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EGFR is frequently amplified, mutated, and overexpressed in malignant gliomas. Yet the EGFR-targeted therapies have thus far produced only marginal clinical responses, and the underlying mechanism remains poorly understood. Using an inducible oncogenic EGFR-driven glioma mouse model system, our current study reveals that a small population of glioma cells can evade therapy-initiated apoptosis and potentiate relapse development by adopting a mesenchymal-like phenotypic state that no longer depends on oncogenic EGFR signaling. Transcriptome analyses of proximal and distal treatment responses identified TGFβ/YAP/Slug signaling cascade activation as a major regulatory mechanism that promotes therapy-induced glioma mesenchymal lineage transdifferentiation. Following anti-EGFR treatment, TGFβ secreted from stressed glioma cells acted to promote YAP nuclear translocation that stimulated upregulation of the pro-mesenchymal transcriptional factor SLUG and subsequent glioma lineage transdifferentiation toward a stable therapy-refractory state. Blockade of this adaptive response through suppression of TGFβ-mediated YAP activation significantly delayed anti-EGFR relapse and prolonged animal survival. Together, our findings shed new insight into EGFR-targeted therapy resistance and suggest that combinatorial therapies of targeting both EGFR and mechanisms underlying glioma lineage transdifferentiation could ultimately lead to deeper and more durable responses. Significance: This study demonstrates that molecular reprogramming and lineage transdifferentiation underlie anti-EGFR therapy resistance and are clinically relevant to the development of new combinatorial targeting strategies against malignant gliomas with aberrant EGFR signaling.

Liver regeneration is a very complex process and is regulated by several cytokines and growth factors. It is also known that liver transplantation and the regeneration process cause massive oxidative stress, which interferes with liver regeneration. The placenta is known to contain various physiologically active ingredients such as cytokines, growth factors, and amino acids. In particular, human placenta hydrolysate (hPH) has been found to contain many amino acids. Most of the growth factors found in the placenta are known to be closely related to liver regeneration. Therefore, in this study, we investigated whether hPH is effective in promoting liver regeneration in rats undergoing partial hepatectomy. We confirmed that cell proliferation was significantly increased in HepG2 and human primary cells. Hepatocyte proliferation was also promoted in partial hepatectomized rats by hPH treatment. hPH increased liver regeneration rate, double nucleic cell ratio, mitotic cell ratio, proliferating cell nuclear antigen (PCNA), and Ki-67 positive cells in vivo as well as interleukin (IL)-6, tumor necrosis factor alpha (TNF-α), and hepatocyte growth factor (HGF). Moreover, Kupffer cells secreting IL-6 and TNF-α were activated by hPH treatment. In addition, hPH reduced thiobarbituric acid reactive substances (TBARs) and significantly increased glutathione (GSH), glutathione peroxidase (GPx), and superoxide dismutase (SOD). Taken together, these results suggest that hPH promotes liver regeneration by activating cytokines and growth factors associated with liver regeneration and eliminating oxidative stress.

Diarrhea is a fatal disease to neonatal calves, and rotavirus is the main pathogen associated with neonatal calf diarrhea. Although previous studies have reported that the gut microbiota is changed in calves during diarrhea, less is known about whether rotavirus infection alters the structure of the gut microbiota. Here, we characterized fecal microbial communities and identified possible relationships between the gut microbiota profiles and physiological parameters. Five fecal specimens of rotavirus-infected calves from 1 to 30 days after birth and five fecal specimens of age-matched healthy calves were used for the microbial community analysis using the Illumina MiSeq sequencer. Rotavirus infection was associated with reduced rotavirus infection significantly reduced the richness and diversity of the bacterial community. Weighted unique fraction metric analysis exhibited significant differences in community membership and structure between healthy and rotavirus-infected calves. Based on relative abundance analysis and linear discriminant analysis effect size, we found that the representative genera from Lactobacillus, Subdoligranulum, Blautia, and Bacteroides were closely related to healthy calves, while the genera Escherichia and Clostridium were closely affiliated to rotavirus-infected calves. Furthermore, canonical correlation analysis and Pearson correlation coefficient results revealed that the increased relative abundances of Lactobacillus, Subdoligranulum, and Bacteroides were correlated with normal levels of physiological characteristics such as white blood cells, blood urea nitrogen, serum amyloid protein A, and glucose concentration in serum. These results suggest that rotavirus infection alters the structure of the gut microbiota, correlating changes in physiological parameters. This study provides new information on the relationship between gut microbiota and the physiological parameters of rotavirus-mediated diarrheic calves.

Amyloid beta (Aβ) aggregates are an important therapeutic target for Alzheimer's disease (AD), a fatal neurodegenerative disease. To date, AD still remains a big challenge due to no effective treatments. Based on the property that Aβ aggregates have the cross-β-structure, a common structural feature in amyloids, we systemically designed the Aβ-aggregation inhibitor that maintains Aβ-interacting ability but removes toxic part from SOD1 (superoxide dismutase 1)-G93A. We identified NABi (Natural Aβ Binder and Aβ-aggregation inhibitor) composed of β2-3 strands, a novel breaker of Aβ aggregation, which does not self-aggregate and has no cytotoxicity at all. The NABi blocks Aβ-fibril formation in vitro and in vivo and prevents neuronal cell death, a hallmark of AD pathogenesis. Such anti-amyloidogenic properties can provide novel strategies for treating AD. Furthermore, our study provides molecular insights into the design of amyloidogenic inhibitors to cure various neurodegenerative and amyloid-associated diseases, as NABi would regulate aggregation of other toxic β-sheet proteins other than Aβ.

Mantle cell lymphoma (MCL) often has an adverse prognosis and despite aggressive multimodal treatment with conventional and targeted therapies, the median survival of MCL patients remains approximately 4 years. Thus, there is a significant unmet need to find novel targets and rational combination treatments. Targeting lineage vulnerabilities driven by specific transcription factors has been broadly confirmed as an effective intervention in many human cancers. To identify the transcription program dependency of MCL cells, we conducted an unbiased domain-focused CRISPR-Cas9 screening against a library of 8,750 sgRNAs targeting 1,434 transcription factors in MCL cell lines (JEKO1, MAVER1, UPN1, and CCMCL1). We identified Forkhead Box O1 (FOXO1), EBF1, PAX5, and IRF4 as 4 transcription factors that are specifically required for MCL survival and growth. Chromatin-immunoprecipitation and sequencing (ChIP-Seq) analysis further revealed that the four transcription factors act together to orchestrate B cell lineage transcriptional program and MCL cell survival. Despite its well-recognized role as a tumor suppressor, FOXO1 has been implicated as a lineage specific transcription factor involved in mature B cell development. Genetic studies revealed a critical role of FOXO1 in germinal center dark zone formation and lymphomagenesis, raising the possibility that FOXO1 acts as a master transcription factor for lineage survival transcription program of MCL. Indeed, hierarchical interaction analysis revealed that FOXO1 functions as a pioneer factor that facilitate the chromatin access of other B cell lineage transcription factors. We demonstrated that interaction of FOXO1 to its cognate motif stabilizes B cell transcription factor complex and supports MCL progression. Along this line, we show that enforced expression of FOXO1 in myeloid leukemia cells induces transdifferentiation and B-cell specific gene expression. Mechanistically, we demonstrate through tiling CRISPR scanning screen that forkhead DNA binding and c-terminal transactivation domains of FOXO1 are specifically required for the viability of MCL cells. Given our finding of FOXO1 as a lineage-specific oncogene in MCL, we next explored the possibility of developing FOXO1-tageted inhibitors. We screened a library of potential small molecule inhibitors of FOXO1 (Forkhead BioTherapeutics) and identified cpd10 as one of the most potent and selective FOXO1 inhibitors (IC 50=76 ×/÷ 1.7. nM) Through the CCMCL1 MCL-NSG preclinical model, we found that cpd10 (100 mg/kg/daily) was well tolerated without overt toxicities. Importantly, prolonged treatment induced a robust cytotoxic response of MCL cells and suppressed MCL progression in vivo. Altogether, our findings identified FOXO1 as a MCL lineage-survival oncogene that can be exploited as a therapeutic target of future drug development. Disclosures Cantley: Petra Pharmaceuticals: Research Funding; Agios Pharmaceuticals: Membership on an entity's Board of Directors or advisory committees. Elemento: Freenome: Consultancy, Other: Current equity holder in a privately-held company; Volastra Therapeutics: Consultancy, Other: Current equity holder, Research Funding; Champions Oncology: Consultancy; Janssen: Research Funding; Owkin: Consultancy, Other: Current equity holder; AstraZeneca: Research Funding; One Three Biotech: Consultancy, Other: Current equity holder; Eli Lilly: Research Funding; Johnson and Johnson: Research Funding. Baiocchi: Prelude Therapeutics: Consultancy; viracta: Consultancy, Current holder of stock options in a privately-held company; Codiak Biosciences: Research Funding; Atara Biotherapeutics: Consultancy. Belvedere: Forkhead BioTherapeutics: Current Employment. Paik: Forkhead BioTherapeutics: Research Funding.

A novel halophilic bacterium, strain K7(T), was isolated from kimchi, a traditional Korean fermented food. The strain is Gram-positive, motile, and produces terminal endospores. The isolate is facultative aerobic and grows at salinities of 0.0-25.0% (w/v) NaCl (optimum 10-15% NaCl), pH 5.5-8.5 (optimum pH 7.0-7.5), and 15-42°C (optimum 37°C). The predominant isoprenoid quinone in the strain is menaquinone-7 and the peptidoglycan of the strain is meso-diaminopimelic acid. The major fatty acids of the strain are anteisio-C15:0, iso-C15:0, and, C16:0 (other components were < 10.0%), while the major polar lipids are diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, and three unidentified lipids. A phylogenetic analysis of 16S rRNA gene sequence similarity showed that the isolated strain was a cluster of the genus Gracilibacillus. High levels of gene sequence similarity were observed between strain K7(T) and Gracilibacillus orientalis XH-63(T) (96.5%), and between the present strain and Gracilibacillus xinjiangensis (96.5%). The DNA G+C content of this strain is 37.7 mol%. Based on these findings, strain K7(T) is proposed as a novel species: Gracilibacillus kimchii sp. nov. The type strain is K7(T) (KACC 18669(T); JCM 31344(T)).

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by progressive degeneration of motor neurons. Aggregation of ALS-linked mutant Cu/Zn superoxide dismutase (SOD1) is a hallmark of a subset of familial ALS (fALS). Recently, intracellular amyloid-β (Aβ) is detected in motor neurons of both sporadic and familial ALS. We have previously shown that intracellular Aβ specifically interacts with G93A, an ALS-linked SOD1 mutant. However, little is known about the pathological and biological effect of this interaction in neurons. In this study, we have demonstrated that the Aβ-binding region is exposed on the SOD1 surface through the conformational changes due to misfolding of SOD1. Interestingly, we found that the intracellular aggregation of Aβ is enhanced through the direct interaction of Aβ with the Aβ-binding region exposed to misfolded SOD1. Ultimately, increased Aβ aggregation by this interaction promotes neuronal cell death. Consistent with this result, Aβ aggregates was three-fold higher in the brains of G93A transgenic mice than those of non Tg. Our study provides the first direct evidence that Aβ, an AD-linked factor, is associated to the pathogenesis of ALS and provides molecular clues to understand common aggregation mechanisms in the pathogenesis of neurodegenerative diseases. Furthermore, it will provide new insights into the development of therapeutic approaches for ALS.

Herbicidin A is a potent herbicide against dicotyledonous plants as well as an antibiotic against phytopathogens. In this study, fermentation parameters for herbicidin A production in submerged culture of Streptomyces scopuliridis M40 were investigated. The herbicidin A concentration varied with the C/N ratio. High C/N ratios (>4) resulted in a herbicidin A production of more than 900 mg/l, whereas maximally 600 mg/l was obtained at ratios between 1 and 3.5. In 5-L batch fermentation, there was a positive correlation between the oxygen uptake rate (OUR) and herbicidin A production. Once the OUR increased, the substrate consumption rate increased, leading to an increase in volumetric productivity. Mechanical shear force affected the hyphal morphology and OUR. When the medium value of hyphal size ranged from 150 to 180 μm, high volumetric production of herbicidin A was obtained with OUR values >137 mg O2/l·h. The highest herbicidin A concentration of 956.6 mg/l was obtained at 500 rpm, and coincided with the highest relative abundance of hyphae of 100-200 μm length and the highest OUR during cultivation. Based on a constant impeller tip speed, which affects hyphal morphology, herbicidin A production was successfully scaled up from a 5-L jar to a 500-L pilot vessel.

Argon plasma jet (Ar-PJ) has been widely used in clinical medicine; however, the cellular effects of Ar-PJ therapy applying to living tissues have not been clarified yet. It is necessary to investigate cellular responses to Ar-PJ in establishing guidelines on the therapeutic use of Ar-PJ. Interestingly, in the Ar-PJ-treated cells, the fragmented mitochondria, a typical cellular stress indicator, were discovered even in the cells located in the live zones (1∼3 zones). Using microscopic measurements of the mitochondrial length, we found that the fragmented mitochondria were mainly in the zones 1 and 2, the closest to the direct exposure point of Ar-PJ. Whereas, the mitochondria in the zone 4 retained their lengths to normal. This quantitative measurement of mitochondrial morphology was combined with the color scores of the polymerizable supramolecular (PS) sensor in diagnostic categories. The results demonstrate that the mitochondrial length (0.98∼3.94 μm) is inversely proportional to the PS sensor color scores (87∼0) in the zones 1∼4. On the combination of these three diagnostic parameters, the effective range of Ar-PJ for cellular responses was determined: the zones 1∼3, the color scores 87∼12 and the mitochondrial lengths 0.98∼2.57 μm. Our study is the first demonstration of mitochondrial fragmentation in response to Ar-PJ and the first attempt to establish the diagnostic guideline for Ar-PJ therapies by combinations with biological, physical and chemical aspects. Thus, this study will make great advances in the field of bioplasma applications.

Anti-atherosclerosis effects of perilla oil were investigated, in comparison with lovastatin, in rabbits fed a high-cholesterol diet (HCD). Hypercholesterolemia was induced in rabbits by feeding the HCD containing 0.5% cholesterol and 1% corn oil, and perilla oil (0.1 or 0.3%) was added to the diet containing 0.5% cholesterol for 10 weeks. HCD greatly increased blood total cholesterol and low-density lipoproteins, and caused thick atheromatous plaques, covering 74% of the aortic wall. Hyper-cholesterolemia also induced lipid accumulation in the liver and kidneys, leading to lipid peroxidation. Perilla oil not only attenuated hypercholesterolemia and atheroma formation, but also reduced fat accumulation and lipid peroxidation in hepatic and renal tissues. The results indicate that perilla oil prevents atherosclerosis and fatty liver by controlling lipid metabolism, and that it could be the first choice oil to improve diet-induced metabolic syndrome.

The aim of this study is to investigate the potential of anti-osteoarthritis effects on egg white-chalcanthite (EC), purple bamboo salts (PBS), and a mixture of EC and PBS (EC+PBS). EC is a mixture of egg white and pulverized chalcanthite. PBS has been widely used as one of functional foods in Korea and shows unique features compared with common salt. Osteoarthritis was induced by intra-articular injection of monosodium iodoacetate (MIA, 4mg/kg bw) in Sprague-Dawley (SD) rats. Test substances were administered once daily for 6 weeks at doses of 10 mg EC, EC+100 mg PBS, EC+200 mg PBS before and after MIA injection. Each substance was assessed by blood chemistry parameters, and by serum cytokines including IL-1β and IL-6, and nitric oxide (NO) and prostaglandin-E2 (PGE2). Structural changes of articular cartilage were also evaluated by histopathological examination. As a result, body weight and blood chemistry parameter were not different in all experimental groups. EC+PBS mixture reduced the production of PGE2, NO, IL-1β, and IL-6. In histological grade of osteoarthritis, EC+PBS mixture had a tendency to ameliorate damage of articular cartilage induced by MIA in a dose-dependent manner. In conclusion, EC+PBS mixture was demonstrated to have a potential for anti-inflammatory effect against osteoarthritis induced by MIA in a dose-dependent manner.

A Gram-stain-positive, rod-shaped, alkalitolerant, and halophilic bacterium–designated as strain NKC3-5T–was isolated from kimchi that was collected from the Geumsan area in the Republic of Korea. Cells of isolated strain NKC3-5T were 0.5–0.7 μm wide and 1.4–2.8 μm long. The strain NKC3-5T could grow at up to 20.0% (w/v) NaCl (optimum 10%), pH 6.5–10.0 (optimum pH 9.0), and 25–40°C (optimum 35°C). The cells were able to reduce nitrate under aerobic conditions, which is the first report in the genus Salicibibacter. The genome size and genomic G + C content of strain NKC3-5T were 3,754,174 bp and 45.9 mol%, respectively; it contained 3,630 coding sequences, 16S rRNA genes (six 16S, five 5S, and five 23S), and 59 tRNA genes. Phylogenetic analysis based on 16S rRNA showed that strain NKC3-5T clustered with bacterium Salicibibacter kimchii NKC1-1T, with a similarity of 96.2–97.6%, but formed a distinct branch with other published species of the family Bacillaceae. In addition, OrthoANI value between strain NKC3-5T and Salicibibacter kimchii NKC1-1T was far lower than the species demarcation threshold. Using functional genome annotation, the result found that carbohydrate, amino acid, and vitamin metabolism related genes were highly distributed in the genome of strain NKC3-5T. Comparative genomic analysis revealed that strain NKC3-5T had 716 pan-genome orthologous groups (POGs), dominated with carbohydrate metabolism. Phylogenomic analysis based on the concatenated core POGs revealed that strain NKC3-5T was closely related to Salicibibacter kimchii. The predominant polar lipids were phosphatidylglycerol and two unidentified lipids. Anteiso-C15:0, iso-C17:0, anteiso-C17:0, and iso-C15:0 were the major cellular fatty acids, and menaquinone-7 was the major isoprenoid quinone present in strain NKC3-5T. Cell wall peptidoglycan analysis of strain NKC3-5T showed that meso-diaminopimelic acid was the diagnostic diamino acid. The phephenotypic, genomic, phylogenetic, and chemotaxonomic properties reveal that the strain represents a novel species of the genus Salicibibacter, for which the name Salicibibacter halophilus sp. nov. is proposed, with the type strain NKC3-5T (= KACC 21230T = JCM 33437T).

Liver regeneration is a very complex process and is regulated by several cytokines and growth factors. It is also known that liver transplantation and the regeneration process cause massive oxidative stress, which interferes with liver regeneration. The placenta is known to contain various physiologically active ingredients such as cytokines, growth factors, and amino acids. In particular, human placenta hydrolysate (hPH) has been found to contain many amino acids. Most of the growth factors found in the placenta are known to be closely related to liver regeneration. Therefore, in this study, we investigated whether hPH is effective in promoting liver regeneration in rats undergoing partial hepatectomy. We confirmed that cell proliferation was significantly increased in HepG2 and human primary cells. Hepatocyte proliferation was also promoted in partial hepatectomized rats by hPH treatment. hPH increased liver regeneration rate, double nucleic cell ratio, mitotic cell ratio, proliferating cell nuclear antigen (PCNA), and K

Food-grade cryoprotective agents, i.e., skim milk, soy powder, yeast extract, and trehalose were tested for viability of lactic acid bacteria (LAB, Lactobacillus brevis WK12 and Lactococcus lactis WK11) during freeze drying. Ten percentage of soy powder showed a strong protective effect upon viability of both LAB, showing 1.85×1011 CFU/mL for L. brevis WK12 and 1.89×1011 CFU/mL for L. lactis WK11. Microencapsulation in Ca-alginate beads increased viability of LAB, compared to nonencapsulated cells. Moreover, there was a synergetic effect on the stability of LAB when encapsulated LAB in Ca-alginate beads were soaked in 10% of soy powder prior to freeze-drying. When temperature increased, however, survival of LAB decreased. Four weeks after storage the highest viability was observed at -18°C for L. brevis WK12 and L. lactis WK11, showing 1.80×1011 CFU/mL and 1.78×1011 CFU/mL, respectively. These results suggest that soy powder may be useful for enhancing viability of LAB with microencapsulation.

Argon plasma jet (Ar-PJ) has been widely used in clinical medicine; however, the cellular effects of Ar-PJ therapy applying to living tissues have not been clarified yet. It is necessary to investigate cellular responses to Ar-PJ in establishing guidelines on the therapeutic use of Ar-PJ. Interestingly, in the Ar-PJ-treated cells, the fragmented mitochondria, a typical cellular stress indicator, were discovered even in the cells located in the live zones (1∼3 zones). Using microscopic measurements of the mitochondrial length, we found that the fragmented mitochondria were mainly in the zones 1 and 2, the closest to the direct exposure point of Ar-PJ. Whereas, the mitochondria in the zone 4 retained their lengths to normal. This quantitative measurement of mitochondrial morphology was combined with the color scores of the polymerizable supramolecular (PS) sensor in diagnostic categories. The results demonstrate that the mitochondrial length (0.98∼3.94 μm) is inversely proportional to the PS sensor color scores (87∼0) in the zones 1∼4. On the combination of these three diagnostic parameters, the effective range of Ar-PJ for cellular responses was determined: the zones 1∼3, the color scores 87∼12 and the mitochondrial lengths 0.98∼2.57 μm. Our study is the first demonstration of mitochondrial fragmentation in response to Ar-PJ and the first attempt to establish the diagnostic guideline for Ar-PJ therapies by combinations with biological, physical and chemical aspects. Thus, this study will make great advances in the field of bioplasma applications.

Institute of Life Sciences & BiotechnologyAbstract Cynanchum wilfordii and Phlomis umbrosa Turcz. are known to contain isoflavone, a representative phytoestrogen.This study was performed to determine whether the extract mixture of C. wilfordii and P. umbrosa Turcz. would inducean estrogenic effect in ovariectomized rats. The extracts were administered to the ovariectomized rats at 30, 60, 120 mg/kg per day for 4 weeks, respectively. They showed no estrogenic effect, which was indicated by the decrease in uteruswall thickness as well as the increase in body weight and the level of cholesterol and triglyceride. The extracts also hadno effect on the concentrations of estrogen and growth hormone in the serum. However, the increase in alkalinephosphatase activity, which leads to protection against the bone loss caused by ovariectomy, was noted on administrationof the extract. Therefore, it seemed that the extracts of C. wilfordii and P. umbrosa Turcz. had no estrogenic effect in rats.Keywords: Cynanchum wilfordii, Phlomis umbrosa Turcz, isoflavone, estrogen, growth hormone ovariectomized rat

The present study was aimed to investigate the effects of MB12662, a synthetic dunnione compound, on cisplatin-induced vomiting reflexes and intestinal, renal, immune system, and hematopoietic toxicities in ferrets and mice, respectively. Male ICR mice were orally administered MB12662 (5, 10, 25 or 50 mg/kg) for 10 days, during which intraperitoneally challenged with cisplatin (3.5 mg/kg) from day 4 to 7, and sacrificed on day 10 for the pathological examination. Male ferrets were orally administered MB12662 (25, 50 or 100 mg/kg) for 7 days, subcutaneously challenged with cisplatin (5 mg/kg), and monitored for vomiting reflexes and survival of the animals. Four-day injection of cisplatin (3.5 mg/kg) to mice caused body weight loss and degeneration and atrophy of intestinal villi, reducing villi/crypt ratio to a half level of control animals. Cisplatin also induced renal and hepatic toxicities, and depletion of splenocytes and bone marrow progenitor cells. The systemic toxicities including decreased villi/crypt ratio, immune system atrophy, splenocyte depletion, and decreased cellularity in bone marrow were improved by MB12662. Cisplatin (5 mg/kg) induced retching and emetic responses of ferrets, which were remarkably attenuated by MB12662 in a dose-dependent manner. All the ferrets pretreated with MB12662 survived the challenge of cisplatin, in comparison with 40% mortality in vehicle-treated animals, and blood parameters of nephrotoxicity and hepatotoxicity were markedly recovered. It is expected that MB12662 could be a candidate for the body protection against burden, including emesis, of chemotherapeutic agents.

High pressure (HP) is recognized as the alternative process technology of heat treatment in food production. Although milk was the first food product to be treated with HP, the allergenicity of HP-treated milk remains to be elucidated. Thus, this study was conducted to investigate the effects of HP treatment on the alleviation of atopic dermatitis (AD) caused by milk in a mouse model. To investigate the effect of HP treatment on the alleviation of AD, Balb/c mice were fed HP-treated milk orally for 7 days following induction of AD with 1-chloro-2,4-dinitro-benzene (DNCB). Mice fed with HP-treated milk exhibited markedly lower serum IgE level than DNCB-treated mice. These results suggest that HP may be applicable as an alternative process for the development of hypoallergenic milk products.

Lactococcus lactis subsp. lactis WK11 was introduced as a starter in kimchi fermentation to investigate the role of starter cultures in extending the shelf life of kimchi. Kimchi, with an initial inoculum size of 107 CFU/g as well as control kimchi without the starter culture, were prepared and fermented for 56 days at 8°C. The addition of L. lactis prolonged the optimal fermentation period (pH 4.2, acidity 0.6–0.8%) up to more than 2 times. The starter culture, L. lactis WK11, persisted throughout the kimchi fermentation, as confirmed by regular monitoring of the L. lactis population by PCR-DGGE. Taken together, our results suggest that L. lactis starter culture may be useful for extending the shelf-life of kimchi without adverse effects of kimchi with a longer shelf life.

Allium hookeri, a member of the onion family, has long been mainly cultivated for food and medicinal use in Southeast Asian countries, owing to its various biological properties. However, no studies of the anti-oxidative effects of fermented A. hookeri root extracts have been conducted to date. Therefore, this study investigated the effect of different starter cultures on the antioxidant activities of hot water extract of A. hookeri root by using the following five strains: Lactobacillus acidophilus, Bifidobacterum longum, Enterococcus faecium, Streptococcus thermophilus, and Saccharomyces cerevisiae. DPPH and ABTS radical scavenging activity, total phenolic acid contents, and total antioxidant capacity were higher in the hot water extract of A. hookeri root fermented with starter cultures than those of A. hookeri root. Among hot water extract of A. hookeri root fermented with starter cultures, fementing with S. cerevisiae showed the highest antioxidant activities. The results of this study provide new evidence of the anti-oxidative properties of A. hookeri root with starter cultures, indicating that it may be highly valuable as a natural product owing to its high-quality functional components.

Anti-adipogenic and anti-obesity effects of a green coffee been extract (GCE) from unroasted Coffea canephora Robusta beans were investigated in vitro and in vivo. Insulin-induced adipogenesis in 3T3-L1 preadipocytes was inhibited by 25.6 and 87.9% following treatment with 100 and 500 μg/mL GCE, respectively. Expression of adipogenesis-specific genes was down-regulated. Body weight gains of C57BL/6 mice fed a high-fat diet (HFD) were suppressed in a concentration-dependent manner using GCE (0.3 or 1% in the diet), exhibiting a marked lower gain in 1% GCE-fed mice than in normal diet-fed animals. Decreases in body fat weights and the size of adipocytes were confirmed. GCE supplementation reduced concentrations of blood lipids and leptin, and attenuated hepatic steatosis. GCE inhibits adipogenesis by modulating cell signaling and improves dietary obesity by reducing lipid accumulation and the size of adipocytes.

Plasma, formed by ionization of gas molecules or atoms, is the most abundant form of matter and consists of highly reactive physicochemical species. In the physics and chemistry fields, plasma has been extensively studied; however, the exact action mechanisms of plasma on biological systems, including cells and humans, are not well known. Recent evidence suggests that cold atmospheric plasma (CAP), which refers to plasma used in the biomedical field , may regulate diverse cellular processes, including neural differentiation. However, the mechanism by which these physicochemical signals, elicited by reactive oxygen and nitrogen species (RONS), are transmitted to biological system remains elusive. In this study, we elucidated the physicochemical and biological (PCB) connection between the CAP cascade and Trk/Ras/ERK signaling pathway , which resulted in neural differentiation. Excited atomic oxygen in the plasma phase led to the formation of RONS in the PCB network, which then interacted with reactive atoms in the extracellular liquid phase to form nitric oxide (NO). Production of large amounts of superoxide radical ( O2−) in the mitochondria of cells exposed to CAP demonstrated that extracellular NO induced the reversible inhibition of mitochondrial complex IV. We also demonstrated that cytosolic hydrogen peroxide, formed by O2− dismutation, act as an intracellular messenger to specifically activate the Trk/Ras/ERK signaling pathway. This study is the first to elucidate the mechanism linking physicochemical signals from the CAP cascade to the intracellular neural differentiation signaling pathway, providing physical, chemical and biological insights into the development of therapeutic techniques to treat neurological diseases.

Herbicidin A is a potent herbicide against dicotyledonous plants as well as an antibiotic against phytopathogens. In this study, fermentation parameters for herbicidin A production in submerged culture of

Background Allium hookeri (AH) is widely consumed as a vegetable and herbal medicine in southeastern Asia. AH has been reported antioxidant, antimicrobial, improvement of bone health and antidiabetic effects. In the present study, we investigated the inhibitory effect of a methanol extract of AH root (AHE) on inflammatory response in lipopolysaccharide (LPS)-induced RAW264.7 cells. Methods Initially, characterization of organic sulfur compounds in AHE was determined using high performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS). Cells were incubated with LPS and AHE for 24 h. The productions of nitric oxide (NO), reactive oxygen species (ROS), and inflammation-related cytokines were examined. Gene and protein expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were assessed by polymerase chain reaction and Western blotting. Key factor, nuclear factor kappa B (NF-κB) was also determined. Results AHE contained organosulfur compounds such as alliin and S-allylcysteine by HPLC-ESI-MS. AHE significantly inhibited NO, ROS, and cytokines production in LPS-induced RAW264.7 cells. In addition, AHE treatment inhibited iNOS and COX-2 mRNA and protein levels, leading to a decrease in iNOS-derived NO level. Furthermore, NF-κB activation was, at least in part, suppressed by AHE treatment. Conclusion Our data suggest that AHE treatment inhibits the inflammation condition through suppression of iNOS and COX-2 expression via NF-κB down-regulation.

The occurrence of atopic dermatitis (AD), a chronic inflammatory skin disease, has been increasing steadily in children and adults in recent decades. In this study, we evaluated the ability of the lactic acid bacterium Weissella cibaria WIKIM28 isolated from gatkimchi, a Korean fermented vegetable preparation made from mustard leaves, to suppress the development of AD induced by 2,4-dinitrochlorobenzene in a murine model. Oral administration of W. cibaria WIKIM28 reduced AD-like skin lesions, epidermal thickening, and serum immunoglobulin E levels. Furthermore, the production of type 2 helper T (Th2) cytokines such as interleukin (IL)-4, IL-5, and IL-13 decreased in peripheral lymph node cells. Moreover, the intake of W. cibaria WIKIM28 increased the proportion of CD4+CD25+Foxp3+ regulatory T (Treg) cells in mesenteric lymph nodes (MLNs) and IL-10 levels in polyclonally stimulated MLN cells. In conclusion, the oral administration of W. cibaria WIKIM28 isolated from gatkimchi ameliorated AD-like symptoms by suppressing allergic Th2 responses and inducing Treg responses. These results suggest that W. cibaria WIKIM28 may be applicable as a probiotic for the prevention and amelioration of AD.

This study examined the potential immunomodulatory effects of Kimchi, a traditional fermented Korean vegetable, in healthy Chinese college students. The four-week clinical-trial (randomized, open-label, prospective, controlled) was followed by a one week wash-out period. Healthy Chinese college students (over 20 years of age with a body mass index of 18.5-23.0 kg/m 2 ) volunteered for this study. Forty-three students were randomly classified into two groups, Kimchi (n = 21, supplemented with 100 g of Kimchi per day) or non-Kimchi (n = 22, supplemented with 100 g of radish per day, control) groups. During the four-week intervention period, students were asked to maintain their usual diet and activity, and instructed not to take any medications, functional food products, or dietary supplements. Anthropometrics, nutritional intake, and blood immune parameters (lymphocyte subsets, cytokines, and immunoglobulins) were measured before and after the four weeks of intervention. Thirty-nine students (19 in the Kimchi group, 20 in the non-Kimchi group) finished the study. After the intervention, no significant changes were observed in lymphocyte subsets (T-cell, B-cell, NK cell), pro-inflammatory cytokines (IL-6, TNF- α), anti-inflammatory cy tokines (IL-4 and IL-10), and immunoglobulins (Ig A, G, and M) between groups in either the Kimchi or non-Kimchi. These results suggest that the short-term consumption of Kimchi has no immunomodulatory effects in healthy Chinese college students.

This study was conducted to estimate the contents of heavy metals in salted vegetable foods fromdiverse origin in Korea which were sold in Korea. The levels of heavy metals were determined using an ICP-MS. Thevalues of metals [minimum~maximum (mean), mg/kg] in kimchi were as follows; Pb 0.0~0.074(0.018), Cd 0.0~0.027(0.004), As 0.0~0.024(0.002), Hg 0~0.002(0.0). The weekly average intakes of lead, cadmium, arsenic and mercuryfrom kimchi take 0.06~0.13% PTWI (Provisional Tolerable Weekly Intakes) that the FAO/WHO Joint Food Additiveand Contaminants Committee has set to evaluate their safeties. Key words : Safety, Fermented vegetable foods, lead, cadmium 중금속은 비중이 4 이상 되는 금속원소로서 86종 가량이 있으며, 이들 중 약 27종만이 생체의 기능유지에 필수적이며, 필요량에서 부족하거나 다량일 경우 건강상의 장해를 일으킨다. 대다수의 금속은 영양학적으로 무의미하며 그 중에서 동·식물체에 유해성이 큰 중금속으로는 수은(Hg), 카드뮴(Cd), 납(Pb), 비소(As), 주석(Sn), 아연(Zn),구리(Cu), 크롬(Cr), 망간(Mn) 등이 이에 해당한다. 이들금속의 농도, 존재하는 화학성, 배설 속도, 건강 상태 등다양한 인자에 따라 생체에서의 유해 여부에 차이가 있다. 그런데 중금속은 자신이 가지고 있는 독성뿐만 아니라 축적성이 있어 먹이 연쇄에 따라 크게 농축된다. 특히, 수은, 카드뮴, 납은 식품 중에서 공통적으로 볼 수 있는 독성 물질로 생체 조직과 강한 결합을 하여 생체 내에서 축적되어 천천히 제거되는 유해 금속이다

Since oils and fats can induce metabolic syndrome, leading to cardiovascular and cerebrovascular diseases, the present study was performed to find out whether the plant oils affect the cerebral hemorrhage in stroke-prone spontaneously hypertensive (SHR-SP) rats.From 47 days of age, male SHR-SP rats were given drinking water containing 1% NaCl to induce hypertension, and simultaneously fed semi-purified diets containing 10% perilla oil, canola oil, or shortening. The onset time of convulsion following cerebral hemorrhage was recorded, and the areas of hemorrhage and infarction were analyzed in the stroke brains.In comparison with 58-day survival of SHR-SP rats during feeding NaCl alone, perilla oil extended the survival time to 68.5 days, whereas canola oil shortened it to 45.7 days. Feeding perilla oil greatly reduced the total volume of cerebral hemorrhage from 17.27% in the control group to 4.53%, while shortening increased the lesions to 21.23%. In a microscopic analysis, perilla oil also markedly decreased the hemorrhagic and infarction lesions to 1/10 of those in control rats, in contrast to an exacerbating effect of shortening. In blood analyses, perilla oil reduced blood total cholesterol and low-density lipoproteins which were increased in SHR-SP, but canola oil further increased them and markedly lowered platelet counts.Perilla oil delayed and attenuated cerebral hemorrhage by improving hyperlipidemia in hypertensive stroke animals, in contrast to the aggravating potential of canola oil and shortening. It is suggested that perilla oil should be the first choice oil for improving metabolic syndrome in hypertensive persons at risk of hemorrhagic stroke.

Allium hookeri (AH) is widely consumed as a herbal medicine. It possesses biological activity against metabolic diseases. The objective of this study was to investigate effects of AH root water extract (AHR) on adipogenesis in 3T3-L1 cells and in high-fat diet (HFD)-induced obese mice. AHR inhibited lipid accumulation during adipocyte differentiation by downregulation of gene expression, such as hormone sensitive lipase (HSL), lipoprotein lipase (LPL) and an adipogenic gene, CCAAT/enhancer binding protein-&alpha, in 3T3-L1 preadipocytes. Oral administration of AHR significantly suppressed body weight gain, adipose tissue weight, serum leptin levels, and adipocyte cell size in HFD-induced obese mice. Moreover, AHR significantly decreased hepatic mRNA expression levels of cholesterol synthesis genes, such as 3-hydroxy-3-methylglutaryl CoA reductase, sterol regulatory element-binding transcription factor (SREBP)-2, and low-density lipoprotein receptor, as well as fatty acid synthesis genes, such as SREBP-1c and fatty acid synthase. Serum triglyceride levels were also lowered by AHR, likely as a result of the upregulating gene involved in fatty acid &beta, oxidation, carnitine palmitoyltransferase 1a, in the liver. AHR treatment activated gene expression of peroxisome proliferator-activated receptor-&gamma, which might have promoted HSL and LPL-medicated lipolysis, thereby reducing white adipose tissue weight. In conclusion, AHR treatment can improve metabolic alterations induced by HFD in mice by modifying expression levels of genes involved in adipogenesis, lipogenesis, and lipolysis in the white adipose tissue and liver.

This study was conducted to investigate the quality characteristics of kimchi with Allium hookeri powder (AHP) added during eight weeks fermentation at 4°C. AHP was added to salted cabbage at concentrations of 0, 0.1, 1, and 5% (w/w). The quality characteristics of the AHR-added kimchi were determined by measuring pH, acidity, salinity, reducing sugar, microbial amounts, and sensory properties. The vitamin B1 and vitamin C contents of the Allium hookeri root were 0.04 mg/100 g and 5.76 mg/100 g, respectively. As for the mineral contents, the K content was highest, followed by the Ca, Mg, and Fe contents. The pH was higher in the kimchi with 0.1%, 1%, and 5% AHP than in the kimchi without AHP during the eight-week fermentation. The salinity ranged from 2.02% to 2.37% over the eight weeks. The microbial cells and lactobacilli increased rapidly throughout the fermentation in the exponential phase and hardly increased in the stationary phase. In the sensory evaluations, the overall acceptance, taste, and texture of the kimchi with 1% AHP added were highest. In conclusion, the kimchi with 0.1%, 1%, and 5% AHP generally showed better quality than the kimchi without AHP. Especially, the kimchi with 1% AHP had the best scores in quality and overall acceptance during the fermentation.

Scope The gut microbiota has been linked to diet-induced obesity, and microorganisms that influence obesity have important health implications. In this study, the anti-obesity effects of two Lactobacillus plantarum strains (DSR M2 and DSR 920) isolated from kimchi are investigated. Methods and results Mice are fed a normal or high-fat diet with or without DSR M2 and DSR 920 (DSR, 1 × 109 CFU d-1 ) for 12 weeks. DSR improves the obesity state, as evidenced by the i) suppressed obesity-related markers, e.g., gains in body weight and fat mass, ii) reduced serum and liver triglyceride levels, iii) upregulated β-oxidation and downregulated lipogenesis-related genes in the liver, iv) reduced serum leptin levels, v) altered microbial communities, vi) increased regulatory T cell immunity, and vii) suppressed inflammatory response. In addition, correlation analysis shows that Akkermansia muciniphila and the genus Anaerostipes, which are increased in the DSR group, are negatively correlated with obesity-related markers, but Mucispirillum schaedleri, which is increased in the high-fat-diet (HFD) group, is positively correlated with serum leptin level. Conclusion Lactobacillus plantarum DSR M2 and DSR 920 are candidate probiotics for the prevention and amelioration of obesity.

Licorice (Glycyrrhiza glabra Linne) is a well-known medicinal plant and glabridin is an isoflavan isolated from licorice. In this study, we investigated the anti-obesity effect of glabridin and glabridin-rich supercritical fluid extract of licorice (LSC). Glabridin effectively inhibited adipogenesis in 3T3-L1 cells. Moreover, LSC showed inhibitory effect on adipogenesis in a dose-dependent manner. The inhibitory effect of LSC resulted from inhibiting the induction of the transcriptional factors CCAAT enhancer binding protein alpha and peroxisome proliferator-activated receptor gamma. Then we fed mice with high-fat diet containing none, 0.1% and 0.25% LSC for 8weeks to explore the anti-obesity effect of LSC in vivo. LSC significantly reduced weight gain by high-fat diet in a dose-dependent manner. The reductions of the hypertrophy of white adipose tissue and of fat cell size were also observed. In the liver, LSC supplementation effectively inhibited high-fat diet-induced hepatic steatosis through downregulation of gluconeogenesis related phosphoenolpyruvate carboxykinase and glucose 6-phosphatase and upregulation of the β-oxidation related carnitine palmitoyltransferase 1. Taken together, our results suggest that glabridin and glabridin-rich licorice extract would be effective anti-obesity agents.

A novel halophilic bacterium, strain K7(T), was isolated from kimchi, a traditional Korean fermented food. The strain is Gram-positive, motile, and produces terminal endospores. The isolate is facultative aerobic and grows at salinities of 0.0-25.0% (w/v) NaCl (optimum 10-15% NaCl), pH 5.5-8.5 (optimum pH 7.0-7.5), and 15-42°C (optimum 37°C). The predominant isoprenoid quinone in the strain is menaquinone-7 and the peptidoglycan of the strain is meso-diaminopimelic acid. The major fatty acids of the strain are anteisio-C15:0, iso-C15:0, and, C16:0 (other components were10.0%), while the major polar lipids are diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, and three unidentified lipids. A phylogenetic analysis of 16S rRNA gene sequence similarity showed that the isolated strain was a cluster of the genus Gracilibacillus. High levels of gene sequence similarity were observed between strain K7(T) and Gracilibacillus orientalis XH-63(T) (96.5%), and between the present strain and Gracilibacillus xinjiangensis (96.5%). The DNA G+C content of this strain is 37.7 mol%. Based on these findings, strain K7(T) is proposed as a novel species: Gracilibacillus kimchii sp. nov. The type strain is K7(T) (KACC 18669(T); JCM 31344(T)).

The oxidation of low-density lipoproteins (oxLDL) is considered as a key step in the development of atherosclerosis. Oxysterols such as 7-ketocholesterol (7-KC) have been reported to be partially responsible for the cytotoxicity of oxLDL and induce endoplasmic reticulum (ER) stress which eventually causes apoptosis. We aimed to investigate whether ER stress is induced by 7-KC and can be prevented by the kimchi methanol extract (KME) in RAW264.7 macrophage cells. ER stress induced by 7-KC was characterized by the activation of ER stress markers (GRP78, CHOP, and ATF6). ER stress could aggravate macrophage apoptosis, as assessed by caspase-3 activity. ER stress and apoptosis were inhibited by the KME and glutathione (GSH). We conclude that 7-KC causes aberrant ER stress and apoptosis, all of which are inhibited by KME and GSH. The inhibitory expression of ER stress markers highlights its new protective role against oxLDL-induced ER stress, apoptosis, and subsequent atherosclerosis.

A Gram-positive, aerobic, non-motile and extremely halophilic bacterial strain, designated K9(T), was isolated from kimchi, a Korean fermented food. The strain was observed as endospore-forming rod-shaped cells showing oxidase and catalase activity. It was found to grow at 10.0-30.0 % (w/v) NaCl (optimum, 15.0-20.0 %), pH 7.0-8.0 (optimum, pH 7.5) and 15-40 °C (optimum, 30 °C). The polar lipids of strain K9(T) were identified as phosphatidylglycerol, three unidentified phospholipids and an unidentified glycolipid. The isoprenoid quinone was identified as menaquinone-7. The major cellular fatty acids (>20 % of the total) were found to be anteisio-C15:0 and anteisio-C17:0. The cell wall peptidoglycan composition was determined to contain meso-diaminopimelic acid. The G + C content of genomic DNA was determined to be 48.2 mol %. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that the isolated strain is closely related to Lentibacillus salinarum AHS-1(T) (96.7 % sequence similarity). Based on its phenotypic, chemotaxonomic and phylogenetic data, strain K9(T) is considered to represent a novel species of the genus Lentibacillus, for which the name Lentibacillus kimchii sp. nov., is proposed. The type strain is K9(T) (=KACC 18490(T) = JCM 30234(T)).