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Biomedical subjects

S C Hall

Publications and source records attributed to S C Hall.

At least 19 recordsLinked to original sources

Rapid mass spectrometric peptide sequencing and mass matching for characterization of human melanoma proteins isolated by two-dimensional PAGE.

We report a general mass spectrometric approach for the rapid identification and characterization of proteins isolated by preparative two-dimensional polyacrylamide gel electrophoresis. This method possesses the inherent power to detect and structurally characterize covalent modifications. Absolute sensitivities of matrix-assisted laser desorption ionization and high-energy collision-induced dissociation tandem mass spectrometry are exploited to determine the mass and sequence of subpicomole sample quantities of tryptic peptides. These data permit mass matching and sequence homology searching of computerized peptide mass and protein sequence data bases for known proteins and design of oligonucleotide probes for cloning unknown proteins. We have identified 11 proteins in lysates of human A375 melanoma cells, including: alpha-enolase, cytokeratin, stathmin, protein disulfide isomerase, tropomyosin, Cu/Zn superoxide dismutase, nucleoside diphosphate kinase A, galaptin, and triosephosphate isomerase. We have characterized several posttranslational modifications and chemical modifications that may result from electrophoresis or subsequent sample processing steps. Detection of comigrating and covalently modified proteins illustrates the necessity of peptide sequencing and the advantages of tandem mass spectrometry to reliably and unambiguously establish the identity of each protein. This technology paves the way for studies of cell-type dependent gene expression and studies of large suites of cellular proteins with unprecedented speed and rigor to provide information complementary to the ongoing Human Genome Project.

Amino Acid Sequence

Sodium nitroprusside metabolism in children during hypothermic cardiopulmonary bypass.

Ten children, aged 1-7 yr, presenting for repair of complex congenital heart lesions, were prospectively studied. A ketamine, halothane/isoflurane, and fentanyl anesthetic was used. After initiation of hypothermic cardiopulmonary bypass, sodium nitroprusside (SNP) was titrated as necessary to maintain a target mean arterial blood pressure of 35-60 mm Hg. Blood samples drawn immediately prior to starting SNP infusion, every 15 min during infusion, and at 1, 4, and 24 h postinfusion were analyzed for whole blood cyanide (CN-) and serum thiocyanate (SCN-). Blood gas analysis was performed every 30 min during SNP infusion. A maximum CN- level > or = 1.0 micrograms/mL was observed in two children; four others had maximum CN- levels between 0.5 micrograms/mL and 1.0 micrograms/mL (normal, < 0.2 micrograms/mL). No child had a clinically important increase of SCN- subsequent to SNP infusion. There was substantial variability in observed CN- accumulation during SNP infusion. CN- levels during the first 60 min correlated with the average SNP rate of administration (P = 0.02). Cyanide levels rapidly decreased after termination of SNP infusion and were undetectable 4 h postinfusion. Despite the short-term increase of CN- level, no child showed biochemical signs of toxicity (acidosis or increased mixed venous oxygen tension).

Cardiopulmonary Bypass

Comparison of a rigid laryngoscope with the ultrathin fibreoptic laryngoscope for tracheal intubation in infants.

The flexible ultrathin fibreoptic laryngoscope allows placement of endotracheal tubes as small as 2.5 mm internal diameter. The purpose of this study was to document the safety and efficacy of intubation using an ultrathin fibreoptic laryngoscope. Proved safety and efficacy would justify the routine use of fibreoptic laryngoscopy in normal infants to maintain skills needed for management of the difficult infant airway. In this prospective study, 40 infants < 24 mo of age scheduled for elective surgery were randomly divided into two equal groups. After inhalation induction of anaesthesia, in 20 infants the trachea was intubated using direct rigid laryngoscopy, and in 20 using the ultrathin fibreoptic laryngoscope (size 1.8 mm OD) Olympus LFP. Time to successful intubation was recorded, as well as blood pressure, heart rate, end-tidal CO2 and oxygen saturation. Airway trauma in the operating room, the post-anaesthesia care unit, and on the first postoperative day was recorded. The intubation times using rigid laryngoscopy were less than those using fibreoptic laryngoscopy (13.6 +/- 0.9 sec (mean +/- SEM) vs 22.8 +/- 1.7 sec; P < 0.01). Oxygen saturation and end-tidal CO2 readings were not different between the two groups. After intubation, blood pressure and heart rate increased equally in both groups, returning to normal within one to two minutes. There was no difference in the airway trauma between groups. We conclude that the ultrathin fibreoptic laryngoscope is a safe and effective method for tracheal intubation in infants and may be used routinely in order to maintain fibreoptic airway skills.

Anesthesia, Inhalation

Anesthetic considerations for the pediatric plastic surgical patient.

An understanding of the anesthesiologist's concerns during pediatric plastic surgical procedures can facilitate the coordination of efforts between the multiple services involved in the care of these children. Prior to surgery, the child's history is obtained and physical examination is performed. The condition of the airway is the primary concern. Preoperative medication is usually given by oral routes to avoid injections. Induction intubation, maintenance, and emergence are discussed. Specific postanesthesia care is described. Special precautions are given for children having surgical repair of craniofacial anomalies. Cleft lip and palate and mandibular advancement are described.

Age Factors

Specificity of DNA recognition in the nucleoprotein complex for site-specific recombination by Tn21 resolvase.

Resolvases from Tn3-like transposons catalyse site-specific recombination at res sites. Each res site has 3 binding sites for resolvase, I, II, and III. The res sites in Tn3 and Tn21 have similar structures at I and II but they differ at III. Mutagenesis of the Tn21 res site showed that sub-site III is essential for recombination though the sequences in III that are recognized by Tn21 resolvase are positioned differently from the equivalent sequences in the Tn3 site. The deletion of III caused a 1,000-fold drop in the rate of recombination. But other mutations at III, changing 3 or 4 consecutive base pairs, caused only 1.5- to 4-fold decreases in rate, even when the mutations were in target sequences for this helix-turn-helix protein. The reason why Tn21 resolvase has similar activities at a number of different DNA sequences may be due to the multiplicity of protein-protein and protein-DNA interactions in its recombinogenic complex. This lack of precision may be a general feature of nucleoprotein complexes.

Base Sequence

Mass spectrometric and Edman sequencing of lipocortin I isolated by two-dimensional SDS/PAGE of human melanoma lysates.

We have integrated preparative two-dimensional polyacrylamide gel electrophoresis with high-performance tandem mass spectrometry and Edman degradation. By using this approach, we have isolated and identified, by partial sequencing, a human melanoma protein (34 kDa, pI 6.4) as lipocortin I. To our knowledge, this protein was not previously known to be associated with melanoma cells. The identity of the protein was confirmed by two-dimensional immunoblot analysis. High-energy collision-induced dissociation analysis revealed the sequence and acetylation of the N-terminal tryptic peptide and an acrylamide-modified cysteine in another tryptic peptide. Thus, knowledge concerning both the primary structure and covalent modifications of proteins isolated from two-dimensional gels can be obtained directly by this approach, which is applicable to a broad range of biological problems.

Amino Acid Sequence

Direct comparison between protons and alpha-particles of the same LET: I. Irradiation methods and inactivation of asynchronous V79, HeLa and C3H 10T1/2 cells.

A direct comparison was carried out of the biological effectiveness of protons and alpha-particles of the same linear energy transfer (LET) under identical conditions with a variety of in vitro biological systems. Monolayers of mammalian cells were irradiated with accelerated beams of protons (1.2 and 1.4 MeV) and alpha-particles (30 and 35 MeV) corresponding to LETs of 23 and 20 keV microns-1 for each particle type. For V79-4 cells it was observed that the linear term of the dose-response for cell inactivation by protons was significantly greater than that for alpha-particles of the same LET. For HeLa and HeLa S3 cells, also, the linear term appeared to be greater for protons, but this was not observed with more limited data for C3H 10T1/2 cells. The result for V79 cells is in agreement with the report of Belli et al. (1989) who observed that the biological effectiveness of protons rose sharply between 17 and 30 keV microns-1 in strong contrast to alpha-particles which reached a peak effectiveness at greater than 100 keV microns-1. These results place new constraints on the biologically relevant features of the microscopic structure of radiation tracks, and have implications for the mechanistic and practical comparison between radiations.

Alpha Particles

Anaesthetic management of Miller's syndrome.

Miller's syndrome is a rare congenital disorder with facial features similar to that of Treacher-Collins syndrome. This report details the anaesthetic management of an infant during multiple surgical procedures, beginning with pylormyotomy at one month of age. Airway management was difficult because of severe micrognathia and was accomplished using an awake intubation with a conventional straight blade modified for continuous administration of oxygen ("oxyscope"). Due to recurrent upper airway obstruction and the anticipated need for multiple surgical procedures in the first years of life, a tracheostomy was placed. Because of the multiple airway, orthopaedic, and nutritional difficulties, it is important that a prospective, multidisciplinary approach be used in these patients' care. Consideration should be given to early tracheostomy for airway maintenance.

Anesthesia, Inhalation