Nerve growth factor alpha subunit: effect of site-directed mutations on catalytic activity and 7S NGF complex formation.

Mouse alpha- and gamma-nerve growth factor (NGF) are glandular kallikreins that form a non-covalent complex (7S NGF) with beta-NGF. gamma-NGF is an active arginine-specific esteropeptidase; the alpha-subunit is catalytically inactive and has a zymogen-like conformation. Site-directed mutagenesis of alpha-NGF to alter the N-terminus and three residues in loop 7, a region that contributes to the catalytic center, restored substantial catalytic activity against N-benzoyl arginine-p-nitroanilide as substrate in two derivatives although they were not as active as recombinant gamma-NGF.

Modern pollen-representation of some boreal species on islands in a large lake in Canada.

Studies of pollen source areas of closed-canopy sites are contradictory. Some authors found that closed-canopy sites mainly collect local pollen while others found more distant sources. This dichotomy might stem from the use of canopies of varying degrees of closure, and from variations in the pollen productivity of the local vegetation and the background pollen rain.

Activation of the Stat3 signaling pathway is required for differentiation by interleukin-6 in PC12-E2 cells.

The role of signal transducer and activator of transcription (STAT) signaling pathways in the interleukin-6 (IL-6)-induced morphological differentiation of PC12-E2 cells was assessed using wild type and dominant negative mutants of Stat1 and Stat3, containing Tyr --> Phe (YF), Ser --> Ala (SA), and the double mutations (DM), respectively. FS3-YF or FS3-DM markedly inhibited the IL-6-induced response, but overexpression of FS3-SA caused only a modest inhibition. Expression of all Stat3 mutants had no effect on NGF-induced neurite outgrowth.

Effectiveness of an information broker service.

The many disparate databases existing within the same health care organization create confusion and frustration for the consumer trying to get integrated information. The purpose of this research was to create a regional service that would provide a customer oriented information broker service with a single point of contact and guaranteed performance. From 1/98-6/99 there were 34 requests made.

Efficacy of computerized decision support for mechanical ventilation: results of a prospective multi-center randomized trial.

200 adult respiratory distress syndrome patients were included in a prospective multicenter randomized trial to determine the efficacy of computerized decision support. The study was done in 10 medical centers across the United States. There was no significant difference in survival between the two treatment groups (mean 2 = 0.

Comparison of the intracellular signaling responses by three chimeric fibroblast growth factor receptors in PC12 cells.

Stably transfected PC12 cell lines expressing similar amounts of chimeric receptors composed of the extracellular domain of the human platelet-derived growth factor (PDGF)beta receptor and the transmembrane and intracellular domains of the fibroblast growth factor receptors (FGFRs) 1, 3, and 4 undergo ligand-induced differentiation. The FGFR1 chimera (PFR1) is the most potent of the three, and PFR4 requires more frequent (every 24 hr) addition of ligand to maintain the response. Both PFR1 and -3 also show significant ligand-independent autophosphorylation but PFR4 does not.

Yeast methionine aminopeptidase I. Alteration of substrate specificity by site-directed mutagenesis.

In eukaryotes, two isozymes (I and II) of methionine aminopeptidase (MetAP) catalyze the removal of the initiator methionine if the penultimate residue has a small radius of gyration (glycine, alanine, serine, threonine, proline, valine, and cysteine). Using site-directed mutagenesis, recombinant yeast MetAP I derivatives that are able to cleave N-terminal methionine from substrates that have larger penultimate residues have been expressed. A Met to Ala change at 329 (Met206 in Escherichia coli enzyme) produces an average catalytic efficiency 1.

Yeast (Saccharomyces cerevisiae) methionine aminopeptidase I: rapid purification and improved activity assay.

Methionine aminopeptidase (MetAP), in two isoenzymic forms, is responsible for hydrolysis of the initiator methionine residues from the majority of newly synthesized proteins. It is an essential gene product and is ubiquitously found in archaea, eubacteria, and lower and higher eukaryotes. MetAP also has a potentially important role in the production of recombinant proteins, since failure to correctly remove initiator methionine residues can result in a product that is inactive or immunogenic.

Automated key process monitors for patient care documentation.

Complete, accurate and timely patient care documentation is an essential part of the practice of medicine. As with any other process in medicine or industry, continuous quality improvement (CQI) is essential to assure the highest quality at the lowest cost. CQI requires objective key process measures that can be assessed routinely.

Spouse abuse: physician guidelines to identification, diagnosis, and management in the uniformed services.

Spouse abuse is a major social issue in our country. Based on national samples, between 10 and 20% of couples report some type of violence during the course of their marriage. In the United States, 1.

Yeast methionine aminopeptidase I can utilize either Zn2+ or Co2+ as a cofactor: a case of mistaken identity?

Yeast methionine aminopeptidase I (MetAP I) is one of two enzymes in Saccharomyces cerevisiae that is responsible for cotranslational cleavage of initiator methionines. It has previously been classified as a Co2+ metalloprotease in all prokaryotic and eukaryotic forms studied. However, treatment of recombinant apo-MetAP I with 12.

Effect of transmembrane and kinase domain mutations on fibroblast growth factor receptor 3 chimera signaling in PC12 cells. A model for the control of receptor tyrosine kinase activation.

The effect of six point mutations causing various human skeletal dysplasias, occurring in the transmembrane (TM) and kinase domains (KD) of fibroblast growth factor receptor 3, were introduced into a chimera composed of the extracellular domain of human platelet-derived growth factor beta and the TM and intracellular domains of hFGFR3. Stable transfectants in rat PC12 cells showed distinct differences in the two classes of mutations. The cells containing TM mutants displayed normal expression and activation but higher responsiveness to lower doses of ligand.

FGF signal transduction in PC12 cells: comparison of the responses induced by endogenous and chimeric receptors.

Rat phaeochromocytoma (PC12) cells respond to many growth factors and produce different phenotypes, including neurite outgrowth. Receptor tyrosine kinases (RTK), which activate multiple signalling pathways in response to ligand binding, initiate many of these. One such family of receptors, the fibroblast growth factor receptor (FGFR), has four different members and expresses at least three of these in PC12 cells.

Effects of simulated lairage conditions on the physiology and behaviour of pigs.

The responses of pigs to being driven and mixed together in lairage were investigated. Five groups of six and five groups of seven 70 kg pigs were transported for 40 minutes on a lorry and then subjected to one of the following treatments: two groups were driven down a passage; four groups (A to D) were mixed together for one hour (A and B together, and C and D together); and, four groups were driven down the passage and then mixed ('combined treatment'). The pigs' behaviour was recorded, skin damage was scored and saliva samples were taken for analysis of cortisol.

Effects of lavender straw on stress and travel sickness in pigs.

Objective: To observe pigs during road journeys in order to establish whether lavender straw was likely to decrease stress and incidence of travel sickness.

Subjects: Forty 70-kg Large White pigs were transported by road for 2 hours, 20 animals each day, over a 2-day period.

Design: On day 1, ample wheat straw was provided as bedding such that the floor of the vehicle was entirely covered (straw condition).

N-terminal processing: the methionine aminopeptidase and N alpha-acetyl transferase families.

Removal of the initiator methionine and/or acetylation of the alpha-amino group are among the earliest possible chemical modifications that occur during protein synthesis in eukaryotes. These events are catalyzed by methionine aminopeptidase and N alpha-acetyltransferase, respectively. Recent advances in the isolation and characterization of these enzymes indicate that they exist as isoforms that vary in cellular location, function, and evolutionary origins.

Effects of transport and indomethacin on telemetered body temperature and release of cortisol and prolactin in pre-pubertal pigs.

Previous research indicates that acute physical stress (restraint) raises core temperature in growing pigs via a prostaglandin-dependent mechanism. This study investigated whether transport stress affects body temperature in pigs and whether any such changes might involve endogenous prostaglandins. Pre-pubertal boars (n = 7) were implanted with venous catheters and biotelemetry devices for the measurement of core temperature.

Applications of site-specific chemical modification in the manufacture of biopharmaceuticals: I. An overview.

The chemical modification of proteins has long been a useful approach to elucidating structure-function relationships. Recently, these same chemistries have been finding application in the preparation of proteins for both diagnostic and therapeutic use. These applications include alterations to introduce new properties, such as improved stability, or new traits, such as drug binding and transport, that take advantage of both broad and narrow ranges of selectivity.

Telling rights from wrong.

Is it right to medicate patients without their knowledge and consent? Or to tell an adult woman what she can eat? The UKCC is producing new guidance specifically to help mental health and learning disabilities nurses with the daily dilemmas of clinical practice.

Differential utilization of ShcA tyrosine residues and functional domains in the transduction of epidermal growth factor-induced mitogen-activated protein kinase activation in 293T cells and nerve growth factor-induced neurite outgrowth in PC12 cells. Identification of a new Grb2.Sos1 binding site.

By transient expression of both truncated forms of p52(SHCA) and those with point mutations in 293T cells, it has been shown that, in addition to Tyr-317, Tyr-239/240 is a major site of phosphorylation that serves as a docking site for Grb2.Sos1 complexes. In addition, analysis of epidermal growth factor (EGF)-induced activation of mitogen-activated protein kinase in 293T cells showed that the overexpression Shc SH2 or phosphotyrosine binding (PTB) domains of ShcA alone has a more potent negative effect than the overexpression of the forms of ShcA lacking Tyr-317 or Tyr 239/240 or both.