Detection of human papilloma virus (hpv) dna in genital biopsy specimens by in situ hybridization with biotinylated probes

The presence of human papilloma virus (HPV) nucleotide sequences in paraffin sections of genital biopsies was examined by in-situ hybridization using non-isotopic, random primer biotin labeled probes representing HPV type 16. Biotin labeling of probes was performed using DNA labeling and commercially provided phenotype detection kit. Hybridization was performed under stringent conditions. The hybrids were detected by using Streptavidin and biotinylated alkaline phosphatase and visualized with an enzyme catalyzed chemiluminescent detection reaction. In-situ hybridization with biotinylated probes was a useful technique for the identification of HPV infection. The results were compared with the results obtained with dot blot hybridization probes. The background with biotinylated probes was very low. Using non radioactive robes the localization of hybrids at the cellular level was not better than dot blot labeled probes (AU)

Effect of heat conservation during and after major abdominal surgery on muscle protein breakdown in elderly patients.

Changes in mean body temperature and muscle protein metabolism were studied in elderly patients undergoing large bowel surgery. Two groups were studied: in one, efforts were made to maintain the patients normothermic during and after surgery by warming the fresh gases, the i.v. fluids, by placing warmed cotton padding around the exposed parts of the body and by covering the patients with a metallized plastic sheet in the recovery period. The other group received routine management. Otherwise the anaesthetic technique was comparable. The excretion of the amino acid 3-methylhistidine (3-MeH), an indicator of muscle protein breakdown, and urea nitrogen loss were measured in the urine collected the day before, and on the 2nd and 4th postoperative days. Prevention of heat loss during and after surgery caused a significant decrease in muscle protein degradation and nitrogen loss.

Separation and quantitation of oxypurines by isocratic high-pressure liquid chromatography: application to xanthinuria and the Lesch-Nyhan syndrome.

An isocratic HPLC technique has been developed for the separation and measurement of urine and plasma oxypurines in a patient with xanthinuria. The case history and laboratory data are presented. Xanthine excretion was 172 mg/g creatinine and hypoxanthine was 45 mg/g creatinine. Uric acid was too small to be measured but uricase determination showed only 3 mg/24 hr. Serum oxypurine analysis showed hypoxanthine 0.87 mg/dl and xanthine 0.35 mg/dl. Uric acid was not seen in this patient's serum but could be readily measured in normal control subjects. The technique can also be used to separate nucleotides from purine bases, and we have demonstrated its application to the measurement of erythrocyte hypoxanthine guanine phosphoribosyl transferase and adenine phosphoribosyl transferase in a kindred associated with the Lesch-Nyhan syndrome.

Comparison of CK-MB fractionation by antibody inhibition reaction and DEAE-sephadex chromatography.

1. CK-MB isoenzyme fractionation has been compared between DEAE-sephadex chromatography and the CK-M-subunit inhibiting antibody procedure. 2. It was found that the inhibiting antibody method generally gave lower values than the chromatography procedure. 3. The above difference can be partially attributed to inhibitory factors in the antibody system. 4. It was concluded that although there was general agreement between the two procedures, the CK-M-subunit inhibiting antibody procedure should be viewed with caution at low creatine kinase activities, and should never be done without the appropriate reagent blanks.

Uric acid estimation: a comparison of the manual uricase-UV and the phosphotungstate auto-analyzer methods.

1. Uric acid estimations by the uricase-UV and the autoanalyzer colorimetric procedures were compared. 2. At all levels of serum uric acid concentration measured, results with the uricase-UV method were significantly lower than those with the phosphotungstate method. 3. The data indicate that the critical uric acid level (measured by the autoanalyzer) at which allopurinol, uricosuric, and other agents are instituted should be decreased approximately 10 per cent when determinations are done by the uricase method.