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Biomedical subjects

T J Johnson

Publications and source records attributed to T J Johnson.

At least 37 records · Page 2Linked to original sources

Effects of jejunoileal autotransplantation on gastrointestinal regulatory peptides.

Plasma gastrointestinal hormones were measured before and during feeding in eight dogs, more than one year after total autotransplant of the entire jejunoileum, and in controls. At sacrifice, tissues were taken from the transplanted segment and intact bowel for measurement of hormone and enteric neuropeptide content. Gastrin levels were reduced in autotransplanted dogs (fasting 63% of control, incremental response 67% of control, both P < 0.05), reflecting the loss of acid inhibitory reflexes. Secretin and cholecystokinin responses were identical between the two groups. Postprandial levels of gastric inhibitory peptide (incremental response 175% of control, P < 0.005), insulin, and peptide YY (158% of control, P < 0.05) were elevated following denervation, the former suggesting more rapid gastric emptying while the latter may reflect malabsorption. The neurotensin meal response was obtunded by denervation (incremental response 43% of control, P < 0.05), providing evidence for a neural pathway for its release. Pancreatic polypeptide responses were identical between the groups, suggesting intact pancreatic innervation. Abnormal hormone secretion may contribute to the impaired fed motor responses seen following extrinsic denervation of the small bowel. In contrast, the neuropeptide content of the autotransplanted small intestine is normal, suggesting that extrinsic denervation has no long-term effects on peptide content of the enteric nervous system.

Animals↗

Bacterial translocation in the portal-hypertensive rat: studies in basal conditions and on exposure to hemorrhagic shock.

BACKGROUND: Portal hypertension is associated with altered intestinal motor and mucosal function. The aim of this study was to determine whether portal hypertension, per se, or in association with acute hemorrhagic shock, predisposes to the translocation of bacteria across the intestine. METHODS: Translocation to both mesenteric lymph nodes and blood was compared in three groups of rats: portal-hypertensive (single-stage calibrated stenosis of portal vein), sham-operated, and unoperated controls. Half of the animals in each group were exposed to hemorrhagic shock. RESULTS: In the basal state, translocation to both mesenteric lymph nodes (portal hypertension vs. sham vs. controls, 411.5 +/- 119 vs. 151.1 +/- 42.6 vs. 18.1 +/- 12.6 colony-forming units [CFU]/g; P < 0.05) and blood (portal hypertension vs. sham vs. controls, 100% vs. 30% vs. 0% positive blood cultures; P < 0.05) was significantly increased in the portal-hypertensive animals. Furthermore, translocation was strikingly increased in these animals after hemorrhagic shock (mesenteric lymph node cultures, portal hypertension vs. portal hypertension with shock, 411 +/- 119 vs. 1018.2 +/- 372.2 CFU/g; P < 0.05). CONCLUSIONS: Portal hypertension promotes bacterial translocation, especially in relationship to acute hemorrhage. These findings may, in part, explain the susceptibility of patients with liver disease to sepsis of enteric origin.

Animals↗

Chromosomal influence on hybrid cell proliferation.

When normal cells and cancer cells (usually from the same species) are experimentally fused the resultant hybrid cells show loss of the tumorigenic phenotype. To examine the phenotypic phenomenon of growth suppression in hybrid cells in vitro, we examined the doubling times of somatic cell hybrids which contained single or multiple chromosomes derived from another species (inter-species hybrids). In all of the hybrid lines examined, the presence of transferred chromosomes prolonged the cell population doubling times in proportion to the number of such chromosomes found in the hybrid lines. These findings provide additional evidence to support the hypothesis that increasing the genetic burden of cells may reduce the division potential of cells cultured in vitro.

Animals↗

Visualization of Trichoderma reesei Cellobiohydrolase I and Endoglucanase I on Aspen Cellulose by Using Monoclonal Antibody-Colloidal Gold Conjugates.

Monoclonal antibodies (MAbs) specific for cellobiohydrolase I (CBH I) and endoglucanase I (EG I) were conjugated to 10- and 15-nm colloidal gold particles, respectively. The binding of CBH I and EG I was visualized by utilizing the MAb-colloidal gold probes. The visualization procedure involved immobilization of cellulose microfibrils on copper electron microscopy grids, incubation of the cellulose-coated grids with cellulase(s), binding of MAb-colloidal gold conjugates to cellulase(s), and visualization via transmission electron microscopy. CBH I was seen bound to apparent crystalline cellulose as well as apparent amorphous cellulose. EG I was seen bound extensively to apparent amorphous cellulose with minimal binding to crystalline cellulose.

Journal Article↗

Repricing plan yields realistic revenue enhancement.

Actual reimbursement plays a key role in hospital rate restructuring. Moving away from a policy of rate setting "across-the-board" and toward modifying charges that positively affect a hospital's bottom line can more effectively generate net operating revenue, increase cash flow, and reduce increases in contractual allowances. A participatory approach between senior executives, managers, and physicians is working at several hospitals.

Accounting↗

Quantitative determination of selected compounds in a Kentucky 1R4F reference cigarette smoke by multidimensional gas chromatography and selected ion monitoring-mass spectrometry.

Eight compounds from a Kentucky 1R4F reference cigarette smoke condensate have been determined by selected ion monitoring-mass spectrometry (SIM-MS) to confirm the validity of multidimensional gas chromatography (MDGC) as a quantitative tool in complex mixture analyses. Four electrostatically precipitated smoke condensate samples of 100 cigarettes each are dissolved individually in 25 mL of 2-propanol. The 2-propanol contains two methyl esters (C8 and C14) and seven deuterium-labeled compounds used as internal standards (IS). Analysis of the compounds of interest, pyridine; acetamide; acrylamide; phenol; o-, m-, and p-cresol; and quinoline, is accomplished by using two heartcuts. Heartcut times of the MDGC analysis are selected such that at least one IS is transferred with each group of compounds being analyzed. This study shows that the MDGC technique previously developed and described can be used for quantitative analyses. A comparison is made between the two types of internal standards. The results obtained for both types of internal standards agree within 20% of each other, on the average, with higher standard deviations for approximately 60% of the compounds where methyl esters are used as internal standards.

Chromatography, Gas↗

Potential for fecundin to influence the reproductive performance of merino ewes in Western Australia.

Four experiments were conducted at different locations in Western Australia to evaluate the effectiveness of immunizing young (maiden, 1 1/2 year old) and adult Merino ewes with Fecundin to improve their reproductive performances. The ovulation rates of immunized maiden ewes was increased (0.06-0.30) above that of untreated ewes in both experiments 1 and 2. However, there were no significant improvements in the marking percentages for the immunized ewes with the differences between the untreated and immunized ewes ranging from -16.4 to 5.8%. In comparison with untreated ewes immunization 6 and 2 weeks before the start of joining depressed ewe fertility by 26.1% whereas immunization 8 and 4 weeks before the start of joining did not significantly affect fertility. The availability of oat grain ad libitum prior to parturition and during early lactation did not improve the survival of lambs born to immunized ewes. In experiments 3 and 4 immunization of adult Merino ewes increased their ovulation rates (0.41-0.63) above untreated controls and tended to increase the proportion of pregnant ewes which had multiple pregnancies (from -2.3 to 34.2%). The responses at the end of lambing were variable (from -19.8 to 37.5% lambs marked) with high lamb mortalities occurring in some experiments. There was no adverse effect on the reproductive performances following consecutive annual immunizations over 3 years and the absence of treatment for 1 year did not prevent a response in the following year.

Aging↗

Postreplication labeling of E-leaflet molecules: membrane immunoglobulins localized in sectioned, labeled replicas examined by TEM and HVEM.

Conventional freeze-fracture techniques were combined with immunogold labeling and with plastic embedding and sectioning to analyze the distribution of membrane immunoglobulins (mIgs) and their associated intramembrane particles (IMPs) in E-face replicas of murine B-lymphocyte plasma membranes. Immunogold labels were applied to cells after the process of freeze-fracture and replication. Conventional stereoscopic transmission electron microscopic examination of sectioned, labeled replicas (SLRs) revealed that the gold-labeled mIgs were bound to and localized on the outer leaflets of split and replicated membranes. The gold labels were attached to the external determinants of the mIg molecules, which were retained beneath and contiguous with the replicated E-faces. The mIgs were also localized on the external surface of unreplicated microvilli. In addition, thick sections examined by high-voltage transmission electron microscopy (HVEM) revealed large expanses of replica with well-resolved IMPs. mIgs colocalized with small-diameter (less than 60 A) IMPs in E-face replicas of B-lymphocytes whose mIgs were patched by anti-immunoglobulin. Thus, postreplication E-surface labeling of split and replicated membranes is a high-resolution technique that is suitable for the study of membrane protein distribution in E-face replicas and contiguous nonreplicated tissue.

Animals↗

Glutaraldehyde fixation chemistry: oxygen-consuming reactions.

In tissue fixed with glutaraldehyde, dissolved O2 is rapidly consumed by two processes: residual respiration and glutaraldehyde-induced chemical uptake. The nature of the chemistry which consumes O2 during tissue fixation was investigated by studying model reactions of glutaraldehyde with amines and with homogenized tissue suspensions. The addition of glutaraldehyde to solutions of most primary amines and ammonia stimulated rapid O2 consumption. The reaction of glutaraldehyde with primary amines (e.g., 25 mM ethanolamine, glycine, or methylamine) consumed 50% of the dissolved O2 in 15 to 20 s at 37 degrees C. The initial rate of O2 uptake followed second-order kinetics with respect to the primary amine concentration. The total amount of O2 consumed was sufficient to account for the stoichiometric conversion of the primary amines to pyridines. These data are consistent with the synthesis of pyridine derivatives from glutaraldehyde-amine precursors in which the last step is an irreversible oxidation of dihydropyridines to pyridines. The addition of glutaraldehyde to homogenized muscle suspensions, in which respiration was chemically inhibited, significantly increased the rate of O2 uptake. Thus, in tissue O2 is rapidly depleted both by respiration and the chemical demands of the glutaraldehyde-amine reactions during the cross-linking process. Since these experiments were done under conditions commonly used for tissue fixation, hypoxia should be assumed to exist in biological preparations fixed with glutaraldehyde.

Aldehydes↗

Labelled-replica techniques: post-shadow labelling of intramembrane particles in freeze-fracture replicas.

Three methods are described for direct post-fracture, post-shadow labelling of individual classes of intramembrane particles (IMPs) in freeze-fracture replicas of biological membranes. The P-face IMPs corresponding to the acetylcholine receptor complexes (AChRs) of vertebrate neuroeffector junctions are identified by post-replication labelling with ferritin-antibody complexes and with neurotoxin-biotin-avidin-colloidal gold affinity ligands. (The freeze-etch nomenclature of Branton et al., 1975, is used in this report.) These post-shadow labelling techniques resemble conventional en bloc labelling techniques except that the labelling reagents must penetrate a thin but discontinuous layer of platinum superimposed on the molecules of interest. In the 'sectioned labelled-replica technique', the replicated and labelled tissues are stained, embedded in plastic and sectioned parallel to the replica-tissue interfaces. In the direct 'labelled-replica techniques', the replicated and labelled samples are freeze-dried or critical point dried, the labelled surfaces are stabilized by carbon coating, and the underlying tissues are dissolved, allowing the labelled-replicas to be examined as conventional freeze-fracture replicas. The unshadowed side of each AChR IMP is shown to retain sufficient biochemical information to permit both immunospecific and neurotoxin specific labelling despite formaldehyde fixation, freezing, fracturing, platinum shadowing, and thawing in aqueous media. A new mixed ferricyanide-osmium staining method reveals electron opaque structures spanning the membrane bilayer in the same size, number and distribution as the labelled IMPs. These experiments demonstrate the feasibility of identifying individual IMPs in freeze-fracture replicas and may allow the identification of specific membrane lesions in human disease.

Animals↗