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Biomedical subjects

S Merino

Publications and source records attributed to S Merino.

At least 37 records · Page 2Linked to original sources

Determination by fluorimetric titration of the ionization constants of ciprofloxacin in solution and in the presence of liposomes.

A fluorescence titration method was applied for the determination of pKa of ciprofloxacin (CPX) in solution. Values of 6.18 +/- 0.05 and 8.76 +/- 0.03 were obtained for pKa1 and pKa2, respectively. The method was used to determine the ionization constants in the presence of liposomes of dipalmitoylphosphatidylcholine (DPPC) and DPPC with 10 mol% of dipalmitoylphosphatidylglycerol. A dependence on the surface charge of liposomes was found which supported the existence of a basic electrostatic interaction between CPX and the phospholipid bilayer. Both pK values for the N-4 butyl-piperazinyl derivative (BCPX) of the parent compound were also determined in solution and in the presence of liposomes. The competition of both drugs for the same binding site as 1-anilino-8-naphtalene sulfonate demonstrate that the interaction is governed by electrostatic forces.

Anti-Infective Agents↗

Effects of spatial and host variables on hematozoa in white-crowned sparrows wintering in Baja California.

A survey of blood parasites was conducted in February 1995 on white-crowned sparrows (Zonotrichia leucophrys) wintering in two environmentally different localities of Baja California Sur (Mexico). Blood parasite prevalence was higher in La Purísima (49%) than in San José del Cabo (8%), but there were no differences between ages or sexes within each locality. All haematozoa infections were by Haemoproteus coatneyi, except one bird in each site that were positive for Trypanosoma sp. We found no evidence for the predicted negative relationship between host body condition and intensity of parasitism. The relatively high prevalence in one site suggests that an increase of hematozoa transmission may occur in that area.

Animals↗

[Influence of bispectral index monitoring on fentanyl requirements during total intravenous anesthesia for major gynecological surgery].

OBJECTIVE: To determine the influence that bispectral index (BIS) monitoring of hypnosis might have on need for analgesia during surgery under total intravenous anesthesia provided by bolus administration of fentanyl. PATIENTS AND METHOD: Prospective, randomized and partially double-blind study of 40 patients undergoing major gynecological surgery under total intravenous anesthesia with propofol and fentanyl. In the BIS group (n = 20) propofol administration was adjusted to maintain BIS between 40 and 60. In the control group (n = 20) standard doses were given: 10 mg/kg-1/h-1 after anesthetic induction and for 5 minutes, 8 mg/kg-1/h-1 over the next 5 minutes and 6 mg/kg-1/h-1 throughout the rest of the operation. All patients received intravenous bolus administration of 150 or 75 microg of fentanyl to maintain analgesia whenever systolic blood pressure and heart rate increased 20% over baseline. We compared propofol and fentanyl requirements, intraoperative changes in BIS, and awakening from anesthesia. RESULTS: Patient and surgical characteristics were similar in both groups. BIS monitoring allowed propofol administration to be decreased a mean 24% during maintenance of anesthesia, and this in turn was associated with a significant increase in mean dose of fentanyl (415 microg versus 253 microg in the BIS and control groups, respectively; p = 0.01). Mean values of BIS were higher in the BIS group (46.4 versus 42.2; p = 0.04) and patients in the BIS group awoke sooner (in 7.7 min versus 11.1 min; p = 0.01) and tended to report less pain upon arrival at the postanesthetic recovery room, although the difference was not statistically significant. CONCLUSIONS: BIS monitoring of depth of hypnosis can influence requirements for fentanyl during total intravenous anesthesia by bolus dosing for maintenance of analgesia. This is probably due to changes in the administration of propofol made possible by BIS monitoring.

Adult↗

Are avian blood parasites pathogenic in the wild? A medication experiment in blue tits (Parus caeruleus).

The Hamilton and Zuk hypothesis on haemoparasite-mediated sexual selection and certain studies of reproductive costs are based on the assumption that avian blood parasite infections are detrimental to their hosts. However, there is no experimental evidence demonstrating harmful effects of blood parasites on fitness in wild populations, it even having been suggested that they may be non-pathogenic. Only an experimental manipulation of natural blood parasite loads may reveal their harmful effects. In this field experiment we reduced through medication the intensity of infection by Haemoproteus majoris and the prevalence of infection by Leucocytoazoon majoris in blue tits (Parus caeruleus), and demonstrated detrimental effects of natural levels of infection by these common parasite species on host reproductive success and condition. The fact that some of the costs of infection were paid by offspring indicates that blood parasites reduce parental working capacity while feeding nestlings. Medicated females may be able to devote more resources to parental care through being released from the drain imposed upon them by parasites and/or through a reduced allocation to an immune response. Therefore, this work adds support to previous findings relating hosts' life-history traits and haematozoan infections.

Animals↗

Capsular polysaccharide is a major complement resistance factor in lipopolysaccharide O side chain-deficient Klebsiella pneumoniae clinical isolates.

We have previously demonstrated the existence of Klebsiella pneumoniae clinical isolates deficient in the lipopolysaccharide O side chain, the major factor for resistance to complement-mediated killing in this bacterial species. These isolates are complement resistant, and their mechanisms to resist complement were investigated by selecting transposon-generated complement-sensitive mutants. One mutant with a drastically reduced capacity to grow in nonimmune human serum carried the transposon inserted in an open reading frame of a gene cluster involved in capsule synthesis. This mutant produced less capsule, bound more molecules of the complement component C3, and was more sensitive to complement-mediated and opsonophagocytic killings than was the parent strain. Four additional clinical isolates representing four different K serotypes were studied, and results showed that capsular polysaccharide is a major complement resistance factor in these O side chain-deficient isolates.

Bacterial Capsules↗

Cloning, sequencing, and role in serum susceptibility of porin II from mesophilic Aeromonas hydrophila.

We cloned and sequenced the structural gene for Aeromonas hydrophila porin II from strain AH-3 (serogroup O:34). The genetic position of this gene, like that of ompF in Escherichia coli, is adjacent to aspC and transcribed in the same direction. However, upstream of the porin II gene no similarities with E. coli were found. We obtained defined insertion mutants in porin II gene either in A. hydrophila (O:34) or A. veronii sobria (serogroup O:11) serum-resistant or -sensitive strains. Furthermore, we complemented these mutants with a plasmid harboring only the porin II gene, which allowed us to define the role of porin II as an important surface molecule involved in serum susceptibility and C1q binding in these strains.

Aeromonas hydrophila↗

Cloning and sequencing of the Klebsiella pneumoniae O5 wb gene cluster and its role in pathogenesis.

One representative recombinant clone encoding Klebsiella pneumoniae O5-antigen lipopolysaccharide (LPS) was found upon screening for serum resistance in a cosmid-based genomic library of K. pneumoniae KT769 (O5:K57) introduced into Escherichia coli DH5alpha. A total of eight open reading frames (wb(O5) gene cluster) were necessary to produce K. pneumoniae O5-antigen LPS in E. coli K-12. The enzymatic activities proposed for the wb(O5) gene cluster are in agreement with the activities proposed for the biosynthesis of K. pneumoniae O5-antigen LPS. Using the complete DNA sequence of the K. pneumoniae wb(O5) gene cluster, we obtained (by single or double recombination) genetically well-characterized mutants devoid only of this O5-antigen LPS. Finally, using these O5(-) mutants and the corresponding wild-type strains or complemented mutants with the wb(O5) gene cluster (O5(+) strains), we found that the presence of K. pneumoniae O5-antigen LPS is essential for some pathogenic features like serum resistance, adhesion to uroepithelial cells, and colonization (experimental infections) of the urinary tract in rats.

Amino Acid Sequence↗

[Massive hemorrhage due to esophageal varices as a form of presentation of autoimmune hepatitis].

Autoimmune hepatitis is a necro-inflammatory disease of unknown etiology. An acute or fulminant presentation is rare. We report a 18-year-old man presented with acute variceal hemorrhage. The patient was treated with endoscopic sclerotherapy, somatostatin, balloon tamponade and TIPS. The patient died within a week of massive bleeding and hepatic failure. This patient's history strongly suggests that autoimmune hepatitis can be presented as fulminant hemorrhage. The importance of diagnosis and treatment is emphasized to improve prognosis.

Acute Disease↗

Post-Paget telangiectatic osteosarcoma of the skull.

Sarcomatous transformation is the most dreaded complication of Paget's disease. We report on a case of post-Paget telangiectatic osteosarcoma of the skull, a variant of osteogenic osteosarcoma, in a 79-year-old woman. We discuss the radiological pattern in relationship to the differential diagnosis.

Aged↗

Surface antigen exposure by bismuth dimercaprol suppression of Klebsiella pneumoniae capsular polysaccharide.

The bacterial capsule is an important virulence determinant in animal and plant disease. Bacterial capsule and slime can be inhibited by bismuth compounds, especially when complexed with lipophilic thiol chelators. Bismuth dimercaprol (BisBAL) at 1 ppm of Bi3+ repressed Klebsiella pneumoniae capsule expression in defined medium by nearly 90%, which exposed subsurface structures. The phagocytic index for BisBAL-treated bacteria increased from <10 to 360 bacteria per 100 neutrophils in the presence of complement and anticapsular or anti-O antigen antiserum. BisBAL treatment also enhanced the reactivity of monoclonal antibodies (MAbs) specific for the O1-antigen lipopolysaccharide (LPS) or the LPS core in a dose-dependent manner as indicated by the results of enzyme-linked immunosorbent assays. When anti-O1 MAb was used, the reactivity increased significantly for fully encapsulated O1:K1 or O1:K2 cells but not for O1:K- cells. Deposition of C3b also increased significantly for BisBAL-treated O1:K1 or O1:K2 cells but not for O1:K- cells. Survival of a serum-sensitive strain was <0.1% when nonimmune human serum absorbed with O1:K1 cells was used and 107% when BisBAL-treated cells were used for absorption. Outer membrane proteins were also more accessible on the surface of K. pneumoniae after BisBAL treatment. Thus, at subinhibitory levels, BisBAL inhibited capsule expression, which promoted phagocytosis, enhanced the reactivity of specific antibodies for LPS O antigen, LPS core epitopes, or outer-membrane proteins, and enhanced complement interaction with encapsulated K. pneumoniae. By unmasking bacterial surface structures and enhancing the immune system reactivity to bacteria, bismuth thiols may prove useful as adjuncts for vaccination.

Animals↗

Cloning, sequencing, and role in virulence of two phospholipases (A1 and C) from mesophilic Aeromonas sp. serogroup O:34.

Two different representative recombinant clones encoding Aeromonas hydrophila lipases were found upon screening on tributyrin (phospholipase A1) and egg yolk agar (lecithinase-phospholipase C) plates of a cosmid-based genomic library of Aeromonas hydrophila AH-3 (serogroup O34) introduced into Escherichia coli DH5alpha. Subcloning, nucleotide sequencing, and in vitro-coupled transcription-translation experiments showed that the phospholipase A1 (pla) and C (plc) genes code for an 83-kDa putative lipoprotein and a 65-kDa protein, respectively. Defined insertion mutants of A. hydrophila AH-3 defective in either pla or plc genes were defective in phospholipase A1 and C activities, respectively. Lecithinase (phospholipase C) was shown to be cytotoxic but nonhemolytic or poorly hemolytic. A. hydrophila AH-3 plc mutants showed a more than 10-fold increase in their 50% lethal dose on fish and mice, and complementation of the plc single gene on these mutants abolished this effect, suggesting that Plc protein is a virulence factor in the mesophilic Aeromonas sp. serogroup O:34 infection process.

Aeromonas↗

Genetic analysis of the Serratia marcescens N28b O4 antigen gene cluster.

The Serratia marcescens N28b wbbL gene has been shown to complement the rfb-50 mutation of Escherichia coli K-12 derivatives, and a wbbL mutant has been shown to be impaired in O4-antigen biosynthesis (X. Rubirés, F. Saigí, N. Piqué, N. Climent, S. Merino, S. Albertí, J. M. Tomás, and M. Regué, J. Bacteriol. 179:7581-7586, 1997). We analyzed a recombinant cosmid containing the wbbL gene by subcloning and determination of O-antigen production phenotype in E. coli DH5alpha by sodium dodecyl sulfate-polyacrylamide electrophoresis and Western blot experiments with S. marcescens O4 antiserum. The results obtained showed that a recombinant plasmid (pSUB6) containing about 10 kb of DNA insert was enough to induce O4-antigen biosynthesis. The same results were obtained when an E. coli K-12 strain with a deletion of the wb cluster was used, suggesting that the O4 wb cluster is located in pSUB6. No O4 antigen was produced when plasmid pSUB6 was introduced in a wecA mutant E. coli strain, suggesting that O4-antigen production is wecA dependent. Nucleotide sequence determination of the whole insert in plasmid pSUB6 showed seven open reading frames (ORFs). On the basis of protein similarity analysis of the ORF-encoded proteins and analysis of the S. marcescens N28b wbbA insertion mutant and wzm-wzt deletion mutant, we suggest that the O4 wb cluster codes for two dTDP-rhamnose biosynthetic enzymes (RmlDC), a rhamnosyltransferase (WbbL), a two-component ATP-binding-cassette-type export system (Wzm Wzt), and a putative glycosyltransferase (WbbA). A sequence showing DNA homology to insertion element IS4 was found downstream from the last gene in the cluster (wbbA), suggesting that an IS4-like element could have been involved in the acquisition of the O4 wb cluster.

ATP-Binding Cassette Transporters↗

Klebsiella pneumoniae lipopolysaccharide O typing: revision of prototype strains and O-group distribution among clinical isolates from different sources and countries.

We have previously described an inhibition enzyme-linked immunosorbent assay method for the O typing of O1 lipopolysaccharide from Klebsiella pneumoniae which overcomes the technical problems and limitations of the classical O-typing method. In this study, we have extended the method to all of the currently recognized O types. The method was validated by studying the prototype strains that have defined the O groups by the classical tube agglutinatination O-typing method. Based on these results, we confirmed the O types of 60 of 64 typeable strains, and we propose a revised O-antigenic scheme, with minor but necessary changes, consisting of serogroups or serotypes O1, O2, O2ac, O3, O4, O5, O7, O8, and O12. Application of this typing method to 638 K. pneumoniae clinical isolates from Denmark, Spain, and the United States from different sources (blood, urine, and others) showed that up to 80% of these isolates belong to serotypes or serogroups O1, O2, O3, and O5, independently of the source of isolation, and that a major group of nontypeable isolates, representing about 17% of the total, consists of half O+ and half O- strains. Differences were observed, however, in the prevalence of the lipopolysaccharide O types or groups, depending on the country and isolation source.

Denmark↗

CT evaluation of gastric wall pathology.

The purpose of this study is to show the CT features of common and infrequent pathological lesions of the gastric wall. Although CT features are not often specific, familiarity with the most frequent pathological gastric findings on CT can assist in differential diagnosis.

Diagnosis, Differential↗

Transthoracic needle aspiration biopsy: value in the diagnosis of mycobacterial lung opacities.

The purpose of this study was to assess the value of transthoracic fine-needle aspiration in the diagnosis of mycobacterial infection as the cause of focal lung opacities. Six hundred twelve fine-needle aspiration biopsies were performed from 1985 to 1997 in 587 patients with solitary or multiple lung opacities. Initial procedures, including sputum analysis and bronchoscopy, had been nondiagnostic. Fluoroscopic or computed tomography guidance was used, and a pathologist was present. A diagnosis of mycobacterial infection was established when acid-fast bacilli were demonstrated in the aspirate. In 487 patients, a malignant cause was confirmed, and six other patients had carcinoid tumor. Of 94 nonmalignant opacities, 24 (26%) were determined to have a mycobacterial cause. Fine-needle aspiration biopsy detected acid-fast bacilli in 15 of 24 cases (sensitivity, 62.5%; specificity, 100%). Radiologic findings included upper lobe involvement (17 of 24 cases), single opacities (12 of 24 cases), satellite nodules (4 of 12 cases with single opacities), irregular borders (19 of 24), eccentric calcification (2 of 24), and cavitation (8 of 24). The authors conclude that fine-needle aspiration biopsy must be processed for acid-fast bacilli when nonmalignant cytologic findings result, even if the results of sputum smears, cultures, and bronchoscopy are negative.

Adolescent↗

Mesophilic Aeromonas strains from different serogroups: the influence of growth temperature and osmolarity on lipopolysaccharide and virulence.

Growth of mesophilic Aeromonas sp. strains from serogroups O:13, O:33 and O:44 at different temperatures and osmolarity resulted in changes in the lipopolysaccharide (LPS) and virulence of the strains tested, as we had previously reported for strains from serogroup O:34. The effect of osmolarity could be observed when the cells grew at 37 degrees C but not at 20 degrees C. Purified LPS from cells cultivated at 20 degrees C (high or low osmolarity) or at 37 degrees C at high osmolarity was smooth, whereas the LPS extracted from the cells cultivated on low osmolarity was rough. The smooth strains were resistant to the bactericidal activity of non-immune serum, while the rough strains were sensitive and showed better adhesion to Hep-2 cells than the rough strains. Furthermore, the smooth strains were more virulent for fish and mice than the rough strains. For mesophilic Aeromonas sp. strains from serogroups O:1 to O:44, these changes were not observed, except for serogroups O:13, O:33, O:34 and O:44.

Aeromonas↗