Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 15 de 15
Filtrar
Mais filtros










Base de dados
Intervalo de ano de publicação
1.
Forensic Sci Med Pathol ; 15(3): 399-403, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31250258

RESUMO

Rapid identification of pathogenic agents is important in response to the emergence of biocrime and bioterrorism, to facilitate appropriate confinement and treatment. As the rapid determination system of viral genome sequences (RDV method) using exhaustive gene amplification is useful for rapid identification, we examined whether this method could be applied to forensic samples. To detect pathogenic virus in a cat with suspected viral infections, fluid swab samples were applied to the RDV method. The following steps were performed: viral propagation, extraction of the viral genome, amplification of the first library, fragmentation of the library, amplification of the second library using non-specific primer sets, and direct sequencing of the amplicon. To confirm the viruses detected by this method, we performed conventional PCR using virus-specific primers. We detected pathogenic virus genome sequences from the swab samples and confirmed infection with these viruses. In addition, we directly detected a viral genome sequence from the nasal swab sample without the viral propagation step. The RDV method is infrequently used in forensic analysis. This method is practicable with equipment existing in a normal laboratory and is useful for rapid detection and identification of pathogenic viruses in forensic samples. This method would also be applicable to the detection of bacteria and fungi.


Assuntos
Genoma Viral/genética , Cavidade Nasal/virologia , Faringe/virologia , Animais , Caliciviridae/genética , Gatos , DNA Viral , Herpesviridae/genética , Reação em Cadeia da Polimerase , RNA Viral , Manejo de Espécimes
2.
Forensic Sci Int ; 257: 376-378, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26528669

RESUMO

This study describes the validation of sampling techniques used to detect biological warfare agents used in terror attacks. For this purpose, we tested the efficiencies of different sampling media and extraction solutions for the recovery of bacterial pathogens. We first used Bacillus cereus ATCC 4342 spores as a surrogate for highly pathogenic B. anthracis to compare recovery efficiencies of spores from four different surfaces. We used three different types of sampling swabs and four different solutions to extract spores from the swabs. The most effective sampling method employed rayon swabs moistened with water. The efficencies of the four extraction solutions did not differ significantly, although yields were highest using phosphate-buffered saline containing Tween 80 (PBS-T). Using rayon swabs and sterile water, we recovered B. cereus ATCC 4342 and B. anthracis spores with equivalent efficiencies. These findings indicate that because of its reduced pathogenicity and relative ease in handling (Biosafety Level 1), use of B. cereus ATCC 4342 will facilitate further optimization of techniques to detect B. anthracis.


Assuntos
Bacillus anthracis/isolamento & purificação , Manejo de Espécimes/instrumentação , Manejo de Espécimes/métodos , Esporos Bacterianos/isolamento & purificação , Bacillus cereus/isolamento & purificação , Bioterrorismo , Tampões (Química) , Celulose , Humanos , Fosfatos , Poliésteres , Cloreto de Sódio , Água
3.
J Forensic Leg Med ; 34: 29-33, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-26165655

RESUMO

Forensic microbial specimens, including bacteria and viruses, are collected at biocrime and bioterrorism scenes. Although it is preferable that the pathogens in these samples are alive and kept in a steady state, the samples may be stored for prolonged periods before analysis. Therefore, it is important to understand the effects of storage conditions on the pathogens contained within such samples. To evaluate the capacity to preserve viable virus and the viral genome, influenza virus was added to the transport medium of the Universal Viral Transport system and stored for over 3 months at various temperatures, after which virus titrations and quantitative analysis of the influenza hemagglutinin gene were performed. Although viable viruses became undetectable 29 days after the medium was stored at room temperature, viruses in the medium stored at 4°C were viable even after 99 days. A quantitative PCR analysis indicated that the hemagglutinin gene was maintained for 99 days at both 4°C and room temperature. Therefore, long-term storage at 4°C has little effect on viable virus and viral genes, so the Universal Viral Transport system can be useful for microbial forensics. This study provides important information for the handling of forensic virus specimens.


Assuntos
Orthomyxoviridae/isolamento & purificação , Manejo de Espécimes/métodos , Hemaglutininas Virais/genética , Humanos , Viabilidade Microbiana , Reação em Cadeia da Polimerase , RNA Viral , Reação em Cadeia da Polimerase em Tempo Real , Temperatura
4.
J Forensic Leg Med ; 20(8): 1035-40, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24237815

RESUMO

Microbial forensic specimens that are collected at biocrime and bioterrorism scenes include blood, tissue, cloths containing biological fluids, swabs, water, soil, and aerosols. It is preferable that pathogens in such specimens are alive and kept in a steady state. Specimens may be stored for a prolonged period before analysis; therefore, it is important to understand the effect of the storage conditions on the pathogens contained within the specimens. In this study, we prepared blood and bloodstain specimens containing Gram-negative or -positive bacteria, stored the samples for 482 days under various conditions, and measured viable bacterial counts and total bacterial contents in the samples. Viable bacteria were preserved well in the samples stored at -30 and -80 °C, but were diminished or undetectable in the samples stored at 4 °C and room temperature. The total bacterial content was maintained in the blood samples stored at -30 and -80 °C and in the bloodstain samples stored under all temperature conditions, but decreased in the blood samples stored at 4 °C and room temperature. This study showed that the storage conditions affected viable bacteria and bacterial DNA and that freezing and drying were significant for their long-term storage. We provide important information for the storage of microbial forensic specimens.


Assuntos
Manchas de Sangue , DNA Bacteriano/sangue , Manejo de Espécimes , Temperatura , Carga Bacteriana , DNA Bacteriano/isolamento & purificação , Escherichia coli/genética , Escherichia coli/fisiologia , Medicina Legal , Humanos , Reação em Cadeia da Polimerase em Tempo Real , Streptococcus pyogenes/genética , Streptococcus pyogenes/fisiologia
5.
Leg Med (Tokyo) ; 13(6): 309-13, 2011 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-21940190

RESUMO

Nasal secretion has been regarded as one of the most difficult body fluids to identify and is especially difficult to discriminate from vaginal secretions and saliva. At present, few specific markers are known for nasal secretions. The aim of this study is to find a new approach for the identification of nasal secretions. We examined expression levels of statherin and histatin, peptides which are commonly found in saliva, in nasal and vaginal secretions by real-time RT-PCR and ELISA assays. Statherin mRNA was highly expressed in all nasal samples (dCt value=-1.49±1.10, n=8) and was detected even in 1-day-old 0.1-µL stains. However, the stability of mRNA in nasal stains was significantly (P<0.01) lower than in saliva. Low levels of statherin mRNA were detected in 4 of the 17 vaginal samples (dCt value=11.65-14.72). Histatin mRNA was not detected in any nasal or vaginal samples, although it was highly expressed in all saliva samples. ELISA assays with anti-statherin goat polyclonal antibody showed that statherin peptide was detected in all nasal and saliva samples even after dilution of more than 1000-fold. The statherin peptide was not detected in any vaginal samples, including samples that expressed low levels of statherin mRNA. The amount of statherin peptide in vaginal samples might be less than the limit of detection of this assay. In the present study, statherin was highly expressed in nasal secretions, but histatin was not. These markers may be useful for discriminating nasal secretions from vaginal secretions and saliva. However, the usefulness of these markers in practical forensic case samples has not yet been examined. Therefore, further research is required to establish the utility of these assays for identification of nasal secretions.


Assuntos
Expressão Gênica , Histatinas/análise , Mucosa Nasal/metabolismo , RNA Mensageiro/análise , Proteínas e Peptídeos Salivares/análise , Vagina/metabolismo , Biomarcadores/análise , Líquidos Corporais/química , Ensaio de Imunoadsorção Enzimática , Feminino , Medicina Legal/métodos , Histatinas/genética , Humanos , Proteínas/genética , Reação em Cadeia da Polimerase em Tempo Real , Proteínas e Peptídeos Salivares/genética
6.
Microbiol Res ; 166(2): 77-86, 2011 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-20347283

RESUMO

Molecular typing is an important tool in the surveillance and investigation of human Legionella infection outbreaks. In this study, two molecular typing methods, pulsed-field gel electrophoresis (PFGE) and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS), were used to discriminate 23 Legionella pneumophila strains. The usefulness of MALDI-TOF-MS was demonstrated. The MALDI-TOF-MS fingerprinting with filtered small acid-soluble molecules gave different molecular profiles among strains, and the clustal analysis with MALDI-TOF-MS showed a high discrimination of strains the same as that with PFGE. In addition, MALDI-TOF-MS data could be generated within a few hours after the initial culture, although PFGE analyses took several days to complete. Thus, MALDI-TOF-MS offers a simple and rapid discrimination technique that could aid in the tracking of fast-spreading outbreaks of Legionella.


Assuntos
Legionella pneumophila/classificação , Tipagem Molecular/métodos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Análise por Conglomerados , Eletroforese em Gel de Campo Pulsado/métodos , Humanos , Legionella pneumophila/genética , Legionella pneumophila/isolamento & purificação , Reação em Cadeia da Polimerase
7.
Int J Legal Med ; 124(5): 493-8, 2010 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-19953261

RESUMO

Statherin is a low molecular-weight phosphoprotein secreted from the parotid gland. Statherin mRNA was previously reported to be a useful marker for mRNA-based saliva identification. In this study, applicability of ELISA detection of statherin for forensic identification of saliva was investigated. The specificity and sensitivity of ELISA for detection of statherin were compared with those of ELISA for α-amylase and the Phadebas® amylase test. Statherin was specifically detected in saliva but not in other body fluids. In addition, statherin was successfully detected in aged saliva stains, mixed body fluids-saliva stains, and simulated casework samples. On the other hand, although ELISA for α-amylase showed higher sensitivity than ELISA for statherin, it was not specific enough to identify saliva. The Phadebas® amylase test also showed positive results in other body fluids that are known to have α-amylase activity; however, it is easy to use for screening forensic casework samples. In conclusion, ELISA for detection of statherin developed in this study could be an effective tool for the forensic identification of saliva because of its specificity for saliva among other body fluids. Forensic casework samples should be tested by ELISA detection or mRNA-based analysis for statherin, depending on the condition of the sample, to supplement presumptive tests for α-amylase, such as the Phadebas® amylase test.


Assuntos
RNA Mensageiro/metabolismo , Saliva/metabolismo , Proteínas e Peptídeos Salivares/genética , Biomarcadores/metabolismo , Análise Química do Sangue , Ensaio de Imunoadsorção Enzimática , Feminino , Medicina Legal , Humanos , Masculino , Sêmen/metabolismo , Sensibilidade e Especificidade , Suor/metabolismo , Urina/química , Vagina/metabolismo , alfa-Amilases/genética
8.
Biochem Biophys Res Commun ; 363(3): 531-5, 2007 Nov 23.
Artigo em Inglês | MEDLINE | ID: mdl-17888883

RESUMO

Bacillus anthracis causes anthrax, a lethal disease affecting humans, which has attracted attention due to its bioterrorism potential. gamma-Phage specifically infects B. anthracis, and is used for its detection. gamma-Phage lysin, PlyG, specifically lyses B. anthracis. Mutational analysis of PlyGB (PlyG binding domain; residues 156-233) indicated that positions 190-199 are necessary for binding to B. anthracis. This region is the central part of PlyGB and is predicted to form a beta-sheet. The amino acid residues of this region are also conserved in other lysins specific for B. anthracis. Alanine substitution at position 190 or 199 within this region resulted in significantly reduced binding, suggesting that L190 and Q199 play key roles in binding of PlyGB to B. anthracis. Our observations provide new insight into the mechanism of specific binding of lysin to B. anthracis, and may be useful in establishing new methods for detection of B. anthracis.


Assuntos
Aminoácidos/metabolismo , Fagos Bacilares/metabolismo , Bacillus anthracis/metabolismo , N-Acetil-Muramil-L-Alanina Amidase/metabolismo , Proteínas Virais/metabolismo , Sequência de Aminoácidos , Aminoácidos/genética , Fagos Bacilares/genética , Bacillus anthracis/virologia , Bacteriólise , Sítios de Ligação/genética , Cinética , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Proteínas Mutantes/metabolismo , Mutação , N-Acetil-Muramil-L-Alanina Amidase/genética , Ligação Proteica/genética , Homologia de Sequência de Aminoácidos , Proteínas Virais/genética
9.
Microbiol Immunol ; 51(2): 163-9, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-17310083

RESUMO

Detection of biological weapons is a primary concern in force protection, treaty verification, and safeguarding civilian populations against domestic terrorism. One great concern is the detection of Bacillus anthracis, the causative agent of anthrax. Therefore, there is a pressing need to develop novel methods for rapid, simple, and precise detection of B. anthracis. Here, we report that the C-terminal region of gamma-phage lysin protein (PlyG) binds specifically to the cell wall of B. anthracis and the recombinant protein corresponding to this region (positions, 156-233), PlyGB, is available as a bioprobe for detection of B. anthracis. Our detection method, based on a membrane direct blot assay using recombinant PlyGB, was more rapid and sensitive than the gamma-phage test and was simpler and more inexpensive than genetic methods such as PCR, or immunological methods using specific antibodies. Furthermore, its specificity was comparable to the gamma-phage test. PlyGB is applicable in conventional methods instead of antibodies and could be a potent tool for detection of B. anthracis.


Assuntos
Antraz/microbiologia , Bacillus anthracis/isolamento & purificação , Bacillus anthracis/metabolismo , N-Acetil-Muramil-L-Alanina Amidase/metabolismo , Proteínas Virais/metabolismo , Sequência de Aminoácidos , Antraz/diagnóstico , Antraz/prevenção & controle , Fagos Bacilares/genética , Cápsulas Bacterianas/fisiologia , Guerra Biológica , Parede Celular/metabolismo , DNA/química , DNA/genética , Immunoblotting , Microscopia Eletrônica de Transmissão , Dados de Sequência Molecular , N-Acetil-Muramil-L-Alanina Amidase/química , Reação em Cadeia da Polimerase , Ligação Proteica , Proteínas Recombinantes/metabolismo , Esporos Bacterianos/isolamento & purificação , Esporos Bacterianos/metabolismo , Proteínas Virais/química
10.
J Med Microbiol ; 54(Pt 3): 225-33, 2005 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-15713605

RESUMO

Interleukin 8 (IL8) is usually produced in both epithelial and monocytic cells during bacterial infections, causing inflammation. Helicobacter pylori induces production of IL8 from gastric epithelial cells via its cag pathogenicity island (cag PAI) system, LPS and outer-membrane protein. In some bacteria, heat-shock protein 60 (HSP60) also elicits a strong pro-inflammatory response in cells of the innate immune system. Three recombinant H. pylori HSP60 (rHSP60) proteins of different sizes were produced and one of these was used to raise two monoclonal antibodies (2E7 and 7B5). IL8 production was found to be induced in cultured monocytic cells treated with H. pylori cells or rHSP60 proteins, as measured by ELISA, and the amount of IL8 produced was dose-dependent. Pre-incubation of H. pylori cells or rHSP60 preparations with the antibody 2E7 significantly inhibited IL8 production from monocytic cells. These results indicated that HSP60 is closely associated with IL8 production in monocytic cells.


Assuntos
Chaperonina 60/metabolismo , Helicobacter pylori/metabolismo , Interleucina-8/biossíntese , Monócitos/metabolismo , Animais , Anticorpos Monoclonais/imunologia , Especificidade de Anticorpos , Linhagem Celular , Chaperonina 60/genética , Chaperonina 60/imunologia , Feminino , Helicobacter pylori/imunologia , Humanos , Interleucina-8/antagonistas & inibidores , Interleucina-8/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Monócitos/imunologia , Monócitos/microbiologia , Peptídeos/síntese química , Peptídeos/imunologia , RNA Mensageiro/biossíntese , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Células U937
11.
FEMS Immunol Med Microbiol ; 43(2): 125-31, 2005 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-15681141

RESUMO

Immune reactions against microorganisms play an important pathogenic role in Adamantiades-Behçet's disease. We had previously obtained Streptococcus sanguinis (strain BD113-20), isolated from the oral cavity of patients with Adamantiades-Behçet's syndrome. To investigate the pathogenesis of this isolate, we examined neutrophil reactions and levels of cytokine production by lymphocytes after stimulation with the strain. The reactions of neutrophils were examined by chemiluminescence assay using whole blood. The amounts of interferon gamma (IFN-gamma) and interleukin (IL)-4, IL-8, IL-10 and IL-12, produced by peripheral blood mononuclear cells, were measured by ELISA. Strain BD113-20 activated neutrophils from Adamantiades-Behçet's patients and healthy volunteers, and, in addition it increased the IFN-gamma production by lymphocytes. Lymphocytes from Adamantiades-Behçet's patients showed a dominant T helper-1 immune response. Results indicated that both bacterial stimulation and host hypersensitivity might be involved in the symptoms and pathogenesis of Adamantiades-Behçet's disease.


Assuntos
Síndrome de Behçet/imunologia , Linfócitos/imunologia , Neutrófilos/imunologia , Infecções Estreptocócicas/imunologia , Streptococcus sanguis/imunologia , Adulto , Idoso , Células Cultivadas , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Hipersensibilidade , Interferon gama/análise , Interleucinas/análise , Medições Luminescentes , Masculino , Pessoa de Meia-Idade , Boca
12.
Microbiology ; 150(Pt 12): 3913-22, 2004 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-15583145

RESUMO

Contact between Helicobacter pylori and gastric epithelial cells results in activation of NF-kappaB followed by secretion of interleukin (IL)-8. However, host-cell receptor(s) and their ligands involved in H. pylori-related IL-8 production have yet to be fully defined. In this study, the interaction between Toll-like receptors (TLRs), which are host receptors for pathogens involved in the innate immune response, and heat-shock protein (HSP) 60, an immune-potent antigen of H. pylori, was examined during H. pylori-induced IL-8 secretion in vitro. Recombinant H. pylori HSP60 (rHpHSP60) was prepared and added to cultured KATO III human gastric epithelial cells with or without pre-incubation with mouse monoclonal anti-TLR2 or anti-TLR4 antibodies. IL-8 mRNA expression and IL-8 protein release were analysed by Northern blotting and immunoassay. Involvement of NF-kappaB activation was analysed immunocytochemically by anti-NF-kappaB p65 antibody and ammonium pyrrolidinedithiocarbamate (PDTC), an inhibitor of NF-kappaB-mediated transcriptional activation. rHpHSP60 induced IL-8 mRNA expression and IL-8 secretion in a dose-dependent manner in KATO III cells. Anti-TLR2 antibody inhibited rHpHSP60-induced IL-8 secretion by 75 %, and anti-TLR4 antibody inhibited it by 30 %. rHpHSP60 induced nuclear translocation of NF-kappaB p65, which was inhibited by pretreatment with anti-TLR2 antibody. Treatment with PDTC significantly decreased the secretion of IL-8 induced by rHpHSP60. These findings suggest that H. pylori HSP60 activates NF-kappaB and induces IL-8 production through TLR-triggered pathways in gastric epithelial cells. Thus, it is possible that H. pylori HSP60 and TLR interaction in host cells contributes to the development of gastric inflammation caused by H. pylori infection.


Assuntos
Chaperonina 60/metabolismo , Células Epiteliais/microbiologia , Mucosa Gástrica/microbiologia , Helicobacter pylori/patogenicidade , Interleucina-8/metabolismo , Glicoproteínas de Membrana/metabolismo , Receptores de Superfície Celular/metabolismo , Animais , Linhagem Celular , Chaperonina 60/genética , Mucosa Gástrica/citologia , Helicobacter pylori/imunologia , Humanos , Imuno-Histoquímica , Inflamação/etiologia , Camundongos , NF-kappa B/metabolismo , Receptor 2 Toll-Like , Receptor 4 Toll-Like , Receptores Toll-Like
13.
J Med Microbiol ; 53(Pt 3): 207-12, 2004 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-14970245

RESUMO

To date, two Helicobacter species, Helicobacter pylori and 'Helicobacter heilmannii' (formerly named 'Gastrospirillum hominis'), have been identified from the human stomach. In this study, we observed non-H. pylori-shaped bacteria in gastric tissue sections and successfully isolated them by cultivation. Elongated bacteria were isolated from a patient with gastric-mucosa-associated lymphoid-tissue lymphoma who had been diagnosed as H. pylori-negative by culture, rapid urease test and histopathology in another hospital. The bacteria were grown only on chocolate agar in a CO2 incubator, appeared more than 10 microm long in histological sections, formed small colonies and showed poor growth in a brain heart infusion broth; these characteristics apparently differed from common clinical isolates of H. pylori. However, the bacteria were identified as H. pylori by PCR of the urease gene, 16S rDNA sequencing, protein profile and antigenicity examined by anti-H. pylori polyclonal antibody. These observations suggest that the H. pylori strain identified in this study may contribute to the development of gastroduodenal diseases in cases judged as H. pylori-negative by ordinary methods.


Assuntos
Mucosa Gástrica/microbiologia , Helicobacter pylori/isolamento & purificação , Linfoma de Zona Marginal Tipo Células B/microbiologia , Proteínas de Bactérias/análise , Proteínas de Bactérias/imunologia , Western Blotting , DNA Bacteriano/análise , DNA Ribossômico/análise , Mucosa Gástrica/patologia , Helicobacter pylori/classificação , Helicobacter pylori/genética , Helicobacter pylori/ultraestrutura , Humanos , Microscopia Eletrônica , Microscopia Eletrônica de Varredura , Reação em Cadeia da Polimerase , RNA Ribossômico 16S/genética , Urease/genética
14.
Biosci Biotechnol Biochem ; 66(8): 1628-34, 2002 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-12353620

RESUMO

The bioavailability of a series of novel acylated ascorbic acid derivatives, 6-O-acyl-2-O-alpha-D-glucopyranosyl-L-ascorbic acids (6-Acyl-AA-2G), as an ascorbic acid (AA) supplement was investigated in rats and guinea pigs. Oral administration of 6-Acyl-AA-2G to rats resulted in an increase in the plasma AA level. However, the intact form was not detectable in the plasma by high-performance liquid chromatography, indicating its hydrolysis through the process of absorption. After an intravenous injection to rats of 6-Octa-AA-2G as a representative derivative, the intact form rapidly disappeared from the plasma, being followed by a prolonged and marked elevation of the plasma AA level. Various tissue homogenates from guinea pigs were examined for their releasing activity of AA, 2-O-alpha-D-glucopyranosyl-L-ascorbic acid (AA-2G) and 6-O-acyl-AA from 6-Acyl-AA-2G. High activity was observed in the small intestine. These hydrolytic activities to AA and 6-O-acyl-AA were completely inhibited by castanospermine, an alpha-glucosidase inhibitor, and AA-2G was observed as the only resulting hydrolysate, suggesting the participation of alpha-glucosidase and esterase in the in vivo hydrolysis of 6-Acyl-AA-2G. 6-Octa-AA-2G was found to exhibit an obvious therapeutic effect in scorbutic guinea pigs from its repeated oral administration. These results indicate that 6-Acyl-AA-2G is a readily available source of AA activity in vivo, and may be useful as an effective pharmacological agent and as a promising food additive.


Assuntos
Ácido Ascórbico/análogos & derivados , Ácido Ascórbico/farmacocinética , Animais , Ácido Ascórbico/sangue , Deficiência de Ácido Ascórbico/tratamento farmacológico , Deficiência de Ácido Ascórbico/metabolismo , Disponibilidade Biológica , Encéfalo/metabolismo , Suplementos Nutricionais , Esterases/metabolismo , Cobaias , Hidrólise , Intestino Delgado/metabolismo , Fígado/metabolismo , Masculino , Ratos , Ratos Wistar , Escorbuto/metabolismo , Pele/metabolismo , alfa-Glucosidases/metabolismo
15.
J Med Chem ; 45(2): 462-8, 2002 Jan 17.
Artigo em Inglês | MEDLINE | ID: mdl-11784150

RESUMO

A series of novel monoacylated vitamin C derivatives were chemically synthesized with a stable ascorbate derivative, 2-O-alpha-D-glucopyranosyl-L-ascorbic acid (AA-2G), and acid anhydrides in pyridine. Their solubility in organic phase, thermal stability, radical scavenging activity, and in vitro skin permeability was evaluated. These monoacylated derivatives were identified as 6-O-acyl-2-O-alpha-D-glucopyranosyl-L-ascorbic acids (6-Acyl-AA-2G) by UV spectra, elemental analyses, and nuclear magnetic resonance spectroscopy. The reactions afforded 6-Acyl-AA-2G in high yields (30-60%). 6-Acyl-AA-2G exhibited satisfactory stability in neutral solution comparable to that of a typical stable derivative, AA-2G, and also showed the radical scavenging activity. The lipid solubility of 6-Acyl-AA-2G was increased with increasing length of their acyl group. Increased skin permeability was superior to those of AA-2G and ascorbic acid (AsA). 6-Acyl-AA-2G that is susceptible to enzymatic hydrolysis by tissue esterase and/or alpha-glucosidase produces AA-2G and AsA, which is in the skin tissues. Thus, these findings indicate that the novel vitamin C derivatives presented here, 6-Acyl-AA-2G, may be effective antioxidants in skin care and medicinal use.


Assuntos
Antioxidantes/síntese química , Ácido Ascórbico/análogos & derivados , Ácido Ascórbico/síntese química , Depuradores de Radicais Livres/síntese química , Glucosídeos/síntese química , Pele/metabolismo , 1-Octanol , Antioxidantes/química , Antioxidantes/farmacologia , Ácido Ascórbico/química , Ácido Ascórbico/farmacologia , Cromatografia Líquida de Alta Pressão , Esterases/metabolismo , Depuradores de Radicais Livres/química , Depuradores de Radicais Livres/farmacologia , Glucosídeos/química , Glucosídeos/farmacologia , Humanos , Hidrólise , Técnicas In Vitro , Permeabilidade , Solubilidade , Espectrofotometria Ultravioleta , Relação Estrutura-Atividade , Água , alfa-Glucosidases/metabolismo
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA