Cloning, purification, and preliminary characterization by circular dichroism and NMR of a carboxyl-terminal domain of the bacteriophage P22 scaffolding protein

Assembly of double-stranded DNA viruses and bacteriophages involves the polymerization of several hundred molecules of coat protein, directed by an internal scaffolding protein. A 163-amino acid carboxyl-terminal fragment of the 303-amino acid bacteriophage P22 scaffolding protein was cloned, overexpressed, and purified. This fragment is active in procapsid assembly reactions in vitro. The circular dichroism spectrum of the fragment, as well as the 1D-NMR and 15N-1H HSQC spectra of the uniformly-labeled protein, indicate that stable secondary structure elements are present. Determination of the three dimensional packing of these elements into the folded scaffolding protein fragment is underway. Structure-based drug design targeted at structural proteins required for viral assembly may have potential as a therapeutic strategy.     

The use of Cordanum in combination with Corinfar-Retard for the treatment of ectopic arrhythmias in the sick sinus syndrome in patients with ischemic heart disease

38 patients with ischemic heart disease (IHD) and sick sinus syndrome (SSS) received combined therapy with nifedipine (Corinfar-Retard) and talinolol (Cordanum). The former drug had a positive chronotropic effect on the heart, the latter's chronotropic effect was slightly negative. All the patients had sinus bradycardia and ectopic arrhythmia which needed therapeutic correction: supraventricular and ventricular extrasystoles, fibrillation paroxysms or/and atrial flutter, paroxysmal supraventricular tachycardia, ventricular tachycardia. Cordanum was given in a dose 50 mg twice a day, Corinfar-Retard 20 mg twice a day for 16 days. 30 patients responded to the treatment. In addition to good subjective response, episodes of extrasystoles, paroxysms, flutter and fibrillation occurred much less frequently. Side effects resulted in the treatment discontinuation in 3 patients.     

Binding of thyroid hormone to the goat testicular Leydig cell induces the generation of a proteinaceous factor which stimulates androgen release

Chromosome 3 comprises 7% of the genome and contains at least 210 Mb of DNA. To expedite the analysis of this chromosome, we have assembled a somatic cell hybrid mapping panel that subdivides human chromosome 3 into 23 intervals using a total of 19 hybrids. Hybrids were constructed from 16 patients' cells containing chromosome 3 translocations. All of these hybrids selectively retained the derivative 3 chromosome. In addition, we utilized 2 radiation-reduced hybrids and 3 hybrids carrying spontaneous translocations between human chromosome 3 and rodent chromosomes. The entire panel has 9 short arm breakpoints that involve bands p24.2, p22, p21, p14, and p12 plus a total of 11 long arm breakpoints that involve bands q13, q21, q25, q26, and q27. In addition, two cell lines appear to have breakpoints at or near the centromere. To date, we have used this panel to localize 92 sequences regionally on the short arm, 89 sequences on the long arm, and 7 sequences near the centromere. These hybrids are useful tools that allow the rapid localization of markers on chromosome 3 and greatly assist other mapping efforts.     

Benign neoplasms in the parotid gland in the county of Copenhagen 1986-1995

In a ten year period from 1986-95 433 patients were submitted to operation due to benign tumours in the parotid gland in Copenhagen County. In this study we made a retrospective analysis of the surgical outcome. The incidence was calculated to be 6.8 benign tumours in the parotid gland per 100,000 persons per year. The histological distribution showed a frequency of 54% pleomorphic adenomas, 28% adenolymphomas and 18% other tumours. In the observation period there was a 3% recurrence rate in our material, most frequently the pleomorphic adenoma. The risk of permanent damage to the facial nerve was 1% for severe injury and 3% for affection of the ramus marginalis. Frey's syndrome was present in 50% of the patients, moreover the syndrome was observed many years after surgery. The frequency rate reached its highest level about five years postoperatively.     

Effects of 6-hydroxydopamine and pimozide on temperature maintenance by the chicken

Intraventricular implants of pimozide in adult white leghorn hens were used to block dopamine (DA) receptors, and 6-hydroxydopamine (6-OHDA) was injected intraventricularly to destroy the noradrenergic system locally. The hens were exposed to ambient temperatures of 5 and 35 degrees C, and their core temperature was measured. One hundred micrograms of 6-OHDA significantly reduced the norepinephrine (NE) but not the DA content of the hypothalamus and reduced the uptake of [3H]NE but not of [3H]DA by synaptosomes in vitro. Neither of the drug treatments nor their combination affected average core body temperature (Tb) at either 5 or 35 degrees C. Pimozide treatment caused a lower maximum Tb at 35 degrees C and a higher maximum Tb at 5 degrees C than the control treatment. No evidence was obtained that 6-OHDA treatment affected body temperature regulation. It is concluded that neither the DA nor the NE system is essential for normal temperature maintenance in the hen exposed to either 5 or 35 degrees C.     

Whole body computed tomography in the diagnosis of abdominal and thoracic malignancy: review of 600 cases

Whole-body computed tomography (CT) was performed on 600 patients with known or suspected malignancy at the National Institutes of Health. This new diagnostic modality utilizes a finely columnated X-ray source mounted opposite a computer-linked detector bank to generate anatomic cross-sectional images of the body part being scanned. CT proved valuable in defining many pathologic processes and was, at times, deemed the only test short of exploratory surgery that yielded diagnostic information. This latter benefit was most acutely emphasized in lesions affecting the retroperitoneum, adrenal glands, pelvis, liver, and immediate subpleural pulmonary parenchyma.     

Analysis of empyrical factors for the determination of cardiac output

With a method by which no recirculation was possible the area of a dilution curve was obtained. On this basis it was searched what percentage represents the forward triangle of the total surface of the curve. The r?le of the V/F of the sampling system on this portion of the curve was studied, surface from which the empirical constants for the estimation of the cardiac output derivates. The data obtained suggest that the forward surface triangle was V/F dependent V/F less than 1 gave smaller forward triangles, the opposite was true with V/F greater than 1. It was also found that V/F greater than 1 subestimate the total surface of the dye curve, which means cardiac outputs higher than the real. The forward surface of the curve was reduced to a minimal percentage of the total curve when a V/F less than 1 was used. Situation that makes this area non useful to obtain from them empirical factors. If an empirical factor is use in order to measure cardiac output it had to be accepted that an important grade of error is introduced.     

The effects of roasting temperature applied to whole soybeans on site of digestion by steers: II. Protein and amino acid digestion

A 4 x 5 Youden square design was used to determine the effect of roasting temperature of whole soybeans on escape of CP from the rumen and disappearance of N from the small intestine in steers. Four steers (average BW 373 kg +/- 30.7) cannulated at the rumen, duodenum, and ileum were fed each of five diets (1.8% of BW) over five periods. The basal diet contained corn silage (50% of diet DM), alfalfa hay (24%), corn-urea premix (6%), corn starch grits (16.6%), and soybean oil (3.4%). Soybeans (16% of diet DM), either raw or heated to an exit temperature of 141, 149, or 157 degrees C in a commercial roaster, replaced the soybean oil and most of the corn starch grits in the soybean-containing diets. Ruminal ammonia N was lower (P < .05) for the basal diet than for the soybean-containing diets. Roasting temperature of whole soybeans had no significant impact on ruminal ammonia N. Total N reaching the duodenum was greater (P < .05) for steers fed the soybean diets than for steers fed the basal diet. Non-bacterial N (dietary N) at the duodenum was increased (P < .05) by feeding soybeans. Soybean N reaching the duodenum decreased with increased roasting temperature. Increased roasting temperature of whole soybeans seemed to make the soybeans more brittle, subsequently increasing degradation of CP in the rumen. However, disappearance of soybean N in the small intestine, as a proportion of the soybean N entering the small intestine, increased with increased roasting temperature. Apparent total tract N digestibility was increased (P < .05) by feeding soybeans. Flow to the duodenum and small intestinal digestibility of total, essential, and nonessential amino acids increased (P < .05) when soybeans were fed and when roasted vs raw soybeans were fed. Under normal roasting conditions, it seemed that little potential for heat damage to the soybean protein existed. Instead, undesirable effects of heating on handling characteristics of the soybeans were reached before the point at which loss of nutritive value occurred. Feeding steers diets containing roasted whole soybeans increased the N and amino acids available to steers over steers fed diets containing raw soybeans due to increased ruminal soybean nitrogen escape and increased small intestinal digestibility.     

Effect of polyamines and cystic fibrosis serum on glucose transport

The effect of plasma or serum from homozygotes and heterozygotes for the cystic fibrosis (CF) gene on the active uptake of 3-0-14C-methyl-D-glucose (3-0-14C-MDG) by rat jejunal epithelium was studied. Furthermore, the role of the polyamine, spermidine, and its products of metabolic degradation on glucose transport were investigated, and a relationship to the pathogenesis of membrane dysfunction in cystic fibrosis was postulated. Glucose transport in everted rat jejunal rings was used in the study. Results were expressed as 3-0-14C-MDG concentration ratio between the intracellular (ICF) and the extracellular fluid spaces (ECF) of the jejunal rings at the end of a 60 min incubation period. The mean ratio obtained from incubations of the rat jejunal rings in medium consisting of Krebs-Ringer-bicarbonate buffer and the labeled sugar was considered as 100% uptake. When plasma or serum, with or without spermidine, was mixed with the medium in a volume ratio of 1:3, a decrease in the active uptake of 3-0-14C-MDG was observed, expressed as percent inhibition. Percent inhibition of 3-0-14C-MDG uptake obtained when the rat jejunal rings were incubated in normal plasma was compared to that obtained with plasma from cystic fibrosis genotypes. It was found that: 1) plasma from 25 homozygous children had greater inhibitory effect on glucose uptake than plasma from 26 normal children; 2) plasma from 9 heterozygous women had greater inhibitory effect than that from 6 normal women; 3) the inhibitory effect of plasma from 3 homozygous children was not influenced by dialysis; 4) the inhibitory effects of paired plasma and serum samples from 9 homozygotes were comparable; 5) spermidine added to the incubating electrolyte solution did not affect glucose transport; 6) the addition of spermidine to reaction mixtures containing normal plasma potentiated the inhibitory effect; and 7) mixing and incubation of fresh bovine serum with reaction mixtures containing plasma from homozygotes decreased the inhibitory effect. The predominant inhibitory effect of plasma or serum from homozygotes and heterozygotes for the CF gene appears to be related to a nondialyzeable molecule(s). It does not seem to reflect the presence of high plasma glucose levels in cystic fibrosis nor to be the result of competitive inhibition between sugars. It does not seem to be the result of sodium or other electrolyte differences. A similar inhibitory effect is acquired by normal plasma after the addition of spermidine. On the other hand, plasma from CF homozygotes loses its inhibitory effect after incubation with fresh bovine serum. These findings may indicate that products of metabolic degradation of spermidine are responsible for the inhibitory effect of glucose transport and suggest the possibility of a role in abnormal polyamine metabolism in the pathogenesis of cystic fibrosis.