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1.
J Am Chem Soc ; 146(7): 4532-4541, 2024 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-38326951

RESUMO

Here, we present the synthesis and characterization of a novel 2D crystalline framework, named C2O, which mainly consists of carbon and oxygen in a 2:1 molar ratio and features crown ether holes in its skeletal structure. The covalent-frameworked 2D crown ether can be synthesized on a gram-scale and exhibits fine chemical stability in various environments, including acid, base, and different organic solvents. The C2O efficiently activates KI through the strong coordination of K+ with crown ether holes in a rigid framework, which enhances the nucleophilicity of I- and significantly improves its catalytic activity for CO2 fixation with epoxides. The presence of C2O with KI results in remarkable increases in CO2 conversion from 5.7% to 99.9% and from 2.9% to 74.2% for epichlorohydrin and allyl glycidyl ether, respectively. Moreover, C2O possesses both electrophilic and nucleophilic sites at the edge of its framework, allowing for the customization of physicochemical properties by a diverse range of chemical modifications. Specifically, incorporating allyl glycidyl ether (AGE) as an electrophile or ethoxyethylamine (EEA) as a nucleophile into C2O enables the synthesis of C2O-AGE or C2O-EEA, respectively. These modified frameworks exhibit improved conversions of 97.2% and 99.9% for CO2 fixation with allyl glycidyl ether, outperforming unmodified C2O showing a conversion of 74.2%. This newly developed scalable, durable, and customizable covalent framework holds tremendous potential for the design and preparation of outstanding materials with versatile functionalities, rendering them highly attractive for a wide range of applications.

2.
Vet Res ; 55(1): 87, 2024 Jul 09.
Artigo em Inglês | MEDLINE | ID: mdl-38982477

RESUMO

Hepatitis E virus (HEV) is a major cause of viral hepatitis worldwide. Pigs are the natural host of HEV genotype 3 and the main reservoir of HEV. As the host range of HEV genotype 3 expands, the possibility that HEV from various species can be transmitted to humans via pigs is increasing. We investigated the potential cross-species transmission of HEV by infecting minipigs with swine HEV (swHEV), rabbit HEV (rbHEV), and human HEV (huHEV) and examining their histopathological characteristics and distribution in various organs. Fifteen specific-pathogen-free Yucatan minipigs were infected with swHEV, rbHEV, huHEV, or a mock control. In the present study, we analysed faecal shedding, viremia, and serological parameters over a seven-week period. Our results indicated that swHEV exhibited more robust shedding and viremia than non-swHEVs. Only swHEV affected the serological parameters, suggesting strain-specific differences. Histopathological examination revealed distinct patterns in the liver, pancreas, intestine, and lymphoid tissues after infection with each HEV strain. Notably, all three HEVs induced histopathological changes in the pancreas, supporting the association of HEVs with acute pancreatitis. Our results also identified skeletal muscle as a site of HEV antigen presence, suggesting a potential link to myositis. In conclusion, this study provides valuable insights into the infection dynamics of different HEV strains in minipigs, emphasizing the strain-specific variations in virological, serological, and histological parameters. The observed differences in infection kinetics and tissue tropism will contribute to our understanding of HEV pathogenesis and the potential for cross-species transmission.


Assuntos
Vírus da Hepatite E , Hepatite E , Doenças dos Suínos , Porco Miniatura , Animais , Suínos , Hepatite E/veterinária , Hepatite E/virologia , Hepatite E/transmissão , Vírus da Hepatite E/fisiologia , Doenças dos Suínos/virologia , Doenças dos Suínos/transmissão , Doenças dos Suínos/patologia , Organismos Livres de Patógenos Específicos , Coelhos , Eliminação de Partículas Virais , Humanos , Fezes/virologia , Feminino , Viremia/veterinária , Viremia/virologia
3.
Foodborne Pathog Dis ; 16(6): 411-420, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30920321

RESUMO

Contamination of fresh vegetables and berries with human enteric viruses is a major cause of food poisoning. The aim of this study was to investigate the prevalence of norovirus GI, norovirus GII, hepatitis A virus (HAV), adenovirus, astrovirus, rotavirus, and male-specific coliphage systematically in fresh fruit and vegetables and associated agricultural environmental samples, including irrigation water, soil, and worker's gloves. Enteric viruses were detected by international standard methods (ISO/TS 15216), and male-specific coliphages were isolated using US EPA Method 1601. For the study, 773 samples were collected from June 2016 to April 2017, including Chinese cabbage (n = 244), cucumber (n = 98), lettuce (n = 73), strawberry (n = 120), soil (n = 191), irrigation water (n = 14), and gloves (n = 27). Two cucumber and two irrigation water samples were positive for norovirus GI, and one cucumber and two irrigation water samples were positive for norovirus GII. HAV was detected in one strawberry sample and one glove sample. The other tested foodborne viruses were not detected in any of the samples. Sixteen male-specific coliphages were isolated from Chinese cabbage, cucumber, lettuce, cherry tomato, soil, and irrigation water. The isolation of male-specific coliphage would be more practical to investigate the fecal contamination in produce rather than pathogenic viruses.


Assuntos
Microbiologia de Alimentos , Frutas/virologia , Verduras/virologia , Vírus/isolamento & purificação , Irrigação Agrícola , Demografia , Enterovirus/isolamento & purificação , Contaminação de Alimentos , Vírus da Hepatite A/isolamento & purificação , Humanos , Norovirus/isolamento & purificação , República da Coreia , Rotavirus/isolamento & purificação
4.
J Med Virol ; 90(4): 655-662, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29106738

RESUMO

Ten Yucatan miniature piglets were challenged with the human norovirus (NoV) GII.12/GII.3 CAU140599 strain and five piglets were used as negative controls. Stool, serum, and organs were collected and processed from two NoV-infected piglets and one negative piglet at 1, 2, 3, 5, and 7 days post-inoculation (dpi). NoV was detected in stool and serum samples by real-time RT-PCR. Mild diarrhea was observed at 1-3 dpi. Fecal shedding and viremia were detected intermittently at 1, 3, and 7 dpi. While interferon-α was significantly elevated at 2-3 dpi, interferon-γ was not changed. Immunohistochemistry demonstrated that the NoV capsid antigen was present in macrophages, lymphocytes, and dendritic cells of the stomach, intestines, lymph nodes, spleen, and tonsils. Intestinal epithelium did not exhibit a positive signal for NoV. In addition, negative-sense viral RNA was confirmed in immune cells by fluorescence in situ hybridization. Therefore, NoV might be associated with macrophages and lymphocytes in gastrointestinal tract and immune organs of experimentally infected miniature piglets.


Assuntos
Infecções por Caliciviridae/patologia , Modelos Animais de Doenças , Genótipo , Norovirus/patogenicidade , Estruturas Animais/patologia , Estruturas Animais/virologia , Animais , Animais Recém-Nascidos , Diarreia/patologia , Fezes/virologia , Imuno-Histoquímica , Linfócitos/virologia , Macrófagos/virologia , Norovirus/classificação , Norovirus/genética , Norovirus/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real , Suínos , Porco Miniatura , Fatores de Tempo , Eliminação de Partículas Virais
5.
Korean J Parasitol ; 56(4): 371-374, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30196670

RESUMO

A 3-month-old female Maltese puppy was hospitalized with persistent diarrhea in a local veterinary clinic. Blood chemistry and hematology profile were analyzed and fecal smear was examined. Diarrheal stools were examined in a diagnostic laboratory, using multiplex real-time polymerase chain reaction (PCR) against 23 diarrheal pathogens. Sequence analysis was performed using nested PCR amplicon of 18S ribosomal RNA. Coccidian oocysts were identified in the fecal smear. Although multiplex real-time PCR was positive for Cyclospora cayetanensis, the final diagnosis was Cystoisospora ohioensis infection, confirmed by phylogenetic analysis of 18S rRNA. To our knowledge, this the first case report of C. ohioensis in Korea, using microscopic examination and phylogenetic analysis.


Assuntos
Coccídios/isolamento & purificação , Coccidiose/parasitologia , Coccidiose/veterinária , Diarreia/parasitologia , Diarreia/veterinária , Doenças do Cão/parasitologia , Enteropatias Parasitárias/parasitologia , Enteropatias Parasitárias/veterinária , Oocistos/isolamento & purificação , Animais , Coccídios/genética , Cães , Fezes/parasitologia , Feminino , Filogenia , Reação em Cadeia da Polimerase/métodos , RNA Ribossômico 18S , República da Coreia
6.
J Clin Microbiol ; 52(2): 557-63, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24478488

RESUMO

This study aimed to develop a loop-mediated isothermal amplification (LAMP) method for the rapid detection of Arcobacter species. Specific primers targeting the 23S ribosomal RNA gene were used to detect Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii. The specificity of the LAMP primer set was assessed using DNA samples from a panel of Arcobacter and Campylobacter species, and the sensitivity was determined using serial dilutions of Arcobacter species cultures. LAMP showed a 10- to 1,000-fold-higher sensitivity than multiplex PCR, with a detection limit of 2 to 20 CFU per reaction in vitro. Whereas multiplex PCR showed cross-reactivity with Campylobacter species, the LAMP method developed in this study was more sensitive and reliable than conventional PCR or multiplex PCR for the detection of Arcobacter species.


Assuntos
Arcobacter/isolamento & purificação , Microbiologia de Alimentos , Infecções por Bactérias Gram-Negativas/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Animais , Arcobacter/genética , Primers do DNA/genética , DNA Bacteriano/genética , DNA Ribossômico/genética , Infecções por Bactérias Gram-Negativas/microbiologia , Humanos , RNA Ribossômico 23S/genética , Sensibilidade e Especificidade
7.
Food Microbiol ; 44: 81-6, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25084649

RESUMO

Fermented seafood is popular in Asian countries. This study examined the survival of feline calicivirus (FCV) and murine norovirus (MNV) during oyster fermentation. Oysters spiked with FCV and MNV were fermented with 5% or 10% salt at 18 °C for 15 days, and MNV and FCV titers, lactic acid bacteria (LAB) populations, pH, and enzymatic activity were measured at 0, 1, 3, 5, 7, 10, and 15 days post-fermentation (DPF). Reductions in MNV and FCV were greater in 5% NaCl-supplemented oysters than in 10% NaCl-supplemented oysters. In 5% NaCl oysters, MNV and FCV titers significantly decreased by 1.60 log and 3.01 log, respectively, at 15 DPF. Populations of LAB increased from 3.62 log10 colony-forming units/g at 0 DPF to 8.77 log10 colony-forming units/g at 15 DPF during oyster fermentation supplemented with 5% NaCl supplementation, and the pH decreased gradually from 5.38 at 0 DPF to 4.17 at 15 DPF. During oyster fermentation, α-amylase, proteinase, and lipase were produced at higher levels in 5% salted oysters than in 10% salted oysters (P < 0.01). We concluded that many of the antimicrobial factors produced in fermented oysters could contribute to a reduction in foodborne viruses.


Assuntos
Calicivirus Felino/fisiologia , Norovirus/fisiologia , Ostreidae/microbiologia , Alimentos Marinhos/microbiologia , Inativação de Vírus , Animais , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Bactérias/metabolismo , Gatos , Linhagem Celular , Fermentação , Concentração de Íons de Hidrogênio , Ácido Láctico/metabolismo , Camundongos , Ostreidae/química , Alimentos Marinhos/análise
8.
Front Vet Sci ; 9: 913622, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36246307

RESUMO

Emerging infectious diseases (EID) in humans and animals are proving to be a serious health concern. This study investigated the prevalence of emerging or re-emerging human enteric viruses in porcine stools and swabs. Eleven enteric EID viruses were selected as target viruses for the current study and ranked based on their impact on public health and food safety: enterovirus (EV), hepatitis E virus, norovirus GI and GII, sapovirus (SaV), adenovirus (AdV), astrovirus, rotavirus, hepatitis A virus, aichivirus, and bocavirus. Using real-time RT-PCR or real-time PCR, EID viruses were detected in 129 (86.0%) of 150 samples. The most prevalent virus was EV, which was detected in 68.0% of samples, followed by AdV with a detection rate of 38.0%. In following sequencing and phylogenetic analyses, 33.0% (58/176) of the detected viruses were associated with human enteric EID viruses, including AdV-41, coxsackievirus-A2, echovirus-24, and SaV. Our results show that porcine stools frequently contain human enteric viruses, and that few porcine enteric viruses are genetically related to human enteric viruses. These findings suggest that enteric re-emerging or EID viruses could be zoonoses, and that continuous monitoring and further studies are needed to ensure an integrated "One Health" approach that aims to balance and optimize the health of humans, animals, and ecosystems.

9.
JAMA Surg ; 157(9): 828-834, 2022 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-35895073

RESUMO

Importance: Appendectomy remains the standard of care for uncomplicated acute appendicitis despite several randomized clinical trials pointing to the safety and efficacy of nonoperative management of this disease. A meta-analysis of randomized clinical trials may contribute to the body of evidence and help surgeons select which patients may benefit from surgical and nonsurgical treatment. Objective: To assess the efficacy and safety of nonoperative management vs appendectomy for acute uncomplicated appendicitis. Data Sources: A systematic review was conducted using indexed sources (Embase and PubMed) to search for published randomized clinical trials in English comparing nonoperative management with appendectomy in adult patients presenting with uncomplicated acute appendicitis. To increase sensitivity, no limits were set for outcomes reported, sex, or year of publication. All nonrandomized or quasi-randomized trials were excluded, and validated primers were used. Study Selection: Among 1504 studies imported for screening, 805 were duplicates, and 595 were excluded for irrelevancy. A further 96 were excluded after full-text review, mainly owing to wrong study design or inclusion of pediatric populations. Eight studies met the inclusion criteria and were selected for the meta-analysis. Data Extraction and Synthesis: Meta-extraction was conducted with independent extraction by multiple reviewers using the Covidence platform for systematic reviews and in accordance with PRISMA guidelines. Data were pooled by a random-effects model. Main Outcomes and Measures: Treatment success and major adverse effects at 30 days' follow-up. Results: The main outcome (treatment success proportion at 30 days of follow-up) was not significantly different in the operative and nonoperative management cohorts (risk ratio [RR], 0.85; 95% CI, 0.66-1.11). Likewise, the percentage of major adverse effects was similar in both cohorts (RR, 0.72; 95% CI, 0.29-1.79). However, in the nonoperative management group, length of stay was significantly longer (RR, 1.48; 95% CI, 1.26-1.70), and a median cumulative incidence of 18% of recurrent appendicitis was observed. Conclusions and Relevance: These results point to the general safety and efficacy of nonoperative management of uncomplicated acute appendicitis. However, this strategy may be associated with an increase in duration of hospital stay and a higher rate of recurrent appendicitis. This meta-analysis may help inform decision-making in nonoperative management of uncomplicated acute appendicitis.


Assuntos
Apendicite , Doença Aguda , Adulto , Antibacterianos/uso terapêutico , Apendicectomia/efeitos adversos , Apendicite/complicações , Apendicite/cirurgia , Criança , Humanos , Resultado do Tratamento
10.
Viruses ; 13(2)2021 02 23.
Artigo em Inglês | MEDLINE | ID: mdl-33672228

RESUMO

Mushrooms are used in their natural form as a food supplement and food additive. In addition, several bioactive compounds beneficial for human health have been derived from mushrooms. Among them, polysaccharides, carbohydrate-binding protein, peptides, proteins, enzymes, polyphenols, triterpenes, triterpenoids, and several other compounds exert antiviral activity against DNA and RNA viruses. Their antiviral targets were mostly virus entry, viral genome replication, viral proteins, and cellular proteins and influenced immune modulation, which was evaluated through pre-, simultaneous-, co-, and post-treatment in vitro and in vivo studies. In particular, they treated and relieved the viral diseases caused by herpes simplex virus, influenza virus, and human immunodeficiency virus (HIV). Some mushroom compounds that act against HIV, influenza A virus, and hepatitis C virus showed antiviral effects comparable to those of antiviral drugs. Therefore, bioactive compounds from mushrooms could be candidates for treating viral infections.


Assuntos
Agaricales/química , Antivirais/farmacologia , Extratos Vegetais/farmacologia , Animais , Antivirais/química , Humanos , Extratos Vegetais/química , Viroses/tratamento farmacológico , Viroses/virologia , Replicação Viral/efeitos dos fármacos , Vírus/efeitos dos fármacos
11.
Microorganisms ; 8(8)2020 Aug 11.
Artigo em Inglês | MEDLINE | ID: mdl-32796772

RESUMO

This study aimed to investigate the prevalence of foodborne viruses in reservoirs (an important resource of irrigation water) and its correlation with environmental and weather factors. From May 2017 to November 2018, we visited ten reservoirs and a river in the Anseong region of South Korea and collected a total of 192 samples in accordance with the environment protection agency guidelines. We recorded the weather factors (temperature, humidity, and accumulated precipitation) and investigated the surrounding environment factors (livestock, fishing site, the catchment area of reservoirs, etc.). Our research results show that from the river and reservoirs, the detection rates of human norovirus GII, adenovirus, rotavirus, human norovirus GI, and astrovirus were 27.1, 10.4, 10.4, 4.16, and 3.1%, respectively. Their viral load ranged from -1.48 to 1.55 log10 genome copies/l. However, hepatitis A virus was not detected in any irrigation water sample. Although no sampling was performed in winter, foodborne viruses and male-specific coliphages were frequently found during spring (40.78%) and autumn (39.47%). Interestingly, the significant correlation between the accumulative precipitation and the number of detected norovirus and adenovirus was confirmed by linear regression analysis. Furthermore, when the accumulative precipitation ranged from 20 to 60 mm, it significantly affected the viral load and prevalence. Among the environmental factors, recreational facilities such as fishing sites and bungalow fishing spots were identified as contamination sources by correlation analysis. Our research results confirmed the correlations between environmental contamination factors in the reservoir and weather factors with the prevalence of foodborne viruses in the reservoir. These facilitates the assessment of potential foodborne virus contamination during crop irrigation. In addition, predictive models including environmental and weather factors should be developed for monitoring and controlling the safety of irrigation waters in reservoirs.

12.
Sci Rep ; 10(1): 12022, 2020 07 21.
Artigo em Inglês | MEDLINE | ID: mdl-32694702

RESUMO

Infection by hepatitis E virus (HEV) via the oral route causes acute hepatitis. Extra-hepatic manifestations of HEV infection may stem from various causes; however, its distribution in organs such as the liver, as well as the mechanisms underlying HEV-induced cell injury, remain unclear. The objective of this study was to determine the chronological distribution of HEV in various tissues of HEV-challenged miniature pigs and to investigate the mechanisms underlying HEV-induced cell death in the pancreas and liver. Virological and serological analyses were performed on blood and faecal samples. Histopathology of the liver and extra-hepatic tissues was analysed. Cell death pathways and immune cell characterisation in inflammatory lesions were analysed using immunohistochemistry. The liver and pancreas displayed inflammation and cellular injury, and a large amount of HEV was observed in the lesions. The liver was infiltrated by T and natural killer cells. HEV was identified in all organs except the heart, and was associated with immune cells. Although the liver and the pancreas strongly expressed TNF-α and TRAIL, TUNEL assay results were negative. RIP3 and pMLKL were expressed in the pancreas. RIP3, but not pMLKL, was expressed in the liver. Pancreatitis induced in HEV-infected miniature pigs is associated with necroptosis.


Assuntos
Vírus da Hepatite E/imunologia , Hepatite E/imunologia , Necroptose , Pâncreas/patologia , Doenças dos Suínos/imunologia , Animais , Modelos Animais de Doenças , Fezes/virologia , Hepatite E/complicações , Hepatite E/virologia , Vírus da Hepatite E/genética , Células Matadoras Naturais/imunologia , Fígado/imunologia , Fígado/patologia , Pâncreas/imunologia , Pancreatite/etiologia , Pancreatite/imunologia , Pancreatite/virologia , RNA Viral/análise , RNA Viral/sangue , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Suínos , Doenças dos Suínos/virologia , Porco Miniatura , Linfócitos T/imunologia
13.
Int J Food Microbiol ; 299: 58-63, 2019 Jun 16.
Artigo em Inglês | MEDLINE | ID: mdl-30954876

RESUMO

Bivalve molluscan shellfish, such as oysters, clams, and cockles, are well-recognized as vectors that concentrate foodborne pathogens by filter feeding. The objective of this study was to investigate the distribution and persistence of hepatitis A virus (HAV) in experimentally contaminated oysters that were either fed or not fed with algae. Oysters were experimentally contaminated with HAV and maintained in depuration conditions. qRT-PCR, immunohistochemistry (IHC), and in situ hybridization (ISH) were performed on oyster samples collected at 0, 1, 3, 5, and 7 days post-inoculation. When HAV-contaminated oysters were depurated for 7 days, HAV was detected in 91.1-97.8% of the digestive glands and gills. While the high viral load in the digestive glands in oysters did not change significantly regardless of algae-feeding, the viral load of the gills gradually decreased in both groups during the depuration. HAV antigen and RNA were detected in the digestive diverticula and connective tissues by both IHC and ISH. HAV was detected in the stomach, intestine, and gills by only ISH. The distribution of HAV in various oyster tissues may explain the persistence of contamination in oysters during the depuration process.


Assuntos
Microbiologia de Alimentos , Vírus da Hepatite A/fisiologia , Ostreidae/virologia , Animais , Manipulação de Alimentos , Trato Gastrointestinal/virologia , Brânquias/virologia , Vírus da Hepatite A/genética , Frutos do Mar/virologia , Fatores de Tempo
14.
Mitochondrial DNA A DNA Mapp Seq Anal ; 29(5): 674-678, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-28696810

RESUMO

Cases with three or more consecutive spontaneous abortions before the 20th week of gestation are termed as recurrent pregnancy loss (RPL). Problems in implantation of the foetus and any retarded growth of the foetus in the uterus can be correlated to RPL. Possible causes of RPL would include the genetic variations in the regulatory enzymes of the crucial metabolic pathways, clotting factors, hormones and hormone receptors. This defect of the mitochondrial respiratory chain is recognized as a major cause of human disease. We investigated 73 women with RPL and 100 healthy normal controls. By using the direct sequencing method, the amplified products including the mtDNA complex I genes were analyzed. Overall, seven variations in mitochondrial complex I genes were found (T4216C, A5153G, C10142T, C12062T, A12662G, G14179A and T14263C) using direct sequencing technique. The RPL group had significantly higher proportions of the different variants than those observed of the control group. In conclusion, more research is essentially needed to understand the effect and role of the mitochondrial variations in the progress of RPL, which may vary among individuals and different ethnic groups.


Assuntos
Aborto Habitual/genética , DNA Mitocondrial/genética , NADH Desidrogenase/genética , Adulto , Estudos de Casos e Controles , Complexo I de Transporte de Elétrons/genética , Feminino , Predisposição Genética para Doença , Genoma Mitocondrial , Humanos , Mitocôndrias/genética , Proteínas Mitocondriais/genética , Gravidez
15.
Anticancer Res ; 38(1): 71-76, 2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-29277758

RESUMO

BACKGROUND/AIM: Rhabdomyosarcoma is the most common type of pediatric soft-tissue sarcoma. Among the subsets of this disease, alveolar rhabdomyosarcoma (ARMS) expressing paired box 3 (PAX3) and forkhead box O1 (PAX3-FOXO1) fusion oncoprotein has the worst prognosis. The goal of this study was to investigate the chemotherapeutic effects of sphingosine on PAX3-FOXO1-positive ARMS cells [tumor protein p53 (TP53)-mutated RH30 and TP53 wild-type RH18 cells]. MATERIALS AND METHODS: The proliferation, cell death, apoptosis, cell cycle, and MYCN proto-oncogene (MYCN) expression of RH30 and RH18 cells were determined. RESULTS: Sphingosine inhibited the growth and caused cell death in a dose-dependent manner in both cell lines. Sphingosine triggered cell death by inducing apoptosis without affecting the cell cycle. MYCN expression was down-regulated within 2 and 4 h of sphingosine treatment in both RH30 and RH18 cells. CONCLUSION: Sphingosine exerts antiproliferative and pro-apoptotic effects via MYCN down-regulation independently of TP53 mutation status in PAX3-FOXO1-positive ARMS cells.


Assuntos
Antineoplásicos/farmacologia , Proteína Proto-Oncogênica N-Myc/genética , Rabdomiossarcoma Alveolar/genética , Esfingosina/farmacologia , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Regulação para Baixo/efeitos dos fármacos , Proteína Forkhead Box O1/metabolismo , Humanos , Mutação , Fator de Transcrição PAX3/metabolismo , Proto-Oncogene Mas , Rabdomiossarcoma Alveolar/metabolismo , Proteína Supressora de Tumor p53/genética
16.
Phytomedicine ; 30: 59-66, 2017 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-28545670

RESUMO

BACKGROUND: Human noroviruses (HuNoV), which are responsible for acute gastroenteritis, are becoming a serious public health concern worldwide. Since no effective antiviral drug or vaccine for HuNoV has been developed yet, some natural extracts and their active components have been investigated for their ability to inhibit noroviruses. However, their exact antiviral mechanisms have not been investigated. PURPOSE: This study was performed to investigate the expression of interferon (IFN)-α, IFN-λ, tumor necrosis factor-α (TNF-α), Mx, and zinc finger CCCH type antiviral protein 1 (ZAP), 2'-5' oligo (A) synthetase (OAS), and inducible nitric oxide synthase (iNOS) in RAW 264.7 cells pre-treated with fisetin, daidzein, quercetin, epigallocatechin gallate (EGCG), and epicatechin gallate (ECG) that have anti-noroviral activity. STUDY DESIGN: Based on the antiviral activity of the five flavonoids, recently reported by our group, the expression of antiviral factors such as IFN-α, IFN-λ, TNF-α, IL-1ß, IL-6, Mx, ZAP, OAS, and iNOS was investigated in RAW 264.7 cells pre-treated with these flavonoids. METHODS: Anti-noroviral effect was determined by performing a plaque assay on cells treated with the flavonoid. RAW 264.7 cells were treated with fisetin, daidzein, quercetin, EGCG, and ECG. Then, mRNA of IFN-α, IFN-λ, TNF-α, IL-1ß, IL-6, Mx, ZAP, OAS, and iNOS were measured by real-time RT-PCR. IFN-α, TNF-α, IL-1ß, and IL-6 proteins were measured by ELISA. RESULTS: Pre-treatment with fisetin (50µM), fisetin (100µM), EGCG (100µM), quercetin (100µM), daidzein (50µM), and ECG (150µM) significantly reduced MNoV by 50.00±7.14 to 60.67±9.26%. The mRNA levels of IFN-α, IFN-λ, TNF-α, Mx, and ZAP were upregulated in RAW 264.7 cells pre-treated with fisetin, quercetin, and daidzein, but not in those pre-treated with EGCG or ECG. Regarding protein levels, IFN-α was significantly induced in cells pre-treated with fisetin, quercetin, and daidzein, whereas TNF-α was significantly induced only in cells pre-treated with daidzein. CONCLUSION: Pre-treatment of RAW 264.7 cells with the five flavonoids inhibited MNoV by upregulating the expression of antiviral cytokines (IFN-α, IFN-λ, and TNF-α) and interferon-stimulating genes (Mx and ZAP).


Assuntos
Antivirais/farmacologia , Citocinas/metabolismo , Flavonoides/farmacologia , Norovirus/efeitos dos fármacos , Animais , Catequina/análogos & derivados , Catequina/farmacologia , Linhagem Celular , Citocinas/genética , Avaliação Pré-Clínica de Medicamentos/métodos , Flavonóis , Regulação da Expressão Gênica/efeitos dos fármacos , Interleucina-6/metabolismo , Isoflavonas/farmacologia , Camundongos , Óxido Nítrico Sintase Tipo II/metabolismo , Quercetina/farmacologia , Regulação para Cima/efeitos dos fármacos , Replicação Viral/efeitos dos fármacos
17.
Food Environ Virol ; 9(1): 35-44, 2017 03.
Artigo em Inglês | MEDLINE | ID: mdl-27807684

RESUMO

Human noroviruses (HuNoVs) cause foodborne and waterborne viral gastroenteritis worldwide. Because HuNoV culture systems have not been developed thus far, no available medicines or vaccines preventing infection with HuNoVs exist. Some herbal extracts were considered as phytomedicines because of their bioactive components. In this study, the inhibitory effects of 29 edible herbal extracts against the norovirus surrogates murine norovirus (MNV) and feline calicivirus (FCV) were examined. FCV was significantly inhibited to 86.89 ± 2.01 and 48.71 ± 7.38% by 100 µg/mL of Camellia sinensis and Ficus carica, respectively. Similarly, ribavirin at a concentration of 100 µM significantly reduced the titer of FCV by 77.69 ± 10.40%. Pleuropterus multiflorus (20 µg/mL) showed antiviral activity of 53.33 ± 5.77, and 50.00 ± 16.67% inhibition was observed after treatment with 20 µg/mL of Alnus japonica. MNV was inhibited with ribavirin by 59.22 ± 16.28% at a concentration of 100 µM. Interestingly, MNV was significantly inhibited with 150 µg/mL Inonotus obliquus and 50 µg/mL Crataegus pinnatifida by 91.67 ± 5.05 and 57.66 ± 3.36%, respectively. Treatment with 20 µg/mL Coriandrum sativum slightly reduced MNV by 45.24 ± 4.12%. The seven herbal extracts of C. sinensis, F. carica, P. multiflorus, A. japonica, I. obliquus, C. pinnatifida, and C. sativum may have the potential to control noroviruses without cytotoxicity.


Assuntos
Antivirais/farmacologia , Calicivirus Felino/efeitos dos fármacos , Norovirus/efeitos dos fármacos , Extratos Vegetais/farmacologia , Plantas/química , Animais , Antivirais/isolamento & purificação , Calicivirus Felino/crescimento & desenvolvimento , Norovirus/crescimento & desenvolvimento , Extratos Vegetais/isolamento & purificação
18.
Int J Food Microbiol ; 243: 1-8, 2017 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-27930940

RESUMO

The aim of this study was to compare the sequence of the astA gene found in 8 Korean and 11 Japanese Escherichia coli isolates. Conventional PCR was used to amplify the astA gene from the chromosomal and plasmid DNA preparation samples of each isolate using commercial DNA extraction kits. Cloning of the PCR products, sequence analysis, and pulse field gel electrophoresis (PFGE) were sequentially performed. An identical copy of astA in each isolate were found for 8 Korean and 8 Japanese E. coli strains isolated from bovine, porcine, and healthy human carriers. Among these, 1 Korean and 4 Japanese isolates carried a stop mutation at residue 16. Three Japanese outbreak strains (V199, V638, and 96-127-23) carried multiple clones of astA gene with multiple amino acids changes at residues 11, 16, 20, 23, 30, 33, and 34. Compared with the non-diarrheal isolates, clonal diversity and sequence variations of the astA gene in outbreak isolates may be associated with virulence potential of EAST1.


Assuntos
Escherichia coli Enterotoxigênica/genética , Escherichia coli Enterotoxigênica/isolamento & purificação , Enterotoxinas/genética , Proteínas de Escherichia coli/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Bovinos , Eletroforese em Gel de Campo Pulsado , Escherichia coli Enterotoxigênica/patogenicidade , Infecções por Escherichia coli/microbiologia , Fezes/microbiologia , Temperatura Alta , Humanos , Japão , Carne/microbiologia , Plasmídeos , Reação em Cadeia da Polimerase , República da Coreia , Análise de Sequência de DNA , Suínos/microbiologia , Doenças dos Suínos/microbiologia , Virulência/genética
19.
Food Environ Virol ; 9(3): 260-269, 2017 09.
Artigo em Inglês | MEDLINE | ID: mdl-28205130

RESUMO

The aim of this study was to isolate and characterize Bacillus cereus bacteriophages of various origins. Twenty-seven bacteriophages against B. cereus were isolated from various Korean traditional fermented foods and soils. Plaque size, transmission electron microscopy, virulence profile, and in vitro lytic activity of bacteriophage isolates were examined. Transmission electron microscopy confirmed B. cereus bacteriophages belonging to the family Siphoviridae. Among B. cereus bacteriophages with broad host range, 18 isolates (66.7%) did not harbor any B. cereus virulence factors. Among them, bacteriophage strain CAU150036, CAU150038, CAU150058, CAU150064, CAU150065, and CAU150066 effectively inhibited B. cereus in vitro within 1 h. Therefore, they are considered potential candidates for controlling the contamination of B. cereus in food or other applications.


Assuntos
Fagos Bacilares/isolamento & purificação , Bacillus cereus/virologia , Alimentos/virologia , Siphoviridae/isolamento & purificação , Microbiologia do Solo , Fagos Bacilares/classificação , Fagos Bacilares/genética , Fagos Bacilares/fisiologia , Especificidade de Hospedeiro , Siphoviridae/classificação , Siphoviridae/genética , Siphoviridae/fisiologia , Proteínas Virais/genética , Proteínas Virais/metabolismo
20.
Korean J Food Sci Anim Resour ; 36(2): 186-93, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27194926

RESUMO

This study aimed to inhibit Escherichia coli (E. coli) O157:H7 artificially contaminated in fresh meat using bacteriophage. Among 14 bacteriophages, the highly lytic bacteriophage BPECO19 strain was selected to inhibit E. coli O157:H7 in artificially contaminated meat samples. Bacteriophage BPECO19 significantly reduced E. coli O157:H7 bacterial load in vitro in a multiplicity of infection (MOI)-dependent manner. E. coli O157:H7 was completely inhibited only in 10 min in vitro by the treatment of 10,000 MOI BPECO19. The treatment of BPECO19 at 100,000 MOI completely reduced 5 Log CFU/cm(2) E. coli O157:H7 bacterial load in beef and pork at 4 and 8h, respectively. In chicken meat, a 4.65 log reduction of E. coli O157:H7 was observed at 4 h by 100,000 MOI. The treatment of single bacteriophage BPECO19 was an effective method to control E. coli O157:H7 in meat samples.

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