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2.
Fa Yi Xue Za Zhi ; 35(4): 396-401, 2019 Aug.
Artigo em Inglês, Chinês | MEDLINE | ID: mdl-31532145

RESUMO

Abstract: Objective To study the protein expression of cluster of differentiation 63 (CD63) in lung tissues of guinea pigs that died of anaphylactic shock and discuss the diagnostic value of CD63 for death from anaphylactic shock. Methods Twenty guinea pigs were randomly divided into control group, anaphylactic shock immediate death group, cold storage group (4 ℃ for 48 h) and frozen group (-20 ℃ for 7 d). The animal model of guinea pigs that died of anaphylactic shock was established with human mixed serum injection. The expression changes of CD63 protein and CD63 mRNA in lung tissues were detected by hematoxylin-eosin (HE) staining, immunohistochemical staining, Western blotting, enzyme-linked immunosorbent assay (ELISA) and real-time RT-PCR. Results HE staining results showed congestion, and edema of lung tissues, and eosinophil infiltration in the anaphylactic shock groups. Western blotting analysis results showed that the expression of CD63 protein in the lung tissues of guinea pigs that died of anaphylactic shock was significantly higher than that in the control group (P<0.05). Comparison between the anaphylactic shock groups was made, and the differences had no statistical significance. The results of immunohistochemical staining and real-time RT-PCR were consistent with that of Western blotting. ELISA results showed that CD63 protein expression in the immediate death group was higher than that in the control group (P<0.05). Conclusion The expression of CD63 protein and CD63 mRNA in the lung tissues of guinea pigs that died of anaphylactic shock is significantly enhanced. Animal carcasses which were put in cold storage for 48 h and frozen for 7 d do not affect the examination of the above indicators. CD63 protein is expected to become an auxiliary diagnostic indicator of death from anaphylactic shock.


Assuntos
Anafilaxia/metabolismo , Pulmão/metabolismo , Tetraspanina 30/metabolismo , Anafilaxia/mortalidade , Animais , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Cobaias , Humanos , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Soro
3.
Artigo em Japonês | MEDLINE | ID: mdl-31434811

RESUMO

Recently, the main air pollutant has been fine particulate matter (PM2.5), which is taken up by the whole body with severe adverse health effects. The main chemical components of PM2.5 are salts of sulfate (and nitrate) and carbons. However, it remains unknown which components are toxic. Here, the author reviewed the literatures to determine which components are toxic and the main mechanisms underlying their toxicity. Many epidemiological studies have shown that sulfate concentration is strongly related to mortality. However, there is no experimental evidence showing that sulfate at environmental concentrations of PM2.5 causes cardiovascular disease or other disease. On the other hand, carbon components such as elementary carbon (EC) produces high concentrations of reactive oxygen species (ROS) via its phagocytosis by macrophages, and organic carbon (OC) also produces high concentrations of ROS during its metabolic processes, and the ROS cause acute and chronic inflammation. They cause many diseases including cardiovascular disease, asthma and cancer. Furthermore, there are many lines of evidence showing that epigenetic changes such as DNA methylation or microRNA expression induced by particulate matters also induce the development of many diseases such as those mentioned above. It has been reported that carbon components are incorporated into the brain and produce ROS, and that the ROS cause damage to brain cells and Alzheimer's disease and cognitive disorders in the elderly.From these lines of evidence, the author would like to emphasize that the main toxicity of PM2.5 is due to carbon components, and it is important to take countermeasures to decrease the concentration of carbon components in ambient air.


Assuntos
Poluentes Atmosféricos/toxicidade , Carbono/toxicidade , Material Particulado/toxicidade , Sulfatos/toxicidade , Doença de Alzheimer/etiologia , Animais , Asma/etiologia , Doenças Cardiovasculares/etiologia , Epigênese Genética , Cobaias , Humanos , Camundongos , Neoplasias/etiologia , Estresse Oxidativo , Espécies Reativas de Oxigênio/metabolismo
4.
Vet Microbiol ; 235: 86-92, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31282383

RESUMO

Although PCV2 infections generally cause mild disease in pigs, concurrent co-infections with other pathogens can damage the immune system and cause more severe diseases, collectively termed porcine circovirus associated diseases (PCVAD). Involvement of porcine parvovirus (PPV, a common cause of reproductive failure in naïve dams) in PCVAD caused by PCV2, has been reported. As this co-infection can be difficult to eliminate, there is a critical need to develop an effective vaccine to protect against PPV or synergistic effects of PCV2 and PPV under field conditions. In this study, we designed chimeric PCV2 virus-like particles (cVLPs) displaying a B-cell epitope derived from PPV1 structural protein around the surface of the 2-fold axes of PCV2 VLPs, based on 3D-structure analysis of the PCV2 capsid. The cVLPs were successfully prepared, verified by transmission electron microscopy and chromatography, with robust antibody titers against PCV2 and PPV1 produced in mice and guinea pigs. In addition, in guinea pigs challenged with 106 TCID50 PCV2, cVLPs conferred more effective immune protection (based on viral load) than a commercial PCV2 vaccine. Finally, antibody responses and immune protection against PPV were also evaluated. In guinea pigs vaccinated with cVLPs, although PPV antibodies detected by a hemagglutination inhibition (HI) assay appeared later after vaccination in the PCV2 cVLPs group than in the commercial PPV vaccine group, there were fewer PPV genomic DNA copies in the PCV2 cVLPs group than in a PBS group. In conclusion, guinea pigs vaccinated with cVLPs developed effective protective immunity against PCV2 challenge, with some protective immunity against PPV. This study provided valuable research data to pursue molecular design of chimeric epitopes PCV2 VLPs.


Assuntos
Infecções por Circoviridae/veterinária , Coinfecção/veterinária , Epitopos de Linfócito B/imunologia , Imunidade Humoral , Infecções por Parvoviridae/veterinária , Vacinas Virais/imunologia , Animais , Anticorpos Neutralizantes/sangue , Anticorpos Antivirais/sangue , Linfócitos B/imunologia , Infecções por Circoviridae/imunologia , Infecções por Circoviridae/prevenção & controle , Circovirus/imunologia , Coinfecção/virologia , Feminino , Cobaias , Camundongos , Infecções por Parvoviridae/imunologia , Infecções por Parvoviridae/prevenção & controle , Parvovirus Suíno/imunologia , Suínos , Doenças dos Suínos/prevenção & controle , Doenças dos Suínos/virologia , Vacinas Atenuadas/imunologia , Vacinas de Partículas Semelhantes a Vírus/imunologia
5.
Biomed Khim ; 65(3): 227-230, 2019 Apr.
Artigo em Russo | MEDLINE | ID: mdl-31258146

RESUMO

The experimental study in vivo was aimed at evaluation of hypolipidemic action of the original natural microbial enzyme preparation of cholesterol oxidase (CHO). In preliminary chronic experiments in rats, rabbits, dogs, low toxicity, good tolerability, and anti-atherosclerotic activity of the CHO preparation were established. To assess the effect of CHO under conditions of moderate, nutritional, atherogenic dyslipoproteinemia, experiments were carried out in rats, guinea pigs, and rabbits. It was shown that administration of CHO had the pronounced lipid-lowering effect in models of atherogenic dyslipoproteinemia induced in these animals.


Assuntos
Aterosclerose/tratamento farmacológico , Colesterol Oxidase/farmacologia , Hipolipemiantes/farmacologia , Lipídeos/sangue , Animais , Cães , Cobaias , Coelhos , Ratos
6.
J Med Microbiol ; 68(9): 1314-1319, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31274404

RESUMO

Introduction. Current intradermal tuberculin skin tests for latent tuberculosis infection (LTBI) based on purified protein derivative (PPD) have poor specificity.Aims. Developing a better skin test antigen as well as a simple skin patch test may improve and facilitate diagnostic performance.Methodology. Defined recombinant antigens that were unique to Mycobacterium tuberculosis (MTB), including two potential latency-associated antigens (ESAT-6 and Rv2653c) and five DosR-encoded latency proteins (Rv1996, Rv2031c, Rv2032, DevR and Rv3716c), were used as diagnostic skin test reagents in comparison with a standard PPD. The performance of the skin tests based on the detection of delayed-type hypersensitivity (DTH) reaction in guinea pigs sensitized to MTB and M. bovis bacille Calmette-Guérin (BCG) vaccine was evaluated.Results. The latency antigens Rv1996, Rv2031c, Rv2032 and Rv2653c and the ESAT-6 protein elicited less reactive DTH skin responses in MTB-sensitized guinea pigs than those resulting from PPD, but elicited no response in BCG-vaccinated guinea pigs. The remaining two latency antigens (DevR and Rv3716c) elicited DTH responses in both groups of animals, as did PPD. The reactivity of PPD in BCG-vaccinated guinea pigs was greater than that of any of the selected skin test reagents. Using stronger concentrations of selected skin test reagents in the patch test led to increased DTH responses that were comparable to those elicited by PPD in guinea pigs sensitized with MTB.Conclusion. Transdermal application of defined purified antigens might be a promising method for LTBI screening.


Assuntos
Hipersensibilidade Tardia/imunologia , Tuberculose Latente/diagnóstico , Mycobacterium tuberculosis/imunologia , Testes Cutâneos/métodos , Adesivo Transdérmico , Animais , Antígenos de Bactérias/imunologia , Vacina BCG/imunologia , Feminino , Cobaias , Indicadores e Reagentes , Mycobacterium tuberculosis/isolamento & purificação , Testes Cutâneos/normas , Tuberculina/imunologia
7.
Acta Vet Scand ; 61(1): 27, 2019 Jun 13.
Artigo em Inglês | MEDLINE | ID: mdl-31196125

RESUMO

The aim of this study was to describe a core needle biopsy technique in the guinea pig (Cavia porcellus) and to assess the incidence of complications when applying this method. Biopsies were taken from the right hepatic lobe of 36 healthy guinea pigs under ultrasound guidance using a Tru-Cut needle. There were no immediate complications in 35 animals but ultrasound images showed a haemorrhage from the biopsy site in one guinea pig. The haemorrhage stopped after administering a sterile cooling dressing. One guinea pig died 13 days after the biopsy due to late complications. The procedure is in some animals associated with severe, potential life-threatening, complications. Assessment of the biopsy site by ultrasonography for 30 min after the procedure is recommended to allow timely handling of haemorrhage. The procedure is not recommended in animals with a suspected coagulopathy. Due to the risk of severe complications, this procedure should be restricted to guinea pigs where the result of the biopsy examination is expected to be valuable for the choice of treatment or prognosis. Owners should be made aware of the risks associated with the procedure.


Assuntos
Biópsia com Agulha de Grande Calibre/veterinária , Cobaias , Fígado/cirurgia , Medicina Veterinária/métodos , Animais , Biópsia com Agulha de Grande Calibre/efeitos adversos , Biópsia com Agulha de Grande Calibre/normas , Hemorragia/etiologia , Hemorragia/veterinária , Ultrassonografia
8.
Vestn Otorinolaringol ; 84(2): 4-7, 2019.
Artigo em Russo | MEDLINE | ID: mdl-31198207

RESUMO

AIM: To study ultrastructural changes in stria's vascularis cells of inner ear and determine possible ways of correction. MATERIAL AND METHODS: The work was carried out on male guinea pigs. After completion of the experiment, stria vascularis of inner ear was subjected to electron microscopic examination. RESULTS AND DISCUSSION: In the control group (receiving gentamycin sulfate in an ototoxic dose), signs of blood flow disturbance were revealed, as well as ultrastructural changes in stria's vascularis cells (expansion of intercellular space, deformation of organelles, thinning of glycocalyx, blebbing). Also, fragmented cells were found. These changes are characteristics for apoptosis. In experimental group (receiving gentamycin sulfate and melaxen), degenerative changes were less pronounced. An increase of cell's secretory activity was observed. CONCLUSION: Changes in stria's vascularis cells by using melaxen are less pronounced. Increase of cell's secretory activity in stria vascularis is a compensatory reaction and saves auditory function.


Assuntos
Surdez , Orelha Interna , Perda Auditiva , Animais , Cóclea , Orelha Interna/fisiopatologia , Cobaias , Audição , Masculino , Estria Vascular
9.
Int J Nanomedicine ; 14: 4211-4227, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31239676

RESUMO

Background: Cisplatin is an extensively used anti-neoplastic agent for the treatment of various solid tumors. However, a high incidence of severe ototoxicity is accompanied by its use in the clinic. Currently, no drugs or therapeutic strategies have been approved for the treatment of cisplatin-induced ototoxicity by the FDA. Purpose: The purpose of this study was to investigate the otoprotective effects of dexamethasone (DEX)-loaded silk-polyethylene hydrogel (DEX-SILK) following round window membrane administration in the cisplatin-induced ototoxicity mouse model. Methods: The morphology, gelation kinetics, viscosity and secondary structure of the DEX-SILK hydrogel were analyzed. DEX concentration in the perilymph was tested at different time points following hydrogel injection on the RWM niche. Cultured cells (HEI-OC1), organ of Corti explants (C57/BL6, P0-2), and cisplatin-induced hearing loss mice model (C57/BL6) were used as in vitro and in vivo models for investigating the otoprotective effects of DEX-SILK hydrogel against cisplatin. Results: Encapsulation of DEX with a loading of 8% (w/v) did not significantly change the silk gelation time, and DEX was evenly distributed in the Silk-PEG hydrogel as visualized by scanning electron microscopy (SEM). The concentration of Silk majorly influenced DEX distribution, morphological characteristics, viscosity, and gelation time. The optimized DEX-SILK hydrogel (8% w/v loading, 15% silk concentration, 10 µl) was administered directly onto the RWM of the guinea pigs. The DEX concentration in the perilymph was maintained above 1 µg/ml for at least 21 days for the DEX-SILK, while it was maintained for less than 6 h in the control sample of free DEX. DEX-SILK (5-60 ng/ml) exhibited significant protective effects against cisplatin-induced cellular ototoxicity and notably reduced the production of reactive oxygen species (ROS). Eventually, pretreatment with DEX-SILK effectively preserved outer hair cells in the cultured organ of Corti explants and demonstrated significant hearing protection at 4, 8, and 16 kHz in the cisplatin-induced hearing loss mice as compared to the effects noted following pretreatment with DEX. Conclusion: These results demonstrated the clinical value of DEX-SILK for the therapy of cisplatin-induced ototoxicity.


Assuntos
Cisplatino/efeitos adversos , Dexametasona/farmacologia , Orelha/patologia , Hidrogéis/química , Injeções , Polietilenoglicóis/química , Seda/química , Animais , Materiais Biocompatíveis/efeitos adversos , Bombyx , Linhagem Celular , Cóclea/efeitos dos fármacos , Cóclea/patologia , Dexametasona/uso terapêutico , Modelos Animais de Doenças , Liberação Controlada de Fármacos , Cobaias , Perda Auditiva/induzido quimicamente , Perda Auditiva/tratamento farmacológico , Camundongos Endogâmicos C57BL , Substâncias Protetoras/farmacologia , Espécies Reativas de Oxigênio/metabolismo
10.
Int J Nanomedicine ; 14: 3679-3689, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31239660

RESUMO

Background and purpose: Conventional topical ophthalmic aqueous solutions and suspensions are often associated with low bioavailability and high administration frequency, pulsatile dose and poor exposure to certain ocular parts. The aim of this study was to develop an ophthalmic nanoparticles loaded gel, for delivering prednisolone acetate (PA), to increase dosing accuracy, bioavailability, and accordingly, efficiency of PA in treating inflammatory ocular diseases. Methods: A novel formulation of self-assembled nanoparticles was prepared by the complexation of chitosan (CS) and, the counter-ion, sodium deoxycholate (SD), loaded with the poorly-water-soluble PA. Particle size, zeta potential, encapsulation efficiency (EE) and drug loading content (LC) of prepared nanoparticles were assessed. Moreover, the nanoparticles were characterized using differential scanning calorimetry (DSC) and Fourier transform infrared spectroscopy (FTIR). Drug release and eye anti-inflammatory potential of the prepared novel formulation was investigated. Results: Mean particle size of the nanoparticles have dropped from 976 nm ±43 (PDI 1.285) to 480 nm ±28 (PDI 1.396) when the ratio of CS-SD was decreased. The incorporation of 0.1-0.3% of polyvinyl alcohol (PVA), in the preparation stages, resulted in smaller nanoparticles: 462 nm ±19 (PDI 0.942) and 321 nm ±22 (PDI 0.454) respectively. DSC and FTIR results demonstrated the interaction between CS and SD, however, no interactions were detected between PA and CS or SD. Drug release of PA as received, in simulated tears fluid (pH 7.4), showed a twofold increase (reaching an average of 98.6% in 24 hours) when incorporated into an optimized nanoparticle gel formulation (1:5 CS-SD). Conclusion: The anti-inflammatory effect of PA nanoparticles loaded gel on female guinea pig eyes was significantly superior to that of the micronized drug loaded gel (P < 0.05).


Assuntos
Anti-Inflamatórios/farmacologia , Quitosana/química , Ácido Desoxicólico/química , Olho/efeitos dos fármacos , Olho/patologia , Nanopartículas/química , Prednisolona/análogos & derivados , Animais , Varredura Diferencial de Calorimetria , Portadores de Fármacos/química , Liberação Controlada de Fármacos , Feminino , Cobaias , Inflamação/patologia , Nanopartículas/ultraestrutura , Tamanho da Partícula , Prednisolona/farmacologia , Espectroscopia de Infravermelho com Transformada de Fourier
11.
Bioelectrochemistry ; 129: 170-178, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31181439

RESUMO

Our aim was to investigate if the cardioplegic solution HTK can be improved by the addition of the ROS scavenger melatonin. 158 guinea pig hearts without (UI80) or with HTK protection (HTK80) were investigated in ischemia/reperfusion experiments. Ischemia lasted 80 min at 30 °C. Melatonin was given before ischemia (UI80 + M1, HTK80 + M1) or before and after ischemia (UI80 + M2, HTK80 + M2). We measured the left ventricular developed pressure (LVDP), diastolic pressure (LVPmin), cardiac rhythm (VC-RR), time of electrical cell uncoupling (t-in) and recovery (t-ret), intracellular Ca++ [Ca++], and postischemic ROS. After 45 min reperfusion, LVDP in UI80 was significantly higher than in HTK80 (p < .01). Compared to UI80, the postischemic ROS burst was slightly smaller in HTK80 and significantly smaller in HTK80 + M1 and HTK80 + M2 (p < .05). Melatonin had no effect on LVPmin, t-in, t-ret, [Ca++], and on LVDP in groups UI80 + M1 and HTK80 + M1, improved slightly VC-RR (n. s.) but significantly decreased LVDP in the groups UI80 + M2 and HTK80 + M2 (p < .01). With melatonin we were able to attenuate the postischemic ROS burst, but the tissue damage by ROS seemed to be less important for the chosen ischemia condition because melatonin was unable to improve the functional recovery during reperfusion of HTK protected hearts.


Assuntos
Soluções Cardioplégicas/uso terapêutico , Depuradores de Radicais Livres/uso terapêutico , Parada Cardíaca Induzida/métodos , Coração/efeitos dos fármacos , Melatonina/uso terapêutico , Traumatismo por Reperfusão Miocárdica/terapia , Animais , Feminino , Cobaias , Coração/fisiopatologia , Frequência Cardíaca/efeitos dos fármacos , Traumatismo por Reperfusão Miocárdica/tratamento farmacológico , Traumatismo por Reperfusão Miocárdica/fisiopatologia , Traumatismo por Reperfusão Miocárdica/cirurgia
12.
Adv Exp Med Biol ; 1124: 217-231, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31183829

RESUMO

Seminal vesicles (SVs), a pair of male accessory glands, contract upon sympathetic nerve excitation during ejaculation while developing spontaneous phasic constrictions in the inter-ejaculatory storage phase. Recently, the fundamental role of the mucosa in generating spontaneous activity in SV of the guinea pig has been revealed. Stretching the mucosa-intact but not mucosa-denuded SV smooth muscle evokes spontaneous phasic contractions arising from action potential firing triggered by electrical slow waves and associated Ca2+ flashes. These spontaneous events primarily depend on sarco-endoplasmic reticulum (SR/ER) Ca2+ handling linked with the opening of Ca2+-activated chloride channels (CaCCs) resulting in the generation of slow waves. Slow waves in mucosa-intact SV smooth muscle are abolished upon blockade of gap junctions, suggesting that seminal smooth muscle cells are driven by cells distributed in the mucosa. In the SV mucosal preparations dissected free from the smooth muscle layer, a population of cells located just beneath the epithelium develop spontaneous Ca2+ transients relying on SR/ER Ca2+ handling. In the lamina propria of the SV mucosa, vimentin-immunoreactive interstitial cells including platelet-derived growth factor receptor α (PDGFRα)-immunoreactive cells are distributed, while known pacemaker cells in other smooth muscle tissues, e.g. c-Kit-positive interstitial cells or α-smooth muscle actin-positive atypical smooth muscle cells, are absent. The spontaneously-active subepithelial cells appear to drive spontaneous activity in SV smooth muscle either by sending depolarizing signals or by releasing humoral substances. Interstitial cells in the lamina propria may act as intermediaries of signal transmission from the subepithelial cells to the smooth muscle cells.


Assuntos
Células Intersticiais de Cajal/fisiologia , Membrana Mucosa/fisiologia , Contração Muscular , Músculo Liso/fisiologia , Glândulas Seminais/fisiologia , Animais , Sinalização do Cálcio , Cobaias , Canais Iônicos/fisiologia , Masculino , Miócitos de Músculo Liso/fisiologia
13.
Nihon Yakurigaku Zasshi ; 153(6): 273-277, 2019.
Artigo em Japonês | MEDLINE | ID: mdl-31178532

RESUMO

Continuous and real-time measurement of local concentrations of systemically administered drugs in vivo must be crucial for pharmacological studies. Nevertheless, conventional methods require considerable samples quantity and have poor sampling rates. Additionally, they cannot determine how drug kinetics correlates with target function over time. Here, we describe a system with two different sensors. One is a needle-type microsensor composed of boron-doped diamond with a tip of ~40 µm in diameter, and the other is a glass microelectrode. We first tested bumetanide. This diuretic can induce deafness. In the guinea-pig cochlea injected intravenously with bumetanide, the changes of the drug concentration and the extracellular potential underlying hearing were simultaneously measured in real time. We further examined an antiepileptic drug lamotrigine in the rat brain, and tracked its kinetics and at the same time the local field potentials representing neuronal activity. The action of the anticancer reagent doxorubicin was also monitored in the cochlea. This microsensing system may be applied to analyze pharmacokinetics and pharmacodynamics of various drugs at local sites in vivo, and contribute to promoting the pharmacological researches.


Assuntos
Boro , Cóclea/efeitos dos fármacos , Diamante , Doxorrubicina/farmacologia , Microeletrodos , Animais , Encéfalo/efeitos dos fármacos , Bumetanida/farmacologia , Cobaias , Lamotrigina/farmacologia , Neurônios/efeitos dos fármacos , Ratos
14.
Klin Lab Diagn ; 64(5): 308-313, 2019.
Artigo em Russo | MEDLINE | ID: mdl-31185155

RESUMO

The most common pathology in the clinic of orthopedic dentistry is the presence of partial adentia in patients, manifested in the form of defects of dentition of various localization and length. Removable orthopedic structures in the oral cavity are a potential place for adhesion and colonization of microorganisms. The aim of the research was to study Candida albicans biofilms on the surface of base plastics of removable orthopedic structures using scanning electron microscopy. 175 cultures of C. albicans were isolated and identified from the oral mucosa of patients at various stages of orthopedic rehabilitation. When studying the surface of samples of plastics of hot and cold type polymerization and Candida biofilms using a JEOL JCM 5700 scanning electron microscope (JEOL, Japan), features of biofilm formation were established. An assessment of the nature of the manifestation of the hemagglutinating activity of clinical strains of Candida fungi in the hemagglutination test with human erythrocytes I (O), II (A) of the human and guinea pig blood groups was carried out. The total number of hemagglutinating strains was 37.14%, with the prevalence of the proportion of manna-resistant (MRHA) cultures - 23.43% of cases. Micrographs of the C. albicans yeast-like biofilm biofilm were obtained on the surface of hot and cold-type plastics in incubation dynamics. Scanning electron microscopy revealed the most pronounced changes in the surface of hot plastics of polymerization compared to cold plastics with long incubation of C. albicans, which characterize the loosening of plastics and the appearance of cracks on the surface, and the cracking of a yeast-like fungus biofilm was noted.


Assuntos
Biofilmes , Candida albicans/ultraestrutura , Implantes Dentários/microbiologia , Microscopia Eletrônica de Varredura , Equipamentos Ortopédicos/microbiologia , Plásticos , Animais , Candida albicans/crescimento & desenvolvimento , Contaminação de Equipamentos , Cobaias , Humanos , Japão
15.
Forensic Sci Int ; 301: 341-349, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31202147

RESUMO

A rapid, sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination and quantification of 12 psychotropic drugs and metabolites in hair was developed and validated. After freeze grinding with methanol, the supernatant was determined by LC-MS/MS using an Allure PFPPropyl column (100 × 2.1 mm, 5 µm) with a gradient elution of acetonitrile and 10 mmol/L ammonium acetate with 0.1% formic acid, and in the subsequent analysis using multiple reaction monitoring (MRM) mode, two ion transitions were monitored for each analyte. The limits of detection ranged from 0.002 to 0.05 ng/mg, and the limits of quantitation were in the range of 0.005-0.1 ng/mg. Good linearity (r > 0.995) was observed for all analytes over the linear range. Acceptable intraday and interday precision (RSD ≤ 20%) and accuracy (85.3%-112.9%) were achieved. This method of detection was applied to the analysis of guinea pig hair roots after a single dose of quetiapine. Quetiapine and 7-hydroxyquetiapine were both detected in guinea pig hair roots from 5 min post administration. The concentration of quetiapine (10.3-1733.8 ng/mg) was much higher than that of 7-hydroxyquetiapine (0.1-40.6 ng/mg) in the hair roots of guinea pigs, and higher concentrations of quetiapine and 7-hydroxyquetiapine occurred in black hair root than in that of white and brown. The animal experiment demonstrated that hair roots may be a good specimen for proving acute quetiapine poisoning when other biological matrices are not available.


Assuntos
Cabelo/química , Psicotrópicos/análise , Animais , Cromatografia Líquida , Dibenzotiazepinas/análise , Toxicologia Forense , Cobaias , Limite de Detecção , Modelos Animais , Envenenamento/diagnóstico , Fumarato de Quetiapina/análise , Detecção do Abuso de Substâncias/métodos , Espectrometria de Massas em Tandem
16.
Int. j. morphol ; 37(2): 416-422, June 2019. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1002237

RESUMO

To contribute knowledge of an important experimental model for studies on skin embryology, a study was carried out to describe the morphological events of the skin during the intrauterine life of Cavia porcellus from the 10th to the 60th day of gestation. Embryos and fetuses were dissected, and the skin of the nasal, cranial, lumbar and anal regions was processed byoptical microscopy. At 30 days the first hairs, called lanugos, were observed in the cranial region. The morphological description showed that a few days can make a great difference in development.


Con el objetivo de contribuir al conocimiento de un importante modelo experimental para estudios sobre embriología de la piel, se llevó a cabo un estudio para describir los eventos morfológicos de la piel durante la vida intrauterina de Cavia porcellus desde el día 10 hasta el día 60 de gestación. Los embriones y los fetos se disecaron y se procesó la piel de las regiones nasal, craneal, lumbar y anal, mediante microscopía óptica. A los 30 días se observaron los primeros vellos, llamados lanugos, en la región craneal. La descripción morfológica mostró que unos pocos días pueden marcar una gran diferencia en el desarrollo.


Assuntos
Animais , Masculino , Gravidez , Pele/embriologia , Modelos Animais , Cobaias , Pele/crescimento & desenvolvimento , Pele/ultraestrutura , Fatores de Tempo
17.
Vet Microbiol ; 233: 113-117, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31176396

RESUMO

Bovine ephemeral fever virus (BEFV) causes an acute febrile disease in cattle and water buffalo. The disease has an impact on dairy and beef production in tropical and subtropical countries. Vaccination is used for disease prevention and control. In this study, we developed a recombinant lentivirus to produce mammalian stable cells expressing histidine-tagged BEFV G protein with a deleted transmembrane domain (GΔTM) as a secretory protein. In addition, guinea pigs were immunised with the purified GΔTM protein and booster immunised at a 3-week interval. The mammalian stable cells were able to continuously produce GΔTM protein for a minimum of 25 passages. All of the mammalian stable cells expressing GΔTM protein could react specifically with a BEFV convalescent bovine serum. Serum samples from the immunised guinea pigs could react strongly and specifically with the purified GΔTM protein. Moreover, post-immunised guinea pig sera contained antibodies that could neutralise BEFV. These results indicate that the G protein without a transmembrane domain can be used as a subunit vaccine for the prevention and control of BEFV. The availability of the mammalian stable cells, which constitutively express GΔTM protein, could facilitate the potential use of the secretory protein for BEFV diagnosis and vaccine development.


Assuntos
Anticorpos Antivirais/sangue , Febre Efêmera/prevenção & controle , Glicoproteínas/genética , Glicoproteínas/imunologia , Imunogenicidade da Vacina , Proteínas Virais/genética , Proteínas Virais/imunologia , Animais , Anticorpos Neutralizantes/sangue , Bovinos , Linhagem Celular , Febre Efêmera/imunologia , Vírus da Febre Efêmera Bovina , Feminino , Cobaias , Células HEK293 , Humanos , Transfecção , Vacinação , Vacinas Virais/imunologia
18.
BMC Bioinformatics ; 20(Suppl 10): 250, 2019 May 29.
Artigo em Inglês | MEDLINE | ID: mdl-31138104

RESUMO

BACKGROUND: Drug candidates often cause an unwanted blockage of the potassium ion channel of the human ether-a-go-go-related gene (hERG). The blockage leads to long QT syndrome (LQTS), which is a severe life-threatening cardiac side effect. Therefore, a virtual screening method to predict drug-induced hERG-related cardiotoxicity could facilitate drug discovery by filtering out toxic drug candidates. RESULT: In this study, we generated a reliable hERG-related cardiotoxicity dataset composed of 2130 compounds, which were carried out under constant conditions. Based on our dataset, we developed a computational hERG-related cardiotoxicity prediction model. The neural network model achieved an area under the receiver operating characteristic curve (AUC) of 0.764, with an accuracy of 90.1%, a Matthews correlation coefficient (MCC) of 0.368, a sensitivity of 0.321, and a specificity of 0.967, when ten-fold cross-validation was performed. The model was further evaluated using ten drug compounds tested on guinea pigs and showed an accuracy of 80.0%, an MCC of 0.655, a sensitivity of 0.600, and a specificity of 1.000, which were better than the performances of existing hERG-toxicity prediction models. CONCLUSION: The neural network model can predict hERG-related cardiotoxicity of chemical compounds with a high accuracy. Therefore, the model can be applied to virtual high-throughput screening for drug candidates that do not cause cardiotoxicity. The prediction tool is available as a web-tool at http://ssbio.cau.ac.kr/CardPred .


Assuntos
Cardiotoxicidade/metabolismo , Canais de Potássio Éter-A-Go-Go/metabolismo , Redes Neurais (Computação) , Animais , Área Sob a Curva , Bases de Dados Genéticas , Canais de Potássio Éter-A-Go-Go/química , Cobaias , Humanos , Aprendizado de Máquina , Curva ROC
19.
Adv Mater ; 31(27): e1901103, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31112631

RESUMO

Ionic liquids (ILs) and deep eutectic solvents have shown great promise in drug delivery applications. Choline-based ILs, in particular choline and geranic acid (CAGE), have been used to enhance the transdermal delivery of several small and large molecules. However, detailed studies outlining the design principles of ILs for transdermal drug delivery are still lacking. Using two model drugs of differing hydrophilicities, acarbose and ruxolitinib and 16 ILs, the dependence of skin penetration on the chemical properties of ILs is examined. First, the impact of ion stoichiometry on skin penetration of drugs is assessed using CAGE, which evidences that a molar ratio of 1:2 of choline to geranic acid yields the highest delivery. Subsequently, variants of CAGE are prepared using anions with structural similarity to geranic acid and cations with structural similarity to choline at a ratio of 1:2. Mechanistic studies reveal that the potency of ILs in enhancing transdermal drug delivery correlates inversely with the inter-ionic interactions as determined by 2D NMR spectroscopy. Using this understanding, a new IL is designed, and it provides the highest delivery of ruxolitinib of all ILs tested here. Overall, these studies provide a generalized framework for optimizing ILs for enhancing skin permeability.


Assuntos
Acarbose/administração & dosagem , Ácidos Carboxílicos/química , Portadores de Fármacos/química , Líquidos Iônicos/química , Pirazóis/administração & dosagem , Administração Cutânea , Animais , Colina/química , Cobaias , Concentração de Íons de Hidrogênio , Interações Hidrofóbicas e Hidrofílicas , Permeabilidade , Pele/metabolismo , Solventes/química , Terpenos/química
20.
BMC Ophthalmol ; 19(1): 102, 2019 May 03.
Artigo em Inglês | MEDLINE | ID: mdl-31053109

RESUMO

BACKGROUND: Lens-induced myopization in guinea pigs has been used as model for the process of myopization in humans. It has not been explored yet whether the change in globe shape in eyes undergoing myopization is similar in experimental myopia in guinea pigs and in clinical myopia in patients. METHODS: The study included 70 guinea pigs (age:2-3 weeks) equally divided into a study group with lens-induced myopization for 5 weeks, and a control group wearing goggles with no refractive power. The globe diameters were measured using a microcaliper after enucleation. RESULTS: The horizontal globe diameter (9.19 ± 0.15 mm versus 9.15 ± 0.18 mm; P = 0.25) and vertical globe diameter (9.02 ± 0.11 mm versus 8.99 ± 0.14 mm; P = 0.29) did not differ significantly between the study group and control group. The sagittal diameter was significantly longer in the study group (8.96 ± 0.15 mm versus 8.84 ± 0.14 mm; P = 0.001). While the vertical and horizontal globe diameters were correlated with each other in a ratio of approximately 1:1 (non-standardized regression coefficient B:0.94;95% confidence interval (CI):0.73,1.15), the steepness of the regression lines of the associations of both diameters with the sagittal diameter were flatter (horizontal to sagittal diameter: B: 0.64; 95% CI: 0.44,0.83; vertical to sagittal diameter:B:0.55;95% CI:0.41,0.69). Correspondingly, the ratios of horizontal-to-sagittal globe diameter and of vertical-to-sagittal globe diameter decreased (P < 0.001) with longer sagittal diameter. CONCLUSIONS: For each mm axial elongation in young guinea pigs the horizontal globe diameter increased by 0.64 mm (95%CI:0.44,0.83) and the vertical diameter by 0.55 mm (95% CI:0.41,0.69), indicating that the globe enlargement occurred predominantly in the sagittal direction. Axial elongation in guinea pigs led to a similar relative change in ocular shape as in humans.


Assuntos
Comprimento Axial do Olho/fisiologia , Miopia/fisiopatologia , Animais , Modelos Animais de Doenças , Cobaias , Análise de Regressão
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