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Biomedical subjects

C M Johnson

Publications and source records attributed to C M Johnson.

At least 127 records · Page 7Linked to original sources

Soluble E-selectin is increased in inflammatory synovial fluid.

OBJECTIVE: To investigate the hypothesis that soluble E-selectin (sE-selectin) may be detected in synovial fluid (SF) and play a role in inflammatory arthritis. METHODS: We used a sandwich ELISA to measure sE-selectin in the SF of 58 patients with rheumatoid arthritis (RA), 9 with psoriatic arthritis (PsA), 30 with osteoarthritis (OA), 13 with gout, and 9 with calcium pyrophosphate dihydrate crystal deposition disease (CPPD). RESULTS: SF sE-selectin values in RA (mean 1.49 ng/ml, 0.18-3.90) and PsA (mean 1.36 ng/ml, 0.88-2.31) were significantly higher than those with OA (mean 0.83 ng/ml, 0.00-1.83), gout (mean 1.04 ng/ml, 0.11-3.42), or CPPD (mean 0.80 ng/ml, 0.20-1.47). Elevated SF sE-selectin was associated with elevated serum sE-selectin, erythrocyte sedimentation rate, and SF white blood cell count. CONCLUSION: Our findings suggest that endothelial cell activation and E-selectin may contribute to the development of inflammatory processes.

Adult↗

Feline lymphoid tissues engrafted into scid mice maintain morphologic structure and produce feline immunoglobulin.

To adapt the feline immune system to a small laboratory animal host, severe combined immunodeficient (scid) mice were engrafted with neonatal feline lymphoid tissues, including lymph node, thymus, spleen, and bone marrow. Lymph node and thymus tissue were implanted subcutaneously within the mammary fat pad, and a single-cell suspension of spleen, thymus, and bone marrow was inoculated intraperitoneally (IP). Seven groups of mice (three mice per group) were engrafted on day 0, and members of one group were euthanatized weekly from 2 to 8 weeks after engraftment. For each mouse, graft morphology was evaluated by light microscopy, feline DNA was detected in peripheral blood by polymerase chain reaction (PCR) amplification of feline-specific DNA sequences, and serum IgG concentration was measured by ELISA. Ten of 13 feline grafts evaluated histologically between 3 and 8 weeks after engraftment contained large focal aggregates of lymphocytes bordered by plasma cells. Of 14 thymus grafts evaluated histologically during the same period, 5 were characterized by dense accumulations of small lymphocytes surrounding thymic epithelial cells. Two of these thymus grafts were indistinguishable from age-matched feline thymus. At 2 weeks after engraftment, feline lymph node and thymus contained extensive central necrosis bordered by a narrow zone of lymphocytes and small-caliber blood vessels.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Effect of cavity-creating mutations in the hydrophobic core of chymotrypsin inhibitor 2.

Hydrophobic residues in the core of a truncated form of chymotrypsin inhibitor 2 (CI2) have been mutated in order to measure their contribution to the stability of the protein. The free energy of unfolding of wild-type and mutants was measured by both guanidinium chloride-induced denaturation and differential scanning calorimetry. The two methods give results for the changes in free energy on mutation that agree to within 1% or 2%. The average change in the free energy of unfolding (+/- standard deviation) for an Ile-->Val mutation is 1.2 +/- 0.1 kcal mol-1, for a Val-->Ala mutation 3.4 +/- 1.5 kcal mol-1, and for either an Ile-->Ala or a Leu-->Ala mutation 3.6 +/- 0.6 kcal mol-1. This gives an average change in the free energy of unfolding for deleting one methylene group of 1.3 +/- 0.5 kcal mol-1. Two significant correlations were found between the change in the free energy of unfolding between wild-type and mutant, delta delta GU-F, and the environment of the mutated residue in the protein. The first is between delta delta GU-F and the difference in side-chain solvent-accessible area buried between wild-type and mutant (correlation coefficient = 0.81, 10 points). The second and slightly better correlation was found between delta delta GU-F and N, the number of methyl/methylene groups within a 6-A radius of the hydrophobic group deleted (correlation coefficient = 0.84, 10 points). The latter correlation is very similar to that found previously for barnase, suggesting that this relationship is general and applies to the hydrophobic cores of other globular proteins. The combined data for C12 and barnase clearly show a better correlation with N (correlation coefficient = 0.87, 30 points) than with the change in the solvent-accessible surface area (correlation coefficient = 0.82, 30 points). This indicates that the packing density around a particular residue is important in determining the contribution the residue makes to protein stability. In one case, Ile-->Val76, a mutation which deletes the C delta 1 methyl group of a buried side chain, a surprising result was obtained. This mutant was found to be more stable than wild-type by 0.2 +/- 0.1 kcal mol-1. We have solved and analyzed the crystal structure of this mutant and find that there are small movements of side chains in the core, the largest of which, 0.7 A, is a movement of the side chain that has been mutated.(ABSTRACT TRUNCATED AT 400 WORDS)

Amino Acid Sequence↗

Skin excision revision rhinoplasty.

A technique of skin excision revision rhinoplasty is described for application in patients undergoing rhinoplasty. This group of patients includes those with soft-tissue polly beak deformities and a variety of other difficult post-rhinoplasty deformities. Horizontal nasal dorsal skin excision affects tip elevation and eliminates supratip fullness, while vertical midline nasal dorsal skin excision enhances tip definition, decreases dorsal height, and eliminates supratip fullness. We describe several patients who underwent either horizontal or vertical nasal dorsal skin excision for a variety of cosmetic deformities following conventional closed and open rhinoplasty. The techniques presented herein are meant to be added to the list of techniques available to the revision rhinoplasty surgeon rather than to replace existing techniques.

Adult↗

Transport of the critically ill child.

During the past several decades, the transport of critically ill patients to and between hospitals has gradually improved. The major indications that necessitate emergency transport for adult patients are trauma and acute cardiac disease, and the establishment of transport teams trained in the care of these conditions has improved the outcome of adult patients. In critically ill children, the indications for emergency transport differ from those in adults; pediatric patients are most likely to be transported for respiratory and neurologic emergencies. The outcome for critically ill children is optimal when a dedicated pediatric transport team functions in the setting of a regional critical-care outreach program. Such a program emphasizes stabilization of the child's condition at the local hospital, followed by transport to a pediatric intensive-care facility by a specially trained pediatric transport team. In this review, we present an overview of the principles and operating procedures of such pediatric transport teams.

Child↗

External cephalic version at term.

The case notes of women with singleton term (37 weeks' gestation and beyond) breech presentation and delivery were retrospectively reviewed. Thirty-two of the 72 women in the study group had attempted external cephalic version at term, with a success rate of 53% (17 women). The Caesarean section rate was significantly lower in the group which had attempted ECV compared to the group which did not.

Breech Presentation↗

In vivo lymphocyte tropism of feline immunodeficiency virus.

Feline immunodeficiency virus (FIV) infection in the cat is similar to human immunodeficiency virus type 1 infection in causing a selective reduction in CD4+ cell numbers, leading to inversion of the CD4+/CD8+ ratio. To determine whether FIV, similar to human immunodeficiency virus type 1, has a tropism for CD4+ cells, we examined the in vitro and in vivo susceptibilities of feline lymphocyte subpopulations to FIV infection. Infection of interleukin-2-dependent CD4+ or CD8+ lymphocyte cultures with the NCSU1 isolate of FIV (FIV-NCSU1) resulted in syncytium formation, cell death, and Mg(2+)-dependent reverse transcriptase (RT) activity in both cases. Monoclonal antibodies to feline lymphocyte subsets were used to sort peripheral blood mononuclear cells from FIV-infected cats into highly (> 95%) purified CD4+ cell, CD8+ cell, immunoglobulin-positive (Ig+) cell, and monocyte subpopulations. The mononuclear cell subpopulations were analyzed for FIV provirus by polymerase chain reaction and Southern blot analysis and for virus expression by RT activity. All 16 cats infected with FIV-NCSU1 demonstrated FIV provirus in CD4+ cell-, CD8+ cell-, and Ig+ cell-enriched lymphocyte populations. Southern blot detection of amplified gag gene sequences and limiting-cell-dilution polymerase chain reaction analysis indicated that Ig+ cells carried a higher FIV provirus burden in chronically (> or = 1-year) infected cats than either CD4+ or CD8+ cells. In contrast, CD4+ cells carried the greatest provirus burden in acutely (2- to 4-week) infected cats. FIV provirus was detected in monocytes from only 1 of 10 cats with asymptomatic infection. Addition of culture supernatants from enriched CD4+, CD8+, and Ig+ cells from FIV-infected cats to an FIV-susceptible CD4+ lymphocyte culture resulted in syncytium formation, cell death, and RT activity. Infection of Ig+ cells is not unique to FIV-NCSU1, as lymphocyte subpopulations from other cats with natural infections and cats infected with the Petaluma or Mount Airy isolate of FIV demonstrated a similar distribution of FIV provirus and RT activity. These data suggest that FIV possesses a broad tropism for peripheral blood mononuclear cells and that an Ig+ cell may serve as a major reservoir for the virus in chronically infected cats.

Animals↗

Pulmonary embolism: diagnosis with electron-beam CT.

Contrast material-enhanced electron-beam computed tomography (CT) (100-msec scan time) was performed to image the pulmonary vasculature in 86 patients, each suspected of having a pulmonary embolism (PE). Thromboembolic material was demonstrated in 39 patients; no emboli were demonstrated in 47. In 25 patients, angiographic or pathologic proof was available. There were 19 proved positive CT scans, four proved negative scans, one false-negative scan, and one false-positive scan. In the 21 patients with CT and angiographic correlation, evidence of thromboembolic disease was seen in 88 vascular zones. Fifty-three zones were positive for PE at both CT and angiography. Eighteen zones were positive for PE at angiography alone, and 17 zones were positive at CT alone. Electron-beam CT is a potentially effective noninvasive means of diagnosing PE.

Adult↗

Morphologic characterization of castor oil-induced colitis in ponies.

Ten ponies (160-250 kg, ages 17 months to 20 years) developed severe diarrhea within 24 hours of castor oil administration (2.5 ml/kg orally). The diarrhea was most severe between 24 and 48 hours post-dosing and subsided by 72 hours. Ponies were euthanatized at 24, 48, and 72 hours post-dosing and intestine was evaluated histologically and ultrastructurally. Twenty-four hours after dosing, the mucosa of the cecum and ventral colon had extensive superficial epithelial erosion and neutrophil infiltration. In the ileum, the epithelium of villous tips was separated from the lamina propria. Scanning electron microscopic examination of the cecal mucosa revealed that basement membranes were exposed in most areas except within necks of crypts. Ultrastructurally, changes in superficial enterocytes of the cecum and ventral colon were characterized by loss of microvilli, distortion of the cytoplasmic terminal web, expansion of the cytoplasmic matrix with formation of precipitates, and widening of intercellular spaces between junctional complexes. Enterocytes located within necks of crypts were flattened along the basement membrane and extended to margins of erosions. Venules within the superficial lamina propria were occluded by fibrin thrombi. Erosions in the cecum and ventral colon of ponies examined 48 hours after treatment were less extensive than those of ponies examined at 24 hours. At 48 hours post-dosing, basement membranes adjacent to crypts were covered by cuboidal enterocytes characterized ultrastructurally by sparse, irregularly shaped microvilli located on broad cytoplasmic protrusions and by numerous free ribosomes. These features indicated that immature enterocytes had migrated from crypts to resurface the eroded mucosa.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Serum ELAM-1 is increased in vasculitis, scleroderma, and systemic lupus erythematosus.

OBJECTIVE: To investigate the state of endothelial cell activation in vasculitis, scleroderma, and systemic lupus erythematosus (SLE). METHODS: We used a sandwich ELISA to quantitate a soluble form of endothelial leukocyte adhesion molecule-1 (sELAM) in serum. RESULTS: sELAM was detected in serum from healthy individuals (mean 0.92 ng/ml). Levels were significantly higher in patients with giant cell arteritis (mean 2.04 ng/ml), polyarteritis nodosa (mean 2.08 ng/ml), scleroderma (mean 2.27 ng/ml), and SLE (mean 3.93 ng/ml). Elevated values were present in patients with both active and inactive disease. sELAM levels of > 3 ng/ml identified most patients with recent onset or active disease. CONCLUSION: Our findings may reflect a low degree of endothelial cell activation in healthy persons that is increased in inflammatory diseases involving blood vessels. Elevated serum sELAM levels may reflect ongoing inflammatory processes in these diseases.

Adolescent↗

Photoelastic stress analysis of natural teeth and three osseointegrated implant designs.

This study used photoelastic stress analysis to compare the location and magnitude of periradicular stresses created by load application directed through two anatomically different natural teeth and three different implant designs. Also studied was the contribution made by the periodontal ligament in the transfer of these stresses. Within the limits of this study, results suggest that the abutment structures with the greatest amount of periodontal support tend to distribute stresses in the most favorable manner. Also, the conical shape of the tooth root, with its large cervical area, has a distinctly positive effect on stress distribution.

Birefringence↗

[The clinical experience in Panama with metronidazole in treating Chagas' disease in the acute and chronic phases].

The authors evaluated the serological response of metronidazole, or 8823 RP-(Hydroxy-2' ethy1)-1-methyl-2 nitro-5 imidazole) in 1307 patients, with Chagas' disease in the acute or chronic stage with age ranging between 6 months to 73 years. 273 patients were in the acute phase: 153 children and 120 adults. Patients in the chronic stage were classified with serological, clinical and cardiological criteria: 1.-POSITIVE SEROLOGY without cardiological signs or symptomatology (EKG, VECTO, ECHO M mode and 2-D, Chest Rx, physiological and pharmacological tests, radiocardiological-HMIBI, MUGA); 2.- POSITIVE SEROLOGY with cardiological signs and symptoms (all cardiological tests were abnormals); 3.- POSITIVE SEROLOGY with cardiomyopathy and cardiac failure. Metronidazole, P.O. or I.V. was utilizing dosages ranging from 15 to 25 mg/Kilo body weight per five days in acute phase and 250 to 500 mg. P.O. in the chronic stage of the disease for 180 days. The drug was well tolerated by all patients in both phases of the disease. Adequate anabolic effect was identified in both stages of the illness. Serological reactions were negative in all patients (100%) without cardiomyopathy. In patients with cardiomyopathy and cardiac failure, the complement fixation test title were similar before and after treatment.

Acute Disease↗

Adherence events in the pathogenesis of infective endocarditis.

Infective endocarditis begins with adherence of microorganisms to cardiac tissues. These tissues have often been previously damaged, creating a thrombotic lesion consisting of platelets and fibrin. Circulating microorganisms localize to this lesion. The tissue specificity of endocarditis likely results from interactions between cell-surface determinants on the endocardium, platelet, and microorganism. Interference with these binding events may offer a means of modifying the course of the infection.

Animals↗

Staphylococcus aureus binding to cardiac endothelial cells is partly mediated by a 130 kilodalton glycoprotein.

Binding of Staphylococcus aureus to vascular endothelial cells may be an initiating event in the pathogenesis of endovascular infection, particularly infective endocarditis. In this study a competition assay between labeled and unlabeled bacteria was used to identify potential S. aureus-binding determinants on the surface of cultured porcine aortic valve endothelial cells. Concanavalin A inhibited 30% to 40% of the specific binding of S. aureus to membrane-associated components. Concanavalin A affinity chromatography of radiolabeled cell-surface proteins, followed by isolation of S. aureus-binding proteins, identified a 130,000-molecular-weight cell surface protein that may function as an endothelial cell S. aureus receptor. These data suggest that specific binding of S. aureus to cardiac valve endothelial cells is mediated in part by a 130 kd mannose-containing glycoprotein.

Animals↗

Fibronectin biosynthesis and cell-surface expression by cardiac and non-cardiac endothelial cells.

We examined the biosynthesis and surface expression of fibronectin, an adhesive glycoprotein, in several types of cultured porcine endothelial cells: pulmonary artery, thoracic aorta, coronary artery, aortic valve, and mitral valve. We used immunocytochemical staining to compare the levels of fibronectin present in these same tissues in vivo. Using endogenous radiolabeling, we found that all cell types except aortic valve endothelial cells synthesized and released into the culture media substantial quantities of fibronectin. Using radioiodination of intact cells, we found that, whereas both thoracic aorta and pulmonary artery cells had measurable fibronectin on the surface, aortic valve, mitral valve, and coronary artery cells had little cell-surface fibronectin present. Immunocytochemical staining showed that all endothelial regions except aortic valve had substantial quantities of immunoreactive fibronectin in vivo. These data suggest that the aortic valve endothelium may be distinct from other endothelia. Such differences could be important for the pathogenesis of valvular disease.

Animals↗

Differential scanning calorimetry of thermal unfolding of the methionine repressor protein (MetJ) from Escherichia coli.

The thermal stability of the methionine repressor protein from Escherichia coli (MetJ) has been examined over a wide range of pH (pH 3.5-10) and ionic strength conditions using differential scanning calorimetry. Under reducing conditions, the transitions are fully reversible, and thermograms are characteristic of the cooperative unfolding of a globular protein with a molecular weight corresponding to the MetJ dimer, indicating that no dissociation of this dimeric protein occurs before unfolding of the polypeptide chains under most conditions. In the absence of reducing agent, repeated scans in the calorimeter show only partial reversibility, though the thermodynamic parameters derived from the first scans are comparable to those obtained under fully reversible conditions. The protein is maximally stable (Tm 58.5 degrees C) at about pH 6, close to the estimated isoelectric point, and stability is enhanced by increasing ionic strength in the range I = 0.01-0.4 M. The average calorimetric transition enthalpy (delta Hm) for the dimer is 505 +/- 28 kJ mol-1 under physiological conditions (pH 7, I = 0.125, Tm = 53.2 degrees C) and shows a small temperature dependence which is consistent with an apparent denaturational heat capacity change (delta Cp) of about +8.9 kJ K-1 mol-1. The effects of both pH and ionic strength on the transition temperature and free energy of MetJ unfolding are inconsistent with any single amino acid contribution and are more likely the result of more general electrostatic interactions, possibly including significant contributions from electrostatic repulsion between the like-charged monomers which can be modeled by a Debye-Hückel screened potential.

Bacterial Proteins↗

Synthesis of renin by tubulocystic epithelium in autosomal-dominant polycystic kidney disease.

Evidence suggests an important role for the renin-angiotensin system in the pathogenesis of autosomal-dominant polycystic kidney disease (ADPKD). Therefore, we studied the presence of immunoreactive renin in renal biopsies and measured the concentrations of renin in cyst fluids. Normal kidneys and kidneys with renal artery stenosis were used for comparison. In ADPKD, immunoreactive renin was present in juxtaglomerular apparatus, associated arterioles, and in some cells within the connective tissue surrounding the cysts. Vascular immunoreactive renin was less prominent than in renal artery stenosis. Increased amounts of tubular immunoreactive renin were noted in polycystic kidneys, as compared to normal kidneys and kidneys with renal artery stenosis. Cyst fluids contained renin detected by Western analysis and enzymatic activity; concentrations were greater in gradient cysts than in nongradient cysts. Seventy-four percent of the renin in gradient cysts was active as compared to 23% in nongradient cysts and 15% in plasma. To determine whether cyst epithelial cells are capable of synthesizing renin, these cells were isolated in tissue culture. Enzymatic assay of extracts from these cells revealed the presence of renin-like enzymatic activity (1.3 +/- 0.8 ng AI/mg protein/hr). The synthesis of renin by tubulocystic epithelium was confirmed by [35S]-methionine radiolabeling of cyst-derived cells, followed by immunoprecipitation and SDS-PAGE and by detection of renin mRNA by the polymerase chain reaction. These results indicate that the tubulocystic epithelium has the potential to synthesize renin. Elevated levels of active renin in renal cysts may be linked to the pathogenesis of hypertension in ADPKD. The occurrence of renin in the lining epithelium of cyst walls raises the possibility that abnormal expression of the renin-angiotensin system may, by a paracrine or autocrine mechanism, regulate epithelial hyperplasia in growing renal cysts.

Base Sequence↗