Biomedical subjects

C S Choi

Publications and source records attributed to C S Choi.

At least 55 records · Page 3Linked to original sources

PubMed1997

Monitoring of porcine reproductive and respiratory syndrome virus infection in boars.

A major concern exists on transmission of porcine reproductive and respiratory syndrome virus (PRRSV) via semen and effect of vaccination on PRRSV shedding in semen. Recent reports suggest that the virus can be transmitted by semen from boars infected experimentally or from natural sources. Seminal shedding, viremia, and changes in semen quality in boars with or without vaccination were examined. Nine boars were divided into three groups (three boars/group). Group I boars were vaccinated with 2 ml of RespPRRS vaccine (NOBL Laboratory) intramusculary and groups II and III were non-vaccinated. At 28 post-vaccination study days, group I and group II boars were challenged with virulent PRRSV VR-2332 at 2 ml of 10(4.0) TCID50 per boar intranasally. Group III served as non-vaccinated and non-challenged control. Semen and serum samples were collected from -9 pre-vaccination study days to 85 post-challenge study days and tested for the presence of PRRSV by virus isolation and reverse transcription-nested polymerase chain reaction (RT-nPCR). Prior to detection of PRRSV RNA from samples, conditions for RT-nPCR were optimized. Two primer sets, an external and an internal, were selected for RT-nPCR. The first round of PCR using an external primer set could detect 10 TCID50 of PRRSV/reaction. However, nested PCR could detect as little as 0.01 TCID50 of PRRSV/reaction. PRRS vaccine virus was not isolated from vaccinated pigs, but the vaccine virus RNA was detected from three boars, at day 6 to 15, 9 to 12, and 15 to 21 post-vaccination by RT-nPCR. Following challenge, two of non-vaccinated/challenged boars shed virus into semen up to 50 and 57 days post-challenge, respectively. The group I vaccinated boars did not shed virus into semen after challenge. The non-vaccinated/challenged group featured sperm abnormalities in the form of significantly increased incidence of proximal droplets and abnormal tails at 36-50 days post-challenge. The latter defect was observed to increase similarly in vaccinated/challenged boars as well.

Animals↗

PubMed1997

Involvement of different second messengers in parathyroid hormone- and interleukin-1-induced interleukin-6 and interleukin-11 production in human bone marrow stromal cells.

Previous studies have suggested that increased secretion of bone active cytokines, such as interleukin-6 (IL-6) and interleukin-11 (IL-11), from osteoblasts and stromal cells play a pivotal role in the activation of osteoclasts and the genesis of osteoporosis. Various systemic and local factors can stimulate IL-6/IL-11 production, but the intracellular mechanism for such stimulation is largely unknown. In this study, we characterized the second messenger signaling in parathyroid hormone (PTH)- and IL-1-induced production of IL-6/IL-11 and studied the possible modulating effects of estrogen. rhPTH(1-34) and rhIL-1 alpha dose-dependently stimulated IL-6 and IL-11 production from human bone marrow stromal cells (hBMSCs). Agonists for protein kinase A (PKA) (forskolin), and protein kinase C (PKC) (phorbol 12-myristate 13-acetate; PMA) also stimulated IL-6/IL-11 production. Rp-diastereoisomer of adenosine cyclic 3',5'-phosphorothioate (Rp-cAMPS) and H-8, inhibitors of PKA, significantly inhibited PTH-stimulated IL-6/IL-11 production, but did not inhibit IL-1-stimulated IL-6/IL-11 production. In contrast, staurosporine and calphostin C, inhibitors of PKC, suppressed IL-1-stimulated, but not PTH-stimulated, IL-6/ IL-11 production. Pretreatment of cells with 17 beta-estradiol (17 beta-E2) antagonized IL-1-stimulated IL-6 production. However, PTH-stimulated IL-6 production and IL-1- and PTH-stimulated IL-11 production were not affected by 17 beta-E2. Similarly, 17 beta-E2 inhibited PMA-stimulated IL-6 production, whereas neither forskolin-stimulated IL-6/ IL-11 production nor PMA-stimulated IL-11 production was affected by 17 beta-E2. These results indicate that different second messengers are involved in PTH- and IL-1-induced IL-6 and IL-11 production by hBMSCs: PTH and IL-1 stimulate IL-6/IL-11 production via a PKA-dependent and PKC-dependent pathway, respectively. Furthermore, our results suggest that regulation of cytokine production by estrogen in hBMSCs is selective; only the IL-1-induced IL-6 production, which is mediated by PKC pathway, is inhibited, but PTH-induced IL-6 production and PTH/IL-1-induced IL-11 production are not inhibited by estrogen.

Bone Marrow↗

PubMed1996

Differential infection of porcine alveolar macrophage subpopulations by nonopsonized Mycobacterium bovis involves CD14 receptors.

The resurgence in mycobacterial infection worldwide has led to renewed attention to the pathogenesis of Mycobacterium species. The purpose of this study was to characterize the infection of alveolar macrophages (AMs) by nonopsonized Mycobacterium bovis, and to elucidate the mechanism by which a differential infection of subpopulations of AM may occur. A difference in susceptibility to Mycobacterium bovis infection of subpopulations of AMs was observed, such that the least dense cells were the least susceptible (21.4 +/- 10.7%) and the most dense cells were the most readily infected (61.8 +/- 5.6%). The percentage of AMs staining for CD14 receptors showed a similar differential distribution, with fewer of the least dense cells expressing CD14 and a greater percentage of the most dense cells staining for CD14 receptor expression. To investigate the role of CD14 receptors in the infection of AMs, anti-CD14 antibody was added to the cell cultures. Infection of AM by Mycobacterium bovis was blocked by up to 60.2% by anti-CD14 antibody but not by isotype control antibody. The results of this study suggest that Mycobacterium bovis selectively infects AM subpopulations, specifically those with the greatest expression of CD14, a putative receptor mechanism for Mycobacterium bovis infection of porcine AM.

Animals↗

PubMed1996

Structural polypeptides of the American (VR-2332) strain of porcine reproductive and respiratory syndrome virus.

The structural polypeptides of the isolate VR-2332 of porcine reproductive and respiratory syndrome virus were analyzed in sucrose gradient-purified virions. The virus had an average density of 1.15 g/cm3 and contained, by SDS-PAGE, three major polypeptides with apparent molecular weights of 15, 19 and 26-30 kDa, which were designated as nucleocapsid (N), matrix (M) and envelope (E), respectively. The predominant structural protein was N. N-glycosidase F digestion only affected E whereas O-glycosidase or endoglycosidase H digestion had no effect, suggesting that the viral glycoproteins contain only complex N-linked carbohydrates.

Arterivirus↗

PubMed1996

Effect of cholestyramine on the formation of pigment gallstone in high carbohydrate diet-fed hamsters.

This study was designed to investigate the effect of cholestyramine on the formation of pigment gallstones in high carbohydrate diet-fed hamsters and whether that effect occurred because of cholecystokinin action. Forty seven hamsters were divided into three groups: group I(n = 16) was fed on normal rodent chow(43% carbohydrate), group II(n = 14) was fed on a high CHO diet(65% carbohydrate), group III(n = 17) was fed on a high CHO diet containing 4% cholestyramine. Gallstones developed in 0% of group I, 42.9% of group II and 5.9% of group III(P < 0.05, group II vs III). To evaluate the chronic status of cholecystokinin level, the wet weight of pancreas and the average area of pancreatic acinar in microscopic high power field were measured. There was no significant difference between group II and group III in pancreatic weight and average area of pancreatic acinar(P > 0.05). In gallbladder bile analysis, there was also no significant difference between group II and group III in cholesterol, phospholipid, total calcium, total bilirubin and bile acid levels. In conclusion, cholestyramine decreases the frequency of pigment gallstone formation in high CHO diet-fed hamsters, but it is not clear whether the mechanism of cholestyramine decreasing the gallstone formation is due to the action of cholecystokinin.

Animals↗

PubMed1995

Colorimetric assay using XTT for assessing virulence of avian Pasteurella multocida strains.

A colorimetric assay using sodium 3,3'-[1[(phenylamino)carbonyl]3,4- tetrazolium]-bis(4-methoxy-6-nitro) benzene sulfonic acid hydrate (XTT) was adapted to quantitate bactericidal activity of chicken macrophage HD 11 cell line against five Pasteurella multocida strains and an avirulent transposon insertion mutant. The strains used were virulent P1059, and D92, and four avirulent strains including a streptomycin resistant mutant of P1059 (P1059 SmR), two live vaccine strains namely, the Clemson University (CU) and M9, and a transposon insertion mutant PmTn-294. Percentage of bacteria killed by chicken macrophage (HD 11) cells was determined by extrapolation from a standard formazan curve derived by incubating XTT with known bacterial cell numbers of each strain. The amount of formazan as measured by absorption at 450 nm was directly related to the number of viable bacterial cells. The percentages of P1059 SmR, CU, M9 and PmTn-294 killed by HD 11 cells were approximately 50%, 61%, 25% and 34%, respectively. By contrast, the virulent P1059 and D92 strains were resistant to killing, and were able to replicate inside the HD 11 cells. Association of virulence with resistance to phagocytic killing by HD 11 cells as assessed by the colorimetric bactericidal assay, was validated with resistance to complement (C')-mediated killing and a turkey mortality test. Strains P1059 and D92 were resistant to C'-mediated killing, whereas strains P1059 SmR, CU, M9 and PmTn-294 strains were susceptible. All turkeys challenged with P1059 or D92 were dead within 18 hrs. Mortality did not occur in turkeys challenged with strains of P1059 SmR, M9 and PmTn-294. The mortality among CU challenged turkeys ranged from 0 to 40%. The results suggest that the colorimetric bactericidal assay using XTT can be used to quantitate chicken macrophage phagocytic killing of P. multocida strains, and may be a valuable assay to differentiate virulent from avirulent strains of avian P. multocida.

Animals↗

PubMed1995

Gas flames, closed spaces, and hypoxia.

The investigation of the deaths of an elderly couple found in a small hotel room that was tightly sealed and heated by an open flame gas heater is presented. Autopsy and toxicologic studies did not reveal a definitive cause of death. Subsequent research and an experiment to duplicate the conditions support the contention that the cause of death was hypoxia due to oxygen depletion by the gas flame.

Aged↗

PubMed1995

IgA response in mice infected with Neodiplostomum seoulensis.

To observe the production of IgA in Balb/c mice with neodiplostomiasis, 20 mice were infected with each 200 metacercariae of Neodiplostomum seoulenis. Sera and the duodenums were obtained 3, 7, 14, 28 days post-infection (PI) from five mice each group. Neodiplostomum specific IgA in serum by the enzyme-linked immunosorbent assay increased from 7 days PI and persisted till 28 days PI. Immunohistochemistry for IgA was done with sections of the duodenum. The IgA-positive reaction was generally seen in the lamina propria and submucosa. Some of epithelial cells were positive at 7 and 14 days PI. The present finding showed that Neodiplostomun specific IgA antibody increased in serum and that there was local reaction of IgA in the mucosa and submucosa of the duodenum but not directly related with worm expulsion.

Animals↗

PubMed1994

Nitric oxide generation from streptozotocin.

Streptozotocin (STZ), a diabetogenic agent, is thought to damage pancreatic beta-cells by activating immune mechanisms and by alkylating DNA. In the present study, we demonstrated that STZ can produce nitric oxide (NO), a bioregulatory and cytotoxic molecule. When STZ was dissolved in a sodium phosphate buffer (50 mM, pH 7.4) and irradiated with a 22 W circular fluorescent light, nitrite and nitrate, stable oxidation products of NO, were produced. The wavelengths of light most responsible for the photo-decomposition were 300-310 nm and 410-420 nm. When a mixture of reduced hemoglobin and STZ was irradiated with UV light (280-320 nm), hemoglobin underwent characteristic NO-dependent spectral changes. STZ relaxed deendothelialized aortic strips only in the presence of light. STZ/light-dependent relaxation was attenuated by reduced hemoglobin. These results indicated photo-induced NO production from STZ. NO generation depended on the concentration of STZ, the duration of irradiation, and the distance between sample and light source. In acidic conditions, NO production from STZ was spontaneous even in the dark. Light-independent NO generation was augmented by increasing acidity, and markedly diminished in a D2O-based buffer, indicating the involvement of protons in the mechanism of STZ decomposition in acid. These results imply the usefulness of STZ as an NO-generating reagent, and indicate that direct NO-generation may be a mechanism of STZ toxicity in diabetogenesis.

Animals↗

PubMed1994

The missing link. An unusual case of embolization.

Embolization is a well-recognized phenomenon in medicine and forensic pathology and generally involves formed blood elements, bone, air, amniotic fluid, or more exotic items such as bullets. We report the embolization of a chain-saw link. The case demonstrates how, without complete autopsy and investigation, a penetrating sharp-force injury could be misdiagnosed and a false conclusion as to the commission of a crime could occur. Although the forensic literature is replete with examples of bizarre emboli, from bullets to suspected crystals of methylmethacrylate, a literature review failed to find a single case of penetrating chain-saw-link injury as a cause of traumatic death. Most reports of chain-saw incidents cite accidental injuries to hands and arms or dramatic facial injuries due to entrapment and recoil (1,2). One fatality occurred from a skull fracture and cerebral injury when a 19-year-old man fell onto a saw.

Aged↗

PubMed1993

Tumors in hepatobiliary tract and pancreatic islet tissues of transgenic mice harboring gastrin simian virus 40 large tumor antigen fusion gene.

Gastrin is expressed in the gastric antrum and in fetal pancreatic islets but not in adult islets. We have now identified the hepatobiliary tract as another, previously unknown, potential site of gastrin gene expression. Two human gastrin simian virus 40 large tumor antigen (SV40 T antigen) fusion genes containing 1.5 kb of 5' flanking sequence and 10.5 kb that included 5.5 kb upstream, 1.5 kb downstream, and the entire transcribed region were used to generate transgenic mice. Analysis of several transgenic lines, derived from both fusion genes, revealed development of transmissible hepatobiliary tract tumors and pancreatic islet cell tumors. Analysis of each of the tumor cells demonstrates expression of SV40 T antigen but no expression of gastrin. Of the two fusion genes, only the 10.5-kb sequence induces hyperplasia of gastrin-producing cells in the antrum. Analysis of these cells demonstrates expression of SV40 T antigen and gastrin, suggesting that the 10.5-kb sequence is sufficient for gastrin cell hyperplasia in the antrum. These data raise the possibility that gastrin is transiently expressed in the hepatobiliary tract.

Adenoma, Islet Cell↗

PubMed1993

Tissue tropisms of porcine parvovirus in swine.

Late-term gestation swine fetuses, similar to adult animals, are able to effectively mount immune response and survive porcine parvovirus (PPV) infection. An exception to this is the Kresse strain of PPV, which causes fetal death in late-term gestation swine fetuses. In an effort to understand the basis for this profound difference in pathogenicity between Kresse strain and the prototype strain of PPV, NADL-8, studies were designed to examine potential difference in sites of replication and quantity of virus produced between Kresse and NADL-8 strains. In order to define the sites of viral replication or sites of viral sequestration in situ hybridization, using digoxigenin-labeled strand-specific oligonucleotide probes, was applied to detect the presence of either double stranded viral DNA (RF) or viral single stranded DNA in tissues of infected fetuses. The presence and the state of viral DNA was confirmed by Southern blot hybridization. Relative amounts of RF-DNA synthesis in each tissue was compared between the two virus strains. Virus replication appeared to be of comparable levels in the livers of both NADL-8 and Kresse infected swine fetuses. Differences between these strains were observed in the brain and spleen; RF-DNA was detected in the brain and spleens of Kresse infected fetuses but not in NADL-8 infected ones. These findings indicate that differences in DNA replication of the PPV strains in selective sites, as well as the quantity of virus produced, may explain the distinction in pathogenesis of these viruses.

Animals↗

PubMed1993

Pathogenesis of porcine reproductive and respiratory syndrome virus infection in mid-gestation sows and fetuses.

Two experiments were undertaken to evaluate whether porcine reproductive and respiratory syndrome (PRRS) virus was able to cross the placenta and infect midgestation fetuses following intranasal inoculation of sows and whether PRRS virus directly infected fetuses following in utero inoculation. In experiment 1, eight sows between 45 and 50 days of gestation were intranasally inoculated with PRRS virus (ATCC VR-2332), and four control sows were inoculated with uninfected cell culture lysate. Virus inoculated sows were viremic on postinoculation (PI) days 1, 3, 5, 7 and 9, shed virus in their feces and nasal secretions, and became leukopenic. Sixty-nine of 71 fetuses from principal sows euthanized on PI day 7, 14 or 21 were alive at necropsy and no virus was isolated from any of the fetuses. Two principal sows that farrowed 65 and 67 days PI delivered 25 live piglets and three stillborn fetuses. The PRRS virus was isolated from two live piglets in one litter. In experiment 2, laparotomies were performed on five sows between 40 and 45 days of gestation and fetuses were inoculated in utero with either PRRS virus alone, PRRS virus plus a swine serum containing PRRS antibodies, or uninfected cell culture lysate. Three sows were euthanized on PI day 4 and two sows on PI day 11. Viral replication occurred in fetuses inoculated with virus alone and was enhanced in fetuses inoculated with virus plus antibody. No virus was isolated from control fetuses.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1992

Cytokine release from microglia: differential inhibition by pentoxifylline and dexamethasone.

Cytokines have been implicated in the pathogenesis of gram-negative bacterial meningitis. The effects of pentoxifylline and dexamethasone on the release of tumor necrosis factor (TNF), interleukin (IL)-1, and IL-6 from primary murine microglial cell cultures were explored using bioassays. When added concomitantly with lipopolysaccharide, pentoxifylline blocked the release of TNF and IL-1 but not IL-6, while dexamethasone inhibited the release of TNF and IL-6. After a 2-h exposure of microglia to lipopolysaccharide, pentoxifylline but not dexamethasone still inhibited the release of TNF. Release of TNF was enhanced 20-fold by priming of the microglia with interferon-gamma; only pentoxifylline blocked the priming effect of interferon-gamma on TNF release. These results demonstrate that pentoxifylline and dexamethasone differentially regulate the release of cytokines in microglial cell cultures and provide potential insight into their role in the treatment of gram-negative bacterial meningitis.

Animals↗

PubMed1992

Restriction of porcine parvovirus replication in nonpermissive cells.

Swine testicle (ST) cells and Madin-Darby canine kidney (MDCK) cells differ in their ability to support replication of porcine parvovirus (PPV). Viral replication events in ST cells, a permissive cell type, and MDCK cells, a nonpermissive cell type, were compared in an attempt to elucidate putative mechanisms of restrictive virus replication. Radiolabeled PPV bound to the cell surface of both cell types equally well and the binding was shown to be PPV specific, indicating that the restriction was not at the cell surface level. In contrast, profound differences in intracellular events in PPV replication were observed between these two cell types. Synthesis of viral DNA was limited in MDCK cells in that the percentage of cells with replicative-form DNA as determined by strand-specific probe in situ hybridization was approximately 100-fold lower in MDCK cells than in ST cells at the same multiplicity of infection. Northern (RNA) blot analysis, using oligonucleotide probes derived from both structural and nonstructural protein-coding regions of the PPV genome, revealed four PPV mRNA transcripts from infected ST cells. Comparatively, RNA species from the structural protein coding region were actively transcribed in MDCK cells, but synthesis of RNA species from the nonstructural protein coding region was negligible. Immunoprecipitation of viral polypeptides revealed the three characteristic structural polypeptides, VP1, VP2, and VP3, along with the nonstructural polypeptide, NS-1 from ST cells. In contrast, neither viral structural or nonstructural polypeptides nor progeny virions were produced from MDCK cells. The data suggest that mechanisms controlling permissiveness of cells to PPV infection are associated with the level of viral DNA replication, RNA transcription, and viral antigen expression but not absorption to the cell surface.

Animals↗

PubMed1991

Polymerase chain reaction (PCR) amplification for the detection of porcine parvovirus.

A polymerase chain reaction (PCR) amplification method was developed and evaluated to detect porcine parvovirus (PPV). A pair of 20-base primers and an oligonucleotide probe were derived from the DNA sequences common to two isolates of PPV, NADL-8 and NADL-2. The primers flanked 118-bp nucleotides within the region coding for the major structural protein VP2. After DNA amplification of PPV replicative form (RF), a 158-bp fragment was detected in agarose gels. This amplified fragment was shown to be specific for PPV DNA after Southern transfer and hybridization to a 20-base internal probe. The amplified fragment also contained a single EcoRI cleavage site. Various conditions, such as number of cycles and annealing temperature, were examined to optimize the conditions for detecting viral DNAs from infected cell cultures and swine fetal tissues. Four different isolates of PPV, NADL-8, NADL-2, KBSH and Kresse, and two other viruses, canine parvovirus (CPV) and pseudorabies virus (PRV), were included to determine specificity of amplification. Slot blot hybridization with a radiolabeled probe was used to evaluate the sensitivity of PCR amplification. The optimized protocol was specific for PPV detecting equally all four strains of PPV, but failing to amplify CPV or PRV sequences. The PCR method could detect at least 100 fg of viral replicative form (RF) DNA or the equivalent of 1 PFU of infectious virus. The applications of this method include routine detection of PPV in clinical samples and as a contaminant in mammalian cell lines.

Animals↗

PubMed1990

Structure of 1,4-cubanediyldiammonium bis(trinitromethanide).

C8H12N2+.2CN3O6-, Mr = 436.27, monoclinic, C2/m, a = 14.298 (2), b = 8.408 (1), c = 7.354 (2) A, beta = 103.42 (2) degrees, V = 859.9 (5) A3, Z = 2, Dx = 1.68 g cm-3, lambda(Mo K alpha) = 0.71069 A (graphite monochromator), mu = 1.66 cm-1, F(000) = 448, T = 293 K, final R = 0.039 for 672 reflections with I greater than or equal to 3 sigma(I). The cubanediyldiammonium cation and trinitromethanide anion have 2/m and m crystallographic symmetries, respectively. The cation is linked to six trinitromethanide anions, three at each end, by a total of 12 N--H...O hydrogen bonds through the six cation H atoms. The cubane cage has a local threefold axis of symmetry along the long axis (N1...N1) of the cation and undergoes a large librational motion, 17 degrees r.m.s. amplitude, about this axis.

Bridged-Ring Compounds↗

PubMed1990

Establishment of transformed swine fibroblast cell lines using SV40 large T antigen.

Swine testicle cell lines were established by transformation of primary swine testicle (PST) cells with an SV40 plasmid (pSV3-neo), which contains genes conferring resistance to neomycin and expressing SV40 large T antigen. Plasmid DNA was transfected into PST cells using a lipofection system. Two related plasmids, pSV2-neo and pSV5-neo, failed to induce transformed cells. Cells transformed with pSV3-neo formed single colonies that were resistant to the antibiotic, G418, and expressed large T antigen. Upon two cycles of cloning by endpoint dilution method, three transformed clones, designated transformed swine testicle (tST)-3, tST-14 and tST-18, were selected and characterized in regards to cell replication and susceptibility to swine viruses. The resultant clones were compared with a counterpart non-transformed ST cell line (ATCC-ST). The three tST cell lines showed longer or the same doubling times and higher saturation densities compared to ATCC-ST cells. These cells were free from a range of adventitious agents and supported the replication of porcine parvovirus (PPV), pseudorabies virus (PRV) and transmissible gastroenteritis virus (TGEV), comparable to ATCC-ST cells. All three cell lines have been maintained in continuous cultures for over 60 passages with no changes in growth characteristics. These findings indicate that lipofection with pSV3-neo is an efficient means for the introduction of exogenous DNA into porcine cells and for establishment of transformed immortalized cell lines.

Animals↗