Biomedical subjects
Publications and source records attributed to C Segura.
ATPase activity in rat mast cells was studied, assuming that pleural and peritoneal mast cells are different populations. Mast cells were purified with Percoll. Enzymatic activity was found to be 36% higher in pleural cells that in peritoneal cells. Moreover, for both populations results show a Na+-K+ ATPase activity either low or slightly inhibited by ouabain.
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The actions of the polyamines compound 48/80, poly-l-lysine and polymyxin B on rat pleural and peritoneal mast cell secretion have been studied. Unpurified pleural mast cells released more histamine than peritoneal mast cells when stimulated by submaximal concentrations of compound 48/80 and poly-l-lysine, but the same profile of response was observed with polymyxin B in both populations. Dose-response studies of peritoneal and pleural mast cells purified with Percoll and Ficoll and stimulated by polymyxin B showed a decreased sensitivity and decreased maximum response of peritoneal cells when Percoll was used. The maximal response of pleural cells and the sensitivity of peritoneal cells were affected only slightly by Ficoll.
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Information is presented on the plasmid-determined beta-lactamases identified in 204 strains of ampicillin-resistant Enterobacteriaceae. The type most frequently identified was TEM-1 (in 85.3% of the strains), followed by SHV-1 (14.70%). Two types of plasmid-determined beta-lactamase were identified in 20 strains; in 18 of them one of the two was TEM-1 and in 13, SHV-1 (the TEM-1 + SHV-1 combination was observed in 12 strains). In the 41 Klebsiella strains the most frequently identified enzyme was SHV-1 (in 28 of the strains) and the proportion of strains with two plasmid-determined beta-lactamases was higher than in the other species studied.
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The MICs of amoxicillin, mezlocillin and BRL 25,000, a combination of two parts amoxicillin and one part clavulanic acid (2AM + 1CA), were measured for 331 Enterobacteriaceae strains which produced beta-lactamases as demonstrated by nitrocefin. The MIC values for mezlocillin and the combination 2AM + 1CA were very similar for the total number of the strains investigated. When investigated separately according to the bacterial species, three different sensitivity groups were established for the above-mentioned preparations: 1) species with the same or similar sensitivity to mezlocillin and 2AM + 1CA (Escherichia coli and Shigella spp., amoxicillin-resistant strains); 2) species which were more sensitive to mezlocillin than to the combination 2AM + 1CA (Citrobacter spp., Enterobacter cloacae, Serratia spp. and indole-positive Proteus as well as strains of E. coli and Shigella spp. which produce a cephalosporinase and are sensitive to amoxicillin); 3) species which are more sensitive to 2AM + 1CA than to mezlocillin (amoxicillin-resistant Salmonella spp., Proteus mirabilis and Klebsiella pneumoniae). This complementary activity of mezlocillin and 2AM + 1CA against Enterobacteriaceae depended on the beta-lactamases produced.
Cefsulodin is a new second generation cephalosporin with a narrow antibacterial spectrum. Its main use is in the treatment of Pseudomonas aeruginosa infections. It is resistant to the action of the beta-lactamases of gram negative bacilli, especially to that of the cephalosporinases. We have studied 170 strains of P. aeruginosa and 10 of other species of Pseudomonas, determining the MIC by the serial dilution test in solid medium and the sensitivity by the agar diffusion test. With 8 microgram/ml, a concentration which is accepted as a limit of sensitivity from the therapeutic point of view, 90% of the P. aeruginosa strains studied are inhibited; the MIC50 is obtained with 2 microgram/ml. Within the strains considered sensitive (MIC less than 8 microgram/ml) there is a marked dispersion of the zone diameters corresponding to each of the values of MIC. This behavior limits the value of the regression line. We have also studied the MIC of carbenicillin for nearly all the strains and we have also studied the MIC of carbenicillin for nearly all the strains and we have found that 86.58% are sensitive to both preparations, 4.96% are sensitive to cefsulodin and resistant to carbenicillin, 4.02% resistant to cefsulodin and sensitive to carbenicillin and 4.69% are resistant to both preparations. From the clinical point of view the number of strains sensitive (or resistant) is similar with both substances but cefsulodin is in absolute values from 8 to 128 times more day active than carbenicillin. Cefsulodin is administered by a parenteral route and is eliminated by the urine in active form. The recommended therapeutic dose for the treatment of systemic infections is 2 g daily IV. This dose may be increased if it is considered advisable. There is good local tolerance (IM and IV), it is only slightly toxic, hardly alters the intestinal flora and may be used at all ages, in patients with immunological alterations and in patients with an altered renal function.
Cephotaxime (HR 756) is a new cephalosporin antibiotic of wide spectrum and extraordinary activity, suitable for parenteral administration. It is highly resistant to the beta-lactamases of Gram negative bacilli, being thus especially active against microorganisms such as indole-positive Proteus, Serratia marcencens, and others usually resistant to most beta-lactamic antibiotics because of their ability to produce beta-lactamases of high activity. The sensitivity of 355 different bacterial strains to cephotaxime has been determined by the disc method, along with the minimal inhibitory concentration (MIC) of the drug to the same strains, and regression lines have been constructed. The extraordinary activity of cephotaxime against Enterobacteriaceae, Vibrio Cholerae, Haemophilus, and Neisseria meningitidis is noteworthy. Of the 220 strains of Enterobacteriaceae studied, 55% produced beta-lactamase. The MIC of cephotaxime for beta-lactamase forming strains was similar to that for non-forming strains.
A study of the minimal inhibitory concentration (MIC) for 200 strains of Salmonella sp. has been performed with the following beta-lactamic antibiotics: mezlocillin and azlocillin (penicillins), cephaclor, cephamandole, cephuroxime and cephotaxime (cephalosporins), and cephoxytine (cephamycin). The MIC has been compared with that of chloramphenicol, ampicillin, amoxicillin and cotrimoxazole, all of them widely used antibiotics for the treatment of Salmonella infections in Spain. The different species and serotypes of Salmonella studied were all sensitive to all the beta-lactamic antibiotics tested. Of particular relevance is the fact that cephotaxime (HR 756) MIC was extraordinarily low even for strains resistent to the penicillins. The MIC of cephaclor, and oral cephalosporin, was similar to that of the parenteral cephalosporins.
We studied the in vitro activity of ciprofloxacin against 570 strains of ampicillin-resistant Enterobacteriaceae and 286 Pseudomonas aeruginosa strains. 95.26% of the Enterobacteriaceae and 53.45% of the P. aeruginosa were inhibited by 0.1 mg/l of ciprofloxacin. 2 mg/l of ciprofloxacin inhibited all of the Enterobacteriaceae strains and 4 mg/l all of the P. aeruginosa. We compared the activity of ciprofloxacin with that of temocillin in the Enterobacteriaceae strains. In the P. aeruginosa strains, classified according to their susceptibility to carbenicillin and gentamicin, we compared the activity of ciprofloxacin with that of ceftazidime. In the strains studied, the in vitro activity of ciprofloxacin is superior to that of temocillin against the Enterobacteriaceae and to that of ceftazidime against the P. aeruginosa strains.
Bacillus thuringiensis produces d-endotoxins that require proteolytic processing to become active. The activation of the B. thuringiensis subsp. medellin 28 kDa (Cyt1Ab1) cytolytic toxin by trypsin, chymotrypsin and gut extract from Culex quinquefasciatus larvae was analyzed. The Cyt1Ab1 toxin of B. thuringiensis subsp. medellin was processed by all proteases tested to fragments between 23 and 25 kDa, while processing of the Cyt1Aa1 toxin produce fragments between 22.5 and 24.5 kDa. The Cyt1Ab1 toxin was preferentially processed at the alkaline pH of 12. The in vitro proteolytic processing of the Cyt1Ab1 toxin by C. quinquefasciatus larvae midgut extract showed a 25 kDa fragment; a similar result was observed when the activation was performed in the in vivo experiments. The solubilized Cyt1Ab1 toxin and the protease resistant cores generated by in vitro processing showed hemolytic activity but not mosquitocidal activity. Amino terminal sequence of the C. quinquefasciatus gut extract resistant fragment indicated that the cutting site was located between Lys31 and Asp32, with a sequence DDPNEKNNHNS; while for the trypsin-resistant fragment the cutting site was determined between Leu29 and Arg30, and for the chymotrypsin-resistant fragment between Arg30 and Lys31.
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In order to show up the hyperproduction of chromosomic beta-lactamases in strains of Enterobacteriaceae and Pseudomonas aeruginosa we have tested a variant of a technique proposed by Medeiros et al. It is a qualitative technique and, the modification introduced allows errors of interpretation to be avoided, when the strain is a producer of plasmidic beta-lactamase. It is based on evaluating the amount of beta-lactamase in a culture, measured in time taken for the hydrolysis of nitrocefin, with or without the addition of clavulanic acid. We present the results obtained in 526 selected strains: 271 E. coli, 116 E. cloacae and 139 P. aeruginosa. One hundred and twenty for strains hydrolyzed the nitrocefin in the absence of clavulanic acid within 60 seconds. Only 52 (41.94%) of these strains did it when clavulanic acid was present; all of them have been qualified as hyperproducers according to susceptibility to beta-lactam antibiotics and the study and identification of the beta-lactamases by analytic isoelectrofocusing. The hydrolysis was evident before the 15 seconds in the 80% of hyperproducer strains. No false positive results were observed.