The fibrin-binding site of human plasminogen. Arginines 32 and 34 are essential for fibrin affinity of the kringle 1 domain

Kringle 1 (Tyr 79/Leu 80-His 167 and Tyr 79/Leu 80-Tyr 173), a chymotryptic fragment of human plasminogen that has high affinity for fibrin and omega-aminocarboxylic acids, has been subjected to modification with 1,2-cyclohexanedione to identify arginine residues essential for ligand binding. Reaction of 1,2-cyclohexanedione with kringle 1 was found to rapidly abolish the fibrin-Sepharose affinity of the fragment, whereas the affinity for lysine-Sepharose was lost at a significantly slower rate. Successive affinity chromatography of modified kringle 1 on fibrin- and lysine-Sepharose was used to separate kringle 1 that lost affinity for fibrin-, but retained affinity for lysine-Sepharose from kringle 1 that lost affinity for both affinants. The modified proteins were subjected to structural studies in order to locate the labeled arginine residues in kringle 1. These studies have revealed that modification of Arg 34 leads to the loss of both the fibrin- and lysine-Sepharose affinities of kringle 1, whereas reaction of Arg 32 abolishes fibrin affinity but leaves lysine-Sepharose affinity unaltered. The results suggest that Arg 32 and Arg 34 are both involved in fibrin binding and that Arg 34 is also involved in binding omega-aminocarboxylic acids. Previous NMR studies on kringles have indeed shown that the segment containing residue 34 is in the proximity of and interacts with the omega-aminocarboxylic acid-binding site. This interaction may explain the influence of omega-aminocarboxylic acids on fibrin binding by kringle 1.     

Administration of antibiotics in combination. The antibacterial activity of blood serum and urine after a simultaneous administration of aminoglycosides and ampicillin

Blood serum and urine samples collected from a group of volunteers treated with single doses of ampicillin and aminoglycoside preparations given separately or in combination were tested for their antimicrobial activity against the reference strains Staphylococcus aureus SZK 76/69 and ATCC 6538, Pseudomonas aeruginosa SZK 444 and SZK 385, and Escherichia coli SZK 326/71. Out of all antimicrobials and their combinations tested the most powerful was the combination of netilmicin with ampicillin. Of the therapeutic combinations used nowadays in clinical practice the combined use of gentamicin and ampicillin proved also effective. These antibiotic combinations appear thus to be best suited for the treatment of mixed Pseudomonas aeruginosa and Staphylococcus aureus infections and of urinary tract infections caused by bacterial strains exhibiting in the in vitro susceptibility assays a reduced sensitivity to some of the antibiotic preparations used.     

Urinary excretion of mandelic and phenylglyoxylic acids after human exposure to styrene vapour

The kinetics of the urinary excretion of mandelic and phenylglyoxylic acids were studied in volunteers exposed to the known concentrations of styrene vapour. The level and the time of exposure were suitably changed to simulate situations in the industrial environment. The aim was to find out the reasons for the contradictory reports in the literature and to verify parameters characterizing the course of excretion of both metabolites. It was found that the course of mandelic acid excretion might be influenced by the length of styrene exposure. If exposure was longer than 4 hours the maximum of excretion was at the end of the exposure time; after short-term exposures (4 h or less) it was somewhat delayed. Maximum excretion of phenylglyoxylic acid was delayed both after short-term and 8-hour exposures. Excretion of the metabolites was diphasic (biexponential). The effective half-lives were found to be independent of the level of exposure. The apparent half-lives (determined in the post-exposure time of 0-16 hours) tended to become prolonged at daily repeated exposures. The ratio of mandelic to phenylglyoxylic acid changed considerably with the level of exposure. In biological monitoring it is advisable to determine both metabolites.     

Antimicrobial profile of ceftazidime: the situation in three larger hospitals in Prague between 1981 and 1983

As a part of preclinical trials of 2,996 clinical isolates from three larger hospitals in Prague were qualitatively and quantitatively assayed for in vitro sensitivity to ceftazidime. In gramnegative bacteria the incidence of resistance to ceftazidime in strain of Enterobacter, Serratia Proteus and Pseudomonas species ranged from 4% to 6% of strains. In grampositive bacteria only strains of enterococci, listeriae and anaerobic bacteria are excluded from the action of this broad-spectrum antibiotic. According to present experiences the antipseudomonal activity of ceftazidime is approximately the same as that of cefsulodine and cefoperazone. Alarmingly, one of the Pseudomonas aeruginosa isolates was found to show a distinct multiresistance to all available lactam, aminoglycoside and broad-spectrum antimicrobials, including ceftazidime.     

Stereochemistry and kinetics of interaction with DNA of the antineoplastic antibiotic olivomycin

The kinetics of interaction of antitumor glycoside antibiotic olivomycin with DNA has been investigated. The existence of two relaxation times in the experimental kinetics curves indicates that two types of antibiotic--DNA complex are formed. We have measured the rate constants of association and dissociation processes and determined their temperature dependences. It is suggested, that one of the complex form results from nonspecific interaction between glycoside residues of the antibiotic molecule and sugar-phosphate backbone of DNA whereas the other type of complex exhibits a pronounced specificity for GC-rich regions on DNA. The binding specificity probably results from formation of a H-bond between the antibiotic chromophore ring and guanine 2-amino group. A stereochemical model for olivomycin-DNA complex is proposed. According to this model the antibiotic chromophore and glycoside residues are located in the narrow groove of DNA.     

Cleavage beyond the block stage and survival after transfer of early bovine embryos cultured with trophoblastic vesicles

Early bovine embryos (1- to 8-cell stages) were recovered from superovulated heifers at slaughter on Days 2 or 3. Embryos were cultured for 3-4 days in Medium B2 supplemented with 15% (v/v) fetal calf serum in the absence (B2SS, 106 embryos) or presence of trophoblastic vesicles (B2SS + TV, 190 embryos). At the end of culture, there were more (P less than 0.001) morulae (greater than or equal to 16 cells) in B2SS X TV (46%) than in B2SS alone (18%) irrespective of the initial cell stage. More 8-cell embryos reached the 16-cell stage than did embryos with less than 8 cells (30% vs 15% in B2SS, P greater than 0.05; 70% vs 41% in B2SS + TV, P less than 0.005). After culture, 102 morulae were transferred non-surgically to temporary recipient heifers (84 embryos cultured in B2SS + TV and 18 in B2SS). After 2 or 3 days, 14 out of 58 embryos from the B2SS + TV group and 3 out of 10 embryos from the B2SS group were recovered as blastocysts. Most blastocysts were deep-frozen and stored for several weeks. After thawing, 10 apparently normal embryos from the B2SS + TV group were transferred non-surgically into 10 recipient heifers. Four pregnancies were induced, but only one embryo survived to term (birth of a normal male calf). It is concluded that trophoblastic vesicles release one or several unknown compound(s) normally present in vivo, promoting the cleavage of early bovine embryos.     

Phytosociological characteristics of theRondeletio correifoliae-Andropogonetum multinervosi ass. nova (Sierra de la Siguanea,Isla de la Juventud,Cuba)

The contribution deals with the phytocenological analysis of theRondeletio correifoliae-Andropogonetum multinervosi, an endemic savanna association occuring on the Siguanea Hills (Sierra de la Siguanea) near the Colony Hotel.     

Foliicole flechten aus zaire (III) Die GattungByssoloma Trevisan

Nine species ofByssoloma Trevisan (Pilocarpaceae) are reported from a collection of foliicolous lichens from Zaire, and includeB. murinum sp. n. In addition,B. usambarense sp. n. is described from adjacent Tanzania. A determination key is provided to all the known African species, and each species is briefly characterized.