Immunobiotic lactic acid bacteria beneficially regulate immune response triggered by poly(I:C) in porcine intestinal epithelial cells

ABSTRACT: This study analyzed the functional expression of TLR3 in various gastrointestinal tissues from adult swine and shows that TLR3 is expressed preferentially in intestinal epithelial cells (IEC), CD172a+CD11R1high and CD4+ cells from ileal Peyer's patches. We characterized the inflammatory immune response triggered by TLR3 activation in a clonal porcine intestinal epitheliocyte cell line (PIE cells) and in PIE-immune cell co-cultures, and demonstrated that these systems are valuable tools to study in vitro the immune response triggered by TLR3 on IEC and the interaction between IEC and immune cells. In addition, we selected an immunobiotic lactic acid bacteria strain, Lactobacillus casei MEP221106, able to beneficially regulate the anti-viral immune response triggered by poly(I:C) stimulation in PIE cells. Moreover, we deepened our understanding of the possible mechanisms of immunobiotic action by demonstrating that L. casei MEP221106 modulates the interaction between IEC and immune cells during the generation of a TLR3-mediated immune response.     

Development of winter wheat with excellent resistance to pre-harvest sprouting and rain damage

Pre-harvest sprouting and high alpha-amylase activity are the most serious problems for the wheat production in Hokkaido, the northern most part of Japan. It has become more frequent for wheats to be exposed to continuous rain for several days in harvest time. In addition, low temperature below 15 ^°C, combined with rainfall, accelerates the damage of pre-harvest sprouting. Therefore, we started a breeding program in 1998 to develop wheat varieties with excellent resistant to sprouting damage in Hokkaido.Selection and evaluation of pre-harvest sprouting of resistant lines were conducted by two tests. One test was artificial rain treatment of intact spikes and the other was a germination test of hand-threshed grains at 10, 15 and 20 ^°C. The spikes of each line and variety were collected at physiological maturity and at one week after the first sampling date. Falling number was also measured.We have succeeded in breeding a deeply dormant breeding line, Kitakei 1802, from 1,923 doubled haploid lines raised by anther culture from the cross between Kitakei 1616 and Nishikazekomugi. Kitakei 1802 hardly sprouted through 10 days” continuous rainfall treatment at low temperature (15 ^°C) and maintained high falling number (> 300 s) in 2002 and 2003.     

培养基对尖孢镰刀菌非致病力菌株281产孢量的影响

选择8种培养基，采用液体培养法，研究培养基质对尖孢镰刀菌(Fusarium oxysporum Schl.)281菌株小孢子产生量的影响，结果表明，在PDA培养基上培养144h得到最大小孢子产量1.283×109cfu/ml，与其它培养基产孢量差异显著（P=0.05）。培养至192h时，ATCC培养基的小孢子数量达到1.325×109cfu/ml，与PDA培养基之间无显著差异（P=0.05），与其它6种培养基之间差异显著（P=0.05）。综合比较8种培养基，PDA培养基培养时间短，配制简单，是较为理想的产小孢子培养基。     

Production of saturated acyl L-ascorbate by immobilized lipase using a continuous stirred tank reactor

6-O-decanoyl, 6-O-dodecanoyl, or 6-O-tetradecanoyl L-ascorbate was continuously produced at 50 degrees C using a continuous stirred tank reactor (CSTR) with an immobilized lipase, Chirazyme L-2 C2, from Candida antarctica. Acetone was used as the reaction medium. For each saturated acyl L-ascorbate, the productivity of ca. 60 g/L reactor/day was achieved for at least 11 days. The solubility of the saturated acyl L-ascorbate in the soybean oil or water was measured at various temperatures. The solubilities in both the soybean oil and the water were higher for L-ascorbate with a shorter acyl chain. The acyl chain dependence of the solubility in water was stronger than that of the solubility in soybean oil. The temperature dependences of the solubility in both soybean oil and water could be expressed by the van't Hoff equation, and the dissolution enthalpy (DeltaH) values for the soybean oil and water were about 20 and 90 kJ/mol, respectively, irrespective of the acyl chain length. The radical scavenging activities of L-ascorbic acid and the saturated acyl L-ascorbates against 1,1-diphenyl-2-picrylhydrazyl free radical were ca. 95% for all of the compounds, and the introduction of a saturated acyl group to the L-ascorbic acid did not affect the activity.     

Yield and quality evaluations on a pair of processing tomato lines nearly isogenic for the Tm2a gene for resistance to the tobacco mosaic virus

A pair of processing tomato (Lycopersicon esculentum Mill.) lines, nearly isogenic for the Tm-2^a gene for resistance to tobacco mosaic virus, were grown in replicated trials under commercial production conditions in five locations worldwide. The lines were evaluated for 17 processing traits including fruit yield, size, soluble solids concentration, color, firmness, and viscosity. Eight of those traits differed significantly among the nearly-isogenic lines (NILs). Most notably, the NIL heterozygous for Tm-2^a yielded, on average, 16% more than the NIL homozygous for the susceptible allele and 33% more than the NIL homozygous for Tm-2^a. Viscosity was lower in the heterozygous NIL and the homozygous and heterozygous resistant NILs had softer fruit with larger stem scars compared to the homozygous susceptible NIL. These results indicate the presence of the Tm-2^a locus affects many traits of importance for processing tomatoes and may be used, in the heterozygous state, to significantly increase yield. Whether the observed effects are due to the Tm-2^a gene itself or genes associated via linkage drag could not be determined. This revised version was published online in July 2006 with corrections to the Cover Date.     

Stopped-flow kinetic study of the aroxyl radical-scavenging action of catechins and vitamin C in ethanol and micellar solutions

Kinetic study of the aroxyl radical-scavenging action of catechins (epicatechin (EC), epicatechin gallate (ECG), epigallocatechin (EGC), and epigallocatechin gallate (EGCG)) and related compounds (methyl gallate (MG), 4-methylcatechol (MC), and 5-methoxyresorcinol (MR)) has been performed. The second-order rate constant ( k s) for the reaction of these antioxidants with aroxyl radical has been measured in ethanol and aqueous Triton X-100 micellar solution (5.0 wt %). The k s values decreased in the order of EGCG > EGC > MC > ECG > EC > MG > MR in ethanol, indicating that the reactivity of the OH groups in catechins decreased in the order of pyrogallol B-ring > catechol B-ring > gallate G-ring > resorcinol A-ring. The structure-activity relationship in the free radical-scavenging reaction by catechins has been clarified by the detailed analyses of the pH dependence of k s values. From the results, the p K a values have been determined for catechins. The monoanion form at catechol B- and resorcinol A-rings and dianion form at pyrogallol B- and gallate G-rings show the highest activity for free radical scavenging. It was found that the free radical-scavenging activities of catechins are 3.2-13 times larger than that of vitamin C at pH 7.0.     

Therapeutic potential of canine bone marrow stromal cells (BMSCs) in the carbon tetrachloride (CCl4) induced chronic liver dysfunction mouse model

Regenerative medicine using bone marrow cells is an attractive therapy for the cure of patients with severe liver disease. Here, we show the therapeutic potential of canine bone marrow stromal cells (BMSCs) in mouse models of CCl(4)-induced chronic liver dysfunction. We used two different models for xenotransplantation, nude mice and cyclosporine A (CSA) immunosuppressed mice. Serum parameters from a standard liver panel were not improved following transplantation. However, fibrotic liver lesions with severe inflammation were decreased in CCl(4)-treated CSA mice following BMSC transplantation. Effective migration of transplanted canine BMSCs was limited to persistently injured liver in CCl(4)-treated CSA mice, where they may be effective in resolving inflammatory fibrotic lesions. These results suggest that canine BMSCs are an effective cell source for liver regeneration.     

Structural differences between myofibrillar protein,paratropomyosin, and tropomyosin as revealed by high‐performance liquid chromatography

Paratropomyosin (PTM) composes myofibril functions to weaken the rigor linkages formed between actin and myosin during postmortem aging of muscles. PTM has the similar physico‐chemical properties as tropomyosin (TM) that is a regulatory protein of myofibrils. So far, it is unclear whether PTM is definitely different from TM, because the primary structure of PTM has not been determined yet. The aim of this study was to clarify structural difference of PTM from TM. PTM was prepared by column chromatography immediately after slaughter from broiler breast muscle, and purified by high‐performance liquid chromatography (HPLC). Purified PTM was successfully separated from TM, and the recovered PTM molecule was reduced with dithiothreitol to separate again by HPLC. Two subunits were obtained and peptides from each digested subunit by V8 protease were recovered by HPLC, and then amino acid sequences of the peptides were analyzed by protein sequencing. As a result, some amino acid residues were replaced from that of TMα1 isoform which is the major isoform of TM, and also was different between the two subunits. Therefore, it is concluded that PTM clearly differs from TM and it is suggested that functional difference in PTM from TM is attributed to amino acid replacements in subunits composing PTM.     

Differential expression of neurofilament 200-like immunoreactivity in the main olfactory and vomeronasal systems of the Japanese newt, Cynops pyrrhogaster

Expression of neurofilament 200 (NF200)-like immunoreactivity was examined in the main olfactory system and the vomeronasal system of the Japanese newt, Cynops pyrrhogaster, using anti-porcine NF200 monoclonal antibody (clone N52) to investigate the differences in phenotypical characteristics between these systems. The entire nasal cavity was a flattened single chamber consisting of the main nasal chamber (MNC) and the lateral nasal sinus (LNS) communicating with each other. The olfactory epithelium (OE) was present in the MNC, and the vomeronasal epithelium (VNE) was in the LNS. The OE possessed only a small number of NF200-like immunoreactive receptor neurons. The olfactory nerve and the olfactory nerve layer of the main olfactory bulb also contained a small number of NF200-like immunoreactive axons. In contrast, the VNE possessed many NF200-like immunoreactive receptor neurons. The vomeronasal nerve and the vomeronasal nerve layer of the accessory olfactory bulb contained many NF200-like immunoreactive axons. These findings in the Japanese newt indicate that NF200-like immunoreactive receptor neurons constitute a major subpopulation in the VNE and a minor subpopulation in the OE. In addition, NF200-like immunoreactivity seems to be a useful marker to distinguish the vomeronasal system from the other nervous systems including the main olfactory system in the Japanese newt. The localization of a few NF200-like immunoreactive receptor neurons in the OE might indicate that pheromone-sensitive receptor neurons are intermingled in the OE of the Japanese newt.