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Dextran, a homologous polysaccharide with the main chain of glucose units composed by an α-1,6 glycosidic bond, is synthesized from dextrin by dextran-dextrinase (DDase), a transglucosidase, derived from acetic acid bacteria Gluconobacter oxydans and lactic acid bacteria (LAB). The effective culture conditions were examined for producing dextran via bio-conversion with acetic acid bacteria (AAB) demonstrating DDase activity in various medium compositions during 0 to 7 days with or without glycerol addition (2%, v/v) and different degrees of dextrin polymerization (D.E.) based on the addition level (1,5%, w/v). On day 7, the G. oxydans growth was almost tripled in presence of glycerol as observed via a cell growth curve (OD). After culturing for 7 days, the pH decreased from 6 to 3.1-3.5, and the acidity increased from 0.12% to 0.4-0.62%, depending on the dextrin D.E. and the addition level. The reducing sugar decreased continuously. The medium containing 5% dextrin showed shear-thinning characteristics. The apparent viscosity of the 5% dextrin DE4-7 culture solution was 5.6 mPa·s, which was similar to that of the 20% dextran aqueous solution. The analysis of constituent saccharides contained in the culture medium (HPAEC-PAD) showed a substance with a high degree of polymerization. 1H-NMR analysis showed that α-1,6 glycosidic bond existed as the intermolecular bond of this substance. Therefore, efficient production of dextran was possible by culturing in a medium containing 5% dextrin and glycerol during culture of AAB.

A γ-aminobutyric acid (GABA)-producing microorganism was isolated from galchi (hairtail fish, Trichiurus lepturus) jeotgal, a Korean salted and fermented seafood. The G144 isolate produced GABA excessively when incubated in MRS broth containing monosodium glutamate (MSG, 3%, w/v). G144 was identified as Lactobacillus brevis through 16S rRNA and recA gene sequencing. gadB and gadC encoding glutamate decarboxylase (GAD) and glutamate/GABA antiporter, respectively, were cloned and gadB was located downstream of gadC. The operon structure of gadCB was confirmed by reverse transcription (RT)-polymerase chain reaction. gadB was overexpressed in Escherichia coli and recombinant GAD was purified and its size was 54.4 kDa as evidenced by SDS-PAGE results. Maximum GAD activity was observed at pH 5.0 and 40℃ and the activity was dependent on pyridoxal 5''-phophate. The Km and Vmax of GAD were 8.6 mM and 0.01 mM/min, respectively.

Lactobacillus zymae GU240 was previously isolated from Kimchi, a Korean fermented vegetable, as a strong GABA producer. The strain showed β-galactosidase (β-Gal) activity on MRS agar plates with X-gal. When growth and β-Gal activities of GU240 were measured using MRS (glucose, 2%, w/v) and MRSL (lactose, 2%, w/v) broths, cells were found to grow slowly in MRSL, and the β-Gal activity (36 units at 4 h) was lower than that of cells grown in MRS (94 units at 16 h). The highest OD600 value of the culture in MRS was 1.6 at 24 h at 37℃, whereas that of the culture in MRSL was 0.6 at 16 h. β-Gal activity of the culture in MRS reached the maximum (95.6 u/ml) at 16 h, decreased thereafter, and was not detected at 48 h. β-Gal activity for culture in MRSL reached its highest (36 u/ml) at 4 h and decreased gradually, but some activity (11.05 u/ ml) still remained at 72 h. The structural gene encoding β-Gal in L. zymae GU240 was cloned as a 3.1 kb fragment, and DNA sequencing confirmed the presence of complete lacLM genes. lacLM genes from L. zymae GU240 showed 98-99% homologies in nucleotide sequences with other lacLM genes from L. brevis. Reverse transcription (RT)-PCR confirmed the operon structure of lacLM. The results indicated that L. zymae GU240 might be in the process of losing the ability to grow rapidly on lactose-containing medium, such as milk, due to adaptations to plant environments, including kimchi.

Perilla seed powder (PSP) was stored at 25 °C, 35 °C, and 45 °C for 8 weeks. Changes in the metabolite profiles of the powders, including fatty acids, were monitored. Correlations between these changes and quality parameters, including lipid oxidation, color, and antioxidant activity, were analyzed to evaluate the effects of storage duration and temperature on PSP quality. Acid values increased significantly with the duration of storage, but not with temperature. Multivariate statistical analysis was performed to identify differences among the metabolite profiles. The PSP sample stored for 1 week at 45 °C and all samples stored at 25 °C and 35 °C were grouped separately from the control and samples stored at 45 °C for more than 4 weeks. Among the many metabolites associated with these differences, lysophosphatidylethanolamines, tocopherol, sitosterol, tryptophan, 12-hydroxyjasmonic acid glucoside, and maltose correlated negatively with quality parameters with the exception of L* and antioxidant activity. Luteolin, apigenin, luteolin 4′-methyl ester, citric acid, isocitric acid, 9(S)-HPODE, and 3,5-octadien-2-one correlated positively with quality. Although the quantities of some antioxidants and lipids decreased during storage, the results suggested that the quality of PSP samples stored at 25 °C, 35 °C, and 45 °C for 8 weeks was acceptable. This was because lipid oxidation promoted by the storage environment was limited by antioxidants in the samples. These metabolites could be useful for monitoring changes in PSP quality. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s13197-019-04056-2) contains supplementary material, which is available to authorized users.

Pasteurization‐mediated delayed kimchi ripening and regression analysis for shelf life estimation were investigated. Various initial kimchi microbial communities were simplified to lactic acid bacteria Leuconostoc sp. and Lactobacillus sp. over time, with concomitant pH decrease from 6.39 to 4.34 and acidity increase from 0.06% to 0.35%. Other quality characteristics (organic acid, carbon dioxide, and microbial population) also changed, exhibiting high intercorrelation. Pasteurization decreased the initial bacterial counts from 5.20 to 1.92 log CFU/g, thereby delaying the change in quality characteristics (pH, acidity, organic acid, microbial population, carbon dioxide, and microbial community); however, the texture did not differ significantly (p

Jang S, Jang S, Kim SY, et al. J Inflamm Res. 2021;14:4299–4312. The authors have advised the author list on page 4299 is incorrect. The correct author list and affiliations are as follows. Soyeon Jang1,* Soyoung Jang1,* Si-Yong Kim1 Jiwon Ko1 Eungyung Kim2 Ji Yeong Park1 Hyejin Hyung1 Jin Hong Lee1 Su-Geun Lim3 Sijun Park3 Junkoo Yi4 Heon-Jin Lee5 Myoung Ok Kim2 Hyun-Shik Lee1 Zae Young Ryoo1 1School of Life Sciences, BK21 FOUR KNU Creative BioResearch Group, Kyungpook National University, Daegu, Korea; 2Department of Animal Science and Biotechnology, Kyungpook National University, Daegu, Korea; 3School of Life Sciences, Kyungpook National University, Daegu, Korea; 4Gyeongsangbukdo Livestock Research Institute, Yeongju, Korea; 5Department of Microbiology and Immunology, School of Dentistry, Kyungpook National University, Daegu, 41940, Korea *These authors contributed equally to this work Read the original article

Isomaltooligosaccharide (IMO), considered to be a prebiotic, reportedly has health effects, particularly in terms of digestion; however, the prebiotic effects of IMOs depend largely on the degree of polymerization. Currently, IMOs are commercially produced using transglucosidase (TG) derived from Aspergillus niger. Here, we report a novel Thermoanaerobacter thermocopriae-derived TG (TtTG) that can produce long-chain IMOs (L-IMOs) using maltodextrin as the main substrate. A putative carbohydrate-binding gene comprising carbohydrate-binding module 35 and glycoside hydrolase family 15 domain was cloned and successfully overexpressed in Escherichia coli BL21 (DE3) cells. The resulting purified recombinant enzyme (TtTG) had a molecular mass of 94 kDa. TtTG displayed an optimal pH of 4.0 (higher than that of commercial TG) and an optimal temperature of 60 °C (same as that of commercial TG). TtTG also enabled the synthesis of oligosaccharides using various saccharides, such as palatinose, kojibiose, sophorose, maltose, cellobiose, isomaltose, gentiobiose, and trehalose, which acted as specific acceptors. TtTG could also produce a medium-sized L-IMO, different from that by dextran-dextrinase and TG, from maltodextrin, as the sole substrate. Thus, the novel combination of maltodextrin and TtTG shows potential as an effective method for commercially producing L-IMOs with improved prebiotic effects.

The effects of five different drying conditions on kimchi powder quality were determined by comparatively analyzing their physicochemical characteristics, volatile profile, and sensory evaluations. The moisture content of the kimchi powder obtained by each method was

The efficiency factor (η) and the instability factor ([Formula: see text] in deformation processing maps are regarded as the reliable indices of formability during high-temperature deformation. Deformation processing maps are primarily based on strain rate sensitivity ( m) and are usually created by high-temperature compression tests. To analyze the effect of the mode of flow on the deformation processing map, deformation processing maps based on both compressive and tensile flow curves for 304L stainless steel were determined and compared in the current study. As the instantaneous strain rate varies during both the tensile and compression tests when a constant crosshead speed is used, strain rate compensated deformation processing maps have been determined and compared. In addition, the frictional effect of barreling during compression testing on the deformation processing map has been analyzed. Both deformation processing maps based on either compressive or tensile flow curves are estimated to be complementary.

Flexible roll forming is an advanced sheet metal forming process that allows for the production of variable cross-section profiles. Longitudinal bow is one of the major shape defects found in roll-formed products. To characterize the degree of longitudinal bow during flexible roll forming, experiments were conducted on three different blank shapes: trapezoid, convex and concave. Symmetric U-sections with variable cross-sections were roll formed using the three sheet materials with different strengths from each blank shape. The effects of process variables on the longitudinal strain and longitudinal bow were analyzed both experimentally and by using finite element simulations based on ABAQUS-Implicit 6.14. The results show that the transversal nonuniformity of the longitudinal strain is one of the fundamental causes of longitudinal bow in roll-formed products. The bow height as a function of blank shape increases in the order of concave, trapezoid, and convex. The bow characteristics on the basis of blank shapes and material parameters provides a better understanding of the non-uniform longitudinal strain and bow height during flexible roll forming with high reliability. Furthermore, to reduce the longitudinal bowing, leveling roll is introduced. The effect of leveling roll on longitudinal bowing was investigated experimentally. The results show that longitudinal bowing was reduced with the use of three blank shapes when leveling roll was applied to a flexible roll forming process.

In the Chromosome-Centric Human Proteome Project (C-HPP), false-positive identification by peptide spectrum matches (PSMs) after database searches is a major issue for proteogenomic studies using liquid-chromatography and mass-spectrometry-based large proteomic profiling. Here we developed a simple strategy for protein identification, with a controlled false discovery rate (FDR) at the protein level, using an integrated proteomic pipeline (IPP) that consists of four engrailed steps as follows. First, using three different search engines, SEQUEST, MASCOT, and MS-GF+, individual proteomic searches were performed against the neXtProt database. Second, the search results from the PSMs were combined using statistical evaluation tools including DTASelect and Percolator. Third, the peptide search scores were converted into E-scores normalized using an in-house program. Last, ProteinInferencer was used to filter the proteins containing two or more peptides with a controlled FDR of 1.0% at the protein level. Finally, we compared the performance of the IPP to a conventional proteomic pipeline (CPP) for protein identification using a controlled FDR of1% at the protein level. Using the IPP, a total of 5756 proteins (vs 4453 using the CPP) including 477 alternative splicing variants (vs 182 using the CPP) were identified from human hippocampal tissue. In addition, a total of 10 missing proteins (vs 7 using the CPP) were identified with two or more unique peptides, and their tryptic peptides were validated using MS/MS spectral pattern from a repository database or their corresponding synthetic peptides. This study shows that the IPP effectively improved the identification of proteins, including alternative splicing variants and missing proteins, in human hippocampal tissues for the C-HPP. All RAW files used in this study were deposited in ProteomeXchange (PXD000395).

본 연구는 대학생의 데이트폭력 실태를 살펴보고, 데이트폭력에 영향하는 요인을 파악하여 데이트 폭력의 효율적인 예방과 간호 중재 개발에 기초 자료를 제공하고자 실시되었다. 연구는 D시 소재 6 개 대학(4년제 대학교 4곳, 2~3년제 전문대학 2곳)에 재학 중인 남, 녀 대학생 209명을 대상으로 온/ 오프라인을 통해 설문지 조사를 실시하였다. 데이트폭력에 영향하는 요인을 알아보기 위하여 아동기 학대경험, 성인애착, 관계통제를 측정하였다. 자료는 SPSS/WIN 21.0 통계프로그램을 이용하여 기술통 계, Mann-Whitney U test, Kruskal-Wallis test, Spearman's rho, 붓스트랩 회귀분석 방식을 사용하여 분석하였다. 연구결과 데이트폭력에 영향하는 요인으로 아동기 학대경험과 관계통제를 확인하였다. 따라서 데이트폭력 위험군의 조기 발견을 위한 도구 개발과 올바른 이성에 대한 태도, 갈등 상황에서 의 비폭력적인 해결 방법 등이 포함된 대학생의 건전한 이성교제를 위한 간호중재 프로그램이 개발 되어야 할 필요가 있다.

Fibrinolytic enzyme genes (aprE2, aprE176, and aprE179) were introduced into the Bacillus subtilis 168 chromosome without any antibiotic resistance gene. An integration vector, pDG1662, was used to deliver the genes into the amyE site of B. subtilis 168. Integrants, SJ3-5nc, SJ176nc, and SJ179nc, were obtained after two successive homologous recombinations. The integration of each fibrinolytic gene into the middle of the amyE site was confirmed by phenotypes (Amy(-), Spec(S)) and colony PCR results for these strains. The fibrinolytic activities of the integrants were higher than that of B. subtilis 168 by at least 3.2-fold when grown in LB broth. Cheonggukjang was prepared by inoculating each of B. subtilis 168, SJ3-5nc, SJ176nc, and SJ179nc, and the fibrinolytic activity of cheonggukjang was 4.6 ± 0.7, 10.8 ± 0.9, 7.0 ± 0.6, and 8.0 ± 0.2 (U/g of cheonggukjang), respectively at 72 h. These results showed that construction of B. subtilis strains with enhanced fibrinolytic activities is possible by integration of a strong fibrinolytic gene via a marker-free manner.

A gamma-aminobutyric acid (GABA)-producing microorganism was isolated from jeot-gal (anchovy), a Korean fermented seafood. The isolate, A156, produced GABA profusely when incubated in MRS broth with monosodium glutamate (3% (w/v)) at 37°C for 48 h. A156 was identified as Lactobacillus sakei by 16S rRNA gene sequencing. The GABA conversion yield was 86% as determined by GABase enzyme assay. The gadB gene encoding glutamate decarboxylase (GAD) was cloned by PCR. gadC encoding a glutamate/GABA antiporter was located immediately upstream of gadB. The operon structure of gadCB was confirmed by RT-PCR. gadB was overexpressed in Escherichia coli BL21(DE3) and recombinant GAD was purified. The purified GAD was 54.4 kDa in size by SDS-PAGE. Maximum GAD activity was observed at pH 5.0 and 55°C and the activity was dependent on pyridoxal 5'-phosphate. The Km and Vmax of GAD were 0.045 mM and 0.011 mM/min, respectively, when glutamate was used as the substrate.

A cryptic plasmid, pJY33, from Weissella cibaria 33 was characterized. pJY33 was 2365 bp in size with a GC content of 41.27% and contained two putative open reading frames (ORFs). orf1 encoded a putative hypothetical protein of 134 amino acids. orf2 was 849 bp in size, and its putative translation product exhibited 87% identity with a replication initiation factor from a plasmid from W. cibaria KLC140. A Weissella - Escherichia coli shuttle vector, pJY33E (6.5 kb, Em r ), was constructed by ligation of pJY33 with pBluescript II SK(−) and an erythromycin resistance gene (Em r ). pJY33E replicated in Lactococcus lactis , Leuconostoc citreum , Lactobacillus brevis, Lactobacillus plantarum, and Weissella confusa . A single-stranded DNA intermediate was detected from Lb. brevis 2.14 harbouring pJY33E, providing evidence for rolling-circle replication of pJY33. Most Lb. brevis 2.14 cells (85.9%) retained pJY33E after one week of daily culturing in MRS broth without Em. An aga gene encoding α-galactosidase (α-Gal) from Leuconostoc mesenteroides was successfully expressed in Lb. brevis 2.14 using pJY33E, and the highest level of α-Gal activity (36.13 U/mg protein) was observed when cells were grown on melibiose.

Human Proteome Project aims to map all human proteins including missing proteins as well as proteoforms with post translational modifications, alternative splicing variants (ASVs), and single amino acid variants (SAAVs). neXtProt and Ensemble databases are usually used to provide curated information on human coding genes. However, to find these proteoforms, we (Chr #11 team) first introduce a streamlined pipeline using customized and concatenated neXtProt and GENCODE originated from Ensemble, with controlled false discovery rate (FDR). Because of large sized databases used in this pipeline, we found more stringent FDR filtering (0.1% at the peptide level and 1% at the protein level) to claim novel findings, such as GENCODE ASVs and missing proteins, from human hippocampus data set (MSV000081385) and ProteomeXchange (PXD007166). Using our next generation proteomic pipeline (nextPP) with neXtProt and GENCODE databases, two missing proteins such as activity-regulated cytoskeleton-associated protein (ARC, Chr 8) and glutamate receptor ionotropic, kainite 5 (GRIK5, Chr 19) were additionally identified with two or more unique peptides from human brain tissues. Additionally, by applying the pipeline to human brain related data sets such as cortex (PXD000067 and PXD000561), spinal cord, and fetal brain (PXD000561), seven GENCODE ASVs such as ACTN4-012 (Chr.19), DPYSL2-005 (Chr.8), MPRIP-003 (Chr.17), NCAM1-013 (Chr.11), EPB41L1-017 (Chr.20), AGAP1-004 (Chr.2), and CPNE5-005 (Chr.6) were identified from two or more data sets. The identified peptides of GENCODE ASVs were mapped onto novel exon insertions, alternative translations at 5'-untranslated region, or novel protein coding sequence. Applying the pipeline to male reproductive organ related data sets, 52 GENCODE ASVs were identified from two testis (PXD000561 and PXD002179) and a spermatozoa (PXD003947) data sets. Four out of 52 GENCODE ASVs such as RAB11FIP5-008 (Chr. 2), RP13-347D8.7-001 (Chr. X), PRDX4-002 (Chr. X), and RP11-666A8.13-001 (Chr. 17) were identified in all of the three samples.

Some promoters were isolated and characterized from the genome of Leuconostoc mesenteroides SY2, an isolate from kimchi, a Korean traditional fermented vegetable. Chromosomal DNA of L. mesenteroides SY2 was digested with Sau3AI and ligated with BamHI-cut pBV5030, a promoter screening vector containing a promoterless cat-86. Among E. coli transformants (TFs) resistant against Cm (chloramphenicol), 17 were able to grow in the presence of 1,000 μg/ml Cm and their inserts were sequenced. Transcription start sites were examined for three putative promoters (P04C, P25C, and P33C) by primer extension. Four putative promoters were inserted upstream of a promoterless α-amylase reporter gene in pJY15α. α-Amylase activities of E. coli TFs containing pJY15α (control, no promoter), pJY03α (pJY15α with P03C), pJY04α (with P04C), pJY25α (with P25C), and pJY33α (with P33C) were 66.9, 78.7, 122.1, 70.8, and 99.3 U, respectively. Cells harboring pJY04α showed 1.8 times higher activity than the control. Some promoters characterized in this study might be useful for construction of foodgrade expression vectors for Leuconostoc sp. and related lactic acid bacteria.

In total, 170 carrot lines developed in Korea were screened for resistance to Meloidogyne incognita race 1 to select parental genetic resources useful for the development of nematode-resistant carrot cultivars. Using the gall index (GI), gall formation was examined on carrot roots inoculated with approximately 1,000 second-stage juveniles of the nematode 7 weeks after inoculation. Sixty-one carrot lines were resistant (GI ≤ 1.0), while the other 109 were susceptible (GI > 1.0) with coefficient of variance (CV) of GI for total carrot lines 0.68, indicating low-variation of GI within the lines examined. The histopathological responses of two carrot plants from resistant and susceptible lines were examined after nematode infection. In susceptible carrots, giant cells formed with no discernible necrosis around the infecting nematodes. In the resistant carrot line, however, no giant cells formed, although modified cells were observed with extensive formation of necrotic layers through their middle lamella and around the infecting nematodes. This suggested that these structural modifications were related to hypersensitive responses governed by the expression of true resistance genes. Therefore, the Korean carrot lines resistant to the nematode infection are potential genetic resources for the development of quality carrot cultivars resistant to M. incognita race 1.

This study was conducted to investigate the effect of sandplay therapy for couples on the self-differentiation, and heterosexu al relationship satisfaction, of couples who are college students’. The subjects were four couples who are college students livin g in Seoul with low self-differentiation and heterosexual relatio nship satisfaction. I carried out sandplay therapy on four coup les for once a week for 10 weeks, and each session was 70 minutes long. In this study, the self-differentiation scale was modified for Korea on the basis of Bowen’s family systems theory (1976), and it was used as a research tool. Yu (2000) and Lee (2000) made a scale to measure heterosexual relationship satisfaction after they adjusted the Korea Marital Satisfaction Inventory (K -MSI). The scale was also used in order to compare pre-therapy and post-therapy traits and identify any changes. In addition, the group’s behavior and interac tions were observed and recorde d in each session throughout the process of sandplay therapy with the couples, and the behanior and interactions were analyzed along with the change flow in the sand trays.The study’s outcomes are summed up as follows. First, sandplay therapy with couples advances the level of self-differentiation in college students. Second, sandplay therapy with couples impr oves the heterosexual relationship satisfaction on college stud ents. Third, the trays used in sandplay therapy with couples showed positive changes as the therapy progressed. As the sessions continued, negative factors in the trays decreased and the trays converged into one. As the sessions proceeded, it was observe d that the couples became closer, felt more comfortable with each other, compromised and reflected on their relationship.The study’s results show that sandplay therapy with couples inc reased the level of self-differentiation in college students and heterosexual relationship satisfaction, thus proving the th erapy’s effectiveness.

LAB were isolated from makgeolli locally produced around Jinju, Gyeongnam, S. Korea during spring of 2011. Randomly selected 11 isolates from MRS agar plates were identified first by API CHL 50 kits and then 16S rRNA gene sequencing. All 11 isolates were identified as Lactobacillus plantarum. Among them, ST4 grew in MRS broth with ethanol up to 10%, showing the highest alcohol resistance. L. plantarum ST4 was moderately resistant against acid and bile salts. When cellular proteins of L. plantarum ST4 under ethanol stress were analyzed by two-dimensional gel electrophoresis (2DE), the intensities of 6 spots increased, whereas 22 spots decreased at least 2-fold. Those 28 spots were identified by peptide mass fingerprinting (PMF). FusA2 (elongation factor G) increased 18.8-fold (6% ethanol) compared with control. Other proteins were AtpD (ATP synthase subunit beta), DnaK, GroEL, Tuf (elongation factor Tu), and Npr2 (NADH peroxidase), respectively. Among the 22 proteins decreased in intensities, lactate dehydrogenases (LdhD and LdhL1) were included.

As environmental concerns over managing seaports are gaining its importance, evaluating the greenness of the port accordingly draws a serious academic and research attention. However, the lack of the research aptly dealing with this particular problem attributes to the complexity of the problems involving various quantitative and qualitative factors, combined with the appropriateness of an evaluation structure. Therefore, this paper attempts to make contribution to the literature (1) by identifying the factors and structure of analysis by making use a factor analysis (FA); (2) by utilizing experts’ knowledge in solving the complex problems which are in nature ambiguous and unclear by adopting the fuzzy method; and (3) by ranking the targeted seaports and suggesting the pilot tail of methodology for evaluation of greenness of seaports. The results of the empirical research presented the rankings of Korean five trade ports for their greenness.

Three Weissella confusa and five Weissella cibaria strains were previously isolated from human faeces and their potential as probiotics was examined in this work. Resistance to low pHs (pH 2.0 and 3.0) and 0.3% bile salt were examined. Enzyme activities, susceptibilities to heat treatment and various antibiotics, and adhesion capacities to Caco-2 cells were also examined. All Weissella strains were killed when exposed to pH 2.0 for 2 h but survived at pH 3.0 with different survival ratios. W. confusa 31 survived best (20.2%) and W. confusa 31 was also quite resistant against 0.3% bile salt (128.8%). All strains except one grew well at temperature between 15 and 45 °C and all strains grew in the presence of 6.5% NaCl. W. confusa 20 showed the highest β-galactosidase activity (527.3 ± 23.66 unit/mg protein) and W. cibaria 31 had the highest β-glucosidase activity (115.12 ± 5.3 unit/mg protein) in MRS broth. All strains adhered to Caco-2 cells better than Lactobacillus rhamnosus GG and W. confusa 20 was the best adhesive strain (85 CFU/cell). These results show that some strains such as W. confusa 31 and W. confusa 20 are fully qualified as probiotics and deserve further application studies.

The α-amylase gene, amyL, from Bacillus licheniformis was expressed in Lactobacillus brevis 2.14 and Escherichia coli DH5α using two different shuttle vectors, pCW4 and pSJE. E. coli transformants (TFs) harboring either pCW4Tα or pSJETα produced active α-amylase but L. brevis TFs did not, as determined by enzyme assays and zymography. But amyL transcripts were synthesized in L. brevis TFs. In terms of plasmid stability, pSJE, a theta-type replicon, was more stable than pCW4, an RCR (rolling circle replication) plasmid, in L. brevis without antibiotic selection.

This study suggests for the first time that 1) IGF-I, IGFBP-4, and -5 alone and IGF-I+IGFBP-5 mixture stimulated hair cell survival and prevented neomycin-induced hair cell loss in the sensory epithelial culture of mouse utricles, 2) When administered together, IGFBP-4 diminished the effect of IGF-I, 3) In P3-5 mice utricle, IGF-I, IGFBP-4, and IGFBP-5 are expressed in the cytoplasm of hair cells. And Insulin/IGF-I Receptor is expressed in the nucleus of hair cells.Several growth factors have been demonstrated to protect auditory sensory cells in vitro and in vivo from aminoglycoside toxicity. IGF-I is one of the most well-known mitogenic and protective substance working in the inner ear. However, there are no reports available regarding the function of IGFBPs in the inner ear. In the present study, the effects of IGFBP-4 and -5 on hair cell survival were investigated in mouse utriclular organ cultures.The amount of cellular damage and cell viability in vestibular organs were assessed by counting hair cells stained with a rhodamine-phalloidin probe. The expressions of IGFBP-4, IGFBP-5, IGF-IR, and IGF-I were localized by immunohistochemistry.When treated with IGF-I, IGFBP-4, or IGFBP-5 for 24 h, explant culture showed hair cell survival rates of 136+/-18%, 140+/-15%, and 133+/-6%, respectively, compared to controls. Neomycin (1 mM) induced hair cell loss resulted in 45+/-17% of hair cell survival. However, pre-treatment of IGF-I, IGFBP-4, or -5 before neomycin insult showed survival rates of 113+/-14%, 98+/-8%, and 73+/-24%, respectively. Similar to IGF-I, IGFBP-4 and IGFBP-5 were significantly protective. IGFBP-4 and -5 immunoreactivities were observed in the cytoplasm of normal explanted vestibular hair cells as well as in the P3 mouse utricular hair cells in vivo.

The dynamics of actin filament protein is crucial for various physiological processes of the cells. Among the proteins correlating with actin dynamics, a novel 67-kDa WD40 repeat protein 1 (WDR1) was the vertebrate homologue of actin-interacting protein 1 (Aip1). Even though previous studies have provided the clues on the function of WDR1 in specific organs under pathological conditions, the exact subcellular localization of WDR1 is not known. Therefore, in the present study, we undertook to determine the distribution of WDR1 within PC12 pheochromocytoma cells (PC12 cells) using light and electron microscopic techniques. Double immunocytochemistry clearly showed that WDR1 immunoreactivities (IRs) were co-localized with anti-actin antibody, suggesting the involvement of WDR1 in actin dynamics. WDR1 immunoreactivities (IRs) in PC12 cells showed different distribution patterns as nerve growth factor (NGF) concentrations varied. During active proliferation, the distribution of WDR1 IRs seemed to be similar to those found in cortical actin patches, whereas WDR1 IR was observed in cytoplasmic actin cables after PC12 cells were induced to differentiate by treating with NGF. Though further studies are necessary to determine the function of WDR1, the current data represents a first step towards the in vitro study of WDR1 protein.

Objective: The present study was conducted to examine the effect of epidermal growth factor (EGF) on Pi uptake and its related signal pathways in the primary cultured renal proximal tubule cells (PTCs). Results: EGF (50 ng/ml) inhibited Pi uptake, a typical marker of Na+/phosphate cotransporter, in a time- and dose-dependent manner. EGF-induced inhibition of Pi uptake was blocked by AG1478 (an EGF receptor antagonist), genistein or herbimycin A (tyrosine kinase inhibitors) and also blocked by mepacrine (a phospholipase A2 (PLA2) inhibitor) and AACOCF3 (a cPLA2 inhibitor). EGF increased [3H]-arachidonic acid (AA) release, which was also blocked by AG1478, genistein or herbimycin. Furthermore, EGF-induced inhibition of Pi uptake was blocked by indomethacin (a cyclooxygenase inhibitor) and econazole (a cytochrome P-450 epoxygenase inhibitor), but not by NDGA (a lipoxygenase inhibitor). On the other hand, EGF-induced inhibition of Pi uptake was blocked by staurosporine, H-7, or bisindolylmaleimide I (PKC inhibitors), PD 98059 (a p44/42 MAPK inhibitor), but not by SB 203580 (a p38 MAPK inhibitor). EGF-induced increase of [3H]-AA release was blocked by PKC inhibitors and a p44/42 mitogen-activated protein kinase (MAPK) inhibitor, but not by a p38 MAPK inhibitor. In addition, a PKC inhibitor blocked EGF-induced phosphorylation of p44/42 MAPK. Conclusion: EGF inhibits Pi uptake via PKC-p44/42 MAPK-cPLA2 pathway in the PTCs.

Recent epidemiological studies have demonstrated that ginseng intake is associated with a reduced risk for environmentally related cancers. However, the effects of ginsenosides on the proliferation of renal proximal tubule cells have not yet elucidated. This study investigated the effect of total ginsenosides, protopanaxatriol (PT) saponin, and protopanaxadiol (PD) saponin fraction on epidermal growth factor (EGF)-induced renal cell proliferation and, furthermore, c-fos and c-jun gene expression. In the present study, total ginsenosides (10 -6 g/ml) completely blocked EGF-induced DNA synthesis and cell growth. In contrast, the PT and PD fractions partially blocked it. In addition, the EGF-induced increase of c-fos and c-jun gene expression was completely blocked by total ginsenosides and partially by PT and PD saponins. In conclusion, ginsenosides, in part, inhibit EGF-induced cell proliferation via decrease of c-fos and c-jun gene expression in primary cultured rabbit renal proximal tubular cells (PTCs). Abbreviations. EGF:epidermal growth factor PD:protopanaxadiol PT:protopanaxatriol PTCs:primary cultured renal proximal tubule cells

Oxidative stress has been implicated as a primary cause of renal failure in certain renal diseases. Indeed, renal proximal tubule is a very sensitive site to oxidative stress and retains functionally fully characterized transporters. It has been reported that ginsenosides have a beneficial effect on diverse diseases including oxidative stress. However, the protective effect of ginsenosides on oxidative stress has not been elucidated in renal proximal tubule cells. Thus, we examined the effect of ginsenosides on oxidative stress-induced alteration of apical transporters and its related mechanism in renal proximal tubule cells. In the present study, hydrogen peroxide (H2O2) (>10–5M) inhibited α-methyl-D-glucopyranoside uptake in a dose-dependent manner (p < 0.05). It also inhibited Pi and Na+ uptake. At a concentration of 20 µg/ml, total ginsenosides significantly reduced H2O2-induced inhibition of apical transporters. In contrast, protopanaxadiol (PD) and protopanaxatriol (PT) saponins exhibited a less preventive effect than total ginsenosides (p < 0.05). Furthermore, we examined its action mechanism. H2O2 increased lipid peroxide formation, arachidonic acid (AA) release, and Ca2+ uptake. These effects on H2O2 were significantly prevented by total ginsenosides and PD or PT sanponins. However, total ginsenosides appear to be more protective than PD and PT saponins (p < 0.05). In conclusion, ginsenosides prevented H2O2-induced inhibition of apical transporters via a decrease in oxidative stress, AA release, and Ca2+ uptake in primary cultured renal proximal tubule cells.

Three Pediococcus pentosaceus strains were isolated from jeotgals, salted and fermented Korean sea-foods, and their probiotic potentials were examined. After 2 h exposure to pH 3.0, P. pentosaceus F66 survived with the survival ratio of 32.6% followed by P. pentosaceus D56 (17.2%) and P. pentosaceus A24 (7.5%). P. pentosaceus F66 also survived better (26.6%) than P. pentosaceus A24 (13.7%) and P. pentosaceus D56 (5.8%) after 2 h exposure to 0.3% bile salts. Three strains grew slowly on MRS broth with 15% NaCl (w/v), reaching the OD 600 values of 0.4–0.8 in 36 h. They adhered to Caco-2 cells (10.9–13.9 CFU/cell) with similar degree of adherence of a positive control, Lactobacillus rhamnosus GG (12.8 ± 0.5 CFU/cell). Three strains possess some desirable enzyme activities such as β-galactosidase, α-glucosidase, β-glucosidase, and N-acetyl-β-glucosidase. From these results, P. pentosaceus F66 seems qualified as a probiotic and can be utilized for fermented foods including jeotgals.

Lactic acid bacteria (LAB) were isolated from Kimchi, a Korean traditional fermented vegetable food. LAB accumulating GABA (γ-aminobutyric acid) in the culture media were screened by TLC analysis. One isolate, GU240, produced the highest amount of GABA among the 3,000 isolates and identified as a Lactobacillus zymae strain. Glutamate decarboxylase (GAD) gene was cloned and over-expressed in E. coli BL21(DE3) using pET26b(+). The recombinant GAD was purified by using a Ni–NTA column. Its size was 53 kDa by SDS-PAGE. Maximum GAD activity was at pH 4.5 and 41 °C and the activity was dependent on pyridoxal 5′-phosphate. Km and Vmax of LzGAD were 1.7 mM and 0.01 mM/min, respectively, when glutamate was used as a substrate.

Unlike mammals, avian cochlear hair cells can regenerate after acoustic overstimulation. The WDR1 gene is one of the genes suspected to play an important role in this difference. In an earlier study, we found that the WDR1 gene is over-expressed in the chick cochlea after acoustic overstimulation. The aim of this study was to compare the expression of WDR1 before and after acoustic overstimulation in the chick vestibule. Seven-day-old chicks were divided into three groups: normal group, damage group, and regeneration group. The damage and regeneration group was exposed to 120 dB SPL white noise for 5–6 hours. The damage group was euthanized shortly after the impulse, but the regeneration group was allowed to recover for 2 days. The utricle, saccule, and the three ampullae of each semicircular canal were dissected and immunohistochemically stained with anti-WD40 repeat protein 1 antibody. For quantitative analysis, immunoreactive densities were measured and quantitative real-time RT PCR was performed. WD40 repeat protein 1 expression was elevated in all the semicircular canals and utricle, two days after an acoustic overstimulation (P = 0.001). WDR1 mRNA expression was 1.34 times higher in the regeneration group compared to the normal group, but it was not statistically significant. Exceptionally, WD40 repeat protein 1 expression did not increase in the saccule of the regeneration group. Elevated WDR1 expression in the avian vestibule may have a role in the hair cell regenerating ability as in the avian cochlea. A similar mechanism of hair cell regeneration may exist in the avian cochlea and vestibule.