Differential effect of phorbol esters and interleukin-3 on protein kinase C isoform content and kinase activity in the FDC-P1 cell line

FD/PMA is a subclone of the interleukin-3 (IL-3)-dependent, FDC-P1 cell line, which proliferates in response to either 12-O- tetradecanoylphorbol-13 acetate (PMA) or IL-3. While several endogenous substrates were phosphorylated in response to protein kinase C (PKC) activation in FDC-P1, phospholipid-dependent phosphorylation in the FD/PMA grown in PMA was not observed. Basal, phosphatidylserine- independent, and diolein-independent phosphorylation of cytosolic substrates with molecular weights of 17, 52, 57, and 105 Kd were enhanced in FD/PMA cells grown in PMA as compared with FDC-P1 cells cultured in IL-3. Phosphorylation of a 105-Kd substrate was enhanced in the particulate fraction of FD/PMA cells maintained in PMA. The 17-Kd substrate in FD/PMA cells comigrated with a substrate phosphorylated in a PKC-dependent manner in FDC-P1 cells. Phosphorylation of the 52- and 57-Kd substrates, but not of the 17-Kd substrate, was inhibited by H-7 and staurosporine. A portion of the PMA-induced cytosolic kinase activity coeluted with PKC on diethyl aminoethyl chromatography. While FD/PMA cells cultured in PMA contained negligible PKC-dependent phosphorylation of endogenous substrates or histone, alpha and epsilon PKC isoforms were detected by Western blot analysis. PKC phosphotransferase activity was observed in FD/PMA cells grown in PMA when peptides corresponding to residues 720 to 737 of PKC-epsilon or residues 4 to 14 of myelin basic protein were used as substrates. These data indicate that maintenance of FD/PMA cells in PMA stimulates proliferation and markedly alters PKC substrate specificity. Generation of at least two phospholipid-independent kinases occurs in PMA-treated cells.     

Acute lower respiratory illness in childhood as a predictor of lung function and chronic respiratory symptoms

This study investigated the relationship of acute lower respiratory illness (LRI) to level and change in level of forced expiratory volumes in a cohort of 801 children, followed longitudinally for a maximum of 13 yr. The co-occurrence of respiratory illness before 2 yr of age and two or more LRI during a single surveillance year was associated with a 20.3% lower mean cross-sectional level of FEF25-75, and with reduced longitudinal change in level of FEF25-75. The effect of LRI on lung function was uniformly stronger for boys than for girls. Of the children with illness before 2 yr of age and two or more LRI, six of 14 were male asthmatics with mean levels of FEF25-75 that were lower than those of other asthmatic children. Pneumonia and/or hospitalization for respiratory illness prior to the onset of study were associated with lower cross-sectional levels of forced expiratory volumes at entry to the study, even when asthmatics/persistent wheezers were eliminated from the analysis (6.1% lower level of FEV1 for a nonasthmatic boy with previous hospitalization versus a nonasthmatic boy without hospitalization). In the longitudinal analysis, pneumonia and/or hospitalization were associated with slower increase in level of forced expiratory volumes, even after adjusting for "ever diagnosis of asthma/current any wheeze" (starting at the same leve, after eight years a boy with hospitalization would develop a 5.0% lower FEV1 than a boy without hospitalization). Acute LRI also was evaluated as a predictor of chronic respiratory symptoms.(ABSTRACT TRUNCATED AT 250 WORDS)     

Immediate results and long-term follow-up after repeat balloon aortic valvuloplasty

Balloon aortic valvuloplasty (BAV) was performed in 219 elderly patients with aortic stenosis between December 1985 and April 1990. Forty-three patients underwent repeat BAV for symptomatic restenosis of the aortic valve 13±8 mo following initial BAV. To evaluate the outcome following initial and repeat BAV, hemodynamic results were analyzed according to the following subgroups: BAV 1-initial BAV for all patients (n = 219); BAV 1/1-initial BAV in those who had only one BAV (n = 176); BAV 1/2-the initial BAV in those who had repeat BAV (n = 43); and BAV 2-repeat BAV (n = 43). The mean age of patients undergoing BAV 2 was 82±6 yr compared to 78±10 yr for all patients undergoing BAV 1 (p = .01). At the time of BAV 1 there was no difference in baseline or post-valvuloplasty aortic valve area (AVA) or peak aortic valve gradient (AVG) for patients having BAV 1/1 compared to those having BAV 1/2. However, for patients having repeat BAV, although the magnitude of the hemodynamic improvement of BAV 1/2 (AVA increased from 0.6 to 0.9 cm², AVG decreased from 65 to 34 mm Hg, p<.001) was similar to the magnitude of the hemodynamic improvement of BAV 2 (AVA increased from 0.5 to 0.8 cm², AVG decreased from 65 to 34 mm Hg, p<.001), the baseline AVA (0.5 cm² at BAV 2 vs. 0.6 at BAV 1/2) and the post-valvuloplasty AVA (0.8 cm² at BAV 2 vs. 0.9 at BAV 1/2) were significantly smaller (p<.004). There were no major complications in patients undergoing repeat BAV. Median event-free survival was 12 mo after initial BAV and 8 mo after repeat BAV (p = .09). In conclusion, repeat BAV results in similar immediate hemodynamic improvement and event-free survival compared to initial BAV. However, baseline aortic valve area and gradient prior to repeat BAV are significantly worse than those prior to initial BAV, suggesting that balloon-induced injury to the aortic valve may offer only a temporary hiatus from the natural history of aortic stenosis.     

Report of the National Institutes of Health Task Force on Research Standards for Chronic Low Back Pain

Objectives

Despite rapidly increasing intervention, functional disability due to chronic low back pain (cLBP) has increased in recent decades. We often cannot identify mechanisms to explain the major negative impact cLBP has on patients' lives. Such cLBP is often termed nonspecific and may be due to multiple biologic and behavioral etiologies. Researchers use varied inclusion criteria, definitions, baseline assessments, and outcome measures, which impede comparisons and consensus. The purpose of this article is to disseminate the report of the National Institutes of Health (NIH) task force on research standards for cLBP.

Methods

The NIH Pain Consortium charged a research task force (RTF) to draft standards for research on cLBP. The resulting multidisciplinary panel developed a 3-stage process, each with a 2-day meeting.

Results

The panel recommended using 2 questions to define cLBP; classifying cLBP by its impact (defined by pain intensity, pain interference, and physical function); use of a minimal data set to describe research subjects (drawing heavily on the Patient Reported Outcomes Measurement Information System methodology); reporting “responder analyses” in addition to mean outcome scores; and suggestions for future research and dissemination. The Pain Consortium has approved these recommendations, which investigators should incorporate into NIH grant proposals.

Conclusions

The RTF believes that these recommendations will advance the field, help to resolve controversies, and facilitate future research addressing the genomic, neurologic, and other mechanistic substrates of cLBP. Greater consistency in reporting should facilitate comparisons among studies and the development of phenotypes. We expect the RTF recommendations will become a dynamic document and undergo continual improvement.     

Factors Associated With Microalbuminuria in 7,549 Children and Adolescents With Type 1 Diabetes in the T1D Exchange Clinic Registry

OBJECTIVE

To examine factors associated with clinical microalbuminuria (MA) diagnosis in children and adolescents in the T1D Exchange clinic registry.

RESEARCH DESIGN AND METHODS

T1D Exchange participants <20 years of age with type 1 diabetes ≥1 year and urinary albumin-to-creatinine ratio (ACR) measured within the prior 2 years were included in the analysis. MA diagnosis required all of the following: 1) a clinical diagnosis of sustained MA or macroalbuminuria, 2) confirmation of MA diagnosis by either the most recent ACR being ≥30 mg/g or current treatment with an ACE inhibitor (ACEI) or angiotensin receptor blocker (ARB), and 3) no known cause for nephropathy other than diabetes. Logistic regression was used to assess factors associated with MA.

RESULTS

MA was present in 329 of 7,549 (4.4%) participants, with a higher frequency associated with longer diabetes duration, higher mean glycosylated hemoglobin (HbA_1c) level, older age, female sex, higher diastolic blood pressure (BP), and lower BMI (P ≤ 0.01 for each in multivariate analysis). Older age was most strongly associated with MA among participants with HbA_1c ≥9.5% (≥80 mmol/mol). MA was uncommon (<2%) among participants with HbA_1c <7.5% (<58 mmol/mol). Of those with MA, only 36% were receiving ACEI/ARB treatment.

CONCLUSIONS

Our results emphasize the importance of good glycemic and BP control, particularly as diabetes duration increases, in order to reduce the risk of nephropathy. Since age and diabetes duration are important nonmodifiable factors associated with MA, the importance of routine screening is underscored to ensure early diagnosis and timely treatment of MA.Elevated urinary albumin excretion is an early sign of diabetic kidney disease (DKD). The American Diabetes Association (ADA) recommends screening for microalbuminuria (MA) annually in people with type 1 diabetes after 10 years of age and 5 years of diabetes duration, with a diagnosis of MA requiring two of three tests to be abnormal (1). Early diagnosis of MA is important because effective treatments exist to limit the progression of DKD (1). However, although reduced rates of MA have been reported over the past few decades in some (2–4) but not all (5,6) studies, it has been suggested that the development of proteinuria has not been prevented but, rather, has been delayed by ∼10 years and that further improvements in care are needed (7).Limited data exist on the frequency of a clinical diagnosis of MA in the pediatric population with type 1 diabetes in the U.S. Our aim was to use the data from the T1D Exchange clinic registry to assess factors associated with MA in 7,549 children and adolescents with type 1 diabetes.     

Orthotopic cardiac transplantation: evaluation with CT

As cardiac transplantation has become widely available, computed tomography (CT) of the chest has played a useful role in the examination of patients after heart transplantation. To determine anatomic features related to the procedure, the authors evaluated 59 scans in 46 patients who had undergone orthotopic cardiac transplantation. Aortic anastomosis (seen in 98% of scans) and altered spacing between the great vessels (83%) proved to be the most common and most reliable findings. Other features including atrial anastomosis, high main pulmonary artery segment, remnant superior vena cava, and cardiac reorientation were also seen. Accurate interpretation of adenopathy, mediastinal abscess, and pericardial effusion will be enhanced in these patients through a better understanding of the cardiovascular-pericardial complex, which is afforded by CT.