Chromosome-specific microsatellite sets for fluorescence-based, semi-automated genome mapping

To facilitate large-scale genetic mapping of the human genome, we have developed chromosome-specific sets of microsatellite marker loci suitable for use with a fluorescence-based automated DNA fragment analyser. We present 254 dinucleotide repeat marker loci (80% from the Généthon genetic linkage map) arranged into 39 sets, covering all 22 autosomes and the X chromosome. The average distance between adjacent markers is 13 centiMorgans, and less than 4% of the genome lies more than 20 cM from the nearest marker. Each set of microsatellites consists of up to nine marker loci, with allele size ranges that do not overlap. We selected marker loci on the basis of their reliability in the polymerase chain reaction, polymorphism content, map position and the accuracy with which alleles can be scored automatically by the Genotyper program.     

Elemental distributions in predentine associated with dentine mineralization in rat incisor

Electron probe microanalysis was applied to study quantitatively and semi quantitatively the elemental concentrations and distributions that occur in predentine during the dentine mineralization of rat incisor. Apex regions of the continuously growing incisors were rapidly dissected and cryofixed in liquid nitrogen-cooled propane. Ultrathin cryosections were prepared from the dentine tissue. On the average in the extracellular predentine element concentrations of calcium and phosphorus were about 0.5% (w/w) and 0.5-1% (w/w), respectively; so the calcium content in the extracellular predentine is higher while the phosphorus content is much lower than in the odontoblast area. Due to the high content of glycosaminoglycans in the extracellular matrix the concentration of sulfur in the predentine was more than 1% (w/w); the potassium content was found in the range of 0.6-0.8% (w/w) which is quite high for an extracellular area and the concentrations of sodium and chlorine were higher than 2% (w/w). Elemental mapping analysis was carried out to demonstrate the distribution of some important elements at the predentine/dentine border during mineralization.     

The use of 99Tcm-Technegas in the investigation of patients with pulmonary thromboembolism

Pulmonary embolism remains a major cause of morbidity and mortality, being responsible for an estimated 200,000 deaths per annum in the USA and 21,000 per annum in the UK. Lung scintigraphy is in many instances the investigation of choice in suspected pulmonary thromboembolism. A normal perfusion lung scan excludes pulmonary embolism. An abnormal perfusion scan, while being sensitive, is of low specificity for the diagnosis of pulmonary embolism and needs to be complemented with a ventilation study. Lung ventilation has been studied using inert gases of radiolabelled aerosols. The new radiopharmaceutical 99Tcm-Technegas is a suspension of 99Tcm-labelled, ultrafine, carbon particles produced in an atmosphere of high-purity argon. The size of the particles is of the order of 0.005-0.2 microns, which assures good peripheral penetration and alveolar deposition. After inhalation, static images in multiple projections may be acquired. The resultant images are of excellent technical quality and several publications have shown the value of Technegas images as an adjunct to perfusion imaging in suspected pulmonary thromboembolism. Studies comparing Technegas images with other ventilatory radiopharmaceuticals have in the majority of instances reported comparable diagnostic qualities.     

Importance of intercostal artery reattachment during thoracoabdominal aortic aneurysm repair

PURPOSE: We studied the relationship of neurologic deficit to ligation, reimplantation, and preexisting occlusion of intercostal arteries to determine which arteries and consequent management are most critical to outcome in thoracoabdominal aortic aneurysm repair. METHODS: From February 1991 to July 1996, 343 patients with thoracoabdominal aortic aneurysms underwent repair by one surgeon. In this study, only Crawford types I, II, and III (n = 264) were considered. Of these, 110 (42%) were type I, 116 (44%) type II, and 38 (14%) type III. The adjuncts of distal aortic perfusion and cerebrospinal fluid drainage were used in 164 patients (62%). Data were analyzed by contingency table and by multiple logistic regression. RESULTS: Early neurologic deficit occurred in 23 patients (8.7%), and late deficit in 10 patients (3.8%). Neurologic deficit in patients with at least one reimplantation and no ligation of arteries T11 or T12 occurred in 19 of 147 (12.9%). Neurologic deficit for occlusion of the same arteries occurred in 11 of 111 (9.9%), whereas for ligation of T11 and T12 neurologic deficit occurred in three of six (50%; reimplantation, p < 0.03; occlusion, p < 0.006). In addition, reimplantation of intercostal arteries T9 or T10 was significantly associated with reduced late neurologic deficit in multivariate analysis (p = 0.05). No other intercostal artery status was associated with modification of the neurologic deficit rate. Multivariate analysis showed type II aneurysms and acute dissections to be significantly associated with an increased risk of postoperative neurologic deficit (p < 0.0009, 0.002, respectively). Adjuncts were protective (p < 0.007), most often in types II and III (14.1% neurologic deficit in type II with adjunct, 35.3% without; 0% in type III with adjunct, 20% without). CONCLUSION: Patients with patent arteries at the T11/T12 level have highly variable outcomes depending on whether the arteries are reattached or ligated. Our data suggest that reimplantation of thoracic intercostal arteries T11 and T12 is indicated when these arteries are patent. Reimplantation of T9 and T10 lowers the risk of late neurologic deficit, probably by decreasing the spinal cord's vulnerability to changes in blood and cerebrospinal fluid pressure in the days after surgery. Adjuncts lower overall risk and provide adequate time for targeted intercostal artery reimplantation.     

A new mouse model of experimental melanoma for vaccine and lymphokine therapy

The annual incidence of malignant melanoma is estimated at 10-12 per 100,000 inhabitants in countries of central Europe and the United States, and alarmingly there has been a dramatic upward trend in that estimate. The B16 murine melanoma is a rapidly growing metastatic tumor of spontaneous origin, as are human malignant melanomas. Melanoma cells produce specific antigens which are uniquely different from normal cellular antigens, and the expression of such antigens is the cornerstone for preparation of anti-melanoma vaccines. One major problem in evaluating the effectiveness of vaccination and other biologic therapies is the variability of experimental tumor models. A new metastatic model of experimental melanoma which was developed in our laboratory imitates the major clinical stages of malignant metastatic melanoma: stage I, primary (local) tumor growth and bone marrow invasion; stage II, regional lymph node involvement; and stage III, metastasis to distant organs, such as the lungs. This model has been used successfully for screening vaccines constructed in our laboratory. Immunization with formalinized vaccines (of extracellular antigens, intact melanoma cells, or B700 antigen) or irradiated vaccines (of intact melanoma cells) partially inhibit primary melanoma tumor growth, reduce metastasis to regional lymph nodes and lungs, and significantly increase mean survival time. These anti-tumor effects were improved when polyvalent and monovalent vaccines were combined with IL-2 therapy. We also compared the immunogenic activity of vaccines made from B16 melanoma cells transfected with genes encoding murine IL-2 or GM-CSF, and effects on tumor bearing mice were compared with or without therapy using the corresponding lymphokines. In sum, comparison of antibody production, growth of primary melanoma tumors, number of surviving mice, mean survival time, and percent of mice with lung metastases, showed that the best course of immunotherapy involves vaccination of mice with irradiated B16 melanoma cells transfected to secrete GM-CSF, coupled with GM-CSF therapy.     

Developmental changes in marmoset granulosa cell responsiveness to insulin-like growth factor-I: interactions with follicle-stimulating hormone and estradiol

The biological actions of insulin-like growth factor-I (IGF-I) on granulosa cell steroidogenesis at defined stages of preovulatory follicular development in the marmoset monkey were examined. Studies were carried out by primary cell culture of granulosa cells derived from small antral (0.5-1.mm diameter) and large preovulatory (2-3.mm diameter) follicles collected during the mid-late follicular phase of the ovarian cycle. IGF-I (0.3-100 ng/ml) had no effect on progesterone accumulation or aromatase activity during 48-h culture of granulosa cells from small follicles. Progesterone accumulation by cells from large follicles was also unaffected by IGF-I over the same time period, although aromatase activity was stimulated in a dose-dependent manner (18-fold increase over basal levels with a maximally stimulatory dose of 30 ng IGF-I/ml). In contrast, granulosa cells from small and large follicles responded to IGF-I in terms of both progesterone accumulation and aromatase activity after longer periods of culture (4 days for progesterone; 6 days for aromatase). Concurrent treatment of granulosa cells from small follicles with estradiol (10(-7) M) enhanced the dose-dependent actions of IGF-I on both indices of steroidogenesis and advanced the time at which IGF-I stimulated activity was first detectable. The effects of estradiol on granulosa cell IGF-I responsiveness were independent of cell number. A synergistic action of IGF-I on FSH-stimulated granulosa cell steroidogenesis was not apparent.(ABSTRACT TRUNCATED AT 250 WORDS)