Hprt mutant frequency and GSTM1 genotype in non-smoking healthy individuals.

The T-cell cloning assay which combines mitogen- and growth factor-dependent expansion of lymphocyte clones with thioguanine selection of hypoxanthine-guanine phosphoribosyl transferase (hprt)-negative cells has been extensively used for studying human somatic gene mutation in vivo. However, large interindividual variations in the hprt mutant frequency (MF), much of which is not explained by donor attributes such as age and smoking habit, and interlaboratory variations in the experimental methodology, including cloning efficiency (CE), call for further developments of the cloning protocol and additional population studies. Using an improved T-cell cloning method, we have studied in vivo hprt MF of 76 non-smoking healthy males aged 23-77 years.

Relation between local acoustic parameters and protein distribution in human and porcine eye lenses.

The purpose of this study is to characterize the eye lens (human, porcine) by acoustic measurements and to investigate whether relations exist with the local protein content. The acoustic measurements were performed with a 'scanning acoustic microscope' (SAM), operating at a frequency of 20 MHz. At this frequency the lateral resolution in the acoustic images was 150 microns.

A new method to correlate acoustic spectroscopic microscopy (30 MHz) and light microscopy.

A powerful new method is used to investigate the correlation between light microscopic and acoustic properties of biological tissues. Specimens of liver were sectioned into successive slices, 250 micrometers and 10 micrometers thick. The thick sections were investigated acoustically, the thin sections by means of light microscopy.

Correlation of histology and acoustic parameters of liver tissue on a microscopic scale.

The correlation of several acoustic parameters with histological features was investigated in healthy White New Zealander rabbit liver (n = 10). Thin sections (10 microns) were studied by means of a light microscope in combination with a digital image processing system. Adjacent thick sections (250 microns) were studied by means of a custom-designed acoustic microscope.

Acoustic velocity and attenuation of eye tissues at 20 MHz.

The ultrasound velocity and frequency-dependent attenuation of human and porcine eye tissues (cornea, lens, retina, choroid, sclera, vitreous body) were measured in the frequency range from 17 to 23 MHz. The results for the ultrasound velocity were compared to values taken from the literature and appeared to be in the same range. A comparison made between the acoustic parameters of human and porcine eyes showed that the porcine eye can serve as an animal model for the human eye.

Ultrasonic spectroscopy of the porcine eye lens.

The purpose of the work is to measure and study the acoustic characteristics of the porcine eye lens and find correlations with chemical and optical parameters, obtained from literature. Ultrasonic spectroscopy was performed by using a scanning acoustic macroscope (frequency 20 MHz, resolution 150 microns). The transducer performed a two-dimensional scan over a central slice (1 mm thickness) of porcine lens (number of lenses = 10).

Hprt activities and RNA phenotypes in 6-thioguanine resistant human T-lymphocytes.

The phenotypic effects of mutation in the hypoxanthine phosphoribosyltransferase (hprt) gene on hprt enzyme activity and hprt mRNA levels were studied in 6-thioguanine (TG) resistant human T-cell clones with various types of hprt mutation. The mean enzyme activity in 16 TG selected clones was less than 1% of the mean in unselected clones. The hprt mRNA levels, measured by a quantitative RNA/RNA solution hybridization assay, were within the normal range in 38% of the mutant clones.

Molecular spectrum of background mutation at the hprt locus in human T-lymphocytes.

The molecular basis of somatic mutation at the hypoxanthine-guanine phosphoribosyl-transferase (hprt) locus in human 6-thioguanine resistant T-cell clones from 17 individuals has been studied by Southern blot analysis, multiplex PCR (polymerase chain reaction) and direct sequencing of PCR amplified hprt cDNAs or genomic DNA. Twenty-three novel mutations were detected, which in addition to previously described mutations provide a background mutational spectrum based on a total of 45 hprt mutations in human T-cells. Twenty T-cell mutants had base substitutions in the coding region leading to 15 missense and five nonsense mutations.

Missense mutations and evolutionary conserved amino acids at the human hypoxanthine phosphoribosyl-transferase locus.

Molecular characterization of in vivo mutation at the human hypoxanthine phosphoribosyltransferase (hprt) locus has revealed a broad spectrum of mutation, both with regard to germ-line mutation in Lesch-Nyhan and gout patients, and somatic mutation in 6-thioguanine resistant T-lymphocytes from healthy individuals. The pattern of missense mutation shows a non-random distribution with a preferential location to codons for amino acids which are identical in human and the two parasites Schistosoma mansoni and Plasmodium falciparum. Although these 'evolutionary conserved' amino acids account for only 32% of the amino acids in the human hprt protein, they are involved in 76% of the missense mutations at the hprt locus in human T-lymphocytes, 67% in Lesch-Nyhan patients (with severe hprt-deficiency), but only 43% in gout patients (with partial hprt deficiency).

Mutation analysis and prenatal diagnosis in a Lesch-Nyhan family showing non-random X-inactivation interfering with carrier detection tests.

A nonsense mutation at the CpG-site in the codon for Arg(169) in the gene for hypoxanthine phosphoribosyltransferase (hprt) was identified by genomic polymerase chain reaction (PCR) and DNA sequencing in cultured fibroblasts from two brothers with Lesch Nyhan's syndrome. The recurrence of mutation at this CpG-site in several unrelated Lesch-Nyhan families suggests that deamination of 5-methylcytosine is a possible mechanism for mutagenesis. The level of hprt-mRNA in the fibroblasts of the patients was similar to that in healthy controls, whereas hprt-enzyme activity was not detectable.

In vitro classification of gallstones by quantitative echography.

Gallstones (n = 20) were classified by dual energy Computer Tomography (CT) into three main classes: pure cholesterol stones (I), combination stones (II) and calcium stones (III). Further subclassification was possible by using morphological criteria. The acoustic measurements that were performed were measurements of the velocity of sound, the attenuation coefficient slope with frequency and intercept at 4.

Effects of tissue processing techniques in acoustical (1.2 GHz) and light microscopy.

In this study the influence of various tissue processing and staining techniques on the acoustical properties of liver tissue was investigated. A qualitative study was performed using ultrasound attenuation as the imaged parameter of a combined optical/acoustical microscope with a 1.2 GHz transducer.

The fragile X mutation does not have any major effect on the expression of the hypoxanthine phosphoribosyltransferase (HPRT) locus in human fibroblasts.

The possible influence of the fragile X mutation at Xq27 on the expression of the neighbouring gene (at Xq26) for hypoxanthine phosphoribosyl transferase (HPRT) was studied by determination of the levels of HPRT-RNA and HPRT enzyme activity in fibroblast cell cultures from 7 fragile X patients. These levels were lower (although not statistically significantly lower) than in normal fibroblast cultures. Hence, these data do not support the notion of a major effect of the fragile X mutation on the expression of the HPRT gene.

Place of delivery in The Netherlands: actual location of confinement.

Preferred and actual locations of confinement were compared in a group of 170 nulliparous women. Voluntary changes in preferred location for birth were rare and concerned only changes from hospital to home confinement. Obligatory changes due to referral to consultant obstetricians occurred frequently: 58.

Expression of the hypoxanthine phosphoribosyl transferase gene in resting and growth-stimulated human lymphocytes.

Expression of the hypoxanthine phosphoribosyl transferase (hprt) gene was studied in resting and growth stimulated human lymphocytes by determinations of hprt-RNA and enzyme activity levels in cell extracts. Hprt-RNA was determined by quantitative solution hybridization and enzyme activity by measuring the rate of conversion of [14C]hypoxanthine to inosine 5'-monophosphate. In resting Go-lymphocytes, the hprt activity (5 nmol/h per 10(6) cells) was twice as high as in human fibroblasts, whereas the hprt-RNA level was very low (0.

Influence of attenuation on measurements of ultrasonic myocardial integrated backscatter during cardiac cycle (an in vitro study).

The purpose of this study was to investigate the dependence of ultrasonic integrated backscatter (IB) and attenuation in myocardium on wall thickness in a state of acute ischemia. Therefore, an in vitro experiment was set up in which attenuation, IB and wall thickness of a piece of freshly excised myocardium could be measured almost simultaneously. The myocardium was taken from 11 Yorkshire pigs (25-30 kg) that were killed less than 45 min before the experiment.

Esophageal cancer.

The authors invited eminent Belgian specialists to report on their experience in esophageal cancer. A summary is given of the most striking features of their reports.

Conventional X-ray examination in esophageal cancer: an opinion.

Double contrast is the best radiological technique for the detection of small esophageal cancer. Gastrografin is indicated when a blind mediastinal fistula is suspected; in cases of choking or suspicion of a fistula with the airways, a low osmotic hydrosoluble Iodium compound must be used. Conventional radiology and endoscopy are complementary techniques for the detection of esophageal cancer, as some lesions may be missed or misinterpreted by both.

Influence of histochemical preparation on acoustic parameters of liver tissue: a 5-MHz study.

In this study the influence of various histological techniques on the acoustic parameters of liver tissue was investigated. Radiofrequency (RF) echographic data were obtained in vitro from 21 liver samples taken from 8 white New Zealander rabbits. The samples were measured in four different subsequent histological tissue processing conditions (freshly excised, 4% buffered formalin fixed, after it went through a paraffin cycle and after staining with hematoxylin and eosin).