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1.
Sci Rep ; 11(1): 3122, 2021 02 04.
Artigo em Inglês | MEDLINE | ID: mdl-33542424

RESUMO

Sample pooling strategy was intended to determine the optimal parameters for group testing of pooled specimens for the detection of SARS-CoV-2 and process them without significant loss of test usability. Standard molecular diagnostic laboratory equipment, and commercially available centrifugal filters, RNA isolation kits and SARS Cov2 PCR tests were used. The basic idea was to combine and concentrate several samples to the maximal volume, which can be extracted with the single extraction column. Out of 16 tested pools, 12 were positive with cycle threshold (Ct) values within 0.5 and 3.01 Ct of the original individual specimens. The analysis of 112 specimens determined that 12 pools were positive, followed by identification of 6 positive individual specimens among the 112 tested. This testing was accomplished with the use of 16 extractions/PCR tests, resulting in saving of 96 reactions but adding the 40 centrifugal filters. The present study demonstrated that pool testing could detect even up to a single positive sample with Ct value as high as 34. According to the standard protocols, reagents and equipment, this pooling method can be applied easily in current clinical testing laboratories.


Assuntos
Teste de Ácido Nucleico para COVID-19/métodos , COVID-19/diagnóstico , RNA Viral/isolamento & purificação , Manejo de Espécimes/métodos , Humanos , SARS-CoV-2/genética , SARS-CoV-2/isolamento & purificação , Sensibilidade e Especificidade
2.
Food Chem Toxicol ; 109(Pt 2): 1026-1031, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-28487231

RESUMO

The dichloromethane extract from fruits of Angelica archangelica L. was separated by the modern high-performance countercurrent chromatography (HPCCC). The extract and five pure compounds: xanthotoxin, bergapten, imperatorin, phellopterin and isoimperatorin, and the mixture of imperatorin and phellopterin, have been studied as the potential antiviral agents against Herpes simplex virus type l and Coxsackievirus B3. The cytotoxicity was measured using the MTT method. Compounds were tested for the in vitro antiviral activity using the cytopathic effect (CPE) inhibitory assay and by the virus titre reduction assay. Real-time PCR was used to quantify the relative inhibition of the HSV-1 replication. The results indicate that the highest activity was demonstrated by the extract, imperatorin, phellopterin and the mixture of imperatorin and phellopterin, reducing the HSV-1 replication by 5.61 log, 4.7 log, 3.01 log and 3.73 log, respectively. The influence of isolated compounds on the CVB3 replication was not significant. Only the extract caused the decrease in the titre of virus in relation to the virus control. Our results show that coumarins of A. archangelica L. might be a potential candidate for the development of the alternative natural anti- HSV-1 compound. Moreover, the presence of isopentenyloxy moiety at C-8 position significantly improves their activity.


Assuntos
Angelica archangelica/química , Antivirais/química , Antivirais/farmacologia , Enterovirus Humano B/efeitos dos fármacos , Herpesvirus Humano 1/efeitos dos fármacos , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Antivirais/isolamento & purificação , Distribuição Contracorrente , Infecções por Coxsackievirus/virologia , Enterovirus Humano B/fisiologia , Herpes Simples/virologia , Herpesvirus Humano 1/fisiologia , Humanos , Extratos Vegetais/isolamento & purificação , Replicação Viral/efeitos dos fármacos
3.
Infect Agent Cancer ; 11: 45, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27547238

RESUMO

BACKGROUND: Each year approximately 6,000 new cases of head and neck cancer are registered in Poland. Human papillomavirus (HPV) and Epstein-Barr virus (EBV) have been associated with tumour formation. Cytokines have been shown to play an important role both in inflammation and carcinogenesis and they can be detected in saliva and serum with ELISA assays. Salivary biomarkers may be used as markers of early cancer detection. The aim of this study was the analysis of the serum and salivary levels of IL-10, TNF-α, TGF-ß and VEGF in patients with oropharyngeal cancer and in healthy individuals. The level of these biomarkers was also analyzed in HPV- and EBV-related cases. METHODS: The study involved 78 patients with histopathologically confirmed oropharyngeal squamous cell carcinoma and 40 healthy controls. Serum and salivary levels of IL-10, TNF-α, TGF-ß and VEGF were analyzed both in patients and in healthy individuals by ELISA method using Diaclone SAS commercially available kits (France). EBV DNA was detected by the nested PCR for amplification of EBNA-2. HPV detection and genotyping was performed using the INNO-LiPA HPV Genotyping Extraassay (Innogenetics N. V, Gent, Belgium). The obtained results were subjected to statistical analysis using Mann-Whitney and Kruskal Wallis tests. Test values of p < 0.05 were considered statistically significant. RESULTS: The level of tested cytokines was higher in patients than in controls both in serum (IL-10: 2.3 pg/ml vs 1.65 pg/ml, p = 0.0003; TGF-ß: 11.3 ng/ml vs 7.8 ng/ml, p = 0.0005; VEGF: 614 pg/ml vs 210 pg/ml, p = 0.0004; TNF-α: 15.0 ng/ml vs 12.90 ng/ml, p = 0.1397) as well as in saliva (IL-10: 5.9 pg/ml vs 2.5 pg/ml, p = 0.00002; TGF-ß: 24.1 ng/ml vs 14.8 ng/ml, p = 0.00002; VEGF: 4321 pg/ml vs 280 pg/ml, p = 0.0000; TNF-α: 23.1 ng/ml vs 11.3 ng/ml, p = 0.00002). EBV DNA was detected in 51.3 % of patients and 20 % of controls, HPV DNA was present in 30.8 % of patients and 2,5 % of controls. The level of IL-10 was statistically higher in patients infected with EBV, HPV and co-infected with EBV/HPV. The level of TNF-α was significantly higher in patients infected with EBV, while TGF-ß in patients with HPV infection and EBV/HPV co-infection. CONCLUSION: Detection of salivary cytokines may be very helpful in early diagnosis, treatment and prognosis of OSCC.

4.
Infect Agent Cancer ; 10: 37, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26462255

RESUMO

OBJECTIVE: The aim of this study was to analyze the prevalence of HPV and EBV in oral and oropharyngeal squamous cell carcinoma in south-eastern Poland. The correlation between viral infection, OSCC, alcohol use, tobacco smoking, demographic data (gender, age, place of residence), anatomic location, pre-treatment staging, evidence of metastases to lymph nodes, and grading was also investigated. METHODS: The examination samples were collected from paraffin tissue blocks, from 154 patients. Viral DNA was amplified by the nested-PCR method. RESULTS: HPV DNA was detected in 29.2 % of the tested samples (in 27.4 % of oropharyngeal and in 30.4 % of oral cavity). The HPV type 16 was detected in 15.6 % of all samples, and in 53.3 % of HPV-positive group. In HPV-positive samples from oropharyngeal HPV 16 constitute 76.5 %, and in HPV-positive samples from oral cavity HPV 16 constitute 39.3 %. Mixed infection (more than one type of HPV) was observed in 23.5 and 60.7 %, respectively, and in 46.7 % of all HPV-positive samples, and in 12.3 % of the whole study group. EBV DNA was detected in 27.3 % of the cases and HPV-EBV co-infection in 7.8 % of samples. CONCLUSIONS: In major patients from Southeastern region of Poland with oropharyngeal cancer HPV type 16 was detected but in oral cavity cancer other mixed infections were observed (i.e. 51, 52, 59, 66, 68, 71, 74). HPV 16 was detected more often among patients younger than 50 years of age, whereas the mixed HPV in the group aged 50-59. The pathogenesis of oral squamous cell carcinoma may be connected with EBV infection. Future studies on the mechanisms of HPV/EBV co-infection and/or superinfection and their role in oral squamous cell carcinoma are necessary.

5.
Anticancer Res ; 35(3): 1657-61, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25750324

RESUMO

The role of viruses in the etiology of oral cancer has been proposed in many studies. The aim of the present study was to analyze the prevalence of Epstein-Barr virus, Human Herpes virus type 1, Cytomegalovirus and Human Papilloma virus among patients with oral squamous cell carcinoma in a Polish population. We investigated fresh-frozen tumor tissue fragments obtained from 80 patients with OSCC using the polymerase chain reaction assay. HPV was detected in 32.5% (22.5% were HPV 16), more often in laryngeal (36%) than in oropharyngeal carcinoma (26.6%). EBV was identified in 57.5%, HHV-1 in 7.5%, and CMV in 10% of patients. Co-infection with one or more viruses was detected in 30% of cases and most frequently it was co-infection with EBV and HPV (15%). Further studies are necessary to determine the potential role of EBV and the possible importance of HHV-1 as an infection co-factor in oropharyngeal cancer.


Assuntos
Carcinoma de Células Escamosas/virologia , Citomegalovirus/isolamento & purificação , Herpesvirus Humano 4/isolamento & purificação , Neoplasias Laríngeas/virologia , Neoplasias Orofaríngeas/virologia , Papillomaviridae/isolamento & purificação , Simplexvirus/isolamento & purificação , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
6.
Ann Agric Environ Med ; 22(1): 106-9, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25780838

RESUMO

INTRODUCTION: Head and neck cancers are the most common cancers worldwide. It is estimated that approximately 90% of all head and neck cancers are represented by squamous cell carcinoma (SCC). There are many risk factors causing this type of cancer, including environmental factors and lifestyle choices, such as tobacco smoking or abusing alcohol. Other important risk factor include infectious factors. OBJECTIVE: The aim of this study was to analyze the prevalence of BK and JC virus infections among patients with oral squamous cell carcinoma (OSCC). MATERIALS AND METHOD: The correlation between BKV infection and OSCC, and correlation between BKV, OSCC, alcohol abuse, tobacco smoking, demographic data, pre-treatment staging, metastases of lymph node evidence, and grading, was analyzed. The study group consisted of 92 patients with squamous cell carcinoma (OSCC), 75 males, and 17 females, aged between 40 - 87 (average 56.8). All the patients underwent surgery and were not subjected to chemotherapy or radiotherapy prior to treatment. The analyzed samples were collected from paraffin sections. RESULTS: BKV DNA was detected in 18.5% of patients with OSCC. In the control group, BKV DNA was detected in 3.3%. BKV DNA was statistically more frequently detected among patients with squamous carcinoma, compared to the control group (p<0.05). CONCLUSIONS: The obtained results suggest that the BKV virus may play an important role in the development of oral squamous cell carcinoma.


Assuntos
Vírus BK/isolamento & purificação , Carcinoma de Células Escamosas/epidemiologia , Vírus JC/isolamento & purificação , Neoplasias Bucais/epidemiologia , Infecções por Polyomavirus/epidemiologia , Infecções Tumorais por Vírus/epidemiologia , Adulto , Idoso de 80 Anos ou mais , Carcinoma de Células Escamosas/complicações , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias Bucais/complicações , Polônia/epidemiologia , Infecções por Polyomavirus/complicações , Prevalência , Fatores de Risco , Infecções Tumorais por Vírus/complicações
7.
Stem Cell Reports ; 3(6): 987-99, 2014 Dec 09.
Artigo em Inglês | MEDLINE | ID: mdl-25454634

RESUMO

Inducing organogenesis in 3D culture is an important aspect of stem cell research. Anterior neural structures have been produced from large embryonic stem cell (ESC) aggregates, but the steps involved in patterning such complex structures have been ill defined, as embryoid bodies typically contained many cell types. Here we show that single mouse ESCs directly embedded in Matrigel or defined synthetic matrices under neural induction conditions can clonally form neuroepithelial cysts containing a single lumen in 3D. Untreated cysts were uniformly dorsal and could be ventralized to floor plate (FP). Retinoic acid posteriorized cysts to cervical levels and induced localize FP formation yielding full patterning along the dorsal/ventral (DV) axis. Correct spatial organization of motor neurons, interneurons, and dorsal interneurons along the DV axis was observed. This system serves as a valuable tool for studying morphogen action in 3D and as a source of patterned spinal cord tissue.


Assuntos
Células-Tronco Embrionárias/citologia , Tubo Neural/citologia , Tubo Neural/fisiologia , Organogênese , Animais , Biomarcadores , Padronização Corporal/efeitos dos fármacos , Padronização Corporal/genética , Diferenciação Celular/efeitos dos fármacos , Desenvolvimento Embrionário/efeitos dos fármacos , Desenvolvimento Embrionário/genética , Células-Tronco Embrionárias/efeitos dos fármacos , Células-Tronco Embrionárias/metabolismo , Expressão Gênica , Proteínas Hedgehog/genética , Camundongos , Organogênese/efeitos dos fármacos , Organogênese/genética , Técnicas de Cultura de Tecidos , Tretinoína/farmacologia
8.
Pol J Microbiol ; 63(1): 105-9, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25033670

RESUMO

The aim of this study was to compare the prevalence of EBV genotype and del-LMP-1 in saliva from Polish, Taiwanese and Arabic healthy students. The study group consisted of 56 healthy students; 24 of them Polish, 25 Taiwanese, and 7 Arabic. Typing was carried out using PCR with EBNA-2 primers. A detection of LMP-1 variants was also performed using PCR. EBV DNA was detected in 22 investigated samples (39.3%). Type 1 of the virus was dominant in both Polish and Taiwanese group. Among 62.5% Taiwanese with EBV 1 and 55.6% Polish detected EBV with 30-bp deletion in LMP-1 gene.


Assuntos
Genótipo , Herpesvirus Humano 4/genética , Filogenia , Proteínas da Matriz Viral/metabolismo , Adolescente , Adulto , Portador Sadio/virologia , Feminino , Regulação Viral da Expressão Gênica/fisiologia , Variação Genética , Herpesvirus Humano 4/isolamento & purificação , Humanos , Masculino , Polônia , Arábia Saudita , Taiwan , Proteínas da Matriz Viral/genética , Adulto Jovem
9.
EMBO J ; 28(5): 591-601, 2009 Mar 04.
Artigo em Inglês | MEDLINE | ID: mdl-19165148

RESUMO

The ultraviolet-B (UV-B) portion of the solar radiation functions as an environmental signal for which plants have evolved specific and sensitive UV-B perception systems. The UV-B-specific UV RESPONSE LOCUS 8 (UVR8) and the multifunctional E3 ubiquitin ligase CONSTITUTIVELY PHOTOMORPHOGENIC 1 (COP1) are key regulators of the UV-B response. We show here that uvr8-null mutants are deficient in UV-B-induced photomorphogenesis and hypersensitive to UV-B stress, whereas overexpression of UVR8 results in enhanced UV-B photomorphogenesis, acclimation and tolerance to UV-B stress. By using sun simulators, we provide evidence at the physiological level that UV-B acclimation mediated by the UV-B-specific photoregulatory pathway is indeed required for survival in sunlight. At the molecular level, we demonstrate that the wild type but not the mutant UVR8 and COP1 proteins directly interact in a UV-B-dependent, rapid manner in planta. These data collectively suggest that UV-B-specific interaction of COP1 and UVR8 in the nucleus is a very early step in signalling and responsible for the plant's coordinated response to UV-B ensuring UV-B acclimation and protection in the natural environment.


Assuntos
Proteínas de Arabidopsis/fisiologia , Arabidopsis/fisiologia , Proteínas Cromossômicas não Histona/fisiologia , Raios Ultravioleta , Aclimatação , Proteínas de Arabidopsis/genética , Proteínas Cromossômicas não Histona/genética , Regulação da Expressão Gênica de Plantas , Hipocótilo/crescimento & desenvolvimento , Hipocótilo/fisiologia , Mutação , Ligação Proteica , Transdução de Sinais/fisiologia , Estresse Fisiológico , Luz Solar , Ubiquitina-Proteína Ligases
10.
Plant J ; 54(3): 402-14, 2008 May.
Artigo em Inglês | MEDLINE | ID: mdl-18266923

RESUMO

Ultraviolet-B light (UV-B) regulates the expression of genes in a wavelength- and fluence rate-dependent fashion. A signaling pathway consisting of CONSTITUTIVE PHOTOMORPHOGENESIS 1 (COP1) and UV RESISTANCE LOCUS 8 (UVR 8) mediates responsiveness to longer wavelength, low intensity UV-B light-activating, for example, HY5 gene expression. By contrast, transcription of another group of genes, including ANAC13, modulated by shorter wavelength, higher intensity UV-B is controlled by a yet unknown and largely COP1-independent signaling cascade. Here we provide evidence by promoter deletion analysis, and characterization of genetic mutants displaying aberrant expression patterns, that two cis-regulatory elements, designated MRE(ANAC13) and UVBox(ANAC13), are required for maximal UV-B induction of the ANAC13 gene in transgenic plants. These elements are located in the proximal 150-bp region of the ANAC13 promoter. They show no significant similarity to each other; the putative MRE(ANAC13) (-AACCTT-) is closely related to MRE(CHS) (-AACCTA-) found in the CHALCONE SYNTHASE (CHS) gene, whereas UVBox(ANAC13) (with core sequence CAAG) represents a novel cis-regulatory element. The novel UVBox(ANAC13) sequence is significantly enriched in the promoter region of a subset of UV-B-induced genes with similar activation properties as ANAC13. In addition, we demonstrate that expression of a chimeric gene containing only the dimerized 12-mer containing UVBox(ANAC13) fused to a minimal CaMV35S promoter/luciferase reporter is (i) efficiently induced by shorter wavelength, higher intensity UV-B, but (ii) does not respond either to longer wavelength UV-B and red light or (iii) to abscisic acid treatment and osmotic, salt, heat and cold stresses.


Assuntos
Arabidopsis/genética , Elementos Reguladores de Transcrição/genética , Transcrição Genética/efeitos da radiação , Raios Ultravioleta , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Sequência de Bases , Luz , Dados de Sequência Molecular , Mutação Puntual , Regiões Promotoras Genéticas/genética , Ubiquitina-Proteína Ligases
11.
Przegl Epidemiol ; 56(1): 65-72, 2002.
Artigo em Polonês | MEDLINE | ID: mdl-12150069

RESUMO

CMV and EBV infections are common in humans. In immunocompetent persons those infections are usually asymptomatic but in immunocompromised can manifest as a severe disease. CMV is a common cause of congenital infections. It is also a frequent complication in transplant recipients. The aim of this study was to assess the prevalence of CMV and EBV infections among hospitalized children. Specific antibodies against CMV and EBV were detected in serum by ELISA test. Presence of CMV DNA was determined in leucocytes by Murex Hybrid Capture System. CMV and EBV infections were defined as the presence in serum IgM-class specific antibodies. Obtained results indicate that CMV and EBV infections are frequent in immunocompromised patients. Among patients with CMV or EBV infection, 40% have been diagnosed with cancer, most of whom with hematologic malignancies: leukemia or lymphoma. CMV and EBV coinfection was detected in 14% of infected children. Of all patients with CMV, 50% were neonates and infants. Congenital infection was diagnosed only in one case. The remaining infections were acquired during perinatal period or later.


Assuntos
Infecções por Citomegalovirus/epidemiologia , Citomegalovirus/isolamento & purificação , Infecções por Vírus Epstein-Barr/epidemiologia , Herpesvirus Humano 4/isolamento & purificação , Pacientes Internados/estatística & dados numéricos , Adolescente , Adulto , Fatores Etários , Anticorpos Antivirais/sangue , Criança , Pré-Escolar , Citomegalovirus/imunologia , Infecções por Citomegalovirus/diagnóstico , Infecções por Citomegalovirus/imunologia , Ensaio de Imunoadsorção Enzimática , Infecções por Vírus Epstein-Barr/diagnóstico , Infecções por Vírus Epstein-Barr/imunologia , Feminino , Herpesvirus Humano 4/imunologia , Humanos , Hospedeiro Imunocomprometido , Lactente , Recém-Nascido , Masculino , Neoplasias/complicações , Neoplasias/virologia , Polônia/epidemiologia , Prevalência
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