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Biomedical subjects

O Lopez

Publications and source records attributed to O Lopez.

At least 37 records · Page 2Linked to original sources

Solubilization of phosphatidylcholine liposomes by the amphoteric surfactant dodecyl betaine.

The interaction of the amphoteric surfactant N-dodecyl-N,N-dimethylbetaine (C12-Bet) with phosphatidylcholine (PC) liposomes was investigated. Permeability alterations were detected as a change in 5(6)-carboxyfluorescein (CF) released from the interior of vesicles and bilayer solubilization as a decrease in the static light-scattering (SLS) of the system. At sublytic level a initial maximum in the bilayer/water partitioning (K) followed by an abrupt decrease of this parameter occurred as the surfactant to lipid molar ratio (Re) rose. At lytic level a direct dependence was established between both parameters. The fact that the free surfactant concentration at sublytic and lytic levels showed values lower than and similar to its critical micelle concentration indicates that permeability alterations and solubilization were determined, respectively, by the action of surfactant monomer and by the formation of mixed micelles. A direct correlation occurred in the initial interaction steps (up to 50% CF release) between the growth of vesicles their fluidity and Re. A similar direct dependence was established during solubilization (up to 30% SLS) between the fall in both the surfactant-lipid aggregate size, the SLS of the system and Re. This surfactant showed higher capacity to solubilize PC liposomes than that reported by the commonly used non-ionic surfactants octyl glucoside and Triton X-100 and by the anionic one sodium dodecyl sulfate.

Betaine↗

Influence of the level of ceramides in the permeability of stratum corneum lipid liposomes caused by sodium dodecyl sulfate.

The role played by the ceramides in the sublytic interactions of sodium dodecyl sulfate (SDS) with liposomes modeling the stratum corneum (SC) lipid composition was studied. The surfactant/lipid molar ratios (Re) and the bilayer/aqueous phase partition coefficients (k) were determined by monitoring the changes in the fluorescence intensity of liposomes due to the 5(6) carboxyfluorescein (CF) released from the interior of vesicles. The presence in liposomes of higher and lower ceramide proportions than that existing in the SC lipids led to a fall and to a rise in the sublytic activity of SDS on these structures. However, the SDS partitioning into liposomes (or affinity with these bilayer structures) increased as the proportion of Cer increased up to achieve almost a constant value for a Cer proportion similar to that in the SC lipids (about 40%). Thus, at low Cer proportions the ability of SDS molecules to alter these bilayer structures was higher than that for liposomes approximating the SC lipid composition despite their reduced partitioning into liposomes. These findings are in agreement with the recently reported dependencies of the level of ceramides in skin lipids and function barrier abnormalities and could explain in part these dependencies. The fact that the free surfactant concentration needed to achieve the two interaction levels investigated was lower than the surfactant critical micellar concentration (CMC) indicates that permeability alterations were mainly ruled by the action of surfactant monomers, regardless of the liposome lipid composition.

Ceramides↗

A single VH family and long CDR3s are the targets for hypermutation in bovine immunoglobulin heavy chains.

Bovine immunoglobulins are made from genes belonging to a small family of closely related VH genes. In this respect cattle resemble all species of domesticated mammals, which also use one VH family. The family, named BoVH1, is homologous to the mouse Q52 family, and there are no more than 20 genes of this family in the bovine genome. Another feature of bovine heavy chains is the use of long CDR3s, which have an average of 21 codons. It seems that there are several families of long, closely related D genes rich in glycine and tyrosine responsible for this length. Sequences described as targets for mutations in other species can be found in CDR1, CDR2, and the putative D genes. The mutation mechanism starts at some point between late fetal stage and birth and seems to be antigen independent. Diversity seems to be generated by hypermutation, although other mechanisms cannot be discounted at this time. Contrary to humans and mice, which have several VH gene families comprising more than 100 genes, cattle use only a few genes and long CDR3s followed by somatic mutation to generate the necessary diversity to recognize the universe of antigens they will encounter during their life.

Amino Acid Sequence↗

Genetic evidence for 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) as a negative effector of cytochrome terminal oxidase cbb3 production in Rhizobium etli.

A Rhizobium etli Tn5mob-induced mutant (CFN035) exhibits an enhanced capacity to oxidize N,N,N',N', tetramethyl-p -phenylenediamine (TMPD), a presumptive indicator of elevated cytochrome c terminal oxidase activity. Sequencing of the mutated gene in CFN035 revealed that it codes for the amidophosphoribosyl transferase enzyme (PurF) that catalyzes the first step in the purine biosynthetic pathway. Two c-type cytochromes with molecular weights of 32 and 27 kDa were produced in strain CFN035, which also produced a novel CO-reactive cytochrome (absorbance trough at 553 nm), in contrast to strain CE3 which produced a single 32 kDa c-type protein and did not produce the 553 nm CO-reactive cytochrome. A wild-type R. etli strain that expresses the Bradyrhizobium japonicum fixNOQP genes, which code for the symbiotic cytochrome terminal oxidase cbb3, produced similar absorbance spectra (a trough at 553 nm in CO-difference spectra) and two c-type proteins similar in size to those of strain CFN035, suggesting that CFN035 also produces the cbb3 terminal oxidase. The expression of a R. etli fixN-lacZ gene fusion was measured in several R. etli mutants affected in different steps of the purine biosynthetic pathway. Our analysis showed that purF, purD, purQ, purL, purY, purK and purE mutants expressed three-fold higher levels of the fixNOQP operon than the wild-type strain. The derepressed expression of fixN was not observed in a purH mutant. The purH gene product catalyzes the conversion of 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) to 5-form-aminoimidazole-4-carboxamide ribonucleotide (FAICAR) and inosine. Supplementation with AICA riboside lowered the levels of fixN expression in the purF mutants. These data are consistent with the possibility that AICAR, or a closely related metabolite, is a negative effector of the production of the symbiotic terminal oxidase cbb3 in R. etli.

Amidophosphoribosyltransferase↗

Gag-specific cytotoxic responses to HIV type 1 are associated with a decreased risk of progression to AIDS-related complex or AIDS.

The duration of human immunodeficiency virus (HIV-1) infection prior to the development of AIDS is variable, and for most patients the exact time of infection is not known. A group of 38 HIV-1-infected subjects was tested while asymptomatic for comparative cytotoxic lymphocyte responses to the Gag and envelope antigens of HIV-1. Twenty of the 38 patients had no detectable primary cytotoxic T lymphocyte (CTL) response to Gag, and this was associated with a relative risk of 1.89 for progression to ARC or AIDS during the subsequent 3 to 40 months of observation when compared with patients who had Gag-specific CTL activity at the beginning of the observation period. In contrast, no significant association was observed between envelope-specific cytotoxic activity and disease progression. Other patient characteristics, including CD4+ T lymphocyte counts and antibody levels to the p24gag protein, measured at the start of observation, did not correlate with disease progression during the observation period. This suggests that the anti-Gag CTL response may be protective during HIV-1 infection.

Acquired Immunodeficiency Syndrome↗

Cloning and molecular characterization of two genes encoding adhesion proteins involved in Trichomonas vaginalis cytoadherence.

Cytoadherence to the vaginal epithelium is a critical step in infection by the eukaryotic flagellate Trichomonas vaginalis. Four trichomonad surface proteins (AP65, AP51, AP33 and AP23) mediate cytoadherence. The cDNA encoding the AP65 adhesin was isolated from a phagemid cDNA expression library by screening with antiserum and monoclonal antibody (mAb) raised against the purified trichomonad AP65 protein. Two clones, F11.2 and F11.5, coded for immuno-crossreactive recombinant proteins that possessed functional properties equal to the T. vaginalis AP65 adhesin. Analysis of full-length sequences corresponding to the F11.2 and F11.5 cDNAs revealed that both contained 1701-base open reading frames (ORFs) that encoded proteins of 63 281 daltons and 83 087 daltons, respectively. Comparison of the full-length sequences showed 87% identity at the nucleotide level and 91% identity at the protein level. Restriction-enzyme mapping and Southern analysis reaffirmed the distinctness of the F11.2 and F11.5 cDNAs, indicating that two different AP65 genes (now called ap65-1 and ap65-2) are present in the T. vaginalis genome in at least two copies each. Northern analysis detected high levels of transcript of approximately 1.8 kb for both ap65-1 and ap65-2 genes in trichomonads grown only in high-iron medium, confirming the transcriptional regulation of adhesin synthesis by iron. Homology searches revealed significant similarity (38% amino acid identity and 54% nucleotide identity) to malic enzymes. However, purified malic enzyme and mAb to AP65 crossreactive with malic enzyme neither inhibited cytoadherence of T. vaginalis to host cells nor prevented binding of the trichomonad AP65 to HeLa cells in a ligand assay.

Amino Acid Sequence↗

[Oxidative stress in 29 HIV seropositive patients. Two-year results of a double-blind trial of diethyldithiocarbamate versus placebo].

Among 29 seropositive subjects who had participated in the HIV 87 therapeutic trial (Mérieux laboratories), the oxidative stress was evaluated at 24 months in 16 treated with diethyldithiocarbamate (dithiocarb) and in 13 who had received the placebo. No significant difference was found between these two groups, whereas the existence of an oxidative stress has been confirmed in seropositive subjects compared with controls.

Adult↗

[Anonymous and free-of-charge detection of human immunodeficiency virus infection. 20-months' experience at a center].

The results of 20 months' activity of the anonymous and free-of-charge detection centre of the Pitié-Salpêtrière hospital group, Paris, concerning human immunodeficiency virus infection (HIV) are presented. During that period, 3,480 persons consulted and 3,332 anonymous questionnaires were filled and returned: 20.5 percent of the subjects were homosexuals, 6.5 percent were drug-addicts and 73 percent were non drug-addict heterosexuals; 31 percent used condoms. A total of 3,398 blood samples were collected; 232 sera were positive or undetermined for HIV-1 and/or HIV-2 by the ELISA method; 132 Western Blot tests confirmed the positivity for HIV-1 but not for HIV-2. The overall serum positivity was 4 percent; 18.3 percent of drug-addicts, 9.5 percent of homosexuals and 0.9 percent of heterosexuals were HIV positive. Among seropositive subjects, 51 percent were homosexuals, 27 percent were drug-addicts, 4 percent were homosexual drug-addicts and 18 percent were heterosexuals (43 percent of these had had multiple partners); condoms were used by 59 percent of HIV positive subjects. The percentage of HIV positive subjects in our series was lower than that estimated in populations at risk (drug-addicts 50 percent, homosexuals 32 percent); it was similar to the percentages found in other detection centres (5 to 6 percent). Most seropositive patients belong to the category of persons who are the first to be struck by HIV. The heterosexual population is relatively spared, but most of the recent seroconversions have occurred in this group.

Acquired Immunodeficiency Syndrome↗

Alzheimer's disease, cerebral amyloid angiopathy, and dementia of acute onset.

The coexistence of cerebral amyloid angiopathy (CAA) and Alzheimer's disease (AD) should be considered in the differential diagnosis of cases with acute onset of dementia when other causes have been excluded. We report clinical and neuropathological findings in a 78-year-old man who developed dementia of acute onset with an apparent rapid course three months before his death. Postmortem microscopic examination of the brain revealed senile (neuritic) plaques and neurofibrillary tangles in the hippocampus and cerebral cortex. CAA affected vessels of the neocortex and leptomeninges, most severely in the frontal and parietal areas.

Aged↗

New refractive method for laser thermal keratoplasty with the Co:MgF2 laser.

We have observed corneal curvature changes from laser thermal keratoplasty with a Co:MgF2 laser. We studied corneal curvature changes in rabbits and have identified specific treatment patterns and laser parameters that can correct myopia and astigmatism. These corneal changes, some as large as 8 diopters, have been stable for at least one year, and slitlamp examination demonstrates clear central corneas with normal appearance.

Animals↗

Primary cytotoxicity against the envelope glycoprotein of human immunodeficiency virus-1: evidence for antibody-dependent cellular cytotoxicity in vivo.

Fresh peripheral blood mononuclear cells from human immunodeficiency virus-1 (HIV-1) seropositive donors can lyse target cells expressing the envelope glycoprotein in vitro. In most cases, this antigen-specific lysis is not mediated by T lymphocytes. Lymphoblastoid cell lines infected with recombinant vaccinia viruses expressing different forms of the envelope protein of HIV-1 were used as target cells in chromium-release assays of primary cytotoxic effector cells and of antibody-dependent cellular cytotoxicity (ADCC). By depleting effector cells of CD16+ lymphocytes, or by blocking target cell lysis with an anti-human IgG serum, primary env-specific lysis was found to be due to ADCC, the effector cells being armed in vivo with specific, cytophilic antibodies. This phenomenon is dependent on cell surface expression of the envelope protein and is directed against both gp120 and gp41. Human HIV-1 antibody-positive sera are able to mediate ADCC against HIV-infected CD4+ T lymphocytes, suggesting a possible role of ADCC in the natural infection.

Antibody-Dependent Cell Cytotoxicity↗

Human immunodeficiency virus-specific cytotoxic responses of seropositive individuals: distinct types of effector cells mediate killing of targets expressing gag and env proteins.

By using target cells that expressed isolated env, gag, p27nef, or p23vif molecules introduced by recombinant vaccinia viruses containing genes encoding these polypeptides, it was possible to identify env, gag, p27nef, and p23vif as cytolytic target antigens for freshly isolated blood cells from human immunodeficiency virus 1 (HIV-1) seropositive patients. Most of the patients tested (95%) manifested a specific cytotoxic activity against vaccinia virus-env-infected target cells. The env-specific cytotoxic activity was not restricted by the major histocompatibility complex and was not mediated by T lymphocytes, as shown by the absence of blocking effect with an anti-CD3 monoclonal antibody and by the inefficiency of CD3+, CD8+, or CD4+ and CD8+ depletion to reduce the cytotoxic activity against the env-expressing target cells. In the same conditions, the cytotoxic activity specific for gag was abrogated and gag major histocompatibility complex-restricted cytotoxic T lymphocytes were detected in 85% of the subjects tested. Therefore, in a HIV-1 seropositive subject, distinct types of effector cells mediate the lysis of target cells expressing gag and env proteins.

Antibodies, Monoclonal↗

Seizures after liver transplantation: a clinicopathologic study.

We reviewed the clinical and neuropathologic findings in 21 patients who had seizures after orthotopic liver transplants. Tonic-clonic seizures were the most common seizure type. Six patients developed status epilepticus. In 9 patients, seizures occurred within 1 week following transplantation. We found CNS lesions that were probably responsible for the occurrence of seizures in most patients; some had more than 1 finding. Neuropathologic examination revealed ischemic or hemorrhagic strokes in 18 patients, central pontine myelinolysis in 5, and CNS infections in 5. Multiple metabolic abnormalities were a contributing factor to the onset of seizures in some patients.

Adult↗