This study was conducted to investigate whether administration of IH901, a ginseng intestinal metabolite, ameliorates exercise-induced oxidative stress while preserving antioxidant defense capability in rat skeletal muscles and lung. Eight adult male Sprague-Dawley rats per group were randomly assigned to the resting control, exercise control, resting with IH901 (25, 50, and 100 mg/kg) consumption (R/IH901), or exercise with IH901 (25, 50, and 100 mg/kg) consumption (E/IH901) group. The trained groups ran 35 min 2 days/week for 8 weeks. To analyze the IH901-training interaction, serum biochemical analysis, lipid peroxidation, citrate synthase, protein oxidation, antioxidant and superoxide dismutase in skeletal muscles and lung tissue were measured. Compared to the exercise control group, animals that consumed IH901 had significantly increased exercise endurance times (p < 0.05) and decreased plasma creatine kinase and lactate dehydrogenase levels (p < 0.05), while those in the E/IH901 groups had increased citrate synthase and anti-oxidant enzymes and decreased lipid peroxidation and protein oxidation (p < 0.05). In conclusion, IH901 consumption in aging rats after eccentric exercise has beneficial effects on anti-inflammatory and anti-oxidant activities through down-regulation of pro-inflammatory mediators, lipid peroxidation, and protein oxidation and up-regulation of anti-oxidant enzymes.
Aging ; Animals ; Antioxidants/administration & dosage/*pharmacology ; Dose-Response Relationship, Drug ; Lung/*drug effects/metabolism ; Male ; Muscle, Skeletal/*drug effects/metabolism ; Oxidative Stress/*drug effects ; Panax/chemistry ; Physical Conditioning, Animal ; Rats ; Rats, Sprague-Dawley ; Sapogenins/administration & dosage/blood/*metabolism/*pharmacology ; Specific Pathogen-Free Organisms

BACKGROUND: Panel reactive antibody (PRA) test is important in that it could minimize the errorneous report of HLA crossmatch in emergent situation and be used as a information on nationwide organ sharing. However, only a few laboratories manage to do this test by home-ade tray. This study was intended to resolve the cause of discrepancy between EIA and AHG-DC which would be encountered when supplement AHG-DC was tested for EIA positive samples. METHODS: Reactivity of Lambda Antigen Tray class I and class II mixed (LAT-, One Lambda, CA, USA) was evaluated to 23 specimens which had been positive in AHG phase of HLA crossmatch. All the samples for PRA were tested by EIA screening kit primarily and then AHG-DC was applied only in EIA positive samples. Samples showing discrepant results between two tests were further evaluated by EIA identification panel, platelet absorption test, dithiothreitol (DTT) treatment, and HLA crossmatch (AHG-DC and flowcytometry) with lymphocytes having antigen specificity reactive to antibodies identified by EIA kit. RESULTS: Of 23 samples, 21 (91.3%) showed strong reactivity to EIA and remaining 2 were confirmed to have IgM type alloantibodies. Of 92 samples for PRA, 22 (23.9%) were positive in anti-LA class I. 10 of 22 samples with positive EIA showed negative results by AHG-DC. While three of them were identified of their antibody specificity, of which 2 samples confirmed to have CYNAP (cytotoxicity negative, adsorption positive) antibodies and one sample to be nonspecific reaction to class I antigens, remaining 7 samples were negative to all the wells of EIA identification (id.) kit. Signal/cut-ff (S/C) of all three samples reactive to EIA id. kit were more than 1.75. CONCLUSIONS: HLA antibody screening by EIA from transplant candidate is considered to be appropriate in that EIA could detect IgG anti-LA of all tested sera and CYNAP antibodies as well. It needs further study with larger number of samples whether S/C of EIA would be informative in cases showing EIA positive/ AHG-DC negative.
Absorption ; Adsorption ; Antibodies ; Antibody Specificity ; Blood Platelets ; Dithiothreitol ; Histocompatibility Antigens Class I ; Immunoenzyme Techniques* ; Immunoglobulin G ; Immunoglobulin M ; Isoantibodies ; Lymphocytes ; Mass Screening ; Sensitivity and Specificity

BACKGROUND: Panel reactive antibody (PRA) test is important in that it could minimize the errorneous report of HLA crossmatch in emergent situation and be used as a information on nationwide organ sharing. However, only a few laboratories manage to do this test by home-ade tray. This study was intended to resolve the cause of discrepancy between EIA and AHG-DC which would be encountered when supplement AHG-DC was tested for EIA positive samples. METHODS: Reactivity of Lambda Antigen Tray class I and class II mixed (LAT-, One Lambda, CA, USA) was evaluated to 23 specimens which had been positive in AHG phase of HLA crossmatch. All the samples for PRA were tested by EIA screening kit primarily and then AHG-DC was applied only in EIA positive samples. Samples showing discrepant results between two tests were further evaluated by EIA identification panel, platelet absorption test, dithiothreitol (DTT) treatment, and HLA crossmatch (AHG-DC and flowcytometry) with lymphocytes having antigen specificity reactive to antibodies identified by EIA kit. RESULTS: Of 23 samples, 21 (91.3%) showed strong reactivity to EIA and remaining 2 were confirmed to have IgM type alloantibodies. Of 92 samples for PRA, 22 (23.9%) were positive in anti-LA class I. 10 of 22 samples with positive EIA showed negative results by AHG-DC. While three of them were identified of their antibody specificity, of which 2 samples confirmed to have CYNAP (cytotoxicity negative, adsorption positive) antibodies and one sample to be nonspecific reaction to class I antigens, remaining 7 samples were negative to all the wells of EIA identification (id.) kit. Signal/cut-ff (S/C) of all three samples reactive to EIA id. kit were more than 1.75. CONCLUSIONS: HLA antibody screening by EIA from transplant candidate is considered to be appropriate in that EIA could detect IgG anti-LA of all tested sera and CYNAP antibodies as well. It needs further study with larger number of samples whether S/C of EIA would be informative in cases showing EIA positive/ AHG-DC negative.
Absorption ; Adsorption ; Antibodies ; Antibody Specificity ; Blood Platelets ; Dithiothreitol ; Histocompatibility Antigens Class I ; Immunoenzyme Techniques* ; Immunoglobulin G ; Immunoglobulin M ; Isoantibodies ; Lymphocytes ; Mass Screening ; Sensitivity and Specificity

Ribosomal protein L21 (RPL21) is a structural component of the 60S subunit of the eukaryotic ribosome. This protein has an important role in protein synthesis and the occurrence of hereditary diseases. Pig is a common laboratory model, however, to the best of our knowledge, its RPL21 gene has not been cloned to date. In this study, we cloned and identified the full-length sequence of the pig RPL21 gene for the first time. In addition, we examined its expression pattern and function by using overexpression or knockdown approaches. As a result, we obtained a 604 bp segment that contains a 483 bp open reading frame encoding 160 amino acids. The pig RPL21 gene is located in the “+” strand of chromosome 11, which spans 2167 bp from 4199792 to 4201958. Pig RPL21 protein has nine strands and two helices in its secondary structure. Pig RPL21 is predominantly expressed in ovary and lung, at lower levels in kidney, small intestine, and skin, and at the lowest levels in heart and liver. Furthermore, RPL21 expression is closely connected with cell proliferation and cell cycle arrest. The results are intended to provide useful information for the further study of pig RPL21.
Amino Acids ; Cell Cycle Checkpoints ; Cell Proliferation ; Chromosomes, Human, Pair 11 ; Clone Cells ; Cloning, Molecular* ; Female ; Gene Expression ; Genetic Diseases, Inborn ; Heart ; Intestine, Small ; Kidney ; Liver ; Lung ; Open Reading Frames ; Ovary ; Ribosomal Proteins* ; Ribosomes ; Skin ; Sus scrofa

BACKGROUND: Proper gating is important in flow cytometric assays of lymphocyte subsets. Forward light scatter (FSC)/side light scatter (SSC) gating requires application of a lymphocyte purity correction when lymphocyte purity is less than 95%. We compared 3 different gating methods to establish an accurate gating method appropriate for a T-lymphocyte subset assay of bronchoalveolar lavage (BAL) fluid. METHODS: Leukocyte numbers and subtypes in 31 BAL fluid samples were assessed manually and by using an automatic hematology analyzer. T-lymphocyte subsets (T cells, T helper/inducer cells [Th], and T suppressor/cytotoxic cells [Tc]) were assessed by flow cytometry. We compared 3 methods of lymphocyte gating: CD45/SSC gating (reference method), FSC/SSC gating, and FSC/SSC gating with application of a lymphocyte purity correction. Lymphocyte purity was determined by CD45/CD14 staining of BAL fluid. RESULTS: We observed a significant correlation between lymphocyte percentage and lymphocyte purity (r = 0.453, P = 0.011). T-cell results obtained using the reference method were not correlated with the results of the other 2 gating methods (r = 0.189 each, P = 0.308 for FSC/SSC gating and P = 0.310 for FSC/SSC gating with purity correction). Mean differences between the reference method and FSC/SSC gating (T cells: 14.4%, P = 0.002; Th cells: 7.7%, P = 0.006; Tc cells: 7.1%, P = 0.001) were greater than those between the reference method and FSC/SSC gating with purity correction (T cells: 12.1%, P = 0.004; Th cells: 1.7%, P = 0.608; Tc cells: 0.2%, P = 0.957). CONCLUSIONS: Lymphocyte purity correction after FSC/SSC gating improved the accuracy of Th- and Tc-cell measurements, but not T-cell measurements. CD45 is essential for lymphocyte gating in T-lymphocyte subset assays of BAL fluid.
Bronchoalveolar Lavage ; Bronchoalveolar Lavage Fluid ; Flow Cytometry ; Hematology ; Leukocyte Count ; Light ; Lymphocyte Subsets ; Lymphocytes ; T-Lymphocyte Subsets ; T-Lymphocytes

BACKGROUND: Clinical and Laboratory Standards Institute (CLSI) guidelines (H42-A2) recommend the "CD45/SSC" gating method for assays on lymphocyte subset enumeration and CD16 exclusion for assays enumerating NK cells. In contrast, the Flow Cytometry Checklist (06/17/2010) of the College of American Pathology does not recommend a specific lymphocyte gating method, but recommends the correction of lymphocyte subset results for lymphocyte gate purity. METHODS: We compared lymphocyte subset results of EDTA-treated blood from 102 patients with various diseases and 12 normal controls, using 3 lymphocyte gating methods (CD45/SSC, FSC/SSC, and lymphocyte gate purity correction after FSC/SSC gating), and assessed the proportion of CD56-/CD16+ NK cells within the total NK cell population. RESULTS: Lymphocyte gate purity increased as the percentage of lymphocytes increased. However, lymphocyte subsets that consistently showed high lymphocyte gate purity could not be identified. The purity of the T cell population differed significantly depending on the gating method used: CD45/SSC vs. FSC/SSC, P=0.027; CD45/SSC vs. gate purity correction after FSC/SSC, P=0.002. However, the lymphocyte gate purity correction after FSC/SSC gating did not significantly improve the accuracy of the lymphocyte subset enumeration assay using FSC/SSC gating. The subset of CD56-CD16+ NK cells, constituted an average of 17.1% of total NK cells. Patients had higher proportions of CD56-CD16+ NK cells (13.1-25.5%) than did the normal controls (9.52%). CONCLUSIONS: In flow cytometric assays to evaluate lymphocytic subsets, the CD45 is inevitable for lymphocyte gating, whereas the measurement of CD16 is essential for the evaluation of NK cell proportions.
Checklist ; Flow Cytometry ; Humans ; Killer Cells, Natural ; Lymphocyte Subsets ; Lymphocytes

This study was performed to investigate the expression of two porcine endogenous retrovirus (PERV) elements, PERV gag and full-length conserved PERV, in blood cells collected periodically from organ-recipient monkeys that underwent pig to non-human primate xenotransplantation. The heart and kidney-respectively acquired from alpha-1,3-galactosyltransferase knockout (GT-KO) pigs that survived for24 and 25 days-were xenografted into cynomolgus monkeys. The two PERV elements expressed in the xenografted GT-KO pig organs were not present in the blood cells of the recipient monkeys. In the present study, we deduced that PERVs are not transmitted during GT-KO pig to monkey xenotransplantation.
Blood Cells ; Endogenous Retroviruses* ; Haplorhini* ; Heart ; Heterografts ; Macaca fascicularis ; Primates ; Swine ; Transplantation, Heterologous*

Background: Light-chain amyloidosis (AL) is the most common form of systemic amyloidosis. This study aimed to evaluate the usefulness of laboratory tests for light-chain clonality and bone marrow (BM) findings in AL amyloidosis. Methods: We retrospectively enrolled patients newly diagnosed with AL amyloidosis on pathological examination who underwent a BM biopsy. Laboratory test data for light-chain clonality were collected and compared. Amyloid deposits were identified with H&E, Congo red, and PAS stains. Results: We reviewed 98 patients with AL amyloidosis. Light chain clonality (λ, 64 cases; κ, 34 cases) was detected by serum immunofixation electrophoresis (IFE) (63.3%), urine IFE (70.8%), serum protein electrophoresis (PEP) (44.9%), urine PEP (44.8%), serum free light chain (SFLC) ratio (79.5%), and BM immunohistochemistry (IHC) (85.7%). Flow cytometric (FCM) assay identified aberrant BM plasma cells in 92.9% of cases. BM amyloid deposits were identified in 35 of the 98 cases (35.7%); 71.4% (25/35) were Congo red-positive, and 100.0% (35/35) were PAS-positive. Conclusion Laboratory tests for detecting light-chain clonality in AL amyloidosis in order of sensitivity include FCM assay for aberrant plasma cells, IHC for light chains on BM biopsy or clot section, SFLC ratio, and serum and urine IFE. Congo red staining of BM samples remains an important tool for identifying amyloid deposits in BM. Periodic acid-Schiff (PAS) staining can be useful in diagnosing some cases of Congo red-negative amyloidosis.