MMP-9 activation by tumor trypsin-2 enhances in vivo invasion of human tongue carcinoma cells.

Various human cancer cells express tumor-associated trypsinogen-2 (TAT-2), which can efficiently activate matrix metalloproteinases (MMPs) in vitro. MMP-2 and MMP-9 are particularly associated with the invasive malignant potential of several tumors. To investigate the role of TAT-2 in tumor invasion, we overexpressed TAT-2 in two malignant human squamous cell carcinoma cell lines of tongue and in non-malignant human papilloma virus transformed gingival keratinocytes. The TAT-2 overexpression significantly increased the levels of active MMP-9 in the most malignant cell line. TAT-2-transfected cells intravasated (invaded blood vessels) up to 60% more efficiently than did the control cells in an in vivo chick embryo chorioallantoic membrane invasion model. This increased intravasation was almost completely abolished by a specific tumor-associated trypsin inhibitor (TATI). These results indicate that TAT-2 has a role in the invasive growth of tumors, either alone or in cascade with gelatinases, especially by generating active MMP-9.

Expression and regulation of collagenase-2 (MMP-8) in head and neck squamous cell carcinomas.

MMP-8 (collagenase-2) is the most effective collagenase to initiate type I collagen degradation. Since initiation of lysis of the surrounding collagen matrix is an essential prerequisite for carcinoma cells to spread, this study investigated the expression of MMP-8 in squamous cell carcinoma (SCC) of the head and neck in vivo and in vitro. Most of the recently established head and neck carcinoma cell lines (22/25), corresponding tumour (5/7) and dermal (2/2) fibroblasts, commercial tongue carcinoma (HSC-3 and SCC-25), and transformed keratinocyte cell lines of the tongue (IHGK) and skin (HaCaT) expressed MMP-8 mRNA analysed by the PCR method. Western blotting revealed a latent 50 kD band in concentrated culture media of carcinoma cells and corresponding tumour and dermal fibroblasts. The expression of immunoreactive MMP-8 protein was reduced 30% by transforming growth factor beta-1 (TGF-beta1) at 1 ng/ml concentration and 60% at 10 ng/ml concentration, but up-regulated 2- and 2.5-fold after 10 nM and 100 nM phorbol 12-myristate 13 acetate (PMA), respectively. Immunohistological staining localized MMP-8 protein in a few malignant invading tumour cell islands, certain fibroblasts, polymorphonuclear neutrophils (PMNs), and plasma cells. In situ hybridization revealed a faint sporadic signal in carcinoma cells of all eight tissue sections analysed. It is concluded that tissue from head and neck carcinomas can express MMP-8 both in vivo and in vitro. Since the amount of MMP-8 in carcinoma and stromal cells is rather low, MMP-8 may have a potential role, with other collagenases, in the proteolysis of connective tissue associated with the spreading of invasive carcinoma.

Estimation of individual dust exposure by magnetopneumography in stainless steel production.

The objectives of the study were to measure the magnetic dust lung burden of workers in stainless steel production by magnetopneumography (MPG) and to investigate the relationship of the results with air-borne concentrations of dust, total and hexavalent chromium as well as urinary excretion of chromium. There were 128 workers from the chromite mine, sintering plant, ferrochrome smelter, stainless steel smelting shop, cold rolling mill and welding shop in the exposed groups and five persons from the office staff in the control group. The remanent magnetic field (RMF) in the lungs was slightly elevated among workers in the ferrochromium and steel smelting shops; the levels were, however, lower than those reported for welders earlier and those observed in the welding/repair shop. Workers in the mine, concentrator and sintering plants and in the cold rolling mill exhibited remanent magnetic fields comparable to the referents. There was a relationship between the RMF and the actual urinary chromium concentration. Miners and concentrator and sintering plant workers showed retarded relaxation rate (ReR) of the remanent magnetic field. However, the RMF of the first two of these groups were low (< 0.1 nT) and this made it difficult to measure the ReR accurately. The duration of exposure correlated weakly but significantly with the relaxation rate, while smoking was not related to it.

Pharmacokinetics of enrofloxacin in newborn and one-week-old calves.

The pharmacokinetic behaviour of enrofloxacin was compared in four one-day-old and four one-week-old calves in order to find out if there were any age-related differences. Mean volume of distribution (Vd(ss)) and clearance (Cl) were significantly smaller in newborn calves: Vd(ss) was 1.8 and 2.3 L/kg, while clearance was 0.19 and 0.39 L/kg.h in newborn and one-week-old calves, respectively. Mean elimination half-life (t1/2 beta) did not differ significantly in newborn and in one-week-old calves: mean t1/2 beta was 6.6 h and 4.9 h, respectively. Enrofloxacin was metabolized to ciprofloxacin both by newborn and one-week-old calves, but the maximum concentration (Cmax) of ciprofloxacin was lower and the time when maximum concentration was reached (tmax) later in newborn calves. We conclude that the dosage of enrofloxacin should be adjusted according to age when administered to very young calves.

1H and 13C NMR analysis of some trichothecenes.

1H and 13C NMR chemical shifts were determined from the NMR spectra of low concentration solutions of T-2 toxin, T-2 triol, HT-2 toxin, diacetoxyscirpenol, and neosolaniol. Isomerization of neosolaniol was observed to occur in chloroform solution. 1D and 2D 1H and 13C NMR techniques were used in order to determine the structure of the isomerization product, finally identified as 4 beta, 8 alpha-diacetoxy-3 alpha, 15-dihydroxy-12,13-epoxytrichothec-9-ene.

Measurement of lung-retained contaminants in vivo among workers exposed to metal aerosols.

In Finland the amount of lung-retained contaminants has been measured among welders, foundry workers, and iron and steel factory workers. About 300 subjects have so far been measured since 1976. The method of measuring is based on the magnetic properties of metal aerosols. At the very beginning the method resembled the one suggested by D. Cohen. The original method was too slow, and data processing was not suitable for clinical or hygienic work. Therefore, the automatic and computerized instrument for measuring lung-retained contaminants was developed in cooperation with Outokumpu Oy. The resolution of the magnetic measurement is better than 0.05 nT corresponding to a sensitivity of 0.5 mg of magnetic contaminants in the lung in the form of magnetite. These figures mean that, in practice, a shipyard welder has enough contamination for measurement after one month's exposure. In addition, the present paper reviews the magnetic properties of metal aerosols briefly and summarizes the results of measuring different occupational groups.

The specificity and multiplicity of human placental xenobiotic-metabolizing monooxygenase system studied by potential substrates, inhibitors and gel electrophoresis.

The specificity of the placental monooxygenase system to metabolize foreign compounds was studied by using different potential substrates and inhibitors and by performing electrophoresis of placental microsomes. Placental preparations from smokers catalyzed benzo(a)pyrene hydroxylation, 7-ethoxycoumarin O-deethylation and 2,5-diphenyloxazole hydroxylation, but not biphenyl hydroxylation at 2-, 3- or 4-carbon, aldrin epoxidation to dieldrin or coumarin hydroxylation or aminopyrine N-demethylation. Enzyme activities were inhibited by alpha-naphthoflavone, but to a much lesser extent by SKF 525-A or metyrapone. Correlations between the metabolism of benzo(a)pyrene, 7-ethoxycoumarin and 2,5-diphenyloxazole were highly significant. There was a clear difference in Michaelis-Menten constant of 7-ethoxycoumarin O-deethylation between placentas from smokers and nonsmokers. Gel electrophoresis revealed that protein bands of placental microsomes in the region of cytochrome P-450 enzymes were less prominent than those of rat liver microsomes, a finding that accorded with the relative amounts of cytochrome P-450. There were no consistent differences in the electrophoretic pattern between placentas of variable benzo(a)pyrene hydroxylase activities. Results show that the human placental monooxygenase system is restricted in substrate specificity, that there may be a qualitative difference between smokers and nonsmokers and that the increase in several enzyme activities by cigarette smoking cannot be detected by the standard gel electrophoresis.