RESUMO
Immunogold labeling was used to localize the core protein of small dermatan sulfate proteoglycan (DS-PG) on the surface of cultured human fibroblasts. At 4 degrees C, DS-PG core protein was uniformly distributed over the cell surface. At 37 degrees C, gold particles either became rearranged in form of clusters or remained associated with fibrils. Double-label immunocytochemistry indicated the co-distribution of DS-PG core protein and fibronectin in the fibrils. In an enzyme-linked immunosorbent assay, binding of DS-PG from fibroblast secretions and of its core protein to fibronectin occurred at pH 7.4 and at physiological ionic strength. Larger amounts of core protein than of intact proteoglycan could be bound. Fibronectin peptides containing either the heparin-binding domain near the COOH-terminal end or the heparin-binding NH2 terminus were the only fragments interacting with DS-PG and core protein. Competition and replacement experiments with heparin and dermatan sulfate suggested the existence of adjacent binding sites for heparin and DS-PG core protein. It is hypothesized that heparan sulfate proteoglycans and DS-PG may competitively interact with fibronectin.
Assuntos
Membrana Celular/metabolismo , Proteoglicanas de Sulfatos de Condroitina/metabolismo , Condroitina/análogos & derivados , Dermatan Sulfato/metabolismo , Fibronectinas/metabolismo , Proteoglicanas/metabolismo , Membrana Celular/análise , Membrana Celular/ultraestrutura , Proteoglicanas de Sulfatos de Condroitina/análise , Dermatan Sulfato/análise , Dermatan Sulfato/farmacologia , Fibroblastos , Fibronectinas/análise , Heparina/metabolismo , Heparina/farmacologia , Humanos , Concentração de Íons de Hidrogênio , Microscopia Eletrônica , TemperaturaRESUMO
The newly recognized human endogenous vasoconstrictive dinucleotides, diadenosine pentaphosphate (AP5A) and diadenosine hexaphosphate (AP6A), were tested for growth stimulatory effects in rat mesangial cells (MC). Both AP5A and AP6A stimulated growth in micromolar concentrations. The growth stimulatory effect exceeded that of ATP, alpha,beta-methylene ATP, adenosine 5'-O-(3-thio)triphosphate and UTP. Both diadenosine phosphates potentiated the growth response to platelet-derived growth factor, but not to insulin-like growth factor-1. To further elucidate the site of action in the cell cycle, RNA and protein synthesis were assessed. AP5 and AP6A stimulated protein synthesis, but not RNA formation. Furthermore, both agents increased cytosolic free Ca2+ concentration. It is concluded that AP5A and AP6A may play a regulatory role in MC growth as progression factors and possibly modify MC proliferation in glomerular disease.
Assuntos
Fosfatos de Dinucleosídeos/farmacologia , Mesângio Glomerular/citologia , Adenosina , Trifosfato de Adenosina/farmacologia , Animais , Cálcio/metabolismo , Divisão Celular/efeitos dos fármacos , Células Cultivadas , DNA/biossíntese , Sinergismo Farmacológico , Técnicas In Vitro , Masculino , Fator de Crescimento Derivado de Plaquetas/farmacologia , Ratos , Ratos Endogâmicos WKY , Uridina Trifosfato/farmacologiaRESUMO
Epidemiological, pathological, serological and virological investigations are reported on turkey haemorrhagic enteritis virus (THEV) infection in Hungarian turkey flocks. The pathogenesis of infection in experimentally infected turkeys and chickens, as well as the usefulness of polymerase chain reaction (PCR)/sequencing method for epidemiological investigation and for the differentiation of vaccine and field strains of THEV was also studied. Since the first recognition of the disease in Hungary in the late 1970s, until recently the disease has been diagnosed sporadically in its mild form. In the last few years (2000-2005), however, the number of outbreaks and the severity of the disease increased (9-23 affected flocks/year). Most of the outbreaks occurred at the age of 6 to 8 weeks and was complicated with Escherichia coli infection. The antibody levels to THEV in turkey flocks gradually declined till 5-7 weeks of age, and then they increased sharply due to natural infection with THEV. The immune response to vaccination (at 5 weeks of age) showed no significant antibody level increase one week postvaccination, but four weeks later the antibody level reached high values and then remained at this high level. The agar gel immunodiffusion (AGID) test to detect turkey adenovirus A (TAdV-A) antigen and PCR methods for THEV-specific DNA gave similarly positive results if spleens with pathognomonic lesions were tested; however, PCR proved to be more sensitive in cases with less characteristic pathological lesions. Nucleotide sequence alignment of PCR products amplified from Hungarian field strains and the Domermuth vaccine strain and that of the published THEV hexon sequences in GenBank database revealed slight differences between the sequences.
Assuntos
Infecções por Adenoviridae/veterinária , Surtos de Doenças/veterinária , Doenças das Aves Domésticas/epidemiologia , Doenças das Aves Domésticas/virologia , Siadenovirus/isolamento & purificação , Perus , Infecções por Adenoviridae/epidemiologia , Infecções por Adenoviridae/virologia , Animais , Antígenos Virais/sangue , DNA Viral/química , DNA Viral/genética , Hungria/epidemiologia , Reação em Cadeia da Polimerase/veterinária , Siadenovirus/genéticaRESUMO
New advances in the field of genetic characterization of adenoviruses originating from different animal species are summarized. Variations seen in the host range and specificity, pathogenicity, genomic arrangement or gene complement are much wider than expected based on previous studies of human adenoviruses. Several exceptional adenoviruses from the two traditional conventional genera are now removed, and proposed to form at least two new genera. The eventual host origin of the new genera, however, is not clarified. Novel results from the genomic and phylogenetic analyses of adenoviruses originating from lower vertebrate species (including reptiles, amphibians and fish) seem to imply that probably five major clusters of adenoviruses exist corresponding to the five major classes of Vertebrata. Adenoviruses, which are now suspected to have common origin with enterobacterium phages from the family Tectiviridae, are perhaps very ancient indeed, and may have undergone a co-evolution with vertebrate hosts.
Assuntos
Adenoviridae/genética , Evolução Molecular , Adenoviridae/patogenicidade , Animais , Genoma Viral , Interações Hospedeiro-Parasita , HumanosRESUMO
Until now, the only published ovine adenovirus DNA sequence was the complete genome of ovine adenovirus isolate 287 (OAV287) which, compared to other mammalian adenoviruses, possesses strikingly unique genomic organisation and should properly be classified into a new adenovirus genus. The protease gene sequence of ovine adenovirus type 3 (OAdV-3) was determined and analysed. The results of phylogenetic analysis of the 205 residue long protein demonstrated that OAdV-3 belongs to the genus Mastadenovirus, and is surprisingly closely related to bovine adenovirus type 2. In spite of the common host origin, the evolutionary distance between OAdV-3 and OAV287 proved to be great suggesting that sheep, similarly to cattle and fowl, might be infected by distantly related adenoviruses belonging to different genera.
Assuntos
Endopeptidases/genética , Mastadenovirus/classificação , Mastadenovirus/enzimologia , Filogenia , Doenças dos Ovinos/virologia , Infecções por Adenoviridae/veterinária , Sequência de Aminoácidos , Animais , Sequência de Bases , Bovinos , Endopeptidases/química , Endopeptidases/metabolismo , Humanos , Mastadenovirus/genética , Dados de Sequência Molecular , Análise de Sequência de DNA , OvinosRESUMO
The complete genome of a bovine adenovirus (BAV) type 10 isolate was molecularly cloned and partially sequenced. The encoded proteins were predicted by computer analysis of the DNA sequences of the ends or the entire length of the cloned viral fragments, and thus a rough genetic map was constructed. The protease gene of BAV-10 was completely sequenced and used in phylogenetic analysis. Based on the results of the phylogenetic analysis, and the location and presence of certain genes thought to be specifically characteristic of subgroup 1 or subgroup 2 BAVs, it could be concluded that, in spite of the striking similarity in certain biological properties, BAV-10 is not related to subgroup 2 BAVs as originally described. It does not however fit clearly into subgroup 1 either, the members of which show closer relationship with human adenoviruses. BAV-10 therefore should best be considered as the first member of a third subgroup of BAVs.
Assuntos
Endopeptidases/genética , Mastadenovirus/classificação , Mastadenovirus/genética , Filogenia , Adenovirus Caninos/genética , Adenovírus Humanos/genética , Animais , Aviadenovirus/genética , Sequência de Bases , Bovinos , Clonagem Molecular , Endopeptidases/química , Humanos , Mastadenovirus/enzimologia , Dados de Sequência Molecular , Mapeamento por Restrição , Análise de Sequência de DNA , SorotipagemRESUMO
The DNA sequence of the genome of bovine adenovirus type 2 (BAdV-2) was determined between map units 42.5 and 50. By sequence analysis and homology search, the genes of five structural proteins were identified within this region: the penton base protein (III; partial sequence), the major core protein precursor (pVII), the minor core protein (V), the mu core protein precursor (pX) and the hexon associated protein precursor (pVI; partial sequence). The putative polypeptides were compared to their known counterparts from other adenoviruses. The existence of protein V and the presence and structure of certain protease cleavage recognition sites confirmed BAdV-2 as a member of the genus Mastadenovirus.
Assuntos
Genoma Viral , Mastadenovirus/genética , Proteínas do Core Viral/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Bovinos , DNA Viral/análise , Dados de Sequência Molecular , Alinhamento de Sequência , Análise de Sequência de DNARESUMO
This paper describes a hypothesis on the origin of the members of the recently established adenovirus genus, Atadenovirus, invading cattle, sheep, deer, duck and poultry. Comparison of the phylogenetic trees of adenoviruses and their hosts suggests a very ancient but common origin for the atadenoviruses. The surprisingly large difference between these virus types and other adenoviruses infecting the same host can be easily understood by assuming their separate evolution in different hosts (e.g., in reptiles versus a coevolution with mammals and birds, respectively) followed by a later host switch.
Assuntos
Adenoviridae/genética , Evolução Molecular , Filogenia , Animais , Bovinos , Humanos , RépteisRESUMO
Similar open reading frames (ORF) overlapping the thymidine kinase (TK) gene on the opposite strand or being close to it have been identified in 8 herpesviruses (Jacobson et al., 1989). Study of the DNA sequence of BHV-1 TK gene published by Mittal and Field (1989) and Kit and Kit (1986) revealed the existence of a similar ORF in this virus genome, too.
Assuntos
DNA Viral/genética , Herpesvirus Bovino 1/genética , Timidina Quinase/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Herpesvirus Bovino 1/enzimologia , Dados de Sequência MolecularRESUMO
An idea is presented to use different letter types (e.g. L. 1, L, 1, L, l) for the alternating triplets coding the same amino acids. Such compact and demonstrative amino acid sequence comparisons can make easy the identification of differences at the DNA level if not seen in amino acid sequences (e.g. for choosing specific or non-specific primers for polymerase chain reaction).
Assuntos
Sequência de Aminoácidos , Códon/química , DNA/química , Sequência de Bases , Homologia de Sequência do Ácido NucleicoRESUMO
Physical maps of the DNA of bovine adenovirus (BAV) type 1 were established using ApaI, BamHI, BstEII, EcoRI and KpnI enzymes. The size of the viral genome was found to be around 35,000 base pairs. The orientation of the maps was determined by hybridizing Southern blots of BAV-1 DNA with the cloned hexon gene region of BAV-3.
Assuntos
Adenoviridae/genética , DNA Viral/análise , Infecções por Adenoviridae/microbiologia , Infecções por Adenoviridae/veterinária , Animais , Southern Blotting , Bovinos , Doenças dos Bovinos/microbiologia , Hibridização de Ácido Nucleico , Mapeamento por RestriçãoRESUMO
Molecular cloning of the HindIII fragments of bovine herpesvirus 1 (BHV-1) strain HB144, isolated from infectious bovine rhinotracheitis (IBR) in Hungary, and of an infectious pustular vulvovaginitis (IPV) reference strain (K22) is reported. So far 52% of the IBR viral genome and 28% of the IPV viral genome have been cloned. The analysis of differences between the strains is currently in progress.
Assuntos
DNA Viral/genética , Herpesvirus Bovino 1/genética , Rinotraqueíte Infecciosa Bovina/microbiologia , Animais , Bovinos , Clonagem Molecular , Hungria , Plasmídeos , Mapeamento por RestriçãoRESUMO
Physical maps of the genome of a bovine adenovirus (BAV) type 10 strain (isolate Belfast1) were constructed for eight restriction enzymes. The size of the viral genome was estimated to be around 29,800 base pairs (bp). The orientation of the maps was determined on the basis of partial DNA sequences of the cloned PstI/D fragment of Belfast1. This work is an introduction to DNA sequence and phylogenetic analysis of BAV-10 in order to clarify its taxonomic place.
Assuntos
Adenoviridae/classificação , Mapeamento Cromossômico/veterinária , DNA Viral/genética , Polimorfismo de Fragmento de Restrição , Adenoviridae/genética , Animais , Sequência de Bases , Bovinos , Enzimas de Restrição do DNA , DNA Viral/análise , DNA Viral/efeitos dos fármacos , Desoxirribonuclease BamHI , Desoxirribonuclease EcoRI , Genoma Viral , FilogeniaRESUMO
The DNA of a bovine herpesvirus 1 (BHV-1) strain isolated from calf encephalitis in Hungary was analysed with restriction enzymes. The cleavage pattern of the encephalitis strain Na/67 differed from those of all the other Hungarian BHV-1 isolates investigated so far. The EcoRI and HindIII cleavage patterns of virus strain Na/67 were found to be similar to the patterns of two other encephalitis strains (N569 and A663 from Australia and Argentina, respectively) characterized earlier. Strain Na/67 is the first isolate in Europe which showed the restriction enzyme pattern of BHV-1.3 previously supposed to be characteristic of encephalitis strains.
Assuntos
Doenças dos Bovinos/microbiologia , DNA Viral/análise , Encefalite/veterinária , Herpesvirus Bovino 1/genética , Animais , Bovinos , Encefalite/microbiologia , Hungria , Rinotraqueíte Infecciosa Bovina/microbiologia , Mapeamento por RestriçãoRESUMO
A primer pair and a polymerase chain reaction (PCR) method earlier tested for the detection of human and animal adenoviruses were used to demonstrate the presence of canine adenovirus type 1 (CAV-1) in tissue culture and clinical specimens. A simple procedure of sample preparation was elaborated making the PCR easily applicable in rapid confirmation of the diagnosis of infectious canine hepatitis.
Assuntos
Adenoviridae/isolamento & purificação , Hepatite Infecciosa Canina/diagnóstico , Animais , Linhagem Celular , Cães , Hepatite Infecciosa Canina/patologia , Humanos , Fígado/patologia , Fígado/virologia , MasculinoRESUMO
The authors report the data of the first survey on the incidence of post-weaning multisystemic wasting syndrome (PMWS) and porcine dermatitis and nephropathy syndrome (PDNS) in Hungary. A PCR method specific for the detection of porcine circovirus 2 (PCV-2) was developed, which proved to be suitable for diagnostic purposes. PCR screening of organ samples from pigs suspected to be affected with PMWS or PDNS revealed the presence of PCV-2 in 80% of the cases. Six PCV-2 genomes from Hungarian isolates were completely sequenced. Phylogenetic comparison with all the available PCV-2 sequences showed that porcine circoviruses circulating in Hungary are more variable than in several other European countries. Two Hungarian strains clustered together with the Spanish strains forming a distinct group; two others fell in a common group with the French, UK, and Dutch strains, whereas another two strains showed the closest relationship to two of the three known German PCV-2 sequences.
Assuntos
Infecções por Circoviridae/veterinária , Circovirus/genética , Doenças dos Suínos/epidemiologia , Doenças dos Suínos/virologia , Síndrome de Emaciação/veterinária , Animais , Infecções por Circoviridae/epidemiologia , Infecções por Circoviridae/virologia , Circovirus/isolamento & purificação , Primers do DNA , DNA Viral/análise , Genoma , Hungria/epidemiologia , Incidência , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase/veterinária , Prevalência , Suínos , Síndrome de Emaciação/epidemiologia , Síndrome de Emaciação/virologiaRESUMO
The full sequence of the fiber gene and partial sequence of the putative 17 kD protein gene of bovine adenovirus-2 (BAdV-2) were determined. The size of the fiber gene of BAdV-2 proved to be 561 amino acids, of which the amino acids 37 to 385 form a typical shaft domain of 22 repetitive motifs. On the complementary strand, a gene homologous to the 17 kD protein coded in the E4 region of several human adenoviruses was found. The sequence analysis seems to confirm the presence of an intron in the sequenced part of the E4 region.
Assuntos
Proteínas E4 de Adenovirus/genética , Proteínas do Capsídeo , Capsídeo/genética , Doenças dos Bovinos/virologia , Mastadenovirus/genética , Proteínas E4 de Adenovirus/química , Sequência de Aminoácidos , Animais , Sequência de Bases , Capsídeo/química , Bovinos , DNA Viral/química , Mastadenovirus/química , Dados de Sequência Molecular , Alinhamento de Sequência , Análise de Sequência de DNA , Homologia de Sequência de AminoácidosRESUMO
One of the plasmids present in a Haemophilus somnus strain isolated from nasal discharge of a cattle with respiratory disease was purified and cloned for DNA sequencing. The plasmid was found to be 1065 base pairs long with 39.2% G + C content, and showed no homology to any DNA sequenced so far. It has no capacity to code any protein longer than 43 residues. It is not clear yet if this plasmid codes Haemophilus somnus specific factors.
Assuntos
Doenças dos Bovinos/microbiologia , Infecções por Haemophilus/veterinária , Haemophilus/genética , Plasmídeos/genética , Animais , Sequência de Bases , Bovinos , Clonagem Molecular , Enzimas de Restrição do DNA/química , DNA Bacteriano/genética , Haemophilus/química , Infecções por Haemophilus/microbiologia , Dados de Sequência Molecular , Análise de Sequência de DNA , Homologia de Sequência do Ácido NucleicoRESUMO
At abattoirs and farms, 1248 sera were collected from animals representing 121 farms, and examined by complement fixation test using Mycoplasma mycoides subspecies mycoides small colony type (MmmSC) antigen. All sera were negative except seven from four farms, giving ++ reactions in the serum dilution of 1:10. On retesting, these sera and additional 30 sera collected repeatedly in both farms gave negative results. In isolation attempts, 953 lung samples collected from slaughtered cattle at the same abattoirs, and 326 nasal swabs collected from 11 herds proved to be negative for the presence of MmmSC, but M. bovis was isolated frequently. In the small farms 23.95% of the animals had pleurisy and/or pneumonia while in the large herds 34.69% had lesions. DNA extracted from 50 nasal swabs and 430 lung samples was examined by polymerase chain reaction (PCR) using M. mycoides cluster-specific primers. DNA from further 325 lung samples was tested by the more specific M. mycoides subspecies mycoides small colony/large colony/capri specific primers and 196 samples by nested PCR specific for MmmSC. All gave negative results. The detection level of cluster-specific primers and the more specific primers was 33.4 pg of DNA, whereas that of nested PCR was 0.33 pg.
Assuntos
Doenças dos Bovinos/diagnóstico , Mycoplasma mycoides/isolamento & purificação , Pleuropneumonia Contagiosa/diagnóstico , Animais , Bovinos , Doenças dos Bovinos/epidemiologia , Testes de Fixação de Complemento , Hungria/epidemiologia , Pulmão/microbiologia , Pleuropneumonia Contagiosa/epidemiologia , Reação em Cadeia da PolimeraseRESUMO
Plant pathogens can be divided into biotrophs and necrotrophs according to their different life styles; biotrophs prefer living, while necrotrophs prefer dead cells for nutritional purposes. Therefore tissue necrosis caused by reactive oxygen species (ROS) during pathogen infection increases host susceptibility to necrotrophic, but resistance to biotrophic pathogen. Consequently, elevation of antioxidant capacity of plants enhances their tolerance to development of necroses caused by necrotrophic pathogens. Plant hormones can strongly influence induction of ROS and antioxidants, thereby influencing susceptibility or resistance of plants to pathogens. Pathogen-induced ROS themselves are considered as signaling molecules. Generally, salicylic acid (SA) signaling induces defense against biotrophic pathogens, whereas jasmonic acid (JA) against necrotrophic pathogens. Furthermore pathogens can modify plant's defense signaling network for their own benefit by changing phytohormone homeostasis. On the other hand, ROS are harmful also to the pathogens, consequently they try to defend themselves by elevating antioxidant activity and secreting ROS scavengers in the infected tissue. The Janus face nature of ROS and plant cell death on biotrophic and on necrotrophic pathogens is also supported by the experiments with BAX inhibitor-1 and the mlo mutation of Mlo gene in barley. It was found that ROS and elevated plant antioxidant activity play an important role in systemic acquired resistance (SAR) and induced systemic resistance (ISR), as well as in mycorrhiza induced abiotic and biotic stress tolerance of plants.