• Proceedings of the Korean Society of Applied Pharmacology
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• 1996.11a
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• pp.65-73
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• 1996

LPS/LOS, the compound found only in gram-negative bacterial outer membrane, plays important roles in bacterial maintenance as well as its pathogenesis. We isolated and characterized several genes required for NTHi 2019 LOS biosynthesis, which encode enzymes required for sugar substrate synthesis or the transfer of substrates to receptor molecules. The htrB gene, however, appears to have more complex role. It has acryltransferase activity as well as various other activity, which may control regulation of LOS biosynthesis as well as its pathogenicity. Evidences supporting the latter come from the observations that the lipid A of the B29 induced significantly less TNF ${\alpha}$ from macrophages than that of the wild type LOS (unpublished data). H. influenzae A2-htrB mutant strain was also significantly less invasive than the wild type strain. The structural similarities of the enterobacterial LPS and the Haemophilus LOS enabled us to isolate the NTHi 2019 genes involved in LOS biosynthesis genes by using the S. typhimurium LPS deep core mutants. While a similar approach has been used for E. coli, this technique for selection of an LPS phenotype has not been applied to nonenterobacterial species. The difficulties inherent in the molecular manipulation of organism such as Neisseria and Haemophilus species make this approach particularly attractive in the identification and cloning LOS genes. Studies on genetic features of LPS/LOS biosynthesis would be useful for understanding bacterial pathogenesis as well as for developing vaccines for these gram-negative pathogenic bacteria.

• Korean Journal of Veterinary Service
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• v.33 no.3
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• pp.223-231
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• 2010

A total of 191 samples collected from the commercial swine farms located in Chungnam province were investigated by PCR to estimate the prevalence of Lawsonia (L.) intracellularis infection. In the group of the pigs with proliferative enteritis, 14 (93.3%) of 15 intestinal samples and 12 (80.0%) of 15 feces were positive in PCR. In contrast, a relatively low positive rate (18.0%, 29 of 161 samples) was determined in the group of normal healthy pigs. The group of pigs over 120 days showed the highest positive rates (26.8%, 15 of 56 samples). In the comparison of the sequences of 210bp for species specific fragments and 301bp for outer membrane protein, the isolates (L1. L2) showed almost 100% identity with the reference L. intracellularis (L08049, USA). For the sequences of partial 16s rDNA, the homologies among the 5 isolates (L1-L5) were 97.4% to 99.3%, and those of 5 sequences (L1-L5) versus 5 overseas reference strains of L. intracellularis ranged from 98.6% to 99.8%. In the comparison of the nucleotide sequences among 5 isolates and other species in Desulfovibrionales showed 82.4 to 99.5% identities. The 5 isolates shared relatively low identities (76.9% to 84.4%) with the species of alpha-proteobacteria. In phylogenetic analysis based on the 16s rDNA sequences, all of the 5 isolates (L1-L5) were located in the same branch with the strains of L. intracellularis that were previously isolated from the pigs in USA and China. Seven strains of Desulfovibrio sp. were clustered in the neighboring branches, whereas alpha and gamma Proteobacteria showed distant relationship with L. intracellularis strains. The present findings suggest that L. intracellularis infection is endemic in the swine farms in the regions, and that the domestic isolates maintained very limited genetic variation.

• Molecules and Cells
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• v.38 no.5
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• pp.402-408
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• 2015

Protein O-GlcNAcylation, dictated by cellular UDP-N-acetylglucosamine (UDP-GlcNAc) levels, plays a crucial role in posttranslational modifications. The enzyme GlcNAc kinase (NAGK, E.C. 2.7.1.59) catalyzes the formation of GlcNAc-6-phosphate, which is a major substrate for the biosynthesis of UDP-GlcNAc. Recent studies have revealed the expression of NAGK in different types of cells especially in neuronal dendrites. Here, by immunocytochemistry (ICC) and immunonucleochemistry (INC) of cultured rat hippocampal neurons, HEK293T and GT1-7 cells, we have showed that NAGK immuno-reactive punctae being present in the nucleoplasm colocalized with small nuclear ribonucleoprotein-associated protein N (snRNPN) and p54NRB, which are speckle and paraspeckle markers, respectively. Furthermore, NAGK IR cluster was also found to be colocalized with GTF2H5 (general transcription factor IIH, polypeptide 5) immuno reactive punctae. In addition, relative localization to the ring of nuclear lamin matrix and to GlcNAc, which is highly enriched in nuclear pore complexes, showed that NAGK surrounds the nucleus at the cytoplasmic face of the nuclear outer membrane. By in situ proximity ligation assay (PLA) we confirmed the colocalization of NAGK with snRNPN in the nucleus and in dendrites, while we also verified the interactions of NAGK with p54NRB, and with GTF2H5 in the nucleus. These associations between NAGK with speckle, paraspeckle and general transcription factor suggest its regulatory roles in gene expression.

• Korean Journal of Veterinary Research
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• v.45 no.4
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• pp.593-599
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• 2005

An abattoir study on the abdominal fat necrosis in adult cattle was performed pathologically. Grossly, masses of fat necrosis were leekgreen in colour, lobulated on the cut surface, and saponificated in the texture. These necrotic adipose tissues infiltrated usually into neighboring parenchymal organs including intestines and pancreas, leading to fibrosis or atrophy of them. Histopathologically, necrotic fat cells contained acidophilic, opaque, amorphous substance or basophilic fibrillar or granular minerals in their cytoplasms. The lesions of fat necrosis were divided by fibroconnective tissue. With increase of the severity, necrotic fat cells fused each other and then formed fat cysts. In this severe lesion, necrotic fat cells were partialy or completely replaced by macrophages. Multinucleated giant cells were scattered in this lesion. Interestingly, small artery in the lesion of fat necrosis revealed severe thickening of internal elastic membrane. Severe fibrosis was observed in or between the outer longitudinal and inner circular muscular externas causing segregation, degeneration and necrosis of muscle fibers. The nerve cells of Auerbach's and Meissner's plexuses surrounded by fibrosis were degenerated or necrotic. In addition, necrotic fat cells infiltrated into the pancreas, resulting in pancreas atrophy. From these results, it is speculated that fat necrosis might compromise intestinal movement due to necrosis of muscular externa and ganglion cells of Auerbach's and Meissner's plexuses.

• Korean Journal of Veterinary Research
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• v.33 no.3
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• pp.465-469
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• 1993

This paper dealt with the histopathological findings on the natural cysticercosis in pigs. Three cases of porcine cysticercosis, which had been kept in the Department of Veterinary Medicine, Cheju. University more than ten years, were histopathologically examined in order to see the host reaction to the parasite. Capsules containing scolex were mainly found in the fascia of skeletal muscle, heart, and brains. Microscopically, cysticerci in the epicardium and the fascia of skeletal muscles were encapsulated with fibroblasts and collagen fibers. Around capsules, there was infiltration of eosinophils, lymphocytes and macrophages, although the degree and severity of inflammatory reaction varied case by case. Cerebral cortex also had the inflammatory exudate of lymphoid cells in the vicinity of the scolex. whereas perivascular lymphocytic cuffings were commonly seen around capsules. GFAP immunoreactive fibers formed a limiting membrane along the outer side of capsules. There was also proliferation of GFAP-positive astrocytes encirling infiltrating lymphocytes around vessels. In the central nervous system, astrocytes and lympoid cells play an important role in the demarcation of cysts and local immunity, respectively. In conclusion, host tissue reaction in porcine cysticercosis seemed to vary significantly according to the affected organs of pigs. It is assumed that capsules containing worms seemed to be formed at early stage of cysticercosis.

• Journal of Applied Biological Chemistry
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• v.58 no.3
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• pp.227-232
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• 2015

A microarray study has been employed to understand changes of gene expression in E. coli KD43162 resistant to ampicillin, ampicillin-sulbactam, piperacillin, piperacillin-tazobactam, cefazolin, cefepime, aztreonam, imipenem, meropenem, gentamicin, tobramycin, ciprofloxacin, levofloxacin, moxifloxacin, fosfomycin, and trimethoprim-sulfamethoxazole except for amikacin using disk diffusion assay. Using Sodium dodecyl sulphate-polyacrylamide gel electrophoresis and MALDI-TOF MS analyses, 36 kDa of outer membrane proteins (OMPs) was found to be deleted in the multidrug resistant E. coli KD 43162. Microarray analysis was used to determine up- and down-regulated genes in relation to multidrug resistant E. coli KD43162. Among the up-regulated genes, these genes were corresponded to express the proteins as penicillin-binding proteins (PBPs), tartronate semialdehyde reductase, ethanolamine utilization protein, shikimate kinase I, allantoinase, predicted SAM-dependent methyltransferase, L-glutamine: D-fructose-6-phosphate aminotransferase (GFAT), phospho-glucosamine mutase, predicted N-acetylmannosamine kinase, and predicted N-acetylmannosamine-6-P epimerase. Up-regulation of PBPs, one of primary target sites of antibiotics, might be responsible for the multidrug resistance in E. coli with increasing amount of target sites. Up-regulation of GFAT enzyme may be related to the up-regulation of PBPs because GFAT produces N-acetylglucosamine, a precursor of peptidoglycans. One of GFAT inhibitors, azaserine, showed a potent inhibition on the growth of E. coli KD43162. In conclusion, up-regulation of PBPs and GFATs with the loss of 36 kDa OMP refers the multidrug resistance in E. coli KD 43162.

• The Korean Journal of Pain
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• v.28 no.1
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• pp.4-10
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• 2015

Etifoxine (etafenoxine, $Stresam^{(R)}$) is a non-benzodiazepine anxiolytic with an anticonvulsant effect. It was developed in the 1960s for anxiety disorders and is currently being studied for its ability to promote peripheral nerve healing and to treat chemotherapy-induced pain. In addition to being mediated by $GABA_A{\alpha}2$ receptors like benzodiazepines, etifoxine appears to produce anxiolytic effects directly by binding to ${\beta}2$ or ${\beta}3$ subunits of the $GABA_A$ receptor complex. It also modulates $GABA_A$ receptors indirectly via stimulation of neurosteroid production after etifoxine binds to the 18 kDa translocator protein (TSPO) of the outer mitochondrial membrane in the central and peripheral nervous systems, previously known as the peripheral benzodiazepine receptor (PBR). Therefore, the effects of etifoxine are not completely reversed by the benzodiazepine antagonist flumazenil. Etifoxine is used for various emotional and bodily reactions followed by anxiety. It is contraindicated in situations such as shock, severely impaired liver or kidney function, and severe respiratory failure. The average dosage is 150 mg per day for no more than 12 weeks. The most common adverse effect is drowsiness at the initial stage. It does not usually cause any withdrawal syndromes. In conclusion, etifoxine shows less adverse effects of anterograde amnesia, sedation, impaired psychomotor performance, and withdrawal syndromes than those of benzodiazepines. It potentiates $GABA_A$ receptor-function by a direct allosteric effect and by an indirect mechanism involving the activation of TSPO. It seems promising that non-benzodiazepine anxiolytics including etifoxine will replenish shortcomings of benzodiazepines and selective serotonin reuptake inhibitors according to animated studies related to TSPO.

• Journal of fish pathology
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• v.21 no.3
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• pp.201-208
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• 2008

We compared the pathogenicity and antigenicity of two different Edwardsiella tarda (E. tarda) strains KFE and Edk-2 isolated in Korea and Japan respectively. In the pathogenicity analysis with challenge test against loach, E. tarda KFE isolate showed stronger pathogenicity compared to that of E. tarda Edk-2 isolate. The differences were also confirmed by the comparison of OMP (outer membrane protein) in SDSPAGE which showed three major bands, 41kDa, 37kDa and 30kDa, for E. tarda KFE isolates and two major bands, 41kDa and 30kDa, for E. tarda Edk-2 isolates. On the base of these results, we tried to determine the differences of antigenicities of these two isolates in loach which is one of the important species in freshwater aquaculture in Korea. Numbers of specific antibody secreting cells (SASC), appeared to be higher in loach immunized with FKC of E. tarda Edk-2 than loach immunized with FKC of E. tarda KFE. ELISPOT-assay for the comparison of antigenicity showed relatively high percentage of cross-reaction and implied the presence of some common epitopes in the antigens of these two E. tarda isolates.

• The Korean Journal of Malacology
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• v.5 no.1
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• pp.1-9
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• 1989

A scanning electron microscopic study on the glochidium and glchidial encystment of Anodonta grandis on the guppy was conducted. The shape of the glochidium is apparently triangular and its averge size is 0.45mm X0.4mm when closed, The two glochidial shell valves are of the same size, kept together by a ligament of 120${\mu}{\textrm}{m}$ in length and 7 ${\mu}{\textrm}{m}$ in width. Each of the glochidial shell valves has a 16 ${\mu}{\textrm}{m}$ long hook sitdded with many spines on the superior face. A large area to the apex of the valve surrounding the base of the hook is provided with numerous small spines which become progressively smaller towards the periphery of the area, The external surface of the glochidial shell valve is covered with numerous small processes showing successive change in the shape and the pattern of destribution by part. Besides the processes, there are a number of niches scattered all over the exterior surface. The glochidial shell valve has two layers. One is the outer thin membrane bearing the processes and the niches and the others is the inner layer bearing numerous holes which any accessory structure and 2.65 ${\mu}{\textrm}{m}$ in diameter, emerges from a canal located at center of ventral plate of the mamtle, A total of three types of the hair cells are observed. In present artificial infection of the glochidium to the guppy, it took about three to four hours to complete an early cysts, During the period of encystment, The epithelial cells of the host fish actively migrated toward the attached glochidium and covered it.

• Restorative Dentistry and Endodontics
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• v.40 no.4
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• pp.306-313
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• 2015

Objectives: Molecular mechanism of the pathogenicity of Enterococcus faecalis (E. faecalis), a suspected endodontic pathogen, has not yet been adequately elucidated due to limited information on its virulence factors. Here we report the identification of in vivo expressed antigens of E. faecalis by using a novel immunoscreening technique called change-mediated antigen technology (CMAT) and an experimental animal model of endodontic infection. Materials and Methods: Among 4,500 E. coli recombinant clones screened, 19 positive clones reacted reproducibly with hyperimmune sera obtained from rabbits immunized with E. faecalis cells isolated from an experimental endodontic infection. DNA sequences from 16 of these in vivo-induced (IVI) genes were determined. Results: Identified protein antigens of E. faecalis included enzymes involved in housekeeping functions, copper resistance protein, putative outer membrane proteins, and proteins of unknown function. Conclusions: In vivo expressed antigens of E. faecalis could be identified by using a novel immune-screening technique CMAT and an experimental animal model of endodontic infection. Detailed analysis of these IVI genes will lead to a better understanding of the molecular mechanisms involved in the endodontic infection of E. faecalis.