Xenotransplantation using porcine organs could potentially associate with the risk of pathogenic infections, because human tropic porcine endogenous retrovirus (PERV) particles could be released from pig cells or organs. While there is no evidence of PERV transmission to human, safety issues become a paramount concern. For the prevention of this transmission, specific immunological tools must be provided for PERV transmission detection. In this study we described the expression of PERV envelope proteins and the production of a specific antibody against PERV envelope (Env) glycoprotein. The nucleotide sequence harboring the partial region of glycoprotein 70 was cloned into the pET vector and envelope protein was expressed in E. coli. Approximately 42 kDa recombinant Env protein (PERV Env-aa357) was purified by the Ni-affinity column. For antibody production, mice were immunized with the recombinant PERV Env-aa357. The generated anti-serum was tested using Western blot and immunocytochemical assay. We found that anti-PERV Env serum displayed the specificity against the PERV Envs (PERV-A and PERV-B) expressed not only in E. coli but also in mammalian cells, and PERV particles within the porcine cell lines (PK 15 and PK-1). Taken together, PERV antibody could be useful for detecting PERV infection or xenotransplantation transmission.
Animals ; Antibody Formation ; Base Sequence ; Blotting, Western ; Cell Line ; Clone Cells ; Endogenous Retroviruses ; Gene Products, env ; Glycoproteins ; Humans ; Mice ; Proteins ; Sensitivity and Specificity ; Transplantation, Heterologous

PURPOSE: To report upon two cases of anterior capsule opacification (ACO) proliferating into the pupillary region of the anterior surface of a single piece acrylic AcrySof(R) intraocular lens (IOL) (SA60AT, Alcon Laboratories, Inc., USA). The surgical removal of the ACO was also reported. METHODS: An 80-year-old woman and a 75-year-old woman underwent phacoemulsification with implantation of a single piece foldable AcrySof(R) IOL in the posterior capsular bag. ACO proliferating from the opposite sides of the IOL along the central axis were found three months after surgery. To recover visual acuity in the 80-year-old woman's eye, ACO was surgically removed with a Sinskey hook and irrigation and aspiration system. RESULTS: The patient's visual acuity was recovered after surgical removal of the ACO. Histopathologic findings of the removed tissues revealed fibroblastic proliferation of lens epithelial cells. The ACO in 75-year-old woman's eye was not treated, as it was not associated with a visual disturbance. CONCLUSIONS: In eyes containing an AcrySof(R) IOL, clinically significant ACO proliferation into the pupillary region along the central axis can occur. Surgical removal can be beneficial for recovery of visual acuity.
Aged ; Aged, 80 and over ; Axis, Cervical Vertebra ; Capsule Opacification ; Epithelial Cells* ; Female ; Fibroblasts* ; Humans ; Lenses, Intraocular* ; Phacoemulsification ; Visual Acuity

Xenotransplantation, as a potential solution to the shortage of human organs, is associated with a number of concerns including immunologic rejection and xenogenic infection. While the pigs are considered the most suitable organ source for xenotransplantation, there is a potential public health risk due to zoonosis. Among the known porcine zoonotic microbes, Porcine Endogenous Retrovirus (PERV) is the most considerable virus. PERV belongs to the Gammaretrovirus and has been divided into three groups (A, B, and C). To characterize the gag of PERVs, we isolated the genomic DNAs from three pig breeds (Birkshire, Duroc, and Yorkshire) and two types of SPF miniature pigs. About 1.5 kb fragments covering full length of gag were amplified and cloned into T-vector. A total of 38 clones were obtained and sequenced. Nucleotide sequences were analyzed and phylogenetic trees were constructed from the nucleotide and deduced amino acids. PERV-A, -B and -C were present in the proportion of 47, 19 and 34%, respectively. Regardless of origin or subgroups, gag clones showed highly homology in nucleotide and deduced amino acid sequences. Deduced amino acids sequence alignments showed typical conserve sequences, Cys-His box and processing sites. Among analyzed clones, about 28% of isolates had the correct open reading frame. To test the functional expression of Gag protein, gag was subcloned into expression vector and confirmed its expression in HeLa cell. This research provides the fundamental information about molecular characteristics of gag gene and functional Gag protein related xenotropic PERVs.
Amino Acid Sequence ; Amino Acids ; Base Sequence ; Clone Cells ; DNA ; Endogenous Retroviruses* ; Gammaretrovirus ; Gene Products, gag ; Genes, gag* ; HeLa Cells ; Humans ; Korea* ; Open Reading Frames ; Public Health ; Sequence Alignment ; Swine* ; Transplantation, Heterologous

OBJECTIVE: Comparison of protein expressions by two-dimensional gel electrophoresis (2-DE) in normal cervix and squamous cell carcinoma tissues in Korean women. METHODS: Normal cervix and squamous cell carcinoma tissues were solubilized with 2-DE buffer and the first dimension of PROTEAN IEF CELL, isoelectric focusing (IEF), was performed using pH3-10 linear IPG strips of 17 cm. And then running 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and sliver stain. Scanned image was analyzed using PDQuest 2-D softwareTM. Protein spot spectrum was identified by assisted laser desorption/ionization-time of fighting (MALDI-TOF) and the protein mass spectrum identifications were performed by searching protein databases of Swiss-prot/TrEMBL, Mascot and MS-FIT. RESULTS: We found 9 up-regulation proteins (Alpha enolase, Keratin 19 type I, Keratin 20 type I, Keratin 13 type I, beta-actin, Aflatoxin B1 aldehyde reductase 1, Annexin A2, Squamous cell carcinoma antigen 2, unknown), 7 down-reguation proteins (Annexin 1, Myosin regulatory light chain 2, 14-3-3 protein epsilon, Heat shock 27 kDa protein, Hypothetical protein (DKFZP434C1715), Tumor necrosis factor receptor superfamily member 13B, Smoth muscle protein 22-alpha) and 6 up and down-regulation proteins (Tropomyosin 1, Tropomyosin 2, Tropomyosin 3, Serine (or cysteine) proteinase inhibitor, Phosphatidylinositol transfer protein alpha isoform, Src homology 3 domain-containing protein HIP-55) between normal cervix and squamous cell carcinoma cell tissues. CONCLUSION: 2-DE offers total protein expressions between normal cervix and squamous cell carcinoma cell tissues, and searching of differently expressed protein for the diagnostic markers of squamous cell carcinoma tissue.
14-3-3 Proteins ; Actins ; Aflatoxin B1 ; Aldehyde Reductase ; Annexin A2 ; Carcinoma, Squamous Cell ; Cervix Uteri ; Databases, Protein ; Down-Regulation ; Electrophoresis, Gel, Two-Dimensional* ; Electrophoresis, Polyacrylamide Gel ; Female ; Hot Temperature ; Humans ; Isoelectric Focusing ; Keratin-13 ; Keratin-19 ; Keratin-20 ; Mass Spectrometry* ; Muscle Proteins ; Myosin Light Chains ; Phospholipid Transfer Proteins ; Phosphopyruvate Hydratase ; Receptors, Tumor Necrosis Factor ; Running ; Serine ; Shock ; Sodium Dodecyl Sulfate ; Tropomyosin ; Up-Regulation ; Uterine Cervical Neoplasms*

The Aspergillus species produces metabolic products that play a significant role in the destructive processes in the lung. We experie nced a case of chronic necrotizing pulmonary aspergillosis caused by an Aspergillus niger infection, which contained numerous calcium oxalate crystals in the necrotic lung tissue. A 46-year-old man, who had a history of pulmonary tuberculosis, presented with high fever, intermittent hemoptysis and pulmonary infiltrations with a cavity indicated by the chest radiograph. Despite being treated with several antibiotics and anti-tuberculosis regimens, the high fever continued. The sputum cultures yielded A. niger repeatedly, and intravenous amphotericin B was then introduced. The pathological specimen obtained by a transbron chial lung biopsy revealed numerous calcium oxalate crystals in a background of acute inflam matory exudates with no identification of the organism. Intravenous amphotericin B was con tinued at a total dose of 1600 mg, and at that time he was afebrile, although the intermittent hemoptysis continued. On the 63rd hospital day, a massive hemoptysis (about 800 mL) developed, which could not be controlled despite embolizing the left bronchial artery. He died of respiratory failure the next day. It is believed that the oxalic acid produced by A. niger was the main cause of the patient's pulmonary injury and the ensuing massive hemoptysis.
Amphotericin B ; Anti-Bacterial Agents ; Aspergillus niger* ; Aspergillus* ; Biopsy ; Bronchial Arteries ; Calcium Oxalate ; Exudates and Transudates ; Fever ; Hemoptysis ; Humans ; Hyperoxaluria* ; Invasive Pulmonary Aspergillosis ; Lung ; Lung Injury ; Middle Aged ; Niger ; Oxalic Acid ; Radiography, Thoracic ; Respiratory Insufficiency ; Sputum ; Tuberculosis, Pulmonary

Bronchial artery embolization (BAE) is a well accepted and effective treatment for massive and recurrent hemoptysis. However, several complications of BAE have been reported. Cortical blindness is defined as a loss of vision caused by bilateral occipital lobe lesions with normal pupillary light reflexes and a normal fundus. The reported incidence of transient cortical blindness (TCB) after cerebrovertebral angiography is approximately 1%. Two cases of TCB after BAE were found from a Medline search. Here, we report another case of TCB who was treated with BAE for a massive hemoptysis.
Incidence

Bronchial artery embolization (BAE) is a well accepted and effective treatment for massive and recurrent hemoptysis. However, several complications of BAE have been reported. Cortical blindness is defined as a loss of vision caused by bilateral occipital lobe lesions with normal pupillary light reflexes and a normal fundus. The reported incidence of transient cortical blindness (TCB) after cerebrovertebral angiography is approximately 1%. Two cases of TCB after BAE were found from a Medline search. Here, we report another case of TCB who was treated with BAE for a massive hemoptysis.
Incidence

OBJECTIVE: To know the effect of adenosine 5'-triphosphate (ATP) on intracellular calcium level and cell proliferation in cervical cancer cells. METHODS: Study design: Four different human cervical cancer cell lines (Caski, C33A, HeLaS3 and SiHa) were used in this study. The change of intracellular calcium level, cell proliferation and the activity of proliferation- and calcium-related transcription factors by extracellular ATP were examined in these cell lines. RESULTS: Extracellular ATP induced calcium mobilization, cell proliferation and the activation of NF-kappa B in all cell lines used. CONCLUSION: These results suggest that calcium mobilization and NF-kappa B dependent signaling pathway play an important role in the cell proliferation by ATP in cervical cancer.
Adenosine Triphosphate ; Adenosine* ; Calcium* ; Cell Line ; Cell Proliferation ; Humans ; NF-kappa B* ; Transcription Factors ; Uterine Cervical Neoplasms*

Four trials of external quality assessment in diagnostic hematology were performed in 2005 with about 500 participating laboratories in Korea. We performed quality assessment for white blood cell count, hemoglobin, red blood cell count, platelet count, white cell differential count, red blood cell morphology. The response rate was more than 97%. The coefficients of variation in hemoglobin and RBC number was stable but variable in platelet number and WBC number according to measuring cell counts. Test results showed wide variation according to measuring machine and reagents.
Cell Count ; Erythrocyte Count ; Erythrocytes ; Hematology* ; Indicators and Reagents ; Korea* ; Leukocyte Count ; Platelet Count

PURPOSE: The molecular pathology of cervical cancers associated with human papillomavirus infection is presently unclear. In an effort to clarify this issue, the gene expression profiles and pathogenic cellular processes of cervical cancer lesions were investigated. MATERIALS AND METHODS: Cervical cancer biopsies were obtained from patients at the Department of Obstetrics and Gynecology, The Catholic University of Korea. The disease status was assigned according to the International Federation of Gynecology and Obstetrics. The tissue samples of 11 patients (invasive cancer stage Ib- IIIa) were investigated by a cDNA microarray of 4, 700 genes, hierarchical clustering and the Gene Ontology (GO). Total RNA from cervical cancer and non-lesional tissues were labeled with Cy5 and Cy3. The HaCaT human epithelial keratinocyte cell line was used as a negative control cell. The stages of invasive cancer were Ib to IIIb. All specimens were obtained by punch-biopsies and frozen in liquid nitrogen until required. RESULTS: 74 genes, showing more than a 2 fold difference in their expressions, were identified in at least 8 of the 11 patients. Of these genes, 33 were up-regulated and 41 were down-regulated. The gene expression profiles were classified into 345 mutually dependent function sets, resulting in 611 cellular processes according to their GO. The GO analysis showed that cervical carcinogenesis underwent complete down-regulation of cell death, protein biosynthesis and nucleic acid metabolism. The genes related to nucleic acid binding and structural molecule activity were also significantly down-regulated. In contrast, significant up-regulation was shown in the skeletal development, immune response and extracellular activity. CONCLUSION: These data are suggestive of potentially significant pathogenetic cellular processes, and showed that the down-regulated functional profiling has an important impact on the discovery of pathogenic pathways in cervical carcinogenesis. GO analysis can also overcome the complexity of the expression profiles of the cDNA microarray via a cellular process level approach. Thereby, a valuable prognostic candidate gene, with real relevance to disease-specific pathogenesis, can be found at the cellular process levels.
Biopsy ; Carcinogenesis ; Cell Death ; Cell Line ; DNA, Complementary* ; Down-Regulation ; Gene Expression* ; Gene Ontology ; Gynecology ; Humans ; Keratinocytes ; Korea ; Metabolism ; Nitrogen ; Obstetrics ; Oligonucleotide Array Sequence Analysis* ; Papillomavirus Infections ; Pathology, Molecular ; Protein Biosynthesis ; RNA ; Transcriptome* ; Up-Regulation ; Uterine Cervical Neoplasms*