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Query: UNIPROT:P06889 (
Mol
)
630,302
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
The biotin-labeled DNA probes were constructed on the basis of the hybrid bacteriophage M13nip 9 single-stranded DNA containing the fragments of the
hepatitis A
viral cDNA. The probes were biotin treated by chemical modification of the DNA by the peraminating reagent or photochemically. The labeled DNA probes were used in molecular hybridization experiments with the nuclear acids fixed on the nitrocellulose filters. The biotin treated DNA was determined by the avidin-gold colloid conjugate with the subsequent physical silver amplification or by the streptavidin-alkaline phosphatase conjugate. The sensitivity of both probes was identical and permitted the determination of 5 x 10(-11)-5 x 10(-12) g of the control DNA and 10(-9) g of the
hepatitis A
virus. The developed test systems were used for detection of the viral RNA in blood from patients.
Mol
Gen Mikrobiol Virusol
PMID:[Use of non-radioactive biotin-labelled probes for detecting hepatitis A virus]. 133 51
The synthetic oligonucleotide sequence 3'CTCCTCGTACTTTA-5' complementing
hepatitis A
virus RNA was compared with cDNA probes in identification of viral genomic RNA. The clinical materials from patients in the 1-2 weeks of jaundice were screened. High specificity of the technique was demonstrated. Possibility of clinical using of the blot hybridization technique is discussed.
Mol
Gen Mikrobiol Virusol 1987 Sep
PMID:[Detection of genomic RNA of the hepatitis A virus in clinical samples using the blot hybridization method]. 282 40
Three fragments of a DNA copy complementary to tre
hepatitis A
virus RNA have been isolated from the pHAV-VPI-22 plasmid and cloned in M 13 mp 8 vector. The 140, 250 and 390 b. p. fragments corresponded to the viral genome fragment coding for VP1 protein. Single-stranded DNAs of the hybrid phages with 250 b. p. and 390 b. p. inserts have been used for radioactive probe synthesis. The specificity of the probes for viral RNA was confirmed in hybridization with
hepatitis A
virus RNA purified from the virus-infected cell culture. The hybridization was shown to detect up to 10(-12)-10(-13) g of the virus. A principal possibility to use the probes in diagnostic purposes was demonstrated by the virus detection in blood samples obtained from patients with
hepatitis A
infection.
Mol
Gen Mikrobiol Virusol 1988 Jul
PMID:[Construction of probes for hybridization with hepatitis A virus RNA based on phage M13]. 284 93
The propagation time-course of
hepatitis A
virus (HAV, strain HAS-15) in continuous culture of the foetal rhesus monkey kidney cells (FRhK-4) was investigated. The HAV infectivity and viral RNA content in the infected cells reached the maximal level 5-8 days after infection, while accumulation of
hepatitis A
antigen (HAAg) continued for 2-3 weeks more. Viral particles with the densities 1.27-1.28 g/cm3 and 1.18-1.22 g/cm3 were isolated from the infected cells as well as the mature virions with the buoyant density 1.33-1.34 g/cm3 in CsCl. The concurrent accumulation of mature virus and "light" particles (1.18-1.22 g/cm3) was registered during infection. Viral particles with the density 1.27-1.28 g/cm3 accumulated predominantly from the 14th to the 21st-24th days after infection. The mature virions (1.34 g/cm3) as well as the particles with the density 1.24-1.25 g/cm3 were isolated from supernatant precipitated by ammonium sulphate. The HAAg activity of both fractions increased progressively in equal proportion in course of infection.
Mol
Gen Mikrobiol Virusol 1987 Jan
PMID:[Dynamics of the accumulation of virus particles with different physico-chemical properties in FRhK-4 cells infected with hepatitis A virus]. 303 92
Four types of virus-specific particles with different sedimentation coefficients and buoyant densities in CsCl were shown to be accumulated in
hepatitis A
virus (strain HAS-15) infected fetal rhesus monkey kidney cells (FRhK-4 line). Unlike the mature virions (155S, 1.34 g/cm3), cell-associated isosedimenting 92 S-particles (buoyant densities of 1.30 and 1.20 g/cm3) proved to be sensitive to lipase action. Particles of all four types were shown to contain similar sets of polypeptides, and, with the exception of "empty" 1.30 g/cm3-particles, appeared to be "full" under the immune electron microscopic examination. The viral RNA was unequivocally identified by the molecular hydridization test only in the mature virions.
Mol
Gen Mikrobiol Virusol 1987 Mar
PMID:[Properties of hepatitis A virus particles produced during infection in vitro]. 303 82
The results of adaptation of
hepatitis A
viral strain JaM-55 to the culture of embryo kidney cells FRhk-4 from macaque Rhesus are presented. The viral strain was isolated from a M. fascicularis suffering from spontaneous hepatitis. Before inoculating the cell culture the virus was passaged twice in the M. arctoides capable of reproducing hepatitis. FRhk-4 cell line inoculation by the monkey liver extract, containing the strain HAV-YaM-55, resulted in isolation of single viral particles of
hepatitis A
in the preparations obtained at the first 3 passages by the 28-31 day of cultivation. Beginning from the fourth passage the abrupt increase in the number of viral particles and
hepatitis A
antigen was registered. There were no traces of cytopathogenic effect at any level of viral passages in the inoculated cell culture. The adapted virus contains
hepatitis A
viral RNA identified by spot hybridization with the cloned cDNA of
hepatitis A
virus.
Mol
Gen Mikrobiol Virusol 1987 May
PMID:[Adaptation of hepatitis A virus strain (JaM-55) originally pathogenic for monkeys to a cell culture]. 303 62
The purpose of this study was to determine the efficiency of semi-nested PCR in detecting
hepatitis A
virus (HAV) RNA. During a 2-year period (1990-1991), HAV RNA was searched for in shellfish from the French Brittany coasts using cRNA and vRNA probes. In January 1992, at the time of a
hepatitis A
outbreak, 28 stool samples were collected from infected patients (18 adults, 10 children) with anti-HAV IgM. Four samples from subjects with negative HAV serology were used as negative controls. Nucleic acid amplification (reverse-transcription-semi-nested PCR) was performed to detect HAV in stool. HAV RNA was purified by phenol-chloroform extraction and converted to cDNA using reverse transcriptase (Mu-MLV). After amplification, PCR products were visualized on an ethidium-bromide-stained gel and confirmed by hybridization with a specific digoxigenin-labelled oligoprobe. Samples were also studied by molecular hybridization with cRNA and vRNA probes. After onset of the illness, HAV RNA was detected over a longer time period by semi-nested PCR (16/28) than by hybridization (0/28). Even though biological diagnosis of
hepatitis A
will continue to rely on the detection of anti-HAV IgM, PCR should be useful in certain clinical cases (diagnosis of relapse) and for epidemiological and environmental monitoring of viruses.
Mol
Cell Probes 1994 Apr
PMID:Development of RT-semi-nested PCR for detection of hepatitis A virus in stool in epidemic conditions. 793 9
Several isoforms of the wild-type and three mutant
hepatitis A
virus (HAV) 3C proteinases have been isolated and characterized. The active site cysteine residue (residue 172) was found to be responsible for the formation of some of these isoforms. The double mutant C24S/C172A of the HAV 3C proteinase, in which both cysteine residues have been replaced by site-directed mutagenesis, was crystallized. The crystals belong to the hexagonal space group P6(1)22 (or its enantiomorph, P6(5)22) with unit cell dimensions a = b = 65.2 A, c = 246.1 A and diffract X-rays to 2.3 A resolution.
J
Mol
Biol 1993 Dec 05
PMID:Hepatitis A virus 3C proteinase: some properties, crystallization and preliminary crystallographic characterization. 825 82
Cytochrome P450 (CYP) 2A5 is involved in the metabolism of carcinogens like aflatoxin B1 and N-nitrosodiethylamine (NDEA), and CYP2A5 levels are increased in some pathological states of the liver (e.g.,
infectious hepatitis
and porphyria). We analyzed the expression of CYP2A5 during experimental liver carcinogenesis in three different mouse strains (C3H/He, C57BL/6J, and B6C3F1) with immunohistochemical techniques and in situ hybridization. In normal liver, CYP2A5 protein and mRNA were detected in centrilobular hepatocytes only. Phenobarbital treatment increased the number of CYP2A5-positive centrilobular hepatocytes and the CYP2A5-positive areas were extended into the middle zone in all strains, but periportal hepatocytes remained negative. Fifty percent of the spontaneous foci in untreated mice, over 90% of the foci in mice treated with NDEA or phenobarbital and all of the hepatocellular adenomas and carcinomas displayed positive immunostaining and a strong CYP2A5 mRNA signal by in situ hybridization. In the liver tumors metastasized to the lung, expression of CYP2A5 had largely disappeared. CYP2A5 expression in neoplastic and putative preneoplastic lesions, although sometimes heterogeneous, was apparently independent of the typical zonal expression pattern in normal tissue. As expected, the C57BL/6J mice developed fewer foci and tumors than the C3H/He and B6C3F1 mice, but the phenotype of CYP2A5 overexpression was similar in all the strains. Our data suggest that the increased expression of CYP2A5 may play an important role in the development of liver cancer in mice and may be used as a novel marker for spontaneous and NDEA-induced mouse liver foci.
Mol
Carcinog 1998 Aug
PMID:Expression of cytochrome P450 2A5 in preneoplastic and neoplastic mouse liver lesions. 972 15
The purpose of this study was to determine the frequency of infection by
hepatitis A
, B, C, D, E, and G in liver biopsy specimens from symptomatic patients and to correlate viral localization with the expression of interferon tau, interleukin 4, and tumor necrosis factor messenger RNA. Tissue biopsy specimens were taken from 78 patients as follows: 14 patients with transplants, 23 patients with cirrhotic livers, and 41 patients with chronic hepatitis. At least one of the hepatitis viruses was detected in 60 of 78 (77%) specimens; multiple infection was evident in 18 of 78 (23%) specimens. The overall incidence of the different viruses was as follows: 8%
hepatitis A
, 3% hepatitis B, 52% hepatitis C, 1% hepatitis D, 24% hepatitis E, 18% hepatitis G. Throughout each category, hepatitis C was the most common virus detected. No histologic variable correlated with either the percentage of infected hepatocytes per lobule or nodule or with the specific viral type. The cytokines localized to monocytes or lymphocytes adjacent to infected hepatocytes. These results demonstrate that viral infection is present in most biopsy specimens of patients with chronic hepatitis and liver transplants and that hepatitis C, E, and G account for most of the infections. The results also suggest that direct viral infection in conjunction with expression of different cytokines is important in the pathophysiology of viral-induced liver disease.
Diagn
Mol
Pathol 1998 Oct
PMID:Histologic distribution of hepatitis A, B, C, D, E, and G with concomitant cytokine response in liver tissue. 999 Apr 85
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