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Sigumjang is a traditional Korean fermented barley bran consumed in the Gyungsang-do area. In this study, pork loin was marinated with sigumjang, and its physicochemical and textural properties were investigated. Sigumjang-marinated pork loin (SMPL) displayed an attractive yellowish-brown overall color. SMPL’s pH marginally increased during storage and finally equilibrated to the pH of sigumjang, 5.5, on day 14. The amino-type nitrogen content of SMPL increased linearly during the marination period. Due to the lower extractability and higher pH compared to the control, the volume of myofibrillar fragments in SMPL decreased during the marination period. SMPL showed an increase in free amino acids related to umami (Asp and Glu) and sweet (Ser, Thr, Ala and Gly) tastes compared to the control. These may indicate improvement of sensory qualities of SMPL. The results also provide valuable information for the use of sigumjang in the development of novel meat products.

Fermented barley bran (FBB) using Bacillus spp. is the main ingredient of sigumjang, a traditional fermented food in South Korea. The aim of this study was to evaluate the gastro-protective effects of β-glucan-enriched FBB extracts in an experimental mouse model of the EtOH-induced gastric ulcer. The gastro-protective activity of FBB was determined using histopathological inspection, level of EtOH absorption from the gastrointestinal-tract and measurement of pro-inflammatory cytokine. Pretreatment of FBB showed less gastrointestinal bleeding and reduced EtOH absorption into the bloodstream with the gastric mucosa. Furthermore, FBB effectively reduced the EtOH-induced mRNA expression of tumor necrosis factor α in the gastric mucosa and its level in the blood. These results showed that FBB can prevent gastric mucosal damage induced by acute EtOH-administration in mice by reducing absorption of EtOH by gastric mucosa along with decreasing the inflammatory response. Altogether, this data provides useful information on how β-glucan-enriched food modulates the EtOH-induced gastric mucosal damage, and thus, it could lead to the development of formulations for protecting gastric mucosal injury with acute alcohol intake.

Sigumjang is a traditional Korean fermented barley bran consumed in the Gyungsang-do area. In this study, pork loin was marinated with sigumjang, and its physicochemical and textural properties were investigated. Sigumjang-marinated pork loin (SMPL) displayed an attractive yellowish-brown overall color. SMPL’s pH marginally increased during storage and finally equilibrated to the pH of sigumjang, 5.5, on day 14. The amino-type nitrogen content of SMPL increased linearly during the marination period. Due to the lower extractability and higher pH compared to the control, the volume of myofibrillar fragments in SMPL decreased during the marination period. SMPL showed an increase in free amino acids related to umami (Asp and Glu) and sweet (Ser, Thr, Ala and Gly) tastes compared to the control. These may indicate improvement of sensory qualities of SMPL. The results also provide valuable information for the use of sigumjang in the development of novel meat products.

Hazardous metals leaching experiment was carried out in accordance with various usage environments for camping cooking utensils distributed in the market. There was a significant difference in the degree of migration for lead, arsenic, cadmium and nickel defending on the solvent and how to use, although they were all appropriate for criteria. In general, the migrated amount of aluminum was increased in acidic condition, and the migrated amount of arsenic was increased in salty condition. Physical scratches increased the overall release of hazardous metals from the portable pots and pans for camping in all solvents. Especially, in 0.5% citric acid solution, cadmium was migrated by physical scratch in stainless steel and hard aluminum pots and pans. The longer the leaching time, the higher the migration of aluminum in acid condition and arsenic in basic condition. From these results, it is desirable to use the cooking utensil for camping without being exposed to strong acidic or basic solution and scratches in order to reduce the migration of hazardous metals from them.

Sigumjang prepared from fermented barley bran is a traditional fermented food found only in the Gyeongsang-do area of South Korea. There have been no studies reported to date despite the potential bioactivities of sigumjang. In this study, the anti-obesity activities of sigumjang extracts (SEs) during 3T3-L1 differentiation into adipocytes were investigated. SEs inhibited adipocyte differentiation by suppressing the CCAAT/enhancer binding protein-β and sterol regulatory element binding protein-1c expression in the early stage of differentiation, followed by the suppression of the peroxisome proliferator-activated receptor-γ, CCAAT/enhancer binding protein-α, and adiponectin. These changes in adipogenic markers induced inhibition of lipogenesis via down-regulation of mainly fatty acid synthase, acetyl-CoA carboxylase, fatty acid binding protein 4, and perilipin. These results were more significant in the extract of sigumjang fermented with isolated Bacillus amyloliquefaciens MFST compared to naturally fermented sigumjang group. SEs can be considered as a useful material for developing food with health benefits and anti-obesity properties.

Background: We conducted this experimental study to examine whether human adipose-derived stem cells (ADSCs) are effective in achieving a recovery of damaged renal tubular epithelial cells in an animal model of cisplatin-induced acute kidney injury using rats. Methods: To examine the in vitro effects of ADSCs in improving nephrotoxicity, we treated mouse renal tubular epithelial cells with both ADSCs and cisplatin mouse renal tubular epithelial cells. And we equally divided 30 male white Sprague-Dawley (SD) rats into the three groups: the control group (intraperitoneal injection of a sterile saline), the cisplatin group (intraperitoneal injection of cisplatin) and the ADSC group (intraperitoneal injection of cisplatin and the hADSC via the caudal vein). At five days after the treatment with cisplatin, serum levels of blood urine nitrogen (BUN) and creatinine were measured from each SD rat. We performed histopathologic examinations of tissue samples obtained from the kidney. Results: The degree of the expression of TNF-α and that of Bcl-2 were significantly higher and lower respectively, in cisplatin group (P

Synergistic fermentation between bacteria and fungi has great potential in many industrial fields. Here, we investigated how bacterial-fungal fermentation can promote the bioactivity of edible-fungus. The submerged Lactobacillus plantarum culture actively produced acids incorporated with polysaccharides from β-glucan-rich Pleurotus eryngii as a substrate for fermentation that resulted in a low pH of submerged culture. Fermented P. eryngii with L. plantarum showed enhanced bioactivities, including anti-bacterial, antioxidant, and anti-allergic activities compared to hot water extract of P. eryngii. This result suggests that a synergistic fermentation between bacteria and fungi can apply to functional food making.

Background Proteinuria is typically accompanied by structural and compositional changes of the foot processes and of the slit diaphragms between podocytes. CD2-associated protein (CD2AP) in podocytes serves as an adaptor protein binding to nephrin and podocin, anchoring these slit diaphragm proteins to actin filaments of podocyte cytoskeleton and sending signals inward or outward. Methods In the present study, we prepared streptozotocin-induced diabetic renal tissues and cultured podocytes in diabetic conditions to investigate podocyte phenotypical changes, including quantitative and distributional changes of CD2AP protein and search for the signalling mechanisms in diabetic conditions. We prepared cultured rat glomerular epithelial cells and mouse podocytes to study how high glucose and advanced glycosylation end products (AGE) induce phenotypical changes of cultured podocyte, under (1) normal glucose (5 mM, = control), (2) high glucose (30 mM), (3) AGE-added or (4) high glucose plus AGE-added conditions. Results According to diabetic duration, density of CD2AP in renal tissue of experimental diabetic nephropathy became conglomerulated and diminished. In cultured podocytes, CD2AP co-localized with nephrin and zonula occludens-1 by confocal imaging. High glucose and high glucose plus AGE induced the relocalization and concentration of CD2AP at internal cytoplasmic and perinuclear areas of podocytes. High glucose plus AGE-added condition also decreased CD2AP protein amount and its mRNA expression compared with normal glucose or osmotic control conditions. In addition, LY294002, a phosphoinositide 3-kinase inhibitor, prevented the quantitative and distributional changes of CD2AP induced by high glucose and AGE. Conclusions These findings suggest that diabetic conditions induce the phenotypical changes of podocyte CD2AP possibly via phosphoinositide 3-kinase/Akt signalling. Copyright © 2014 John Wiley & Sons, Ltd.

Although the pharmacological action of red ginseng is mostly generated by ginsenosides, contents of ginsenosides in manufactured extracts are still varying according to processing and storage conditions. Rg1, Rb1, and Rh1 levels significantly decreased overtime under pH adjustment and thermal treatment during storage, and these changes were exacerbated at lower pH and higher storage temperature. However, Rg3 level showed the opposite pattern compared to other ginsenosides. Rg3 level linearly increased at higher temperature and pH while significantly decreased at pH 2. Furthermore, Rg3 level constantly increased during storage. This is the first combined study on the effects of manufacturing and storage conditions on ginsenoside contents of red-ginseng-extract products. To minimize loss of major marker and bioactive compounds of red ginseng products during manufacturing processes and storage, it is recommended that red-ginseng-extracts be maintained at pH 6–8, sterilized at below 105 °C, and stored at below 25 °C.

The effects of Jerusalem artichoke (JA) on hypoglycemia and hepatoprotection in streptozotocin (STZ)-induced diabetic rats were investigated. Water extract of Jerusalem artichoke (JAE) reversed the increase in serum glucose levels in STZ-induced diabetic rats. Further, JAE treatment remarkably prevented elevation of serum aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, and γ-glutamyl transpeptidase levels in STZ-induced diabetic rats. Treatment with JAE also improved lipid profiles damaged by STZ in the liver by reducing serum levels of triglycerides and total cholesterol. These results suggest that oral administration of JAE may have a potential benefit of ameliorating diabetic symptoms by improving liver damage caused by STZ. This study gives useful information for understanding the anti-diabetic effect of JA.

Pleurotus ostreatus (PO), which is also called the oyster mushroom, is the secondly most popular cultivated mushroom in the world and has been shown to display antioxidant activity. In this study, the hepatic protective effects of extracts of PO cultured in wheat (POW) on carbon tetrachloride (CCl4)-induced liver injury in rats were evaluated by analyzing blood markers of liver injury. Sprague-Dawley rats were pretreated with POW extracts (0, 0.5, and 1.0 g/kg) orally for 14 days prior to the administration of CCl4 for 3 days. Marked evaluation of alanine and aspartate aminotransferases, lactate dehydrogenase, alkaline phosphatase, and γ-glutamyltransferase were observed in the plasma from control rats after CCl4 treatment. This increased liver injury markers were significantly reduced in a dose-dependent manner by pretreatment with the POW extract, suggesting that POW prevented acute liver damage in CCl4-intoxicated rats by suppressing cellular leakage and loss of the functional integrity of cell membranes in the liver. POW extracts could also improve lipid profiles damaged caused by CCl4 in liver through a reducing plasma triglyceride and total cholesterol and a recovering plasma HDL-cholesterol. These results suggest that the extracts of POW ameliorate hepatotoxicity induced by CCl4 in the rat model.

Changes in extractability and bioactivity of fermented rice bran (FRB) were investigated using high hydrostatic pressure (HHP) treatment alone (HFRB) and in combination with complex enzymes (CHFRB). The CHFRB treatment resulted in increased contents of total sugars and free amino acids compared to both the HFRB and control groups, which had undergone hot-water extraction. A high performance liquid chromatography analysis showed that CHFRB significantly increased the volume of ferulic acid, although there was no significant difference in total phenolic content among the groups. In bioactivity analyses, the CHFRB extract showed significant increases for inhibiting linoleic acid oxidation and angiotensin-converting enzyme activity compared to those in the other groups. Based on these results, HHP treatment combined with complex enzymes efficiently increased the extractability of ferulic acid and free amino acids, which improved bioactivity. These results provide useful information for using FRB as an ingredient in functional foods and for exploiting HHP treatment combined with specific enzymes to improve the extractability and bioactivity of a raw material. Industrial relevance This study will give a novel technique for using rice bran, a byproduct of the rice milling process, which contains various bioactive materials since most of them have been discarded uselessly. Our results provide useful information for using rice bran (fermented) as an ingredient source in functional foods and for exploiting HHP treatment combined with specific enzymes to improve the extractability and bioactivity of the raw material.

Context Ligularia fischeri (Ledebour) Turcz. (Compositae) has been used as a leafy vegetable and in traditional medicine to treat hepatic disorder in East Asia. Objective The present study explores the antioxidant activity of LF aqueous extract on EtOH-induced oxidative stress accompanied by hepatotoxicity both in vitro and in vivo. Materials and methods In vitro study using the mouse liver NCTC-1469 cell line was conducted to estimate the cytotoxicity as well as the inhibitory effect of LF extract against alcohol-treated cell damage. In vivo study used an alcohol-fed Wister rat model orally administered EtOH (3.95 g/kg of body weight/d) with or without LF extract (100 or 200 mg/kg body weight) for 6 weeks. Serum and liver tissue were collected to evaluate hepatic injury and antioxidant-related enzyme activity. Results The EC50 value for the DPPH radical scavenging capacity of LF extract was 451.5 μg/mL, whereas the IC50 value of LF extract in terms of EtOH-induced reactive oxygen species (ROS) generation was 98.3 μg/mL without cell cytotoxicity. LF extract (200 mg/kg body weight) significantly reduced the triglyceride content of serum (33%) as well as hepatic lipid peroxidation (36%), whereas SOD activity was elevated three-fold. LF extract suppressed expression of CYP2E1 and TNF-α, and attenuated alcohol-induced abnormal morphological changes. Discussion and conclusion LF extract attenuated liver damage induced by alcoholic oxidative stress through inhibition of ROS generation, down-regulation of CYP2E1, and activation of hepatic antioxidative enzymes. Homeostasis of the antioxidative defence system in the liver by LF extract mitigated hepatic disorder following chronic alcohol intake.

Plant-derived polyhenols inhibit cancer cell proliferation and induce apoptosis. Recently, prenylflavonoids and alkyl-phloroacetophenones have been reported for their in vitro antitumor activity. In the present study, we examined the cytotoxic activity of prenyl (3-PAP) and geranyl (3-GAP) derivatives of phloroacetophenone, and xanthohumol (XN), a prenyl-chalcone, in human breast cancer (MCF-7) and human sarcoma (HT1080) cell lines in vitro. 3-GAP showed the strongest cytotoxicity in these cell lines with IC(50) values of less than 10 µM. In addition, we report that 3-GAP is a more potent anti-cancer agent for breast cancer than XN which is a well-known anticancer flavonoid. Moreover, 3-GAP did not induce cytotoxicity in the normal cell line, TCMK-1, whereas 3-PAP and XN significantly reduced TCMK-1 cell viability. In 3-GAP-treated MCF-7 cells, nuclear accumulation and transcriptional activity of p53 were increased. In addition, pro-apoptotic Bax but not B-cell lymphoma 2 (Bcl-2) expression was increased by 3-GAP. In accordance with the Bax increase, 3-GAP induced mitochondrial cytochrome c release and activated caspase-9, an initiator of the caspase cascade. In the MCF-7 cell line which does not express caspase-3, activation of caspase-7, a member of the caspase-3 subfamily, was increased by 3-GAP. The present results indicate that synthetic 3-GAP is a safe and effective anti-cancer agent, and the Bax-mediated mitochondrial pathway is the main apoptosis signaling pathway of 3-GAP in MCF-7 cells.

Investigation of the optimum conditions for the combined application of Saccharomyces sp. (yeast) and Leuconostoc sp. (lactic acid bacteria, LAB) isolated in a previous study to the development of novel sourdough was carried out by response surface analysis. First, the cell growth conditions were analyzed. LAB showed good proliferation under conditions of 30–35°C, low pH, and high acidity, whereas the growth of yeast was inhibited. The growth of yeast was optimum at 25°C for 24 h. Based on these results, analysis of sourdough was carried out by varying the LAB population, temperature, and time after fixing the number of yeast. It was determined by response surface analysis that the optimal conditions for fermentation are LAB population of 105 CFU/mL, temperature of 25°C, and reaction time of 24 h. From these results, the growth of LAB should be constantly maintained, and an appropriate pH that does not inhibit the growth of yeast due to the presence of generated organic acid is required to allow for the unique properties of sourdough. This study could give useful information for the development of novel sourdough.

In this study, the effects of water extracts of fermented rice bran (FRBE) on immediate-type allergic reactions in vitro (RBL-2H3 cell line) and in vivo [ovalbumin (OVA) sensitized mice model] were investigated. FRBE inhibited β-hexosaminidase release from RBL-2H3 cells activated by anti-dinitrophenyl (DNP) IgE while FRBE pretreatment profoundly inhibited the systemic allergic reaction induced by compound 48/80. Moreover, FRBE had a significant inhibitory effect on the mRNA expression of anti-DNP IgE-induced tumor necrosis factor (TNF)-α, interferon (IFN)-γ, interleukin (IL)-4, IL-6, and IL-10, all of which mediate inflammatory reactions triggered by allergic reaction. To determine the mRNA levels of cytokines in an OVA-sensitized mice model after FRBE treatment, spleen cells were harvested with several concentrations of FRBE. The FRBE-treated group showed decreased levels of TNF-α, IFN-γ, IL-6, and IL-10. These results suggest that FRBE suppressed allergic and inflammatory reactions through inhibition of degranulation, histamine release, and pro-inflammatory cytokine production from mast cells.

A 90-d trial was conducted to determine the influence of additives such as pegmatite (Peg), precious stone (PS), grapepomace extracts (GPEx) and complexes of these additives (Peg + GPEx and PS + GPEx) via evaluation of meat qualityincluding sensory properties in 90 d old finishing pigs. There were no significant differences in the approximate composi-tion and physiochemical characteristics, including meat color, among the treated groups. However, the Peg treated groupwas found to have a higher water holding capacity than the other groups. The shear-force was evaluated as an indicator ofhardness and found to be lower in the Peg and GPEx treated groups, and this tendency was greater when the complex of Pegand GPEx was supplied. No significant differences were observed among groups following analysis of the free amino acidcomposition and fatty acid (FA) composition. Sensory evaluation of the boiled loins showed significantly (p

The prevalence of food allergies was investigated using questionnaires with 300 subjects whose ages ranged from 19 to 24 years old and the causative food allergens was analyzed using immunological analysis with serum of the subjects who answered that they have/had food allergy. The questionnaire showed that 11.33% of subjects have/had experience of food hypersensitivity, where the main causative foods were fish, beef, chicken, milk, egg, and pork in order. The meat allergy shared 4.65% (2.33% for beef, 1.66% for chicken, 0.66% for pork) in the prevalence of food allergies. The causative beef allergens were investigated with the serum of 6 subjects who have had beef allergy. Western blots were carried out with the serum of P6 subject who showed a positive reaction to beef extract in ELISA. The two specific bands were detected in beef extract on the PVDF membrane, and no band was detected in extracts of pork and chicken. A calculation of the distance of migration by SDS-PAGE enabled the molecular masses of the two bands to be estimated as 67kDa and 31kDa, respectively. The 67kDa was revealed as bovine serum albumin (BSA) which is one of the important beef allergens as reported previously though an analysis of the N-terminal amino acid sequence. However we could not identify the sequence of 31kDa, probably because they comprised several subunits and were modified proteins such as glycoprotein that were unlikely to be easily degraded by the Edman method. The 31kDa band were dyed with the PAS (periodic acid-schiff reagent), suggesting that it might be a glycoprotein. These results suggested that the 31kDa might be considered as a novel potential beef allergen which is not reported previously, although further studies are needed.

This study was carried out to evaluate the correlation between shear force determined by tenderness analyzer and grade decision factors consisting of yield grade (carcass weight, ribeye area, backfat thickness) and quality grade (meat color, fat color, marbling, maturity, texture) in a total of 200 Hanwoo cows. Results showed that there was a negative correlation between shear force and yield grade (r

This study was conducted to evaluate the breadmaking characteristics of white bread when rice bran (RB), fermented rice bran (FRB) and rice bran oil (RBO) were added to the dough. Addition of RB to dough generally showed a decrease of the bread volume and increase of the bread weight which is related to the comparative decrease of gluten in the dough. Addition of RB also showed a decrease of lightness with yellowish color of RB and showed some rheological changes in dough. However, up to 5% RB addition, the dough showed a similar property of breadmaking with giving functional benefits compared to the control. Moreover 5% FRB addition to the dough improved the taste, flavor, and texture important to quality of bread. Substitutional use of RBO for shortening in dough result in comparative decrease of volume of dough. Addition of yellowish RBO influenced the increasing yellowness score, hardness, gumminess and brittleness, which is disadvantageous for the quality of white bread. However up to 50% RBO substitution for shortening made little rheological differences compared to the control. The 50% RBO substitution for shortening increased the score of flavor in sensory test even if there was an unfavorable quality point of white bread caused by a decreased volume and an increased hardness. From these results it could be suggested that the proper use of RB, FRB and RBO in breadmaking could improve the taste and flavor, giving beneficial function originated from rice bran to white bread.

This study shows the effects of L-carnitine treatment on cell proliferation with hepa1c1c7 mouse cancer cells and NCTC 1469 normal cells. In an MTT assay, L-carnitine increased the number of dead hepa1c1c7 cells, while there was no difference in the number of NCTC 1469 cells. mRNA and protein levels of TNF-alpha, Fas, and caspase-8, which are closely related to cell apoptosis by a death ligand/receptor-dependent apoptosis pathway, were increased by L-carnitine treatment. In addition, L-carnitine treatment regulated mitochondria-dependent apoptosis pathways by inducing the up-regulation of caspase-9 and caspase-3 and the down-regulation of Bcl-2 in hepa1c1c 7 cells. Taken together, the findings of this study have demonstrated that L-carnitine could induce apoptosis in hepa1c1c7 cells by regulating Fas ligands and inhibiting the expression of Bcl-2. These results suggest that L: -carnitine treatment could be related to both a mitochondrion-dependent and a death ligand/receptor-dependent apoptosis pathway in hepa1c1c7 cells. Our results could give information for understanding the L-carnitine-induced apoptosis mechanism in some cancer cells.

Although angiotensin II (Ang II) type 1 receptor antagonist ameliorates proteinuria, its pharmacological mechanism and the differential roles of Ang II type 1 receptor (AT1R) and type 2 receptor (AT2R) are not well understood. We analyzed the effect of Ang II type 1 receptor antagonist on proteinuria caused by antibody against nephrin, a functional molecule of glomerular slit diaphragm and dysfunction of which is involved in the development of proteinuria in several glomerular diseases. We show here that AT1R antagonist ameliorated proteinuria by preventing a reduction in the functional molecules of the slit diaphragm. We also analyzed the role of AT1R- or AT2R-mediated actions on the expression of the slit diaphragm molecules in an in vivo study of normal rat and in an in vitro study of cultured podocytes. AT1R-mediated action hampered the mRNA expression of the slit diaphragm molecules, whereas AT2R-mediated action enhanced it. These findings indicate that Ang II receptor subtypes play opposite roles in regulating the barrier function of glomerular capillary wall and that the enhancement of AT2R stimulation may serve as a potential therapeutic strategy for proteinuria.

Synaptic vesicle protein 2B (SV2B) was identified by the subtraction hybridization technique as a molecule of which mRNA expression was decreased in puromycin aminonucleoside (PAN) nephropathy by glomerular cDNA subtraction assay. The expression of SV2B was detected in glomerular lysate with Western blot analysis. Dual-labeling immunofluorescence studies with glomerular cell markers demonstrated that SV2B is expressed in glomerular visceral epithelial cells (podocytes). The expression of SV2B is detected also in cultured podocyte and in human kidney section as podocytic pattern. The decrease of SV2B mRNA was already detected before the onset of proteinuria in PAN nephropathy. The mRNA expression of SV2B clearly is altered not only in PAN nephropathy but also in another proteinuric state that is caused by an antibody against nephrin, a functional molecule of the slit diaphragm. The decreased intensity in SV2B staining was already detected before the peak of proteinuria in both models with immunofluorescence study. A reduced amount of SV2B was detected in both models also with Western blot analysis. CD2AP, another functional molecule of the slit diaphragm, was observed in cytoplasm, including the processes area of the cultured podocyte, and when the podocyte was treated with small interfering RNA for SV2B, CD2AP staining at the process area was not detected. These results suggest that SV2B is a functional molecule of podocyte, and SV2B may play a role in the expression and proper localization of CD2AP.

Exocrinopathy and pancreatitis-like injury were developed in C57BL/6 (B6) mice infected with LP-BM5 murine leukemia virus, which is known to induce murine acquired immunodeficiency syndrome (MAIDS). The role of chemokines, especially CXCL10/interferon (IFN)-γ-inducible protein 10 (IP-10), a chemokine to attract CXCR3+T helper 1-type CD4+T cells, has not been investigated thoroughly in the pathogenesis of pancreatitis. B6 mice were inoculated intraperitoneally with LP-BM5 and then injected every week with either an antibody against IP-10 or a control antibody. Eight weeks after infection, we analyzed the effect of IP-10 neutralization. Anti-IP-10 antibody treatment did not change the generalized lymphadenopathy and hepatosplenomegaly of mice with MAIDS. The treatment significantly reduced the number of IP-10- and CXCR3-positive cells in the mesenteric lymph nodes (mLNs) but not the phenotypes and gross numbers of cells. In contrast, IP-10 neutralization reduced the number of mononuclear cells infiltrating into the pancreas. Anti-IP-10 antibody treatment did not change the numbers of IFN-γ+and IL10+cells in the mLN but significantly reduced their numbers, especially IFN-γ+and IL-10+CD4+T cells and IFN-γ+Mac-1+cells, in the pancreas. IP-10 neutralization ameliorated the pancreatic lesions of mice with MAIDS probably by blocking the cellular infiltration of CD4+T cells and IFN-γ+Mac-1+cells into the pancreas at least at 8 wk after infection, suggesting that IP-10 and these cells might play a key role in the development of chronic autoimmune pancreatitis.

SUMMARY: Although the role of glomerular basement membrane has been emphasised as the barrier for retaining plasma proteins in the past three decades, some recent studies have demonstrated that the slit diaphragm of the glomerular epithelial cell (podocyte) is the structure likely to be the barrier in the glomerular capillary wall. Nephrin and podocin were identified as gene products mutated in Finnish-type congenital nephrotic syndrome and autosomal recessive steroid-resistant nephrotic syndrome, respectively. Nephrin s located at the outer leaflet of plasma membranes of the slit diaphragm. Podocin is reported to have an interaction with nephrin. The anti-nephrin antibody is capable of inducing massive proteinuria, which indicates that nephrin is a key functional molecule in the slit diaphragm. The expression of nephrin and podocin was reduced in glomeruli of minimal change nephrotic syndrome, which suggested that the altered expression of these molecules contributes to the development of proteinuria also in acquired diseases. Some recent studies demonstrated that CD2-associated protein (CD2AP) is also a functional molecule in the slit diaphragm, and its expression is altered in membranous nephropathy. These observations suggested that alteration of the molecular arrangement in the slit diaphragm is involved in the development of proteinuria in several kinds of glomerular diseases.

Recent investigations have revealed the importance of glomerular podocytes with its diaphragm as the major filtration barrier. Junctional adhesion molecule 4 (JAM4) has been identified as a protein that interacts with membrane-associated guanyl kinase inverted (MAGI)-1 and is reported to be expressed on podocytes. To elucidate the role of JAM4 on podocytes, we examined the expression of JAM4 and MAGI-1 in normal and two different proteinuric rat models: puromycin aminonucleoside (PAN) nephropathy and anti-nephrin antibody-induced (ANA) nephropathy, one model with and one without effacement of podocyte foot processes. JAM4 was detected by immunomicroscopy at the apical membrane of normal podocytes. JAM4 immunostaining was focally increased in the podocytes in PAN nephropathy but not in ANA nephropathy. In proteinuric podocytes, the expression of JAM4 was distinct from that of MAGI-1 or other slit diaphragm molecules such as nephrin and ZO-1. Close colocalization of JAM4 and ezrin was maintained in PAN nephropathy. By immunoelectron microscopy, the signals for JAM4 were detected at the free apical membrane of the podocytes with effaced foot processes. Studies with selective detergent extract revealed that the subcellular localization of JAM4 was altered in PAN nephropathy. Thus the altered expression of JAM4 appears to be associated with morphological changes in podocytes and can be a useful marker of injured podocytes. JAM4 may have a different role at the apical membrane besides the role as a junctional molecule and is likely associated with the unique structure of this epithelium.

IFN-inducible protein-10 (IP-10/CXCL10) is a potent chemoattractant for activated T lymphocytes and was reported recently to have several additional biologic activities. In this study, the pathophysiologic role of IP-10 in the glomerular visceral epithelial cell (podocyte) was investigated. In cultured podocytes subjected to recombinant IP-10 treatment, the expression of slit-diaphragm (SD) components nephrin and podocin clearly was heightened. Rats that had puromycin aminonucleoside nephropathy and anti-nephrin antibody-induced nephropathy and were subjected to anti-IP-10 function-blocking antibody (anti-IP-10 mAb) treatment displayed a decrease in the protein level of SD components, as well as exacerbated proteinuria. For exploration of the mechanisms of this process, the interaction between IP-10 and the cell-cycle regulatory proteins was investigated. Cultured podocytes subjected to recombinant IP-10 treatment displayed an increase in the protein level of p27(Kip1), whereas the levels of cyclins E and A decreased. The expression of IP-10 and SD components was heightened by the treatment of siRNA of cyclin A, whereas these expressions were lowered by the treatment of siRNA of p27(Kip1). Proteinuric rats subjected to anti-IP-10 mAb treatment displayed a heightened expression of cyclin A from the early phase of the disease, which indicates that the anti-IP-10 mAb treatment exacerbates podocyte injury by disturbing the cell-cycle balance. These results raise the possibility that IP-10 could become a novel therapeutic target in nephrotic syndrome and several diseases with altered cell-cycle balance.

Each of the aromatic, acidic and basic amino acid residues in HM-1 were separately substituted with alanine by site-directed mutagenesis. The mutant genes were successfully expressed in HM-1 resistant Saccharomyces cerevisiae. HM-1 gene analogues corresponding to the aromatic substitutions resulted in lower production of HM-1 analogues. In the case of the acidic amino acid residue and basic amino acid residue substitutions, some analogues were produced in the same amount as and exhibited similar killing activity to that of the wild type HM-1. But the H35A HM-1 analogue had completely lost the killing activity, and D44A, K21A, K46A, R82A, R85A and R86A HM-1 showed highly decreased killing activities. These results strongly indicate the importance of histidine-35, aspartic acid-44, lysine-21, lysine-46, and C-terminal arginine residues in HM-1 for the killing activity.

Background/Aims: Multi-glycoside from Tripterygium wilfordii Hook f. (GTW) is used for various immune and inflammatory diseases including renal diseases represented by mesangial proliferative glomerulonephritis (MsPGN) in China. However, there have been no fundamental studies on the operating mechanism of GTW on MsPGN. The aim of this study is to examine as the first step the effects of GTW on acute injurious process such as mesangial injury and proteinuria in an acute and reversible Thy.1.1 glomerulonephritis (Thy1.1GN) model and then to clarify the action mechanism of GTW at molecular level by examining its effects on various injurious factors in this model. Methods: Thy1.1 GN was induced in rats by a single intravenous injection with 500 µg of anti-Thy1.1 mAb 1-22-3. Daily oral administration of GTW and vehicle as a control was started from 3 days before injection of mAb to the day of sacrifice in each experiment. Fourteen rats were randomly divided into 2 groups, GTW-treated and vehicle-treated groups, and sacrificed on day 14 in experiment 1 or on day 7 in experiment 2 after induction of Thy1.1 GN. Proteinuria was determined on days 1, 3, 5, 7, 10 and 14 in experiment 1 or on 1, 3, 5 and 7 in experiment 2. From blood and kidneys taken at sacrifice, blood biochemical parameters, mesangial morphological changes, glomerular macrophage infiltration, and glomerular mRNA expression of cytokines were examined. Results: In experiment 1, proteinuria and mesangial matrix expansion were significantly attenuated by GTW treatment. In experiment 2, GTW treatment significantly ameliorated proteinuria, mesangial lesions and macrophage accumulation in glomerulus. In addition, it significantly reduced the glomerular expression of mRNA for PDGF, MCP-1 and IL-2. Conclusion: GTW ameliorated not only proteinuria but also mesangial alterations in Thy1.1 GN most likely by reducing expression of injurious cytokines, indicating that GTW has suppressive effects on acute inflammatory changes in glomeruli.

The sera of bovine gamma globulin (BGG) positive beef allergic patients were used in this study in order to investigate changes in IgE-specific binding activity with regard to beef extract altered by heat or high-pressure treatment. In inhibition-ELISA, the sample treated at 60 degrees C did not show any significant changes in the antigenicity of BGG, but the sample treated at 100 degrees C showed a decrease of the antigenicity. In the case of the treatment with heating at 100 degrees C, heat-coagulation occurred in the beef extract. The resulting supernatant and precipitate of the sample by centrifugation were analyzed by immunoblotting. Only the fraction of precipitate showed a specific binding activity with the sera. Based on this result, it was speculated that the persistent antigenicity found even after the treatment at 100 degrees C in inhibition-ELISA remained principally in the heat-coagulated fraction, which indicated the importance of the method of handling the heat-coagulation in heat treatment. High-pressure treatments (200 MPa-600 MPa) of beef extract did not show any significant changes in the binding with the sera.

Cereal grains are considered to be one of the most pivotal sources of dietary nutrients. Bran, which is high in fiber and contains a phytochemical-rich outer layer of cereal grains, is considered an important ingredient for reducing the risk of lifestyle-related diseases. Bran is composed of phenolics bound in dietary fibers and β-glucan, which has a wide spectrum of biological activities. To date, solid-state fermentation has emerged as a potential technology for the development of bioactive products. It is also commonly employed to increase the content of phenolic compounds in certain foods. Hence, fermented cereal grains such as bran with abundant amounts of bioactive molecules are good sources of antioxidants as well as ingredients. Fermented rice bran with yeast has been reported to have various bioactivities, and is considered as a new type of functional food ingredient containing novel metabolites.

The potential allergenic proteins in beef were investigated. The sera of ten beef-allergic patients suffering from atopic dermatitis and having a positive RAST score to beef, aged 3-18 years, were obtained from Yoshida Hospital in Japan, and five non-allergic individuals were subjected to this study. The sera of the ten patients reacted strongly to a beef extract, but not to pork and chicken extracts by both ELISA and immunoblotting. The sera of the five control subjects did not react to any of these meat extracts. Three bands having molecular masses of approximately 200 kDa, approximately 67 kDa and approximately 60 kDa were observed by immunoblotting after SDS-PAGE. Two fractions of the beef extract from a Sephadex-gel (G-200) filtration column strongly reacted with the sera of the beef-allergic patients by ELISA and immunoblotting: one fraction had the approximately 67 kDa component and the other had the approximately 200 kDa and approximately 60 kDa components. One of them (approximately 67 kDa) was confirmed to be bovine serum albumin (BSA) by an analysis of the N-terminal amino acid sequence. We could not identify the others by sequencing, but the approximately 200 kDa and approximately 60 kDa components were presumed to be glycoproteins. Bovine gamma (BGG:globulin M.W. approximately 160 kDa) is a glycoprotein and has several subunits. The beef-allergic patients showed strong reactivity to the approximately 200 kDa and approximately 60 kDa components of pure BGG by immunoblotting. Inhibition-ELISA showed that pure BGG preparations strongly inhibited the binding of sera from the beef-allergic patients to the beef extract. These results suggest that the approximately 200 kDa, approximately 67 kDa and approximately 60 kDa components in the beef extract had strong allergenicity: approximately 67 kDa was BSA, and approximately 200 kDa and approximately 60 kDa were presumably aggregated BGG and it's heavy chain, respectively.

Dongyeop Kim,1,3* Jin-Hong Park,2* Dae-Jun Kweon,2 Gi Dong Han11Department of Food Science and Technology, College of Natural Resources, Yeungnam University, Gyeongsan, Republic of Korea; 2BioHealth Convergence Center, Daegu Technopark, Daegu, Republic of Korea; 3Division of Applied Bioscience, Graduate School of Agriculture, Hokkaido University, Sapporo, Japan *These authors contributed equally to this workBackground: The aim of this study was to enhance the bioavailability of conjugated linoleic acid (CLA), which has low water solubility, using nanoemulsion technology and to evaluate the effects of its improved bioavailability as an antiobesity agent.Methods: The antiobesity effect of nanoemulsified water-soluble conjugated linoleic acid (N-CLA) was evaluated using in vitro and in vivo studies. Differentiated 3T3-L1 adipocytes were treated with CLA and N-CLA to assess their lipolytic effect. Further, to confirm the antiobesity effect of N-CLA, male Sprague-Dawley rats were randomly separated into four groups, ie, a group fed a normal diet, a group fed a high-fat diet (obesity rat model), a CLA-treated group, and an N-CLA-treated group.Results: N-CLA showed a greater lipolytic effect on differentiated 3T3-L1 adipocytes compared with normal CLA. N-CLA enhanced the release of glycerol from triglycerides, which accumulated in differentiated 3T3-L1 adipocytes. Further, N-CLA enhanced leptin secretion to an extent similar to that of orlistat, an antiobesity agent. In an animal obesity model fed a high-fat diet, N-CLA attenuated accumulation of triglycerides, total cholesterol, and low-density lipoprotein cholesterol in serum, and also significantly decreased the volume of triglycerides and cholesterol in liver tissue.Conclusion: These results indicate that N-CLA has a greater antiobesity effect than CLA as a result of its improved bioavailability.Keywords: conjugated linoleic acid, nanoemulsion, water-soluble, improved bioavailability, antiobesity

Background Proteinuria results from increased glomerular permeability and is associated with retraction and effacement of the highly specialized interdigitating podocyte foot processes. In podocytes (glomerular epithelial cells [GECs]), p130Cas localizes diffusely to the cytoplasm and connects focal adhesion proteins and kinases to the glomerular basement membrane and adapter proteins of slit diaphragm insertion sites. Methods To investigate whether high-glucose (HG) and advanced glycosylation end products (AGEs) induce quantitative and distributional changes of podocyte p130Cas protein, a docking protein connecting F-actin fibers to the glomerular basement membrane and adapter proteins, we cultured rat GECs (r-GECs) and mouse GECs (m-GECs) under (i) normal glucose (5 mM = control), (ii) HG (30 mM), (iii) AGE-added or (iv) HG plus AGE-added conditions. We also prepared streptozotocin-induced diabetic renal tissues. Results We found that p130Cas stainings were located in peripheral cytoplasm of r-GECs and m-GECs as dots in linear alignment, partially colocalized with actin-binding proteins such as synaptopodin and alpha-actinin, and locally connected to the ends of actin filaments. In diabetic conditions, the intensities of p130Cas stainings were diminished in more pathological HG plus AGE-added condition of r-GECs and m-GECs and in chronic diabetic renal tissues. p130Cas protein was decreased significantly by HG, AGE and HG plus AGE in both cells compared with normal glucose or osmotic control conditions. p130Cas mRNA expression levels were also suppressed similarly in diabetic conditions. Conclusion We suggest that diabetic conditions modulate the quantitative and distributional changes of podocyte p130Cas and therefore affect the filtration function of podocytes.

Sigumjang made from barley bran is a traditional fermented food only found in Gyungsang-do area of South Korea. Appearance, tastes and flavors of sigumlang differs from Doenjang (well-known Korean traditional fermented soybean paste). We investigated characteristics of sigumjang and isolated strains of microorganism with high enzymatic activity from sigumjang meju. We collected 19 kinds of sigumjang and 14 kinds of sigumjang meju, which is dried-fermented barley bran and serves as the basis of sigumjang. Methods of sigumjang meju-making are different by region from baking directly the dough of barley bran or after steaming the dough. Sigumjang showed lower pH of 4.37 - 5.99 compared to other Korean traditional fermented jang, because of its lactic acid fermentation. 13 strains showed high hydrolysis activity of cellulose, starch, protein among 51 strains collected from the 14 kinds of sigumjang meju. SM-26 stain showed a high enzymatic activity of cellulase, amylase and protease. SM-26 stain was identified as Bacillus amyloliquefaciens subsp. based on its 16S rRNA sequence analysis.

Proteinuria is typically accompanied by structural and compositional changes of the foot processes and of the slit diaphragms between podocytes. CD2-associated protein (CD2AP) in podocytes serves as an adaptor protein binding to nephrin and podocin, anchoring these slit diaphragm proteins to actin filaments of podocyte cytoskeleton and sending signals inward or outward.In the present study, we prepared streptozotocin-induced diabetic renal tissues and cultured podocytes in diabetic conditions to investigate podocyte phenotypical changes, including quantitative and distributional changes of CD2AP protein and search for the signalling mechanisms in diabetic conditions. We prepared cultured rat glomerular epithelial cells and mouse podocytes to study how high glucose and advanced glycosylation end products (AGE) induce phenotypical changes of cultured podocyte, under (1) normal glucose (5 mM, = control), (2) high glucose (30 mM), (3) AGE-added or (4) high glucose plus AGE-added conditions.According to diabetic duration, density of CD2AP in renal tissue of experimental diabetic nephropathy became conglomerulated and diminished. In cultured podocytes, CD2AP co-localized with nephrin and zonula occludens-1 by confocal imaging. High glucose and high glucose plus AGE induced the relocalization and concentration of CD2AP at internal cytoplasmic and perinuclear areas of podocytes. High glucose plus AGE-added condition also decreased CD2AP protein amount and its mRNA expression compared with normal glucose or osmotic control conditions. In addition, LY294002, a phosphoinositide 3-kinase inhibitor, prevented the quantitative and distributional changes of CD2AP induced by high glucose and AGE.These findings suggest that diabetic conditions induce the phenotypical changes of podocyte CD2AP possibly via phosphoinositide 3-kinase/Akt signalling.

In this study, we investigated whether fermented rice bran (FRB) can ameliorate the oxidative stress induced by high glucose and hydrogen peroxide (H(2)O(2)) in 3T3-L1 adipocytes by analyzing reactive oxygen species (ROS), oil red O staining, as well as the expression of mRNAs related to glucose homeostasis and adipogenesis. It was first confirmed that rice bran fermented by Issatchenkia orientalis MFST1 extract increased free phenolic content compared to non-fermented rice bran. The FRB extract strongly inhibited ROS generation and upregulated the expression of PPAR-γ and adiponectin. Moreover, FRB upregulated GLUT4 related to glucose transportation and insulin sensitivity. Taken together, FRB extract ameliorated oxidative stress-induced insulin resistance by neutralizing free radicals and upregulating adiponectin in adipocytes. Our results provide information toward understanding the beneficial effects of FRB on oxidative stress.

The slit diaphragm connecting the adjacent foot processes of glomerular epithelial cells (podocytes) is the final barrier of the glomerular capillary wall and serves to prevent proteinuria. Podocytes are understood to be terminally differentiated cells and share some common features with neurons. Neurexin is a presynaptic adhesion molecule that plays a role in synaptic differentiation. Although neurexin has been understood to be specifically expressed in neuronal tissues, we found that neurexin was expressed in several organs. Several forms of splice variants of neurexin-1α were detected in the cerebrum, but only one form of neurexin-1α was detected in glomeruli. Immunohistochemical study showed that neurexin restrictedly expressed in the podocytes in kidneys. Dual-labeling analyses showed that neurexin was colocalized with CD2AP, an intracellular component of the slit diaphragm. Immunoprecipitation assay using glomerular lysate showed that neurexin interacted with CD2AP and CASK. These observations indicated that neurexin localized at the slit diaphragm area. The staining intensity of neurexin in podocytes was clearly lowered, and their staining pattern shifted to a more discontinuous patchy pattern in the disease models showing severe proteinuria. The expression and localization of neurexin in these models altered more clearly and rapidly than that of other slit diaphragm components. We propose that neurexin is available as an early diagnostic marker to detect podocyte injury. Neurexin coincided with nephrin, a key molecule of the slit diaphragm detected in a presumptive podocyte of the developing glomeruli and in the glomeruli for which the slit diaphragm is repairing injury. These observations suggest that neurexin is involved in the formation of the slit diaphragm and the maintenance of its function.

High glucose levels can change podocyte gene expression and subsequently induce podocyte damage through altered glucose metabolism. l-Carnitine is known to play a beneficial role in diabetes; however, there are no studies on the effects of l-carnitine on podocyte alteration under high glucose conditions. This study investigated whether l-carnitine can attenuate diabetic podocyte injury through the prevention of loss of slit diaphragm proteins. The l-carnitine treatment group showed increased glucose uptakes compared to the control group, suggesting that glucose utilization in the podocytes was increased by l-carnitine. l-Carnitine treatment also prevented decreased mRNA expressions of nephrin and podocin in the high glucose-stimulated podocytes. However, mRNA expressions of CD2AP and α-actinin-4 were not significantly changed by the high glucose conditions. When these data are taken together, l-carnitine can increase glucose uptake in podocytes under high glucose conditions, and its mechanism may be at least partly related to the up-regulation of nephrin and podocin. Our results help clarify the beneficial effects of l-carnitine in diabetic nephropathy.

Recent studies have demonstrated that the slit diaphragm of the glomerular epithelial cell (podocyte) is the structure likely to be the barrier in the glomerular capillary wall. Murine monoclonal antibody against nephrin, a molecule constituting the extracellular site of the slit diaphragm, caused severe proteinuria if injected into rats, in a complement- or inflammatory cell-independent manner. In this proteinuric state, not only nephrin but also other slit diaphragm-associated molecules are down-regulated. These observations suggest that the antibody alters the molecular composition of the slit diaphragm and, thereby, affects the glomerular permeability barrier. Recently, it was found that IP-10, SV2B, ephrin B1 and the receptors of angiotensin II were expressed in the podocyte, and that their expressions were clearly altered in anti-nephrin antibody-induced nephropathy. It is conceivable that these molecules are involved in the development of proteinuria in this model. IP-10 is assumed to play a role in maintaining the slit diaphragm function by regulating the cell cycle balance of the podocyte. SV2B and ephrin B1 play pivotal roles in the proper localization of the slit diaphragm component. In vivo and in vitro studies demonstrated that angiotensin II type 2 receptor-mediated action enhanced the expression of nephrin. We propose that these molecules could be novel therapeutic targets for proteinuria.