RESUMEN
Transcriptomic and proteomic analysis were performed on 72 h biofilms of the acneic strain Cutibacterium acnes and planktonic cultures in the presence of epinephrine. Epinephrine predominantly downregulated genes associated with various transporter proteins. No correlation was found between proteomic and transcriptomic profiles. In control samples, the expression of 51 proteins differed between planktonic cultures and biofilms. Addition of 5 nM epinephrine reduced this number, and in the presence of 5 µM epinephrine, the difference in proteomic profiles between planktonic cultures and biofilms disappeared. According to the proteomic profiling, epinephrine itself was more effective in the case of C. acnes biofilms and potentially affected the tricarboxylic acid cycle (as well as alpha-ketoglutarate decarboxylase Kgd), biotin synthesis, cell division, and transport of different compounds in C. acnes cells. These findings are consistent with recent research on Micrococcus luteus, suggesting that the effects of epinephrine on actinobacteria may be universal.
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In this study, the effect of epinephrine on the biofilm formation of Micrococcus luteus C01 isolated from human skin was investigated in depth for the first time. This hormone has a complex effect on biofilms in various systems. In a system with polytetrafluoroethylene (PTFE) cubes, treatment with epinephrine at a physiological concentration of 4.9 × 10-9 M increased the total amount of 72-h biofilm biomass stained with crystal violet and increased the metabolic activity of biofilms, but at higher and lower concentrations, the treatment had no significant effect. On glass fiber filters, treatment with the hormone decreased the number of colony forming units (CFUs) and changed the aggregation but did not affect the metabolic activity of biofilm cells. In glass bottom plates examined by confocal microscopy, epinephrine notably inhibited the growth of biofilms. RNA-seq analysis and RT-PCR demonstrated reproducible upregulation of genes encoding Fe-S cluster assembly factors and cyanide detoxification sulfurtransferase, whereas genes encoding the co-chaperone GroES, the LysE superfamily of lysine exporters, short-chain alcohol dehydrogenase and the potential c-di-GMP phosphotransferase were downregulated. Our results suggest that epinephrine may stimulate matrix synthesis in M. luteus biofilms, thereby increasing the activity of NAD(H) oxidoreductases. Potential c-di-GMP pathway proteins are essential in these processes.
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The article presents the results of spectrophotometric tooth enamel scanning for differential diagnosis of focal enamel demineralization and fluorosis. Research was conducted in vivo on teeth affected by these diseases. VITA EasyShade spectrophotometer measurements were made on the affected area and on the visually healthy part of enamel. The lightness appeared as the only one differential significant optical characteristics of tooth enamel. Lightness metrics were higher in the case of initial caries than on the healthy part of enamel when these metrics were lower in the case of fluorosis than on the healthy part of enamel.
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Fluorosis Dental , Espectrofotometría , Desmineralización Dental , Caries Dental , Esmalte Dental , Diagnóstico Diferencial , Fluorosis Dental/diagnóstico , Humanos , Desmineralización Dental/diagnósticoRESUMEN
The article presents the results of spectrophotometric tooth enamel scanning. Research was conducted in vitro on extracted teeth. Numeric color parameters measurements were made by VITA EasyShade spectrophotometer after the 37% phosphoric acid enamel conditioning, that was made for an artificial tooth enamel demineralization reconstruction. Then transverse section of this enamel part was evaluated by scanning electronic microscopy. The scale of enamel demineralization depth in mm/microns and its' lightness parameter compliance was created.
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Esmalte Dental/patología , Desmineralización Dental/diagnóstico , Humanos , Microscopía Electrónica de Rastreo , Espectrofotometría/métodos , Desmineralización Dental/inducido químicamente , Desmineralización Dental/patologíaRESUMEN
The article describes focal enamel demineralization - the most common disease of teeth hard tissues. Local and general factors of this pathological process advent and development were described. The main tools of focal enamel demineralization diagnostics and treatment were observed. The main role of enamel mesoporous structure in the remineralization therapy possibility was underlined. There are still a lot of questions in focal enamel demineralization diagnostics after publications analysis because of the existing methods subjectivity. The question of indications for different remineralization tools applications optimization depending on the focal enamel demineralization degree is still open.
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Esmalte Dental/patología , Desmineralización Dental/diagnóstico , Desmineralización Dental/terapia , Remineralización Dental/métodos , HumanosRESUMEN
A total of 44 serum specimens from 7 patients with kidneys or liver transplanted from donors who had antibodies (Ab) to human cytomegalovirus (CMV) were studied. In 4 recipients anti-CMV Ab were found before transplantation and in 3 others they were not detected. It was shown by EIA that IgM and IgG anti-CMV appeared in the sera of primarily infected patients after 1-2 weeks and their titers were 5-10 times lower than in patients with reactivated CMV infection. Immunoblotting of Ab to individual CMV proteins showed a narrower spectrum of Ab during the initial period of primary CMV infection in comparison with the same period of reactivation and delayed production of AB to conformation-dependent determinants. Hence, analysis of anti-CMV Ab during the first 4-6 weeks after organ transplantation by EIA and immunoblotting differentiates primary CMV infection from its reactivation by Ab titers and spectrum. These parameters vary in some patients.
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Anticuerpos Antivirales/inmunología , Infecciones por Citomegalovirus/inmunología , Citomegalovirus/inmunología , Antígenos Virales/química , Antígenos Virales/inmunología , Infecciones por Citomegalovirus/etiología , Humanos , Inmunidad , Epítopos Inmunodominantes , Infecciones Oportunistas/inmunología , Trasplante de Órganos/efectos adversos , Conformación Proteica , Relación Estructura-Actividad , Inmunología del Trasplante , Trasplante HomólogoAsunto(s)
Anticuerpos Antivirales/análisis , Antígenos Virales/análisis , Infecciones por Citomegalovirus/diagnóstico , Citomegalovirus/genética , Citomegalovirus/inmunología , ADN Viral/análisis , Trasplante de Órganos , Complicaciones Posoperatorias/diagnóstico , Anticuerpos Antivirales/sangre , Anticuerpos Antivirales/orina , Antígenos Virales/sangre , Antígenos Virales/orina , Biomarcadores/análisis , Biomarcadores/sangre , Biomarcadores/orina , Citomegalovirus/patogenicidad , Infecciones por Citomegalovirus/inmunología , Infecciones por Citomegalovirus/virología , ADN Viral/sangre , ADN Viral/orina , Humanos , Técnicas para Inmunoenzimas , Inmunoglobulina G/análisis , Inmunoglobulina G/sangre , Inmunoglobulina G/inmunología , Inmunoglobulina G/orina , Inmunoglobulina M/análisis , Inmunoglobulina M/sangre , Inmunoglobulina M/inmunología , Inmunoglobulina M/orina , Reacción en Cadena de la Polimerasa , Complicaciones Posoperatorias/inmunología , Complicaciones Posoperatorias/virología , Saliva/inmunología , Saliva/virologíaRESUMEN
The organization of the mitotic apparatus was studied in human embryo lung fibroblasts (HEL) and Vero cells at 4 days postinfection with human cytomegalovirus (HCMV) strain AD 169. The bipolar spindle was detected by immunofluorescence in p72-positive mitotic cells exhibiting a regular or C-metaphase-like chromosome configuration. Electron-microscopic study of C-metaphase-like cells revealed alteration of the centrosome structure which is characterized by the following features: (1) breakdown of the diplosome, (2) separation of the fibrillar material from centrioles, and (3) disruption of the centriolar cylinder. The spindle pole in the aberrant mitotic cells consisted of one or several foci of microtubules converging on the fibrillar aggregates. There are not any signs of the nuclear envelope reconstruction found in mitotic cells with highly condensed scattered chromosomes. Unlike in HEL cells, viral particles were not detected in Vero cells. A question arises as to whether centrosome injury is an integral part of the events leading to cell death unrelated to the reproduction of HCMV.
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Ciclo Celular , Centrosoma/ultraestructura , Citomegalovirus/fisiología , Huso Acromático/ultraestructura , Anafase , Animales , Antígenos Virales/análisis , Línea Celular , Polaridad Celular , Centriolos/ultraestructura , Centriolos/virología , Centrosoma/virología , Chlorocebus aethiops , Humanos , Pulmón , Microscopía Electrónica , Microtúbulos/ultraestructura , Microtúbulos/virología , Mitosis , Índice Mitótico , Huso Acromático/virología , Factores de Tiempo , Células VeroRESUMEN
Twenty-six mouse hybridomas producing monoclonal antibodies (MAB) to human cytomegalovirus (CMV) proteins have been obtained. MAB produced by three hybridomas were studied in detail. MAB were active in indirect immunofluorescence and solid-phase enzyme immunoassay, being directed to the super-early viral protein p72. Use of the resultant MAB for analysis of CMV-infected cells demonstrated that by specificity and sensitivity of viral antigen detection they were not inferior to anti-CMV antibodies manufactured by Ortho, USA. Screening of 258 patients with suspected CMV infection showed that these MAB may be used for immunofluorescent detection of CMV antigens in the material isolated from patients and infected subjects.
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Anticuerpos Monoclonales/inmunología , Antígenos Virales/inmunología , Citomegalovirus/aislamiento & purificación , Proteínas Inmediatas-Precoces/inmunología , Animales , Anticuerpos Monoclonales/biosíntesis , Células Cultivadas , Citomegalovirus/inmunología , Infecciones por Citomegalovirus/diagnóstico , Técnica del Anticuerpo Fluorescente , Humanos , Hibridomas , Técnicas para Inmunoenzimas , Ratones , Sensibilidad y EspecificidadRESUMEN
Mutant clones of human hepatocarcinoma PLC-PRF-5 cells carrying a hepatitis B virus (HBV) genome have been obtained using selection for resistance to the toxic action of a variety of preparations to induce cell differentiation. The clones differed in various features such as expression of alpha-fetoprotein (AFP) and albumin as well as in growth rates, ability to grow in semisolid media and to be cloned in agar. Expression of the surface antigen (HBsAg) was significantly increased in mutant cells exhibiting differentiation features in contrast to the parental cells. In addition, the core antigen (HBcAg), which was silent in the original cells, was detected in some clones. There was no temporal correlation between the peak of enhanced expression of HBsAg and activation of HBcAg observed at different life periods of each clone. Evidence of cell fusion in cell culture such as premature chromatin condensation and increased numbers of binucleate cells was detected in clones with differentiation features and an increased level of viral gene expression. The approach used in this study can be used to develop cell lines of the same origin with different degrees of differentiation whilst maintaining HBV expression. This model system may be useful in the study of HBV.
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Carcinoma Hepatocelular/microbiología , Antígenos del Núcleo de la Hepatitis B/genética , Antígenos de Superficie de la Hepatitis B/genética , Virus de la Hepatitis B/genética , Neoplasias Hepáticas/microbiología , Diferenciación Celular , División Celular , Células Clonales , Resistencia a Medicamentos , Regulación Viral de la Expresión Génica , Antígenos del Núcleo de la Hepatitis B/análisis , Antígenos de Superficie de la Hepatitis B/análisis , Humanos , Modelos Genéticos , Mutagénesis , Células Tumorales Cultivadas/efectos de los fármacosRESUMEN
A test system using monoclonal antibodies to HIV p24 was developed for solid-phase ELISA for detection of HIV antigen (AG) which helped detect the content of AG in samples with trace concentrations, less than 25 pg/ml. The test system can be used for AG monitoring in natural specimens (serum of HIV-infected patients and culture medium of infected cells) and for determination of antigen-recombinant product of HIV gag gene.
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Anticuerpos Monoclonales , Antígenos VIH/sangre , Proteína p24 del Núcleo del VIH/inmunología , VIH-1/inmunología , Serodiagnóstico del SIDA/métodos , Humanos , Técnicas para Inmunoenzimas , Proteínas Recombinantes/inmunología , Sensibilidad y EspecificidadRESUMEN
The paper describes the enzyme immunoassay system for detection of human immunodeficiency virus antigens, which is based on the use of rabbit anti-HIV antibodies and monoclonal antibodies to HIV-1 gene proteins gag. The system may be useful in the examination of laboratory and clinical samples to reveal both free and conjugated antigens in the composition of immune complexes. The sensitivity of the assay system under development is 0.5 ng/ml at 100% specificity.
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Síndrome de Inmunodeficiencia Adquirida/inmunología , Anticuerpos Monoclonales/inmunología , Productos del Gen gag/inmunología , Antígenos VIH/análisis , VIH-1/inmunología , Sueros Inmunes/inmunología , Síndrome de Inmunodeficiencia Adquirida/diagnóstico , Síndrome de Inmunodeficiencia Adquirida/genética , Animales , Anticuerpos Monoclonales/genética , Antígenos VIH/inmunología , Humanos , Sueros Inmunes/genética , Técnicas para Inmunoenzimas , ConejosRESUMEN
Five strains of hybrid cells producing highly active monoclonal antibodies (MCA) to Venezuelan equine encephalomyelitis (VEE) virus were generated. VEEC6 MCA had high virus-neutralizing and hemagglutinating activities attesting to their direction to the "critical" site of E2 glycoprotein. MCA VEEA5 directed to the same glycoprotein did not overlap spatially with the previous one. MCA VEEB5 and VEEA4 interacted with conformational virus determinants taking part in hemagglutination.