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1.
Artículo en Ruso | MEDLINE | ID: mdl-38549413

RESUMEN

Anterior cervical osteophytes are a fairly common X-ray finding in people over 50 years old. Incidence of dysphagia in patients with anterior osteophytes varies from 1% in those aged 40-60 years to 10.6% in patients over 60 years old. The most common causes of anterior cervical hyperosteophytosis causing dysphagia are cervical spondylosis deformans and Forestier disease. We present 2 clinical cases of spondylogenic dysphagia in cervical spondylosis deformans and Forestier disease. The review is devoted to the causes and diagnostic methods for dysphagia caused by anterior cervical osteophytes, as well as surgical options for this pathology. CONCLUSION: Microsurgical resection of anterior osteophytes is an effective method for dysphagia after ineffective therapy for 3 months. Microsurgical osteophytectomy provides stable regression of dysphagia with low recurrence rate.


Asunto(s)
Trastornos de Deglución , Hiperostosis Esquelética Difusa Idiopática , Osteofito , Espondilosis , Humanos , Persona de Mediana Edad , Hiperostosis Esquelética Difusa Idiopática/complicaciones , Hiperostosis Esquelética Difusa Idiopática/diagnóstico , Hiperostosis Esquelética Difusa Idiopática/cirugía , Osteofito/complicaciones , Osteofito/diagnóstico por imagen , Osteofito/cirugía , Trastornos de Deglución/diagnóstico por imagen , Trastornos de Deglución/etiología , Trastornos de Deglución/cirugía , Espondilosis/complicaciones , Espondilosis/diagnóstico por imagen , Espondilosis/cirugía , Vértebras Cervicales/diagnóstico por imagen , Vértebras Cervicales/cirugía , Vértebras Cervicales/patología
2.
Vestn Ross Akad Med Nauk ; (2): 35-41, 2012.
Artículo en Ruso | MEDLINE | ID: mdl-22642176

RESUMEN

Two approaches to somatic point mutations in 12 and 13 codones of K-ras gene were analyzed: PCR/SSCP/ACRS/sequencing and allele-specific PCR in the real-life regimen (Russian set "KRAS-7M"). The comparison was carried out on 62 examples of genomic DNA extracted from frozen colon carcinomas, which underwent manual dissection. The results obtained in two attempts were consistent in 95,2% (N=59). Specificity and sensitivity of K-ras mutations detection using "KRAS-7M" set were 100 and 96,4% respectively, and 94,1 and 100% respectievly using PCR/SSCP/ACRS/automatic sequencing. False positive results were absent when detecting with "KRAS-7M" and accounted for 2 cases (5,9%) when using PCR/SSCP/ ACRS/automatic sequencing. The only false negative response (3,6%) was obtained analyzing mutations using "KRAS-7M".


Asunto(s)
Neoplasias Colorrectales/genética , Análisis Mutacional de ADN , ADN de Neoplasias/análisis , Genes ras , Técnicas de Diagnóstico Molecular , Mutación Puntual , Adulto , Anciano , Anciano de 80 o más Años , Neoplasias Colorrectales/patología , Análisis Mutacional de ADN/métodos , Análisis Mutacional de ADN/normas , Femenino , Pruebas Genéticas/métodos , Pruebas Genéticas/normas , Humanos , Masculino , Persona de Mediana Edad , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Diagnóstico Molecular/normas , Polimorfismo Conformacional Retorcido-Simple , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Reacción en Cadena en Tiempo Real de la Polimerasa/normas , Estudios Retrospectivos , Sensibilidad y Especificidad
3.
Vavilovskii Zhurnal Genet Selektsii ; 26(5): 413-421, 2022 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-36128569

RESUMEN

Wheatgrass (Th. intermedium) has been traditionally used in wheat breeding for obtaining wheat-wheatgrass hybrids and varieties with introgressions of new genes for economically valuable traits. However, in the 1980s in the United States wheatgrass was selected from among perennial plant species as having promise for domestication and the development of dual-purpose varieties for grain (as an alternative to perennial wheat) and hay. The result of this work was the creation of the wheatgrass varieties Kernza (The Land Institute, Kansas) and MN-Clearwater (University of Minnesota, Minnesota). In Omsk State Agrarian University, the variety Sova was developed by mass selection of the most winter-hardy biotypes with their subsequent combination from the population of wheatgrass obtained from The Land Institute. The average grain yield of the variety Sova is 9.2 dt/ha, green mass is 210.0 dt/ ha, and hay is 71.0 dt/ha. Wheatgrass is a crop with a large production potential, benef icial environmental properties, and valuable grain for functional food. Many publications show the advantages of growing the Kernza variety compared to annual crops in reducing groundwater nitrate contamination, increasing soil carbon sequestration, and reducing energy and economic costs. However, breeding programs for domestication of perennial crops are very limited in Russia. This paper presents an overview of main tasks faced by breeders, aimed at enhancing the yield and cultivating wheatgrass eff iciency as a perennial grain and fodder crop. To address them, both traditional and modern biotechnological and molecular cytogenetic approaches are used. The most important task is to transfer target genes of Th. intermedium to modern wheat varieties and decrease the level of chromatin carrying undesirable genes of the wild relative. The f irst consensus map of wheatgrass containing 10,029 markers was obtained, which is important for searching for genes and their introgressions to the wheat genome. The results of research on the nutritional and technological properties of wheatgrass grain for the development of food products as well as the differences in the quality of wheatgrass grain and wheat grain are presented.

4.
Vavilovskii Zhurnal Genet Selektsii ; 25(7): 740-745, 2021 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-34950845

RESUMEN

Present-day wheat breeding for immunity exploits extensively closely related species from the family Triticeae as gene donors. The 2NS/2AS translocation has been introduced into the genome of the cultivated cereal Triticum aestivum from the wild relative T. ventricosum. It contains the Lr37, Yr17, and Sr38 genes, which support seedling resistance to the pathogens Puccinia triticina Eriks., P. striiformis West. f. sp. tritici, and P. graminis Pers. f. sp. tritici Eriks. & E. Henn, which cause brown, yellow, and stem rust of wheat, respectively. This translocation is present in the varieties Trident, Madsen, and Rendezvous grown worldwide and in the Russian varieties Morozko, Svarog, Graf, Marquis, and Homer bred in southern regions. However, the Sr38 gene has not yet been introduced into commercial varieties in West Siberia; thus, it remains of practical importance for breeding in areas where populations of P. graminis f. sp. tritici are represented by avirulent clones. The main goal of this work was to analyze the frequency of clones (a)virulent to the Sr38 gene in an extended West Siberian collection of stem rust agent isolates. In 2019-2020, 139 single pustule isolates of P. graminis f. sp. tritici were obtained on seedlings of the standard susceptible cultivar Khakasskaya in an environmentally controlled laboratory (Institute of Cytology and Genetics SB RAS) from samples of urediniospores collected on commercial and experimental bread wheat f ields in the Novosibirsk, Omsk, Altai, and Krasnoyarsk regions. By inoculating test wheat genotypes carrying Sr38 (VPM1 and Trident), variations in the purity of (a)virulent clones were detected in geographical samples of P. graminis f. sp. tritici. In general, clones avirulent to Sr38 constitute 60 % of the West Siberian fungus population, whereas not a single virulent isolate was detected in the Krasnoyarsk collection. The Russian breeding material was screened for sources of the stem rust resistance gene by using molecular markers specif ic to the 2NS/2AS translocation. A collection of hybrid lines and varieties of bread spring wheat adapted to West Siberia (Omsk SAU) was analyzed to identify accessions promising for the region. The presence of the gene was postulated by genotyping with specif ic primers (VENTRIUP-LN2) and phytopathological tests with avirulent clones of the fungus. Dominant Sr38 alleles were identif ied in Lutescens 12-18, Lutescens 81-17, Lutescens 66-16, Erythrospermum 79/07, 9-31, and 8-26. On the grounds of the composition of the West Siberian P. graminis f. sp. tritici population, the Sr38 gene can be considered a candidate for pyramiding genotypes promising for the Novosibirsk, Altai, and Krasnoyarsk regions.

5.
Vavilovskii Zhurnal Genet Selektsii ; 25(5): 543-551, 2021 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-34595376

RESUMEN

Spring bread wheat is the staple crop in Western Siberia and Kazakhstan, a signif icant portion of which goes for export. Wheat breeding with a high level of zinc in wheat grain is the most cost-effective and environmentally friendly way to address zinc def iciency in the diet. The purpose of this work was to evaluate the contribution of the factors 'location' and 'genotype' in the variability of zinc content in wheat grain, and to identify the best varieties as sources of this trait for breeding. The research on screening zinc content in the wheat grain of 49 spring bread wheat varieties from the Kazakhstan- Siberia Spring Wheat Trial (KASIB) nursery was carried out at 4 sites in Russia (Chelyabinsk, Omsk, Tyumen, Novosibirsk) and 2 sites in Kazakhstan (Karabalyk and Shortandy) in 2017-2018. The content of zinc in wheat grain was evaluated at the Ionomic Facility of University of Nottingham in the framework of the EU project European Plant Phenotyping Network-2020. The analysis of variance showed that the main contribution into the general phenotypic variation of the studied trait, 38.7 %, was made by the factor 'location' due to different contents of zinc and moisture in the soil of trial sites; the effect of the factor 'year' was 13.5 %, and the effect of the factor 'genotype' was 8.0 %. The most favorable environmental conditions for accumulation of zinc in wheat grain were observed in the Omsk region. In Omsk, the average zinc content in all studied varieties was 50.4 mg/kg, with 63.7 mg/ kg in the best variety 'OmGAU 100'. These values are higher than the target values of the international program Harvest Plus. 'Novosibirskaya 16' (49.4 mg/kg), 'Silach' (48.4 mg/kg), 'Line 4-10-16' (47.2 mg/ kg), 'Element 22' (46.3 mg/kg) and 'Lutescens 248/01' (46.0 mg/kg) were identif ied as being the best varieties. Signif icant possibilities for the production of wheat grain with high zinc content, which is in demand for the production of bread and pastry products with functional properties, were identif ied in the Western Siberian region.

6.
Vavilovskii Zhurnal Genet Selektsii ; 24(2): 131-138, 2020 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-33659792

RESUMEN

Stem rust in recent years has acquired an epiphytotic character, causing significant economic damage for wheat production in some parts of Western Siberia. On the basis of a race composition study of the stem rust populations collected in 2016-2017 in Omsk region and Altai Krai, 13 pathotypes in Omsk population and 10 in Altai population were identified. The race differentiation of stem rust using a tester set of 20 North American Sr genes differentiator lines was carried out. The genes of stem rust pathotypes of the Omsk population are avirulent only to the resistance gene Sr31, Altai isolates are avirulent not only to Sr31, but also to Sr24, and Sr30. A low frequency of virulence (10-25 %) of the Omsk population pathotypes was found for Sr11, Sr24, Sr30, and for Altai population - Sr7b, Sr9b, Sr11, SrTmp, which are ineffective in Omsk region. Field evaluations of resistance to stem rust were made in 2016-2018 in Omsk region in the varieties and spring wheat lines from three different sources. The first set included 58 lines and spring bread wheat varieties with identified Sr genes - the so-called trap nursery (ISRTN - International Stem Rust Trap Nursery). The second set included spring wheat lines from the Arsenal collection, that were previously selected according to a complex of economically valuable traits, with genes for resistance to stem rust, including genes introgressed into the common wheat genome from wild cereal species. The third set included spring bread wheat varieties created in the Omsk State Agrarian University within the framework of a shuttle breeding program, with a synthetic wheat with the Ae. tauschii genome in their pedigrees. It was established that the resistance genes Sr31, Sr40, Sr2 complex are effective against stem rust in the conditions of Western Siberia. The following sources with effective Sr genes were selected: (Benno)/6*LMPG-6 DK42, Seri 82, Cham 10, Bacanora (Sr31), RL 6087 Dyck (Sr40), Amigo (Sr24, 1RS-Am), Siouxland (Sr24, Sr31), Roughrider (Sr6, Sr36), Sisson (Sr6, Sr31, Sr36), and Fleming (Sr6, Sr24, Sr36, 1RS-Am), Pavon 76 (Sr2 complex) from the ISRTN nursery; No. 1 BC1F2 (96 × 113) × 145 × 113 (Sr2, Sr36, Sr44), No. 14а F3 (96 × 113) × 145 (Sr36, Sr44), No. 19 BC2F3 (96 × 113) × 113 (Sr2, Sr36, Sr44), and No. 20 F3 (96 × 113) × 145 (Sr2, Sr36, Sr40, Sr44) from the Arsenal collection; and the Omsk State Agrarian University varieties Element 22 (Sr31, Sr35), Lutescens 27-12, Lutescens 87-12 (Sr23, Sr36), Lutescens 70-13, and Lutescens 87-13 (Sr23, Sr31, Sr36). These sources are recommended for inclusion in the breeding process for developing stem rust resistant varieties in the region.

7.
Rev Sci Instrum ; 90(3): 033302, 2019 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-30927810

RESUMEN

The operation modes of a self-magnetically insulated ion diode were studied. The plasma formation at the anode surface was found to be accompanied by an electron leakage current due to the bipolar high voltage pulse. The emitting anode surface and ion beam density distribution were investigated at the ion diode output area. The characteristics of the electron leakage current are presented depending on the ion diode operation mode. The area of the emitting anode surface was approximately 25%-30% of the total anode area. The electron leakage current reaches up to 6.5 kA in the ion diode.

8.
Oncogene ; 26(43): 6261-8, 2007 Sep 20.
Artículo en Inglés | MEDLINE | ID: mdl-17452980

RESUMEN

Acetylation is thought to be a key event for p53 activation. We demonstrate that p14ARF-induced senescence of human mammary epithelial cells (MEC) is associated with p53 acetylation and requires hAda3, a component of histone acetyltransferase complexes and a p53 transcriptional coactivator. Expression of the N-terminal domain of hAda3 that binds p53 but not p300 blocked p14ARF-induced p53 acetylation and protected MECs from senescence. Consistent with these findings, the human papillomavirus 16 E6 mutant Y54D, which selectively targets hAda3 but not p53 for degradation and protects MECs from p14ARF-induced senescence, inhibited p53 acetylation. In H1299 cells, hAda3 overexpression increased p300-mediated p53 acetylation, which conversely decreased following small interfering RNA (siRNA) knockdown of hAda3. Moreover, depletion of hAda3 by siRNA inhibited endogenous p53 acetylation and accumulation of p21cip1 in response to ectopic p14ARF. These studies reveal that, in addition to its known ability to inhibit Mdm2-mediated p53 degradation, p14ARF signals through hAda3 to stimulate p53 acetylation and the induction of cell senescence.


Asunto(s)
Senescencia Celular/fisiología , Factores de Transcripción/metabolismo , Proteína p14ARF Supresora de Tumor/metabolismo , Proteína p53 Supresora de Tumor/metabolismo , Acetilación , Línea Celular Tumoral , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/metabolismo , Células Epiteliales/metabolismo , Humanos , Interferencia de ARN , Telomerasa/genética , Telomerasa/metabolismo , Factores de Transcripción/genética , Proteína p14ARF Supresora de Tumor/genética , Proteína p53 Supresora de Tumor/genética
9.
Exp Oncol ; 39(2): 155-156, 2017 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-29483495

RESUMEN

AIM: Classic activating mutations L858R and deletions in exon 19 (19del) in the gene for epidermal growth factor receptor (EGFR) are associated with sensitivity of the non-small cell lung cancer (NSCLC) to therapy with tyrosine kinase inhibitors (TKI). Insertions in EGFR exon 19 (19ins) are rare mutations in NSCLC; response of cases with 19ins to TKI is not well studied. Here we report a case of NSCLC with 19ins in a Russian patient who was treated with gefitinib. We also overview cases of 19ins reported in the literature. CASE DESCRIPTION: A 48 years old female Russian patient was diagnosed with adenocarcinoma of the lung (T3N2M1, stage IV). Mutation 19ins was detected in the tumor biopsy by fragment analysis and genotyped by Sanger sequencing as p.I744_K745insKIPVAI. Treatment with gefitinib (250 mg/day) resulted in clinical and radiological improvements scored as partial response that lasted 12 months. CONCLUSION: Treatment with gefitinib of lung adenocarcinoma that carries mutation EGFR 19ins can result in durable response.


Asunto(s)
Adenocarcinoma/tratamiento farmacológico , Adenocarcinoma/genética , Antineoplásicos/uso terapéutico , Receptores ErbB/genética , Exones , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/genética , Mutagénesis Insercional , Inhibidores de Proteínas Quinasas/uso terapéutico , Quinazolinas/uso terapéutico , Adenocarcinoma/diagnóstico , Adenocarcinoma del Pulmón , Antineoplásicos/administración & dosificación , Antineoplásicos/efectos adversos , Biopsia , Análisis Mutacional de ADN , Femenino , Gefitinib , Humanos , Neoplasias Pulmonares/diagnóstico , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Inhibidores de Proteínas Quinasas/administración & dosificación , Inhibidores de Proteínas Quinasas/efectos adversos , Quinazolinas/administración & dosificación , Quinazolinas/efectos adversos , Eliminación de Secuencia , Tomografía Computarizada por Rayos X
10.
J Natl Cancer Inst ; 88(12): 802-11, 1996 Jun 19.
Artículo en Inglés | MEDLINE | ID: mdl-8637046

RESUMEN

BACKGROUND: Nonmelanoma carcinomas of the skin represent the most frequent cancers among the Caucasian population worldwide. They occur with high frequency in renal allograft recipient patients after prolonged immunosuppression. PURPOSE: We analyzed tumors obtained from both immunosuppressed and nonimmunosuppressed patients for human papillomavirus (HPV) DNA. METHODS: Twenty-nine specimens of nonmelanoma carcinomas of the skin were obtained from 19 renal allograft recipient patients; these included 20 specimens of squamous cell carcinoma (SCC) from 11 patients, five specimens of basal cell carcinoma (BCC) from four patients, and four specimens of carcinoma in situ (CIS) from four patients. Forty-one specimens of nonmelanoma carcinomas of the skin were obtained from 32 nonimmunosuppressed patients; these included 26 SCC specimens from 19 patients, 11 BCC specimens from nine patients, and four keratoacanthoma (benign epithelial tumor) specimens from four patients. A polymerase chain reaction method involving use of degenerate oligonucleotide primers, in which the conserved region of the open reading frame of the HPV L1 (major capsid protein) gene is amplified, was used to amplify total cellular DNA purified from individual tumors. The DNA of each specimen was subjected to 16 different amplification reactions; different primer combinations were used in order to increase the sensitivity and specificity of HPV detection. Resulting products were probed with a radioactively labeled, degenerate oligonucleotide. HPV-specific DNA was either sequenced directly after elution from the gel or amplified with semi-nested, degenerate primers, after which the products were cloned and sequenced. Sequences were compared with all known papillomavirus sequences. RESULTS: Thirteen (65%) of the 20 SCC specimens and three of the five BCC specimens from immunosuppressed (renal allograft recipient) patients contained identifiable HPV-related sequences, among them 13 putative novel HPV genomes. In addition, all other malignant tumor specimens from this patient group revealed faint signals upon amplification and hybridization; the origin of these signals has not been identified in the present study. In nonimmunosuppressed patients, eight (31%) of 26 SCC specimens and four (36%) of 11 BCC specimens contained sequences of HPV types. Two putative novel HPV sequences could be identified in this group. Faint signals of yet undetermined origin were observed in eight of the SCC specimens and in two of the BCC specimens. Two of four keratoacanthoma specimens contained sequences of known HPV type. (Keratoacanthoma is a nonmalignant lesion for which the natural history has not been defined.) The spectrum of HPV types in both groups of patients differed substantially. CONCLUSIONS: These data point to the frequent presence of HPV sequences in SCCs and BCCs of the skin. The etiologic relationship of these infections to the respective malignant tumors remains to be evaluated. IMPLICATIONS: The presence of HPV DNA in a large percentage of specimens of nonmelanoma carcinomas of the skin from immunosuppressed patients, as well as from nonimmmunosuppressed patients, renders a papillomavirus infection as a possible factor in the etiology of this disease.


Asunto(s)
ADN Viral/análisis , Trasplante de Riñón , Papillomaviridae/aislamiento & purificación , Infecciones por Papillomavirus/complicaciones , Neoplasias Cutáneas/virología , Infecciones Tumorales por Virus/complicaciones , Verrugas/complicaciones , Secuencia de Aminoácidos , Carcinoma in Situ/virología , Carcinoma Basocelular/virología , Carcinoma de Células Escamosas/virología , Humanos , Huésped Inmunocomprometido , Queratoacantoma/virología , Datos de Secuencia Molecular , Papillomaviridae/genética , Reacción en Cadena de la Polimerasa/métodos , Sensibilidad y Especificidad , Verrugas/virología
11.
Cancer Res ; 54(17): 4610-3, 1994 Sep 01.
Artículo en Inglés | MEDLINE | ID: mdl-8062252

RESUMEN

A total of 118 biopsies from skin lesions of 46 renal allograft patients was analyzed for human papillomavirus (HPV) DNA by polymerase chain reaction with degenerate primers and also partially by subsequent sequencing of the amplified fragment. Sixty-two % of the benign proliferations (31 of 50) contained DNA of known HPV types as well as HPV sequences related to a number of epidermodysplasia verruciformis-associated HPV types. HPV DNA sequences were found in 14 (56%) of 25 biopsies from squamous cell and basal cell carcinomas. One squamous cell carcinoma contained HPV 41 DNA. A novel 640-base pair fragment sharing homology with HPV 29 (82.7%) was found in 15% (3 of 20) of squamous cell carcinomas, in 9.4% (3 of 32) of dysplastic warts and in 8.5% (4 of 47) common warts. The remaining positive carcinoma biopsies contained HPV-related DNA in such a low copy number that additional analysis is required. The identification of new HPV types in skin cancers of immunosuppressed patients (other than epidermodysplasia verruciformis patients) further expands the spectrum of HPV-linked human malignancies and permits new approaches to study the pathogenesis of skin cancers.


Asunto(s)
Carcinoma Basocelular/microbiología , Carcinoma de Células Escamosas/virología , ADN Viral/aislamiento & purificación , Huésped Inmunocomprometido , Trasplante de Riñón , Papillomaviridae/aislamiento & purificación , Neoplasias Cutáneas/virología , Piel/virología , Verrugas/virología , Secuencia de Aminoácidos , Secuencia de Bases , Sondas de ADN de HPV , Humanos , Datos de Secuencia Molecular
12.
Acta Virol ; 35(1): 71-80, 1991 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-1683119

RESUMEN

Recombinant plasmid DNA was used as a probe to detect tick-borne encephalitis (TBE) virus RNA during incubation period, acute disease and persistent infection of syrian hamsters. Within the first three weeks post-infection the results of direct virus isolation and RNA detection in the brain agreed by a rate of 100%, the virus titre ranging between 10(1.9) to 10(10.5) LD50/ml and viral RNA concentration at 1-1000 pg. At the same time TBE virus RNA was detected in the spleen when the virus titre was greater than or equal to 10(6.5) LD50/ml. By 8 months post infection (p.i.) viral RNA was found in the brain, liver, and spleen in the absence of infectious TBE virus. No viral RNA was present in the thymus. In addition, electron microscopic findings in hamster brain confirmed the hypothesis that TBE virus persistence was accompanied by formation of virus-specific structures but impaired virion maturation.


Asunto(s)
Sondas de ADN , ADN Viral , Virus de la Encefalitis Transmitidos por Garrapatas/aislamiento & purificación , Encefalitis Transmitida por Garrapatas/microbiología , ARN Viral/análisis , Enfermedad Aguda , Animales , Encéfalo/microbiología , Encéfalo/patología , Enfermedad Crónica , Cricetinae , ADN/genética , ADN Recombinante/genética , Virus de la Encefalitis Transmitidos por Garrapatas/genética , Encefalitis Transmitida por Garrapatas/patología , Hígado/microbiología , Mesocricetus/microbiología , Hibridación de Ácido Nucleico , Bazo/microbiología , Timo/microbiología
13.
Bioorg Khim ; 20(11): 1196-205, 1994 Nov.
Artículo en Ruso | MEDLINE | ID: mdl-7880179

RESUMEN

The non-radioactive reverse dot-blot method was used for the detection of tick-borne encephalitis virus (TBEV) in clinical specimens. The method involves reverse transcription (RT) and polymerase chain reaction (PCR) using a pair of biotin-labelled oligonucleotide primers. These primers flank a region in the gene of the envelope protein E, which is more conserved than other regions, and initiate the polymerisation with RNAs of all the investigated strains. The amplified cDNA was captured from solution on a solid support using complementary oligonucleotides covalently bound to a polyamide membrane. The biotin labels of the resulting hybrids were visualized by means of the streptavidin-horseradish peroxidase conjugate. The detection limit of the test was about 10(3)-10(4) molecules of target RNA. The sensitivity was comparable to that obtained by dot-hybridization of PCR-product with 32P-labelled DNA probe. The method was used for the detection of RNA in specimens of tick and blood.


Asunto(s)
Virus de la Encefalitis Transmitidos por Garrapatas/aislamiento & purificación , Secuencia de Bases , Cartilla de ADN , Sondas de ADN , ADN Complementario , Virus de la Encefalitis Transmitidos por Garrapatas/genética , Genoma Viral , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , ARN Viral/análisis
14.
Vopr Virusol ; 35(6): 474-8, 1990.
Artículo en Ruso | MEDLINE | ID: mdl-2082550

RESUMEN

Synthetic deoxyoligonucleotides complementary to different regions of genome RNA of tick-borne encephalitis (TBE) Sofjin virus strain were used to differentiate TBE virus strains. Nine TBE strains isolated in different geographical areas from different sources and several viruses of the TBE subgroup were tested. The probes revealed genetic heterogeneity of TBE strains. The probes complementary to different regions of the genome had different specificity. The pattern of hybridization of TBE virus strains with a panel of 11 oligonucleotide probes correlated significantly with the source of the virus strain and to a smaller extent with the geographical isolation site.


Asunto(s)
Virus de la Encefalitis Transmitidos por Garrapatas/aislamiento & purificación , Hibridación de Ácido Nucleico , Sondas de Oligonucleótidos , ARN Viral/genética , Animales , Virus de la Encefalitis Transmitidos por Garrapatas/genética , Encefalitis Transmitida por Garrapatas/microbiología , Filtración/instrumentación , Ratones , Sondas de Oligonucleótidos/síntesis química , Sondas de Oligonucleótidos/aislamiento & purificación
15.
Vopr Virusol ; 37(1): 53-6, 1992.
Artículo en Ruso | MEDLINE | ID: mdl-1384240

RESUMEN

Experiments on molecular hybridization were carried out using a panel of 11 deoxyoligonucleotide probes complementary to different parts of tick-borne encephalitis (TBE) virus, strain Sophyin, genome. Under study were the TBE virus strains differing by 3 criteria: (1) source of isolation (patients with acute and chronic TBE, Ixodes persulcatus and D. nuttalli ticks, small mammals); (2) serotype (eastern and Siberian Aina/1448), (3) virulence for Syrian hamsters. RNA of all the strains was hybridized with kDNA, 90% of strains with probe Sh5 complementary to protein E gene, nucleotide positions 1285-1311. The highest differentiating capacity was observed with probes P131 and Sh3 complementary to genes of proteins ns2b and M. These probes reacted with RNA of 100% of highly virulent strains of the eastern serotype and only with 20-30% of strains of the Aina/1448 serotype of lower virulence. A certain differentiating capacity was demonstrated by probes Sh2 and P10 complementary to genes of prm and C proteins: they hybridized with RNA of 80% of eastern serotype strains highly virulent for hamsters and with only 20% of Aina/1448 serotype strains of low virulence. The panel of probes used revealed no significant differences among strains in relation to their isolation source, with the exception of a strain isolated from D. nuttalli ticks which reacted only with kDNA and probe P2 complementary to nsI protein gene, but not with other probes. The TBE virus strains isolated from patients with chronic TBE were shown to represent a genetically heterogeneous group.


Asunto(s)
Antígenos Virales/análisis , Virus de la Encefalitis Transmitidos por Garrapatas/inmunología , Variación Genética/inmunología , Sondas de Oligonucleótidos , Animales , Antígenos Virales/genética , Encéfalo/microbiología , Cricetinae , Virus de la Encefalitis Transmitidos por Garrapatas/clasificación , Virus de la Encefalitis Transmitidos por Garrapatas/genética , Virus de la Encefalitis Transmitidos por Garrapatas/patogenicidad , Encefalitis Transmitida por Garrapatas/microbiología , Variación Genética/genética , Genoma Viral , Mesocricetus , Ratones , Hibridación de Ácido Nucleico , Fenotipo , ARN/genética , ARN/aislamiento & purificación , ARN Viral/genética , Pase Seriado , Serotipificación , Garrapatas/microbiología , Virulencia/genética , Virulencia/inmunología
16.
Vopr Virusol ; 36(1): 27-31, 1991.
Artículo en Ruso | MEDLINE | ID: mdl-1713371

RESUMEN

Nucleic acid spot hybridization with cloned cDNA of tick-borne encephalitis (TBE) virus, strain Sofjin, was used to differentiate strains of TBE and other flaviviruses. The cDNA probe reacted with strains of TBE and flaviviruses of TBE subgroup with the exception of Powassan virus. The probe did not react with viruses of Japanese encephalitis and Gendue subgroups. The viruses of TBE subgroup and some strains of TBE virus were differentiated from TBE strain Sofjin by thermal stability of RNA-DNA hybrids. Negishi and Louping ill viruses were found to be most closely related to TBE strain Sofjin among viruses of the TBE subgroup.


Asunto(s)
ADN Viral/genética , Virus de la Encefalitis Transmitidos por Garrapatas/clasificación , Hibridación de Ácido Nucleico/genética , ARN Viral/genética , Animales , Encéfalo , ADN/genética , Sondas de ADN/aislamiento & purificación , Virus de la Encefalitis Transmitidos por Garrapatas/genética , Genes Virales/genética , Técnicas Genéticas , Ratones , ARN/aislamiento & purificación
17.
Vopr Virusol ; 37(5-6): 248-52, 1992.
Artículo en Ruso | MEDLINE | ID: mdl-1290224

RESUMEN

Hybridization experiments with RNA of 143 tick-borne encephalitis (TBE) virus strains isolated in different parts of the distribution area were used to study the reactivity of kDNA- and a set of 10 synthetic deoxyoligonucleotide probes. The kDNA probe under certain conditions was shown to hybridize with RNA of all the strains under study, and under other (strict) hybridization conditions did so selectively with a small number of strains. The capacity of oligonucleotide probes for hybridization with RNA of TBE virus strains varied from 12% to 100%. The differences in the hybridization activity of kDNA- and oligonucleotide probes complementary to the genomes of the Sophyin strain (Far-Eastern subtype) and Neudorffle strain (Western subtype) with TBE virus strains were used for differentiation of the strains into six genetic variants. Comparison of the reactivity of molecular probes in experiments with RNA of TBE virus strains and viruses of the TBE complex showed that the differences of the strains belonging to different genetic variants from the prototype Sophyin strain were comparable to those of some members of the TBE complex, with the exception of Powassan virus. These data attest to the necessity of further studies dealing with specification of the taxonomy of TBE complex viruses.


Asunto(s)
Sondas de ADN/genética , ADN Viral/genética , ADN/genética , Virus de la Encefalitis Transmitidos por Garrapatas/genética , Genoma Viral , Sondas de Oligonucleótidos/genética , ARN Viral/genética , Animales , Secuencia de Bases , Virus de la Encefalitis Transmitidos por Garrapatas/aislamiento & purificación , Variación Genética/genética , Datos de Secuencia Molecular , Hibridación de Ácido Nucleico/métodos , ARN Viral/aislamiento & purificación , Federación de Rusia , Garrapatas/microbiología
18.
Vopr Virusol ; 37(5-6): 252-6, 1992.
Artículo en Ruso | MEDLINE | ID: mdl-1290225

RESUMEN

Geographic distribution of 185 tick-borne encephalitis (TBE) virus strains isolated in 8 physico-geographic areas and classified into six genetic variants was analysed. The strains of genetic variant I homologous to the Sophyin prototype strain were found to occur predominantly in the Far East and also frequently found in Western and North-Western parts of the East European plain. The vast territories from lake Baikal in the East to Ukraine in the West harbor mostly the strains significantly different from the Far-Eastern Sophyin strain. Hybridization experiments with oligonucleotide probes specific for the Neudorffle strain showed that the strains genetically similar to the virus of central European encephalitis occurred also in Eastern Europe and Western Siberia. It is concluded that a relationship exists between genetic types of TBE virus and their geographic origin.


Asunto(s)
Virus de la Encefalitis Transmitidos por Garrapatas/genética , Variación Genética/genética , Animales , Animales Recién Nacidos , ADN/genética , Sondas de ADN/genética , ADN Viral/genética , Reservorios de Enfermedades , Virus de la Encefalitis Transmitidos por Garrapatas/aislamiento & purificación , Genoma Viral , Ratones , Sondas de Oligonucleótidos/genética , ARN Viral/genética , Federación de Rusia , Garrapatas/microbiología
19.
Vopr Virusol ; 35(1): 57-9, 1990.
Artículo en Ruso | MEDLINE | ID: mdl-2194370

RESUMEN

Comparative studies of the diagnostic value of three express methods for detection of tick-borne encephalitis virus in ticks (fluorescent antibody technique (FAT) enzyme immunoassay (EIA), and molecular hybridization of nucleic acids) and the traditional method (bioassay in white mice) showed all the three express methods to be rapid, specific, sensitive, and useful for large-scale epidemiological surveys. Notable was the high effectiveness of the method of nucleic acids hybridization which was not inferior to bioassays in suckling mice and exceeded FAT and EIA. The results of the latter seem to be affected by antigenic variations among tick-borne encephalitis virus strains.


Asunto(s)
Virus de la Encefalitis Transmitidos por Garrapatas/aislamiento & purificación , Animales , Técnica del Anticuerpo Fluorescente , Técnicas para Inmunoenzimas , Ratones , Hibridación de Ácido Nucleico
20.
Artículo en Ruso | MEDLINE | ID: mdl-11210648

RESUMEN

The comparative evaluation of the PCR test "Polimik" (Research and Production Firm "Litekh", Moscow) and the PCR test of the Novosibirsk Institute of Bioorganic Chemistry (NIBC) was carried out. The results obtained with the use of the PCR test "Polimik" and the PCR test of the NIBC of the detection of C. trachomatis and M. hominis coincided in 97.8% and 97.4% of cases. For U. urealyticum, the coincidence of the results of both PCR tests was 81.2%. Among women who visited gynecologists for reproductive function disturbances, the use of the PCR tests made it possible to detect C. trachomatis in 19 (5.5%) out of 343 cases, U. urealyticum in 96 (39.0%) out of 246 cases and M. hominis in 25 (16.9%) out of 148 cases. The results of the investigation revealed that the occurrence of C. trachomatis infection in Novosibirsk was comparable with that in other regions of the world among the low-risk groups of the population. The detection frequency of M. hominis and U. urealyticum with the use of the PCR tests showed that the occurrence of infections caused by these causative agents coincided with the data obtained in other countries.


Asunto(s)
Chlamydia trachomatis/genética , ADN Bacteriano/análisis , Infertilidad Femenina/complicaciones , Mycoplasma hominis/genética , Enfermedades Bacterianas de Transmisión Sexual/microbiología , Ureaplasma urealyticum/genética , Femenino , Humanos , Reacción en Cadena de la Polimerasa/métodos , Enfermedades Bacterianas de Transmisión Sexual/complicaciones
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