Observation of the Top Quark.
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Biomedical subjects
Publications and source records attributed to J Borders.
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The p53 tumor suppressor gene has been found to be altered in almost all human solid tumors, whereas K-ras gene mutations have been observed in a limited number of human cancers (adenocarcinoma of colon, pancreas, and lung). Studies of mutational inactivation for both genes in the same patient's sample on non-small-cell lung cancer have been limited. In an effort to perform such an analysis, we developed and compared methods (for the mutational detection of p53 and K-ras gene) that represent a modified and universal protocol, in terms of DNA extraction, polymerase chain reaction (PCR) amplification, and nonradioisotopic PCR-single-strand conformation polymorphism (PCR-SSCP) analysis, which is readily applicable to either formalin-fixed, paraffin-embedded tissues or frozen tumor specimens. We applied this method to the evaluation of p53 (exons 5-8) and K-ras (codon 12 and 13) gene mutations in 55 cases of non-small-cell lung cancer. The mutational status in the p53 gene was evaluated by radioisotopic PCR-SSCP and compared with PCR-SSCP utilizing our standardized nonradioisotopic detection system using a single 6-microns tissue section. The mutational patterns observed by PCR-SSCP were subsequently confirmed by PCR-DNA sequencing. The mutational status in the K-ras gene was similarly evaluated by PCR-SSCP, and the specific mutation was confirmed by Southern slot-blot hybridization using 32P-labeled sequence-specific oligonucleotide probes for codons 12 and 13. Mutational changes in K-ras (codon 12) were found in 10 of 55 (18%) of non-small-cell lung cancers. Whereas adenocarcinoma showed K-ras mutation in 33% of the cases at codon 12, only one mutation was found at codon 13. As expected, squamous cell carcinoma samples (25 cases) did not show K-ras mutations. Mutations at exons 5-8 of the p53 gene were documented in 19 of 55 (34.5%) cases. Ten of the 19 mutations were single nucleotide point mutations, leading to amino acid substitution. Six showed insertional mutation, and three showed deletion mutations. Only three samples showed mutations of both K-ras and p53 genes. We conclude that although K-ras and p53 gene mutations are frequent in non-small-cell lung cancer, mutations of both genes in the same patient's samples are not common. We also conclude that this universal nonradioisotopic method is superior to other similar methods and is readily applicable to the rapid screening of large numbers of formalin-fixed, paraffin-embedded or frozen samples for the mutational analysis of multiple genes.
PURPOSE: We evaluated the accuracy of transcranial Doppler (TCD) and magnetic resonance angiography (MRA) as a screening modality for cerebral infarct in patients with sickle cell disease. PATIENTS AND METHODS: Twenty-four patients with sickle cell disease were assessed for a cerebral infarct. Each patient underwent neurologic examination, psychometric evaluation, magnetic resonance imaging (MRI), MRA, and TCD. Presence of a cerebral infarct was determined by an MRI and supported by neurologic and neuropsychometric evaluation. All investigators were unaware of the patient's status. Cerebral vasculature by MRA was categorized as being occluded, stenotic, or normal. The maximum TCD velocity of the right and left middle cerebral artery were assessed. Various cutoffs were used to determine the sensitivity and specificity of TCD. RESULTS: MRA had a sensitivity and specificity of 100% and 92%, respectively. At a maximum velocity cutoff of 180 cm/s, the sensitivity and specificity of TCD were 20% and 67%, respectively. No maximum velocity on TCD produced both sensitivity and specificity > 50%. CONCLUSION: MRA is more sensitive than TCD when the middle cerebral artery for maximum velocity is compared. Prospective evaluation is warranted to determine if TCD is useful as a screening technique for cerebral infarction in children with SCD.
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Quantitative dual-energy radiographic absorptiometry (DRA) and dual-photon absorptiometry (DPA) were compared to determine the best means of assessing bone density. Both methods were used to evaluate the lumbar spine in 107 women (aged 35-84 years [mean, 64 years]) referred for evaluation of osteoporosis risk. High correlation was documented between measurements derived by the two techniques, with a .95 linear regression coefficient for the total spine density measurement. Age-related regression equations were similar in slope but manifested different intercepts. Bone mineral density values derived with DRA were consistently lower than those obtained with DPA (conversion equation: DPA density = [1.067 X DRA density] + 0.163). Besides the inherent imperfections of each system, it was found that inaccurate identification of intervertebral spaces on the low-resolution DPA images introduced errors in patient data. DRA may replace DPA as the dedicated projectional densitometric procedure of choice for technical reasons, but at present a conversion equation must be used to compare DRA data to DPA data.