RESUMO
Viral hemorrhagic septicemia virus (VHSV) poses a significant threat to global aquaculture, prompting ongoing efforts to identify potential drug candidates for disease prevention. Coumarin derivatives have recently emerged as a promising class of compounds effective against rhabdoviruses, which severely impact the aquaculture industry. In this study, we assessed the anti-VHSV activity of umbelliferone (7-hydroxycoumarin) in fathead minnow (FHM) cells and olive flounder Paralichthys olivaceus. Umbelliferone exhibited an EC50 of 100 µg/mL by reducing cytopathic effect, with a maximum cytotoxicity of 30.9 % at 750 µg/mL. Mechanistic analyses via a time-course plaque reduction assay revealed that direct incubation with the virus for 1 h resulted in 97.0 ± 1.8 % plaque reduction, showing excellent direct virucidal activity. Pretreatment for 4 h resulted in a 33.5 ± 7.8 % plaque reduction, which increased with longer incubation times. Cotreatment led to a 33.5 ± 2.9 % plaque reduction, suggesting interference with viral binding, whereas postinfection treatment proved less effective. Umbelliferone was prophylactically administered to the olive flounder through short-term (3 days) and long-term (14 days) medicated feeding, followed by a 4-day postinfection period. Short-term administration at 100 mg/kg body weight (bw)/day resulted in the highest relative percent survival (RPS) of 56 %, whereas long-term administration achieved a maximum RPS of 44 % at 30 mg/kg bw/day. Umbelliferone administration delayed mortality at these doses. Additionally, umbelliferone significantly inhibited the expression of the VHSV N gene during viral challenge, with no observed toxic effects in fish up to an administration dose of 30 mg/kg bw/day for 28 days. Our findings suggest that the protective mechanism of short-term administration of 100 mg umbelliferone against VHSV infection may involve the overexpression of TLR2, MDA5, STAT1, and NF-κB at 24 h postinfection (hpi). IL-8 and IFN II expression was upregulated, whereas TNF-α, IL-1ß, and IFN I expression was suppressed at 24 hpi. The upregulation of ISG15 at 48 hpi may contribute to the inhibition of VHSV replication, whereas the downregulation of Caspase 3 expression at 96 hpi suggests a possible inhibition of virus-induced apoptosis at later infection stages. Overall, umbelliferone exhibited anti-VHSV activity through multiple mechanisms, with the added advantage of convenient administration via medicated feed.
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Antivirais , Novirhabdovirus , Umbeliferonas , Animais , Umbeliferonas/farmacologia , Antivirais/farmacologia , Novirhabdovirus/fisiologia , Novirhabdovirus/efeitos dos fármacos , Doenças dos Peixes/tratamento farmacológico , Doenças dos Peixes/imunologia , Doenças dos Peixes/virologia , Linguado/imunologia , Septicemia Hemorrágica Viral/virologia , Septicemia Hemorrágica Viral/imunologia , Linhagem CelularRESUMO
Viral hemorrhagic septicemia causes considerable economic losses for Korea's olive flounder (Paralichthys olivaceus) aquaculture farms; therefore, effective antiviral agents for controlling viral hemorrhagic septicemia virus (VHSV) infection are imperative. The present study implemented a Box-Behnken design and cytopathic reduction assay to derive an optimized extract of Sanguisorba officinalis L. roots (OE-SOR) with maximum antiviral activity against VHSV. OE-SOR prepared under optimized extraction conditions (55% ethanol concentration at 50 °C for 5 h) exhibited potent antiviral activity against VHSV, with a 50% effective 0.21 µg/mL concentration and a 340 selective index. OE-SOR also showed direct virucidal activity in the plaque reduction assay. Administering OE-SOR to olive flounder exhibited substantial efficacies against VHSV infection. Fish receiving 100 mg/kg body weight/day of OE-SOR as a preventive (40.0%; p < 0.05) or therapeutic (44.4%; p < 0.05) exhibited a higher relative survival than the untreated VHSV-infected control group (mortalities of 100% and 90%, respectively). In addition, fish fed with OE-SOR (100 mg/kg body weight/day) for two weeks conveyed a significantly higher inflammatory cytokine expression (nuclear factor kappa-light-chain-enhancer of activated B cells [NF-κB], interleukin-1 beta [IL-1ß], and tumor necrosis factor-alpha [TNF-α]) than the control group one to two days post-administration. Moreover, no hematological or histological changes were observed in olive flounder treated with OE-SOR over four weeks. Liquid chromatography-quadrupole-time of flight tandem mass spectrometry and -triple quadrupole tandem mass spectrometry analyses identified ziyuglycoside I as a prominent OE-SOR constituent and marker compound (content: 14.5%). This study verifies that OE-SOR is an effective alternative for controlling viral hemorrhagic septicemia in olive flounder farms as it exhibits efficient in vivo anti-VHSV activity and increases innate immune responses.
Assuntos
Doenças dos Peixes , Linguado , Septicemia Hemorrágica Viral , Novirhabdovirus , Sanguisorba , Animais , Septicemia Hemorrágica Viral/prevenção & controle , Antivirais/farmacologia , Novirhabdovirus/fisiologia , Peso Corporal , Doenças dos Peixes/prevenção & controleRESUMO
Antimicrobial peptides (AMPs) are considered a novel approach to stimulate fish antiviral mechanisms for defense against a broad range of viral infections by enhancing immunomodulatory activities. Octominin is an AMP derived from the defense proteins of Octopus minor. In this study, preliminary screening of octominin against viral hemorrhagic septicemia virus (VHSV), infectious hematopoietic necrosis virus (IHNV), and infectious pancreatic necrosis virus (IPNV) was carried out. Moreover, immune responses upon octominin treatment and IHNV challenge were investigated using fathead minnow (FHM) cells. The CC50s of octominin for FHM and Chinook salmon embryo-214 (CHSE-214) cells were 2146.2 and 1865.2 µg/mL, respectively. With octominin treatment, EC50 resulted in 732.8, 435.1, and 925.9 µg/mL for VHSV, IHNV, and IPNV, respectively. The selectivity indices were 2.9, 4.9, and 2.0, respectively. The transcriptional analysis results demonstrated the induced transcription factors (Irf3; 143-fold, Irf7; 105-fold, and NF-κB; 8-fold), stress response gene (HspB8; 2-fold), and apoptosis functional gene (p53; 3-fold) in octominin treated (500 µg/mL) FHM cells for 48 h. Moreover, IHNV viral copy number was slightly decreased with the octominin treatment (500 µg/mL) in FHM cells. Overall results suggest that octominin could be a potential antiviral agent, although further studies are necessary to understand its mode of action and the mechanism of its antiviral activity.
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Cyprinidae , Doenças dos Peixes , Vírus da Necrose Hematopoética Infecciosa , Vírus da Necrose Pancreática Infecciosa , Animais , Linhagem Celular , Peptídeos Antimicrobianos , Vírus da Necrose Pancreática Infecciosa/fisiologia , Vírus da Necrose Hematopoética Infecciosa/fisiologia , Antivirais/farmacologia , ImunidadeRESUMO
Streptococcus zoonotic bacteria cause serious problems in aquaculture with clinical effects on humans. A structure-antibacterial activity relationships analysis of 22 isoflavones isolated from M. tricuspidata (leaves, ripe fruits, and unripe fruits) against S. iniae revealed that prenylation of the isoflavone skeleton was an important key for their antibacterial activities (minimum inhibitory concentrations: 1.95-500 µg/mL). Through principal component analysis, characteristic prenylated isoflavones such as 6,8-diprenlygenistein (4) were identified as pivotal compounds that largely determine each part's antibacterial activities. M. tiricuspidata ripe fruits (MTF), which showed the highest antibacterial activity among the parts tested, were optimized for high antibacterial activity and low cytotoxicity on fathead minnow cells using Box-Behnken design. Optimized extraction conditions were deduced to be 50%/80 °C/7.5 h for ethanol concentration/extraction temperature/time, and OE-MTF showed contents of 6,8-diprenlygenistein (4), 2.09% with a MIC of 40 µg/mL. These results suggest that OE-MTF and its active isoflavones have promising potential as eco-friendly antibacterial agents against streptococcosis in aquaculture.
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Antibacterianos , Cyprinidae/microbiologia , Doenças dos Peixes , Frutas/química , Isoflavonas , Maclura/química , Extratos Vegetais/química , Streptococcus iniae/crescimento & desenvolvimento , Animais , Antibacterianos/química , Antibacterianos/isolamento & purificação , Antibacterianos/farmacologia , Doenças dos Peixes/tratamento farmacológico , Doenças dos Peixes/microbiologia , Isoflavonas/química , Isoflavonas/isolamento & purificação , Isoflavonas/farmacologia , PrenilaçãoRESUMO
An aortoesophageal fistula (AEF) is an extremely rare, potentially fatal condition, and aortic surgery is usually performed together with extracorporeal circulation. However, this surgical method has a high rate of surgical complications and mortality. This report describes an AEF caused by tuberculous esophagitis that was treated successfully using a two-stage operation. A 52-yr-old man was admitted to the hospital with severe hematemesis and syncope. Based on the computed tomography and diagnostic endoscopic findings, he was diagnosed with an AEF and initially underwent thoracic endovascular aortic repair. Esophageal reconstruction was performed after controlling the mediastinal inflammation. The patient suffered postoperative anastomotic leakage, which was treated by an endoscopic procedure, and the patient was discharged without any further problems. The patient received 9 months of anti-tuberculosis treatment after he was diagnosed with histologically confirmed tuberculous esophagitis; subsequently, he was followed as an outpatient and has had no recurrence of the tuberculosis or any further issues.
Assuntos
Doenças da Aorta/cirurgia , Esofagite Eosinofílica/complicações , Esofagite Eosinofílica/cirurgia , Fístula Esofágica/cirurgia , Esofagoscopia/métodos , Tuberculose/complicações , Doenças da Aorta/etiologia , Fístula Esofágica/etiologia , Humanos , Masculino , Pessoa de Meia-Idade , Resultado do Tratamento , Tuberculose/cirurgia , Procedimentos Cirúrgicos Vasculares/métodosRESUMO
The occurrence of an abdominal wall hematoma caused by abdominal paracentesis in patients with liver cirrhosis is rare. This paper presents a case of an abdominal wall hematoma caused by abdominal paracentesis in a 67-year-old woman with liver cirrhosis with a review of the relevant literature. Two days prior, the patient underwent abdominal paracentesis for symptom relief for refractory ascites at a local clinic. Upon admission, a physical examination revealed purpuric patches with swelling and mild tenderness in the left lower quadrant of the abdominal wall. Abdominal computed tomography revealed advanced liver cirrhosis with splenomegaly, tortuous dilatation of the para-umbilical vein, a large volume of ascites, and a large acute hematoma at the left lower quadrant of the abdominal wall. An external iliac artery angiogram showed the extravasation of contrast media from the left deep circumflex iliac artery. Embolization of the target arterial branches using N-butyl-2-cyanoacrylate was then performed, and the bleeding was stopped. The final diagnosis was an abdominal wall hematoma from the left deep circumflex iliac artery after abdominal paracentesis in a patient with liver cirrhosis.
Assuntos
Parede Abdominal , Embolização Terapêutica , Hematoma , Artéria Ilíaca , Cirrose Hepática , Paracentese , Tomografia Computadorizada por Raios X , Humanos , Feminino , Idoso , Hematoma/etiologia , Hematoma/diagnóstico , Hematoma/terapia , Cirrose Hepática/complicações , Artéria Ilíaca/diagnóstico por imagem , Angiografia , Ascite/etiologia , Ascite/terapiaRESUMO
A positive resection margin after colorectal endoscopic submucosal dissection (ESD) is associated with an increased risk of recurrence. We aimed to identify the clinical significance of positive resection margins in colorectal neoplasms after ESD. We reviewed 632 patients who had en bloc colorectal ESD at two hospitals between 2015 and 2020. The recurrence rates and presence of residual tumor after surgery were evaluated. The rate of additional surgery after ESD and recurrence rate were significantly higher in patients with incomplete resection (n = 75) compared to patients with complete resection (n = 557). When focusing solely on non-invasive lesions, no significant differences in recurrence rates were observed between the groups with complete and incomplete resection (0.2% vs. 1.9%, p = 0.057). Among 84 patients with submucosal invasive carcinoma, 39 patients underwent additional surgery due to non-curative resection. Positive vertical margin and lymphovascular invasion were associated with residual tumor. Lymphovascular invasion was associated with lymph node metastasis. However, no residual tumor nor lymph node metastases were found in patients with only one unfavorable histological factor. In conclusion, a positive resection margin in non-invasive colorectal lesions, did not significantly impact the recurrence rate. Also, in T1 colorectal cancer with a positive vertical resection margin, salvage surgery can be considered in selected patients with additional risk factors.
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Neoplasias Colorretais , Ressecção Endoscópica de Mucosa , Margens de Excisão , Recidiva Local de Neoplasia , Humanos , Neoplasias Colorretais/cirurgia , Neoplasias Colorretais/patologia , Masculino , Feminino , Ressecção Endoscópica de Mucosa/métodos , Idoso , Recidiva Local de Neoplasia/patologia , Pessoa de Meia-Idade , Neoplasia Residual/patologia , Resultado do Tratamento , Estudos Retrospectivos , Idoso de 80 Anos ou mais , Metástase LinfáticaRESUMO
Cardiovascular disease is a primary cause of mortality worldwide, and patients often require bypass surgery that utilizes autologous vessels as conduits. However, the limited availability of suitable vessels and the risk of failure and complications have driven the need for alternative solutions. Tissue-engineered vascular grafts (TEVGs) offer a promising solution to these challenges. TEVGs are artificial vascular grafts made of biomaterials and/or vascular cells that can mimic the structure and function of natural blood vessels. The ideal TEVG should possess biocompatibility, biomechanical mechanical properties, and durability for long-term success in vivo. Achieving these characteristics requires a multi-disciplinary approach involving material science, engineering, biology, and clinical translation. Recent advancements in scaffold fabrication have led to the development of TEVGs with improved functional and biomechanical properties. Innovative techniques such as electrospinning, 3D bioprinting, and multi-part microfluidic channel systems have allowed the creation of intricate and customized tubular scaffolds. Nevertheless, multiple obstacles must be overcome to apply these innovations effectively in clinical practice, including the need for standardized preclinical models and cost-effective and scalable manufacturing methods. This review highlights the fundamental approaches required to successfully fabricate functional vascular grafts and the necessary translational methodologies to advance their use in clinical practice.
Assuntos
Bioprótese , Doenças Cardiovasculares , Humanos , Prótese Vascular , Engenharia Tecidual/métodos , Materiais Biocompatíveis , Alicerces TeciduaisRESUMO
BACKGROUND/AIM: Cytochrome P450 family 46 subfamily A member 1 (CYP46A1) has been implicated in the development and progression of various cancers. This study aimed to analyze the expression of CYP46A1, examining its relationship with oncogenic behaviors, and determining its prognostic implications in colorectal cancer (CRC). MATERIALS AND METHODS: A total of 225 patients with CRC who underwent curative surgical resection were examined using paraffin-embedded tissue blocks and subjected to tumor-specific survival analysis. The expression of CYP46A1 was assessed in CRC tissues through reverse transcription-polymerase chain reaction, western blotting, and immunohistochemistry. The CRC cells' apoptosis, proliferation, angiogenesis, and lymphangiogenesis were analyzed using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assays, alongside immunohistochemical staining for Ki-67, CD34, and D2-40 antibodies. RESULTS: CYP46A1 expression was found to be up-regulated in CRC tissues compared to normal colorectal mucosa. Such expression was significantly associated with advanced stage, deeper tumor invasion, lymph node metastasis, distant metastasis, and decreased survival. Furthermore, the mean Ki-67 labeling index and microvessel density values in CYP46A1-positive tumors were significantly elevated compared to CYP46A1-negative tumors. However, there was no discernible correlation between CYP46A1 expression and either the apoptotic index or lymphatic vessel density value. CONCLUSION: CYP46A1 promotes CRC progression, specifically through the induction of tumor cell proliferation and angiogenesis. The insights provided may hold potential implications for future therapeutic interventions targeting CYP46A1.
Assuntos
Neoplasias Colorretais , Linfangiogênese , Humanos , Colesterol 24-Hidroxilase , Antígeno Ki-67 , Proliferação de Células , Neoplasias Colorretais/genéticaRESUMO
Colonic interposition is the main procedure used in esophageal reconstruction. We report a rare case of simultaneous treatment of an anastomotic site stricture and a neoplasm in the interpositioned colon. A 69-year-old female visited our outpatient clinic with symptoms of progressive dysphagia for 1 year. At the age of 30 years, the patient underwent esophagectomy with retrosternal colonic interposition because of severe esophageal burns after chemical ingestion. Upper gastrointestinal endoscopy revealed stricture at the anastomosis site and a 10-mm flat elevated high-grade dysplasia in the interpositioned colon. First, through-the-scope balloon dilatation was performed for strictures. However, stenosis was observed during the second upper gastrointestinal endoscopy session. Therefore, a second session of through-the-scope balloon dilatation was performed, and simultaneously, endoscopic submucosal dissection was also successfully performed. After 2 months of follow-up, stenosis persisted; consequently, balloon dilatation was performed. No recurrence of neoplasm was confirmed endoscopically. Through-the-scope balloon dilatation of the stricture site and simultaneous endoscopic submucosal dissection of the neoplasm in the interpositioned colon were successfully performed.
Assuntos
Neoplasias do Colo , Ressecção Endoscópica de Mucosa , Feminino , Humanos , Adulto , Idoso , Esofagectomia/efeitos adversos , Constrição Patológica , Neoplasias do Colo/diagnóstico , Neoplasias do Colo/cirurgia , Ressecção Endoscópica de Mucosa/efeitos adversosRESUMO
A 63-year-old patient was admitted with a sternal fracture and mass. On evaluation, most of the body of the sternum had been destroyed by a tumor. Radical resection of the sternum was performed and part of the major pectoral muscles adherent to the sternal tumor was also resected. The chest wall defect was reconstructed with mesh, bone cement, and a titanium rib plate system. Reconstruction with this method seemed to be an appropriate procedure to prevent instability of the chest wall.
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BACKGROUND: The major limitation of arteriovenous graft access is the high incidence of thrombotic occlusion. This study investigated the outcomes of our salvage strategy for thrombosed hemodialysis arteriovenous grafts (including surgical thrombectomy with balloon angioplasty) and evaluated the efficacy of intragraft curettage. METHODS: Salvage operations were performed for 290 thrombotic occluded arteriovenous grafts with clinical stenotic lesions from 2010 to 2018. Of these, 117 grafts received surgical thrombectomy and balloon angioplasty from 2010 to 2012 (group A), and 173 grafts received surgical thrombectomy and balloon angioplasty, with an additional salvage procedure using a curette and a graft thrombectomy catheter, from 2013 to 2018 (group B). Outcomes were described in terms of post-intervention primary patency and secondary patency rates. RESULTS: The post-intervention primary patency rates in groups A and B were 44.2% and 66.1% at 6 months and 23.0% and 38.3% at 12 months, respectively (p=0.003). The post-intervention secondary patency rates were 87.6% and 92.6% at 6 months and 79.7% and 85.0% at 12 months, respectively (p=0.623). Multivariate Cox regression analysis demonstrated that intragraft curettage was a positive predictor of post-intervention primary patency (hazard ratio, 0.700; 95% confidence interval, 0.519-0.943; p=0.019). CONCLUSION: Surgical thrombectomy and balloon angioplasty showed acceptable outcomes concerning post-intervention primary and secondary patency rates. Additionally, intragraft curettage may offer better patency to salvage thrombotic occluded arteriovenous grafts with intragraft stenosis.
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Since the first reported case of coronavirus disease 2019 (COVID-19) in December 2019, the numbers of confirmed cases and deaths have continued to increase exponentially despite multi-factorial efforts. Although various attempts have been made to improve the level of evidence for extracorporeal membrane oxygenation (ECMO) treatment over the past 10 years, most experts still hesitate to take an active position on whether to apply ECMO in COVID-19 patients. Several ECMO management guidelines have been published recently, but they reflect some important differences from the Korean medical system and aspects of real-world medical practice in Korea. We aimed to find evidence on the efficacy of ECMO for COVID-19 patients by reviewing the published literature and to propose expert recommendations by analyzing the Korean COVID-19 ECMO registry data.
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Rhus verniciflua is commonly known as a lacquer tree in Korea. The bark of R. verniciflua has been used as an immunostimulator in traditional medicine, but also causes allergic dermatitis due to urushiol derivatives. For the development of active natural resources with less toxicity, the antibacterial activity of various parts of R. verniciflua such as bark, lignum, leaves and fruit, together with chemical composition, were investigated. Among the various parts of R. verniciflua, lignum showed the most potent antibacterial activity against fish pathogenic bacteria such as Edwardsiella tarda, Vibrio anguillarum and Streptococcus iniae. Measurement of total phenolic content and flavonoid content clearly showed a high content of phenolic and flavonoids in lignum among the various parts of R. verniciflua. Further analysis showed a close correlation between antibacterial activity and phenolic content. In addition, methyl gallate and fustin, the major constituents of bark and lignum, showed antibacterial activity, which suggested phenolic constituents as active constituents. The content of urushiols, however, was highest in bark, but there was a trace amount in lignum. LC-MS-MS and PCA analysis showed good discrimination with the difference of phenolic composition in various parts of R. verniciflua. Taken together, phenolic compounds are responsible for the antibacterial activity of R. verniciflua. The lignum of R. verniciflua contains high content of phenolic compounds with less urushiols, which suggests efficient antibacterial activity with less toxicity. Therefore, the lignum of R. verniciflua is suggested as a good source for antibacterial material to use against fish bacterial diseases.
Assuntos
Antibacterianos/análise , Frutas/química , Fenóis/análise , Casca de Planta/química , Folhas de Planta/química , RhusRESUMO
Wine fermentations using rice media containing either Monascus koji or rice nuruk were performed and fermentative characteristics based on the koji type were investigated. Cultivations were performed in a 20 °C room in a 20 L bottle with the rice media that included Monascus rice koji at both 20 and 30%, or rice nuruk at 20%. After 22 days of cultivation, the ethanol yield reached 14.2-14.6% (v/v) for M. koji and 16.5% (v/v) for rice nuruk. This lower yield with use of M. koji was thought to be due to rapid cell concentration decreases in the later stage. Total amounts of organic acids and volatile compounds in fermentations using M. koji were 166-172 and 1779-1874 mg/L, respectively, being 8.7-12.9% and 46.3-54.1% higher than with use of rice nuruk. With M. koji, a high quality rice wine was produced with high levels of volatile compounds and monacolin K.
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RATIONALE: Although myocardial bridging (MB) is usually considered as benign, initial medical therapy and following surgical treatment in drug-refractory cases has been widely accepted for managing symptomatic MB. Before the patient proceeds to percutaneous or surgical intervention, however, the presence of objective ischemia in the corresponding myocardial territory should be documented. PATIENT CONCERN AND INTERVENTION: We herein report a 43-year-old male complaining of chest pain in whom cardiac CT with myocardial perfusion (cCTP) showed an MB of left anterior descending artery (LAD) with preoperative perfusion defect in corresponding myocardium and normalization of perfusion after supra-arterial myotomy. DIAGNOSIS: Myocardial bridging-induced ischemia. LESSONS: This case illustrates the potential utility of cCTP for the simultaneous assessment of MB and its hemodynamic significance for treatment planning and post-therapeutic evaluation although further research is needed to establish the clinical usefulness of this technique.
Assuntos
Ponte Miocárdica/complicações , Ponte Miocárdica/diagnóstico , Isquemia Miocárdica/etiologia , Adulto , Humanos , Masculino , Ponte Miocárdica/diagnóstico por imagem , Imagem de Perfusão do MiocárdioRESUMO
Several methods for endovascular aortic arch repair have been proposed to reduce the morbidity and mortality associated with conventional open surgery for aortic arch aneurysms. We report our experience with aortic arch aneurysm repair by a totally endovascular technique, that is, a "reversed" stent graft technique using branched stent grafts.
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Three types of Korean black raspberry wine were produced via alcoholic fermentation from juice, juice-pulp, and juice-pulp-seed, respectively. These wines were compared in terms of their anti-inflammatory activities and polyphenol contents. The total content of polyphenol compounds in wines was increased by 22.4% after supplementation with pulp and by 56.7% after supplementation with both pulp and seed. The reduction rate of NO evolution was highest in the order juice-pulp-seed wine, juice-pulp wine, and juice wine. Addition of the juice-pulp-seed wine at a level of 62.5-500 mg/L decreased the NO evolution rate by 40.5-94.2%. Eight fractions were obtained from juice-pulp-seed wine via ethyl acetate extraction and silica gel chromatography. Of these, the AF fraction, which exhibited the highest in vitro anti-inflammatory activity, exerted inhibitory effects on ear edema, writhing response, and vein membrane vascular permeability in mice. 3,4-Dihydroxybenzoic acid accounted for 37.6% of the total polyphenol content in the AF fraction.