First Report of Olive Mild Mosaic Virus in Imported Tulips (Tulipa gesneriana) in Korea.

Tulip cultivation in Korea primarily uses imported bulbs due to the absence of domestic production. To ensure safety and sustainability, Korean authorities have implemented strict phytosanitary measures for five viruses: arabis mosaic virus, tobacco necrosis virus, tobacco ringspot virus, tomato black ring virus, and tomato bushy stunt virus. In April 2021, 86 tulip plants presented symptoms such as chlorotic mottle, mosaic, streak, stripe, yellowing of the leaves, and color breaking on flowers. These samples were collected to investigate the incidence of viruses in four Korean provinces (Gangwon, Gyeongbuk, Gyeongnam, and Chungnam). The leaves and petals from each individual sample (10 mg each) were pooled and ground using liquid nitrogen. Total RNA was extracted using a Maxwell® 16 LEV Plant RNA Kit (Promega, Madison, USA). A cDNA library was constructed using TruSeq Standard Total RNA with Ribo - Zero (Illumina, San Diego, USA) and sequenced on an Illumina NovaSeq 6000 platform (Macrogen, Seoul, Korea) with 100-bp paired-end reads. Trinity software identified tulip breaking virus (TBV), tulip virus X (TVX), and lily symptomless virus (LSV), which are known to occur in Korea (Bak et al. 2023) by de novo assembly of 628 million reads into 498,795 contigs. The contigs were annotated as previously described (Bak et al. 2022). Moreover, a contig (ON758350) related to olive mild mosaic virus (OMMV; genus Alphanecrovirus, family Tombusviridae) was identified through BLASTn analysis. This contig had a 99.27% nucleotide (nt) identity to OMMV PPO-L190209 (KU641010), which was assembled from 201,346 reads and spanned 3,713 bp. To confirm the presence of OMMV, a primer pair (5'-GAATGTCTGGCGTTAAGCG-3'/5'-GTGTCCTGCGCATCATACAC-3') was designed to amplify a 797-bp fragment of the coat protein gene. In RT-PCR, 31.4% (27/86) of samples were positive for OMMV and coinfected with TBV or TBV+LSV. Coinfection with TBV led to chlorotic mottling and stripes, whereas triple coinfection with TBV+LSV produced distinct yellow streaks and mosaic within the lesion boundaries. In contrast, solely TBV infection did not produce such symptoms. The samples infected with OMMV were exclusively collected from Gangwon and Gyeongnam. In each province, an RT-PCR amplicon was cloned, and subsequently sequenced (Bioneer, Daejeon, Korea). The obtained sequences were named CC (OM243091) and GS (OM243092), and they shared 98.6% and 98.9% identity with PPO-L190209 (KU641010), respectively. A bioassay was conducted using a leaf infected with OMMV CC and TBV to inoculate 13 indicator species in triplicate, including Capsicum annuum, Chenopodium amaranticolor, C. quinoa, Cucumis sativus, Nicotiana benthamiana, N. clevelandii, N. glutinosa, N. occidentalis, N. rustica, N. tabacum, Solanum lycopersicum, Tetragonia tetragonioides, and Tulipa gesneriana. The RT-PCR revealed positivity only for OMMV in the upper leaves of N. clevelandii, while all other species were negative with no symptoms. To our knowledge, this is the first report of OMMV occurring in tulips grown from imported bulbs in Korea, with no other known natural hosts such as olive tree (Cardoso et al. 2004), spinach (Gratsia et al. 2012), and corn salad (Verdin et al. 2018). The Korean OMMV isolates exhibited a high nt identity with the foreign isolate, and the samples were collected from farms that rely entirely on bulb imports for cultivation. These suggest that the outbreak of OMMV was likely caused by imported bulbs.

Temporal changes of PIRADS scoring by radiologists and correlation to radical prostatectomy pathological outcomes.

Purpose: To assess temporal improvement of prostate image reporting and data system (PIRADS) 3-5 lesion correlation to histopathologic findings from radical prostatectomy (RP) in prostate cancer (PCa). Materials and methods: A total of 1481 patients who underwent RP for biopsy-proven PCa between 2015 and 2019 were divided into 14 groups of 100 sequential readings for the evaluation of histopathological correlation with PIRADS readings. Temporal trends of PIRADS distribution and predictive performance for RP pathology were evaluated to assess underlying changes in prostate magnetic resonance imaging (MRI) interpretation by radiologists. Results: PIRADS 4-5 lesions were significantly correlated with the increasing rates of Gleason Group (GG) upgrade (p = 0.044) and decreasing rate of GG downgrade (p = 0.016) over time. PIRADS ≥3 lesions read after median 2 years of experience were shown to independently predict intermediate-high-risk (GG ≥ 3) PCa (odds ratio 2.93, 95% confidence interval 1.00-8.54; P= 0.049) in RP pathology. Preoperative GG ≥ 3 biopsy lesions with PIRADS 4-5 lesions were significantly more susceptible to GG upgrade (P= 0.035) and GG ≥ 4 RP pathology (p = 0.003) in experienced reads, in contrast to insignificant findings in early readings (p = 0.588 and 0.248, respectively). Conclusion: Preoperative MRI reports matched with RP pathology suggest an improved prediction of adverse pathology in PIRADS 3-5 lesions over time, suggesting a temporal change in PIRADS interpretation and predictive accuracy. Institutions with low volume experience should use caution in solely relying on MRI for predicting tumor characteristics. Future prospective trials and larger scale assessments are required to further validate our results.

Pheophorbide A isolated from Gelidium amansii inhibits adipogenesis by regulating adipogenic transcription factors and AMPK in 3T3-L1 adipocytes.

Adipocyte lipid accumulation causes adipocyte hypertrophy and adipose tissue increment, leading to obesity. As part of our efforts to isolate antiobesity agents from natural products, we first isolated the active compound from the extract of Gelidium amansii through bioassay-guided fractionation. We then hypothesized that pheophorbide A isolated from G amansii inhibits adipogenesis by downregulating adipogenic transcription factors; therefore, the antiadipogenic effects of pheophorbide A were investigated in 3T3-L1 adipocytes. On differentiation of 3T3-L1 preadipocytes into adipocytes, they were treated with pheophorbide A (0-83 µM). Pheophorbide A inhibited triglyceride accumulation (half maximal inhibitory concentration = 114.2 µM) and stimulated glycerol release in a dose-dependent manner in 3T3-L1 adipocytes. In addition, pheophorbide A significantly decreased leptin concentrations in 3T3-L1 adipocytes. Pheophorbide A inhibited adipogenesis by suppressing the expression of adipogenic transcriptional factors including peroxisome proliferator-activated receptor γ, CCATT/enhancer binding protein α, sterol regulatory element binding protein 1c, and fatty acid synthase. It also induced the expression of phosphorylation of AMP-activated protein kinase. Therefore, these results suggest that pheophorbide A may be useful for preventing or treating obesity because of its inhibitory effect on adipogenesis.

First report of Tulip Virus X infecting tulip (Tulipa gesneriana) in Korea.

Tulip virus X (tulip virus X, TVX) is a member of the genus Potexvirus (family Alphaflexiviridae) and is a positive single-stranded RNA virus. TVX was described first in Scotland (Mowat 1982), followed by several countries (Yamaji et al. 2001; Tzanetakis et al. 2005; Ward et al. 2008; Dees et al. 2011; Sochacki and Komorowska 2012; Wylie et al. 2019). In April 2021, 86 whole tulip plants showing viral symptoms in leaves (mosaic, yellowing, and malformation) and flowers (color breaking) were collected in Chilgok, Chuncheon, Goseong, Yecheon and Yesan, Korea. Furthermore, high-throughput sequencing was performed to identify viruses that infect tulips in Korea. Total RNA was extracted from pooled the leaves and petals using a Maxwell® 16 LEV Plant RNA Kit (Promega, Madison, USA). We constructed a single library using the TruSeq Stranded Total RNA LT Sample Prep Kit for Plant (Illumina, San Diego, USA). The library was 100 bp paired-end sequenced using Illumina's NovaSeq 6000 (Macrogen, Seoul, Korea) and was assembled de novo using Trinity software version trinityrnaseq_r20140717, with default parameters. The contigs were annotated as in previous study (Lee et al. 2020), revealing a single contig each related to TVX, lily symptomless virus (LSV), and tulip breaking virus (TBV) was generated from 648 million total reads. The TVX-related contig (GenBank ON205948) consisting of 6,076 bp showed 99.52% nucleotide identity (6027/6056 bp) with TVX-J (GenBank AB066288). We conducted an RT-PCR assay to validate the presence of viruses with specific primers as TVX-F5093/R5624 (5'-CTATCCGGACTCATTCTACTTC/GTGCGTTCCAGATAAGCTTG-3'), LSV-F7013/R7338 (5'-CTTGGTCGACAGGGACATAAC/GATTGGAATTGTGCTTTTCAGC-3'), and TBV-F7515/R8116 (5'-GTGTGTCATGGATGATTGTTG/CAACTGATTTGCTACCGCTAG-3'). Consequently, TVX were detected in 13 of 86 samples. Moreover, LSV and TBV were detected in 15 and 26 samples, respectively. However, the yellowing and mosaic observed in the TVX infected samples were not observed in the LSV and TBV infected samples. Subsequently, two TVX amplicons were selected, cloned and sequenced. The obtained sequences were 532 bp and were named YS24 and YS38 (GenBank LC664027 and LC664028), respectively. The Korean isolates showed 98.68% (525/532 bp) and 99.62% (530/532 bp) identity with Australian isolate (GenBank MH886522) in BLASTn analysis. To bioassay for TVX, the infected tulip leaf tissue from which YS24 was obtained was used to sap-inoculate, in triplicates, 15 species of indicator plants (Nicotiana benthamiana, N. clevelandii, N. debneyi, N. glutinosa, N. rustica, N. tabacum, Datura stramonium, Glycine max, Phaseolus vulgaris, Chenopodium amaranticolor, C. quinoa, Cucumis sativus, Cu. melo, Gomphrena globosa, and Tetragonia tetragonioides). After 14 days of inoculation, we observed distinct chlorotic spots on inoculated and upper leaves of C. quinoa, but no symptoms were observed in other indicator plants. In RT-PCR assay using TVX-specific primers, only C. quinoa showed a positive reaction. In previous studies, C. amaranticolor, C. quinoa, G. globosa, and N. benthamiana were known as the experimental host of TVX (Dees et al. 2011; Tzanetakis et al. 2005), but only C. quinoa was confirmed to be susceptible to the Korean isolate. Furthermore, transmission electron microscopy revealed typical flexuous rod-shaped viral particles in the inoculated C. quinoa. To our knowledge, this is the first report of TVX infecting tulips in Korea.

Clinical Outcomes of Surgical Repair with a Composite Graft for Abdominal Aortic Aneurysm Accompanied by Iliac Artery Aneurysm.

BACKGROUND: Iliac artery aneurysm is frequently found in patients undergoing surgical repair of an abdominal aortic aneurysm. The use of commercial bifurcated grafts is insufficient for aorto-biiliac replacement with complete iliac artery aneurysm resection. We evaluated the effectiveness of handmade composite grafts for this purpose. METHODS: A total of 233 patients underwent open surgery for abdominal aortic aneurysm between 2003 and 2019, including 155 patients (67%) treated with commercial grafts and 78 patients (33%) treated with handmade composite grafts. Their operative characteristics, postoperative outcomes, and late outcomes were retrospectively reviewed. RESULTS: The early mortality rate did not differ significantly between the groups. On average, the handmade composite graft technique took approximately 15 minutes longer than the commercial graft technique (p=0.037). Among patients who underwent elective surgery, no significant differences between the conventional and composite groups were observed in the major outcomes, including red blood cell transfusion volume (2.8±4.7 units vs. 3.1±4.7 units, respectively; p=0.680), reoperation for bleeding (2.7% vs. 3.1%, respectively; p>0.999), bowel ischemia (0% vs. 1.6%, respectively; p=0.364), and intensive care unit stay duration (1.9±6.6 days vs. 1.6±2.4 days, respectively; p=0.680). The incidence of target vessel occlusion also did not differ significantly between groups. CONCLUSION: The increased technical demand involved with handmade composite grafting did not negatively impact the outcomes. This technique may be a viable option because it overcomes problems associated with commercial grafts.

The effect of 5 alpha-reductase inhibitor therapy on prostate cancer detection in the era of multi-parametric magnetic resonance imaging.

We aimed to evaluate the effect of 5 alpha-reductase inhibitor (5-ARI) treatment on prostate cancer (PCa) and clinically significant PCa (csPCa) detection in patients undergoing transrectal ultrasound (TRUS)/magnetic resonance imaging (MRI) fusion biopsy in the current era of multi-parametric MRI (mpMRI). We retrospectively reviewed our TRUS/MRI fusion biopsy database (n = 706). Eighty (11.3%) patients who had used 5-ARI for more than one year at the time of biopsy were stratified as 5-ARI group. Subsequently, we performed comparative analyses of 5-ARI and non-5-ARI groups. csPCa was defined by a Gleason score ≥3 + 4 in a single biopsy core. Chi-squared test was used to evaluate the performance of mpMRI in predicting PCa/csPCa between the two groups. Multivariate logistic regression analyses were performed to evaluate the significant variables associated with PCa detection. There were no significant differences in PCa/csPCa detection rates between 5-ARI and non-5-ARI groups (all, P > 0.05). In multivariate logistic regression analyses for the evaluation of variables associated with csPCa detection, age (odds ratio [OR], 1.062; 95% confidence interval [CI], 1.035-1.090; P < 0.001), pre-biopsy PSA (OR, 1.062; 95% CI, 1.034-1.090; P < 0.001), prostate volume on TRUS (OR, 0.956; 95% CI, 0.943-0.970, P < 0.001), and PI-RADsV2 category (OR, 5.528; 95% CI, 3.017-10.131; P < 0.001) were found to be significant predictors. However, 5-ARI had no significant association with PCa detection (P = 0.384). Conclusively, 5-ARI therapy did not adversely affect PCa/csPCa detection after TRUS/MRI fusion biopsy, which suggests that exposure to 5-ARI may not impair the performance of mpMRI.

Pheophorbide A from Gelidium amansii improves postprandial hyperglycemia in diabetic mice through α-glucosidase inhibition.

This study was designed to determine the inhibitory effects of pheophorbide A on carbohydrate digesting enzymes and its ability to improve postprandial hyperglycemia in streptozotocin (STZ)-induced diabetic mice. Pheophorbide A caused noticeable inhibitory effects on α-glucosidase and α-amylase, with half-maximal inhibitory concentrations (IC50 ) of 80.65 ± 5.90 and 76.48 ± 6.31 µM, respectively. The pheophorbide-mediated inhibition of α-glucosidase and α-amylase was significantly more effective than that of the positive control, acarbose. The increase in postprandial blood glucose levels was more significantly suppressed in the pheophorbide A group than in the control group of STZ-induced diabetic mice. In addition, the area under the curve was decreased by pheophorbide A intake in STZ-induced diabetic mice. Our results suggested that pheophorbide A may help to improve postprandial hyperglycemia by inhibiting the activity of carbohydrate digesting enzymes.

Comparison of oncological and perioperative outcomes of open, laparoscopic, and robotic nephroureterectomy approaches in patients with non-metastatic upper-tract urothelial carcinoma.

BACKGROUND: To compare the oncological and perioperative outcomes of different nephroureterectomy approaches in patients with non-metastatic upper tract urothelial carcinoma (UTUC). METHODS: We retrospectively analyzed the data of 422 patients who underwent open, laparoscopic, or robotic nephroureterectomy for non-metastatic UTUC. Perioperative and postoperative survival outcomes were compared using Kaplan-Meier analyses and Cox-proportional hazard models. RESULTS: Of the patients, 161, 137, and 124 were treated with an open, laparoscopic, and robotic approach, respectively. Laparoscopic and robotic approaches involved significantly less blood loss (p = 0.001), shorter hospital stay (p < 0.001), and longer operation time (p < 0.001) compared with the open approach. There were no significant differences in intraoperative complications (open, 8.1%; laparoscopic, 5.1%; robotic, 7.3%; p = 0.363) or early postoperative complications (open, 14.9%; laparoscopic, 14.6%; robotic, 13.7%; p = 0.880). The laparoscopic and robotic groups showed significantly less postoperative analgesic use (p = 0.015). The robotic group showed significantly longer progression-free, cancer-specific, and overall survivals than the open approach group on univariate Kaplan-Meier analysis, but surgery type was not significantly associated with survival outcomes per multivariate Cox proportional tests (all p-values > 0.05). CONCLUSION: The laparoscopic and robotic approaches yielded better perioperative outcomes, such as less intraoperative bleeding, shorter hospital stays, less analgesic usage, and non-inferior oncological outcomes, compared with the open approach. Further prospective studies are needed to compare these surgical techniques.

Differential modulation of immune response and cytokine profiles of Sargassum horneri ethanol extract in murine spleen with or without Concanavalin A stimulation.

The health benefit of brown seaweeds has been proclaimed for centuries, particularly in Asian countries. A brown seaweed Sargassum horneri has been suggested to have immune-boosting and anti-inflammatory/immune-regulatory effects, but their mechanism is still elusive. This study researches the immunological effect of 70% ethanol extract of S. horneri (SHE) on unstimulated and Con A-stimulated murine splenocytes. When treated alone, SHE had an immune stimulatory effect on CD3e+ CD4+ T-helper cells, CD3e+ CD8+ cytotoxic T cells, CD45+ CD11b+ macrophages, Ly-6C+ Ly-6 G+ granulocytes, and Ly6 G- Ly6Clow eosinophils. Furthermore, SHE enhanced wide spectrum of Th cytokines such as TNF-α, IFN-γ (Th1), IL-4, IL-5, IL-13 (Th2), and IL-6 (Th17), which also stimulated the macrophage polarizing cytokines and enhanced macrophage derived cytokine secretion. SHE in Con A (5 µg/mL) stimulated cells decreased T-helper, cytotoxic T cells, granulocytes, eosinophils, and monocytes. These results signify the potential immuno-modulatory effect of SHE which can be developed as a therapeutic agent in immuno-compromised individuals.

Minimally Invasive Redo Mitral Valve Replacement under Fibrillatory Arrest in a Patient with a Calcified Aorta and Patent Previous Bypass Grafts.

A 73-year-old woman who underwent combined bioprosthetic mitral valve replacement, tricuspid ring annuloplasty, and coronary artery bypass grafting 12 years previously visited our clinic due to aggravated dyspnea caused by structural valve deterioration of the mitral prosthesis. Because aortic or femoral artery cannulation and cross-clamping would have a high risk of stroke owing to severe calcification of the ascending aorta and ilio-femoral vessels, and because there was a risk of redo sternotomy due to the patent bypass grafts, a comprehensive approach including axillary artery cannulation, a minimally invasive right thoracotomy approach, and a clampless hypothermic fibrillatory arrest technique was used during redo mitral valve replacement.

Extra-Anatomic Ascending Aorta to Abdominal Aorta Bypass in Takayasu Arteritis Patients with Mid-Aortic Syndrome.

BACKGROUND: We evaluated the operative outcomes of an extra-anatomic bypass from the ascending aorta to the abdominal aorta in patients with type II or III Takayasu arteritis (TA) with mid-aortic syndrome. METHODS: From 1988 to 2014, 8 patients with type II (n=2) or III (n=6) TA underwent an ascending aorta to abdominal aorta bypass. The mean patient age was 43.5±12.2 years and the mean peak pressure gradient between the upper and lower extremities was 54.8±39.0 mm Hg. The median follow-up duration was 54.4 months (range, 17.8 to 177.4 months). RESULTS: There were no cases of operative mortality. The mean peak pressure gradient significantly decreased to -2.4±32.3 mm Hg (p=0.017 compared to the preoperative value). Late death occurred in 2 patients. The symptoms of upper extremity hypertension and claudication improved in all patients. The bypass grafts were patent at 47.1±58.9 months in 7 patients who underwent follow-up imaging studies. CONCLUSION: An extra-anatomic ascending aorta to abdominal aorta bypass could be an effective treatment option for severe aortic steno-occlusive disease in patients with type II or III TA, with favorable early and long-term outcomes.

Indentation in the Right Ventricle by an Incomplete Pericardium on 3-Dimensional Reconstructed Computed Tomography.

We report the case of a 17-year-old girl who presented with an indentation in the right ventricle caused by an incomplete pericardium on preoperative 3-dimensional reconstructed computed tomography. She was to undergo surgery for a partial atrioventricular septal defect and secundum atrial septal defect. Preoperative electrocardiography revealed occasional premature ventricular beats. We found the absence of the left side of the pericardium intraoperatively, and this absence caused strangulation of the diaphragmatic surface of the right ventricle. After correcting the lesion, the patient's rhythm disturbances improved.

Gelidium amansii extract ameliorates obesity by down-regulating adipogenic transcription factors in diet-induced obese mice.

BACKGROUND/OBJECTIVES: In this study, we investigated whether Gelidium amansii extract (GAE) ameliorates obesity in diet-induced obese (DIO) mice. MATERIALS/METHODS: The mice were maintained on a high-fat diet (HD) for 5 weeks to generate the DIO mouse model. And then mice fed HD plus 0.5% (GAE1), 1% (GAE2) or 2% (GAE3) for 8 weeks. RESULTS: After the experimental period, GAE-supplemented groups were significantly lower than the HD group in body weight gain and liver weight. GAE supplemented groups were significantly lower than the HD group in both epididymal and mesenteric adipose tissue mass. The plasma leptin level was significantly higher in the HD group than in GAE-supplemented groups. The leptin level of HD+GAE3 group was significantly lower than that of the HD+conjugated linoleic acid (CLA) group. In contrast, plasma adiponectin level of the HD group was significantly lower than those of HD+GAE2 and HD+GAE3 groups. The expression levels of adipogenic proteins such as fatty acid synthase, sterol regulatory element-binding protein-1c, peroxisome proliferator-activated receptor γ, and CCAAT/enhancer binding protein α in the GAE supplemented groups were significantly decreased than those in HD group, respectively. In addition, the expression levels of HD+GAE2 and HD+GAE3 groups are significantly decreased compared to those of HD+CLA group. On the contrary, the expression levels of hormone-sensitive lipase and phospho-AMP-activated protein kinase, proteins associated with lipolysis, were significantly increased in the GAE supplemented groups compared to those in the HD group. HD+GAE3 group showed the highest level among the GAE supplemented groups. CONCLUSIONS: These results suggested that GAE supplementation stimulated the expressions of lipid metabolic factors and reduced weight gain in HD-fed C57BL/6J obese mice.

Gelidium amansii ethanol extract suppresses fat accumulation by down-regulating adipogenic transcription factors in ob/ob mice model.

The purpose of this study was to determine the anti-obesity effects of Gelidium amansii extract (GAE) in the C57BL/6J-ob/ob mice. The ob/ob mice were fed GAE at 0.5% for 4 weeks, after which body weight, epididymal adipose tissue weight, plasma triglycerides, and hepatic lipid accumulation were significantly reduced in GAE-fed mice compared with ob/ob control mice. Plasma adiponectin levels were significantly higher in GAE-fed mice than in ob/ob control mice. These findings were supported by the expression levels of enzymes and proteins related to lipid metabolism assessed by western blotting: protein expression levels of the peroxisome proliferator-activated receptor γ and CCATT/enhancer binding protein α decreased significantly, while hormone-sensitive lipase and phospho-AMP-activated protein kinase levels increased in the GAE-fed mice compared with ob/ob control mice. These findings demonstrate that GAE regulates plasma lipid profiles and increasing highdensity lipoprotein cholesterol levels as well as by regulating the expression levels of lipid metabolic factors, resulting in reduced weight gain in ob/ob mice.

Cardiac Resynchronization Therapy Defibrillator Treatment in a Child with Heart Failure and Ventricular Arrhythmia.

Cardiac resynchronization therapy (CRT) is a new treatment for refractory heart failure. However, most patients with heart failure treated with CRT are adults, middle-aged or older with idiopathic or ischemic dilated cardiomyopathy. We treated a 12-year-old boy, who was transferred after cardiac arrest, with dilated cardiomyopathy, left bundle-branch block, and ventricular tachycardia. We performed cardiac resynchronization therapy with a defibrillator (CRT-D). After CRT-D, left ventricular ejection fraction improved from 22% to 44% assessed by echocardiogram 1 year postoperatively. On electrocardiogram, QRS duration was shortened from 206 to 144 ms. The patient's clinical symptoms also improved. For pediatric patients with refractory heart failure and ventricular arrhythmia, CRT-D could be indicated as an effective therapeutic option.

Anti-inflammatory effects of calcium citrate in RAW 264.7cells via suppression of NF-κB activation.

Here we aimed to investigate the anti-inflammatory effects of calcium citrate in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. The anti-inflammatory effects of calcium citrate were investigated by assessing pro-inflammatory factors (NO, ROS, NF-κB, iNOS, and COX-2) and pro-inflammatory cytokines (IL-1ß, IL-6, and TNF-α). Treatment of cells with calcium citrate (10-100µM) significantly reduced the generation of intracellular reactive oxygen species and increased the activities of the antioxidant enzymes superoxide dismutase, catalase, and glutathione peroxidase in LPS-stimulated macrophages. Calcium citrate was further shown to inhibit NO production in LPS-stimulated RAW 264.7cells. The expression levels of iNOS, COX-2, and NF-κB were also suppressed by treatment with calcium citrate. Calcium citrate was furthermore found to significantly inhibit the production of IL-1ß, IL-6, and TNF-α in response to LPS-stimulation. These findings demonstrate that calcium citrate may be an effective anti-inflammatory agent.

Caffeic acid phenethyl ester promotes anti-inflammatory effects by inhibiting MAPK and NF-κB signaling in activated HMC-1 human mast cells.

CONTEXT: Caffeic acid phenethyl ester (CAPE), an active component of honeybee propolis, is known to have antioxidant, anti-inflammatory, and other beneficial medicinal properties. However, the molecular mechanisms underlying its anti-allergic effects in mast cells are unknown. OBJECTIVE: The purpose of the present study was to examine whether CAPE modulates the immunoglobulin E (IgE)-mediated local allergic reaction in animals, as well as to elucidate the effects of CAPE on mast cells in vitro. MATERIALS AND METHODS: To investigate the bioactive potential of CAPE (10 or 20 µM), HMC-1 cells were stimulated with phorbol 12-myristate 13-acetate plus calcium ionophore A23187 (PMACI) for 24 h in the presence or absence of CAPE. To study the pharmacological effects of CAPE, enzyme-linked immunosorbent assays (ELISAs), RT-PCR, Western blot analysis, electrophoretic mobility shift assays (EMSAs), and fluorescence assays were used. RESULTS: CAPE (10 mg/kg) inhibited local IgE-mediated allergic reactions (0.164 versus 0.065 O.D.) in a mouse model. Additionally, CAPE (20 µM) attenuated PMACI-stimulated histamine release (3146.42 versus 2564.83 pg/ml) and the production of inflammatory cytokines, such as interleukin (IL)-1ß (4.775 versus 0.713 pg/ml, IC50 = 6.67 µM), IL-6 (4771.5 versus 449.1 pg/ml, IC50 = 5.25 µM), and IL-8 (5991.7 versus 2213.1 pg/ml, IC50 = 9.95 µM) in HMC-1 cells. In activated HMC-1 cells, pretreatment with CAPE decreased the phosphorylation of c-Jun N-terminal kinase. In addition, CAPE inhibited PMACI-induced nuclear factor (NF)-κB activation by suppressing IκBα phosphorylation and its degradation. DISCUSSION AND CONCLUSION: Our results indicated that CAPE can modulate mast cell-mediated allergic disease.

Inhibitory effect of Gastrodia elata extract on melanogenesis in HM3KO melanoma cells.

Effective and safe antimelanogenic agents derived from natural products get interest continuously for both medical and cosmetic purposes. Melanin synthesis is regulated by melanogenic enzymes such as tyrosinase, tyrosinase-related protein 1 (TRP-1), and tyrosinase-related protein 2 (TRP-2) in mammals. Gastrodia elata (GE) has been shown to have multiple therapeutic actions related to antioxidation in many diseases. In this study, we investigate whether water extraction of GE has inhibitory effects on melanogenesis in vitro and on the expression of mRNA and protein of tyrosinase, TRP-1, and TRP-2 in HM3KO melanoma cells. To examine the inhibitory effect of GE on melanogenesis, mushroom tyrosinase inhibition assay; reverse transcription polymerase chain reaction; and western blotting of tyrosinase, TRP-1, and TRP-2 were performed using HM3KO melanoma cells. In this study, the GE extract was found to significantly inhibit mushroom tyrosinase activity (69.3 ± 7.2% of the control, p < 0.05), and the expression of mRNA and protein of tyrosinase, TRP-1, and TRP-2 was reduced significantly. These results suggest that the antimelanogenic effect of GE extract is mainly due to the decreased expression of mRNA and protein of tyrosinase, TRP-1, and TRP-2 in the process of melanin synthesis.

Effect of diphlorethohydroxycarmalol isolated from Ishige okamurae on apoptosis in 3 T3-L1 preadipocytes.

Obesity is an important issue in the world of public health and preventive medicine. Inhibition of proliferation of preadipocytes plays an important role in proposed antiobesity mechanisms. In this study, we investigated the effect of diphlorethohydroxycarmalol (DPHC) isolated from Ishige okamurae on the apoptotic pathway. The results showed that DPHC inhibited population growth in 3 T3-L1 preadipocytes as assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Flow cytometric analysis of 3 T3-L1 preadipocytes showed that the number of early and late apoptotic cells increases in a dose-dependent manner after exposure to DPHC, while the number of normal cells was reduced. Our findings indicate that the induction of apoptosis in 3 T3-L1 preadipocytes by DPHC is mediated through the activation of caspase-3, Bax, and caspase-9, and then through the cleavage of poly(ADP-ribose) polymerase and the down-regulation of Bcl-2. The data also indicated that treatment with DPHC inhibits histone deacetylase activity in 3 T3-L1 preadipocytes. These results show that DPHC efficiently induces apoptosis in 3 T3-L1 preadipocytes.

Anti-inflammatory Effect of Oyster Shell Extract in LPS-stimulated Raw 264.7 Cells.

This study was designed to investigate the anti-inflammatory effect of oyster shell extract on the production of pro-inflammatory factors [NO, reactive oxygen species (ROS), nuclear factor-kappa B (NF-κB), inducible nitric oxide synthase (iNOS) and cycloxygenase-2 (COX-2)] and pro-inflammatory cytokines [Interleukin-1ß (IL-1ß), Interleukin-6 (IL-6) and TNF-α] in the lipopolysaccharide (LPS)-stimulated Raw 264.7 cells. Cell viability, as measured by the MTT assay, showed that oyster shell extract had no significant cytotoxicity in Raw 264.7 cells. The treatment with oyster shell extract decreased the generation of intracellular reactive oxygen species dose dependently and increased antioxidant enzyme activities, such as SOD, catalase, GSH-px in LPS-stimulated macrophage cells. Oyster shell extract significantly suppressed the production of NO and also decreased the expressions of iNOS, COX-2 and NF-κB. Additionally, oyster shell extract significantly inhibited the production of IL-1ß, IL-6, and TNF-α in LPS-stimulated Raw 264.7 cells. Thus, these results showed that the oyster shell extract had an anti-inflammatory effect on LPS-stimulated Raw 264.7 cells.