Quality evaluation of DNA obtained from stored human saliva and its applicability to identification in Forensic Dentistry

PURPOSE: This study evaluated the quality of DNA obtained from stored human saliva and its applicability to human identification. METHODS: The saliva samples of 20 subjects, collected in the form of saliva in natura and from mouth swabs and stored at -20ºC, were analyzed. After 7 days, the DNA was extracted from the 40 saliva samples and subjected to PCR and electrophoresis. After 180 days, the technique was repeated with the 20 swab samples. RESULTS: The first-stage results indicated that DNA was successfully extracted in 97.5% of reactions, 95% of saliva in natura and 100% of swab saliva samples, with no statistically significant difference between the forms of saliva. In the second phase, the result was positive for all 20 analyzed samples (100%). Subsequently, in order to analyze the quality of the DNA obtained from human saliva, the SIX3-2 gene was tested on the 20 mouth swab samples, and the PCR products were digested using the MbO1 restriction enzyme to evaluate polymorphisms in the ADRA-2 gene, with positive results for most samples. CONCLUSION: It was concluded that the quantity and quality of DNA from saliva and the techniques employed are adequate for forensic analysis of DNA.

A utilização de imagens na identificação humana em odontologia legal

Este artigo de revisão sistemática tem por objetivo citar os métodos de identificação humana por meio da radiologia, utilizados em odontologia legal. Para isso, realizou-se revisão de literatura com 19 trabalhos selecionados dentre 45 encontrados, após aplicação de critérios de inclusão. Há diversas técnicas radiológicas que podem ser utilizadas para auxiliar na identificação humana, tanto individual como geral, incluindo a determinação do gênero, do grupo étnico e, principalmente, da idade. A análise de radiografias e tomografias ante-mortem e post-mortem tornou-se uma ferramenta fundamental nos processos de identificação em odontologia legal, principalmente com o refinamento das técnicas adquiridas com o avanço da própria radio-logia e com a incorporação da informática. Conclui-se que a partir do conhecimento adequado dos métodos disponíveis, o profissional em odontologia legal pode optar pelo método que melhor preencha as características necessárias para o sucesso da identificação, tendo cuidado na aplicação correta da técnica e na interpretação precisa das informações obtidas.

Interleukin-22 forms dimers that are recognized by two interleukin-22R1 receptor chains

Interleukin-22 (IL-22) is a class 2 cytokine whose primary structure is similar to that of interleukin 10 (IL-10) and interferon-gamma (IFN-gamma). IL-22 induction during acute phase immune response indicates its involvement in mechanisms of inflammation. Structurally different from IL-10 and a number of other members of IL-10 family, which form intertwined inseparable V-shaped dimers of two identical polypeptide chains, a single polypeptide chain of IL-22 folds on itself in a relatively globular structure. Here we present evidence, based on native gel electrophoresis, glutaraldehyde cross-linking, dynamic light scattering, and small angle x-ray scattering experiments, that human IL-22 forms dimers and tetramers in solution under protein concentrations assessable by these experiments. Unexpectedly, low-resolution molecular shape of IL-22 dimers is strikingly similar to that of IL-10 and other intertwined cytokine dimeric forms. Furthermore, we determine an ab initio molecular shape of the IL-22/IL-22R1 complex which reveals the V-shaped IL-22 dimer interacting with two cognate IL-22R1 molecules. Based on this collective evidence, we argue that dimerization might be a common mechanism of all class 2 cytokines for the molecular recognition with their respective membrane receptor. We also speculate that the IL-22 tetramer formation could represent a way to store the cytokine in nonactive form at high concentrations that could be readily converted into functionally active monomers and dimers upon interaction with the cognate cellular receptors.

Genotoxic potential generated by biomass burning in the Brazilian Legal Amazon by Tradescantia micronucleus bioassay: a toxicity assessment study

Background: The Brazilian Amazon has suffered impacts from non-sustainable economic development, especially owing to the expansion of agricultural commodities into forest areas. The Tangara da Serra region, located in the southern of the Legal Amazon, is characterized by non-mechanized sugar cane production. In addition, it lies on the dispersion path of the pollution plume generated by biomass burning. The aim of this study was to assess the genotoxic potential of the atmosphere in the Tangara da Serra region, using Tradescantia pallida as in situ bioindicator. Methods: The study was conducted during the dry and rainy seasons, where the plants were exposed to two types of exposure, active and passive. Results: The results showed that in all the sampling seasons, irrespective of exposure type, there was an increase in micronucleus frequency, compared to control and that it was statistically significant in the dry season. A strong and significant relationship was also observed between the increase in micronucleus incidence and the rise in fine particulate matter, and hospital morbidity from respiratory diseases in children. Conclusions: Based on the results, we demonstrated that pollutants generated by biomass burning in the Brazilian Amazon can induce genetic damage in test plants that was more prominent during dry season, and correlated with the level of particulates and elevated respiratory morbidity.

Street children and their relationship with the police

Background: This paper explores and analyses the experiences of school-age street children. It specifically addresses the relationship of the street children who live on the streets of Sao Paulo (a large Brazilian metropolis), in relation to their experiences, with the policemen. Methods: The paper is a secondary analysis of date previously collected in 1999. The data were collected through individual semi-structured interviews, with 14 school-age children frequenting two city public refuges, with their legal guardians` consent. The text from transcribed interviews was organized according to the validity norms of `thematic analysis`, a technique of contents analysis method. The decomposing and reconstructing process of that analysis gave rise to thematic categories (among which `the police category`) that represented the reconstruction of the difficulties faced by the children in their development. Results and discussion: The children portrayed the police as an enemy, a fearful figure and one of the most agonizing street experiences. Rarely did the police have a positive image to them. According to the children, police violence occurs in three forms: through systematic police persecution in an effort to remove the children from the streets against their will; actions that had the deliberate intent to humiliate them with verbal or physical aggression; and through alleged sexual abuse, revealed by the children in a veiled manner. The authority that is supposedly intended to protect them is portrayed as one of the most feared social agents. Conclusion: The reported hostile behaviour of the policemen shows the state of vulnerability of those children living on the street. This situation must be focused like a health problem because it causes injury to development of children. Nurses can help them through organizing assistance to children in situation of personal and social risk in the school nursing and health institution.

Synthesis of Cyclic Guanidine Intermediates of Anatoxin-a(s) in Both Racemic and Enantiomerically Pure Forms

The alkyl chain of anatoxin-a(s) (cyclic guanidines), which can be used as an intermediate in the total synthesis of anatoxin-a(s), was synthesized in both racemic and enantiomerically pure forms. These enantiomerically pure cyclic compounds can be used as chiral inductors in some reactions. The two racemic routes disclosed herein have the advantages of high overall yield and mild reaction conditions. Both routes proceed through an intermediate 2,3-diaminoacid - an important synthetic scaffold - with good yields. Furthermore, the N,N-dimethyl-2(tosylimino)imidazolidine-4-carboxamide might be obtained from 2-(tosylimino)imidazolidine-4-carboxylic acid followed by selective reduction of the carbonyl functionality. All synthesized compounds were analyzed by mass spectrometry and (1)H NMR and (13)C NMR spectroscopy.

Monocyte-derived dendritic cells from patients with severe forms of chromoblastomycosis induce CD4(+) T cell activation in vitro

Dendritic cells (DCs) have been described as initiators and modulators of the immune response. Recently we have shown a predominant production of interleukin-10 cytokine, low levels of interferon-gamma and inefficient T cell proliferation in patients with severe forms of chromoblastomycosis. Chromoblastomycosis starts with subcutaneous inoculation of Fonsecaea pedrosoi into tissue where DCs are the first line of defence against this microorganism. In the present study, the interaction of F. pedrosoi and DCs obtained from patients with chromoblastomycosis was investigated. Our results showed that DCs from patients exhibited an increased expression of human leucocyte antigen D-related (HLA-DR) and co-stimulatory molecules. In the presence of conidia, the expression of HLA-DR and CD86 was up-regulated by DCs from patients and controls. Finally, we demonstrate the reversal of antigen-specific anergy and a T helper type 1 response mediated by DCs incubated with F. pedrosoi conidea.

Standing wave bed forms at Plage de la Grande Roche, le Verdelet, Pléneuf-Val-André, Cote d’Armor (France)

Coastal Photograph by Hubert Chanson This photograph of standing wave bed forms was taken at very low tide. The tidal range was 10 m. The bed forms were located on the island of Le Verdelet, in a channel between Le Grande Jaune and Le Verdelet. It is likely that these standing wave bed forms were formed during transcritical shallow water flows at the end of ebb tide. The author’s watch is in the foreground for scale. (Coastal Photograph by Hubert Chanson, Division of Civil Engineering, the University of Queensland, Brisbane, Queensland 4072, Australia.)

Cost-benefit analysis of alternative forms of hedgerow intercropping in the Philippine uplands

Considerable resources have been expended promoting hedgerow intercropping with shrub legumes to farmers in the Philippine uplands. Despite the resources committed to research and extension, persistent adoption by farmers has been limited to low cost versions of the technology including natural vegetation and grass strips. In this paper, cost-benefit analysis is used to compare the economic returns from traditional open-field maize farming with returns from intercropping maize between leguminous shrub hedgerows, natural vegetation strips and grass strips. An erosion/productivity model, Soil Changes Under Agroforestry, was used to predict the effect of erosion on maize yields. Key informant surveys with experienced maize farmers were used to derive production budgets for the alternative farming methods. The economic incentives revealed by the cost-benefit analysis help to explain the adoption of maize farming methods in the Philippine uplands. Open-field farming without hedgerows has been by far the most popular method of maize production, often with two or more fields cropped in rotation. There is little persistent adoption of hedgerow intercropping with shrub legumes because sustained maize yields are not realised rapidly enough to compensate farmers for establishment and maintenance costs. Natural vegetation and grass strips are more attractive to farmers because of lower establishment costs, and provide intermediate steps to adoption. Rural finance, commodity pricing and agrarian reform policies influence the incentives for maize farmers in the Philippine uplands to adopt and maintain hedgerow intercropping.

Familial forms broaden the horizons for primary aldosteronism

The identification of familial forms of primary aldosteronism (PAL) has led to its detection in relatives of affected patients not suspected previously of having PAL. Many ave normokalemic and some ave even normotensive. This broadens the spectrum of PAL, permitting the study of its evolution and of intervention with specific therapy when hypertension develops. The genetic basis of one form involves steroid biosynthetic enzymes and the other form predisposes to hyperplasia and benign neoplasia.

Structures of free and complexed forms of Escherichia coli xanthine-guanine phosphoribosyltransferase

Structures of free, substrate-bound and product-bound forms of Escherichia coli xanthine-guanine phosphoribosyltransferase (XGPRT) have been determined by X-ray crystallography. These are compared with the previously determined structure of magnesium and sulphate-bound XPRT. The structure of free XGPRT at 2.25 Angstrom resolution confirms the flexibility of residues in and around a mobile loop identified in other PRTases and shows that the cis-peptide conformation of Arg37 at the active site is maintained in the absence of bound ligands. The structures of XGPRT complexed with the purine base substrates guanine or xanthine in combination with cPRib-PP, an analog of the second substrate PRib-PP, have been solved to 2.0 Angstrom resolution. In these two structures the disordered phosphate-binding loop of uncomplexed XGPRT becomes ordered through interactions with the 5'-phosphate group of cPRib-PP. The cyclopentane ring of cPRib-PP has the C3 exo pucker conformation, stabilised by the cPRib-PP-bound Mg2+. The purine base specificity of XGPRT appears to be due to water-mediated interactions between the 2-exocyclic groups of guanine or xanthine and side-chains of Glu136 and Asp140, as well as the main-chain oxygen atom of Ile135. Asp92, together with Lys115, could help stabilise the N7-protonated tautomer of the incoming base and could act as a general base to remove the proton from N7 .when the nucleotide product is formed. The 2.6 Angstrom resolution structure of XGPRT complexed with product GMP is similar to the substrate-bound complexes. However, the ribose ring of GMP is rotated by similar to 24 degrees compared with the equivalent ring in cPRib-PP. This rotation results in the loss of all interactions between the ribosyl group and the enzyme in the product complex. (C) 1998 Academic Press.

Availability of two forms of dissolved nitrogen to the coral Pocillopora damicornis and its symbiotic zooxanthellae

The relative contribution of dissolved nitrogen (ammonium and dissolved free amino acids DFAAs) to the nitrogen budget of the reef-building coral Pocillopora damicornis was assessed for colonies growing on control and ammonium-enriched reefs at One Tree Island (southern Great Barrier Reef) during the ENCORE (Enrichment of Nutrient on Coral Reef; 1993 to 1996) project. P. damicornis acquired ammonium at rates of between 5.1 and 91.8 nmol N cm(-2) h(-1) which were not affected by nutrient treatment except in the case of one morph. In this case, uptake rates decreased from 80.5 to 42.8 nmol cm(-2) h(-1) (P < 0.05) on exposure to elevated ammonium over 12 mo. The presence or absence of light during measurement did not influence the uptake of ammonium ions. Nitrogen budgets revealed that the uptake of ammonium from concentrations of 0.11 to 0.13 mu M could completely satisfy the demand of growing P. damicornis for new nitrogen. P. damicornis also took up DFAAs at rates ranging from 4.9 to 9.8 nmol N cm(-2) h(-1). These rates were higher in the dark than in the light (9.0 vs 5.1 nmol m(-2) h(-1), P < 0.001). Uptake rates were highest for the amino acids serine, arginine and alanine, and lowest for tyrosine. DFAA concentrations within the ENCORE microatolls that received ammonium were undetectable, whereas they ranged up to 100 nM within the control microatolls. The contribution of DFAAs to the nitrogen budget of P. damicornis constituted only a small fraction of the nitrogen potentially contributed by ammonium under field conditions. Even at the highest field concentrations measured during this study, DFAAs could contribute only similar or equal to 11.3% of the nitrogen demand of P. damicornis. This contribution, however, may be an important source of nitrogen when other sources such as ammonium are scarce or during periods when high concentrations of DFAAs become sporadically available (e.g. cell breakage during fish-grazing).

Purification,characterization and crystallization in two crystal forms of bovine cyclophilin 40

The purification and crystallization of two different crystal forms of the two-domain protein bovine cyclophilin 40 is reported. Tetragonal crystals grown in methyl pentanediol belong to space group P4(2)22 with unit-cell parameters a = 94.5, c = 118.3 Angstrom. Long thin needles grown from PEG belong to space group C2 with unit-cell parameters a = 125.71, b = 47.3, c = 74.6 Angstrom, beta = 93.90 degrees. The N-terminal 170 amino acids have significant homology with the well characterized human cyclophilin A. The C-terminal domain is largely made up of three copies of the tetratricopeptide repeat motif thought to be involved in mediating protein-protein interactions. Cyclophilins are frequently found as domains in larger multidomain proteins. To date, only X-ray structures of single-domain cyclophilins have been reported, and this work provides the first example of the purification and crystallization of a larger protein containing a cyclophilin domain.