Informe especial: Coberturas de vacunación en residentes de la CABA en contexto Covid-19: 2 de octubre de 2020 / Special report: Vaccination coverage in CABA residents in context of Covid19

La vacunación es considerada una actividad esencial durante la pandemia de COVID-19 y se han desarrollado diferentes estrategias para el sostenimiento de la vacunación en el contexto actual, facilitar el acceso a través de la adaptación y reorganización de los servicios de salud, el no requerimiento de permisos de circulación para la vacunación, vacunación en instituciones fuera de salud, así como la elaboración de recomendaciones para realizar la vacunación de manera segura protegiendo tanto al vacunador como la persona a vacunar, entre otros. Con el objetivo de realizar la medición del impacto en las actividades de vacunación, se realiza el análisis comparativo de las vacunas aplicadas durante el primer semestre de los años 2018-2020, con la información recibida en el nivel central del Programa de Inmunizaciones. Se excluye del presente análisis la información referida a la vacuna antigripal ya que tiene una modalidad diferente de aplicación. (AU)

Vigilancia de eventos supuestamente atribuidos a la vacunación o inmunización (ESAVI) durante el año 2017 / Surveillance of events supposedly attributed to vaccination or immunization (ESAVI) during 2017

Los Eventos Supuestamente Atribuidos a la Vacunación o Inmunización o ESAVI se definen como todo cuadro clínico que aparece luego de la administración de una vacuna y que supuestamente pueda atribuirse a la misma. Incluye los errores programáticos relacionados con la vacunación. Un ESAVI grave es todo aquel evento que resulte en hospitalización o fallecimiento. Es importante mencionar que un ESAVI, si bien denota una asociación temporal, no implica necesariamente una relación de causa y efecto. La causalidad entre el evento y la vacunación se determinará mediante la investigación del caso. La información aquí presentada surge del análisis de la base de datos de ESAVI del Programa de Inmunizaciones de la Ciudad de Buenos Aires, alimentada por las notificaciones realizadas por efectores públicos y privados de la ciudad. Se incluyen residentes y no residentes de la ciudad, sin realizar distinción entre ellos. Para calcular las tasas se utilizó como denominador las dosis aplicadas en 2017 en la Ciudad de Buenos Aires, tanto a residentes como no residentes. Se cuenta con datos de aquellas vacunas incluidas en el Calendario Nacional de Vacunación del sector público, de la seguridad social y privado. (AU)

[Active surveillance of adverse effects of Pandemrix vaccine to prevent influenza A(H1N1) in Cuba]. / Vigilancia activa de eventos adversos a la vacuna Pandemrix para prevenir la influenza AH1N1 en Cuba.

INTRODUCTION: in April 2009, a new virus was identified in Mexico and North America as the cause of a respiratory disease. The virus quickly spread over other countries. On June 11, 2009 the World Health Organization (WHO) reported cases in 74 countries and territories located in 2 of its regions. The high sustained transmission of this virus worldwide led to establish the phase 6 or the pandemic phase, indicating that the situation had to do with spreading rather than increased severity. OBJECTIVES: to report on already known or new events after the administration of vaccine A(H1N1) called Pandemrix, to identify the most frequent events occurred in pregnant women and to research into the associated severe events. METHODS: a prospective descriptive study was designed to characterize the adverse effects of Pandemrix reported across the country from April 1st to June 30th, 2010. A total of 1,123,526 people were vaccinated in which 100% of pregnant women were included. RESULTS: active surveillance nationwide reported 5 763 signs and symptoms detected in 3 401 people (615 reports from pregnant women). The overall rate of reports was 302.7 x 100 000 doses administered. Adverse events such as fever, headache, pain, swelling and redness at the injection site, malaise, arthralgia, allergic reactions, nausea and vomiting were reported as common symptoms. These 10 symptoms and signs accounted for 79.1% of all the reported events. A total number of 80 317 pregnant women were vaccinated of whom 615 reported adverse effects, accounting for 0.8 % of the vaccinated pregnant women. Fever was the most notified symptom in children (193) followed by local reactions at the injection site (23), vomiting (20), arthralgia (17), headache (11), malaise (10) and high fever-related seizures (6). Eight events were analyzed as severe. CONCLUSIONS: the administration of the vaccine was related to 3 events, unrelated to other 3 events and 2 were classified as inconclusive (3 miscarriages). No deaths were reported. The capacity of the Cuban Health System for the administration of this vaccine with active surveillance in a short period of time was proved. Data from monitoring of events that were supposedly attributable to vaccination or immunization did not notify any unusual event. Therefore, no safety problem is associated to the Pandemrix vaccine.

Biochemical characterization of mapmodulin, a protein that binds microtubule-associated proteins.

Mapmodulin is a 31-kDa protein that stimulates the microtubule- and dynein-dependent localization of Golgi complexes in semi-intact Chinese hamster ovary cells. We have shown previously that it binds the microtubule binding domains of the microtubule-associated proteins, MAP2, MAP4, and tau. We also showed that mapmodulin is identical to a protein named PHAPI (Vaesen, M., Barnikol-Watanabe, S. , Götz, H., Awni, L.A., Cole, T., Zimmermann, B., Kratzin, H.D. and Hilschmann, N. (1994) Biol. Chem. Hoppe-Seyler 375, 113-126). We report here that mapmodulin is a phosphoprotein that is predominantly cytosolic but is also found peripherally associated with endoplasmic reticulum and Golgi membranes in mammalian cells. The protein occurs as a trimer in cytosol, and phosphorylation is required for its microtubule-associated protein-binding activity. Heat treatment of nonphosphorylated mapmodulin can render it competent for binding to microtubule-associated proteins, suggesting that phosphorylation induces a conformational change in mapmodulin. Finally, despite identity in polypeptide sequence with a protein reported to act as an inhibitor of protein phosphatase 2A, native mapmodulin was not able to inhibit protein phosphatase 2A in Chinese hamster ovary cell cytosol.

A novel assay reveals a role for soluble N-ethylmaleimide-sensitive fusion attachment protein in mannose 6-phosphate receptor transport from endosomes to the trans Golgi network.

Soluble N-ethylmaleimide-sensitive fusion protein (NSF) attachment protein (alpha-SNAP) is a soluble protein that enables the NSF ATPase to associate with membranes and facilitate membrane trafficking events. Although NSF and alpha-SNAP have been shown to be required for many membrane transport processes, their role in the transport of mannose 6-phosphate receptors from endosomes to the trans Golgi network was not established. We present here a novel in vitro assay that monitors the transport of cation-dependent mannose 6-phosphate receptors between endosomes and the trans Golgi network. The assay relies on the trans Golgi network localization of tyrosine sulfotransferase and monitors transport of mannose 6-phosphate receptors engineered to contain a consensus sequence for modification by this enzyme. Using this new assay we show that alpha-SNAP strongly stimulates transport in reactions containing limiting amounts of cytosol. Together with alpha-SNAP, NSF can increase the extent of transport. These data show that alpha-SNAP, a soluble component of the SNAP receptor machinery, facilitates transport from endosomes to the trans Golgi network.

A tandem array of Sp-1 sites and a reverse initiator element are both required for synergistic transcriptional activation of the T-cell-specific MAL gene.

We have characterized the three cis elements responsible for promoter strength present in the 5'-flanking proximal region of MAL, a human T-cell-specific gene encoding a proteolipid protein present in detergent-insoluble complexes of high molecular weight. The first element consisted of an initiator sequence that, curiously, was present in reverse orientation compared to that of the standard initiator elements. The other two elements were contained in a region of 126 bp upstream of the mRNA initiation site, and consisted of a tandem array of one GC box and one GA box. The GC box corresponds to a consensus site for the nuclear factor Sp1, whereas the GA box deviates from this consensus, although it was able to compete for the binding of Sp1 in vitro and to respond to trans-activation by Sp1 in vivo. This simple promoter lacks an apparent TATA box and lost more than 99% of its activity when a fragment of 60 bp containing the GC and GA boxes was deleted. A synergistic effect on transcriptional activation was observed in the presence, but not in the absence, of the initiator element when both GC and GA boxes were present.

The MAL proteolipid is a component of the detergent-insoluble membrane subdomains of human T-lymphocytes.

The human mal gene, identified during a search for cDNAs selectively expressed during T-cell development, encodes a highly hydrophobic protein belonging to a group of proteins, termed proteolipids, characterized by their unusual property of being soluble in organic solvents used to extract cell lipids. To study the localization of the MAL protein we have prepared stable transfectants expressing the MAL protein tagged with a c-myc epitope (MAL/c-myc) using human epithelial A-498 cells. Immunofluorescence analysis suggested that MAL/c-myc is localized mainly to cholesterol-enriched structures with a post-Golgi location and, at low levels, in early endosomes. Moreover, extraction of A-498 cell membranes with Triton X-100 (TX100) and fractionation by centrifugation to equilibrium in sucrose gradients demonstrated the presence of MAL/c-myc in the detergent-insoluble buoyant fraction, known to be enriched in glycolipids and cholesterol. To compare the behaviour of MAL in T-cells with that in epithelial A-498 cells, we prepared stably transfected cells expressing MAL/c-myc using human Jurkat T-cells. When TX100 extracts from Jurkat cells were subjected to centrifugation to equilibrium in sucrose gradients we found MAL exclusively in the floating fractions, together with molecules characteristic of the T-cell insoluble complexes, such as the tyrosine kinase p56lck, the glycosylphosphatidylinositol-anchored protein CD59 and the ganglioside GM1. These results, taken together, indicate that the MAL proteolipid is a component of the detergent-resistant membrane microdomains present in T-lymphocytes, and suggest that MAL might play a role in modulating the function of these microdomains during T-cell differentiation.

Rab7 and Rab9 are recruited onto late endosomes by biochemically distinguishable processes.

Rab GTPases are localized to the surfaces of distinct membrane-bound organelles and function in transport vesicle docking and/or fusion. Prenylated Rab9, bound to GDP dissociation inhibitor-alpha, can be recruited selectively onto a membrane fraction enriched in late endosomes; this process is accompanied by nucleotide exchange. We used this system to address whether each Rab uses a distinct machinery to associate with its cognate organelle. Purified, prenylated Rab1B, Rab7, and Rab9 proteins were each reconstituted as stoichiometric complexes with purified GDP dissociation inhibitor-alpha, and their recruitment onto endosome- or ER-enriched membrane fractions was quantified. The two late endosomal proteins, Rab9 and Rab7, were each recruited onto endosome membranes with approximate apparent Km values of 9 and 22 nM, respectively. However, while control Rab9.GDP dissociation inhibitor-alpha complexes inhibited the initial rate of myc-tagged Rab9 recruitment with an apparent Ki of approximately 9 nM, Rab7 complexes inhibited this process much less effectively (apparent Ki approximately 112 nM). Similarly, complexes of the endoplasmic reticulum-localized Rab1B protein were even less potent than Rab7 complexes (apparent Ki approximately 405 nm). Rab9 complexes inhibited Rab7 recruitment with the same low efficacy as Rab7 complexes inhibited Rab9 recruitment. These experiments distinguish, biochemically, the recruitment of different Rab proteins onto a single class of organelle. Since Rab7 and Rab9 are both localized at least in large part, to late endosomes, this suggests that a single organelle may bear multiple Rab recruitment machines.

Alternative splicing of human T-cell-specific MAL mRNA and its correlation with the exon/intron organization of the gene.

Sequence analysis of the T-cell-specific MAL gene revealed four exons, each encoding a hydrophobic, presumably membrane-associated, segment and its adjacent hydrophilic sequence. Amplification by the polymerase chain reaction of cDNA from different T-cell samples indicated the existence of four different forms of MAL mRNA, termed MAL-a, -b, -c, and -d, that arise from differential usage of exons II and/or III. As the three introns were located between complete codons, the reading frame was maintained in all the transcripts. A model resembling the structures postulated for different proteolipid proteins is proposed for the protein encoded by each alternative mRNA species.

Genomic structure and subcellular localization of MAL, a human T-cell-specific proteolipid protein.

Genomic DNA clones containing the T-cell-specific human MAL gene were isolated. Restriction and sequence analysis revealed four exons and three introns. Each hydrophobic segment of MAL together with its adjacent hydrophilic sequence correlates closely with one exon of the gene. RNase protection analysis revealed that the previously described MAL mRNA, which contains the sequences present in the four exons, is the mRNA species predominant in T-cells. A remarkable similarity was found between the hydrophobicity pattern of MAL and those of the peripheral membrane protein 22 (PMP-22) and the 16-kDa proteolipid of vacuolar H(+)-ATPase. Direct evidence supporting that MAL is a proteolipid was obtained by extracting bacterial lysates expressing recombinant MAL protein with lipophilic solvents used to extract lipids. The use of two different antibodies raised against distinct peptides from the MAL molecule has allowed the localization of MAL in the endoplasmic reticulum of T-cells. This subcellular localization is in agreement with the presence of a RWKSS motif in the COOH-terminal tail of MAL, next to its last putative transmembrane domain, that fits with one of the consensus sequences (RXKXX) for residency in the endoplasmic reticulum for transmembrane proteins. A possible function for MAL protein in T-cells is discussed based on its subcellular localization and the unique lipid-like properties of the proteolipid proteins.