To gain information on retrotransposons in the genome of Paragonimus westermani, PCR was carried out with degenerate primers, specific to protease and reverse transcriptase (rt) genes of long-terminal-repeat (LTR) retrotransposons. The PCR products were cloned and sequenced, after which 12 different retrotransposon-related sequences were isolated from the trematode genome. These showed various degrees of identity to the polyprotein of divergent retrotransposon families. A phylogenetic analysis demonstrated that these sequences could be classified into three different families of LTR retrotransposons, namely, Xena, Bel, and Gypsy families. Of these, two mRNA transcripts were detected by reverse transcriptase-PCR, showing that these two elements preserved their mobile activities. The genomic distributions of these two sequences were found to be highly repetitive. These results suggest that there are diverse retrotransposons including the ancient Xena family in the genome of P. westermani, which may have been involved in the evolution of the host genome.
Amino Acid Sequence ; Animals ; Cloning, Molecular ; DNA, Helminth/analysis ; *Evolution, Molecular ; *Genome ; Molecular Sequence Data ; Paragonimus/*genetics ; Phylogeny ; RNA-Directed DNA Polymerase/chemistry/genetics ; Retroelements/*genetics ; Sequence Alignment ; Sequence Analysis, DNA ; Terminal Repeat Sequences/*genetics

The evolutionary course of the CsRn1 long-terminal-repeat (LTR) retrotransposon was predicted by conducting a phylogenetic analysis with its paralog LTR sequences. Based on the clustering patterns in the phylogenetic tree, multiple CsRn1 copies could be grouped into four subsets, which were shown to have different integration times. Their differential sequence divergences and heterogeneous integration patterns strongly suggested that these subsets appeared sequentially in the genome of C. sinensis. Members of recently expanding subset showed the lowest level of divergence in their LTR and reverse transcriptase gene sequences. They were also shown to be highly polymorphic among individual genomes of the trematode. The CsRn1 element exhibited a preference for repetitive, agenic chromosomal regions in terms of selecting integration targets. Our results suggested that CsRn1 might induce a considerable degree of intergenomic variation and, thereby, have influenced the evolution of the C. sinensis genome.
Animals ; Clonorchis sinensis/*genetics ; DNA, Helminth/analysis/genetics ; *Evolution, Molecular ; Gene Dosage ; *Genome ; Phylogeny ; Polymorphism, Genetic ; RNA-Directed DNA Polymerase ; Retroelements/*genetics ; Sequence Analysis, DNA ; Terminal Repeat Sequences/*genetics

A highly specific antigenic protein of 31 kDa from plerocercoid of Spirometra mansoni (sparganum) was obtained by gelatin affinity and Mono Q anion-exchange column chromatography. The purified 31 kDa protein was subjected to N-glycan enzymatic digestion for structural analysis. The relative electrophoretic mobility was analyzed by SDS-PAGE, before and after digestion. On SDS-PAGE after enzymatic digestion, the 31 kDa protein showed a molecular shift of approximately 2 kDa, which indicated the possession of complex N-linked oligosaccharides (N-glycosidase F sensitive) but not of high-mannose oligosaccharides (endo-beta-N-acetylglucosaminidase H, non-sensitive). Chemically periodated 31 kDa protein showed statistically non-significant changes with human sparganosis sera by enzyme linked immunosorbent assay (ELISA). Therefore, the dominant epitopes of the 31 kDa molecule in human sparganosis were found to be mainly polypeptide, while N-glycans of the antigenic molecule in sparganum was minimal in anti-carbohydrate antibody production.
Animals ; Antigens, Helminth/*analysis/chemistry/immunology/metabolism ; Carbohydrates/analysis/immunology ; Chromatography, Affinity ; Chromatography, Ion Exchange ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Epitopes/analysis/immunology ; Glucosaminidase/metabolism ; Human ; Peptide-N4- (N-acetyl-beta-glucosaminyl) Asparagine Amidase/metabolism ; Periodic Acid/chemistry ; Sparganosis/*parasitology ; Sparganum/immunology/metabolism ; Spirometra/immunology/*metabolism ; Support, Non-U.S. Gov't

The cysteine proteases of Paragonimus westermani metacercariae are involved in metacercarial excystment, host immune modulation, and possibly in tissue penetration. In order to clarify the origin of the enzymes, 28 and 27 kDa cysteine proteases in metacercarial excretory-secretory products were purified through the FPLC system using Mono Q column chromatography. The polyclonal antibodies to the enzymes were produced in BALB/c mice. Immunolocalization studies revealed that both cysteine proteases were distributed at the linings of excretory bladder and excretory concretions of the metacercariae. It was suggested that the excretory epithelium of P. westermani undertake the secretory function of metacercarial cysteine proteases, in addition to its role as a route for eliminating waste products.
Animals ; Chromatography, Liquid ; Computational Biology ; Cysteine Endopeptidases/analysis/isolation & purification/*physiology ; Immunohistochemistry ; Mice ; Mice, Inbred BALB C ; Paragonimus/anatomy & histology/*enzymology ; Support, Non-U.S. Gov't

The plasmepsins are the aspartic proteases of malaria parasites. Treatment of aspartic protease inhibitor inhibits hemoglobin hydrolysis and blocks the parasite development in vitro suggesting that these proteases might be exploited their potentials as antimalarial drug targets. In this study, we determined the genetic variations of the aspartic proteases of Plasmodium vivax (PvPMs) of wild isolates. Two plasmepsins (PvPM4 and PvPM5) were cloned and sequenced from 20 P. vivax Korean isolates and two imported isolates. The sequences of the enzymes were highly conserved except a small number of amino acid substitutions did not modify key residues for the function or the structure of the enzymes. The high sequence conservations between the plasmepsins from the isolates support the notion that the enzymes could be reliable targets for new antimalarial chemotherapeutics.
Amino Acid Sequence ; Animals ; Aspartic Endopeptidases/*genetics ; Base Sequence ; Cloning, Molecular ; Conserved Sequence ; DNA, Protozoan/chemistry/genetics ; Human ; Molecular Sequence Data ; Plasmodium vivax/*enzymology/genetics/isolation & purification ; Polymerase Chain Reaction ; Sequence Alignment ; Sequence Analysis, DNA ; Support, Non-U.S. Gov't

In fascioliasis, T-helper 2 (Th2) responses predominate, while little is known regarding early immune phenomenon. We herein analyzed early immunophenotype changes of BALB/c, C57BL/6, and C3H/He mice experimentally infected with 5 Fasciola hepatica metacercariae. A remarkable expansion of CD19+ B cells was observed as early as week 1 post-infection while CD4+/CD8+ T cells were down-regulated. Accumulation of Mac1+ cells with time after infection correlated well with splenomegaly of all mice strains tested. The expression of tumor necrosis factor (TNF)-alpha mRNA in splenocytes significantly decreased while that of IL-4 up-regulated. IL-1beta expression was down-modulated in BALB/c and C57BL/6 mice, but not in C3H/He. Serum levels of transforming growth factor (TGF)-beta were considerably elevated in all mice during 3 weeks of infection period. These collective results suggest that experimental murine fascioliasis might derive immune suppression with elevated levels of TGF-beta and IL-4 during the early stages of infection.
Animals ; B-Lymphocytes/immunology ; CD4-Positive T-Lymphocytes/immunology ; CD8-Positive T-Lymphocytes/immunology ; Down-Regulation ; Fasciola hepatica/*immunology ; Fascioliasis/*immunology ; Immunophenotyping ; Immunosuppression ; Interleukin-4/blood/genetics/*immunology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C3H ; Mice, Inbred C57BL ; Spleen/immunology ; Transforming Growth Factor beta/blood/genetics/*immunology

PURPOSE: This study was designed to evaluate the clinical and radiographical results of anatomical reconstruction by Chen method for chronic lateral ankle instability. MATERIALS AND METHODS: Fifteen patients with chronic lateral ankle instability who had undergone anatomical reconstruction of anterior talofibular and calcaneofibular ligaments by Chen method were evaluated retrospectively. Average age of the patients was 31.3 years, and average follow-up period was 15.5 months. Preoperative and postoperative radiographs including varus stress view and magnetic resonance imaging (MRI) were analyzed. The clinical evaluation was performed according to the American Orthopaedic Foot and Ankle Society (AOFAS) scale. RESULTS: Radiographically average talar tilt angle was 15.3degrees preoperatively, and the difference with contralateral normal side was 10.1degrees. At last follow up, talar tile angle and the difference with contralateral side improved to 5.9degrees and 1.3degrees respectively. AOFAS scale was 66.6 preoperatively and 87.3 postoperatively. In MRI findings, four patients had associated intra-articular lesion such as articular cartilage defect, synovitis and osteoarthritis. The talar tilt angle improvement and AOFAS scale of patients without intra-articular lesion was better than those of four patients with intra-articular lesions. Surgical wound pain occurred in six patients and sural neuropathy in three patients. CONCLUSION: The anatomical reconstruction by Chen method was an easy and effective procedure for symptomatic chronic lateral ankle instability. Careful operative technique may prevent the surgical wound pain and sural neuropathy.
Animals ; Ankle ; Cartilage, Articular ; Follow-Up Studies ; Foot ; Humans ; Ligaments ; Magnetic Resonance Imaging ; Organic Chemicals ; Osteoarthritis ; Retrospective Studies ; Synovitis

The injury of popliteal artery in association with a total knee arthroplasty is rare with the incidence reported 0.03% to 0.05%, but the result of popliteal artery occlusion is extremely poor. Preexisting peripheral arterial disease, flexion deformity, tourniquet use, inadequate procedure during TKA, close proximity of popliteal artery to posterior cortex of tibia plateau, all of these are likely to increase the risk of the injury of popliteal artery during a knee replacement operation. We have experienced a patient who developed an arterial complication in association with a total knee arthroplasty and report the treatment as well as the discussion about the mechanisms of injury of popliteal artery.
Arthroplasty* ; Arthroplasty, Replacement, Knee* ; Congenital Abnormalities ; Humans ; Incidence ; Knee ; Peripheral Arterial Disease ; Popliteal Artery* ; Tibia ; Tourniquets

PURPOSE: To evaluate the midterm clinical and histological results after autologous chondrocyte implantation (ACI) for an articular cartilage defect of the distal femoral condyle. MATERIALS AND METHODS: Twenty four cases with an articular cartilage defect (Outerbridge grade IV) of the femoral condyle that was confirmed by MRI and the arthroscopic findings underwent ACI. Their mean age at the time of surgery was 42.8 years and the mean follow-up period was 53.2 months (range, 20-82 months). At the last follow up, the articular cartilage view (SPGR) of MRI was examined and the clinical results were evaluated using the HSS and Lysholm scores. In 8 cases, second-look arthroscopy and biopsy were performed and evaluated using histological and histochemical methods. RESULTS: All cases except for one showed well-regenerated articular cartilage on MRI. All cases showed significant clinical improvement in the HSS and Lysholm scores (p<0.0001), with the exception of the Lysholm score of an articular cartilage fracture. Histologically, the regenerated tissue appeared to be a hyaline-like cartilage in all specimens. CONCLUSION: ACI for the treatment of articular cartilage defects of the distal femoral condyle showed a good clinical and MRI results. In OA, the clinical results were relatively acceptable after an associated high tibial valgus osteotomy. However, a longer term follow-up study will be needed to reach a final conclusion.
Arthroscopy ; Biopsy ; Cartilage ; Cartilage, Articular ; Chondrocytes ; Follow-Up Studies ; Osteotomy

Fibrous pseudotumor of the testicular tunics is uncommon lesion. They typically arise as painless scrotal masses that may be associated with a hydrocele or history of trauma or infection. Two-thirds involve the tunica vaginalis testis, with infrequent involvement of other scrotal structures. Once excised, these lesions behave in a benign fashion. Typically, these masses are multinodular, but in rare cases they are diffuse, band-like myofibroblastic proliferations that encase the testis. We report here on a case of fibrous pseudotumor of the tunica vaginalis in 76 year-old patient. (Korean J Urol 2006;47:1130-1132)
Aged ; Humans ; Myofibroblasts ; Testis