Molecular cloning and characterization of chicken tumor necrosis factor (TNF)-superfamily ligands, CD30L and TNF-related apoptosis inducing ligand (TRAIL)

CD30 ligand (CD30L) and tumor necrosis factor (TNF)-related apoptosis inducing ligand (TRAIL) are members of the TNF-superfamily that have many important biological activities in cell proliferation and apoptotic death. In this study, both genes in the chicken were cloned and their expression was analyzed. Complementary DNA fragments were obtained from a suppressive subtractive hybridization library with or without lipopolysaccharide (LPS)-stimulation. Chicken CD30L consists of 1,152 base pairs (bp) with an open reading frame (ORF) of 720 bp having 36.4% identity with human CD30L, whereas chicken TRAIL is 1,134 bp long with an ORF of 912 bp having 54.4% identity with human TRAIL. Chicken CD30L was expressed at high levels in the spleen, bursa of Fabricius and in the chicken monocytic leukemia cell line, IN24. Stimulation with LPS in the spleen, bursa of Fabricius and the IN24 cell line did not affect CD30L expression. The gene expression of chicken TRAIL was essentially to the same level in all tissues examined. The time course of expression was not significantly altered by LPS-stimulation in the spleen, thymus and bursa of Fabricius, but reached a maximal level 8 hr after stimulation in the IN24 cell line. The high level expression of both genes in lymphoid organs and IN24 cell line indicates that chicken CD30L and TRAIL may also play an important role in apoptotic signal transduction and the regulation of cell proliferation in the immune system.     

Evaluation of equine cecal motility by ausculation, ultrasonography and electrointestinography after jejunocecostomy

OBJECTIVE: Horses often suffer reduced intestinal motility after jejunocecostomy. Therefore, accurate evaluation of intestinal motility is important for the prevention, diagnosis and treatment of this condition. The purpose of this study was to evaluate intestinal motility in horses after jejunocecostomy using three different methods, i.e. auscultation, ultrasonography and electrointestinography. ANIMALS: Six healthy thoroughbreds were used in this study. They were subjected to jejunocecostomy. PROCEDURE: Bowel sounds in the right paralumbar fossa were assigned a score of 0-3 for intestinal motility evaluation by auscultation, and the number of cecal contractions during a 3-min period were counted by ultrasonography. Electrointestinography (EIG) was used to measure percutaneous potential of the cecum. RESULTS: We identified three specific postoperative periods: the period of reduced intestinal motility (postoperative day 1 to day 2), in which intestinal motility declined, the unstable period (day 3 to day 7), in which intestinal motility partially recovered, and the full recovery period (day 8 to day 31), in which intestinal motility returned to preoperative state. Careful management was found to be especially important during the period of reduced intestinal motility and the unstable period. We found that, in healthy horses that underwent jejunocecostomy, it takes approximately one month for the cecum to return to normal motility patterns observed before surgery. CONCLUSION: We have shown in this study that evaluation of intestinal motility after jejunocecostomy in horses by EIG is more objective and provides more details than evaluation by auscultation or ultrasonography.     

Metabolism of N-[(R)-1-(2,4-dichlorophenyl)ethyl]-2-cyano-3,3-dimethylbutanamide (Delaus, S-2900) and its isomer, N-[(S)-1-(2,4-dichlorophenyl)ethyl]-2-cyano-3,3-dimethylbutanamide (S-2900S), in rats. 1. Identification of metabolites in feces and urine

Rats were orally dosed with a 1:1 diastereomixture of N-[(R)-1-(2,4-dichlorophenyl)ethyl]-2-cyano-3,3-dimethylbutanamide (Delaus, S-2900) and N-[(S)-1-(2,4-dichlorophenyl)ethyl]-2-cyano-3,3-dimethylbutanamide (S-2900S), both labeled with 14C, at 200 mg/kg/day for 5 consecutive days, and 16 metabolites in urine and feces were purified by a combination of several chromatographic techniques. The chemical structures of all isolated metabolites were identified by spectroanalyses (NMR and MS). Several of them were unique decyanated and/or cyclic compounds (lactone, imide, cyclic amide, cyclic imino ether forms). Major biotransformation reactions of the mixture of S-2900 and S-2900S in rats are proposed on the basis of the metabolites identified in this study.     

Effect of Dietary Inclusion of Blue Mussel Extract on Growth and Body Composition of Japanese Flounder Paralichthys olivaceus

Dietary inclusion of a water-soluble fraction of blue mussel Mytilus galloprovincialis was examined as a feeding stimulant for juvenile Japanese flounder Paralichthys olivaceus . The control diet mainly consisted of fish meal, potato starch, and pollack liver oil. Five, 10, and 20% (weight/weight) of the control diet was exchanged with aqueous extracts of blue mussel meat in experimental groups. Fish of about 10 g in initial body weight were fed each diet to satiation, twice daily, 6 d per wk for 6 wk at 20 C. The final body weight, weight gain, and feed efficiency of fish fed the diets containing blue mussel extracts were significantly higher than those of fish fed the control diet. However, these parameters were not different among experimental groups containing blue mussel extract independent of the inclusion level of extract. A similar trend was shown in protein efficiency ratio as fish fed the control diet had a significantly lower protein efficiency ratio than the other dietary groups. Compared to the control diet, higher plasma protein and lower triglyceride were found in fish fed the diets with the extract, while other blood constituents were relatively similar for the dietary groups tested. On the other hand, whole-body crude lipid content and lipid retention of fish fed the diets with the extract were generally significantly higher than those of fish fed the control diet. Whole body crude protein was identical regardless of the dietary composition; however, protein retention of fish showed a similar trend to lipid retention.     

Lysine decarboxylase-negative Salmonella enterica serovar enteritidis: antibiotic susceptibility, phage and PFGE typing

One hundred twenty Salmonella Enteritidis isolates collected from 1992 to 2005 in Nagasaki prefecture (65 isolates from 40 outbreak cases, 44 from sporadic diarrhea patients, and 11 from chicken-related products) were investigated by their antibiotic susceptibility profiles, phage typing, and pulsed-field gel electrophoresis (PFGE) typing. Out of them, 18 were identified as lysine decarboxylase (LDC)-negative isolates, and 15 showed resistance toward streptomycin. Based on the PFGE typing, the isolates were classified into five clusters by UPGMA clustering method. Three LDC-negative isolates belonged to cluster A and were of phage type (PT) 4 and isolated between 2000 and 2004. Other 15 LDC-negative isolates belonged to cluster E. They were PT1, reacted but did not conform (RDNC), or untypable and were isolated between 2001 and 2004. LDC-negative isolates of the cluster A differed from LDC-negative isolates of the cluster E in antibiotic susceptibility profiles, phage typing, and PFGE typing. LDC-negative isolates of the cluster E were isolated after 2001 in Nagasaki prefecture.     

Metabolism of 7-fluoro-6-(3,4,5,6-tetrahydrophthalimido)-4- (2-propynyl)-2H-1,4-benzoxazin-3(4H)-one (S-53482, flumioxazin) in the rat: II. Identification of reduced metabolites

On single oral administration of (14)C-S-53482 [7-fluoro-6-(3,4,5, 6-tetrahydrophthalimido)-4-(2-propynyl)-2H-1,4-benzoxazin-3( 4H)-one, Flumioxazin] labeled at the 1- and 2-positions of tetrahydrophthaloyl group to rats at 1 (low dose) or 100 (high dose) mg/kg, the radiocarbon was almost completely eliminated within 7 days after administration in both groups with generally very low residual (14)C tissue levels. The predominant excretion route was via the feces. The major fecal and urinary metabolites involved reduction or sulfonic acid addition reactions at the 1,2-double bond of the 3,4,5,6-tetrahydrophthalimide moiety and hydroxylation of the cyclohexene or cyclohexane ring. One urinary and four fecal metabolites were identified using chromatographic techniques and spectroanalyses (NMR and MS). Three of five identified metabolites were unique forms, reduced at the 1,2-double bond of the 3,4,5, 6-tetrahydrophthalimide moiety. On the basis of the metabolites identified in this study, the metabolic pathways of S-53482 in rats are proposed. To specify tissues forming reduced metabolites, an in vitro study was conducted. Reduction was found to take place in red blood cells.     

Behavior of phenolic substances in the decaying process of plants

The changes in amount of ether-extractable phenolic substances, individual phenolic adds, organic matter content, and pH value in the decaying process of rice straw were compared at 10, 30, and 50°C under moist and flooded conditions for 70 days.

1) The changes in pH value with time varied markedly according to temperature and water conditions.

2) Phenolic substances in the ether-extractable fraction obtained from the acidified solution of alkaline methanol extract contained undetected phenolic compounds in amounts larger than the phenolic adds detected by gas chromatography. pH value during incubation was hardly attributable to these phenolic substances because of their small quantity.

3) The changes in amount of the ether-extractable phenolic substances with time was mainly due to phenolic compounds other than phenolic acids, and influenced largely by the temperature and water conditions. Their formation and degradation took place rapidly at high temperatures under flooded conditions.

4) From the plots of individual phenolic acids vs. time, it was tentatively concluded that phenolic acids contalned in non-decayed straw were rapidly degraded in the earlier stage, and some of them were newly produced from ether-unextractable precursors in the later stage. These reactions occurred rapidly at high temperatures, especially under flooded conditions.

5) The content of p-hydroxybenzolc acid increased simultaneously with a rapid decrease in p-coumaric acid during the incubation ot straw. The relationship between vanillic acid and ferulic acid was the same.     

Eliminating the effect of epidural fat during dorsolumbar epidural analgesia in cattle

Objective To compare the distribution of new methylene blue (NMB) solution injected into either the subperiosteal space or under the epidural fat at the first and second lumbar vertebral interspace in cattle. Study design Prospective experimental study. Sample population Nine nonpregnant cows. Methods Needles were advanced towards the first interlumbar epidural space using a dorsal mid‐line approach. Upon confirmation of successful penetration of the ligamentum flavum, group 1 received 5 mL of 0.12% NMB in 0.9% saline. In group 2, the needle was slowly advanced a further 7–10 mm to ensure injection under the epidural fat; the same dose was injected. The extent of cranial and caudal migration of dye, as indicated by staining of epidural fat and the dura mater was measured. Results In group 1 (n = 4), NMB solution migrated only between the periosteum and the epidural fat. There was unilateral distribution in two cows. In contrast, there was no difference between left‐ and right‐sided distribution in group 2 (n = 5), and the NMB solution migrated only between epidural fat and the dura mater. Although the dura mater was not penetrated in any animal of group 2, the dorsal roots were slightly stained and the overlying dura mater was heavily stained in these animals. Conclusions and clinical relevance Variation in distribution of local anaesthetic solution can probably be overcome by deeper needle penetration of the epidural fat.     

Chimeric inheritance of organelle DNA in variegated leaf seedlings from inter-subgeneric crossing of azalea

The inheritance of organelle DNA was investigated using PCR–RFLP markers in reciprocal cross combinations of inter-subgeneric azalea hybrids between evergreen azaleas (Rhododendron nakaharai and its hybrids) and fragrant deciduous azaleas (R. arborescens and R. viscosum) for the purpose of breeding fragrant evergreen azaleas. The hybrid progenies included green leaf, pale green leaf, variegated leaf and albino seedlings. Most viable green leaf seedlings had inherited ptDNA from the deciduous parent and non-viable albino and pale green leaf seedlings had inherited ptDNA from the evergreen parent. On the other hand, variegated leaf seedlings had chimerically inherited ptDNA from both parents. Their green leaf segments had ptDNA from the deciduous parent, and the pale green and white segments had biparental or maternal ptDNA depending on the progeny. In this study, we obtained interesting inter-subgeneric azalea hybrid progenies that had chimerically inherited organelle DNA and had different colored leaf segments corresponding to the composition of ptDNA from each parent. These results suggest that variegated leaf progenies with chimeric ptDNA from both parents can be subsistent, whereas albino seedlings resulting from plastome–genome incompatibility between the plastid genome from evergreen azalea and the nuclear genome from deciduous azalea are non-viable.