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Biomedical subjects

R T Jones

Publications and source records attributed to R T Jones.

At least 127 records · Page 7Linked to original sources

Hemoglobin Brockton [beta 138 (H16) Ala----Pro]: an unstable variant near the C-terminus of the beta-subunits with normal oxygen-binding properties.

Hemoglobin Brockton [beta 138 (H16) Ala----Pro] is an unstable variant associated with a mild anemia. It has the same electrophoretic mobility as and cannot be resolved from Hb A. Oxygen affinity measurements of blood and hemolysate do not indicate biphasic oxygen saturation, showing that the functional properties of the variant are very similar to those of Hb A. This implies that the introduction of proline into the H-helix at position 138 does not disrupt the critical inter- and intrasubunit hydrogen bonds and salt bridges at the beta carboxyl-terminal dipeptide, since these polar interactions are essential for the normal oxygen-binding properties of hemoglobin. X-ray crystallographic data are consistent with these findings and show that the consequences of the beta 138 Ala----Pro substitution are almost entirely confined to the immediate vicinity of the mutation site. Instability probably results from the inability of a buried hydrogen bond to form between Pro 138 beta and Val 134 beta.

Amino Acid Sequence↗

Embryonic epsilon and gamma globin genes of a prosimian primate (Galago crassicaudatus). Nucleotide and amino acid sequences, developmental regulation and phylogenetic footprints.

Sequence analysis of epsilon and gamma genes and encoded globins and high-pressure liquid chromatography analysis of globin compositions in blood hemolysates obtained from embryos, fetuses and adults show that the prosimian primate Galago crassicaudatus expresses its epsilon and gamma genes only embryonically. Since rabbit, mouse and galago all have embryonic gamma genes but simian primates have fetal gamma genes, we conclude that gamma E evolved into gamma F in stem-simians. An elevated non-synonymous substitution rate characterizes this transition. The alignment of epsilon and gamma nucleotide sequences and the parsimoniously reconstructed evolutionary history of these sequences identify several anciently conserved cis-regulatory elements (phylogenetic footprints) important for gamma expression in primates and also cis-mutations which may have been involved in the recruitment of the gamma gene to a fetal program in simian primates.

Amino Acid Sequence↗

Affinity labeling of hemoglobin with 4,4'-diisothiocyanostilbene-2,2'-disulfonate: covalent cross-linking in the 2,3-diphosphoglycerate binding site.

The bifunctional reagent 4,4'-diisothiocyanostilbene-2,2'-disulfonate (DIDS) reacts with hemoglobin to give various products whose properties are dependent on the ligation state of the protein during the reaction. A major product obtained after reaction of (carbonmonoxy)hemoglobin with DIDS was a high oxygen affinity derivative [P50 = 1.4 mmHg, control P50 = 6 mmHg; 50 mM [bis(2-hydroxyethyl)-amino]tris(hydroxymethyl)methane (Bis-Tris), pH 7.4, 0.1 M Cl-, 25 degrees C] which contained two molecules of DIDS per tetramer resulting from adduct formation at each beta-chain amino terminus. In contrast, a major product of the reaction of deoxyhemoglobin with DIDS consisted of hemoglobin which had incorporated one molecule of DIDS per tetramer and was cross-linked between the beta-chain amino termini. This cross-linked hemoglobin was found to have a greatly decreased O2 affinity (P50 = 28 mmHg). Inhibition of the T to R transition due to the structural constraint produced by cross-linking the beta amino termini is likely to be a major factor in the decreased O2 affinity of this product. The structural and functional properties of this molecule make it a potential candidate for a cell-free blood substitute.

2,3-Diphosphoglycerate↗

The variant fetal hemoglobin F Texas I is abnormally acetylated.

Two related infants with indistinct slow-moving minor hemoglobin bands were detected by electrophoretic cord blood screening. The variant separated into a major and a minor band on isoelectric focusing (IEF), anion exchange chromatography (AEC), and high-performance liquid chromatography (HPLC). Analysis of the products of tryptic hydrolysis of the abnormal chain revealed a truncated A gamma T-1 peptide containing a glu----lys substitution at position 5, identifying the variant as Hb F Texas I. Microsequencing of the major early peak from AEC confirmed the substitution. However, gamma chains from the minor peak resisted Edman degradation and were shown to be acetylated by fast atom bombardment mass spectrometry (FABMS). Acetylation of HbF Texas I was nearly three times that of normal HbF measured in the same hemolysate, and this ratio remained constant for over 1 year in the proband. This represents the first demonstration of increased N alpha-acetylation of a variant fetal hemoglobin.

Acetylation↗

Long-term culture of hamster duodenal explants and cells.

Explants from adult Syrian hamster duodenum have been maintained in organ culture on gelatin sponges using CMRL 1066 serum supplemented media for 30 days. There was necrosis of the tall villi architecture during the 1st week in culture while columnar and mucous cells survived, migrated, and replicated along portions of the explant basement membrane and in the gelatin sponge matrix. Cells in the sponge multiplied and formed epithelial sheets which showed villus projections and cyst configurations. The cells in these epithelial structures were attached to one another by junctional complexes. Epithelial cells were isolated from the sponge matrix by collagenase digestion and were successfully grown in culture. These duodenal explants and cells show potential for use as in vitro models for experimental studies, involving acute and chronic response of cells to injury or carcinogen-induced transformation.

Animals↗

A stoma jig for surgical craft workshops.

A new jig simulating the abdominal cavity and wall is described. The techniques of defunctioning colostomy and closure of colostomy, end colostomy and ileostomy can be performed. If required the techniques of mass or layered closure of an abdominal incision can also be practised.

Abdomen↗

Combined atropine and 2-PAM Cl effects on tracking performance and visual, physiological, and psychological functions.

Combinations of atropine (up to 4 mg.70 kg-1) and 2-PAM Cl (up to 1200 mg.70 kg-1) were studied for their effects on a pursuit tracking task, six visual functions, heart rate and blood pressure, and cognitive functions as measured by six psychological tests. Tracking performance in both bright light and dim light was significantly degraded up to 3.5 h after injection. High- and low-contrast near acuity was significantly altered up to 6 h after injection, whereas accommodation and pupil size remained altered for 24 h. Elevated pulse rates were observed for 4 h. Elevated systolic blood pressures were observed for 2 h while diastolic pressures remained elevated for 6 h. No drug effects were found for the psychological tests. Overall, the observed effects of these two drugs in combination are qualitatively similar to those of atropine alone although they are of greater magnitude. On two measures (accommodation and diastolic blood pressure), 2-PAM Cl was found to significantly potentiate the atropine effect.

Accommodation, Ocular↗

Acute effects of cocaine on catecholamines and cardiac electrophysiology in the conscious dog.

To evaluate the electrophysiologic and neurohumoral effects of cocaine on the sinus node, atrium, atrioventricular node, and His-Purkinje system, 14 conscious dogs were studied before and after an infusion titrated to maximum tolerance or to increase the systolic blood pressure by at least 15%. Plasma cocaine and catecholamine levels, blood pressure, surface and intracardiac electrograms were recorded. Programmed electrical stimulation was performed from the right atrium. The sinoatrial conduction time, paced PR interval, atrioventricular nodal conduction time, atrioventricular nodal effective refractory period and blood pressure increased. Plasma noradrenaline, adrenaline and dopamine increased and remained at two to three times control levels throughout the study although plasma cocaine levels declined. Adrenaline levels were strongly correlated with the hemodynamic response while plasma cocaine levels were not. No sustained spontaneous atrial or ventricular arrhythmias were recorded after cocaine infusion. Atrial fibrillation with a slow ventricular response was induced by pacing after cocaine in only three dogs. There were no significant changes in pacing threshold, right intra-atrial conduction time, infranodal conduction time, heart rate, QRS or QT. No pacing induced infranodal block occurred. The cocaine dose rate infused was 2.8 +/- 1.2 mg/kg; cocaine plasma levels were 1402 +/- 885 ng/mL immediately after the initial infusion and 525 +/- 321 ng/mL at the end of the study. It is concluded that in normal canine heart cocaine increases blood pressure, sinoatrial conduction time, atrioventricular nodal effective refractory period, paced PR interval, atrioventricular nodal conduction time, plasma catecholamines and susceptibility to atrial fibrillation. These findings cannot be explained solely by increased sympathetic nervous system activity and suggest participation of the parasympathetic nervous system.

Animals↗

Determination of benzoylecgonine and cocaine in biologic fluids by automated gas chromatography.

A method is described for extraction of the cocaine metabolite benzoylecgonine, conversion to the butyl ester derivative and gas chromatographic analysis using packed or capillary columns. Using a capillary column, cocaine and benzoylecgonine may be determined simultaneously. The extraction scheme has been designed to facilitate processing of large numbers of samples generated in pharmacokinetic studies. Structural analogues, m-toluylecgonine and m-toluylecgonine methyl ester, are used as internal standards. Concentrations as low as 10 ng/ml in 1-ml samples of plasma or urine are readily determined. Between-run coefficients of variation were 1.01% for cocaine and 4.18% for benzoylecgonine for concentrations of 75 and 350 ng/ml, respectively.

Autoanalysis↗

Cocaine-calcium channel antagonist interactions.

Diltiazem, a benzothiazepine calcium channel antagonist, was given to six healthy men as a single 60 mg oral dose 120 min before IV injection of cocaine (0.2 mg/kg) in a double-blind, placebo-controlled, two-session study. Diltiazem alone produced no significant effects. Cocaine increased blood pressure, heart rate, pupil size and subjective "high" ratings, and decreased skin temperature. Diltiazem pretreatment diminished the cocaine effect on skin temperature, but did not otherwise alter the response to cocaine. Calcium channel antagonists diminish the effects of cocaine in vitro and in animals. Dosage considerations may be critical because of the differential sensitivity of various tissues to calcium channel antagonists.

Adult↗

Increased susceptibility to memory intrusions and the Stroop interference effect during acute marijuana intoxication.

The marijuana-induced acute memory impairment was assessed in a double-blind, crossover experiment. Twelve males smoked NIDA-supplied cigarettes containing 1.2% delta-9-tetrahydrocannabinol (THC) or cannabinoid-exhausted marijuana (placebo) in counterbalanced order on 2 days 1-3 weeks apart. Practice, pre- and postsmoking test sessions were conducted with the Paced Auditory Serial Addition Test, Stroop Color and Word Test, and alternate forms of the Randt Memory Battery and the Controlled Oral Word Association Test. A significantly greater number of short story omissions and intrusions occurred in delayed free recall after marijuana. Immediate and sustained attention, controlled retrieval from semantic memory, and speed of reading and naming colors were not affected. The Stroop interference effect was significantly greater following marijuana. Subjects appeared to experience parallel difficulties in inhibiting associations to the new material and inhibiting the overlearned response of reading in a new learning task. Marijuana may compromise associative control, presumably a cognitive process inherent in memory function.

Adult↗

Hamster tracheal organ culture in serum-free media: a quantitative comparison of in vitro epithelial morphology with that of in vivo controls.

The epithelial morphology of the hamster trachea in serum-free organ culture was compared with that of age-matched in vivo control tissues by collecting and statistically analyzing several quantifiable parameters. By this technique it was possible to detect both subtle and dramatic epithelial alterations. Midtracheal tissues from 6-wk-old male Syrian golden hamsters were used as the explants. Explants were placed on Gelfoam sponges and cultured for 1,2, and 3 wk in CMRL 1066 alone and in CMRL 1066 to which seven factors were added: insulin and transferrin (5 micrograms/ml); hydrocortisone (5 X 10(-7) M); epidermal growth factor (5 ng/ml); bovine pituitary extract (0.5%); and phosphoethanolamine and ethanolamine (5 X 10(-5) M). The following data were collected and statistically analyzed for each tracheal ring: number of epithelial cells; proportion and number of each cell type; basement membrane length; linear density of epithelial cells; epithelial height; and mitotic index. Compared to controls, ciliated cells decreased by 52% during washes in Leibovitz (L15) medium and tissue manipulation performed before culture and this loss persisted after culture for 1 wk. Explants cultured without the factors showed marked changes after 2 and 3 wk including epithelial thickening and folding, which was associated with increased linear density. Many cells in these specimens could not be categorized by type (22% were unidentifiable after 3 wk). Epithelial migration onto the outside of the explant was inhibited. In contrast, explants cultured with the factors maintained a morphology similar to controls at 2 and 3 wk and epithelial migration onto the outside of the explant was supported. This study shows that explants in CMRL 1066 with the seven factors provide a useful biological model for the in vitro study of mucociliary respiratory epithelium.

Animals↗

Cocaine effects on electroencephalographic cognitive event-related potentials and performance.

The effects of cocaine on electroencephalographic indices, information processing and motor performance were measured in 12 normal volunteers. They were tested with an auditory continuous performance task before, 60 and 180 min after a 3 mg/kg oral dose of cocaine hydrochloride or placebo in a double-blind counterbalanced crossover design. EEG from Fz, Cz and Pz were averaged for the eye movement-free correct trials. N100, P200, P300 components and CNV were measured. Cocaine increased N100 amplitude for the warning stimulus and CNV amplitude. P300 amplitude and latency, reaction time and sensory sensitivity (d') were not altered by cocaine. The increase in N100 and CNV amplitude may reflect mobilization of additional attentional resources to produce optimal performance on the task when subjects were distracted by the cardiovascular, subjective and other cocaine effects.

Adult↗

Speech event related potentials reflect linguistic content and processing level.

Amplitude changes in the auditory event related response reflect differences in linguistic content and the level of processing of spoken sentences. Thus, the neurophysiology of the cognitive processes underlying speech perception can be evaluated by noninvasive techniques. Semantically correct, semantically incorrect, and grammatically incorrect sentences were processed either semantically or syntactically by 25 subjects while their EEGs were recorded. The amplitude of the P250 component to each word varied with processing level. A slow positive wave appeared after semantically correct sentences regardless of the linguistic processing level. N480 and P780 components were observed following words which made the sentences incorrect.

Adult↗

Effect of maternal vaccination on the susceptibility of growing pigs to leptospiral infection.

Serum samples were collected from 30 piglets, derived from 17 litters, whose dams had been vaccinated against leptospirosis. Microscopic agglutination test (MAT) titres against Leptospira interrogans serovar pomona varied greatly from pig to pig; there was less variation among littermates. Titres declined between 4 and 10 weeks of age, with an uncorrected half-life of 15.5 days, consistent with IgG being the main antibody class involved. Twelve pigs, 4 derived from unvaccinated sows and 8 from sows vaccinated against leptospirosis, were challenged intravenously at 8 weeks of age with leptospires of serovar pomona. Colostrum-derived antibody protected 4 out of 8 pigs, and in 1 of the remaining 4 the serological response was reduced. Three of the protected pigs showed reduced serological responses and in the fourth the response was strong, but delayed. All of the pigs derived from unvaccinated sows developed leptospiraemia and leptospiruria and showed strong serological responses. Protection by colostrum-derived antibody bore an inexact relationship to MAT titre, but a titre of 16 appeared to be sufficient for protection.

Agglutination Tests↗