Molecular events including p53 and k-ras alterations in the in vitro progression of a human colorectal adenoma cell line to an adenocarcinoma

The aim of the current study was to identify genetic abnormalities in human colorectal adenoma and carcinoma derived cell lines, and to determine whether the genetic changes which occur in vitro are relevant to the in vivo situation. Loss of 1p(33-35) region was shown to be the most common chromosome 1 abnormality and loss of heterozygosity (LOH) of the DCC gene and/or adjacent sequences was detected in all adenoma derived cells as well as the carcinoma cell lines. The level of p53 protein was also investigated as increased cellular p53 protein had previously been associated with mutation of the p53 gene. A further aim was to investigate genetic changes in our in vitro model of tumour progression, where the adenoma derived PC/AA cell line has previously been converted in vitro to two distinct tumorigenic phenotypes, producing either an adenocarcinoma or a mucinous carcinoma in athymic nude mice. Progression to the adenocarcinoma phenotype was shown to involve a specific chromosome 1 rearrangement, loss of both normal copies of chromosome 18 (although DCC gene sequences were retained), loss of the remaining wild type allele of k-ras resulting in homozygosity for the k-ras codon 12 mutation and increased cellular p53 protein as detected by SDS-PAGE Western blotting. The increase in p53 protein was shown not to be due to the acquisition of a mutation in the p53 gene. Interestingly, progression of the adenoma derived PC/AA cell line to the mucinous malignant phenotype did not involve any of these molecular rearrangements, suggesting that different genetically distinct pathways are involved in colorectal carcinogenesis. These studies show that the genetic changes in our in vitro model of human colorectal tumour progression are similar to those observed in in vivo studies.     

Effect of charcoal-broiled beef on phenacetin metabolism in man

When charcoal-broiled beef was fed to human volunteers, who were then given phenacetin orally, the concentration of phenacetin in the plasma was lowered, but its half-life in the plasma was not changed. The data suggest that feeding charcoal-broiled beef enhances the metabolism of orally administered phenacetin in the intestine or during its first pass through the liver, or both.     

Serum albumin. A CAP survey

The results of a 1974 survey of albumin measurements as performed by more than 1,300 laboratories are presented. The most widely used methods are the dye-binding technics: bromcresol green (BCG) and 2-(4'-hydroxyazobenzene) benzoic acid (HABA). These are followed by electrophoresis and salt fractionation. All methods yielded comparable albumin concentrations except electrophoresis, which manifested a consistent low bias. This close agreement is attributed, in part, to the normal-range concentration of albumin in the test specimen. Type of standardization, i.e., commercial serum, bovine serum albumin, human serum albumin, or pooled serum, did not appear to be a factor in the estimation of albumin in the normal serum submitted for analysis. Surprisingly, interlaboratory variation, from method means, was the lowest for salt fractionation and electrophoretic technics.     

Calculation of coronary blood flow from myocardial clearance of systemically administered 133Xe

Clearance curves for arterial and coronary-venous blood were determined after systemic left-ventricle or pulmonary-artery infections of 133Xe, paired with selective left-coronary-artery injections of 133Xe in 20 dogs with closed chest. Coronary blood flows calculated from systemic and coronary-artery injections were comparable only when a correction was made for arteriol recirculation of 133Xe following the systemic injection (r = 0.962) for left ventricle and 0.932 for pulmonary artery, paired with coronary artery). Experiments in four other dogs, verified that clearance of 133Xe from the pulmonary circulation was only about 60%. The myocardium/blood 133Xe partition coefficient, determined in vivo in ten dogs, agreed within 10% with that previously determined in vitro.     

Studies on the antitesticular action of DL-6-(N-2-pipecolinomethyl)-5-hydroxy-indane (PMHI) in the rat

Both prepubertal and adult rats were treated with a single oral dose of either 60 mg or 120 mg of dl-6-(N-pipecolinomethyl)-5-hydroxy indane maleate (PMHI) per kg of body weight. Their testicular weights were drastically reduced compared with those of the controls. A follow-up, beginning on the third day post-treatment and continuing for a period of 50 days, showed that the body weight growth of PMHI-treated rats was not retarded. The hormonal profile indicated that, except for FSH which showed a transitory elevation in PMHI-treated immature rats, the serum levels of LH, estrogen, and testosterone were indistinguishable from those of the controls. Testicular histology revealed that the spermatogenic process in PMHI-treated rats recovered at a dose-related rate. EM sections of testes of adult rats indicated that cytoplasmic vacuolation appeared in the Sertoli cells 5 h post-treatment. The consequent cascade of arrested spermiogenesis included abnormal acrosomal condensation of spermatids and sloughing of polynucleated spermatids. Some spermatocytes also seemed to be affected, but spermatogonia and Leydig cells remained intact. These results indicate the PMHI acts primarily on Sertoli cells and causes arrest in the spermiogenetic stage of the spermatids. At a higher and toxic dose of PMHI, however, the earlier germinal elements might also be affected, due to the extensive damage to the supporting Sertoli cells.     

Clinical and biochemical studies in an American child with sialuria

Sialuria is a rare inborn error of sialic acid (NeuAc) metabolism resulting from failure of CMP-NeuAc to adequately feedback inhibit the rate-limiting enzyme in sialic acid synthesis, UDP N-acetylglucosamine (UDP-GlcNAc) 2-epimerase. We describe the fourth reported sialuria patient, T.W., whose clinical features include developmental delay, coarse facies, and massive urinary excretion of sialic acid. Biochemical studies of T.W. fibroblasts revealed a 200-fold increase in free NeuAc content compared with normal. Bound NeuAc was only slightly elevated. The free NeuAc was predominantly in the cytosol fraction of fibroblasts after differential centrifugation, with only 4% of the free NeuAc content in other (nuclear, granular, and microsomal) cellular compartments. CMP-NeuAc inhibited UDP-GlcNAc 2-epimerase by 80% in normal fibroblasts but inhibited the epimerase of T.W. (sialuria) cells by only 13%. Cytidine feeding of sialuria fibroblasts decreased the intracellular free NeuAc content by 47%; this was accompanied by a fourfold increase in CMP-NeuAc, which may be sufficient to feedback inhibit the mutant epimerase and reduce free NeuAc production. Cytoplasmic pH was determined by the pH sensitive fluorescent indicator 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein, pentaacetoxymethylester (BCECF/AM) using the H+ equilibration method. The intracellular pH of sialuria fibroblasts, 7.18 +/- 0.04, was not found to be significantly different from that of normal cells (7.19 +/- 0.08).     

Gamow-Teller matrix elements for two-neutrino double beta decay within a second quasi-random-phase approximation

The ascendency of the cerebral cortex in mammals naturally raises questions about the role of the archetypal subcortical centers we share in common with other phyla. Here we report a situation in which an ancient oculomotor control center, the nucleus of the optic tract, is not so much dominated by the cerebral cortex as served by it. We suggest that the organization of cortical output to subcortical centers may be helpful in understanding the function of the cerebral cortex.