A method for isolation of sequences missing in one of two related genomes

We describe a novel technique for isolation of sequences that are present in one genome (tracer), but absent in another (driver). Tracer DNA, cleaved with Sau 3A and capped with a single stranded PCR adapter, is allowed to hybridize with an excess of sheared biotinylated driver; biotinylated DNA and its hybrids with the tracer are removed by phenol/chloroform extraction after incubation with streptavidin. After several rounds of subtraction the ends of self-annealed tracer molecules from the nonextractable fraction are filled-in with Tag polymerase and amplified, using the single stranded PCR adapter as a primer. The method has been applied to purification of fragments from a 2.9 kb plasmid added to E. coli DNA at equimolar quantity. Plasmid derived fragments (250-1000 bp), initially comprising 1/1400th part of tracer DNA, were purified to homogeneity after two rounds of subtraction followed by PCR.     

An antinociceptive effect of the intraperitoneal injection of nifedipine in rats, measured by tail-flick test

The tail-flick (TF) technique was used to assess the antinociceptive properties of nifedipine (NIF) given intraperitoneally (i.p.). First, the most suitable intensity of the noxious stimulus (temperature of the bulb) has been ascertained and used in the main study. Male Sprague-Dawley rats received NIF, dissolved in dimethyl sulfoxide (DMSO) at the doses of 0.0, 0.5, 2, 5, 10 and 15 mg/kg, or control with no injection. For the main study, the noxious stimulus was limited to 15 sec (cut-off time) and TF latencies were recorded up to 120 min. The antinociceptive response was expressed as the area under the curve for each rat and analyzed by one-way ANOVA. The antinociceptive response to the lower doses of NIF (0.5 and 2 mg/kg) did not differ from control (no injection) and DMSO alone. Significance was found at 5, 10 and 15 mg NIF with no difference among the doses. However, there was an increasing tendency of the mean values from 0.5 to 15 mg NIF resulting in a positive correlation. The correlation coefficient was 0.32483 (p = 0.015) and regression equation Y = (19.37) x dose + 1320. Our data suggest that spinal mechanisms are involved in NIF-induced antinociception.     

Influence of dietary iron deficiency on acute metal intoxication

The frequent difficulties encountered in the diagnosis of pediatric sarcomas, caused by the lack of observable differentiation at the light microscopic level, has led to the routine use of immunohistochemistry in pediatric surgical pathology. To a large degree the advent of this staining technique has led to the correct assessment of many perplexing lesions that previously would have been given inconclusive diagnoses. However, with increased usage and testing, it has become apparent that there are few, if any, "magic bullets" in immunohistochemistry for pediatric pathologists. Thus, it behooves diagnosticians to be careful in the usage of this technique, to be aware of possible discrepancies in its results, and to remember the ancillary nature of its application. The following article will review selected markers commonly used in pediatric surgical pathology, from both previous reports and the author's perspective, and will briefly consider several new phenotypic markers which have potential utility with childhood sarcomas.     

Some problems of the moral and psychological training of medical service personnel

The authors' experience with 45 papillosphincterotomies, performed for diseases of bile passages and chronic pancreatitis, is reported. The details of the operative technic, employed by the authors, are described. The technic of papillosphincterotomy on a conus-shape sound by means of a "laser scalpel" proved to be the most perfect one. Careful observation of the surgical technic made it possible to gain favourable results--only 1 of 45 operated patients died. Papillosphincterotomy is largely advocated in stenoses and concretions in the large duodenal papilla, strictures of choledocho-duodenoanastomoses, care should be taken not to overwiden indications to the operation concerned.     

Report from the Maryland Epidemiology Schizophrenia Linkage Study: no evidence for linkage between schizophrenia and a number of candidate and other genomic regions using a complex dominant model

Our collaborative group has undertaken a linkage study of schizophrenia, using a systematic sample of patients admitted to Maryland hospitals. An initial sample of 39 families, each having two or more affecteds, was available for genotyping candidate genes, candidate regions, and highly polymorphic markers randomly distributed throughout the genome. We used a single complex dominant model (with a disease gene frequency of 0.005 and age-dependent penetrance for affected phenotype: for under 35, penetrance = .45; for 35 and older, penetrance = .85). We report here 130 markers, which met the exclusion criteria of LOD score < -2.00 at theta > 0.01 in at least 10 informative families, and no evidence for heterogeneity. We also report here markers that were tested as candidates for linkage to the schizophrenic phenotype. They were selected based on the following criteria: a) proximity to reported chromosomal rearrangements (both 5q and 11q), b) suggestions of linkage from other families (5q), or c) presence of a candidate gene (5q, 11q, 3q: Dopamine receptors 1, 2, and 3, respectively). We also tested for mutations of codon 717 in exon 17 of the amyloid precursor protein (APP) gene and were unable to detect the C to T substitution in our schizophrenic group.