Down-regulation of regulatory subunit type 1A of protein kinase A leads to endocrine and other tumors

Mutations of the human type Ialpha regulatory subunit (RIalpha) of cyclic AMP-dependent protein kinase (PKA; PRKAR1A) lead to altered kinase activity, primary pigmented nodular adrenocortical disease, and tumors of the thyroid and other tissues. To bypass the early embryonic lethality of Prkar1a(-/-) mice, we established transgenic mice carrying an antisense transgene for Prkar1a exon 2 (X2AS) under the control of a tetracycline-responsive promoter. Down-regulation of Prkar1a by up to 70% was achieved in transgenic mouse tissues and embryonic fibroblasts, with concomitant changes in kinase activity and increased cell proliferation, respectively. Mice developed thyroid follicular hyperplasia and adenomas, adrenocortical hyperplasia, and other features reminiscent of primary pigmented nodular adrenocortical disease, histiocytic and epithelial hyperplasias, lymphomas, and other mesenchymal tumors. These were associated with allelic losses of the mouse chromosome 11 Prkar1a locus, an increase in total type II PKA activity, and higher RIIbeta protein levels. This mouse provides a novel, useful tool for the investigation of cyclic AMP, RIalpha, and PKA functions and confirms the critical role of Prkar1a in tumorigenesis in endocrine and other tissues.     

Glycosaminoglycan profiles of myxomatous mitral leaflets and chordae parallel the severity of mechanical alterations

OBJECTIVES: This biochemical study compared the extracellular matrix of normal mitral valves and myxomatous mitral valves with either unileaflet prolapse (ULP) or bileaflet prolapse (BLP). BACKGROUND: Myxomatous mitral valves are weaker and more extensible than normal valves, and myxomatous chordae are more mechanically compromised than leaflets. Despite histological evidence that glycosaminoglycans (GAGs) accumulate in myxomatous valves, previous biochemical analyses have not adequately examined the different GAG classes. METHODS: Leaflets and chordae from myxomatous valves (n = 41 ULP, 31 BLP) and normal valves (n = 27) were dried, dissolved, and assayed for deoxyribonucleic acid, collagen, and total GAGs. Specific GAG classes were analyzed with selective enzyme digestions and fluorophore-assisted carbohydrate electrophoresis. RESULTS: Biochemical changes were more pronounced in chordae than in leaflets. Myxomatous leaflets and chordae had 3% to 9% more water content and 30% to 150% higher GAG concentrations than normal. Collagen concentration was slightly elevated in the myxomatous valves. Chordae from ULP had 62% more GAGs than those from BLP, primarily from elevated levels of hyaluronan and chondroitin-6-sulfate. CONCLUSIONS: The GAG classes elevated in the myxomatous chordae are associated with matrix microstructure and elastic fiber deficiencies and may influence the hydration-related "floppy" nature of these tissues. These abnormalities may be related to the reported mechanical weakness of myxomatous chordae. The biochemical differences between ULP and BLP confirm previous mechanical and echocardiographic distinctions.     

Fatigue of paralyzed and control thenar muscles induced by variable or constant frequency stimulation

Muscles paralyzed by chronic (>1 yr) spinal cord injury fatigue readily. Our aim was to evaluate whether the fatigability of paralyzed thenar muscles (n = 10) could be reduced by the repeated delivery of variable versus constant frequency pulse trains. Fatigue was induced in four ways. Intermittent supramaximal median nerve stimulation (300-ms-duration trains) was delivered at 1) constant high frequency (13 pulses at 40 Hz each second for 2 min); 2) variable high frequency (each second for 2 min). The first two intervals of each variable frequency train were 5 and 20 ms. The remaining pulses were evenly distributed in time across 275 ms. The number of pulses varied for each subject such that the force time integral in the unfatigued state matched that evoked by a constant 40-Hz train; 3) constant low frequency (7 pulses at 20 Hz each second for 4 min); and 4) variable low frequency (each second for 4 min). The pulse pattern was the same as that for variable high frequency except that the force-time integral was matched to that produced by the constant low-frequency stimulation. These same experiments were performed on the thenar muscles of five able-bodied control subjects. The variable high-frequency trains used to fatigue paralyzed and control muscles had an average (+/- SE) of 12 +/- 2 and 10 +/- 1 pulses, respectively. Variable low-frequency trains had 7 +/- 1 and 6 +/- 1 pulses, respectively. Significant mean force declines of comparable magnitude (to 20-25% initial fatigue force or to 13-21% initial 50 Hz force) were seen in paralyzed muscles with all four stimulation protocols. The force reductions in paralyzed muscles were always accompanied by significant increases in half-relaxation time and decreases in force-time integral, irrespective of the stimulation protocol. Significant force decreases also occurred in control muscles during each fatigue test. Again, these force declines were similar whether constant or variable pulse patterns were used at high or low frequencies (to 40-60% initial fatigue force or to 29-36% initial 50 Hz force). The force reductions in control muscles were significantly less than those seen in paralyzed muscles, except when constant high-frequency stimulation was used. The variations in stimulation frequency, pulse pattern, and pulse number used in this study therefore had little influence on thenar muscle fatigue in control subjects or in spinal cord-injured subjects with chronic paralysis.     

Maternal morbidity and perinatal outcomes among pregnant women with respiratory hospitalizations during influenza season

OBJECTIVES: A population-based assessment of maternal and perinatal morbidity related to respiratory illness during influenza season among pregnant women has not been published. The objectives of this investigation were to describe and quantify the impact of respiratory hospitalization during pregnancy on serious maternal and perinatal morbidity. STUDY DESIGN: A matched cohort study using an administrative database of pregnant women enrolled in the Tennessee Medicaid population to determine pregnancy outcomes associated with respiratory hospitalizations during influenza season. Pregnant women aged 15 to 44 years with a respiratory hospitalization during influenza seasons 1985-1993 were matched by gestational age and presence of comorbidity with pregnant control subjects without a respiratory hospitalization. RESULTS: During the eight influenza seasons studied, 293 women with singleton pregnancies had respiratory disease hospitalizations (5.1:1000). Women with asthma had high rates of such hospitalization (59.7:1000). Compared with matched controls, women with respiratory hospitalizations had similar modes of delivery, delivery length of stay, and episodes of preterm labor. The prevalence of prematurity and low birth weight among infants born to such women was likewise similar between the two groups. CONCLUSION: In this population of pregnant women, those with asthma accounted for half of all respiratory-related hospitalizations during influenza seasons, with 6% of pregnant women with asthma requiring respiratory hospitalization during influenza season, (odds ratio 10.63, 95% CI, 8.18-13.83, compared with women without a medical comorbidity). We detected no significant increase in adverse perinatal outcomes associated with respiratory hospitalizations during influenza season.     

Increased expression of interleukin-9, interleukin-9 receptor,and the calcium-activated chloride channel hCLCA1 in the upper airways of patients with cystic fibrosis

OBJECTIVES/HYPOTHESIS: Mucus overproduction is commonly found in airway disease in patients with cystic fibrosis. Interleukin-9 (IL-9) has been shown to mediate airway hyper-responsiveness and mucus overproduction. Recently, the calcium-activated chloride channel hCLCA1 has been described to be upregulated by IL-9 and has been thought to regulate the expression of soluble gel-forming mucins. We sought to examine the expression of IL-9, interleukin-9 receptor (IL-9R), and hCLCA1 in the upper airway of patients with cystic fibrosis in comparison to healthy control subjects and to demonstrate the relationship of IL-9, IL-9R, and hCLCA1 expression with mucus production. STUDY DESIGN: Prospective design. METHODS: Biopsy samples from nasal polyps of four patients with cystic fibrosis, nasal mucosa of six patients with cystic fibrosis, sinus mucosa of eight patients with cystic fibrosis, and nasal mucosa of six healthy control subjects were stained with periodic acid-Schiff (PAS) to identify mucus glycoconjugates. IL-9, IL-9R, and hCLCA1 expression was determined by immunocytochemical study. RESULTS: We demonstrated significant increases in IL-9, IL-9R, and hCLCA1 immunoreactivity in the mucosa of patients with cystic fibrosis compared with that found in control subjects (P <.05). There were no significant differences between the different locations (nasal polyps, nasal mucosa, and sinus mucosa) in the patient group (P >.05). We also observed a significant increase in the number of mucus-producing cells in biopsy specimens from patients with cystic fibrosis in comparison to control subjects. A positive correlation was found between hCLCA1-positive cells and IL-9-positive cells (correlation coefficient [r] = 0.79, P <.05) or IL-9R-positive cells (r = 0.92, P <.05). Moreover, a positive correlation was also present between PAS-positive (mucus-producing) cells and hCLCA1-positive cells (r = 0.64, P <.05) or IL-9R-positive cells (r = 0.64, P <.05). CONCLUSIONS: Increased expression of IL-9 and IL-9R, as well as upregulation of hCLCA1, in mucus-overproducing epithelium of patients with cystic fibrosis supports the hypothesis that IL-9 contributes to mucus overproduction in cystic fibrosis. Expression of hCLCA1 may also be responsible, in part, for the overproduction of mucus. These preliminary findings suggest that hCLCA1 might be an interesting new therapeutic target to control mucus overproduction in airway disease in patients with cystic fibrosis.     

Effect of orange and apple juices on iron absorption in children

OBJECTIVE: To measure iron absorption in children from meals containing apple juice or orange juice so as to determine if iron absorption will be greater with orange juice because of its higher ascorbic acid content than apple juice, a noncitrus fruit juice that US children reportedly prefer. DESIGN: On 2 successive days, children consumed identical meals that included apple juice on one day and orange juice on the other, in random order. The meals were labeled with iron-57 on one day and iron-58 on the other. Iron absorption was measured from red blood cell incorporation of the iron stable isotopes 14 days later. SETTING: Nutrition research institute in a major metropolitan medical center. PATIENTS: A total of 25 healthy children, 3 to 6 years of age, were recruited, of whom 21 (11 male and 10 female) completed the study.Intervention Identical meals served with orange juice and apple juice were given on consecutive days, in a balanced randomized design. MAIN OUTCOME MEASURES: Iron absorption measured by established stable isotope methods. RESULTS: Median iron absorption from the meal ingested with apple juice was 7.17% (mean +/- SD, 9.48% +/- 9.68%). Median iron absorption from the meal ingested with orange juice was 7.78% (9.80% +/- 6.66%; P =.44). Iron absorption from the meal that included apple juice was significantly correlated with serum ferritin concentration (P =.02); iron absorption from the meal that included orange juice tended to correlate with serum transferrin receptor concentration (P =.051). CONCLUSIONS: As children absorb iron well from a meal that includes either orange or apple juice, a preference for apple juice does not pose a concern with regard to the prospect of iron-deficiency anemia, which remains a significant health problem in the United States.     

Toxic neuropathy in patients with pre-existing neuropathy

The authors report significant worsening of a pre-existing neuropathy in six patients who received "non-toxic" dosages of known neurotoxic agents. Before treatment, baseline total neuropathy score (TNS) averaged 9.5 (range 0 to 19). After chemotherapy (Taxol [125 to 175 mg/m(2) x 4]; vincristine [2 to 5 mg]; cisplatin [40 mg/m(2) x 8]; and thalidomide [60 g]), the TNS averaged 22 (range 13 to 29). The authors conclude that functionally disabling toxic neuropathy can occur in patients with pre-existing neuropathy at standard doses.     

Myelin basic protein-reactive autoantibodies in the serum and cerebrospinal fluid of multiple sclerosis patients are characterized by low-affinity interactions

The presence of autoantibodies to the immunodominant antigen, myelin basic protein (MBP), in the serum and cerebrospinal fluid (CSF) of patients with multiple sclerosis (MS) has been poorly characterized. Many studies report detectable levels of autoantibodies to myelin basic protein though other studies, using similar techniques, report their absence. We compared a solution-phase assay that has detected clinically relevant autoantibodies in diabetes and other autoimmune diseases to solid phase assays similar to those used in previous reports. The solution-phase assay consistently measured autoantibodies to MBP in serum from human subjects with Semple rabies vaccine (SRV)-induced demyelinating disease and from MBP-immunized animals. A solid phase assay detected MBP autoantibodies in the serum of a fraction of patients with MS. Autoantibodies capable of binding to MBP in the solution-phase were not detected in the CSF or serum of patients with MS. Additional solution-phase measurements revealed that anti-MBP antibodies from individuals with SRV-induced demyelinating disease demonstrated a binding affinity profile consistent with that of polyclonal antibodies with a range of affinities from low to high. In contrast, antibodies to MBP in the serum of MS patients detected by ELISA did not bind soluble MBP in the same assay. These results indicate that the humoral response in patients with MS does not include moderate- or high-affinity autoantibodies to MBP.