Autolytic activation of recombinant human 72 kilodalton type IV collagenase.

Human 72 kDa type IV collagenase (gelatinase A, MMP-2) was expressed in a baculovirus/insect cell system. The enzyme was produced in the wild-type form and in two mutant forms, where the active site Glu375 was substituted by Asp or Gln. The mutated proteins had strongly reduced or no detectable activity, respectively, allowing detailed analysis of rapid autoactivation reactions.

[Clinical decision aids using ultrasound biomicroscopy].

Ultrasound biomicroscopy (UBM) allows high-resolution imaging of the anterior segment due to ultrasound transducers between 50 and 100 MHz. At present stage of experience, mainly analysis of iris and ciliary body structures seem to be clinically rewarding. This is demonstrated by two similar looking tumors, involving the chamber angle.

Localization of proteins HL29 and HL31 from Haloarcula marismortui within the 50 S ribosomal subunit by chemical crosslinking.

Isolated 50 S ribosomal subunits from the halophilic archaebacterium Haloarcula marismortui were treated in situ with the homobifunctional and cleavable crosslinking reagent dithiobis(succinimidyl propionate) (12 A). Several crosslinked complexes were obtained. Among these were the protein pairs HmaL4-HL29 and HmaL18-HL31; HL29 and HL31 are ribosomal proteins without any equivalent in eubacterial ribosomes.

HL35e and HLA: primary structure of two very basic and cysteine-rich ribosomal proteins from Haloarcula marismortui.

Two small and very basic ribosomal proteins have been purified from the 50S ribosomal subunit of the archaebacterium Haloarcula marismortui by RP-HPLC. The complete primary structures of these two proteins, which we refer to as HL35e and HLA, have been determined by protein chemical methods. Both proteins are characterized by a high content of basic amino acids and the presence of two pairs of cysteines in each polypeptide chain, one of which resembles the C4-zinc-finger motif.

N-terminal modification and amino-acid sequence of the ribosomal protein HmaS7 from Haloarcula marismortui and homology studies to other ribosomal proteins.

The ribosomal protein HmaS7 from the 30S subunit of the extreme halophilic archaeum Haloarcula marismortui was isolated by semi-preparative RP-HPLC. The complete amino-acid sequence of this protein was determined by automated microsequence analysis of appropriate peptide fragments from several proteinase digests. The entire protein consists of 205 amino acids with a corresponding molecular mass of 22580 Da.

Identification of cross-linked amino acids in the protein pair HmaL23-HmaL29 from the 50S ribosomal subunit of the archaebacterium Haloarcula marismortui.

50S ribosomal subunits from the extreme halophilic archaebacterium Haloarcula marismortui were treated with the homobifunctional protein-protein cross-linking reagents diepoxybutane (4 A) and dithiobis(succinimidyl propionate) (12 A). The dominant product with both cross-linking reagents was identified on the protein level as HmaL23-HmaL29, which is homologous to the protein pair L23-L29 from Escherichia coli [Walleczek, J., Martin, T.

Fluidity of the microsomal membrane and cytochrome P450 reduction kinetics of pig liver microsomes as a consequence of organic solvent impact.

1. The effect of the aromatic solvents toluene, xylene and ethylbenzene on microsomal membrane fluidity and anaerobic NADPH-reduction kinetics were studied. 2.

Induction of inflammatory mediator release (serotonin and 12-hydroxyeicosatetraenoic acid) from human platelets by Pseudomonas aeruginosa glycolipid.

Purified glycolipid from Pseudomonas aeruginosa induced the generation of significant amounts of 12-hydroxyeicosatetraenoic acid (12-HETE) and serotonin release from human platelets. The release of serotonin was first observed 2 min after addition of the glycolipid and increased with time. Significant serotonin release was obtained at glycolipid concentrations above 5 micrograms/ml and increased dose-dependently up to 100% at glycolipid concentrations above 40 micrograms/ml.

Interleukin 6--a potential mediator of lethal sepsis after major thermal trauma: evidence for increased IL-6 production by peripheral blood mononuclear cells.

We studied the plasma levels of the acute phase mediator interleukin 6 (IL-6) in 21 severely burned patients (burn injuries ranging from 24% to 75% total body surface area). The posttraumatic course of the IL-6 plasma levels was closely related to the clinical outcome. The nonseptic survivors as well as survivors with suspected sepsis (n = 14) exhibited maximal amounts of IL-6 (251 +/- 32 pg/mL) during the first 3 days post-burn, which subsequently returned to values within the normal range (days 30 to 50; 26 +/- 8 pg/mL).

Impairment of specific host defense mechanisms in patients with chronic post-traumatic osteomyelitis.

We studied both in vivo and in vitro specific host defense mechanisms in patients suffering from chronic post-traumatic osteomyelitis (n = 26). The cell-mediated immunity in vivo was impaired as indicated by the reduced reactivity in the delayed type hypersensitivity skin test. The concanavalin A or phytohemagglutinin-induced T-cell proliferation in vitro was markedly decreased in comparison to healthy donors.

Basophil releasability in severely burned patients.

Thermal injury is known to induce dysregulation of the immune system; however, the precise mechanisms have to be clarified. We investigated the histamine release of basophil granulocytes from severely burned patients (n = 12) after stimulation with anti-IgE or the Ca-ionophore A 23187, respectively. The anti-IgE-induced basophil histamine release of all patients was reduced in comparison to healthy donors beginning at day one postburn (p.

Evidence for an additional archaebacterial gene cluster in Halobacterium marismortui encoding ribosomal proteins HL46e and HL30.

A small and extremely basic ribosomal protein (HL46e) has been purified from Halobacterium marismortui using reversed-phase high-performance liquid chromatography (HPLC). The amino acid sequence of the protein was determined by automated N-terminal and internal sequence analysis. Comparison of this sequence with other ribosomal protein sequences from eubacteria, archaebacteria and eukaryotes revealed a strong homology to SL46e from Sulfolobus solfataricus, YeaL46 from yeast and RL39 from rat.

Use of a hapten specific anti-dansyl antibody for the localization of ribosomal proteins by immuno electron microscopy.

The fluorescent reagent dansyl chloride has been used as an immunological marker for the electron microscopic localization of ribosomal proteins on the surface of 50S ribosomal subunits. The proteins BstL1 from Bacillus stearothermophilus and EcoL1 from Escherichia coli were dansylated to various degrees and reconstituted into the L1-deficient E. coli 50S subunits from mutant MV17-10.

Induction of inflammatory mediators (histamine and leukotrienes) from rat peritoneal mast cells and human granulocytes by Pseudomonas aeruginosa strains from burn patients.

Clinical isolates of Pseudomonas aeruginosa from severely burned patients were analyzed with regard to their capacity to induce inflammatory-mediator release from rat mast cells or human granulocytes. The bacterial strains were characterized according to their cell-associated hemolysin activity as well as their secreted hemolysin and phospholipase C activities. P.

[Cardiac tamponade following open-heart surgery (author's transl)].

Cardiac tamponade occurred in 9 (1,8%) of 488 patients following open-heart surgery. Delayed cardiac tamponade was observed in one case (0,2%). The mediastinal tamponade was caused by early occlusion of the chest tubes, pericardium closure and administration of anticoagulants.

Focus path of a laser beam deflected by a prismatic polygon mirror: its calculation and optimization.

The focus path of a laser beam deflected by a prismatic polygon mirror was calculated. It is shown that the larger the focal distance of the objective or the polygon radius, the closer the focus path approaches a straight scanning line. Especially in the case of small pattern scanning, a remarkably small deviation between actual focus path and straight scanning line can be achieved by suitable sizing and arrangement of the optical elements.

[Oncogen infectious nucleoprotein and herpes virus hominis type 2 in stages of cervical cancer. Investigatious by indirect immunofluorescens].

Using HVH 2 antisera of rabbit (titre 1:256, dilution 1:10), antologous serum of cervix carcinoma patients (dilution 1:1 to 1:5) and allogenic serum of cervix carcinoma patients without HVH 2 antibodies (dilution 1:1 to 1:5), tumour material obtained from various cancerization stages of cervix carcinoma in 50 cases was incubated following trypsination to single cells with fluoresceine isothiocyanate labelled anti-human globulin and anti-rabbit globulin. Apart from HVH antigens significantly differing in quantity from one stage to the other as detected by immune fluorescence methods, a perinuclear distinct antigen, unrelated to HVH antigens, has also been demonstrated. The last mentioned one might be a specific tumour antigen or an embryonic antigen.