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Biomedical subjects

M Blanco

Publications and source records attributed to M Blanco.

At least 217 records · Page 12Linked to original sources

Reactions to and antigenicity of two human-rhesus rotavirus reassortant vaccine candidates of serotypes 1 and 2 in Venezuelan infants.

The reactions to and antigenicity of two human-rhesus rotavirus (RRV) reassortants (human rotavirus strain D x RRV and human rotavirus strain DS1 x RRV) with the VP7 neutralization specificity of a serotype 1 or serotype 2 rotavirus were evaluated in a placebo-controlled double-blind trial in 116 1- to 5-month-old infants in Caracas, Venezuela. The children were randomly divided into five groups to receive orally the following inocula: (i) 10(4) PFU of D x RRV reassortant; (ii) 10(4) PFU of DS1 x RRV reassortant; (iii) 10(4) PFU of RRV; (iv) 5 x 10(3) PFU of D x RRV and 5 x 10(3) PFU of RRV; and (v) placebo. The children were examined daily for 7 days following vaccine administration; 8 to 26% of the vaccinated infants developed a mild febrile reaction which in most cases lasted only 1 day. Seroresponses to rotavirus were observed in 39 to 65% of the vaccinees by plaque neutralization assay and in 57 to 88% by an immunoglobulin A enzyme-linked immunosorbent assay. Vaccine shedding was detected in 53 to 86% of the vaccinees. Analysis of neutralization antibody responses indicates that the VP4 protein represents an important component of the response induced by the vaccines.

Age Factors↗

Direct isolation (without enrichment) of Listeria monocytogenes in milk.

In the present work we have developed a direct method (without enrichment) for the isolation of listeriae in milk, using the combinated action of SDS (sodium dodecyl sulphate), centrifugation and selective media. It is possible to identify even 0.5 listeriae/ml amongst 7.2 x 10(7) c.f.u. contaminants/ml.

Animals↗

UV-light-induced mutability in Salmonella strains containing the umuDC or the mucAB operon: evidence for a umuC function.

Multicopy plasmids carrying either the umuDC operon of Escherichia coli or its analog mucAB operon, were introduced into Ames Salmonella strains in order to analyze the influence of UmuDC and MucAB proteins on repair and mutability after UV irradiation. It was found that in uvr+ bacteria, plasmid pICV80:mucAB increased the frequency of UV-induced His+ revertants whereas pSE117:umuDC caused a smaller increase in UV mutagenesis. In delta uvrB bacteria, the protective role of pSE117 against UV killing was weak, and there was a great reduction in the mutant yield. In contrast, in these cells, pICV80 led to a large increase in both cell survival and mutation frequency. These results suggest that in Salmonella, as in E. coli, MucAB proteins mediate UV mutagenesis more efficiently than UmuDC proteins do. Plasmid pICV84:umuD+ C- significantly increased UV mutagenesis of TA2659: delta uvrB cells whereas in them, pICV77:mucA+ B- had no effect on mutability indicating the presence in Salmonella TA2659 of a gene functionally homologous to umuC.

Bacterial Proteins↗

Production of toxins by Escherichia coli strains isolated from calves with diarrhoea in galicia (north-western Spain).

A total of 289 Escherichia coli colonies isolated from 78 diarrhoeic calves were studied for production of heat-labile (LT) and heat-stable (STa) enterotoxins, verotoxin (VT), cytotoxic necrotizing factor (CNF) and K99 antigen, and they were serotyped. Production of STa was detected in a single strain possessing both K99 and F41 antigens; the serotype was 09:K (A) 35. LT-producing strains were not detected. From 16 (20.5%) calves, 51 VT-producing colonies of E. coli were isolated. Production of the necrotic factor was detected in 33 E. coli colonies isolated from 14 (17.9%) calves. Serotype was a useful marker for production of VT and CNF. Among the 51 VT-producing colonies, 24 were untypable and the remainder belonged to serotypes O2:K?, O103:K--, O104:K?, O128:K?, O153:K-- and O157:K--:H7. Four of the 33 CNF-producing colonies were untypable and the majority of the remaining colonies belonged to serotypes O15:K14, O78:(K80), O123:K-- and O139:K--. Both VT and CNF were lethal for mice, but only CNF showed necrotizing reaction in rabbit skin. Our results indicate that VT-producing and CNF-producing E. coli strains are frequently isolated from diarrhoeic calves and that according to the serotypes exhibited, some of them might be considered potential pathogens for humans. The role of VT-producing and CNF-producing strains in calf diarhoea remains to be established.

Animals↗

Simultaneous multiwavelength spectrophotometric quantitation of active components in analgesic formulations. Comparative study of three calculation methods.

This work deals with the UV spectrophotometric quantitation of a mixture of compounds with overlapped spectra. The mixture spectrum is resolved by use of three computational programs based on different algorithms, namely Multicomponent Analysis (commercial software available from Hewlett-Packard), MULTIC (relying on multiple regression analysis) and SIMPLEX. The results obtained for mixtures of acetylsalicylic acid, acetaminophen and caffeine in commercial analgesic formulations, are compared.

Journal Article↗

Toxins and serotypes of faecal non-enterotoxigenic and non-enteropathogenic Escherichia coli strains causing mannose-resistant haemagglutination: relation with haemagglutination patterns.

Forty-three faecal non-enterotoxigenic and non-enteropathogenic human Escherichia coli strains causing mannose-resistant haemagglutination (MRHA) were tested for production of cytotoxic necrotizing factor (CNF), haemolysis (Hly), Verotoxin (VT) and lethal activity for mice. The serotypes of the strains were also determined. Of the total strains investigated, 49% synthesized CNF, 53% were haemolytic and 40% were lethal for mice. No strain producing VT was detected. Striking differences in the production of Hly and CNF were observed when MRHA strains were grouped according to their lethal or non-lethal activity. Thus, 82% of lethal strains produced Hly and/or CNF whereas only 35% (p less than 0.01) and 27% (p less than 0.01) of non-lethal strains produced Hly and CNF, respectively. The production of toxins was specially associated with strains possessing defined MRHA types. Thus, 100%, 82% and 50% of strains belonging to MRHA types III, IVa and V, respectively, were toxigenic, whereas no toxigenic strains from MRHA types IVb and VI were detected. The majority (77%) of MRHA strains possessed typical O groups usually reported to be present in pathogenic extraintestinal E. coli or in facultatively enteropathogenic E. coli. Furthermore, these O groups were more frequently detected in toxigenic (93%) than in non-toxigenic (47%) strains (p less than 0.01). Our results suggest that faecal non-enterotoxigenic E. coli strains belonging to MRHA types III, IVa and V may be responsible for extraintestinal infections as well as for sporadic intestinal infections, and that certain O groups are specially associated with E. coli strains belonging to particular MRHA types.

Animals↗

Extreme somatosensory evoked potentials (ESEPs) elicited by tapping of hands or feet in children: a somatosensory cerebral evoked potentials study.

Six children with tactile-evoked spikes in the EEG, also defined as extreme somatosensory evoked potentials (ESEPs), underwent an SEPs study in order to define the characteristics of such evoked potentials. Short-latency SEPs showed normal mean latency and amplitude values. Mid- or long-latency SEPs of abnormally high amplitude were recorded after stimulation of one or more extremities. Such extreme responses which showed the same reactivity proper to normal long-latency SEPs could be considered to correspond to the evoked spikes in the EEG.

Child↗

Effect of acid inhibition on Campylobacter pylori.

Campylobacter pylori (Cp) infection has been demonstrated in 59 of 65 (91%) patients with gastric or duodenal ulcer. Patients were included in a double blind study with ranitidine or famotidine for 4-8 weeks. After therapy, all gastric and duodenal ulcers healed but Cp remained in all previously infected patients, associated with chronic inflammation of gastric mucosa. Our results suggest that H2 antagonists do not have any effect either on Cp infection or on concurrent chronic gastritis. At the same time we observed that Cp infection does not disturb the ulcer healing process. However, Cp may be implicated in the persistence of the chronic gastritis and relapses of ulcers.

Adult↗

Heterogeneity in the level of ampicillin resistance conferred by pBR322 derivatives with different DNA supercoiling.

Cloning of an EcoRI restriction fragment, containing the 900 bp gamma-terminal sequence of transposon Tn1000, into pBR322, resulted in two plasmids, pICV63 and pICV64, which differed in the orientation of the cloned fragment within the replicon and in the level of ampicillin resistance conferred on the host cell. The DNAs of these plasmids differ in superhelicity and we suggest that a change in supercoiling of pICV63 DNA leads to this plasmid conferring resistance to only low levels of ampicillin, probably by reducing the expression of the bla gene. This hypothesis is supported by the fact that topA or supX mutations, which abolish topoisomerase I, reduce still further the level of resistance to ampicillin of pICV63-containing cells, whereas the gyrB226 compensatory mutation renders these cells more ampicillin resistant. Plasmid pICV63, therefore, enables mutant alleles of genes governing DNA topology to be recognized.

Ampicillin↗

Mutagenic DNA repair in Escherichia coli. XIII. Proofreading exonuclease of DNA polymerase III holoenzyme is not operational during UV mutagenesis.

We have introduced a mutD5 mutation (which results in defective 3'-5'-exonuclease activity of the epsilon proofreading subunit of DNA polymerase III holoenzyme) into excision-defective Escherichia coli strains with varying SOS responses to UV light. MutD5 increased the spontaneous mutation frequency in all strains tested, including recA430, umuC122::Tn5, and umuC36 derivatives. It had no effect on UV mutability or immutability in any strain or on misincorporation revealed by delayed photoreversal in UV-irradiated umuC36, umuC122::Tn5, or recA430 bacteria. It is concluded that the epsilon proofreading subunit of DNA polymerase III holoenzyme is excluded, inhibited, or inoperative during misincorporation and mutagenesis after UV.

Arginine↗

Different efficiency of UmuDC and MucAB proteins in UV light induced mutagenesis in Escherichia coli.

Two multicopy plasmids carrying either the umuDC or the mucAB operon were used to compare the efficiency of UmuDC and MucAB proteins in UV mutagenesis of Escherichia coli K12. It was found that in recA+ uvr+ bacteria, plasmid pIC80, mucAB+ mediated UV mutagenesis more efficiently than did plasmid pSE117, umuDC+. A similar result was obtained in lexA41 (Def) cells, excluding the possibility that this was due to a differential regulation by LexA of the umuDC and mucAB operons. We conclude that some structural characteristic of the UmuDC and MucAB proteins determines their different efficiency in UV mutagenesis. This characteristic could be also responsible for the observation that in the recA430 mutant, pIC80 but no pSE117 can mediate UV mutagenesis. In the recA142 mutant, pIC80 also promoted UV mutagenesis more efficiently than pSE117. In this mutant, the recombination proficiency, the protease activity toward LexA and the mutation frequency were increased by the presence of adenine in the medium. In recA+ uvrB5 bacteria, plasmid pSE117, umuDC caused both an increase in UV sensitivity as well as a reduction in the mutation frequency. These negative effects resulting from the overproduction of UmuDC proteins were higher in recA142 uvrB5 than in recA+ uvrB5 cells. In contrast, overproduction of MucAB protein in excision-deficient bacteria containing pIC80 led to a large increase in the mutation frequency. We suggest that the functional differences between UmuDC and MucAB proteins might be due to their different dependence on the direct role of RecA protease in UV mutagenesis.

Bacterial Proteins↗

New role for photoreversible pyrimidine dimers in induction of prototrophic mutations in excision-deficient Escherichia coli by UV light.

UV mutagenesis to His+ in certain recA441 lexA51 bacteria was not photoreversible, indicating that pyrimidine dimers are not target lesions. Photoreversibility was observed in recA+ lexA51 bacteria, showing that pyrimidine dimers are needed to activate the recA+ protein (unlike the recA441 protein) to perform a function in UV mutagenesis distinct from cleavage of the lexA repressor.

DNA Repair↗

Ectopic Purkinje cells in the cerebellar white matter of normal adult rodents: a Golgi study.

In Golgi/Río-Hortega preparations of rat and rabbit cerebellar vermis we have occasionally found isolated ectopic Purkinje cells in the white matter. They were located beneath the bases of the folia and their dendritic branches extended within the confines of the white matter without penetrating into the overlying cortical layers. The general morphology of these ectopic cells was variable, particularly in the extension and shape of the dendritic trees, but all of them exhibited a lower density of dendritic branches than normal Purkinje cells. The less-developed ectopic neurons had multipolar dendritic trees with nonplanar branches irregularly studded with spines. The well-developed ones displayed a more extensive arborization of their processes and they usually preserved some morphological features of normal cortical Purkinje cells: distal dendritic branches studded with numerous spines, a pear-shaped soma, clearly defined morphological polarity and a tendency to display planar arrangement of the dendritic arbors. In semithin sections these neurons also showed cytological features of normal Purkinje cells, such as the Nissl substance forming a nuclear cap oriented toward the dendritic pole. We suggest that the abnormal location of the neurons results from a disorder of Purkinje cell migration which occurs naturally during the prenatal development of the cerebellum. The possible morphogenetic mechanisms involved in the migration and differentiation of these ectopic neurons are also discussed.

Animals↗

[Campylobacter in gastric pathology].

The incidence of Campylobacter pyloridis in the stomach of patients with chronic gastritis, peptic ulceration and in normal subjects, has been studied. Seventy-one biopsy specimens were taken from affected and normal gastric mucosa of 61 patients and 10 normal subjects, respectively. In 54 (88.5%) out of the 61 patients and in 1 (10%) out of the 10 normal subjects, C. pyloridis was observed. In addition, in 39 samples this microorganism was successfully cultured.

Campylobacter↗

The perineuronal net in the fastigial nucleus of the rat cerebellum. A Golgi and quantitative study.

The morphological study of the rat fastigial nucleus with the Golgi-Rio Hortega method showed the presence of glial perineuronal nets surrounding the large neurons, but not the small ones. This perineuronal net appeared as a mesh of tenuous glial processes which covers the neuronal perikarya and proximal dendrites. The small alveolate compartments in this mesh seem to correspond to the holes for the synaptic boutons. Our results also indicate that the perineuronal net is derived from interneuronal protoplasmic and velate astrocytes. Using camera lucida drawings of this perineuronal net we have made a quantitative estimation of the size and density of synaptic boutons on these large neurons. The average numerical density of synaptic boutons was about 19 per 100 micron 2 of the neuronal surface, the mean area of the synaptic holes being 2.5 micron 2. Furthermore, the quantitative data evidence that about 52.5% of the neuronal surface is presumably occupied by synaptic boutons whereas the remaining 47.5% is covered by the glial processes of the perineuronal net. Semithin sections prepared from thick Golgi sections were used for the cytological study of the neurons surrounded by this glial pericellular network. The possible functional significance of the perineuronal net in the regulation of synaptic transmission in the fastigial cerebellar nucleus is briefly discussed.

Animals↗