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Biomedical subjects

C Padilla

Publications and source records attributed to C Padilla.

At least 19 recordsLinked to original sources

In vitro antibacterial activity of the peptide PsVP-10 against Streptococcus mutans and Streptococcus sobrinus with and without glycocalyx.

The antibacterial activity of the peptide PsVP-10 obtained from Pseudomonas sp. R10 against Streptococcus mutans and Streptococcus sobrinus was investigated. One hundred and twenty strains of S. mutans and 120 strains of S. sobrinus with and without glycocalyx were isolated from saliva samples in trypticase-yeast-cysteine-sucrose-bacitracin (TYCSB) agar. Bacterial identification was made by polymerase chain reaction. Glycocalyx production was observed in modified TYCSB agar and confirmed with a modified version of the microplate adherence assay. The minimum inhibitory concentration (MIC) of PsVP-10 bacteriocin was determined by means of the agar dilution method, and the time of bacterial death was calculated by means of colony-forming unit counts. The MIC of the bacteriocin PsVP-10 for both bacterial species with and without glycocalyx was < 2 mg/L and the time of bacterial death was less than 240 s for all the studied bacterial strains. Thus, bacteriocin PsVP-10 could be an interesting possibility to combat these cariogenic bacterial species.

Anti-Bacterial Agents↗

Plasmid-mediated bacteriocin production by Shigella flexneri isolated from dysenteric diarrhoea and their transformation into Escherichia coli.

AIMS: To determine the production of bacteriocin by Shigella flexneri strains, to relate their production to the presence of dysenteric diarrhoea and to asses the genetic determination of the bacteriocin. METHODS AND RESULTS: One hundred and sixteen strains of Sh. flexneri were isolated from patients with diarrhoea and 49 of them produced bacteriocin active against several Escherichia coli and abacteriocinogenic Sh. flexneri strains. The extrachromosomal DNA isolated from bacteriocinogenic Sh. flexneri strains were used as a substrate to transform E. coli HB-101 cells by means of electroporation. CONCLUSIONS: Only the Sh. flexneri strains isolated from dysenteric diarrhoea produced bacteriocin. It was demonstrated that a plasmid of approx. 3 kb was responsible for the genetic determination of these anti-bacterial substances. SIGNIFICANCE AND IMPACT OF THE STUDY: A 3-kb plasmid that harboured information for the production of bacteriocin by Sh. flexneri strains was described. The production of this bacteriocin may be related to dysenteric diarrhoea produced by these bacterial strains.

Adolescent↗

Periodontal pathogens in atheromatous plaques isolated from patients with chronic periodontitis.

BACKGROUND AND OBJECTIVE: It has been suggested that chronic infections may predispose to cardiovascular disease. The relationship between periodontal disease and cardiovascular disease has been a subject of increasing research in recent years. The isolation and identification of periodontal bacteria from atheromatous plaque can contribute to our knowledge of this vascular disease. The aim of this study was to isolate and identify periodontal bacteria from the periodontal pockets of different patients and to compare them with the microorganisms detected in the atheromatous plaques obtained from the same patients. MATERIAL AND METHODS: Clinical isolates were obtained from 12 patients with periodontal wounds and atheromathous plaques. These samples were cultured in the appropriate bacteriological culture media and incubated in an anaerobic system. Periodontal bacteria were identified using polymerase chain reaction (PCR) assays. RESULTS: From the 12 patients studied, nine presented different periodontopathic bacterial species. In two, Actinobacillus actinomycetemcomitans was present in the periodontal pockets and the respective atheromatous plaques. CONCLUSION: The presence of A. actinomycetemcomitans in atheromatous plaques and the periodontal pockets of the same patients could indicate a role for periodontal pathogenic bacteria in the atherosclerosis disease process.

Aged↗

NeuroD1 in the endocrine pancreas: localization and dual function as an activator and repressor.

The basic helix-loop-helix transcription factor NeuroD1 regulates cell fate in the nervous system but previously has not been considered to function similarly in the endocrine pancreas due to its reported expression in all islet cell types in the newborn mouse. Because we found that NeuroD1 potently represses somatostatin expression in vitro, its pattern of expression was examined in both strains of mice in which lacZ has been introduced into the NeuroD1 locus by homologous recombination. Analysis of adult transgenic mice revealed that NeuroD1 is predominantly expressed in beta-cells and either absent or expressed below the limit of lacZ detection in mature alpha-, delta-, or PP cells. Consistent with a previous report, NeuroD1 colocalizes with glucagon as well as insulin in immature islets of the newborn mouse. However, no colocalization of NeuroD1with somatostatin was detected in the newborn. In vitro, ectopic expression of NeuroD1 in TRM-6/PDX-1, a human pancreatic delta-cell line, resulted in potent repression of somatostatin concomitant with induction of the beta-cell hormones insulin and islet amyloid polypeptide. Additionally, NeuroD1 induced expression of Nkx2.2, a transcription factor expressed in beta- but not delta-cells. Transfection studies using insulin and somatostatin promoters confirm the ability of NeuroD1 to act as both a transcriptional repressor and activator in the same cell, suggesting a more complex role for NeuroD1 in the establishment and/or maintenance of mature endocrine cells than has been recognized previously.

Aging↗

In vitro antibacterial activity of the peptide PsVP-10 against antimicrobial-resistant Enterococcus faecalis isolated from clinical samples.

OBJECTIVE: To analyse the antimicrobial activity of the peptide PsVP-10 against 67 resistant Enterococcus faecalis strains isolated from clinical samples. METHODS: The qualitative disc diffusion method and MIC determinations were used. RESULTS: The presence of several multidrug-resistant phenotypes of E. faecalis was demonstrated, in which there were high MICs to chloramphenicol, tetracycline, vancomycin, cefaloridine, ampicillin and gentamicin. In comparison, the peptide PsVP-10 showed lower MICs against all the multidrug-resistant and susceptible E. faecalis. CONCLUSIONS: There is an urgent need for the development of novel antimicrobial agents against the highly resistant E. faecalis. The present study shows that the peptide PsVP-10 might make a contribution to the solution of this serious problem.

Anti-Bacterial Agents↗

Genes coding structural proteins in the Leishmania braziliensis complex.

Acidic ribosomal P1 and P2b proteins, referred to as P proteins, and histone H3 are reported for first time in the Leishmania braziliensis complex. Deoxyribonucleic acid analysis and multiple sequence alignment suggest that both P proteins may maintain their structural function in the ribosomal stalk, in spite of the high rate of mutations detected. The deduced amino acid sequence of protein P1 showed 51% identity with Trypanosoma cruzi protein P1 and protein P2b showed 61% identity with T. cruzi protein P2b. Another conserved protein, L. (Viannia) braziliensis histone H3, showed 82% and 70% identity with histone H3 of L. (Leishmania) infantum and T. cruzi, respectively. The N-terminal end of this histone is divergent in comparison with the consensus eukaryotic sequence. Their predicted tridimensional structure was designed.

Amino Acid Sequence↗

Production of antimicrobial substances, by hospital bacteria, active against other micro-organisms.

Fifty-nine clinical strains of bacteria, isolated from patients in the Regional Hospital of Talca, were studied. Seventy-four percent of these strains produced antibacterial substances, in comparison with 18% of the same bacterial species obtained from patients from a non-hospital habitat. Almost all the bacteria isolated from hospitalized patients demonstrated in vitro resistance to different antimicrobial agents. Pseudomonas aeruginosa was the most frequent species producing antibacterial substances and its products were of high potency, with a wide spectrum of antimicrobial activity. Cure of plasmid DNA, in most of the antibacterial-producer strains, resulted in the loss of their lethal activity and they also became susceptible in vitro to anti-microbials. These results indicated that such properties are encoded in extrachromosomal DNA. We believe that the knowledge of the antimicrobial activity and resistance to antimicrobials of bacteria from a hospital habitat can help explain the selection and persistence of such strains in this particular ecological niche.

Anti-Bacterial Agents↗

Modulation of human polymorphonuclear leukocyte adherence by cyanopeptide toxins.

The oligopeptides microcystins and nodularins are the most common and abundant cyanotoxins present in diverse water systems. They cause different illnesses in animal and humans, sometimes leading to death, and are responsible for severe environmental problems. Here we demonstrate that both microcystin-LR and N. spumigena nodularin (Nod) significantly enhance the early spontaneous adherence of peripheral polymorphonuclear leukocytes (PMNs) over the concentration range 10(-11)-10(-9) M. However, neither of them affect significantly the late spontaneous adherence or the early or late PMN-stimulated adherence (when cells are treated with formyl-methionyl-leucyl-phenylalanine). Since PMN adherence is a key step in the immune response, our data clearly indicate for the first time the immunomodulatory capacity of cyanopeptide toxins. The low concentrations at which the adherence modulation occurs are similar to the physiological concentrations for natural mammalian peptide hormones. Such concentrations are well below those recommended by other authors and World Health Organization in terms of risk assessment as safe for drinking water (8x10(-10) to 10(-9) M).

Adult↗

Short regimen of praziquantel in the treatment of single brain enhancing lesions.

Twenty-six patients with single enhancing brain lesion (SEL) were openly assigned to receive single-day praziquantel therapy (n=14), or not (n=12). From 14 treated patients, complete resolution was found in 11, partial resolution in two, and the remaining case was later diagnosed as an arteriovenous malformation. Side effects presented in only one patient and remitted in the same day with symptomatic treatment. Conversely, the lesions persisted unchanged in six of 12 patients in the non-treatment group. Untreated patients with persisting lesions were prescribed praziquantel treatment. After this, SELs disappeared in three cases, other diagnoses (brain tuberculoma and arteriovenous malformation) were made in two, and one was not evaluated. When analyzed in regard to the baseline serology, resolution of lesions on computed tomography was found in 13 (complete=12, partial=1) of 14 seropositive patients, whereas it only happened in six (complete=5, partial=1) of 12 seronegative patients. Serological screening defines a subset of SEL patients with good prognosis. If antiparasitic therapy is to be used in patients with SEL, and we cannot find a strong argument against it, single-day praziquantel is the regimen of choice based on duration, costs, and minimal side effects.

Adolescent↗

Molecular basis of glucose-6-phosphate dehydrogenase deficiency among Filipinos.

BACKGROUND: Multiplex polymerase chain reaction (PCR) using multiple tandem forward primers and a common reverse primer (MPTP) was recently established as a comprehensive scanning system for mutations in X-linked recessive diseases. In this report, MPTP was tested to scan for mutations of the glucose-6-phosphate dehydrogenase (G6PD) gene. METHODS: Mutations in exon 11 of the G6PD gene were screened by MPTP in five unrelated Filipino cases with G6PD deficiency. RESULTS: Of the five patients, four screened positive for a mutation in the gene. Sequencing of the amplified products confirmed that three cases had a C-->T substitution at nucleotide (n.t.) 1360 (C1360T) resulting in an amino acid change of arginine to cysteine at position 454 and one had a silent single base substitution C-->T at nucleotide 1311. CONCLUSION: Our results document a C1360T mutation of the G6PD gene in three Filipino patients in the Philippines.

Amino Acid Sequence↗

An outbreak of acute bartonellosis (Oroya fever) in the Urubamba region of Peru, 1998.

During May 1998, we conducted a case-control study of 357 participants from 60 households during an outbreak of acute bartonellosis in the Urubamba Valley, Peru, a region not previously considered endemic for this disease. Blood and insect specimens were collected and environmental assessments were done. Case-patients (n = 22) were defined by fever, anemia, and intra-erythrocytic coccobacilli seen in thin smears. Most case-patients were children (median age = 6.5 years). Case-patients more frequently reported sand fly bites than individuals of neighboring households (odds ratio [OR] = 5.8, 95% confidence interval [CI] = 1.2-39.2), or members from randomly selected households > or = 5 km away (OR = 8.5, 95% CI = 1.7-57.9). Bartonella bacilliformis isolated from blood was confirmed by nucleotide sequencing (citrate synthase [g/tA], 338 basepairs). Using bacterial isolation (n = 141) as the standard, sensitivity, specificity, and positive predictive value of thin smears were 36%, 96%, and 44%, respectively. Patients with clinical syndromes compatible with bartonellosis should be treated with appropriate antibiotics regardless of thin-smear results.

Adolescent↗

Note: purification and characterization of the bacteriocin PsVP-10 produced by Pseudomonas sp.

A bacteriocin (bacteriocin PsVP-10) produced by Pseudomonas sp. R-10 was purified by a simple method that included an extraction of the bacteriocin with chloroform, followed by cation exchange chromatography. The purity of the bacteriocin was verified by RP-HPLC. It is a peptide of 2.4 kDa, very stable to heat, to proteolytic enzymes and to pH. It presents a very broad spectrum of antimicrobial activity against Gram-positive and Gram-negative bacteria.

Amino Acids↗

The expression of the bstVIM gene from Bacillus stearothermophilus V is restricted to vegetative cell growth.

The activity of BstVI DNA methyltransferase was monitored during the sporulative cycle of Bacillus stearothermophilus V. Significant methylase activity was found only in bacteria growing vegetatively. This was confirmed by Northern hybridization, which indicated that the bstVIM gene was not transcribed in cells undergoing sporulation. Supporting evidence came from experiments which demonstrated that the RNA polymerase holoenzyme from these cells did not recognize the promoter elements upstream of the bstVIM gene.

Bacterial Proteins↗

Isolation and characterization of AAP1. A gene encoding an alanine/arginine aminopeptidase in yeast.

The yeast AAP1 gene, encoding a putative amino-peptidase, was isolated based on its ability to suppress the temperature-sensitive growth on nonfermentable carbon sources of spr5, a stationary phase regulatory mutant. AAP1 was physically mapped to chromosome VIII between PUT2 and CUP1. Sequence analysis of the AAP1 gene showed a 1581-nucleotide open reading frame capable of encoding a 59-kilodalton protein. The protein encoded by this open reading frame exhibits approximately 40% sequence identity to human, rat, and mouse aminopeptidases. In limited regions, sequence identity between Aap1 and the mammalian aminopeptidases ranges from 53% to 93%. Insertional inactivation of the AAP1 gene resulted in a decrease in glycogen accumulation and the loss of the major band of arginine/alanine aminopeptidase activity. Strains carrying the AAP1 gene on a high copy plasmid show an increase in the major arginine/alanine aminopeptidase activity, a dramatic increase in glycogen accumulation, and an increase in transcription from a vector carrying lacZ fused to the promoter of a gene (SSA3) expressed during post-diauxic and stationary phases of the culture cycle. We conclude that although the AAP1 gene is not essential for viability, the Aap1 protein positively affects glycogen accumulation in yeast.

Amino Acid Sequence↗

Range of action and genetic bacteriocin codification of Pseudomonas aeruginosa isolated from three different ecological niches.

Strains of Pseudomonas aeruginosa isolated from sediments in wells showed a greater bacteriocinogenic activity in those isolated from river and clinical specimens. More than half of all the strains examined had extrachromosomal DNA. Plasmid DNA was extracted from all the strains and in only 24/120, all from different origins, was the curing achieved; all these strains coded their bacteriocins in the chromosomal DNA.

Bacteriocins↗

Lumbar platyspondyly--characteristic sign of Ehlers-Danlos syndrome.

Three patients with Ehlers-Danlos syndrome have been observed to have isolated flattening of the lumbar vertebral bodies. Flattening of the lumbar vertebrae with normal appearance of the rest of the spine may be an important radiological clue to early recognition or confirmation of the diagnosis of Ehlers-Danlos syndrome.

Adolescent↗

Inhibition of the growth of enteropathogenic bacilli by bacteriocins produced by micro-organisms from the sediment of wells.

The bacterial flora of the sediment of 20 wells of water for human consumption in the rural area of the VII Region in Chile was examined. Fourteen strains of bacteria, from different wells, produced bacteriocins which inhibited the growth of Salmonella typhi, Salm. typhimurium, Shigella sonnei and enterotoxigenic Escherichia coli. About 50% of these strains contained plasmids of different molecular weight and a large number of these codified for bacteriocins. The results suggest what is required to implement an efficient, simple and economical biological system for the purification or control of the number of enteropathogenic bacilli of well water in the rural area.

Bacteriocins↗

[Uropathogenic Serratia marcescens of hospital origin: analysis of the adhesion mechanism of 2 strains].

We isolated 2 Serratia marcescens strains, fim (0) and fim (-), from patients with urinary tract infection. Adhesion properties were studied by hemagglutination, response of uroepithelial cells and electron microscopy techniques. S marcescens fim (+) showed high adherence to uroepithelial cells, hemagglutination capacity and peculiar electron microscopy characteristics. All of these are absent in the fim (-) variety. The fim (+) strain was resistant to nitrofurantoin, nalidixic acid, tetracyclin, ampicillin, gentamycin, cefradine, cephotaxime and cotrimoxazole, whereas the fim (-) strain was sensitive to all. An absence of extrachromosomal DNA was demonstrated in S marcescens fim (+) suggesting that its characteristics are not coded in plasmids.

Bacterial Adhesion↗