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

Four trials of external quality assessment in diagnostic hematology were performed in 2008 with average 822 participating laboratories in Korea. We performed quality assessment for white blood cell count, hemoglobin, hematocrit, red blood cell count, platelet count, blood cell morphology, prothrombin time and activated partial thromboplastin time. The response rate was more than 96.5%. The coefficients of variation in hemoglobin, hematocrit and RBC was stable but variable in platelet count and WBC count according to measuring cell count. Test results of blood cell morphology showed variation among various cell morphologies.
Blood Cells ; Cell Count ; Erythrocyte Count ; Hematocrit ; Hematology ; Hemoglobins ; Korea ; Leukocyte Count ; Partial Thromboplastin Time ; Platelet Count ; Prothrombin Time

Four trials of external quality assessment in diagnostic hematology were performed in 2007 with average 722 participating laboratories in Korea. We performed quality assessment for white blood cell count, hemoglobin, hematocrit, red blood cell count, platelet count, blood cell morphology, prothrombin time and activated partial thromboplastin time. The response rate was more than 95.2%. The coefficients of variation in hemoglobin, hematocrit and RBC were stable but variable in platelet count and WBC count according to measuring cell counters. Test results of blood cell morphology showed variation among various cell morphologies.
Blood Cells ; Cell Count ; Erythrocyte Count ; Hematocrit ; Hematology ; Hemoglobins ; Korea ; Leukocyte Count ; Partial Thromboplastin Time ; Platelet Count ; Prothrombin Time

Four trials of external quality assessment in diagnostic hematology were performed in 2007 with average 722 participating laboratories in Korea. We performed quality assessment for white blood cell count, hemoglobin, hematocrit, red blood cell count, platelet count, blood cell morphology, prothrombin time and activated partial thromboplastin time. The response rate was more than 95.2%. The coefficients of variation in hemoglobin, hematocrit and RBC were stable but variable in platelet count and WBC count according to measuring cell counters. Test results of blood cell morphology showed variation among various cell morphologies.
Blood Cells ; Cell Count ; Erythrocyte Count ; Hematocrit ; Hematology ; Hemoglobins ; Korea ; Leukocyte Count ; Partial Thromboplastin Time ; Platelet Count ; Prothrombin Time

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

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

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: 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*

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*

PURPOSE: Paclitaxel and cisplatin, active drugs in the treatment of non-small-cell lung cancer (NSCLC), have been found to be synergistic and less myelotoxic in combination when the paclitaxel is given 24 hr prior to the cisplatin. Their antitumor activity and toxicity in patients with advanced NSCLC has been evaluated herein. MATERIALS AND METHODS: Seventy-four chemonaive patients, with advanced NSCLC, were enrolled. Paclitaxel, 175 mg/m2, was administered on day 1, followed 24 hr later by cisplatin, 75 mg/m2, on day 2. RESULTS: The overall response rate, median time to progression and median survival time were 51%, 7.1 months (95% confidence interval (CI), 5.5~8.7 months) and 13.7 months (95% CI, 11.3~16.1 months), respectively. There were significant differences in the overall survival rates in relation to stage and the ECOG performance status(PS). The toxicity was mainly nonhematological. Grade > or =3 neuropathy occurred in 2 (3%) patients, myalgia in 3 (4%), and bone pain in 3 (4%). The hematological toxicity was mild, and no grade 3 or 4 neutropenia was observed. CONCLUSION: The combination of paclitaxel and cisplatin is an effective and tolerable treatment regimen for advanced NSCLC during first line chemotherapy. The main toxicity was nonhematological, such as peripheral neuropathy, myalgia and bone pain, whereas the hematological toxicity itself was mild.
Cisplatin* ; Drug Therapy ; Drug Therapy, Combination* ; Humans ; Lung Neoplasms* ; Lung* ; Myalgia ; Neutropenia ; Paclitaxel* ; Peripheral Nervous System Diseases ; Survival Rate