Biothics in Brazil

In this article the authors briefly sketch the nature of Brazilian bioethics. Bioethics emerged in Brazil later than in other Western countries and the 1990's were the most important period for the spread of the discipline in the country. It is in this period that some structural elements of bioethics were established, such as research groups, regulation of Local Research Ethics Committees (Comitês Locais de Ética em Pesquisa – CEP), the creation of the National Commission of Ethics in Research with Human Beings (Comissão Nacional de Ética em Pesquisa com Seres Humanos – CONEP) and the Brazilian Bioethics Society (Sociedade Brasileira de Bioética – SBB). With regard to theoretical work, Brazilian bioethics is clearly an importer of theories from countries central to the studies of bioethics, or, in another words, countries where bioethics first emerged and was established. The most commonly used theory among Brazilian researchers is principlism     

Gas chromatographic assay of cellular fatty acids and alcohols for the identification of Mycobacterium species.

Ten mycobacterial species obtained from 141 cultures isolated from clinical specimens were studied. The cultures were grown on solid medium and then analysed-after saponification, methylation, extraction with organic solvent and washing of the organic phase--by capillary gas-liquid chromatography for fatty acid and secondary alcohol composition. The absence of secondary alcohols was characteristic of M. genavense, M. tuberculosis and the following Mycobacterium species with specific branched-chain fatty acids allowing their direct identification: M. gordonae, M. kansasii and M. marinum. The presence of secondary alcohols was characteristic of M. avium, M. phlei, M. scrofulaceum, M. terrae and M. xenopi. In the case of M. xenopi direct identification was made possible by the presence of a specific alcohol.     

Psychiatric examinations on handcuffed convicts in Brazil: ethical concerns

Psychiatric examinations in official institutions of the Brazilian government include examinations of individual convicts – some of whom are highly dangerous – carried out by court decision. These individuals are taken handcuffed under police escort from penitentiaries to the examination site. In most Brazilian states, medical examiners or experts adopt the basic procedure of asking the police officers to remove the handcuffs from the convict for the examination to be carried out. This article analyzes, from the bioethical standpoint, the behavior of 49 forensic psychiatrists in eight Brazilian states in relation to procedures adopted as protection measures against any possible danger posed by their examinees. In the context of a developing country, where sophisticated protection means are not available, the practice of keeping convicts handcuffed during such examinations is compared to other measures and more advisable alternatives are discussed from the ethical standpoint. Considering the dilemma between showing minimum respect for the autonomy of examinees and ensuring the security of examiners, the need to adopt equitable procedures based on the recognition of the different needs of examiners and examinees is addressed.     

Faecal Contamination (Viral and Bacteria) Detection in Groundwater used for Drinking Purposes in São Paulo,Brazil

This project has been developed to evaluate the possible relationship between the cesspit (pit latrine) in as far as it degrades the quality of underground water. Its importance is due to the fact that in the rural communities in the State of São Paulo (Brazil) this type of cesspit is very common as a means of sewage disposal and these communities use underground water for their supply of drinking water. Rural properties distributed over the rural area in the municipality of São José do Rio Preto were selected. A preliminary study was then set up to determine the social situation and health of the households as well as qualitative evaluations on the type of water supply and sewage disposal of these communities. Campaigns of water sampling then followed and laboratory analyses of water taken from wells were carried out. Parameters were set up to evaluate the potability according to Brazilian legislation (2004) paying attention to microbiologic (coliforms, Crytosporidium sp., and adenovirus). The analyses showed evidence of possible inter-action between the wells and the sewage effluents and drainage in these communities. A PCR reaction to detect adenovirus showed a presence in 53.3% of the samples. The tests for the detection of Cryotosporidium sp all showed a negative result.     

Isolation and characterization of lymphatic microvascular endothelial cells from human tonsils

Human lymphatic endothelial cells (LECs) have isolated prevalently from human derma and tumors. As specialized lymphatic organs within the oropharynx, palatine tonsils are easily obtained and rich in lymphatic venules. Using a two-step purification method based on the sorting of endothelial cells with Ulex Europaeus Agglutinin 1 (UEA-1)-coated beads, followed by purification with monoclonal antibody D2-40, we successfully purified LECs from human palatine tonsils. The LECs were expanded on flasks coated with collagen type 1 and fibronectin for up to 8-10 passages and then analyzed for phenotypic and functional properties. Cultured cells retained the phenotypic pattern of the lymphatic endothelium of palatine tonsils and expressed functional VEGFR-3 molecules. In fact, stimulation with VEGFR-3 ligand, the vascular endothelium grow factor C, induced a marked increase in cell proliferation. Similarly to blood endothelial cells (BECs), LECs were able to form tube-like structure when seeded in Cultrex basement membrane extract. Comparative studies performed on LECs derived from palatine tonsils and iliac lymphatic vessels (ILVs), obtained with the same procedures, showed substantial discrepancies in the expression of various lymphatic markers. This points to the existence of micro- and macrovessel-derived LECs with different phenotypes, possibly involving different biological activities and functions. Palatine tonsil- and ILV-derived LECs may, therefore, represent new models for investigating function and biochemical properties of these lymphatic endothelia.     

Replication-deficient mutant Herpes Simplex Virus-1 targets professional antigen presenting cells and induces efficient CD4+ T helper responses

Both neutralizing antibodies and cytotoxic T-cells are necessary to control a viral infection. However, vigorous T helper responses are essential for their elicitation and maintenance. Here we show that a recombinant replication-deficient Herpes Simplex Virus (HSV)-1 vector encoding the Human Immunodeficiency Virus (HIV)-1 matrix protein p17 (T0-p17) was capable of infecting professional antigen presenting cells (APCs) in vitro and in vivo. The injection of T0-p17 in the mouse dermis generated a strong p17-specific CD4+ T helper response preceding both p17-specific humoral and effector T cell responses. Moreover, we show that T0-p17 infection did not interfere with the endogenous processing of the transgene encoded antigen, since infected APCs were able to evoke a strong recall response in vitro. Our results demonstrate that replication-deficient HSV vectors can be appealing candidates for the development of vaccines able to trigger T helper responses.     

Intervention bioethics: a proposal for peripheral countries in a context of power and injustice

The bioethics of the so-called 'peripheral countries' must preferably be concerned with persistent situations, that is, with those problems that are still happening, but should not happen anymore in the 21st century. Resulting conflicts cannot be exclusively analysed based on ethical (or bioethical) theories derived from 'central countries.' The authors warn of the growing lack of political analysis of moral conflicts and of human indignation. The indiscriminate utilisation of the bioethics justification as a neutral methodological tool softens and even cancels out the seriousness of several problems, even those that might result in the most profound social distortions. The current study takes as a theoretical reference the fact that natural resources (which affect us all) are relevant. Based on these premises, and on the concept that equity means 'treating unevenly the unequal', a proposal of a hard bioethics (or intervention bioethics) is introduced, in defence of the historical insights and rights of economically and socially excluded populations that are separated from the international developmental process.     

Heterogeneous expression of toll-like receptors in lymphatic endothelial cells derived from different tissues

As lymphatic endothelial cells (LECs) express different lymphatic and vascular markers depending on the organ they are derived from, we analysed whether they also show a heterogeneity of response against pathogens. To this end we analysed, for the presence of mRNA encoding for all human toll-like receptor (TLR), LECs isolated from lymph nodes and thymuses. RNA for TLR1-6 and 9 was identified in thymus-derived cells, whereas cells derived from lymph nodes contained mRNA for TLR1-4, 6 and 9, but failed to express mRNA specific for TLR5. The differential expression of TLRs was confirmed by the phosphorylation of nuclear factor-κB p65 only when the two types of LECs were incubated with the appropriate TLR agonists. The stimulation with specific agonists gives rise to a heterogeneous pattern of cytokine and chemokine secretion: thymus-derived LECs produced preferentially interleukin-6, interferon-inducible protein (IP)-10 and tumour necrosis factor-α, whereas cells prepared from lymph nodes mainly released interleukin-8, monocyte chemotactic protein-1, RANTES and (IP)-10. Finally, cells purified from lymph nodes expressed a higher level of intercellular adhesion molecule-1 than did cells prepared from the thymus when stimulated with several TLR agonists. The expression of a large set of TLRs and the responsiveness to specific agonists suggest that LECs are able to respond to pathogens, and the observed differences reflect specialized functions, redundancy and/or roles of LECs of different origin.     

Development of horse polyclonal antiserum inhibiting all in vitro biological functions of human IFN-gamma

Following a standard immunization protocol with recombinant human interferon-gamma (IFN-gamma), a horse polyclonal antiserum was obtained and evaluated for its ability to interfere with multiple IFN-gamma activities in vitro. Data obtained show that polyclonal horse antiserum neutralizes the antiproliferative activity of IFN-gamma, inhibits the binding of IFN-gamma to cellular receptors, and can up-regulate HLA-DR antigen expression and interfere with its antiviral activity. The broad neutralizing capacity of horse polyclonal antiserum has been assessed on cell lines which differ as to origin and sensitivity to IFN-gamma. Moreover, we observed that this antiserum could inhibit the binding of radiolabeled IFN-gamma to its cellular receptor, its subsequent internalization into the target cell, and its antiviral activity. As it is able to inhibit all the biological activities of IFN-gamma, this antiserum might provide new therapeutic approaches to diseases with evidence of activated cell-mediated immunity.