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Biomedical subjects

C Hong

Publications and source records attributed to C Hong.

At least 73 records · Page 4Linked to original sources

Characterisation of fast, slow and cardiac muscle tropomyosins from salmonid fish.

Tropomyosin (TM) has been isolated from the cardiac muscle, and fast and slow trunk (myotomal) muscles of the mature salmonid fish Atlantic salmon (Salmo salar) and rainbow trout (Salmo gairdneri). When examined electrophoretically, isoforms of TM were detected which were specific, and exclusive, to each type of muscle. Cardiac and fast muscles contained single and distinct isoforms, while slow muscle contained two distinct isoforms, closely related in terms of apparent M(r), and pI. There was no detectable difference between the same TM type from either salmon or trout. On a variety of gel systems, the cardiac and slow isoforms migrated in close proximity to each other and to rabbit alpha-TM. The fast isoform comigrated with rabbit beta-TM. In developing salmon fry, a more acidic (unphosphorylated) variant of TM was present in addition to, and of similar M(r) to, the fast adult isoform. This TM declined in steady-state level during maturation and was virtually undetected in adult muscle. All of the isolated TMs contained little or no covalently bound phosphate and were blocked at the N-terminus. The amino acids released by carboxypeptidase A, when ordered to give maximal similarity to other muscle TMs, were consistent with the following sequences: fast (LDNALNDMTSI) and cardiac (LDHALNDMTSL). The C-terminal region of the slow TM contained His but was heterogeneous. In viscosity measurements, performed as a function of increasing protein concentration, at low ionic strength (t = 5 degrees C, pH 7.00), fast TM exhibited the highest relative viscosity values. Lower and equivalent levels of polymerisation occurred with the cardiac and slow TMs. Polymerisation of all three isoforms was temperature-dependent, with cardiac TM being least sensitive and fast TM being most sensitive. Determination of the complete coding sequence of adult fast TM confirmed the findings of the carboxypeptidase analysis, but the remainder of the sequence more closely resembled alpha-type TMs than beta-type TMs. Overall, salmon fast TM contains 20 (mostly conservative) substitutions compared to rabbit striated muscle alpha-TM and 40 (mostly conservative) substitutions compared to rabbit striated muscle beta-TM. This demonstrates that electrophoretic mobility is not, in all instances, a suitable method to assess the isomorphic nature of striated muscle TMs.

Amino Acid Sequence↗

Variations in the vascular pedicle of the rat gracilis muscle flap.

The rat gracilis muscle is a reliable model for microsurgical research. Exploration of 364 inguinal regions and preparation of 80 free gracilis muscle flaps in Sprague-Dawley and Lewis rats revealed six distinct origins of the dominant arterial pedicle and four variations of the venous pedicle. Popliteal, external iliac, and truncus pudendoepigastricus associations are described for the first time in this study. The total incidence of atypical variation of arterial pedicle origin is 20.7 percent. The most common origin of muscular branch arteries are the superficial epigastric artery (53.3 percent), femoral artery (22.0 percent), popliteal artery (14.6 percent), and the external iliac arteries (5.5 percent). Muscular branch veins drain most frequently into the femoral vein (79.1 percent). Gracilis muscle flaps prepared from rats weighing 329 +/- 45 g weighed an average of 0.64 +/- 0.13 g, and their volume averaged 0.87 +/- 0.12 ml. Muscular branch artery and vein diameters (external) averaged 0.21 +/- 0.05 mm and 0.54 +/- 0.06 mm, respectively. This new anatomic information is essential for reproducible microsurgery research using the rat gracilis muscle.

Animals↗

Human cytomegalovirus (CMV) DNA in plasma reflects quantity of CMV DNA present in leukocytes.

A quantitative DNA amplification assay for human cytomegalovirus (CMV) DNA has been used to evaluate the relationship between quantities of CMV DNA in plasma and those in infected leukocytes (WBC) from human immunodeficiency virus-infected patients. The target sequence for DNA amplification was a region of the immediate-early 1 gene of CMV. The quantitation assay uses an internal control that is coamplified with each patient sample DNA and contains a sequence for detection by colorimetric hybridization with the same bases, but in different order than in the CMV immediate-early 1 region used for hybridization of amplified patient sample DNA. Results showed that patients with CMV disease had more CMV DNA in both WBC and plasma than those without disease. However, in this study, copy numbers of CMV DNA in WBC were higher than those in plasma. The gB and gH variants were the same in plasma and WBC.

Base Sequence↗

Immunomodulating effects of synchronised plasmapheresis and intravenous bolus cyclophosphamide in systemic lupus erythematosus.

Recent studies have suggested that synchronised plasmapheresis and intravenous pulse cyclophosphamide therapy reduce disease activity in SLE patients. The aim of the present study was to examine the immunomodulating effects of this therapy and compare it with changes seen with cyclophosphamide alone. Four patients with active SLE were studied. Two were treated with synchronised therapy and two received cyclophosphamide only for up to 26 weeks. Disease activity was measured by the SLE disease activity index (SLEDAI). Immunological studies were performed immediately prior to each treatment. Patients in both treatment groups improved as reflected by a fall in mean SLEDAI scores: synchronised therapy 33.5 to 11; cyclophosphamide only 13.5 to 4.5. Following synchonised therapy only there was a prompt and sustained increase in the mean percentage of CD8+ cells (20.8 to 54.8) which resulted in a fall in the CD4:CD8 ratio (1.95 to 0.62). With both treatment modalities there was a fall in the proportion of CD20+ cells (B lymphocytes) (synchronised therapy 10.5 to 3.2; cyclophosphamide only 5.6 to 2.2). However, only synchronised therapy resulted in a fall in the in vitro production of immunoglobulins which was unchanged or increased following cyclophosphamide alone. These results suggest that although both treatment modalities are efficacious in the treatment of active SLE they produce different immunomodulatory effects. Thus, both therapies reduce the number of circulating B lymphocytes whereas synchronised therapy also modifies cellular immunity by promoting the emergence of a phenotypic suppressor T lymphocyte population.

Adjuvants, Immunologic↗

Probability of global indices in low tension glaucoma.

To know the pattern of visual field (VF) defect of low tension glaucoma (LTG), its diffuse and localized VF defects were compared using the significance values (p-value) of mean deviation (MD) and corrected pattern standard deviation (CPSD), which are calculated with STATPAC in Humphrey Visual Field Analyser. Sixteen eyes of LTG were enrolled and 34 eyes of primary open angle glaucoma (POAG) were used as controls. The degree of VF defects in LTG and POAG was relatively mild with the p-value of MD equal to or greater than 1%. Neither mean MD nor mean CPSD of LTG was significantly different from each of POAG (p > 0.8 and p > 0.2, respectively). Comparing the p-values of MD and CPSD, many patients showed more significant p-value of MD in LTG and POAG (62.5% and 61.8%, respectively). However, relatively more severe defect of CPSD tended to occur more frequently in LTG than in POAG (25% and 5.9%, respectively, chi 2 = 4.964, p < 0.09). The relation between p-values of MD and CPSD was not significantly influenced by the intraocular pressure, MD or vertical cup to disc ratio in either LTG or POAG.

Adolescent↗

Requirement of beta 2 glycoprotein I as cofactor in the binding for IgM and IgA anticardiolipin antibodies.

OBJECTIVE: To determine if IgM and IgA anticardiolipin (aCL) antibodies require beta 2 glycoprotein I (beta 2-GPI) as a cofactor for antibody binding. METHODS: Sera were selected from 7 patients with systemic lupus erythematosus (SLE), 6 of whom had high IgM and 6 high IgA aCL antibody binding. Control sera were obtained from 2 healthy individuals with no aCL antibodies. Serum proteins were initially separated by sepharose CL6B get filtration chromatography, and IgM was further purified by affinity chromatography with mannan binding protein. IgA was isolated from CL6B filtrate by jacalin lectin affinity chromatography. Levels of beta 2-GPI in the immunoglobulin preparations were determined by antigen capture ELISA: Anticardiolipin antibody binding IgM and IgA was examined by ELISA with and without the addition of beta 2-GPI (10 micrograms/ml) or 4% normal human serum and expressed in optical density units (OD). RESULTS: beta 2-GPI was required as a cofactor for IgM aCL antibody binding in 4 to 6 patients with SLE. In these, antibody binding to cardiolipin increased from (mean +/- SEM) 0.10 +/- 0.01 TO 1.06 +/- 0.22 (p = 0.005) with the addition of beta 2-GPI. For IgA, 5 of 6 patients with SLE demonstrated a requirement of beta 2-GPI as a cofactor. Antibody binding increased from 0.27 +/- 0.05 to 1.77 +/- 0.35 (p = 0.003) with the addition of beta 2-GPI. CONCLUSION: beta 2-GPI is required as a cofactor for IgM and IgA aCL antibody binding.

Antibodies, Anticardiolipin↗

A new photopolymerizable blood vessel glue that seals human vessel anastomoses without augmenting thrombogenicity.

A new nonbiologic photopolymerizable glue, polyethyleneglycol 400 diacrylate, was studied with respect to its mechanical and biochemical interaction with human blood vessels. Using the human placental artery model, we tested the ability of polyethyleneglycol 400 diacrylate to prevent leakage of blood at the site of vascular anastomoses, which are made porous by the presence of tissue gaps and suture puncture sites. Fibrin glue is known to augment local vessel thrombogenicity through the presence of the coagulation enzyme thrombin. We tested the effect of externally applied polyethyleneglycol 400 diacrylate (which does not contain thrombin) on luminal thrombin activity and platelet deposition from flowing human blood. At a shear rate of 312 per second and a transmural pressure of 80 cm H2O, the leakage rate of saline from human placental artery anastomoses was 1.0 +/- 1.2 ml/min (n = 8). When the same anastomoses were then glued, 7 of 8 of the anastomoses leaked less than 0.05 ml/min (p < 0.05). Platelet deposition to human vessels was not influenced by the external application of polyethyleneglycol 400 diacrylate either on intact vessels (no polyethyleneglycol 400 diacrylate, 0.51 +/- 0.28 x 10(6) platelets/cm2; with polyethyleneglycol 400 diacrylate, 0.47 +/- 0.26 x 10(6) platelets/cm2; n = 7) or at anastomoses (no polyethyleneglycol 400 diacrylate, 0.69 +/- 0.36 x 10(6) platelets/cm2; with polyethyleneglycol 400 diacrylate, 0.53 +/- 0.33 x 10(6) platelets/cm2; n = 8), p > 0.05.(ABSTRACT TRUNCATED AT 250 WORDS)

Anastomosis, Surgical↗

Detection of anthelmintic resistant nematodes in sheep in southern England by a faecal egg count reduction test.

On five farms identified by in vitro tests to have benzimidazole resistance, and on one farm with apparent anthelmintic failure, faecal egg count reduction tests were performed using benzimidazoles, levamisole and ivermectin. With benzimidazoles efficacy was less than 70 per cent on all farms, confirming the validity of the in vitro tests to detect benzimidazole resistance. On two farms levamisole had an efficacy of less than 90 per cent but laboratory tests failed to demonstrate levamisole resistance. Ivermectin reduced egg counts by 100 per cent on all farms.

Animals↗

Posttranscriptional modification of myosin heavy-chain gene expression in the hypertrophied rat myocardium.

Hypertrophy of the myocardium in response to pressure or volume overload elicits a change in myofibrillar protein content as a result of changes in both transcriptional and translational regulation of gene expression. Hemodynamic overload caused by aortic constriction produced changes in the expression of the two isoforms of myosin heavy chain (MHC) with a 319% increase in beta-MHC mRNA and a 54% decrease in alpha-MHC mRNA (P < 0.01). Cardiac unloading as a result of heterotopic transplantation resulted in a decrease in cardiac mass and a similar shift in MHC isoform expression. In this study. We investigated cardiac gene transcription to understand how different hemodynamic stimuli produce similar cardiac phenotypes. We studied the in vivo activity of the alpha-MHC promoter (-2564 to +421 bp of the transcriptional start site) by directly injecting a recombinant expression plasmid (pAM3LUC) into the ventricular tissue of coarctated animals as well as into the unloaded heterotopic transplanted heart. When expressed as a function of the activity of a constitutively active viral promoter (pSVCAT), pAM3LUC activities were 18.4 +/- 2.9, 24.6 +/- 2.6, and 25.0 +/- 4.5 (x10(4)) luciferase/chloramphenicol acetyltransferase units in the hypertrophied ventricles of 2-, 3-, and 7-day coarctated animals, respectively. These values were not statistically different from pAM3LUC activity in control hearts of sham operated animals even though alpha-MHC mRNA content was decreased by 54% in the hypertrophied myocardium. This disparity between transcriptional activity and mRNA content suggests that alpha-MHC expression in the hypertrophic ventricle is in part regulated by a posttranscriptional mechanism. In contrast, alpha-MHC promoter activity in the unloaded transplanted hearts decreased significantly by 37% compared to control working hearts and suggests that a transcriptional mechanism of regulation of the alpha-MHC gene may account for the phenotypic expression observed in the unloaded myocardium.

Animals↗

Isolation and characterization of tropomyosin from fish muscle.

A comprehensive survey of tropomyosin from various fish myotomal muscles is reported. The fish tropomyosins were blocked at the N-terminus and, as expected, were found to be of similar amino acid composition, alpha-helical content (> 90% at 10 degrees C) and molecular weight to other vertebrate striated muscle forms. The tropomyosins of salmonids and herring muscle were noticeably heterogeneous when assessed by 2D-PAGE. The distribution of isoforms was tissue-specific: slow muscle contained alpha-type tropomyosin while fast muscle contained beta-type tropomyosin. In other species (cod, haddock, wolf-fish and sharks) alpha-type tropomyosins were present in both kinds of muscle but beta-tropomyosin was absent.

Amino Acid Sequence↗

The effects of intraoperative mitomycin-C or 5-fluorouracil on glaucoma filtering surgery.

We compared the success rate of filtering surgery of a single 5-minute intraoperative application of mitomycin-C (MMC) or 5-fluorouracil (5-FU). Animal experiment and clinical study were done. In animal study, thirty pigmented rabbits (60 eyes) weighing 2.0 to 2.5 Kg were enrolled. We divided into 4 groups, such as BSS, 5-FU 50 mg/ml, MMC 0.2 mg/ml and MMC 0.4 mg/ml subconjunctival soaked group. Each group consisted of 15 eyes. In each group, 10 eyes was for examination of bleb survival and complications, 3 eyes for light microscopic examination and 2 eyes for electron microscopic examination. Bleb was survived 6.1 days (3 to 13 days) for BSS treated group, 16.3 days (9 to 23 days) for 5-FU 50mg/ml treated group, 32.7 days (17 to 55 days) in MMC 0.2 mg/ml treated group, and 64.4 days (49 to 84 days) in MMC 0.4 mg/ml treated group. Duration of bleb survival was significantly prolonged in 5-FU, MMC 0.2 mg/ml and MMC 0.4 mg/ml group respectively. In clinical study, fifty-five eyes of 40 patients were enrolled. 29 eyes of 21 patients were treated with MMC 0.2 mg/ml and 26 eyes of 19 patients were treated with 5-FU 50 mg/ml. The success rate was 89.7% in MMC 0.2 mg/ml treated group and 84.6% in 5-FU 50 mg/ml treated group at postoperative 3 months, and postoperative 6 months 89.7% in MMC 0.2 mg/ml treated group and 76.9% in 5-FU 50 mg/ml treated group. There was no statistical significance at postoperative 3 months (P > 0.05), but statistical significance at postoperative 6 months (P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Effects of adrenergic agonists on the growth and gene expression of the transplanted heart.

Cardiac growth occurs in response to changes in hemodynamic demand and results primarily from cellular hypertrophy without cellular hyperplasia. In addition to changes in cardiac work, various reports have demonstrated that adrenergic stimulation, specifically of the alpha 1 adrenergic receptor of cultured myocytes, can produce changes in cardiac-specific gene expression and increases in protein synthesis and cell growth. To study the effects of adrenergic stimulation in the absence of alterations in cardiac work we have used the model of the heterotopically transplanted heart, which is a spontaneously beating, vascularly perfused, and histologically normal heart that is hemodynamically unloaded. Seventy-two hours after transplantation, the hemodynamically unloaded transplanted heart had decreased in size by 20% when compared with the control host heart that was growing in situ. Treatment with either the alpha-adrenergic agonist phenylephrine (2.5 mg/kg/day) or the beta-adrenergic agonist isoproterenol (250 micrograms/kg/day) while increasing the spontaneous heart rate had no effect on the size of the transplanted heart. Simultaneous measurements demonstrated that both of these drugs produced a 10% increase in weight of the corresponding hemodynamically loaded host heart. We previously demonstrated that cardiac unloading caused a change in the expression of the myosin heavy chain (MHC) genes with a significant increase in the beta MHC isoform. In the present studies treatment with either isoproterenol or phenylephrine did not alter MHC gene expression either in the in situ host or transplanted hearts.(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenergic Agonists↗

Activity of morantel citrate against strains of benzimidazole-resistant nematodes of sheep in the United Kingdom.

The activity of morantel citrate (5.94 mg/kg base) was determined in laboratory tests against field isolates of benzimidazole-resistant nematodes. Its efficacies against adult and seven-day-old worms were 100 per cent and 100 per cent for Cooperia curticei, 95.1 per cent and 69.8 per cent for Haemonchus contortus and 100 per cent and 82 per cent for Ostertagia circumcincta. Morantel citrate was 100 per cent effective against benzimidazole-susceptible Nematodirus battus and Trichostrongylus vitrinus, and it reduced faecal egg counts by 97.9 per cent in sheep infected naturally with benzimidazole-resistant H contortus and O circumcincta.

Animals↗

Artificial microvascular graft thrombosis: the consequences of platelet membrane glycoprotein Ib inhibition and thrombin inhibition.

Early thrombosis of artificial microvascular grafts (AMG, grafts < or = 2 mm internal diameter) prevents their reliable clinical use. The present studies were undertaken to examine the effect of hirudin, a thrombin-specific inhibitor, and of the F(ab')2 fragment of PG-1, a monoclonal antibody (MoAb) directed against guinea pig platelet membrane glycoprotein Ib (GPIb), on AMG patency in an animal model. One-centimeter long segments of expanded polytetrafluoroethylene (ePTFE), 0.88 mm internal diameter, were serially implanted as interposition grafts in the guinea pig femoral arterial systems bilaterally. A control group was treated with 0.5 mL saline intravenously (IV) 30 minutes before limb 1 and limb 2 graft implantation. Three experimental groups were treated with 0.5 mL saline IV before limb 1 graft implantation as an animal control and with either 0.5 mL saline containing 1.25 mg/kg IV PG-1 F(ab')2, (which inhibits ristocetin-induced platelet agglutination and von Willebrand factor binding), hirudin 1 mg/kg IV, or a combination of both agents before limb 2 graft implantation. GPIb inhibition, thrombin inhibition, and the combination resulted in a significant prolongation of AMG patency (P < .005). Whereas thrombin inhibition with hirudin prolonged AMG patency similar to that observed with GPIb inhibition, the combination of GPIb and thrombin inhibition provided the overall longest prolongation of AMG patency. These results indicate that both platelet membrane GPIb and thrombin play a role in AMG thrombosis.

Adenosine Triphosphate↗

Antibodies to brain integral membrane proteins in systemic lupus erythematosus.

Wheat germ lectin affinity chromatography and temperature-induced phase separation with Triton X-114 were evaluated for the isolation of surface neuronal antigens from rat and human brain and from human neuroblastoma cell lines IMR-6 and SK-N-SH. Both techniques yielded surface proteins which were free of contamination by intracellular proteins but temperature-induced phase separation was technically less demanding and less expensive, required a shorter assay time and resulted in a superior quantity and quality of isolated proteins. Rat brain surface proteins were used for characterization of antineuronal antibody reactivity in sera from patients with systemic lupus erythematosus (SLE). Western blotting identified reactivity in 15 of 75 (20%) SLE sera compared to five of 95 (5%) normal controls (P 0.006). In rat brain the molecular weight of the individual proteins identified ranged from 59 kDa to 22 kDa. Six of these were also present in human brain and two were present in neuroblastoma cell lines. Absorption studies indicated that some of the antigenic proteins were either restricted to brain tissue or shared with other non-neuronal tissues. These techniques should facilitate the characterization of antineuronal antibody reactivities and lead to a clearer understanding of their role in the pathogenesis of autoimmune neurologic disease.

Adult↗

Persistent efficacy of doramectin against experimental nematode infections in calves.

Three studies were conducted involving cattle exposed to experimental nematode infections. These studies were designed to investigate the prophylactic activity of a single subcutaneous treatment of doramectin at 200 micrograms kg-1 body weight against infections of Ostertagia ostertagi, Cooperia oncophora and Dictyocaulus viviparus. For each study, parasite-naive calves were randomly allocated to either a treated or a matched control group. One group received doramectin and the other received doramectin and the other received either no treatment or an injection of saline at 1 ml per 50 kg body weight by the subcutaneous route. Thereafter, all calves received a daily oral challenge of infective larvae of the particular parasite species on test in each study. Challenge of each pair of treatment/control groups continued for periods of 14, 21 or 28 days. An interval of 14-21 days was then allowed to permit the parasites which had established to mature, after which all animals were slaughtered and their worm burdens determined using standard techniques. Geometric mean worm burdens were calculated from the log worm counts and used to estimate percentage efficacy. Accumulated burdens of C. oncophora in doramectin-treated cattle resulting from a daily challenge infection for 14 or 21 days were reduced by 99.2% and 90.7% respectively, in comparison with those of non-treated control animals. For D. viviparus, burdens were reduced by 100% and 99.9% after a 21 or 28 day challenge, respectively. The corresponding figures for O. ostertagi were 99.9% after a 21 day challenge and 93.7% after a 28 day challenge.

Analysis of Variance↗