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J Pfeifle

Publications and source records attributed to J Pfeifle.

18 recordsLinked to original sources
PubMed1994

Comparison of intrapulmonary percussive ventilation and chest physiotherapy. A pilot study in patients with cystic fibrosis.

STUDY OBJECTIVE: To compare the intrapulmonary percussive ventilator (IPV) to chest physiotherapy (P&PD) with respect to acute changes in (1) pulmonary function and (2) sputum physical properties in patients with cystic fibrosis (CF). DESIGN: Randomized crossover. SETTING: Community-based CF referral center. PARTICIPANTS: Nine nonhospitalized person (range, 7 to 40 years; median, 12.4 years) with moderate to excellent Shwachman scores. INTERVENTIONS: Three treatment regimens: (1) 2.5 mg albuterol delivered via IPV (internal percussive component activated); (2) 2.5 mg. albuterol delivered via IPV (internal percussive component inactivated), followed by P&PD; and (3) 2.5 mg albuterol delivered via updraft nebulizer, followed by P&PD. MEASUREMENTS AND RESULTS: Outcome measures included pulmonary function testing (PFTs) and quantitative and qualitative sputum analysis. Among the three treatment groups, there were no significant differences in the change in predicted PFTs 1 h or 4 h after treatment, nor in the volume of sputum expectorated in the first 4 or in the subsequent 20 h. Among patients receiving IPV, more serious disease was associated with greater improvement in FEF25-75 1 h after treatment, but these differences disappeared by 4 h. There were no meaningful differences in viscoelastic characteristics of sputum expectorated after each treatments. Participants reported general satisfaction with no adverse effects while using IPV. CONCLUSIONS: This initial pilot study suggests (1) stable patients with CF tolerated one treatment of IPV without adverse sequelae, and (2) IPV was as effective as standard aerosol and P&PD in improving short-term PFT results and enhancing sputum expectoration.

Adolescent↗
PubMed1990

Pathogenesis of paraneoplastic follicular hyperkeratotic spicules in multiple myeloma. Follicular and epidermal accumulation of IgG dysprotein and cryoglobulin.

We describe a 62-year-old man with multiple myeloma who developed horny spicules on his face, particularly on his nose. IgG-lambda monoclonal gammopathy was detected, and the serum dysprotein was shown to be a cryoglobulin, which forms a cryogel at low temperatures. Light and electron microscopic and immunohistochemical examinations showed an intercellular precipitation and massive accumulation of the IgG dysprotein and cryoglobulin between the keratinocytes of the upper epidermis and the infundibular epithelium. The follicles were dilated and filled with parakeratotic cells, the protein deposits between them and a rudimentary hair thus resulting in the clinically visible symptoms of horny spicules. The limitation or the predominance of the symptoms in cold-exposed body regions, the morphological identification of the dysprotein deposits as cryoglobulin or cryogel, and the laboratory findings concerning the temperature and pH dependence of the precipitation of the IgG dysprotein reveal that the paraneoplastic horny spicules are a hitherto unknown clinical manifestation of cryoglobulinemia.

Cryoglobulinemia↗
PubMed1987

Human SS-B/LA autoantigen contains a covalent protein-RNA linkage.

The chemical nature of association of RNA in immunoprecipitates of human SS-B/La ribonucleoprotein, an autoantigen expressed in various autoimmune disorders, was investigated. A fraction of RNA associated with SS-B/La immunoprecipitates was readily dissociated by SDS-polyacrylamide gel electrophoresis, yielding four main subfractions, R1-4, with chain lengths in the range of 90-130 nucleotides (R4), 140-175 nucleotides (R2 and R3) and above 200 nucleotides (R1). Moreover, the immunoreactive protein component, migrating with a molecular mass of 49 kDa, contained a very tightly bound RNA co-migrating with the protein unless the protein was proteolytically degraded. Most of the RNA molecules in this fraction, represented by about 20 components, had a free 3'-terminus but a blocked 5'-terminus and showed chain lengths between 10 and 125 nucleotides. After pretreatment with alkaline phosphatase and a mixture of ribonucleases T1 + T2 + A, adenosine 3',5'-biphosphate (pAp) was liberated by phosphodiesterase (Crotalus durissus) as the blocked 5'-end of the RNA. The chemical nature of the blockage was revealed after alternative treatment of the protein-pAp component with phosphodiesterase or nuclease S7 followed by acid hydrolysis and phosphoamino acid analysis which showed that a threonine residue must be directly involved in the RNA-protein linkage of 49 kDa SS/La antigen, indicating the presence of a covalent threonine-pAp bond.

Alkaline Phosphatase↗
PubMed1987

Multiple phosphorylation of human SS-B/LA autoantigen and its effect on poly(U) and autoantibody binding.

The metabolic turnover rates and the effect of in vitro phosphorylation on poly(U) and autoantibody binding of human SS-B/La ribonucleoprotein, an autoantigen expressed in various autoimmune disorders, were studied. The determination of the metabolic turnover rates of SS-B/La protein, SS-B/La protein phosphorylation and RNA binding yielded values of 12.1 h, 3.6 h and 3.7 h, respectively, indicating a possible functional correlation of RNA-binding and phosphorylation. This assumption was confirmed by studies of in vitro phosphorylation using purified SS-B/La protein and purified casein kinase type II as a model system. A high degree of phosphorylation of the SS-B/La protein (molecular mass 49 kDa) substantially diminished its binding capacity for poly[3H]U. However, binding of human autoantibodies against SS-B/La antigen increases 2-fold with increased SS-B/La phosphorylation. Complete phosphorylation in vitro led to partial molecular transformation, yielding an antigenically cross-reacting component with an apparent molecular mass of 51 kDa which could not be detected during in vivo phosphorylation.

Autoantibodies↗
PubMed1987

Quantitation and potential function of nucleolar phosphoprotein pp 105 in mouse tumor cells, embryonic cells and normal tissues.

1. Nucleolar phosphoprotein pp 105 was determined in various mouse cell and tissue extracts using a highly sensitive ELISA. The results indicate that the highest relative amounts of pp 105 correlate with cells and tissues of high growth rate such as tumor cell lines, solid tumors and embryonic tissues. 2. The specific phosphorylation of pp 105 was compared in a 1 min endogenous phosphorylation assay with native cell and tissue extracts. 3. The mitogenic activity of highly purified pp 105 was demonstrated in cultures of resting mouse embryonic cells and mouse thymocytes.

Animals↗
PubMed1986

Phosphoprotein pp135 is an essential component of the nucleolus organizer region (NOR).

The association of phosphoproteins pp135 and pp105 with distinct substructures of the nucleolus was studied by cytochemical and immunological methods at the light microscopic and electron microscopic level. Both phosphoproteins exhibited a very high affinity for silver and Giemsa staining compared to other nucleolar proteins. Immunolocalization of pp135 and pp105 during mitosis by light microscopy revealed a tight association of pp135 with the silver staining nucleolus organizer region (NOR), whereas pp105 (cross-reacting with C23) appeared to be only partially associated with the NOR, exclusively at telophase. At the immunoelectron microscopic level the distribution of pp135 and pp105 was investigated in interphase nucleoli. Phosphoprotein pp135 was located in the fibrillar shell and pp105 in the fibrillar shell and the granular zone. The fibrillar centers were essentially free of both phosphoproteins..

Anaphase↗
PubMed1986

Characterisation of nucleolar proteins as autoantigens using human autoimmune sera.

Sera from 271 patients with systemic rheumatic diseases were screened for antinucleolar antibodies using immunofluorescence. Antinucleolar antibodies were found in the sera of 73% of patients with progressive systemic sclerosis, 4% of patients positive for autoantibodies with rheumatoid arthritis, and 8% of patients with rheumatoid arthritis associated with sicca complex, but not in patients with systemic lupus erythematosus, Sjögren's syndrome, undifferentiated connective tissue disease, or in healthy donors. The antinucleolar sera (n = 20) were analysed by immunoblotting techniques. In four sera antibodies against nucleolar proteins with molecular weights of 35 kD, 37 kD, 69 kD, 92-93 kD, and 93 kD could be immunodetected. The nucleolar autoantigens were extractable from the nucleoli together with the preribosomal particle fraction and could be solubilised by ribonuclease (RNAse) treatment. Their presence in the nucleolus was sensitive to actinomycin D treatment of cells. The 37 kD autoantigen could be identified as nucleolar phosphoprotein B23.

Animals↗
PubMed1985

Immunodetection of nitrocellulose-adhesive proteins at the nanogram level after trinitrophenyl modification.

A method for protein detection on nitrocellulose membranes based on modification with 2,4,6-trinitrobenzenesulfonic acid and reaction with anti-trinitrophenyl (TNP) serum as first antibody followed by peroxidase-conjugated second antibody is described. Protein quantities between 1 and 3 ng can be detected in the dot test. This method was used in a double immunodetection procedure after electrophoretic transfer of proteins localizing first a distinct antigen with its specific antiserum followed by visualization of the complete protein pattern on the same blot by the TNP/anti-TNP method as described above. As only water-soluble reagents are employed no shrinkage of the membrane occurs. Furthermore, the method can be used in a simultaneous immunodetection procedure visualizing the specific antigen together with TNP marker proteins using a mixture of the specific antiserum and the anti-TNP serum as first antibody.

Animals↗
PubMed1984

Isolation and localization of phosphoprotein pp 135 in the nucleoli of various cell lines.

Phosphoprotein pp 135 is one of the dominant proteins endogenously phosphorylated in cellular sonicates during short-time exposure to [gamma-32P]ATP. Mouse cells growing exponentially show the highest pp 135 level as determined by endogenous phosphorylation and immunobinding assays. Disruption of cells in the absence of calcium at low magnesium concentration renders more than 90% pp 135 into the cytosolic fraction. A five-step purification yields greater than 95% pure pp 135. The cellular location of pp 135 was determined with a rat anti-(mouse pp 135) serum by immunofluorescence in mouse cell lines and cryostat sections of normal mouse tissue. We observed fluorescence predominantly of nucleolar structures, confirmed by studies of isolated nuclei and nucleoli. Cross-reacting nucleolar phosphoproteins were identified in cell lines of other species with molecular masses of 128 kDa (human), 135 kDa (hamster) and 118 kDa (Drosophila). Endogenous phosphorylation of pp 135 investigated with purified mouse nucleoli showed optimal activity at isotonicity, pH 7.3, in the presence of 10 mM magnesium ions.

Animals↗
PubMed1983

Localization of phosphoprotein PP 105 in cell lines of various species.

The cellular location of phosphoprotein pp 105 was determined in various mouse cell lines with rabbit anti mouse pp 105 serum. Immunofluorescence was predominantly observed in the nucleoli in addition to a diffuse but weaker fluorescence of the whole nucleus. Cell surface fluorescence was obtained only with cells grown in suspension cultures. The presence of pp 105 in normal mouse tissue was demonstrated with tissue extracts by immunobinding assays. Cross-reacting phosphoproteins with the same molecular weight were detected in hamster and human cell lines as well as in chicken cartilage cells and Drosophila embryonic cells. Endogenous phosphorylation of pp 105 studied with purified mouse nucleoli showed optimal activity at isotonicity, pH 8.7, in the presence of 10 mM magnesium.

Animals↗
PubMed1983

Isolation and characterization of phosphoprotein pp 105 from simian virus 40-transformed mouse fibroblasts.

Investigation of the cellular distribution of a 105 kDa phosphoprotein (pp 105) in transformed mouse fibroblasts, showed that only a minor amount was located on the surface of logarithmically grown suspension cells. More than 90% of total pp 105 was contained in the cytosolic fraction representing about 0.2% of total cytosolic proteins. Surface and cytosolic pp 105 had identical phosphopeptide patterns. Cytosolic pp 105 was highly purified by ammonium sulfate precipitation followed by three chromatographic steps and gel electrophoresis. The purified pp 105 was capable of weak autophosphorylation. In the stationary growth phase of suspension cells, the amount of pp 105 detectable by endogenous phosphorylation was only 10-15% of that observed during logarithmic growth. pp 105 was also detected in normal mouse tissue and its distribution determined.

Animals↗
PubMed1981

Cell adhesion-dependent differences in endogenous protein phosphorylation on the surface of various cell lines.

Endogenous phosphorylation of intact cells was studied with four mouse, hamster and human cell lines using [gamma-32P]ATP and [gamma-32P]GTP as exogenous substrates. With all four cell lines distinct differences in the phosphoprotein patterns could be demonstrated for cells grown in suspension culture compared to cells grown in monolayers. Two major, apparently ubiquitous phosphoproteins with molecular weights of 135 000 (128 000 in HeLa cells) and 105 000, representing up to 60% of total phosphorylation, were phosphorylated only in cells grown in suspension. These phosphoproteins and the kinase(s) were located on the surface of the suspension cells. Evidence showed that phosphorylation was apparently not a true endogenous reaction, that rather it occurred by cell-cell collision, showing exponentially increasing 32P incorporation with increasing cell population density. Phosphorylation of pp135 and pp105 was established with ATP as well as with GTP and was not dependent on cyclic nucleotides cyclic AMP, cyclic GMP and cyclic CMP. The substrate-attached cells of all four cell lines have protein kinases on the cell surface. The lack of pp135 and pp105 phosphorylation may be due to the fact that these phosphoproteins are not expressed at all on the surface of substrate-attached cells or that these phosphoproteins are already fully phosphorylated.

Adenosine Triphosphate↗
PubMedDate not supplied

[Quantification of SS-B autoantibodies in patient sera using highly purified human SS-B antigens and their clinical interpretation].

For the identification of SS-B autoantibodies in the sera of patients with rheumatic diseases (n = 319) a sensitive and specific enzyme-linked immunosorbent assay was developed using highly purified human SS-B antigen as a reference antigen. In 30/319 patients' sera SS-B autoantibodies were detected. In this group, the ratio female : male patients was outstandingly high (29:1), the serum of the single male patient exhibiting the lowest titer of SS-B autoantibodies. The highest portion of anti-SS-B positive sera was observed in the group of patients with systemic Lupus erythematosus (48%) followed by the groups with undifferentiated connective tissue disease (17%), Sjögren-syndrome (13%) and rheumatoid arthritis (6%). The protein components reacting with human SS-B autoantibodies were characterized in permanent cell lines of various species. In each of 9 human cell lines only one component with a molecular weight of 49 kD reacted; in cell lines of other species analogously, only one component reacted, showing slightly different molecular weights depending on the species. The SS-B antigen isolated from human cells contained at least 7 isoelectric variants, all reacting with human SS-B autoantibodies.

Arthritis, Rheumatoid↗