Evolution of genetic variability in a population of the edible snail, Helix aspersa Müller, undergoing domestication and short-term selection.

The evolution of genetic variability is studied in six successive generations of a population originating from wild Helix aspersa. During the first three generations (G1 to G3), no artificial selection was applied. During the next three generations (G4 to G6), two lines were reared: a control line (C) and a line (S) selected for increased adult weight. Genetic variability is described by genealogical parameters (inbreeding, number of founders, effective number of founders and ancestors, effective number of remaining genomes) and by the additive genetic variance in adult weight. A large decrease in all parameters was observed between G1 and G2, suggesting strong natural selection: additive genetic variance in adult weight (transformed data) decreased from 0.0119 +/- 3.8 x 10(-3) to 0.0070 +/- 1.7 x 10(-3) (P < 0.05) and effective number of ancestors from 97.4 to 67.0. Selection also caused a large decrease during the first generation: additive genetic variance was 0.0079 +/- 2.1 x 10(-3) in G3 and 0.0040 +/- 1.1 x 10(-3) after the first selection cycle (P < 0.02). At the same time, the effective number of ancestors decreased from 59.2 to 29.5 and 24.2. This decrease is consistent with the theory of selection and the Bulmer effect.

Comparison of different electrophoretic separations of hen egg white proteins.

The hen egg white protein composition has not yet been fully defined. To improve the knowledge of this biological fluid, the most usual and recently developed electrophoretic methods have been used: SDS-PAGE, native-PAGE, isoelectric focusing (IEF), and 2-dimensional electrophoresis (2DE). Seven of the major known proteins were thus identified in at least one electrophoretic system. Isoforms of ovotransferrin, ovalbumin, and ovomucoid were visualized when pI was used for the separation. Two-dimensional electrophoresis allowed separation of a very large number of spots. In each of the four systems, some components were revealed but not identified, and unknown spots were particularly numerous with 2DE. With this technique, many spots corresponding to small acidic proteins were highlighted, among which was the Ch21 protein, whose presence in hen egg white was thus confirmed. This study thus constitutes, to our knowledge, the first proteomic investigation of hen egg white.

Stearoyl-CoA desaturase 1 coding sequences and antisense RNA affect lipid secretion in transfected chicken LMH hepatoma cells.

Hepatic stearoyl CoA desaturase (SCD) activity in chickens from a fat line is higher than that of chickens from a lean line and correlates with plasma triacylglycerol concentrations. Furthermore, in these lines, the hepatic SCD1 mRNA level is positively correlated with the adipose tissue weight. To analyze the contribution of the SCD1 gene in the regulation of adiposity in the early stages of triacylglycerol secretion, SCD1 coding sequence and antisense RNA expression vectors were transfected in LMH cells. After selection, these cells were analyzed with regard to SCD1 expression and lipid secretion. The amounts of secreted triacylglycerols and phospholipids were shown to be higher in LMH cells transfected with the SCD1 gene, but reduced in those transfected with the SCD1 antisense sequences when compared to cells transfected with the vector alone (without SCD1 sequences). These results provide direct evidence that the expression of the SCD1 gene plays a major role in the triacylglycerol and phospholipid secretion process.

Direct and correlated responses to individual selection for large adult weight in the edible snail Helix aspersa Muller.

A selection experiment for large adult weight based on individual performance was conducted for three generations in Helix aspersa aspersa. A second line was kept as an unselected control line. Direct response measured as deviation from the control line was 3.55 g after three generations of artificial selection, which averaged 13%. Realized heritability was 0.38 +/- 0.04. Correlated responses to selection showed a significant increase in weight after hibernation, mean egg weight and mean weight of newly-hatched snails with selection. For adult age, egg number, and hatching rate, no significant change correlated to selection was found, but this is to be confirmed.

Characterization of the chicken fatty acid synthase gene 5' part and promoter region.

Fatty acid synthase activity has been shown to be regulated mainly at the transcriptional level under both dietary and hormonal influences. As a first step towards elucidating the factors involved, we isolated and characterized chicken genomic clones encompassing the 5' part of the chicken fatty acid synthase gene and its flanking region. The entire region of the cloned DNA spans 30 kb, and the first three exons of the gene were mapped to a 6.3-kb genomic fragment. The transcription initiation site was determined after subcloning the cDNA which encodes the 5' end of the mRNA. The first exon, which was 129 bp long, was located approximately 5.3 kb upstream of the second exon, which contained the start codon. In the 5' flanking region, putative TATA and CAAT boxes were located 30 and 92 bp, respectively, upstream of the transcription initiation site. The 5' flanking region contained numerous sequences corresponding to consensus binding sites for transcription factors. Various lengths of flanking sequences extending up to 1028 bp upstream of the transcription initiation site and containing 100 bp of the first exon were linked to the bacterial chloramphenicol acetyltransferase gene; in this study, these constructs were analyzed in transient transfection assays in human hepatoma cells. The proximal 125-bp sequence upstream of the transcription start site was shown to be a basal promoter. The cloning and characterization of the chicken fatty-acid synthase gene provides some further insight into the regulation of fatty acid synthesis in birds as compared to mammals.

Cloning and characterization of the 5' end and promoter region of the chicken acetyl-CoA carboxylase gene.

Acetyl-CoA carboxylase is a rate-limiting enzyme in the biogenesis of long-chain fatty acids. In the present study, the 5' end and flanking region of the acetyl-CoA carboxylase (ACC) gene was analysed in the chicken. A genomic clone was isolated containing the first three exons, the third one containing the ATG codon. Using nuclease-mapping experiments and primer-extension analyses, the transcription-initiation site was located 153 nucleotides upstream of the ATG codon. In contrast with rat ACC gene expression, reverse transcriptase PCR analysis performed on chicken liver mRNA did not reveal alternative splicing in the 5'-untranslated region of these messengers. The promoter region is very G+C rich, and contains no TATA or CAAT boxes. Analysis by transient transfection in a human hepatoma cell line (HepG2) demonstrates that the promoter activity requires the presence of symmetrical sequences located upstream of the GC boxes. Transcription of this gene is found to be controlled by tri-iodothyronine in HepG2 cells, but the sequence responsible for the tri-iodothyronine response is not the consensus tri-iodothyronine-responsive element localized in the promoter. These results bring new insights to the regulation of the chicken ACC gene which differs from that of the rat.

Lipogenic enzyme and apoprotein messenger RNAs in long-term primary culture of chicken hepatocytes.

Hepatocytes isolated from 9-week-old chickens were cultured in a serum-free, hormonally defined medium. Relative amounts of mRNAs coding for lipogenic enzymes (acetyl-CoA carboxylase, fatty acid synthase, delta 9 desaturase, malic enzyme) and apoproteins (apoprotein A1 and apoprotein B) were determined until the 12th day. beta-actin and albumin mRNA, as well as albumin secretion, were also assessed. Cellular metabolic activity appeared to be very low for the first days of culture, but increased after the 7th day. All the mRNAs studied, except for that of malic enzyme, were present from this time throughout the culture lifespan. The biological significance of the observed results and the relevance of this chicken hepatocyte culture system for long-term metabolic and genetic studies are discussed.

Study of tissue compression in breast phantoms: possible implications for the use of X-ray mammography as a method of imaging breast carcinoma.

A method is described of constructing a simple phantom breast containing an idealised tumour with elastic properties. The transparent tissues are compressed and evidence is adduced to show that with tissue compression the tumour boundary is subjected to a degree of strain the magnitude of which is dependent on the degree of deformation. Excessive compression can lead to fracture of the tumour. A literature survey reveals that malignant tissues are more delicate than normal tissues. The study underlines the need for care when using X-ray mammography with compression to image a cancer bearing breast, in order to avoid disseminating cancer cells via the circulation.

Identifying genes involved in the variability of genetic fatness in the growing chicken.

A precise knowledge of the genome involved in the expression of a quantitative trait could provide a useful tool in breeding programs; molecular genetic methods are capable of yielding this kind of information. An experimental procedure is presented here for identifying genes whose expression is related to weight variability of abdominal adipose tissue in the growing chicken. Quantitative traits are the result of metabolic pathways exhibiting some major regulation stages that are controlled genetically. These steps involve genes that may act as "major genes". With regard to chicken fat metabolism, most fatty acids are synthesized in the liver and incorporated into very low density lipoprotein (VLDL) particles before their secretion into the plasma. Accordingly, the present study focused on the expression of liver genes. The mRNA of lipogenic enzymes (acetyl-coenzyme-A carboxylase, fatty acid synthase, malic enzyme, and delta 9-desaturase) were analyzed. Also studied were apoprotein (apo)A1, apoVLDL-II, and apoB mRNA from 9-wk-old male chickens from two lines selected for high and low abdominal fat pads. Significant differences for apoA1 mRNA levels occurred between fat and lean birds. Moreover, the total quantity of mRNA provided an accurate estimation of the abdominal fat pad (r = .74 with P < .05).

Measurement of blood flow in tibial fracture patients using positron emission tomography.

The quantification of local bone blood flow in man has not previously been possible, despite its importance in the study of normal and pathological bone. We report the use of positron emission tomography, using 15O-labelled water, to measure bone blood flow in patients with closed unilateral fractures of the tibia. We compared fractured and unfractured limbs; alterations in blood flow paralleled those found in animal models. There was increased tibial blood flow at the fracture site as early as 24 hours after fracture, reaching up to 14 times that in the normal limb at two weeks. Blood flow increase was less in displaced than in undisplaced fractures. The muscle to bone ratios of blood flow were similar to those in previous animal work using other techniques. Positron emission tomography will allow study of human bone blood flow in vivo in a wide variety of pathological conditions.

Isolation and long-term maintenance of differentiated adult chicken hepatocytes in primary culture.

Adult chicken hepatocytes were obtained by an adaptation of the two step in situ collagenase perfusion. Usually 0.5 to 1 x 10(9) cells were obtained, with 75 to 95% viability. Hepatocytes attached within 2 h when plated on plastic cell culture dishes and spread in 4 h, surviving for several months in a specific serum-free medium. These cells retained a typical parenchymal cell morphology and the ability to produce a specific protein (albumin) throughout the culture period. We hereby provide a suitable model for studying hepatic metabolism in birds.

Pattern of 2-deoxy-2-[18F]-fluro-D-glucose accumulation in liver tumours: primary, metastatic and after chemotherapy.

Eight patients with liver metastases from adenocarcinoma of the colon or rectum, two with suspected hepatic metastases and one with primary hepatoma were studied with 2-deoxy-2-[18F]-fluro-D-glucose (18F-FDG) using positron emission tomography (PET). In five of the patients with metastatic tumour a second examination was performed four weeks after treatment with recombinant interleukin-2 (rIL2) and fluorouracil (5FU). In all tumours (one primary and eight metastatic) the radioactivity was seen to accumulate in a rim around each tumour with a large central area showing no uptake. In the five cases imaged after treatment with rIL2, the appearance of the tumour uptake was the same as before treatment. In the two cases of suspected but not proven metastases, no abnormal accumulation of 18F-FDG was seen.

Theoretical considerations in relation to the treatment of brain tumours by means of local hyperthermia generated by ultrasound fields.

Brain tumours comprise a significant fraction of all tumours in the human body. Despite the development of technology in clinical oncology, these tumours still present a difficult challenge. The margin between destruction of tumour and damage to normal tissue is narrow in the brain. The price paid for producing tissue damage outside the tumour is high in terms of quality of survival. Results of many experiments with ultrasound hyperthermia show that this new technique is successful for treating certain types of malignant tumours. In the case of brain tumours, applying focused fields should have the advantage of selectively destroying the tumour and leaving surrounding tissues intact. Previous attempts at applying ultrasound to the field of neurosurgery are reviewed. Those factors which have effects on the generation of thermal fields in brain tissues are considered in relation to the possible treatment of human brain tumours. Calculated thermal fields in a 2-D brain model generated by an applicator built in our laboratory are also presented.

AURA: a new respiratory monitor and apnoea alarm for spontaneously breathing patients.

Previous attempts to introduce non-invasive monitoring of ventilation of spontaneously breathing patients into routine practice have been unsuccessful. The Aberdeen University Respiratory Alarm (AURA) allows such monitoring by utilizing the pyroelectric property of polarized polyvinylidine fluoride sensors to detect temperature changes that occur during breathing into an oxygen delivery face mask. A quartz crystal oscillator generates pulses that allow measurement of interexpiratory time and ventilatory frequency. The system incorporates LED digital displays, a bargraph and audiovisual alarms. An analog output permits display and analysis of the sensed signals. AURA performed satisfactorily in both volunteer studies and six patients in the clinical setting. AURA may be an appropriate respiratory transducer in those patients requiring oxygen therapy.

Some of the factors involved in the Sarvazyan method for recording ultrasound field distributions with special reference to the application of ultrasound in physiotherapy.

The mapping of ultrasound fields using the dye paper method is described. The mode of action by which the dye concentration is increased has been investigated. Microbubbles on the paper surface and microstreaming of dye solution seem to be essential features of the mapping method. The streaming is thought to inhibit the formation of a dye depletion layer near the paper to an extent dependent on local sound intensity A linear relation between streaming velocity and intensity is derived.

Impedance imaging in upper arm fractures.

We used the Aberdeen impedance imaging system to drive a constant current of 1 mA on a 10 kHz sine wave into the upper arm encircled by an elastic belt of 16 equi-spaced strip electrodes. The system was used to examine a normal upper arm, an upper arm with a recent humeral fracture and an upper arm with a clinically united fracture. We approximated the human upper arm to a circular cylinder and assumed bilateral symmetry of normal human limbs. We measured transverse limb resistivity ratios and reconstructed static two-dimensional images of the spatial distribution of log(resistivity) by the equipotential back projection technique using a homogeneous muscle equivalent saline reference. Our results indicate that impedance osteography provides unique information about the changing electrical characteristics at the fracture site. This information could prove a useful adjunct to clinical and radiological tests for fracture union.

AURA: a new respiratory monitor.

The Aberdeen University Respiratory Alarm (AURA) is designed to monitor the respiratory rate and the inter- expiratory period (respiratory gap) in the spontaneously breathing patient using polarised polyvinylidene fluoride sensors incorporated in the Oxygen delivery mask. The monitor is composed of three modules--the sensor module includes a pair of polyvinylidene film strips mounted on the inner surface of the Oxygen mask. The difference in temperature between the ambient Oxygen enriched air and the warmer expired air allows the pyroelectric property of the sensors to be harnessed. The sensor arrangement allows a differential signal to develop between them which is amplified, filtered and rectified prior to triggering a counter. The timing module is based on a 32,768 Hz crystal oscillator that generates pulses allowing the period between expirations to be measured and compared to the maximum allowable period set by the user. The respiratory rate is also measured and compared to the minimum set by the user using EPROMS and comparators. The display module consists of a bar-graph display of the level of the sensed signal with each breath with a soft audible click. Eight segment LEDs are used to provide a digital display of the respiratory rate and the respiratory gap. Audiovisual alarms are triggered when the rate falls short of the set rate and/or when the gap exceeds the set gap. The need for continuous respiratory monitoring of the spontaneously breathing adult in hospital practice led us to develop this prototype with sensors whose design and configuration can be considerably improved.

ApoVLDLII gene transcription in immature cockerels without estradiol stimulation.

1. Previous studies have led to the conclusion that the gene encoding the apo very low density lipoprotein II (ApoVLDLII) is under full estrogen control. However, by using a sensitive hybridization technique we found a weak expression of this gene in some males in the absence of hormone. 2. As the vitellogenin II gene, which is also estrogen dependent, remains inactive in these animals it is likely that they carry a deregulated allele of the ApoVLDLII gene. 3. The highest level of ApoVLDLII mRNA is found in genetically fat males suggesting a possible involvement of the ApoVLDLII gene in fatness determinism.

Quantitative imaging in vivo. Why? What is being done now? What are some of the problems? What may the future be?

The visual system is highly sophisticated, yet at the task of quantifying the information in images it is very easily misled. It is probably for this reason that medical imaging has often concentrated on the visual analysis of images and played down the value of extracting quantitative information. Yet nearly all modern medical imaging techniques use digital technology to some degree and numerical information can be readily extracted. Unless more use is made of this quantitative data, medical imaging will never make the transition from an art into a science.