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Biomedical subjects

R J Johnson

Publications and source records attributed to R J Johnson.

At least 199 records · Page 11Linked to original sources
PubMed1994

Decreasing calreticulin expression lowers the Ca2+ response to bradykinin and increases sensitivity to ionomycin in NG-108-15 cells.

It has been suggested that the multifunctional protein, calreticulin, is a major calcium sequestering protein in the inositol 1,4,5-trisphosphate receptor-containing endoplasmic reticulum subcompartment. In neuroblastoma X glioma NG-108-15 cells, bradykinin can effectively stimulate the release of inositol 1,4,5-trisphosphate and cause a cytosolic calcium transient. To explore the function of calreticulin as an intracellular calcium sequestering protein, we investigated calcium dynamics in NG-108-15 cells after treatment with an antisense oligonucleotide against calreticulin, CrtAS1. Cells treated with either CrtAS1 or the corresponding sense oligonucleotide CrtPS1 were examined for their calreticulin content by Western blotting, the amplitude of their calcium transient in response to bradykinin, and their sensitivity toward the calcium ionophore, ionomycin. Treatment with CrtAS1 decreased the amount of calreticulin in comparison to CrtPS1-treated and untreated control cells. At the same time, CrtAS1-treated cells had a significantly reduced calcium response to bradykinin and were more sensitive to ionomycin-induced cell death. These data show that the level of calreticulin expression is directly related to the calcium storage capacity of the inositol 1,4,5-trisphosphate-sensitive calcium pool and indicate a direct relationship between the level of calreticulin and the protection against cytotoxic calcium overload.

Base Sequence↗
PubMed1994

Determination of a zero strain reference for the anteromedial band of the anterior cruciate ligament.

The objective of this study was to verify a method previously used to determine a reference length for calculations of anterior cruciate ligament strain. In nine knee specimens, an arthroscopic force probe and a Hall effect transducer were placed in the anteromedial band of the ligament. Anteroposterior-directed shear loads then were applied to the knee joint with the knee flexed to 30 degrees. From the sigmoidal curve for shear load versus displacement of the anterior cruciate ligament midsubstance, the length of the transducer at the inflection point was determined graphically by two independent examiners. Previous studies suggested that the inflection point corresponds to the slack-taut transition of the anteromedial band. The force probe was used to determine the actual length of the transducer when the anteromedial band became load bearing. No significant differences were found between the reference lengths determined by the inflection point method and the force probe. The force probe demonstrated that the anterior cruciate ligament became load bearing when an anterior shear load of 8.8 N was applied to the tibia with the knee at 30 degrees of flexion. Furthermore, multiple cycles of anteroposterior shear loading did not influence these values. The force probe verified that the inflection method provides a reasonable estimate of the absolute strain reference (within 0.7% strain).

Adult↗
PubMed1994

Staging, volume estimation and assessment of nodal status in carcinoma of the cervix: comparison of magnetic resonance imaging with surgical findings.

Pre-operative magnetic resonance imaging (MRI) was carried out in 50 women scheduled for operative treatment of invasive carcinoma of the cervix. The extent of the primary tumour (stage), its dimensions and the presence of lymph node enlargement were assessed and compared with findings at surgery and/or histopathological examination of the resected uterus. In 45 patients undergoing radical hysterectomy, accuracy of MRI staging of the primary tumour was 84.4%. In the group as a whole, including four patients with inoperable disease, staging accuracy was 84%. Most errors were due to difficulty in identifying early vaginal or parametrial invasion by tumour. There was close correlation between the volume of tumour measured from pre-operative MRI scans and measurements made on the hysterectomy specimen (r = 0.95). MRI had a sensitivity of 75% and a specificity of 88% in predicting metastatic lymphadenopathy, based solely on the criterion of enlargement of any pelvic or para-aortic nodes to 1.5 cm or greater. However, retrospective analysis of the presence or absence of metastases by site in 49 patients undergoing lymphadenectomy or lymph node sampling at laparotomy showed that true sensitivity to be 57.1% and the specificity 96.8%. Differentiation between malignant and reactive lymphadenopathy was not reliably achieved on MRI, and in several patients, metastases were present in normal-sized lymph nodes.

Adolescent↗
PubMed1994

Mechanisms of progressive renal disease in glomerulonephritis.

Progressive renal disease in glomerulonephritis (GN) involves both glomerular and interstitial processes. In the glomerulus, sclerosis occurs with progressive accumulation of extracellular matrix components that reduce filtration surface area. In the interstitium, early inflammatory changes accompany GN with later development of fibrosis and tubular atrophy. Our studies have focused on the role of early cellular events in the development of glomerular and interstitial fibrosis. In the antithymocyte serum (ATS) model of mesangial proliferative GN, mesangial cell proliferation is initiated by processes involving complement and platelets and may involve basic fibroblast growth factor (bFGF). Mesangial cell proliferation is maintained by an autocrine mechanism involving upregulation of mesangial cell PDGF and PDGF receptors. Mesangial cells also change phenotype with expression of alpha-smooth muscle actin and production of type I collagen. These early changes precede upregulation of genes for the production of extracellular matrix components and the development of mesangial matrix expansion and sclerosis. Matrix expansion is reduced by factors that block cell proliferation, including platelet and complement depletion, heparin, and antibody to PDGF. A similar sequence of early platelet infiltration, increased expression of PDGF, and mesangial cell proliferation occurs early in the development of glomerulosclerosis in the remnant kidney model, and mesangial cell proliferation is a prominent early feature of experimental diabetic nephropathy. We believe these early glomerular cellular changes are linked to the later development of sclerosis. In the interstitium, acute GN is accompanied by upregulation of mRNA and protein for osteopontin, a macrophage chemotactic/adhesive factor expressed by cortical tubules following several types of glomerular injury.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

C5b-9 increases albumin permeability of isolated glomeruli in vitro.

Deposition of antibody and activation of the complement cascade are important in both naturally occurring glomerulonephritis and in experimental models including passive Heymann nephritis. We studied the effect of antibody and complement on albumin permeability of isolated glomeruli to determine the role of the terminal complement components (C5-C9) in mediating the proteinuria in nephritis. Isolated glomeruli were treated with anti-Fx1a (Heymann antibody) and then incubated them with pooled human serum, serum in which complement had been inactivated by heat, or serum deficient in C6 or C7. The albumin reflection coefficient (sigma albumin) was calculated from the volumetric response of glomeruli to transcapillary oncotic gradients produced by albumin or high molecular weight neutral dextran (252 kD). Convectional permeability to albumin (Palbumin) was calculated as 1-sigma albumin. Albumin permeability of control glomeruli was not different from 0. Albumin permeability was not altered by antibody alone but was increased to 0.65 +/- 0.04 when antibody treated glomeruli were incubated for 10 minutes with pooled serum as a source of complement. Heat treatment of serum to inactivate complement prevented the increase in permeability. Incubation for 10 minutes with serum without antibody pretreatment caused a lesser increase in permeability of isolated glomeruli (0.18 +/- 0.06). Serum deficient in either C6 or C7 did not cause an increase in albumin permeability of antibody pre-treated glomeruli, but incubation with a combination of these sera (now containing the complete cascade) increased permeability to the same extent as did pooled normal serum (0.58 +/- 0.04).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Hepatitis C virus-associated glomerulonephritis. Effect of alpha-interferon therapy.

Hepatitis C virus (HCV) infection may present as a primary glomerular disease. We report 34 adult patients who presented with proteinuria and had circulating anti-HCV antibodies. Primary risk factors included a history of intravenous drug abuse (56%) or blood transfusion (18%). Patients presented with nephrotic syndrome (71%) or with non-nephrotic proteinuria (29%) and had membranoproliferative or acute proliferative glomerulonephritis on renal biopsy. Signs of clinical liver disease were infrequent (18%), though elevated liver function tests were common (66%) and liver biopsy in 16 of 18 patients showed chronic active hepatitis. Cryoglobulinemia was frequent (59%), but only 44% had extrarenal manifestations. In 100% of cases tested, HCV RNA could be found in the serum or cryoprecipitates. Fourteen patients received interferon alpha for 6 to 12 months with a significant reduction in proteinuria but no improvement in renal function. A good clinical response correlated with disappearance of HCV RNA from the serum during treatment; however, relapse of viremia and renal disease was common after completing therapy. Evidence for HCV infection should be sought in all patients with primary glomerular disease. The optimal treatment strategy, however, remains to be defined.

Adult↗
PubMed1994

Osteopontin expression in angiotensin II-induced tubulointerstitial nephritis.

Osteopontin is an arginine-glycine-aspartate (RGD) containing secreted phosphoprotein recently shown to stimulate a local macrophage influx when injected subcutaneously in mice. We examined the effect of angiotensin II infusion on renal injury and osteopontin expression in the rat kidney by in situ hybridization and immunohistochemistry. Preceding pathologic changes in tubular and interstitial cells, a dramatic increase in renal osteopontin protein and mRNA levels was observed primarily in epithelial cells of the distal tubules, collecting ducts and Bowman's capsule. Although both cortex and medulla showed increased osteopontin levels, the effect was most pronounced in the renal cortex which normally showed very little constitutive osteopontin expression. Interestingly, regions of the kidney expressing high osteopontin levels correlated with sites of monocyte/macrophage accumulation. These observations, coupled with recent findings that osteopontin may be a pro-inflammatory protein, suggests that osteopontin over-expression may facilitate monocyte/macrophage accumulation at the sites of renal tubulointerstitial injury.

Angiotensin II↗
PubMed1994

Renal proliferative and phenotypic changes in rats with two-kidney, one-clip Goldblatt hypertension.

Angiotensin II (AII) is a vasoconstrictive peptide with hypertrophic and mitogenic effects on many cell types. Previous studies have shown that in vivo administration of AII in rats results in proliferation of, and phenotypic changes in, many renal cell populations, but in doses also causing hypertension. Thus, it was not possible to differentiate nonhemodynamic from hypertensive effects of AII. Therefore, we studied rats with renin-dependent, AII-mediated hypertension (the two-kidney, one-clip Goldblatt model; mean systolic blood pressure 238 +/- 48 v 140 +/- 6 mm Hg in sham-operated controls). The unclipped kidneys, which were exposed to high blood pressure, developed significant glomerular and tubulointerstitial injury, tubulointerstitial cell proliferation, dense focal interstitial monocyte-macrophage influx, increased deposition of types I and IV collagen, as well as increased cellular expression of desmin and actin, in tubulointerstitial areas when examined at 11 weeks. In contrast, clipped kidneys, protected from hypertension but with high local renin expression, had minimal abnormalities. These studies suggest that in this model increased renin, and presumably AII, does not mediate significant proliferative or phenotypic changes in the kidney in the absence of hypertension at 11 weeks.

Angiotensin II↗
PubMed1994

Hospital resource consumption among older adults: a prospective analysis of episodes, length of stay, and charges over a seven-year period.

After linking their administrative records and interview data, the consumption of Medicare-reimbursed hospital resources during 1984 through 1990 by the 7,527 LSOA respondents was prospectively assessed using a two-part design. First, logistic regression was used to model whether a hospital episode occurred. Second, among those having had hospital episodes, OLS regression was used to model the number of episodes, as well as the natural logarithms of the total length of stay and the total charges. The risk of hospitalization was mostly associated with being male, prior hospital and physician utilization, and lower body limitations. Among those hospitalized: (a) greater numbers of episodes were mostly associated with prior hospital and physician utilization, and poorer perceived health; (b) longer lengths of stay were mostly associated with prior hospital and physician utilization, and poorer perceived health; and, (c) higher charges were mostly associated with population density, poorer perceived health, and prior physician and hospital utilization. Decedents consistently consumed substantially more hospital resources than survivors.

Activities of Daily Living↗
PubMed1994

Gender, race, and health: the structure of health status among older adults.

A previously developed model of disease, disability, functional limitation, and perceived health was examined for race and/or gender biases. This model focuses on (a) the direct effects of three factors on perceived health status, (b) how disability, functional limitations, and self-rated health interrelate, and (c) how race and gender condition these interrelationships. The results confirm the construct validity of separate dimensions of disability and functional limitation, and indicate that their differential effects are further modified by gender. Eight significant differences in structural effects are identified, including one gender effect among both blacks and whites, and seven additional gender effects among whites. In the structural model, then, most differences are gender differences among whites. The significant racial differences within gender were found only in the measurement model. Race differences for upper body disability and perceived health are consistent across gender. Sex differences, however, in measures of basic ADLs and household ADLs are not consistent across race. The findings confirm earlier conclusions that differences in the measurement of health exist between males and females, and between blacks and whites, but that the differences in the causes of perceived health exist only between males and females.

Aged↗
PubMed1994

Effect of PAF on rat lung vascular permeability: role of platelets and polymorphonuclear leucocytes.

1. The objectives of the present experiments were to assess the contribution of polymorphonuclear leucocytes (PMNLs), platelets and their products such as thromboxane A2 (TxA2), histamine and 5-hydroxytryptamine to platelet activating factor (PAF)-mediated protein extravasation in rat lungs. 2. Intravenous injection of PAF (1.0 and 5.0 micrograms kg-1) increased dose-dependently (up to 7.5 fold) the vascular permeability of the trachea, upper and lower bronchi to Evans blue dye (EB), a marker of albumin extravasation. The permeability of the pulmonary parenchyma was not affected significantly by PAF. 3. Thrombocytopenia induced by administration of the IgG fraction of goat anti-rat platelet serum (APS; 15 mg 100 g-1, i.p., 16-18 h) reduced by 55, 58 and 40% the effects of the lower dose of PAF (1.0 microgram kg-1) and by 31, 23 and 15% the effects of the higher dose of PAF (5.0 micrograms kg-1) on the permeability of the trachea, upper and lower bronchi respectively to albumin. 4. PMNL depletion induced by administration of rabbit anti-rat polymorphonuclear serum (ANS; 2 mg kg-1, i.v., 24 h) did not reduce significantly the effects of the lower dose of PAF (1.0 microgram kg-1) on the airways, however the effects of the higher dose of PAF (5.0 micrograms kg-1) on the permeability of the trachea, upper and lower bronchi to albumin were reduced by 43, 25 and 23% respectively. 5. The injection of both the anti-platelet and the anti-PMNL sera reduced by 61, 62 and 96% the effects of the lower dose of PAF (1.0 microg kg-1) and by 44, 39 and 47% the effects of the higher dose of PAF (5.0 microg kg-1) on the permeability of the trachea, upper and lower bronchi respectively.6. The combined injection of the TxA2-mimetic (U-44069; 5.0 microg kg-1) and PAF (1.0 and 5.0 microg kg-1)in thrombocytopenic rats overcame the vascular permeability decrease induced by APS treatment.7. Pretreatment of the animals with a combination of antagonists to histamine (mepyramine;3.0 mg kg-1) and 5-hydroxytryptamine (methysergide; 2.5 mg kg-1) did not cause a significant inhibition of the effect of PAF (1.0 and 5.0 microg kg-1) on EB extravasation in the airways.8. These data show that the effect of intravenous PAF on rat vascular permeability is partly modulated by polymorphonuclear leucocyte and platelet activation. Our results suggest that following its release,TxA2 could increase postcapillary hydrostatic pressure by inducing a venoconstriction and potentiate the extravasation elicited by PAF. These results do not suggest a major role for histamine and/or 5-hydroxytryptamine on PAF-induced albumin extravasation.

Animals↗
PubMed1994

Vibrio cholerae O139 synonym bengal is closely related to Vibrio cholerae El Tor but has important differences.

Although Vibrio cholerae O139 synonym Bengal strains, from the current epidemics in India and Bangladesh, are closely related to seventh-pandemic strains, as shown by multilocus enzyme electrophoresis, Bengal strains are encapsulated and portions of the O1 antigen biosynthetic complex genes found in O1 strains are altered or lacking. Encapsulated Bengal strains showed resistance to killing by normal human serum. The presence of the capsule suggests the potential for bloodstream invasion in susceptible hosts and has profound implications for vaccine development.

Animals↗