51 research outputs found
A recurrent mitochondrial p.Trp22Arg NDUFB3 variant causes a distinctive facial appearance, short stature and a mild biochemical and clinical phenotype
Background Isolated Complex I deficiency is the most common paediatric mitochondrial disease presentation, associated with poor prognosis and high mortality. Complex I comprises 44 structural subunits with at least 10 ancillary proteins; mutations in 29 of these have so far been associated with mitochondrial disease but there are limited genotype-phenotype correlations to guide clinicians to the correct genetic diagnosis.
Methods Patients were analysed by whole-exome sequencing, targeted capture or candidate gene sequencing. Clinical phenotyping of affected individuals was performed.
Results We identified a cohort of 10 patients from 8 families (7 families are of unrelated Irish ancestry) all of whom have short stature (C, p.Trp22Arg NDUFB3 variant. Two sibs presented with primary short stature without obvious metabolic dysfunction. Analysis of skeletal muscle from three patients confirmed a defect in Complex I assembly.
Conclusions Our report highlights that the long-term prognosis related to the p.Trp22Arg NDUFB3 mutation can be good, even for some patients presenting in acute metabolic crisis with evidence of an isolated Complex I deficiency in muscle. Recognition of the distinctive facial features—particularly when associated with markers of mitochondrial dysfunction and/or Irish ancestry—should suggest screening for the p.Trp22Arg NDUFB3 mutation to establish a genetic diagnosis, circumventing the requirement of muscle biopsy to direct genetic investigations
Recombinant forms of Leishmania amazonensis excreted/secreted promastigote surface antigen (PSA) induce protective immune responses in dogs
International audiencePreventive vaccination is a highly promising strategy for interrupting leishmaniasis transmission that can, additionally, contribute to elimination. A vaccine formulation based on naturally excreted secreted (ES) antigens was prepared from L. infantum promastigote culture supernatant. This vaccine achieved successful results in Phase III trials and was licensed and marketed as CaniLeish. We recently showed that newly identified ES promastigote surface antigen (PSA), from both viable promastigotes and axenically-grown amastigotes, represented the major constituent and the highly immunogenic antigen of L. infantum and L. amazonensis ES products. We report here that three immunizations with either the recombi-nant ES LaPSA-38S (rPSA) or its carboxy terminal part LaPSA-12S (Cter-rPSA), combined with QA-21 as adjuvant, confer high levels of protection in naive L. infantum-infected Beagle dogs, as checked by bone marrow parasite absence in respectively 78.8% and 80% of vaccinated dogs at 6 months post-challenge. The parasite burden in infected vaccinated dogs was significantly reduced compared to placebo group, as measured by q-PCR. Moreover, our results reveal humoral and cellular immune response clear-cut differences between vaccinated and control dogs. An early increase in specific IgG2 antibodies was observed in rPSA/QA-21-and Cter-rPSA/QA-21-immunized dogs only. They were found functionally active in vitro and were highly correlated with vaccine protection. In vaccinated protected dogs, IFN-γ and NO productions, as well as anti-leishmanial macrophage activity, were increased. These data strongly suggest that ES PSA or its carboxy-terminal part, in recom-binant forms, induce protection in a canine model of zoonotic visceral leishmaniasis by inducing a Th1-dominant immune response and an appropriate specific antibody response. These data suggest that they could be considered as important active components in vaccine candidates
Granzyme B-induced mitochondrial ROS are required for apoptosis
Caspases and the cytotoxic lymphocyte protease granzyme B (GB) induce reactive oxygen species (ROS) formation, loss of transmembrane potential and mitochondrial outer membrane permeabilization (MOMP). Whether ROS are required for GB-mediated apoptosis and how GB induces ROS is unclear. Here, we found that GB induces cell death in an ROS-dependent manner, independently of caspases and MOMP. GB triggers ROS increase in target cell by directly attacking the mitochondria to cleave NDUFV1, NDUFS1 and NDUFS2 subunits of the NADH: ubiquinone oxidoreductase complex I inside mitochondria. This leads to mitocentric ROS production, loss of complex I and III activity, disorganization of the respiratory chain, impaired mitochondrial respiration and loss of the mitochondrial cristae junctions. Furthermore, we have also found that GB-induced mitocentric ROS are necessary for optimal apoptogenic factor release, rapid DNA fragmentation and lysosomal rupture. Interestingly, scavenging the ROS delays and reduces many of the features of GB-induced death. Consequently, GB-induced ROS significantly promote apoptosis
In Vitro Evaluation of a Soluble Leishmania Promastigote Surface Antigen as a Potential Vaccine Candidate against Human Leishmaniasis
International audiencePSA (Promastigote Surface Antigen) belongs to a family of membrane-bound and secreted proteins present in severalLeishmania (L.) species. PSA is recognized by human Th1 cells and provides a high degree of protection in vaccinated mice.We evaluated humoral and cellular immune responses induced by a L. amazonensis PSA protein (LaPSA-38S) produced in aL. tarentolae expression system. This was done in individuals cured of cutaneous leishmaniasis due to L. major (CCLm) or L.braziliensis (CCLb) or visceral leishmaniasis due to L. donovani (CVLd) and in healthy individuals. Healthy individuals weresubdivided into immune (HHR-Lm and HHR-Li: Healthy High Responders living in an endemic area for L. major or L. infantuminfection) or non immune/naive individuals (HLR: Healthy Low Responders), depending on whether they produce high orlow levels of IFN-c in response to Leishmania soluble antigen. Low levels of total IgG antibodies to LaPSA-38S were detectedin sera from the studied groups. Interestingly, LaPSA-38S induced specific and significant levels of IFN-c, granzyme B and IL-10 in CCLm, HHR-Lm and HHR-Li groups, with HHR-Li group producing TNF-a in more. No significant cytokine response wasobserved in individuals immune to L. braziliensis or L. donovani infection. Phenotypic analysis showed a significant increasein CD4+ T cells producing IFN-c after LaPSA-38S stimulation, in CCLm. A high positive correlation was observed between thepercentage of IFN-c-producing CD4+ T cells and the released IFN-c. We showed that the LaPSA-38S protein was able toinduce a mixed Th1 and Th2/Treg cytokine response in individuals with immunity to L. major or L. infantum infectionindicating that it may be exploited as a vaccine candidate. We also showed, to our knowledge for the first time, the capacityof Leishmania PSA protein to induce granzyme B production in humans with immunity to L. major and L. infantum infectio
Low incidence of SARS-CoV-2, risk factors of mortality and the course of illness in the French national cohort of dialysis patients
Eletroforese em papel das proteÃnas do lÃqüido cefalorraquidiano: II. Principais resultados registrados na literatura
Prevalencia de metalo-β-lactamasas en Pseudomonas aeruginosa resistentes a carbapenemes en un Hospital Universitario de Buenos Aires
Se estudiaron 91 aislamientos de Pseudomonas aeruginosa resistentes a carbapenemes con el objetivo de conocer la prevalencia de metalo-β-lactamasas y evaluar la habilidad del ensayo de inhibición empleando discos de EDTA (1 µmol) en su detección. Se determinó la presencia de carbapenemasas en 10 (11%) de los aislamientos recuperados. La sensibilidad a aztreonam en los aislamientos resistentes a ambos carbapenemes resultó un buen predictor de la presencia de estas enzimas. Dichas carbapenemasas correspondieron a la enzima VIM-2 en tres de ellos y a VIM-11 en otros siete. En todos los casos los genes codificantes de estas enzimas se encontraron localizados en integrones de clase 1 seguidos corriente abajo de genes codificantes de enzimas acetilantes de antibióticos aminoglucosÃdicos. El ensayo de detección fenotÃpica de metalo-β-lactamasas empleando discos de EDTA mostró un 100% de especificidad y sensibilidad en la detección de estas enzimas en la población de Pseudomonas aeruginosa analizadas
Prevalencia de metalo-β-lactamasas en Pseudomonas aeruginosa resistentes a carbapenemes en un Hospital Universitario de Buenos Aires Prevalence of metallo-β-lactamase in carbapenem resistant Pseudomonas aeruginosa at an University Hospital of Buenos Aires City
Se estudiaron 91 aislamientos de Pseudomonas aeruginosa resistentes a carbapenemes con el objetivo de conocer la prevalencia de metalo-β-lactamasas y evaluar la habilidad del ensayo de inhibición empleando discos de EDTA (1 µmol) en su detección. Se determinó la presencia de carbapenemasas en 10 (11%) de los aislamientos recuperados. La sensibilidad a aztreonam en los aislamientos resistentes a ambos carbapenemes resultó un buen predictor de la presencia de estas enzimas. Dichas carbapenemasas correspondieron a la enzima VIM-2 en tres de ellos y a VIM-11 en otros siete. En todos los casos los genes codificantes de estas enzimas se encontraron localizados en integrones de clase 1 seguidos corriente abajo de genes codificantes de enzimas acetilantes de antibióticos aminoglucosÃdicos. El ensayo de detección fenotÃpica de metalo-β-lactamasas empleando discos de EDTA mostró un 100% de especificidad y sensibilidad en la detección de estas enzimas en la población de Pseudomonas aeruginosa analizadas.<br>The present study was conducted to estimate the prevalence of metallo-β-lactamases in 91 consecutive carbapenem resistant Pseudomonas aeruginosa isolates, recovered from inpatients at Hospital de ClÃnicas in Buenos Aires. Both, phenotypic and genotypic methods detected the presence of carbapenemases in 10 (11%) isolates, corresponding to VIM-11 in 7/10 and VIM-2 in the others. Codifying genes were all included in class 1 integrons, upstream genes coding for aminoglycoside modifying enzymes. One hundred percent sensitivity and specificity was achieved by the metallo-β-lactamases phenotypic screening method using EDTA (1 µmol) disks in the Pseudomonas aeruginosa isolates included in this study. Sensitivity to aztreonam in carbapenem resistant isolates was suspicious of the presence of these enzymes
Modification of sunflower oil quality by seed-specific expression of a heterologous Delta 9-stearoyl-(acyl carrier protein) desaturase gene
The coding sequence of Delta9-stearoyl-(acyl carrier protein) desaturase from Ricinus communis was introduced into sunflower. under the control of seed-specific promoter and terminator sequences of the late embryogenesis abundant gene from sunflower. Hads10. Two independent primary transformants contained three and six copies of the T-DNA. as demonstrated by hybridization using nptII as a probe. The transgene proved genetically stable and was transmitted as a Mendelian trait. Transcript analysis of the heterologous Delta9-stearoyl-(acyl carrier protein) desaturase under control of the Hads10 promoter verified tissue-specific expression in the developing embryos and not in the leaves. Fatty acid composition of the seed oil was followed over five generations under greenhouse and open field conditions. Some of the transgenic lines produced oil with a significantly reduced stearic acid content compared with non-transformed plants under greenhouse and field conditions, However. additional studies need to be performed to assess whether or not physiologically stable lines can be developed from these transgenic lines
- …