were weakly up-regulated (FC lower than 2.0). CONCLUSION: The CsA in therapeutic concentration influences the genes linked to the inflammatory response (in the transcriptional level) in human dermal fibroblasts. The findings suggest that the potential mechanism of CsA action in this concentration and on these genes can be associated with a profibrotic and proapoptotic, and genotoxic effects.


Assuntos
Ciclosporina/farmacologia , Fibroblastos/efeitos dos fármacos , Imunossupressores/farmacologia , Pele/efeitos dos fármacos , Transcriptoma , Técnicas de Cultura de Células , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/genética , Sobrevivência Celular/imunologia , Relação Dose-Resposta a Droga , Fibroblastos/imunologia , Perfilação da Expressão Gênica , Humanos , Inflamação/genética , Análise de Sequência com Séries de Oligonucleotídeos , RNA Mensageiro/genética , Pele/imunologia , Transcriptoma/efeitos dos fármacos , Transcriptoma/imunologia , Regulação para Cima
5.
Expert Rev Hematol ; 13(2): 117-126, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31971027

RESUMO

Introduction: The results of therapy of the multiple myeloma (MM) patients remain unsatisfactory despite the constantly observed progress in treatment.Areas covered: It has been shown that mechanisms regulated by heat shock proteins (HSPs) play an important role in pathogenesis of MM and resistance developing to treatment, which constitute a protective shield against external damaging factors in healthy and cancerous cells.Expert opinion: Inhibiting these mechanisms seems to be the natural way of therapy in MM patients. In vitro studies have shown promising effects in the form of an increase in the apoptosis index of MM cells exposed to HSP inhibitors. The observations are very promising in the early stages of clinical trials with HSP inhibitors, such as tanespimycin, in the relapsed/refractory MM patients. Effects were more pronounced when combined with bortezomib. It seems that enriching the range of anti-myeloma drugs with HSP inhibitors may be the next step in the future of extending life of patients with multiple myeloma.


Assuntos
Apoptose/efeitos dos fármacos , Benzoquinonas/uso terapêutico , Sistemas de Liberação de Medicamentos , Proteínas de Choque Térmico , Lactamas Macrocíclicas/uso terapêutico , Mieloma Múltiplo , Proteínas de Neoplasias , Proteínas de Choque Térmico/antagonistas & inibidores , Proteínas de Choque Térmico/metabolismo , Humanos , Mieloma Múltiplo/tratamento farmacológico , Mieloma Múltiplo/metabolismo , Mieloma Múltiplo/patologia , Proteínas de Neoplasias/antagonistas & inibidores , Proteínas de Neoplasias/metabolismo
6.
Wound Manag Prev ; 65(11): 19-32, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31702992

RESUMO

It remains unclear whether electrical currents can affect biological factors that determine chronic wound healing in humans. PURPOSE: The aim of this study was to determine whether anodal and cathodal high-voltage monophasic pulsed currents (HVMPC) provided to the area of a pressure injury (PI) change the blood level of cytokines (interleukin [IL]-1ß, IL-10, and tumor necrosis factor [TNF]-α) and growth factors (insulin-like growth factor [IGF]-1 and transforming growth factor [TGF]-ß1) in patients with neurological injuries and whether the level of circulatory cytokines and growth factors correlates with PI healing progression. METHODS: This study was part of a randomized clinical trial on the effects of HVMPC on PI healing. All patients with neurological injuries (spinal cord injury, ischemic stroke, and blunt trauma to the head) and a stage 2, stage 3, or stage 4 PI of at least 4 weeks' duration hospitalized in one rehabilitation center were eligible to participate if older than 18 years of age and willing to consent to donating blood samples. Exclusion criteria included local contraindications to electrical stimulation (cancer, electronic implants, osteomyelitis, tunneling, necrotic wounds), PIs requiring surgical intervention, patients with poorly controlled diabetes mellitus (HbA1C > 7%), critical wound infection, and/or allergies to standard wound treatment. Participants were randomly assigned to 1 of 3 groups: anodal (AG) or cathodal (CG) HVMPC treatment (154 µs; 100 Hz; 360 µC/sec; 1.08 C/day) or a placebo (PG, sham) applied for 50 minutes a day, 5 days per week, for 8 weeks. TNF-α, IL-1ß, IL-10, TGF-ß1, and IGF-1 levels in blood serum were assessed using the immunoenzyme method (ELISA) and by chemiluminescence, respectively, at baseline and week 4. Wound surface area measurements were obtained at baseline and week 4 and analyzed using a digitizer connected to a personal computer. Statistical analyses were performed using the maximum-likelihood chi-squared test, the analysis of variance Kruskal-Wallis test, the Kruskal-Wallis post-hoc test, and Spearman's rank order correlation; the level of significance was set at P ≤.05. RESULTS: Among the 43 participants, 15 were randomized to AG (mean age 53.87 ± 13.30 years), 13 to CG (mean age 51.08 ± 20.43 years), and 15 to PG treatment (mean age 51.20 ± 14.47 years). Most PIs were located in the sacral region (12, 74.42%) and were stage 3 (11, 67.44%). Wound surface area baseline size ranged from 1.00 cm2 to 58.04 cm2. At baseline, none of the variables were significantly different. After 4 weeks, the concentration of IL-10 decreased in all groups (AG: 9.8%, CG: 38.54%, PG: 27.42%), but the decrease was smaller in the AG than CG group (P = .0046). The ratio of pro-inflammatory IL-10 to anti-inflammatory TNF-α increased 27.29% in the AG and decreased 26.79% in the CG and 18.56% in the PG groups. Differences between AG and CG and AG and PG were significant (AG compared to CG, P = .0009; AG compared to PG, P = .0054). Other percentage changes in cytokine and growth factor concentration were not statistically significant between groups. In the AG, the decrease of TNF-α and IL-1ß concentrations correlated positively with the decrease of PI size (P <.05). CONCLUSION: Anodal HVMPC elevates IL-10/TNF-α in blood serum. The decrease of TNF-α and IL-1ß concentrations in blood serum correlates with a decrease of PI wound area. More research is needed to determine whether the changes induced by anodal HVMPC improve PI healing and to determine whether and how different electrical currents affect the activity of biological agents responsible for specific wound healing phases, both within wounds and in patients' blood. In clinical practice, anodal HVMPC should be used to increase the ratio of anti-inflammatory IL-10 to pro-inflammatory TNF-α , which may promote healing.


Assuntos
Citocinas/análise , Estimulação Elétrica/métodos , Peptídeos e Proteínas de Sinalização Intercelular/análise , Úlcera por Pressão/terapia , Traumatismos do Sistema Nervoso/sangue , Adulto , Idoso , Biomarcadores/análise , Biomarcadores/sangue , Citocinas/sangue , Estimulação Elétrica/instrumentação , Feminino , Humanos , Fator de Crescimento Insulin-Like I/análise , Peptídeos e Proteínas de Sinalização Intercelular/sangue , Interleucina-10/análise , Interleucina-10/sangue , Interleucina-1beta/análise , Interleucina-1beta/sangue , Masculino , Pessoa de Meia-Idade , Úlcera por Pressão/enzimologia , Estatísticas não Paramétricas , Fator de Crescimento Transformador beta1/análise , Fator de Crescimento Transformador beta1/sangue , Traumatismos do Sistema Nervoso/complicações , Traumatismos do Sistema Nervoso/fisiopatologia , Fator de Necrose Tumoral alfa/análise , Fator de Necrose Tumoral alfa/sangue
7.
Molecules ; 23(12)2018 Nov 28.
Artigo em Inglês | MEDLINE | ID: mdl-30487409

RESUMO

A new, simple, and cost-effective TLC-densitometric method has been established for the simultaneous quantitative determination of acetylsalicylic acid and ascorbic acid in combined effervescent tablets. Separation was performed on aluminum silica gel 60F254 plates using chloroform-ethanol-glacial acid at a volume ratio of 5:4:0.03 as the mobile phase. UV densitometry was performed in absorbance mode at 200 nm and 268 nm for acetylsalicylic acid and ascorbic acid, respectively. The presented method was validated as per ICH guidelines by specificity, linearity, accuracy, precision, limit of detection, limit of quantification, and robustness. Method validations indicate a good sensitivity with a low value of LOD and LOQ of both examined active substances. The linearity range was found to be 1.50⁻9.00 µg/spot and 1.50⁻13.50 µg/spot for acetylsalicylic and ascorbic acid, respectively. A coefficient of variation that was less than 3% confirms the satisfactory accuracy and precision of the proposed method. The results of the assay of combined tablet formulation equal 97.1% and 101.6% in relation to the label claim that acetylsalicylic acid and ascorbic acid fulfill pharmacopoeial requirements. The developed TLC-densitometric method can be suitable for the routine simultaneous analysis of acetylsalicylic acid and ascorbic acid in combined pharmaceutical formulations. The proposed TLC-densitometry may be an alternative method to the modern high-performance liquid chromatography in the quality control of above-mentioned substances, and it can be applied when HPLC or GC is not affordable in the laboratory.


Assuntos
Ácido Ascórbico/análise , Aspirina/análise , Densitometria/métodos , Sensibilidade e Especificidade
8.
Molecules ; 23(7)2018 07 05.
Artigo em Inglês | MEDLINE | ID: mdl-29976866

RESUMO

Effect of cyclosporin A (CsA) in a therapeutic concentration, on the expression of cytochrome P450 genes (CYPs), was investigated in normal human dermal fibroblast cells. The expression of 57 genes, encoding cytochrome P450 isoforms, was estimated using the microarray method. Amongst 396 normalized fluorescence signals related to cytochrome P450 activity, only 91 were strictly connected to CYPs and were analyzed using two methods: a self-organizing feature map of artificial neural networks and typical statistical analysis with significance level at p ≤ 0.05. Comparing the samples from fibroblasts cultured with CsA and those cultured without, up-regulated changes of CYP19A1, 1B1, 7A1, 7F1, 17A1 and down-regulated 2D6 gene expression were observed. The mRNAs with increased changes were in the same neuron of the self-organizing feature map. All distinguished CYPs encode monooxygenases, which plays an important role in steroids biosynthesis and metabolism. Based on the obtained results, we can conclude that CsA in therapeutic concentration changes the expression profile of CYPs in human dermal fibroblasts, especially affecting genes linked to steroids synthesis and/or metabolism. It shows the potential mechanism of action of CsA in human dermal fibroblast cells.


Assuntos
Ciclosporina/farmacologia , Sistema Enzimático do Citocromo P-450/genética , Fibroblastos/citologia , Perfilação da Expressão Gênica/métodos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Fibroblastos/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Redes Reguladoras de Genes , Humanos , Análise de Sequência com Séries de Oligonucleotídeos , Análise de Sequência de RNA , Esteroides/biossíntese , Esteroides/metabolismo
9.
Cancer Biomark ; 22(1): 89-99, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29562499

RESUMO

BACKGROUNDS: Colorectal cancer is the third most common cancer in economically developed countries. Molecular studies and, in particular, gene expression have contributed to advances in the diagnosis and treatment of many cancers. Genes can be molecular and therapeutic markers, but because of the large molecular diversity in colorectal cancer the knowledge is not yet fully established. Probably one of the most crucial processes during early cancer development is inflammation. The inflammatory response in the tumor is an important indicator of molecular etiology and later of cancer progression. OBJECTIVE: The aim of this work is to identify potential biomarkers for early stage of colorectal adenocarcinoma in patients' bowel tissues using transcriptomic analysis. METHODS: Expression of the inflammatory response genes of colorectal cancer at all clinical stages (I-IV) and control of the bowel were evaluated by oligonucleotide microarrays. RESULTS: Based on statistical analysis many differentially expressed genes were selected. LCK (LCK Proto-Oncogene, Src Family Tyrosine Kinase), GNLY (granulysin), SLC6A6 (Solute-Carrier Family 6 Member 6) and LAMP2 (Lysosomal Associated Membrane Protein 2) were specific for the early stage of the disease. These genes had the properties of the good biomarkers. CONCLUSIONS: The expression of LCK, GNLY, SLC6A6 and LAMP2 genes could be valuable potential diagnostic biomarkers of the early stage of colorectal adenocarcinoma.


Assuntos
Adenocarcinoma/diagnóstico , Biomarcadores Tumorais/sangue , Neoplasias Colorretais/diagnóstico , Detecção Precoce de Câncer/métodos , Perfilação da Expressão Gênica/métodos , Adenocarcinoma/genética , Adenocarcinoma/patologia , Neoplasias Colorretais/genética , Neoplasias Colorretais/patologia , Feminino , Humanos , Masculino , Estadiamento de Neoplasias , Proto-Oncogene Mas
10.
Adv Clin Exp Med ; 27(2): 207-216, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29521064

RESUMO

BACKGROUND: An integral part of the treatment of diabetes is physical activity. Scientific reports have shown the beneficial effects of hypoxia and exercise on cardiovascular and metabolic variables in patients with diabetes. OBJECTIVES: The aim of the study was to assess the effect of normobaric hypoxia and exercise on the serum concentrations of proangiogenic factors and glycemia in patients with type 1 diabetes. MATERIAL AND METHODS: A total of 28 adults (aged 30.4 years ±9.7 years), suffering from diabetes for 12.1 years ±6.0 years and healthy individuals, participated in the following trials: normoxic (Nor) and hypoxic (Hy) rest and Nor and Hy incremental exercise test (Ex) (FIO2 = 15.2%). The Altitude Trainer Hypoxico System (HYP-123 Hypoxic Generator, LOWOXYGEN Technology GmbH, Berlin, Germany) corresponding to a height of about 2500 m above sea level was used in the study. Exercise tests were performed on a cycle ergometer Excalibur Sport (Lode B.V., Groningen, The Netherlands). Cardiorespiratory variables, glycemia, angiogenic and hematological indices were measured at rest and in response to both exercise protocols. RESULTS: The present data confirmed that the patients with type 1 diabetes demonstrated a good level of aerobic capacity and fitness. NorEx and HyEx resulted in a significant decrease in serum glucose concentration (p < 0.05 vs p < 0.01). Patients with diabetes had higher baseline hypoxia induced factor-1alpha levels compared to healthy adults (p < 0.05), which increased after exposure to hypoxia and hypoxia with exercise (p < 0.001). Hypoxia significantly decreased baseline transforming growth factor-ß (TGF-ß) (p < 0.05) and had a significant effect on tumor necrosis factor-α level (TNF-α) (F = 4.9; p < 0.05). CONCLUSIONS: Our study demonstrated that hypoxia combined with exercise reduces glycemia and may induce significant benefits in the prevention of diabetes cardiovascular complications.


Assuntos
Diabetes Mellitus Tipo 1/terapia , Tolerância ao Exercício/fisiologia , Exercício Físico/fisiologia , Hipóxia/fisiopatologia , Adulto , Biomarcadores , Teste de Esforço , Alemanha , Humanos , Oxigênio/metabolismo , Aptidão Física
11.
Adv Clin Exp Med ; 26(6): 999-1004, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29068603

RESUMO

BACKGROUND: Regular and moderate exercise is beneficial for improving the efficiency of the heart, but high-intensity physical activity may result in cardiac changes. OBJECTIVES: This study focuses on the identification of the differences in echocardiography and blood variables before exercise, as well as the genes associated with cardiac hypertrophy at rest and in response to graded exercise test. MATERIAL AND METHODS: The study group was made up of 28 road cyclists. Echocardiographic parameters and blood pressure were measured before exercise tests (N = 28). Blood samples were collected at rest, at maximal exercise intensity in a graded bicycle test and after 15 min of recovery; afterwards, blood morphology was estimated and RNA was isolated. Analysis of the expression profile of genes was performed for randomly selected road cyclists using the microarray method. RESULTS: Echocardiographic results and blood parameters divided cyclists into two groups: with and without left ventricular hypertrophy (N = 14). Differences in the structure and function of the left ventricle cyclists with a similar level of training were observed (p < 0.05). Diastolic blood pressure and resting heart rate were significantly lower in subjects with left ventricular hypertrophy (p < 0.05). The myosin light chain 9 and interleukin-6 signal transducer gene expression were differentially regulated in cyclists with left ventricular hypertrophy compared to athletes with normal heart dimensions in response to intensive exercise. CONCLUSIONS: We have found differences in echocardiography parameters, blood pressure, stroke volume and maximal power in the cyclists examined. These studies indicate the benefits of the recommended echocardiography measurements for professional endurance-athletes. The graded exercise altered the myosin light chain 9 and interleukin-6 signal transducer gene expression in the peripheral blood of road cyclists has also been found.


Assuntos
Atletas , Ciclismo , Pressão Sanguínea/genética , Cardiomegalia Induzida por Exercícios/genética , Ecocardiografia , Hipertrofia Ventricular Esquerda/diagnóstico por imagem , Hipertrofia Ventricular Esquerda/genética , Resistência Física/genética , Volume Sistólico/genética , Adolescente , Adulto , Receptor gp130 de Citocina/genética , Teste de Esforço , Perfilação da Expressão Gênica , Genótipo , Humanos , Hipertrofia Ventricular Esquerda/fisiopatologia , Masculino , Cadeias Leves de Miosina/genética , Fenótipo , Transcriptoma , Função Ventricular Esquerda/genética , Adulto Jovem
12.
Acta Pol Pharm ; 73(1): 55-63, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27008801

RESUMO

Cyclosporin A is an immunosuppressant drug that is used not only in solid transplant rejection, but also in moderate and severe forms of psoriasis, pyoderma, lupus or arthritis. Serious side effects of the drug such as skin cancer or gingival hyperplasia probably start with the latent proliferation process. Little is known about the influence of cyclosporin A on molecular signaling in epidermal tissue. Thus, the aim of this study was to estimate the influence of cyclosporin A on the process of proliferation in normal human dermal fibroblasts. Fibroblasts were cultured in a liquid growth medium in standard conditions. Cyclosporin A was added to the culture after the confluence state. Survival and proliferation tests on human dermal fibroblast cells were performed. Total RNA was extracted from fibroblasts, based on which cDNA and cRNA were synthesized. The obtained cRNA was hybridized with the expression microarray HGU-133A_2.0. Statistical analysis of 2734 mRNAs was performed by the use of GeneSpring 13.0 software and only results with p < 0.05 were accepted. Analysis of variance with Tukey post hoc test with Benjamini-Hochberg correction for all three (8, 24, 48 h) culture stages (with and without cyclosporin A) was performed to lower the number of statistically significant results from 679 to 66, and less. Between statistically and biologically significant mRNAs down-regulated were EGRJ, BUBIB, MKI67, CDK1, TTK, E2F8, TPX2, however, the INSIG1, FOSL1, HMOX1 were up-regulated. The experiment data revealed that cyclosporin A up-regulated FOSL1 in the first 24 h, afterwards down-regulating its expression. The HMOX1 gene was up-regulated in the first stage of the experiment (CsA 8 h), however, after the next 16 h of culture time its expression was down-regulated (CsA 24 h), to finally increased in the later time period. The results indicate that cyclosporin A had a significant effect on proliferation in normal human dermal fibroblasts through the changes in the expression of genes related to the cell cycle and transcription regulation process.


Assuntos
Ciclosporina/efeitos adversos , Imunossupressores/efeitos adversos , Pele/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Fibroblastos/efeitos dos fármacos , Fibroblastos/fisiologia , Heme Oxigenase-1/genética , Humanos , Pele/patologia
13.
J Hum Kinet ; 40: 103-11, 2014 Mar 27.
Artigo em Inglês | MEDLINE | ID: mdl-25031678

RESUMO

The aim of this study was to evaluate adrenergic responses in the peripheral blood of trained road cyclists at rest, at maximal intensity of incremental bicycle exercise test, and during 15 minutes of recovery, as well as the relationship between them. Competitive male road cyclists, in the pre-competitive phase of a season, mean age 21.7 ± 6.4 years, and BMI 20.7 ± 0.8 kg·m(-2), performed an incremental test on a bicycle ergometer with unloaded cycling for 5 min, then increased the load to 40 W every 3 min, up to maximal exercise intensity. The plasma catecholamine concentrations (epinephrine, norepinephrine) and oxygen uptake were estimated. The expression of 132 genes related to the adrenergic system in leukocytes was measured. A statistically significant increase in plasma epinephrine concentration (p < 0.01) was observed in response to exercise. The mean of maximal oxygen uptake was 65.7 ± 5.5 ml·kg(-1)·min(-1). The RGS2 gene expression was highest regardless of the test phase for all athletes. The effort had a statistically significant influence on ADRB2 and RAB2A expression. In addition, the RAB2A, ADM and HSPB1 expression level increased during recovery. We can conclude that plasma epinephrine concentration and genes related to the adrenergic system such as ADM, ADRB2, CCL3, GPRASP1, HSPB1, RAB2A, RGS2 and ROCK1 seem to have an influence on the response to high-intensity exercise in trained cyclists.

14.
Ginekol Pol ; 85(3): 172-9, 2014 Mar.
Artigo em Polonês | MEDLINE | ID: mdl-24783427

RESUMO

INTRODUCTION: Influence of histamine on tumor development remains obscure. The exact mechanism of this action is not known. Different data indicate high concentrations of histamine in tumor tissues, such as malignant melanoma, breast cancer, colon carcinoma, lymphomas and leukemia. To the best of our knowledge, the literature offers no reports about the role of histamine and of differences between expression patterns of histamine-related genes in endometrial cancer AIM: The aim of the study was a comparative analysis of the gene expression profile involved with histaminergic system in endometrioid endometrial cancer in relation to histologically normal endometrium, and identification of differentiation genes whose transcriptional activity significantly differs in pathomorphological grades G1,G2, G3 of endometrial cancer as compared to the control group. MATERIAL AND METHODS: Total RNA was extracted from 24 endometrial probes using TRIzol reagent (Invitrogen). The expression profile of 119 transcripts associated with histaminergic system was assessed using oligonucleotide microarrays of HG-U 133A (Affymetrix). After normalization of the results with RMA Express software, differentiation genes were mined by the use of one-way analysis ANOVA and U Mann-Whitney test carried out in Gene Spring 11.5 software. RESULTS: Among 119 transcripts, 14 expressed more than 1.5-fold change and were significant at p<0.05 in endometrioid endometrial cancer in relation to the normal endometrium. Further analysis led to the identification of differentially expressed genes in grades G1, G2 and G3 of endometrial adenocarcinoma as compared to the control group, which were specific for each of the studied groups in grade G1 (CPA3), in grade G2 (HNMT LYN, DPT ITPKB, RASA4, APR RAB1 1FIP1, YWHAZ, VAMP8, RAB25) and in grade G3 (HRH3). CONCLUSIONS: Our results confirmed the role of the histaminergic system in the pathogenesis of endometrial adenocarcinoma. The observed differences in the expression of those genes, depending on the grade of adenocarcinoma, may indicate an important role of the isolated differentiation genes in endometrial tumorigenesis.


Assuntos
Adenocarcinoma/genética , Carcinogênese/genética , Neoplasias do Endométrio/genética , Neoplasias do Endométrio/metabolismo , Endométrio/metabolismo , Regulação Neoplásica da Expressão Gênica/genética , Histamina/genética , Adenocarcinoma/metabolismo , Adenocarcinoma/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Neoplasias do Endométrio/patologia , Feminino , Perfilação da Expressão Gênica , Genes Neoplásicos/genética , Histamina/metabolismo , Humanos , Pessoa de Meia-Idade , Gradação de Tumores , Análise de Sequência com Séries de Oligonucleotídeos
15.
Am J Hum Biol ; 24(6): 859-62, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22915245

RESUMO

OBJECTIVE: The present study has focused on the identification of differences between the expression pattern of TGF-ß signaling pathway genes in athletes after exercise. MATERIALS AND METHODS: The study group consisted of three healthy cyclists, which were collected pre-exercise, immediately postexercise, and after 15 min of recovery. The analysis of the expression profile of genes related to the TGF-ß signal transduction pathway was performed in peripheral blood mononuclear cells using HG-U133A oligonucleotide microarrays. RESULTS: A significant differential gene expression was recorded for RUNX3, TGFBR3, MLC1, and GRB2. CONCLUSIONS: The effect of physical exercise on immune response may be essential for human health. Moreover, alterations of TGF-ß signaling can be involved in the process of adaptation of human organism to physical exertion.


Assuntos
Exercício Físico , Leucócitos Mononucleares/metabolismo , Transcriptoma , Fator de Crescimento Transformador beta/genética , Fator de Crescimento Transformador beta/imunologia , Adulto , Ciclismo/fisiologia , Perfilação da Expressão Gênica , Humanos , Leucócitos Mononucleares/imunologia , Masculino , Análise de Sequência com Séries de Oligonucleotídeos , Esforço Físico , Transdução de Sinais , Fator de Crescimento Transformador beta/metabolismo , Adulto Jovem
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