Characterization of neurotensin binding to rat gastric smooth muscle receptor sites

The binding of monoiodo 125I-Trp11-neurotensin to purified rat gastric fundus smooth muscle plasma membranes was characterized. Specific binding of ligand in subcellular fractions from rat fundus smooth muscle showed a distribution that paralleled that of several plasma membrane marker enzymes. 125I-Trp11-neurotensin binding to smooth muscle plasma membranes at 25 degrees C was maximal at 30 min, reversible and saturable. Scatchard analysis of equilibrium data indicated the existence of two classes of binding sites with dissociation constants (Kd) of 56 pmol and 1.92 nM, and corresponding binding capacities (Bmax) of 6.6 fmol/mg and 11.4 fmol/mg of membrane protein. Analogues and fragments of neurotensin competed for 125I-Trp11-neurotensin binding with a rank order of potency similar to that previously reported for their contracting effect in rat fundus strips. Na+ decreased in a concentration dependent manner the binding of labelled ligand to the high affinity site. At 100 mM, Na+ induced a 6-fold increase in the IC50 of neurotensin for inhibition of 125I-Trp11-neurotensin binding. At this concentration of Na+, the IC50 for neurotensin was 1 nM, a value close to the Kd of the low affinity site.     

Improved techniques for use of the triploid cell marker in the axolotl, Ambystoma mexicanum

Techniques for using the triploid cell marker for studying cell lineage during the development and regeneration of the axolotl limb are described. Triploid animals possess cells with three nucleoli while diploid animals possess cells with two nucleoli. We have developed a technique for isolating the limb dermis as a sheet of cells for whole-mount analysis of cellular ploidy. Whole-mount tissue preparations as well as paraffin-embedded sectioned tissues were stained specifically for nucleoli with bismuth. Cell counts from a number of triploid and diploid dermal preparations show that (1) diploid dermal cells never possess three nucleoli, (2) the frequency of trinucleolate cells in whole-mount triploid dermal preparations is not 100% but varies between animals from 30 to 76%, (3) within a single triploid animal, the frequency of trinucleolate cells in different dermal preparations is constant. These data establish the usefulness of this technique and emphasize the need for appropriate control cell counts when using the triploid cell marker in the axolotl.     

Chromosome studies in 496 infertile males with a sperm count below 10 million/ml

Summary Chromosome studies were performed on 106 men with azoospermia and 390 men with oligozoospermia (consistant sperm count below 10 million/ml). Constitutional chromosome abnormalities were found in 14.1% of the azoospermia group and in 5.1% of the oligozoospermia group. An overall incidence of 7.1% constitutional abnormalities indicates that this criterion of selection may be advisable for routine chromosome analysis of infertile men. A reduction of 25% in the workload increases the yield of chromosome abnormalities in the group of infertile men to 10–14 times above that expected in the normal population.     

Head injuries related to the use of baby walkers.

To determine what proportion of head injuries in children under 24 months of age who presented to an emergency department were related to the use of baby walkers, we reviewed the charts of 52 such children. Walkers were involved in 42% of the head injuries in the children under 12 months of age and in none of those in the children aged 12 to 24 months. All walker-related injuries, including skull fractures in three children, involved stairs (p less than 0.001). Questionnaires were also sent to all families with children aged 3 to 18 months attending a private pediatric practice to determine the prevalence of falls involving baby walkers among these children and the factors associated with such falls. Of the 152 responding families 82% reported using or having used a walker. Thirty-six percent of the families reported that their child had a fall while in a walker, with 8.8% of the falls resulting in contact with a doctor. Walker-related falls were directly associated with time spent in the walker (p less than 0.001) and with a previous fall from the walker by an older sibling (p less than 0.03). Since there is no demonstrated benefit of walkers, their use should not be encouraged, and parents should be advised of their potential danger.     

Precordial exercise mapping: improved diagnosis of coronary artery disease.

Surface mapping of the exercise electrocardiogram (ECG) provides a measure of the precordial area, severity, and time course of ST-segment changes occurring after exercise. Sixteen-lead isopotential surface maps were recorded before and after exercise in 109 patients with probable angina who subsequently underwent coronary arteriography. In addition, exercise ECGs with three orthogonal leads were obtained in 53 of these patients, and with a single unipolar chest lead in all 109. Of the 109 patients, 85 had significant (greater than or equal to 70%) narrowing of at least one major coronary artery. The sensitivities of the precordial surface mapping, orthogonal leads, and single chest lead (V5) when compared with the findings at coronary arteriography were 95%, 68%, and 64% respectively, while the specificities of the three lead systems did not differ significantly. The technique of precordial surface mapping after exercise may easily be applied in clinical practice and requires only conventional equipment available in most hospitals. It effectively aids diagnosis of coronary artery disease.     

Conformational analysis of a snake neurotoxin by prediction from sequence, circular dichroism, and raman spectroscopy.

The secondary structure of the major neurotoxin from the sea snake Lapemis hardwickii was investigated by several methods of conformational analysis: structure prediction, circular dichroism, and laser Raman spectroscopy. From the primary structure, secondary structure prediction yielded two regions of β-sheet structure at residues 1–7 and 41–45. β-Turns were predicted at residues 14–17, 20–23, 30–33, 37–40, and 46–49. From the predictions, the toxin appears to be composed of approximately 20% β-sheet and 33% β-turn. The CD spectrum of the native toxin appears to be a hybrid of model spectra for β-sheet and β-turn proteins. The pH perturbation studies on the toxin observed by CD demonstrated that the toxin is a very stable molecule except at extremely high or low pH values. The Raman data indicated that the toxin contains both antiparallel β-sheet and β-turn structure. Using two methods of secondary structure quantitation from Raman spectra the molecule was calculated to contain 35% β-sheet from one method and 27% from the other. Overall, the various methods demonstrate that the toxin is composed of β-sheet and β-turn structure with little or no α-helix present. From the comparison of these different techniques appreciation can be gained for the necessity of several methods when identifying and quantitating secondary structure.