RESUMEN
In 1983, Mycoplasma sp. strain 1220 was isolated in Hungary from the phallus lymph of a gander with phallus inflammation. Between 1983 and 2017, Mycoplasma sp. 1220 was also identified and isolated from the respiratory tract, liver, ovary, testis, peritoneum and cloaca of diseased geese in several countries. Seventeen studied strains produced acid from glucose and fructose but did not hydrolyse arginine or urea, and all grew under aerobic, microaerophilic and anaerobic conditions at 35 to 37 ËC in either SP4 or pleuropneumonia-like organism medium supplemented with glucose and serum. Colonies on agar showed a typical fried-egg appearance and transmission electron microscopy revealed a typical mycoplasma cellular morphology. Molecular characterization included analysis of the following genetic loci: 16S rRNA, 23S rRNA, 16S-23S rRNA ITS, rpoB, rpoC, rpoD, uvrA, parC, topA, dnaE, fusA and pyk. The genome was sequenced for type strain 1220T. The 16S rRNA gene sequences of studied strains of Mycoplasma sp. 1220 shared 99.02-99.19â% nucleotide similarity with M. anatis strains but demonstrated ≤95.00-96.70â% nucleotide similarity to the 16S rRNA genes of other species of the genus Mycoplasma. Phylogenetic, average nucleotide and amino acid identity analyses revealed that the novel species was most closely related to Mycoplasma anatis. Based on the genetic data, we propose a novel species of the genus Mycoplasma, for which the name Mycoplasma anserisalpingitidis sp. nov. is proposed with the type strain 1220T (=ATCC BAA-2147T=NCTC 13513T=DSM 23982T). The G+C content is 26.70 mol%, genome size is 959110 bp.
Asunto(s)
Gansos/microbiología , Mycoplasma/clasificación , Filogenia , Animales , Técnicas de Tipificación Bacteriana , Composición de Base , ADN Bacteriano/genética , Femenino , Genes Bacterianos , Hungría , Mycoplasma/aislamiento & purificación , ARN Ribosómico 16S/genética , ARN Ribosómico 23S/genética , Análisis de Secuencia de ADNRESUMEN
A large percentage of primary sensory neurons in the trigeminal ganglia (TG) contain neuropeptides such as tachykinins or calcitonin gene-related peptide. Neuropeptides released from the central terminals of primary afferents sensitize the secondary nociceptive neurons in the trigeminal nucleus caudalis (TNC), but also activate glial cells contributing to neuroinflammation and consequent sensitization in chronic orofacial pain and migraine. In the present study, we investigated the newest member of the tachykinin family, hemokinin-1 (HK-1) encoded by the Tac4 gene in the trigeminal system. HK-1 had been shown to participate in inflammation and hyperalgesia in various models, but its role has not been investigated in orofacial pain or headache. In the complete Freund's adjuvant (CFA)-induced inflammatory orofacial pain model, we showed that Tac4 expression increased in the TG in response to inflammation. Duration-dependent Tac4 upregulation was associated with the extent of the facial allodynia. Tac4 was detected in both TG neurons and satellite glial cells (SGC) by the ultrasensitive RNAscope in situ hybridization. We also compared gene expression changes of selected neuronal and glial sensitization and neuroinflammation markers between wild-type and Tac4-deficient (Tac4-/-) mice. Expression of the SGC/astrocyte marker in the TG and TNC was significantly lower in intact and saline/CFA-treated Tac4-/- mice. The procedural stress-related increase of the SGC/astrocyte marker was also strongly attenuated in Tac4-/- mice. Analysis of TG samples with a mouse neuroinflammation panel of 770 genes revealed that regulation of microglia and cytotoxic cell-related genes were significantly different in saline-treated Tac4-/- mice compared to their wild-types. It is concluded that HK-1 may participate in neuron-glia interactions both under physiological and inflammatory conditions and mediate pain in the trigeminal system.
Asunto(s)
Dolor Facial/etiología , Regulación de la Expresión Génica , Taquicininas/genética , Ganglio del Trigémino/metabolismo , Animales , Biomarcadores , Modelos Animales de Enfermedad , Susceptibilidad a Enfermedades , Dolor Facial/metabolismo , Dolor Facial/fisiopatología , Técnica del Anticuerpo Fluorescente , Perfilación de la Expresión Génica , Hiperalgesia , Macrófagos/metabolismo , Ratones , Ratones Noqueados , Neuroglía/metabolismo , Células Receptoras Sensoriales/metabolismo , Taquicininas/metabolismo , Neuralgia del Trigémino/etiología , Neuralgia del Trigémino/metabolismoRESUMEN
Autologous blood derived products, such as platelet-rich plasma (PRP) and platelet-rich fibrin (PRF) are widely applied in regenerative therapies, in contrast to the drawbacks in their application, mainly deriving from the preparation methods used. Eliminating the disadvantages of both PRP and PRF, hyperacute serum (HAS) opens a new path in autologous serum therapy showing similar or even improved regenerative potential at the same time. Despite the frequent experimental and clinical use of PRP and HAS, their protein composition has not been examined thoroughly yet. Thus, we investigated and compared the composition of HAS, serum, PRP and plasma products using citrate and EDTA by simple laboratory tests, and we compared the composition of HAS, serum, EDTA PRP and plasma by Proteome Profiler and ELISA assays. According to our results the natural ionic balance was upset in both EDTA and citrate PRP as well as in plasma. EDTA PRP contained significantly higher level of growth factors and cytokines, especially platelet derived angiogenic and inflammatory proteins, that can be explained by the significantly higher number of platelets in EDTA PRP. The composition analysis of blood derivatives revealed that although the preparation method of PRP and HAS were similar, the ionic and protein composition of HAS could be advantageous for cell function.
Asunto(s)
Plasma Rico en Plaquetas , Suero , Proteínas Sanguíneas/química , Fraccionamiento Químico , Humanos , Fibrina Rica en Plaquetas , Plasma Rico en Plaquetas/química , Suero/químicaRESUMEN
OBJECTIVE: We report the infection of New Zealand white rabbits with Epstein-Barr virus (EBV). METHODS: EBV prepared in B95-8 (producer) cells was inoculated to rabbits by combined intranasal and oral routes. Blood and white blood cell (WBC) samples were taken before infection, then on days 8, 28 and 98 post-infection (p.i.). RESULTS: Administration of either 3 × 10(8) (group A, 11 rabbits) or 1 × 10(9) (group B, 10 rabbits) EBV DNA copies per animal induced subacute and/or persistent infection. The IgG antibodies in plasma were detected by ELISA as well as by immunoblot (IB). The IB bands showed mainly antibodies to the BZRF1/Zta transactivation polypeptide (69.2%), the p54 early protein (53.4%) and to the p23 capsid protein (35.8%). No anti-EBNA1 antibody was detected throughout. Viral DNA could be detected by PCR in WBCs and/or spleen of 7 out of 21 infected rabbits (30%), while 60-80% of them showed serologic response. The transiently present EBV DNA was accompanied by LMP1 antigen. CONCLUSIONS: Rabbits developed persistent EBV infection in the absence of EBNA1 antibodies and by the lack of typical infectious mononucleosis-like syndrome. The absence of EBNA1 antibody may reflect the lack of EBNA1 in B cells of EBV-inoculated rabbits.
Asunto(s)
Modelos Animales de Enfermedad , Infecciones por Virus de Epstein-Barr/patología , Infecciones por Virus de Epstein-Barr/virología , Herpesvirus Humano 4/crecimiento & desarrollo , Leucocitos Mononucleares/patología , Animales , Anticuerpos Antivirales/sangre , Ensayo de Inmunoadsorción Enzimática , Herpesvirus Humano 4/inmunología , Humanos , Immunoblotting , Inmunoglobulina G/sangre , Mononucleosis Infecciosa/patología , ConejosRESUMEN
The synonymous codon usage pattern of African swine fever virus (ASFV), the similarity degree of the synonymous codon usage between this virus and some organisms and the synonymous codon usage bias for the translation initiation region of viral functional genes in the whole genome of ASFV have been investigated by some simply statistical analyses. Although both GC12% (the GC content at the first and second codon positions) and GC3% (the GC content at the third codon position) of viral functional genes have a large fluctuation, the significant correlations between GC12 and GC3% and between GC3% and the first principal axis of principle component analysis on the relative synonymous codon usage of the viral functional genes imply that mutation pressure of ASFV plays an important role in the synonymous codon usage pattern. Turning to the synonymous codon usage of this virus, the codons with U/A end predominate in the synonymous codon family for the same amino acid and a weak codon usage bias in both leading and lagging strands suggests that strand compositional asymmetry does not take part in the formation of codon usage in ASFV. The interaction between the absolute codon usage bias and GC3% suggests that other selections take part in the formation of codon usage, except for the mutation pressure. It is noted that the similarity degree of codon usage between ASFV and soft tick is higher than that between the virus and the pig, suggesting that the soft tick plays a more important role than the pig in the codon usage pattern of ASFV. The translational initiation region of the viral functional genes generally have a strong tendency to select some synonymous codons with low GC content, suggesting that the synonymous codon usage bias caused by translation selection from the host takes part in modulating the translation initiation efficiency of ASFV functional genes.
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Virus de la Fiebre Porcina Africana/genética , Fiebre Porcina Africana/virología , Codón , Iniciación de la Cadena Peptídica Traduccional , Proteínas Virales/genética , Virus de la Fiebre Porcina Africana/clasificación , Animales , Composición de Base , Datos de Secuencia Molecular , PorcinosRESUMEN
We performed a comparative study on the development of mastitis induced by Mycoplasma arginini or Streptococcus dysgalactiae after challenging the cows. Mycoplasma arginini did not cause any clinical symptoms on its own, resulting in only a transient increase of somatic cell count (SCC; increase ranging from 0.5 × 10(6) to 0.8 × 10(6) cells/mL) and a slight decrease of milk production (10%) for 5 d. In contrast, Strep. dysgalactiae induced more severe clinical signs in animals and SCC increased to 1.60 × 10(6) to 2.11 × 10(6) cells/mL for 10 d. In addition, milk production decreased (22.9 to 27.0%) for 10 d. After 3 mo (2 mo after the first challenge), animals that were challenged previously with M. arginini were rechallenged with Strep. dysgalactiae. Severe clinical mastitis developed, with very high SCC (5.00 × 10(6) to 21.5 × 10(6) cells/mL), and a very significant reduction of milk production (28.6 to 68.7%), which lasted more than 4 wk, was observed. The severe clinical mastitis developed not only in cows inoculated with Strep. dysgalactiae andM. arginini in the same udder quarter but also in cows infected in the quarter previously not challenged with mycoplasma. Cows challenged first with Strep. dysgalactiae and rechallenged with M. arginini 2 mo later developed only slight changes in both SCC and milk production, similar to those when the cows were challenged with M. arginini alone. We conclude that M. arginini infection does not cause remarkable mastitis (characterized by decrease in milk production and increase of SCC) but it significantly predisposes animals to infection with Strep. dysgalactiae, leading to severe clinical mastitis.
Asunto(s)
Mastitis Bovina/microbiología , Infecciones por Mycoplasma/veterinaria , Mycoplasma , Infecciones Estreptocócicas/veterinaria , Streptococcus , Animales , Bovinos , Coinfección/microbiología , Coinfección/veterinaria , Femenino , Lactancia , Leche/citología , Infecciones por Mycoplasma/microbiologíaRESUMEN
Eosinophilic fasciitis (EF) with generalized sclerodermiform skin lesions developed over a 19-month period in a previously healthy 23-year-old man. Although we confirmed EF by skin histology and laboratory tests, the recurrent fevers and the clinical observation of sclerotic prepuce with urethritis indicated further bacteriological analysis by conventional microbiological and DNA-based tests. Urethra cultures were positive for an arginine-hydrolyzing mycoplasma and Ureaplasma urealyticum. The patient also had serum IgM antibodies to Mycoplasma pneumoniae using enzyme-linked immunosorbent assay (ELISA)-based qualitative detection. Mycoplasma arginini was isolated from two independent venous blood serum samples and was identified by conventional microbiological tests and sequencing of the 16S rRNA and rpoB genes (GenBank sequence accession numbers HM179555 and HM179556, respectively). M. arginini genomic DNA also was detected by species-specific PCR in the skin lesion biopsy sample. Treatment with corticosteroids and long-term courses of selected antibiotics led to remission of skin symptoms and normalization of laboratory values. This report provides the first evidence of EF associated with mycoplasma infection and the second report of human infection with M. arginini and therefore suggests that this mycoplasma infection might have contributed to the pathogenesis of the disease.
Asunto(s)
Eosinofilia/diagnóstico , Fascitis/diagnóstico , Fascitis/microbiología , Infecciones por Mycoplasma/complicaciones , Mycoplasma/aislamiento & purificación , Enfermedades Cutáneas Bacterianas/complicaciones , Bacteriemia/diagnóstico , Bacteriemia/microbiología , Bacteriemia/patología , Técnicas de Tipificación Bacteriana , Biopsia , Sangre/microbiología , Análisis por Conglomerados , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Eosinofilia/complicaciones , Eosinofilia/patología , Fascitis/complicaciones , Fascitis/patología , Histocitoquímica , Humanos , Masculino , Datos de Secuencia Molecular , Mycoplasma/clasificación , Infecciones por Mycoplasma/microbiología , Infecciones por Mycoplasma/patología , Filogenia , Reacción en Cadena de la Polimerasa , ARN Ribosómico 16S/genética , Recurrencia , Análisis de Secuencia de ADN , Enfermedades Cutáneas Bacterianas/microbiología , Enfermedades Cutáneas Bacterianas/patología , Adulto JovenRESUMEN
Mycoplasma infection is still very common in chicken and turkey flocks. Several low-pathogenicity avian influenza (LPAI) viruses are circulating in wild birds that can be easily transmitted to poultry flocks. However, the effect of LPAI on mycoplasma infection is not well understood. The aim of the present study was to investigate the infection of LPAI virus H3N8 (A/mallard/Hungary/19616/07) in chickens challenged with Mycoplasma gallisepticum. Two groups of chickens were aerosol challenged with M. gallisepticum. Later one of these groups and one mycoplasma-free group were aerosol challenged with the LPAI H3N8 virus. The birds were observed for clinical signs for 8 days, then euthanized, and examined for the presence of M. gallisepticum in the trachea, lung, air sac, liver, spleen, kidney and heart, and for developing anti-mycoplasma and anti-viral antibodies. The LPAI H3N8 virus did not cause any clinical signs but M. gallisepticum infection caused clinical signs, reduction of body weight gain and colonization of the inner organs. These parameters were more severe in the birds co-infected with M. gallisepticum and LPAI H3N8 virus than in the group challenged with M. gallisepticum alone. In addition, in the birds infected with both M. gallisepticum and LPAI H3N8 virus, the anti-mycoplasma antibody response was reduced significantly when compared with the group challenged with M. gallisepticum alone. Co-infection with LPAI H3N8 virus thus enhanced pathogenesis of M. gallisepticum infection significantly.
Asunto(s)
Pollos , Coinfección/veterinaria , Subtipo H3N8 del Virus de la Influenza A/patogenicidad , Gripe Aviar/microbiología , Infecciones por Mycoplasma/veterinaria , Mycoplasma gallisepticum , Enfermedades de las Aves de Corral/microbiología , Enfermedades de las Aves de Corral/virología , Animales , Anticuerpos Antibacterianos/sangre , Anticuerpos Antivirales/sangre , Peso Corporal , Coinfección/microbiología , Coinfección/virología , Infecciones por Mycoplasma/virología , Pruebas Serológicas/veterinaria , Vísceras/microbiologíaRESUMEN
An alternative vaccine design approach and diagnostic kits are highly required against the anticipated pandemicity caused by the South African Territories type 2 (SAT2) Foot and Mouth Disease Virus (FMDV). However, the distinct antigenicity and immunogenicity of VP1, VP0, and VP3 of FMDV serotype SAT2 are poorly understood. Similarly, the particular roles of the three structural proteins in novel vaccine design and development remain unexplained. We therefore constructed VP1, VP0, and VP3 encoding gene (SAT2:JX014256 strain) separately fused with His-SUMO (histidine-small ubiquitin-related modifier) inserted into pET-32a cassette to express the three recombinant proteins and separately evaluated their antigenicity and immunogenicity in mice. The fusion protein was successfully expressed and purified by the Ni-NTA resin chromatography. The level of serum antibody, spleen lymphocyte proliferation, and cytokines against the three distinct recombinant proteins were analyzed. Results showed that the anti-FMDV humoral response was triggered by these proteins, and the fusion proteins did enhance the splenocyte immune response in the separately immunized mice. We observed low variations among the three fusion proteins in terms of the antibody and cytokine production in mice. Hence, in this study, results demonstrated that the structural proteins of SAT2 FMDV could be used for the development of immunodiagnostic kits and subunit vaccine designs.
Asunto(s)
Proteínas de la Cápside/genética , Escherichia coli/genética , Virus de la Fiebre Aftosa/genética , Virus de la Fiebre Aftosa/inmunología , Fiebre Aftosa/prevención & control , Inmunogenicidad Vacunal , Proteínas Estructurales Virales/genética , Vacunas Virales/inmunología , Animales , Proteínas de la Cápside/inmunología , Femenino , Fiebre Aftosa/inmunología , Fiebre Aftosa/virología , Virus de la Fiebre Aftosa/química , Inmunización , Ratones , Ratones Endogámicos BALB C , Proteínas Recombinantes/genética , Sudáfrica , Organismos Libres de Patógenos Específicos , Proteínas Estructurales Virales/clasificación , Proteínas Estructurales Virales/inmunología , Vacunas Virales/genéticaRESUMEN
Mycoplasma synoviae (MS) is a poultry pathogen with a reported distribution throughout the world. Vaccination is utilized as an important component of MS control programs for MS infection. The aim of this study was to evaluate protection efficacy of an inactivated MS vaccine (MS bacterin) with different adjuvants in broilers against a Chinese field isolate (CHN-BZJ2-2015). Vaccination with adjuvants ISA 71 VG and chitosan, respectively, enhanced specific lymphocyte proliferation responses and upregulated the expression of IL-1ß, IL-6, IL-2 and IFN-γ prior to challenge. Furthermore, vaccination with adjuvant ISA 71 VG elicited the highest antibody titers, exhibited significantly lower air sac, foot pad and tracheal lesions than the other groups (P < 0.05), and decreased MS colonization. These results demonstrated that inactivated MS vaccine with ISA 71 VG is able to induce both cellular and humoral immune response in broilers and confers a high level of protection upon challenge, demonstrating a potential application in the field.
Asunto(s)
Adyuvantes Inmunológicos/administración & dosificación , Anticuerpos Antibacterianos/sangre , Vacunas Bacterianas/inmunología , Infecciones por Mycoplasma/veterinaria , Enfermedades de las Aves de Corral/prevención & control , Adyuvantes Inmunológicos/clasificación , Animales , Proliferación Celular , Pollos/inmunología , Quitosano/administración & dosificación , Quitosano/inmunología , Citocinas/genética , Citocinas/inmunología , Inmunidad Celular , Inmunidad Humoral , Infecciones por Mycoplasma/prevención & control , Mycoplasma synoviae/inmunología , Enfermedades de las Aves de Corral/microbiología , Vacunación/veterinaria , Vacunas de Productos Inactivados/administración & dosificación , Vacunas de Productos Inactivados/inmunologíaRESUMEN
OBJECTIVES: In this study, the genetic diversity, phylogenetic grouping, antimicrobial resistance and extended-spectrum ß-lactamase (ESBL) types of Escherichia coli isolates from chickens, dogs, pigs and yaks in six prefectures of Gansu and Qinghai Provinces, China, were investigated. METHODS: E. coli was isolated from diarrhoeic and healthy faecal samples. Multilocus sequence typing (MLST), phylogenetic grouping, antimicrobial resistance and ESBL profiles were investigated. RESULTS: A total of 142 MLST sequence types (STs) were identified from 400 E. coli isolates. eBURST clustering analysis resolved the 142 STs into 19 clonal complexes (CCs) and 67 singletons. PCR phylogenetic typing determined the isolation rate of potentially pathogenic B2/D group isolates among all E. coli to be 12.5% from healthy animal samples and 17.5% from diarrhoeic samples. Antimicrobial susceptibility testing revealed 78 antimicrobial resistance patterns. E. coli resistance rates were highest to doxycycline, ampicillin and tetracycline, whereas polymyxin B and meropenem had the lowest resistance rates. All polymyxin B-resistant E. coli isolates were positive for the mcr-1 gene. A total of 62 ESBL-producing isolates were identified. The ESBL prevalence was 55.0% in diarrhoeic samplings and 5.6% in healthy animals. TEM (82.3%) was the predominant ESBL type, followed by CTM (43.5%) and SHV (19.4%). CONCLUSION: E. coli isolates in the study area have a high diversity of genetic and antimicrobial resistance patterns but a relatively low isolation rate of potentially pathogenic phylogroups. However, the somewhat high isolation rate of multidrug-resistant E. coli, particularly ESBL-producing isolates, requires continual surveillance of E. coli from animals in these areas.
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Infecciones por Escherichia coli , Proteínas de Escherichia coli , Escherichia coli , Animales , Antibacterianos/farmacología , Bovinos , Pollos , China/epidemiología , Perros , Farmacorresistencia Bacteriana/efectos de los fármacos , Escherichia coli/efectos de los fármacos , Escherichia coli/genética , Infecciones por Escherichia coli/epidemiología , Infecciones por Escherichia coli/veterinaria , Proteínas de Escherichia coli/genética , Variación Genética , Tipificación de Secuencias Multilocus , Filogenia , Porcinos , beta-Lactamasas/genéticaRESUMEN
One option to fight joint degradation and inflammation in osteoarthritis is the injection of activated blood products into the synovial space. It has been demonstrated that hyperacute serum is the most proliferative among plasma products, so we investigated how the cytokine milieu of osteoarthritic knee joint reacts to hyperacute serum treatment in vitro. Cartilage, subchondral bone, and synovial membrane explanted from osteoarthritic knees were stimulated by interleukin-1 beta (IL-1ß) and the concentration of 39 biomarkers was measured in the co-culture supernatant after hyperacute serum treatment. The IL-1ß stimulation triggered a strong inflammatory response and enhanced the concentrations of matrix metalloproteinase 3 and 13 (MMP-3 and MMP-13), while hyperacute serum treatment reduced inflammation by decreasing the concentrations of IL-1ß, tumor necrosis factor alpha (TNF-α), interleukin-6 receptor alpha (IL-6Rα), and by increasing the level of interleukin-1 antagonist (IL-1RA) Cell viability increased by day 5 in the presence of hyperacute serum. The level of MMPs-1, 2, and 9 were higher on day 3, but did not increase further until day 5. The concentrations of collagen 1 alpha 1 (COL1A1) and osteonectin were increased and receptor activator of nuclear factor kappa-B ligand (RANKL) was reduced in response to hyperacute serum. We concluded that hyperacute serum treatment induces cell proliferation of osteoarthritic joint tissues and affects the cytokine milieu towards a less inflamed state.
Asunto(s)
Citocinas/metabolismo , Interleucina-1beta/farmacología , Articulación de la Rodilla/efectos de los fármacos , Articulación de la Rodilla/metabolismo , Osteoartritis de la Rodilla/terapia , Adulto , Cartílago/efectos de los fármacos , Cartílago/patología , Técnicas de Cocultivo , Femenino , Humanos , Articulación de la Rodilla/patología , Masculino , Persona de Mediana Edad , Membrana Sinovial/efectos de los fármacos , Técnicas de Cultivo de Tejidos , Adulto JovenRESUMEN
Recently, it was demonstrated using in vitro assays that the avian pathogen Mycoplasma gallisepticum is able to invade nonphagocytic cells. It was also shown that this mycoplasma can survive and multiply intracellularly for at least 48 h and that this cell invasion capacity contributes to the systemic spread of M. gallisepticum from the respiratory tract to the inner organs. Using the gentamicin invasion assay and a differential immunofluorescence technique combined with confocal laser scanning microscopy, we were able to demonstrate in in vitro experiments that M. gallisepticum is also capable of invading sheep and chicken erythrocytes. The frequencies of invasion of three well-defined M. gallisepticum strains were examined over a period of 24 h, and a significant increase in invasiveness occurred after 8 h of infection. In addition, blood samples derived from chickens experimentally infected via the aerosol route with the virulent strain M. gallisepticum R(low) were analyzed. Surprisingly, M. gallisepticum R(low) was detected in the bloodstream of infected chickens by nested PCR, as well as by differential immunofluorescence and interference contrast microscopy that showed that mycoplasmas were not only on the surface but also inside chicken erythrocytes. This finding provides novel insight into the pathomechanism of M. gallisepticum and may have implications for the development of preventive strategies.
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Pollos/microbiología , Eritrocitos/microbiología , Infecciones por Mycoplasma/veterinaria , Mycoplasma gallisepticum/fisiología , Enfermedades de las Aves de Corral/microbiología , Animales , Eritrocitos/ultraestructura , Células HeLa , Humanos , Infecciones por Mycoplasma/microbiología , Mycoplasma gallisepticum/ultraestructuraRESUMEN
INTRODUCTION: Despite overwhelming experimental work, there are no licensed vaccines against the most frequent Alphaherpesviruses, namely herpes simplex virus 1 and 2 (HSV1 and 2) nor against the Epstein-Barr virus (EBV), a member of the subfamily Gammaherpesvirus. AREAS COVERED: Since the DNAs of both HSVs reside in the regional sensory ganglia in a latent state (i.e. as circularized episomal molecules), a corresponding vaccine might be useful for immunotherapy rather than for prevention of primary infection. Here we describe the design of a purified subunit vaccine as well as the preparation and efficacy of a recombinant fusion protein consisting of the gD ectodomain from our domestic attenuated HSV1 strain HSZP. The EBV vaccines considered so far, were destined for prevention of infectious mononucleosis (IM) or to prevent formation of EBV related tumors. To design the EBV peptide vaccine, at least 15 carefully selected immunogenic epitopes coming from 12 virus coded proteins were bound to synthetic micro-particle carriers along with a non-specific pathogen recognizing receptor (PRR) stimulating both the T as well as B lymphocytes. EXPERT COMMENTARY: The efficacy of a novel EBV peptide in the rabbit model was based on criteria such as antibody formation (EA-D detected by ELISA, early and capsid proteins tested by immunoblot), presence of LMP1 antigen and of viral DNA in peripheral white blood cells. Out of 19 peptide combinations used for vaccination, at least 6 showed a satisfactory protective effect.
Asunto(s)
Infecciones por Virus de Epstein-Barr/prevención & control , Herpes Simple/prevención & control , Vacunas contra Herpesvirus/administración & dosificación , Animales , Infecciones por Virus de Epstein-Barr/inmunología , Herpes Simple/inmunología , Vacunas contra el Virus del Herpes Simple/administración & dosificación , Vacunas contra el Virus del Herpes Simple/inmunología , Herpesvirus Humano 1/inmunología , Herpesvirus Humano 2/inmunología , Herpesvirus Humano 4/inmunología , Vacunas contra Herpesvirus/inmunología , Humanos , ConejosRESUMEN
Viral infections trigger the innate immune system to produce interferons (IFNs), which play important role in host antiviral responses. Co-evolution of viruses with their hosts has favored development of various strategies to evade the effects of IFNs, enabling viruses to survive inside host cells. One such strategy involves inhibition of IFN signaling pathways by non-structural proteins. In this review, we provide a brief overview of host signaling pathways inducing IFN production and their suppression by picornavirus non-structural proteins. Using this strategy, picornaviruses can evade the host immune response and replicate inside host cells.
RESUMEN
The number of multidrug-resistant strains of Riemerella anatipestifer continues to increase, and new strategies for the treatment of associated infections are necessary. Recently, numerous studies have shown that efflux pumps (EPs) play key roles in universal bacterial mechanisms that contribute to antibiotic resistance. In addition, studies have shown that the effects of antibiotics that are subjected to efflux can be reinforced by their combined use with efflux pump inhibitors (EPIs). Unfortunately, the role of the efflux system in R. anatipestifer remains barely understood. In this study, we evaluated the role of EPs and resistance genes in the resistance generated by clinical strains of R. anatipestifer to antibiotics. A set of 10 R. anatipestifer strains were characterized by drug resistance, associated resistance genes, and antibiotic profiles in the presence and absence of EPIs. Efflux activity was studied on a real time basis through a fluorometric method. Quantification of the levels of mRNA transcription of efflux pump genes (EPGs) was determined by RT-qPCR. Several approaches (detection of resistance genes, drug susceptibility testing, and growth kinetics analysis) were used to assess the correlation between the effect of the EPIs and the resistance levels. Analysis of the R. anatipestifer growth inhibition tests showed that the antibiotic activity was enhanced by the synergy of EPIs. Among the various resistance genes that confer antibiotic resistance, different minimum inhibitory concentrations (MICs) were observed. The different levels of resistance were reduced by EPIs. Real time fluorometry showed that all the R. anatipestifer strains presented inherent efflux activity, conferring varying levels of inhibition in the presence of EPIs. Moreover, 15 EPGs were overexpressed in the presence of antibiotics. The addition of EPIs to antibiotics led to downregulation in the expression of some EPGs and a simultaneous increase in drug resistance and sensitivity. These results demonstrated the contribution of these EPs in the resistant phenotype of the clinical strains of R. anatipestifer that are under investigation, independently of the resistant genotype of the respective strains. Intrinsic efflux activity was possibly linked to the evolution of resistance in multidrug-resistant isolates of R. anatipestifer. Furthermore, the inhibition of EPs by EPIs could enhance the clinical effects of antibiotics.
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The capture of mycoplasmas (M. hominis, M. buccale, M. fermentans, M. bovis, M. synoviae, M. gallisepticum and M. arthritidis) based on lipid structures and adhesion molecules present in the mycoplasmal membrane was tested using different chromatographic resins (ActiClean Etox, ClarEtox, Heparin-Actigel, Sulfated Hiflow and SulfEtox). All of the resins efficiently reduced mycoplasma concentrations in Phosphate Buffered Saline (PBS) and in Fetal Bovine Serum (FBS) by 3-8 logs in a few minutes. This technology could be used for removing mycoplasmas from tissue culture components such as serum, and for concentrating mycoplasmas in vaccine production.
Asunto(s)
Adhesión Bacteriana/fisiología , Mycoplasma/aislamiento & purificación , Mycoplasma/fisiología , Resinas Sintéticas , Adsorción , Moléculas de Adhesión Celular/fisiología , Membrana Celular/fisiología , CromatografíaRESUMEN
The pathogenesis of sepsis begins with the proliferation of micro-organisms at a site of infection, followed by invasion of the bloodstream and other organs. Gram-negative bacteria account for a large part of sepsis cases. The structural component of Gram-negative bacteria, endotoxin or lipopolysaccharide (LPS), induces the synthesis and release of endogenous mediators of sepsis. A growing number of investigations of the molecular mechanisms occurring in sepsis, point to endotoxin as a central mediator leading to multi-organ failure and death. In numerous clinical trials, attempts to target molecules downstream of endotoxin have been made, but have not been associated with improved survival. We describe an affinity-based system for the selective removal of endotoxin from plasma. The small-scale device, a 1.5 ml cartridge, contains beads that bind endotoxin with high specificity and efficiency. In addition, evidence is presented that this device does not affect plasma hemostasis, nor does it activate the complement system. Taken together, these results represent a proof of principle for endotoxin removal from plasma, which may be of clinical value to treat sepsis by extracorporeal circulation of the blood through a scaled-up version of this endotoxin-removing device.
Asunto(s)
Endotoxinas/sangre , Diálisis Renal/métodos , Sepsis/terapia , Pruebas de Coagulación Sanguínea , Cromatografía de Afinidad/métodos , Endotoxinas/aislamiento & purificación , Ensayo de Inmunoadsorción Enzimática , Escherichia coli , Humanos , Pseudomonas , Sepsis/sangre , Factor de Necrosis Tumoral alfa/análisisRESUMEN
The Epstein-Barr virus (Human herpesvirus 4) encodes approximately 80 proteins, from which 15 possess at least 90 antigenic epitopes. Many of them stimulate the T cell receptors (TCR), but a few interact with the B cell receptors (BCR). Activation of B-cells and subsequent antibody production has not only been related to at least 3 envelope glycoproteins (mostly gp350) but also to latency associated membrane proteins (LMPs). The majority of EBV epitopes (over 80) inducing either cytotoxic and/or helper T lymphocytes were located on non-structural and/or latency associated polypeptides. The former interaction mediated by CD8plus/T cells is restricted by the HLA I molecules, predominantly of HLA-A subclass. In acute infectious mononucleosis (IM) patients (about 40 %) a considerable proportion of HLA B8 restricted CTL reactivity is directed against a single peptide (RAKFKQLL) of transactivator protein BZLF1/Zta. The EBV vaccines designed so far fall into two categories: those preventing any kind of infection (including prophylaxis of EBVassociated malignancies) and those designed for therapeutic purposes (to be used in subjects already infected). Preventive vaccines protecting against acute disease (such as IM) contain, as a rule, the gp350 polypeptide(s) encoded by the BLLF1 gene. Vaccines destined for tumor prevention rather consist of peptides derived from latency associated nuclear proteins (EBNA 2, 3 and 6) and/or from oncogenic latent membrane proteins (LMP1/LMP2a). Whereas the former generates antibodies preventing virus entry, the latter would potentiate the cell mediated response. In addition to recently described and purified individual recombinant immunogenic EBV polypeptides and/or their mixes, new perspectives were opened by construction of random overlapping strongly immunogenic scrambled polypeptide(s). Further novel approaches are based on carefully selected antigenic peptides (oligopeptides) coming from both, structural as well as non-structural or latencyassociated proteins bound to suitable carriers. Any constructs based on latency-associated proteins might be useful either for immunoprophylactic therapy following bone marrow and/or heart transplantations or for the prevention of EBVrelated tumors such as lymphomas and nasopharyngeal carcinoma. Due to the growing importance of the selected immunogenic epitopes as future vaccine components, at least the half of them has been patented not only as the natural amino acid sequence but also in different variations.
Asunto(s)
Antígenos Virales/inmunología , Epítopos/inmunología , Herpesvirus Humano 4/inmunología , Proteínas Virales/inmunología , Vacunas Virales/inmunología , HumanosRESUMEN
Although there is a significant effort in the design of a selective CDK9/CycT1 inhibitor, no compound has been proven to be a specific inhibitor of this kinase so far. The aim of this research was to develop novel and selective phosphorus containing CDK9/CycT1 inhibitors. Molecules bearing phosphonamidate, phosphonate, and phosphinate moieties were synthesized. Prepared compounds were evaluated in an enzymatic CDK9/CycT1 assay. The most potent molecules were tested in cell-based toxicity and HIV proliferation assays. Selectivity of shortlisted compounds against CDKs and other kinases was tested. The best compound was shown to be a highly specific, ATP-competitive inhibitor of CDK9/CycT1 with antiviral activity.