Search PubMed⌕ Search

Biomedical subjects

A Portela

Publications and source records attributed to A Portela.

At least 19 recordsLinked to original sources

Global priority setting for Cochrane systematic reviews of health promotion and public health research.

BACKGROUND: Systematic reviews of health promotion and public health interventions are increasingly being conducted to assist public policy decision making. Many intra-country initiatives have been established to conduct systematic reviews in their relevant public health areas. The Cochrane Collaboration, an international organisation established to conduct and publish systematic reviews of healthcare interventions, is committed to high quality reviews that are regularly updated, published electronically, and meeting the needs of the consumers. AIMS: To identify global priorities for Cochrane systematic reviews of public health topics. METHODS: Systematic reviews of public health interventions were identified and mapped against global health risks. Global health organisations were engaged and nominated policy-urgent titles, evidence based selection criteria were applied to set priorities. RESULTS: 26 priority systematic review titles were identified, addressing interventions such as community building activities, pre-natal and early infancy psychosocial outcomes, and improving the nutrition status of refugee and displaced populations. DISCUSSION: The 26 priority titles provide an opportunity for potential reviewers and indeed, the Cochrane Collaboration as a whole, to address the previously unmet needs of global health policy and research agencies.

Evidence-Based Medicine↗

Empowerment of women, men, families and communities: true partners for improving maternal and newborn health.

Based on the Health Promotion approach, the Making Pregnancy Safer initiative has proposed a strategic framework for working with individuals, families and communities to improve maternal and newborn health. The aims are to contribute to the empowerment of women, families and communities to increase their influence and control over maternal and newborn health, as well as to increase access to and utilization of quality skilled care. The framework has identified those strategies and interventions that target the factors known to contribute to health inequalities and poor maternal and newborn health. While empowerment is an aim of the framework, it is also considered a means. Emphasis is placed on the processes and the quality of the processes rather than just on the actions themselves. The authors in this paper would like to contribute to ongoing discussions about the 'how' of working with women, men, families and communities for improved maternal and newborn health.

Decision Making↗

Influenza A virus NEP (NS2 protein) downregulates RNA synthesis of model template RNAs.

The influenza A virus NEP (NS2) protein is an structural component of the viral particle. To investigate whether this protein has an effect on viral RNA synthesis, we examined the expression of an influenza A virus-like chloramphenicol acetyltransferase (CAT) RNA in cells synthesizing the four influenza A virus core proteins (nucleoprotein, PB1, PB2, and PA) and NEP from recombinant plasmids. Influenza A virus NEP inhibited drastically, and in a dose-dependent manner, the level of CAT expression mediated by the recombinant influenza A virus polymerase. This inhibitory effect was not observed in an analogous artificial system in which expression of a synthetic CAT RNA is mediated by the core proteins of an influenza B virus. This result ruled out the possibility that inhibition of reporter gene expression was due to a general toxic effect induced by NEP. Analysis of the virus-specific RNA species that accumulated in cells expressing the type A recombinant core proteins and NEP showed that there was an important reduction in the levels of minireplicon-derived vRNA, cRNA, and mRNA molecules. Taken together, the results obtained suggest a regulatory role for NEP during virus-specific RNA synthesis, and this finding is discussed regarding the biological implications for the virus life cycle.

Animals↗

Remission of heart failure through endoluminal repair of femoral arteriovenous fistula with the use of a covered stent.

We report the case of a 21-year-old male with high-output heart failure due to a femoral arteriovenous fistula caused by a firearm wound. A new balloon expandable stent covered with polytetrafluorethylene was implanted in the artery to occlude the arteriovenous fistula. The fistula was immediately occluded and the artery remained patent. On the following day, the patient felt much better, with no symptoms of heart failure. Additional follow-up is required to assure the usefulness of this less invasive procedure in the treatment of arteriovenous fistulas.

Adult↗

Rescue of synthetic RNAs into thogoto and influenza A virus particles using core proteins purified from Thogoto virus.

The ribonucleoprotein (RNP) complexes of Thogoto virus (THOV), a tick-borne orthomyxovirus, have been purified from detergent-lysed virions. The purified RNPs were then disrupted by centrifugation through a CsCl-glycerol gradient to obtain fractions highly enriched in nucleoprotein (NP) and virtually devoid of viral genomic RNA. When these NP-enriched fractions were incubated with a synthetic THOV-like RNA, and the mixtures were transfected into THOV-infected cells, the synthetic RNA was expressed and packaged into THOV particles. Similarly, hybrid mixtures containing purified THOV NP and influenza A virus synthetic RNAs (either a model CAT RNA or a gene encoding the viral neuraminidase), were prepared and transfected into influenza A virus-infected cells. The synthetic CAT RNA, was shown to be expressed and packaged into virus particles, and the neuraminidase gene was rescued into influenza virions. These data are discussed in terms of the similarities observed between THOV and influenza A virus and the potential application of the THOV purified proteins for rescuing synthetic genes into infectious viruses.

Animals↗

Several protein regions contribute to determine the nuclear and cytoplasmic localization of the influenza A virus nucleoprotein.

A systematic analysis was carried out to identify the amino acid signals that regulate the nucleo-cytoplasmic transport of the influenza A virus nucleoprotein (NP). The analysis involved determining the intracellular localization of eight deleted recombinant NP proteins and 14 chimeric proteins containing the green fluorescent protein fused to different NP fragments. In addition, the subcellular distribution of NP derivatives that contained specific substitutions at serine-3, which is the major phosphorylation site of the A/Victoria/3/75 NP, were analysed. From the results obtained, it is concluded that the NP contains three signals involved in nuclear accumulation and two regions that cause cytoplasmic accumulation of the fusion proteins. One of the karyophilic signals was located at the N terminus of the protein, and the data obtained suggest that the functionality of this signal can be modified by phosphorylation at serine-3. These findings are discussed in the context of the transport of influenza virus ribonucleoprotein complexes into and out of the nucleus.

Amino Acid Substitution↗

Influenza virus matrix protein is the major driving force in virus budding.

To get insights into the role played by each of the influenza A virus polypeptides in morphogenesis and virus particle assembly, the generation of virus-like particles (VLPs) has been examined in COS-1 cell cultures expressing, from recombinant plasmids, different combinations of the viral structural proteins. The presence of VLPs was examined biochemically, following centrifugation of the supernatants collected from transfected cells through sucrose cushions and immunoblotting, and by electron-microscopic analysis. It is demonstrated that the matrix (M1) protein is the only viral component which is essential for VLP formation and that the viral ribonucleoproteins are not required for virus particle formation. It is also shown that the M1 protein, when expressed alone, assembles into virus-like budding particles, which are released in the culture medium, and that the recombinant M1 protein accumulates intracellularly, forming tubular structures. All these results are discussed with regard to the roles played by the virus polypeptides during virus assembly.

Animals↗

Efficient formation of influenza virus-like particles: dependence on the expression levels of viral proteins.

It has previously been demonstrated in this laboratory that an influenza virus-like chloramphenicol acetyltransferase (CAT) RNA could be expressed in COS-1 cells that synthesized all ten influenza A virus-encoded proteins from recombinant plasmids. It was also shown that supernatant fluids harvested from these cultures contained virus-like particles (VLPs) that could deliver an enclosed CAT RNA to MDCK cells. Here, it is shown that the levels of expression of the reporter gene in the COS-1 and/or MDCK cells can be altered drastically by modifying the concentrations of the recombinant plasmids transfected in the COS-1 cells. Thus, it was observed that overexpression of NS2 reduced CAT expression in COS-1 cells, whereas overexpression of M2 and NS1 proteins dramatically decreased transmission of the CAT RNA to the MDCK cultures. These results are discussed with reference to the roles of these proteins during virus replication. From these experiments, a ratio of transfected plasmids was found that increased the efficiency of the previously described system by 50-100-fold. Under these optimized conditions, it was demonstrated that VLPs can be formed in the absence of neuraminidase expression and that these VLPs remained aggregated to each other and to cell membranes. Moreover, it is shown that CAT RNA transmission was dependent on specific interactions of the ribonucleoprotein complex with other viral structural polypeptides. These data demonstrate the usefulness of this encapsidation-packaging system for the study of different aspects of the influenza virus life-cycle.

Animals↗

Mutational analysis of influenza A virus nucleoprotein: identification of mutations that affect RNA replication.

The influenza A virus nucleoprotein (NP) is a multifunctional polypeptide which plays a pivotal role in virus replication. To get information on the domains and specific residues involved in the different NP activities, we describe here the preparation and characterization of 20 influenza A virus mutant NPs. The mutations, mostly single-amino-acid substitutions, were introduced in a cDNA copy of the A/Victoria/3/75 NP gene and, in most cases, affected residues located in regions that were highly conserved across the NPs of influenza A, B, and C viruses. The mutant NPs were characterized (i) in vivo (cell culture) by analyzing their intracellular localization and their functionality in replication, transcription, and expression of model RNA templates; and (ii) in vitro by analyzing their RNA-binding and sedimentation properties. The results obtained allowed us to identify both a mutant protein that accumulated in the cytoplasm and mutations that altered the functionality and/or the oligomerization state of the NP polypeptide. Among the mutations that reduced the NP capability to express chloramphenicol acetyltransferase protein from a model viral RNA (vRNA) template, some displayed a temperature-sensitive phenotype. Interestingly, four mutant NPs, which showed a reduced functionality in synthesizing cRNA molecules from a vRNA template, were fully competent to reconstitute complementary ribonucleoproteins (cRNPs) capable of synthesizing vRNAs, which in turn yielded mRNA molecules. Based on the phenotype of these mutants and on previously published observations, it is proposed that these mutant NPs have a reduced capability to interact with the polymerase complex and that this NP-polymerase interaction is responsible for making vRNPs switch from mRNA to cRNA synthesis.

Amino Acid Sequence↗

In vivo reconstitution of active Thogoto virus polymerase: assays for the compatibility with other orthomyxovirus core proteins and template RNAs.

Tick-borne Thogoto virus (THOV), the prototype of a new genus in the Orthomyxoviridae family, contains six single-stranded RNA segments of negative polarity. Four of them encode gene products that correspond to the influenza virus PB1, PB2, PA and NP core proteins. Here we describe an in vivo system in which the expression of a THOV model RNA is driven by THOV core proteins synthesized from cloned cDNAs. Our results demonstrated the biological activity of our cloned genes and showed that the three polymerase subunits and the NP are required for gene expression. For comparison, we also used the in vivo reconstituted systems of the influenza A and B viruses. None of the polymerase or NP proteins was active in a heterologous orthomyxovirus core, indicating a high specificity in core assembly and/or function. Interestingly, the THOV polymerase did not recognize the influenza A virus promoter and vice versa.

Cloning, Molecular↗

[Coronary intervention at a general hospital in the State of Piauí, Brazil].

PURPOSE: To evaluate retrospectively success and complication rates of coronary interventions performed in a general hospital with no digital equipment in the State of Piauí with a small annual number of procedures. METHODS: From August/91 to January/98, 122 patients underwent 146 procedures using a fluoroscope with camera and TV monitor of 525 lines. The variables analyzed included the initial success of procedures (residual stenosis less than fifty percent and TIMI III flow) and more serious complications (acute occlusion, infarcts, emergency surgeries and death). The clinical data and the morphological characteristics of the lesions (ACC/AHA Task Force) were also analyzed. RESULTS: Eighty five percent of the patients presented with an acute ischemic syndrome (acute myocardial infarction, post infarction angina, rest angina), 5% were in cardiogenic shock and 88% had type B lesions. The initial success rate was 93% while the more serious complications occurred in 3.5% of the cases. CONCLUSION: In spite of the small annual procedural volume and the use of conventional equipment, the initial success rate in our institution was excellent in a diverse and complex patient population while the complication rates were similar to that of the literature.

Adult↗

The three subunits of the polymerase and the nucleoprotein of influenza B virus are the minimum set of viral proteins required for expression of a model RNA template.

The genes encoding the nucleoprotein, PB1, PB2, and PA proteins of the influenza virus strain B/Panamá/45/90 have been cloned under control of the T7 RNA polymerase promoter of plasmid pGEM-3. Transfection of the recombinant plasmids obtained into mammalian cells, which had been infected with a vaccinia virus encoding the T7 RNA polymerase, resulted in expression of the expected influenza B virus polypeptides. Moreover, it is shown that coexpression of the four recombinant core proteins in COS-1 cells reconstituted a functional polymerase capable of expressing a synthetic influenza B virus-like CAT RNA. By using the influenza B virus recombinant plasmids and a set of pGEM-derived plasmids encoding the homologous core proteins of the influenza A virus A/Victoria/3/75 (I. Mena et al. (1994). J. Gen. Virol. 75, 2109-2114), the capabilities of homo- and heterotypic mixtures of the four core proteins to express synthetic type A and B CAT RNAs were analyzed. Both the influenza A and B virus polymerases were active in expressing, albeit with reduced efficiencies, the heterotypic model CAT RNAs. However, none of all possible heterotypic mixtures of the core proteins reconstituted a functional polymerase. In order to fully characterize the recombinant plasmids obtained, the nucleotide sequences of the cloned genes were determined and compared to sequences of other type B virus isolates. The results obtained from these latter analyses are discussed in terms of the conservation and evolution of the influenza B virus core genes.

Amino Acid Sequence↗

Clinical relevance of about-yearly changes in blood pressure and the environment.

Changes identified by inferential statistics from summer to winter of high to low arterial blood pressures (BP) have been quantified as a large predictable about-yearly BP swing. This condition of a large annual BP amplitude (LABPA) raises concern about hypotension as well as hypertension and raises new questions regarding appropriate guidelines for diagnosis and treatment. Recommendations made in the fall on the basis of data collected in the summer may be totally inadequate in dealing with the patient's condition in the winter. In order to avoid such mistakes, it is imperative to implement a systematic surveillance of BP in the light of current chronobiological limits. Patients with a large circannual BP amplitude are particularly suited for a study of the underlying hormonal mechanisms. The longitudinal monitoring of their BP is also amenable to the study of environmental influences from near and far.

Aldosterone↗

Changes in human blood pressure with season, age and solar cycles: a 26 year record.

The record was studied of a 71-year-old, diurnally active (0700-2200 hours) male psychiatrist (G.N.) who self-measured systolic and diastolic blood pressure (BPS and BPD) mostly but not exclusively on Sunday mornings, from 1969 to 1994. A large about-yearly change was revealed which increased with age and was accompanied by a decreasing trend in the yearly rhythm-adjusted mean (MESOR; P < 0.01). According to conventional criteria that specify only upper limits of acceptability, G.N. was hypertensive in summer and normotensive in other seasons. Since changes in both MESOR and circannual amplitude occurred, a systematic surveillance of BP is the chronobiological recommendation.

Age Factors↗

The circadian rhythm of cortisol in the saliva of young pigs.

Single and population-mean cosinor analyses document a circadian rhythm in salivary cortisol of pigs (p < 0.001). The midline estimated statistic of rhythm, the MESOR (M), is 1.50 +/- 0.07 ng/ml. For the group of 14 pigs studied there was a predictable variation of 64% around this mean in salivary cortisol within a day: the double circadian amplitude, 2A, was 0.96 ng/ml, with a 95% confidence interval ranging from 0.60 to 1.32 ng/ml. A measure of timing, the acrophase, phi, was 10 h 52 min from the phase reference (0030, the middle of the daily dark span of the 24-h light-dark cycle in which the pigs were kept). The 95% confidence interval of phi extended from 9 h 48 min to 12 h 12 min from the reference time. The chronobiologic approach provides indispensable quantitive characteristics as reference standards for future basic or applied work.

Animals↗

A point mutation in the F1 subunit of human respiratory syncytial virus fusion glycoprotein blocks its cell surface transport at an early stage of the exocytic pathway.

Vaccinia virus recombinants expressing either wild-type or mutant forms of human respiratory syncytial (RS) virus (Long strain) fusion (F) glycoprotein were obtained. Proteolytic processing of the precursor, F0, and cell surface transport of the F glycoprotein were unaffected in the recombinants, except in those that contained the replacement Phe --> Ser at position 237 of the F1 subunit. In recombinants containing this mutation, either alone or in combination with others, the traffic of the F molecule was arrested at some intermediate step of its transport to the cell surface and, consequently, the endoproteolytic cleavage of the F0 precursor was inhibited. Immunofluorescence staining of infected cells and endoglycosidase H (Endo-H) sensitivity assays indicated that the arrest occurred before the mid-Golgi compartment. Dimerization and folding of the F protein were also affected by the Phe237 --> Ser substitution. Other amino acid replacements at positions 236 or 237 of the F1 subunit had various effects upon F0 maturation. These results are discussed in terms of the maturation requirements for the RS virus F molecule.

Animals↗