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Biomedical subjects

M Blanco

Publications and source records attributed to M Blanco.

At least 199 records · Page 11Linked to original sources
PubMed1991

Enterotoxigenic Escherichia coli associated with infant diarrhoea in Galicia, north-western Spain.

To assess the role of enterotoxigenic Escherichia coli (ETEC) in infantile diarrhoea, 482 children with diarrhoea and 103 healthy controls, from three localities of Galicia, north-western Spain, were investigated between 1985 and 1988. Rotavirus (37.3%) and Salmonella spp. (12.8%) were the most common causal agents, followed by ETEC (3.9%), Campylobacter jejuni (2.3%), Shigella spp. (0.9%) and Yersinia enterocolitica (0.5%). ETEC were significantly more frequently isolated from children with diarrhoea who were under 1 month of age (26.5%) than from older diarrhoeic children (2.2%) (p less than 0.001) or from healthy children who were under 1 month of age (0%) (p less than 0.05). Among children who harboured ETEC, five of the nine children under 1 month of age developed diarrhoea in hospital, whereas none of the 10 children over 1 month of age did so. Seventeen ETEC isolates produced heat-stable enterotoxin (STa) only, four produced only heat-labile enterotoxin (LT), and two produced both toxins. Colonisation factor antigens CFA/I and CFA/II were detected in 11 (55.0%) of the 20 ETEC isolates that remained enterotoxigenic after maintenance in the laboratory. Most ETEC isolates belonged to serotypes O153:K-:H45 (nine STa+ CFA/I+ isolates), O27:K-:H7 (three STa+ isolates) or O6:K15:H16 (two LT+ STa+ CFA/II+ isolates). Our results suggest that ETEC constitute an important cause of neonatal diarrhoea in this part of Spain.

Antigens, Bacterial↗
PubMed1991

Effects of ciprofloxacin on plasmid DNA supercoiling of Escherichia coli topoisomerase I and gyrase mutants.

Changes in plasmid DNA supercoiling were measured following treatment of Escherichia coli cells, carrying topoisomerase mutations, with the quinolone ciprofloxacin. In quinolone-susceptible cells (top+ gyr+) as well as in topA mutants and in gyrB mutants, plasmid DNA was relaxed after the addition of ciprofloxacin. In cells partially resistant to quinolones, low ciprofloxacin levels led to an increase in negative superhelicity of plasmid DNA, whereas at higher ciprofloxacin concentrations, DNA became relaxed. Cells exhibiting partial resistance to quinolones carried either a gyrA mutation alone or a combination of gyrA and gyrB mutations. Moreover, they showed a reduction in gyrase activity, indicated by the supercoiling of a reporter plasmid. Therefore, we conclude that a low level of quinolone action and a DNA with a lower-than-normal level of superhelicity are the two essential conditions for obtaining a ciprofloxacin-promoted increase in plasmid DNA supercoiling. In contrast, deficiency in topoisomerase I is not required for this effect.

Ciprofloxacin↗
PubMed1991

Enterotoxins, colonization factors and serotypes of enterotoxigenic Escherichia coli from humans and animals.

Enterotoxigenic Escherichia coli (ETEC) strains may synthesize both thermolabile (LT-I and LT-II) and thermostable (STa and STb) enterotoxins. Whereas thermolabile enterotoxins are high molecular weight proteins (85,000 d-90,000 d) composed by a single enzymatic A subunit combined with five B subunits which enable toxin for the receptor recognition, thermostable enterotoxins are small peptide chains with molecular weight between 1,900 d and 5,000 d. In addition to the synthesis of enterotoxins, the ability of ETEC strains to cause diarrhoea is also conditioned by the possession of colonization factors which enable bacteria adhere-to and colonize the luminal surface of small bowel. Colonization factors in ETEC strains were located in rigid fimbriae and flexible fibrils constituted by protein subunits ranging in size from 14,500 d to 31,000 d and usually responsible for mannose-resistant haemagglutination with determined erythrocyte species. Both enterotoxins and colonization factors are controlled by plasmids. There exist plasmids which may code separately enterotoxins and colonization factors, and besides there also exist recombinant plasmids coding together these two virulence factors. Human ETEC strains may synthesize LT-I and/or STa enterotoxins, they may possess the colonization factors named CFA/I, CFA/II, CFA/III or CFA/IV, and they belong mainly to serogroups O6, O8, O15, O20, O25, O27, O63, O77, O78, O114, O115, O126, O128, O139, O148, O153, O159 and O167. ETEC strains from porcine origin synthesize LT-I, STa and/or STb, they possess the colonization factors K88, P987, K99 or F41, and they usually belong to serogroups O8, O9, O20, O45, O64, O101, O115, O138, O141, O147, O149 and O157. Bovine and ovine ETEC strains are usually STa producers harbouring on the bacterial surface K99 or F41 colonization factors and they belong to serogroups O8, O9 and O101. Nevertheless, some particular bovine ETEC strains synthesizing LT-II have been described. Thus, a high specificity level between ETEC strains causing diarrhoea in humans and domestic animals can be observed. This is mainly due to the specific recognition between bacterial colonization factors and the epithelial receptors during host-parasite interaction.

Adhesins, Escherichia coli↗
PubMed1990

Comparison of reactogenicity and antigenicity of M37 rotavirus vaccine and rhesus-rotavirus-based quadrivalent vaccine.

90 Venezuelan infants aged 10-20 weeks were randomly allocated to four groups which received one of the following: the M37 vaccine (1 x 10(4) pfu [plaque-forming units]); quadrivalent rotavirus vaccine (1 x 10(4) pfu each of serotype 3 rhesus rotavirus [RRV] and human rotavirus-RRV reassortants of serotypes 1, 2, and 4); balanced quadrivalent vaccine consisting of 1 x 10(4) pfu of serotype 1 and 3 components but 5 x 10(4) pfu of serotype 2 and 4 components; or placebo. The frequencies of transient febrile responses in these four groups were 20%, 27%, 30%, and 9%. 50% of 22 infants tested who received M37 vaccine showed a serum rotavirus IgA antibody response, compared with 74% of the 23 quadrivalent and 86% of the 22 balanced-quadrivalent recipients. 64% of the M37 recipients showed a neutralising antibody response to M37; 27% showed such responses to human serotype 1 Wa strain and 27% to serotype 4 neonatal strain ST3. 17-39% of the quadrivalent recipients and 27-41% of the balanced-quadrivalent recipients showed neutralising antibody responses to serotypes 1-4. 70-73% of the quadrivalent and balanced quadrivalent groups also showed neutralising antibody responses to RRV.

Antibodies, Viral↗
PubMed1990

Comparative evaluation of three tests for the detection of Escherichia coli cytotoxic necrotizing factors (CNF1 and CNF2) using filtrates of cultures treated with mitomycin C.

Necrotizing Escherichia coli (NTEC) strains grown in the presence of mitomycin C released cell associated necrotizing factors CNF1 and CNF2 to culture medium. Using culture filtrates from 96 mitomycin C treated E. coli strains, we have found that a modified HeLa cell assay was a more sensitive and specific method for the detection of CNF1 and CNF2 than the Vero cell assay and the rabbit skin test.

Animals↗
PubMed1990

Revision of the antigenic structure of genus Listeria.

O-antigenic structure of genus Listeria was studied, using antisera (obtained from rabbits) against different O-antigens of reference strains of each serovar. The titres of sera were determined by agglutination using antigens of the same reference strains as well. Some differences from the actual scheme were found: serum antifactor-IX gave a lower titre than expected against antigens 4ab and 6b, while the titre observed against antigen 4b was higher than the expected in this case. Serum antifactor-VIII presented a higher titre than could be expected against antigen 6b. The strains of serovars 4d and 4e used in this experience were impossible to distinguish, and could have been classified in the same serovar. We could not obtain serum antifactor-XI from serovar 6b after several trials. From these differences we propose some modifications of the current antigenic scheme of genus Listeria.

Antibodies, Bacterial↗
PubMed1990

Expression of the recA gene is reduced in Escherichia coli topoisomerase I mutants.

We studied the influence of DNA topological changes on Escherichia coli recA gene expression. This was monitored by measuring beta-galactosidase activity in cells containing a recA-lacZ fusion. To modulate DNA supercoiling we used mutations in the genes encoding for topoisomerase I and DNA gyrase. After either UV irradiation or treatment with the gyrase inhibitor ciprofloxacin, induction of the recA gene was reduced in topA10 mutants, this reduction being alleviated when gyrA or gyrB mutations causing DNA relaxation were present. A reduced induction of recA was also observed after incubation of cells carrying the recA441 mutation at 42 degrees C in the presence of adenine. Using bacteria deficient in the LexA repressor, we have demonstrated that the topA10 mutation reduces the constitutive expression of the recA gene. We suggest that the increase in negative supercoiling resulting from topoisomerase I deficiency interferes with transcription from the recA promoter. The reduction in the expression of the recA gene in topA10 bacteria could determine their increased UV sensitivity as well as their partial defectiveness in SOS mutability.

Ciprofloxacin↗
PubMed1990

Virulence factors of bacteraemic Escherichia coli with particular reference to production of cytotoxic necrotising factor (CNF) by P-fimbriate strains.

Thirty-seven strains of Escherichia coli isolated from bacteraemia and 40 faecal strains isolated from healthy individuals were O serogrouped and investigated for the production of colicins, haemolysin (Hly), cytotoxic necrotising factor (CNF), lethal activity for mice, the expression of P fimbriae, mannose-resistant (MRHA) and mannose-sensitive (MSHA) haemagglutination, and relative cell surface hydrophobicity. Virulence factors significantly associated with bacteraemic strains were: serogroups O2, O4, O6, O7, O8 and O75 (54% versus 10%, p less than 0.001), production of Hly (32% versus 8%, p less than 0.02) and CNF (38% versus 10%, p less than 0.01), expression of P fimbriae (27% versus 5%, p less than 0.02), MRHA types III, IVa and IVb (51% versus 8%, p less than 0.001), and possession of a moderate cell surface hydrophobic charge (35% versus 13%, p less than 0.05). Virulence factors were strongly associated with strains expressing defined MRHA types. Thus, all strains belonging to MRHA types III and IVa were toxigenic, whereas only 11% of strains belonging to MRHA types IVb, V or VI were toxigenic (p less than 0.001). Virulence factors were concentrated in strains belonging to O serogroups usually found in E. coli that cause extra-intestinal infections, especially in strains of O4 and O6 groups. The most interesting result of this study was that all 12 P-fimbriate strains expressed the MRHA type IVa and 11 of them synthesised CNF.

Adolescent↗
PubMed1990

Clinical studies of a quadrivalent rotavirus vaccine in Venezuelan infants.

Phase I studies of an oral quadrivalent rotavirus vaccine were conducted in 130 Venezuelan infants 10 to 20 weeks of age. The vaccine consists of a mixture of equal amounts of rhesus rotavirus (RRV) vaccine (serotype 3 [VP7]) and each of three human rotavirus-RRV reassortant strains: D x RRV (serotype 1 [VP7]), DS1 x RRV (serotype 2 [VP7]), and ST3 x RRV (serotype 4 [VP7]). Three different doses of the quadrivalent vaccine (0.25 x 10(4), 0.5 x 10(4), and 10(4) PFU of each component) were evaluated sequentially for safety and antigenicity in placebo-controlled, double-blind trials. Starting the day after vaccination, the infants were monitored by daily home visits for 7 days. Only minor reactions were observed during this period; these were limited to mild transient febrile episodes which began day 2 or 3 after vaccination and lasted 1 to 2 days in 15 to 30% of the infants. Serological studies demonstrated that 68 to 96% of the infants developed a rotavirus serum immunoglobulin A response following vaccination. However, when tested by plaque reduction neutralization assay against individual human rotavirus serotype 1, 2, 3, or 4, the response rates ranged from 4 to 23% with the low dose, 21 to 33% with the medium dose, and 32 to 58% with the high dose. Most (73 to 79%) infants developed neutralizing antibodies to RRV following administration of each dose schedule. Vaccine virus shedding was analyzed by utilizing tissue culture isolation of virus from stool. All of the infants who received the lower of medium dose and 89% of those fed the high dose shed one or more components of the vaccine. Analyses of rotavirus serotypes isolated from the stool of infants who received the 0.25 x 10(4) -PFU dose revealed that DS1 x RRV was the most commonly shed vaccine component, followed by RRV, D x RRV, and ST3 x RRV in that order.

Administration, Oral↗
PubMed1990

Evidence for two types of cytotoxic necrotizing factor in human and animal clinical isolates of Escherichia coli.

We have characterized the in vitro and in vivo toxic properties of cell sonic extracts from 22 animal and human clinical isolates of Escherichia coli that caused both necrosis in the rabbit skin and multinucleation in tissue cultures, two toxic properties previously reported as being specific for E. coli cytotoxic necrotizing factor (CNF). Two distinct toxic phenotypes were observed. Type 1, which was displayed by originally described CNF strains, was characterized by extensive multinucleation and rounding of cells in HeLa cell culture assays, moderate necrosis in the rabbit skin test, and absence of necrosis in the mouse footpad test. Type 2, which has recently been shown to be associated with E. coli Vir plasmid, was characterized by moderate multinucleation, by polymorphism and elongation of HeLa cells, and by an intense necrotic response in both the rabbit skin test and the mouse footpad test. The distinction between the two cytotoxins accounting for these effects (CNF 1 and CNF 2), together with their partial relatedness, was confirmed by seroneutralization studies of both cytopathic effects and necrosis in the rabbit skin test. In addition, type 2 extracts were more lethal in the mouse intraperitoneal test and induced a moderate, although not totally repetitive, fluid accumulation in the ileal loop test. The original toxic properties of these recently recognized categories of E. coli strains, together with their association with enteritis and septicemia, suggest that these strains may play a significant role in pathology.

Animals↗
PubMed1990

Transcranial magnetic stimulation in epileptic patients: usefulness and safety.

We studied 58 patients with partial or generalized epilepsy who had transcranial magnetic stimulation (TMS) of the brain motor regions. Short-term monitoring disclosed that the stimulation did not provoke seizures or EEG changes in any patient. Long-term follow-up disclosed that the epileptic condition was not made worse by TMS. TMS, as currently used for monitoring conduction in central motor pathways, does not induce seizures in drug-treated epileptic patients.

Adult↗
PubMed1990

Revision of the validity of CAMP tests for Listeria identification. Proposal of an alternative method for the determination of haemolytic activity by Listeria strains.

The validity of CAMP tests with Staphylococcus aureus and Rhodococcus (Corynebacterium) equi as defined for Listeria identification was revised. This characterization method appeared to be unreliable for two reasons: first, a positive CAMP test with R. equi is not specific for Listeria ivanovii as Listeria monocytogenes (and Listeria seeligeri) give also a clear positive reaction; second, doubtful reactions could be observed with S. aureus when assaying haemolytic and non-haemolytic Listeria strains (possibility of false negative and false positive results; subjectivity of the interpretation). The use of a Microplate technique previously described instead of CAMP tests is proposed for the reliable demonstration of the haemolytic character of Listeria in the routine identification of these organisms.

Bacterial Typing Techniques↗
PubMed1990

Exercise-induced anaphylactic reaction to grain flours.

On rare occasions, reproducible exercise-induced anaphylactic reactions (EIA) occur in some patients only after certain foods have been eaten before exercise, yet eating these foods alone or exercising alone causes no symptoms. This special response has been evident sometimes with shellfish, nuts, and wheat. We describe a patient in whom grain flour was a triggering factor for EIA. Skin tests and RAST were positive for grain flours. Normally, the patient tolerated grain flours without symptoms and IgE mechanisms had not been suspected. Testing for food hypersensitivity may be important in patients with EIA.

Anaphylaxis↗
PubMed1989

New Escherichia coli gyrA and gyrB mutations which have a graded effect on DNA supercoiling.

We isolated new gyrA and gyrB mutations in Escherichia coli which have a graded effect on DNA supercoiling. The mutants, selected respectively for resistance to nalidixic acid and coumermycin, were sorted by means of a rapid in vivo assay of DNA gyrase activity (Aleixandre and Blanco 1987). Cells carrying a gyrB (Cour) mutation usually showed a decrease in DNA supercoiling, which would indicate a reduction in gyrase activity. In contrast, most of the gyrA (Nalr) mutations had no significant effect on DNA supercoiling. Moreover, they conferred a high level of resistance to nalidixic acid and other quinolones, thus being similar to the gyrA (Nalr) mutants currently used. We also detected rare gyrA mutants showing a reduction in DNA gyrase activity. These mutants were, in addition, resistant to only low concentrations of quinolones, which allowed us to use the phenotype of partial quinolone resistance as an indicator to score gyrA mutations affecting DNA supercoiling. When gyrB mutations were introduced into the gyrA mutants, these became more sensitive to quinolones and a decrease in supercoiling was observed. Moreover, the topA10 mutation sensitized gyrA (Nalr) cells to quinolones. We conclude therefore that the GyrA-dependent quinolone resistance is diminished as a consequence of the reduction either in topoisomerase I or gyrase activities.

Aminocoumarins↗
PubMed1989

Prevalence of enterotoxigenic Escherichia coli strains in outbreaks and sporadic cases of diarrhoea in Spain.

Escherichia coli strains isolated 1985-1988 in Spain from patients with diarrhoea were examined; 1170 strains were isolated from 582 sporadic cases of diarrhoea in children, and seven strains were associated with seven outbreaks of diarrhoea. Strains positive for STa enterotoxin production in the infant mouse test were also assayed for production of LT enterotoxin on Vero cells and by a coagglutination test. Thirty-one strains were STa positive: 28 were isolated from 16 (2.7%) sporadic cases of diarrhoea and three were responsible for outbreaks. The majority of STa+LT- strains from both outbreaks and sporadic cases were serotype O153:H45 and expressed the CFA/I colonization factor antigen. Enterotoxigenic STa+LT- strains of serotype O27:H7 and STa+LT+ CFA/II+ strains of serotype O6:K15:H16 were also isolated frequently from sporadic cases.

Antigens, Bacterial↗
PubMed1989

Enterotoxic, cytotoxic, necrotic and lethal activities in cell-free extracts of Salmonella strains isolated from humans.

Unconcentrated cell-free sonic extracts from thirty Salmonella strains isolated from the faeces and blood of humans were investigated for the production of enterotoxins in various tests (Vero cell, infant mouse, rabbit skin permeability and rabbit ileal loop), as well as for lethal activity in adult mice. Sonic extracts from 23 (76.7%) strains were lethal for mice, 21 (70%) increased skin permeability and 3 (10%) showed necrotizing activity for the rabbit skin. No Salmonella strain producing typical Escherichia coli toxins, such as thermolabile (LT) or thermostable (STa) enterotoxins, Verotoxin (VT) or cytotoxic necrotizing factor (CNF) cytotoxins, were detected. Non-repetitive fluid accumulation in rabbit loops was obtained when unconcentrated sonic extracts from 10 selected strains were assayed in seven rabbits. Growth of Salmonella in casamino acid yeast extract medium, followed by treatment of bacterial cells with polymyxin B, was demonstrated to be a rapid and sensitive method for releasing the delayed permeability factor.

Animals↗