Molecular cloning of human synovial cell collagenase and selection of a single gene from genomic DNA.

We used a subclone of a rabbit genomic clone for collagenase that cross-hybridizes with human synovial cell messenger RNA (mRNA) to identify a human collagenase complementary DNA (cDNA) clone. The human cDNA clone is 2.1 kilobases (kb) and selects a mRNA transcript of approximately the same size from primary cultures of rheumatoid synovial cells that produce collagenase, but no mRNA is selected from control (nonproducing) synovial fibroblasts. Restriction enzyme analysis and DNA sequence data indicate that our cDNA clone is full length and that it is identical to that recently described for human skin fibroblast collagenase. The cDNA clone identified a single collagenase gene of approximately 17 kb from blots of human genomic DNA. The identity of human skin and synovial cell collagenase and the ubiquity of this enzyme and of its substrates, the interstitial collagens types I, II, and III, imply that common mechanisms controlling collagenolysis throughout the human body may be operative in both normal and disease states.

Molecular cloning, sequence analysis, and functional expression of a novel growth regulator, oncostatin M.

Oncostatin M is a polypeptide of Mr approximately 28,000 that acts as a growth regulator for many cultured mammalian cells. We report the cDNA and genomic cloning, sequence analysis, and functional expression in heterologous cells of oncostatin M. cDNA clones were isolated from mRNA of U937 cells that had been induced to differentiate into macrophagelike cells by treatment with phorbol 12-myristate 13-acetate, and a genomic clone was also isolated from human brain DNA. Sequence analysis of these clones established the 1,814-base-pair cDNA sequence as well as exon boundaries. This sequence predicted that oncostatin M is synthesized as a 252-amino-acid polypeptide, with a 25-residue hydrophobic sequence resembling a signal peptide at the N terminus. The predicted oncostatin M amino acid sequence shared no homology with other known proteins, but the sequence of the 3' noncoding region of the cDNA contained an A + T-rich stretch with sequence motifs found in the 3' untranslated regions of many cytokine and lymphokine cDNAs. Oncostatin M mRNA of approximately 2 kilobase pairs was detected in phorbol 12-myristate 13-acetate-treated U937 cells and in activated human T cells. Transfection of cDNA encoding the oncostatin M precursor into COS cells resulted in the secretion of proteins with the structural and functional properties of oncostatin M. The unique amino acid sequence, expression by lymphoid cells, and growth-regulatory activities of oncostatin M suggest that it is a novel cytokine.

Experience with 40,000 Papanicolaou smears.

Forty thousand consecutive cytologic smears and subsequent diagnostic procedures resulted in the diagnosis of 41 carcinomas in situ, 35 microinvasive and invasive carcinomas, and 24 severe dysplasias for a yield of significant neoplasia of one lesion per 400 Papanicolaou smears. Twenty-five of the carcinomas in situ and microinvasive and invasive carcinomas were diagnosed in patients with atypical smears indicating that all patients with persistent atypical smears require evaluation by tissue examination. Seventy-eight percent of the 119 patients subjected to conization either had carcinoma in situ, microinvasive and invasive carcinoma, or significant cervical dysplasia. Post-operative complications following conization were negligible. In addition there were no postconization deleterious effects on three concurrent and nine subsequent pregnancies. A history of gonorrhea places a patient at a higher risk of developing cervical carcinoma. Annual performance of cytologic smear evaluation is indicated in all sexually active women and in all virginal women over 20 years of age.

Causes of trauma in broilers arriving dead at poultry processing plants.

A post mortem examination was made on 1324 broiler carcases which arrived dead after being transported to six processing plants. The carcases were dissected and evaluated for trauma and obvious cause of death. Of the birds sent to the plants on the day of collection 0.19 per cent arrived dead and 35 per cent of these were thought to have died as a result of trauma. Haemorrhage associated with dislocation of the femur at the hip was the most common cause of trauma. Ruptured livers and crushed heads were other important traumatic injuries.

Carcinoma of the cervix in oral contraceptive steroid and IUD users and nonusers.

A total of 40,211 cytologic examinations and subsequent diagnostic procedures resulted in the diagnosis of carcinoma of the cervix in 76 patients. The 76 patients did not demonstrate any variations in use of oral contraceptive steroids or intrauterine devices when compared to 780 randomly selected control patients. This study is in agreement with our earlier report demonstrating no increased risk of carcinoma of the cervix in oral contraceptive steroid users compared to nonusers.

Effect of catching method on the prevalence of broken bones in end of lay hens.

Five trials were conducted in which six methods of picking end of lay hens from battery cages were compared with three people performing the picking. Pulling the birds over a breast support slide, which created a smooth surface over the feeding trough, did not affect the prevalence of broken bones. In one trial, picking birds by one leg resulted in about three times more fractured bones than picking by two legs. This effect, however, was not evident in three other trials, suggesting that the advantage from two leg picking would only exist with some pickers or some flocks. Pulling more than one bird at a time by one leg through the cage entrance was associated with more skeletal damage than pulling birds individually by two legs. Picking the birds by one leg was easier to perform than picking by two legs.

The depletion of glycogen stores and indices of dehydration in transported broilers.

Broilers were either not transported or were transported for 2, 4 or 6 hours after having been subjected to food withdrawal times of less than one hour or of ten hours. The birds were then slaughtered using normal commercial practices. The longer period of food deprivation reduced liver weight and glycogen content, and circulating glucose concentrations. It also elevated the ultimate pH value (pHu) in the biceps muscle and by implication, therefore, reduced its glycogen content. With longer journey times, liver weight and glycogen content decreased. Transport had an inconsistent effect on glycogen concentration in the pectoral muscle but progressively reduced its pHu. In contrast, pHu in the biceps progressively increased, by implication because transport depleted muscle glycogen levels. Birds transported further had higher concentrations of total protein in their plasma which, though this was not significant, also had a higher osmolality. This suggests that transported birds became dehydrated. Additionally, the depletion of body glycogen stores might be associated with the perception of fatigue.

Homology between exon-containing portions of rabbit genomic clones for synovial cell collagenase and human foreskin and synovial cell mRNAs.

Portions of rabbit genomic clones containing exons cross-hybridize with human synovial cell and human foreskin fibroblast mRNAs. These cross-hybridizing genomic fragments have been subcloned into pBR328 and may be useful probes in isolating cDNA and genomic clones from human tissue, and perhaps from other species as well. In addition, DNA sequence data on a 530 bp cDNA clone for rabbit synovial-cell collagenase indicate that this clone is composed primarily of 3'-untranslated region. As such, it is probably not useful in cross-species hybridizations.