Study of the evaluation criteria for teeth injuries by judges, experts in forensic departments and forensic dentistry experts in accordance to the penal law

Orientadores: Eduardo Daruge, Eduardo Daruge JúniorDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de PiracicabaResumo: Com o passar dos anos tem se constatado um aumento da violência, e como conseqüência, do número de exames de corpo de delito envolvendo a face e cavidade bucal. Mesmo existindo coeficientes dos índices estético, mastigatório e fonético, nota-se uma evidente falta de padronização na avaliação e enquadramento das lesões dentárias de acordo com o artigo 129 do Código Penal. Devido a este fato o presente estudo avaliou a maneira como os juízes, peritos dos Institutos Médico Legais (IMLs), entre eles médicos e cirurgiões dentistas, e especialistas em Odontologia Legal, tipificam as lesões dentárias; assim como comparou as convergências e divergências das opiniões desses profissionais e discutiu os aspectos éticos e legais pertinentes ao tema. Nesse objetivo, foi confeccionado um questionário com questões estruturadas, que foi entregue aos voluntários, juntamente com duas cópias do termo de consentimento livre e esclarecido (TCLE). O projeto da presente pesquisa foi aprovado pelo Comitê de Ética em Pesquisa sob o protocolo nº 076/2009. A amostra foi de 82 profissionais, atuantes nos Estados de São Paulo, Rio de Janeiro e Mato Grosso, que qualificaram supostas lesões dentárias de acordo com o artigo 129 do Código Penal. Conclui-se que apesar da existência de uma tendência à convergência nas opiniões dos profissionais, quando observadas as porcentagens maiores, existe uma oscilação na interpretação das questões, fato que dificulta a aplicação de um critério único. Pelo contrário, se existissem parâmetros para tal fim, diminuiriam as possibilidades de variações na forma de interpretação entre profissionais diretamente ligados no processo, no tocante aos danos sofridos pela vítimaAbstract: Through the years, it has been noticed an increase of violence and forensic examination cases related to the face and oral cavity. Although there are aesthetic, phonetic and masticatory indexes, there is an obvious lack of criteria for the assessment and classification of dental injuries in accordance to 129th Article of the Penal Code. Due to this fact, this study analyzed how judges, medical and dental forensic experts, and specialists in forensic dentistry typify the dental injuries, as well as checked the convergence and divergence of opinions of these professionals and discussed the ethical and legal issues referred to this topic. In this goal, it was made a questionnaire with structured questions, which was delivered to the volunteers, along two copies of the informed consent (IC). The project of this research was approved by the Research Ethics Committee (protocol nº 076/2009). The sample consisted of 82 professionals from the States of São Paulo, Rio de Janeiro and Mato Grosso, that qualified supposed dental injuries in accordance with 129th Article of the Penal Code. It was concluded that despite of the existence of a convergent tendency in the points of view of professionals, while observing the highest percentages, there are discrepancies in the interpretation of the questions, that difficult the application of a single criterion. Although, if there was a parameter for this purpose, it would minimize the possibility of different interpretations among the professionals directly involved in the process, when related to the damages suffered by the victimMestradoOdontologia Legal e DeontologiaMestre em Biologia Buco-Denta

Identification and functional characterization of a highly divergent N-acetylglucosaminyltransferase I (TbGnTI) in <em>Trypanosoma brucei</em>

Trypanosoma brucei expresses a diverse repertoire of N-glycans, ranging from oligomannose and paucimannose structures to exceptionally large complex N-glycans. Despite the presence of the latter, no obvious homologues of known β1–4-galactosyltransferase or β1–2- or β1–6-N-acetylglucosaminyltransferase genes have been found in the parasite genome. However, we previously reported a family of putative UDP-sugar-dependent glycosyltransferases with similarity to the mammalian β1–3-glycosyltransferase family. Here we characterize one of these genes, TbGT11, and show that it encodes a Golgi apparatus resident UDP-GlcNAc:α3-d-mannoside β1–2-N-acetylglucosaminyltransferase I activity (TbGnTI). The bloodstream-form TbGT11 null mutant exhibited significantly modified protein N-glycans but normal growth in vitro and infectivity to rodents. In contrast to multicellular organisms, where the GnTI reaction is essential for biosynthesis of both complex and hybrid N-glycans, T. brucei TbGT11 null mutants expressed atypical “pseudohybrid” glycans, indicating that TbGnTII activity is not dependent on prior TbGnTI action. Using a functional in vitro assay, we showed that TbGnTI transfers UDP-GlcNAc to biantennary Man(3)GlcNAc(2), but not to triantennary Man(5)GlcNAc(2), which is the preferred substrate for metazoan GnTIs. Sequence alignment reveals that the T. brucei enzyme is far removed from the metazoan GnTI family and suggests that the parasite has adapted the β3-glycosyltransferase family to catalyze β1–2 linkages

An essential, kinetoplastid-specific GDP-Fuc:β-D-Gal α-1,2-fucosyltransferase is located in the mitochondrion of <i>Trypanosoma brucei</i>

Fucose is a common component of eukaryotic cell-surface glycoconjugates, generally added by Golgi-resident fucosyltransferases. Whereas fucosylated glycoconjugates are rare in kinetoplastids, the biosynthesis of the nucleotide sugar GDP-Fuc has been shown to be essential in Trypanosoma brucei. Here we show that the single identifiable T. brucei fucosyltransferase (TbFUT1) is a GDP-Fuc: β-D-galactose α-1,2-fucosyltransferase with an apparent preference for a Galβ1,3GlcNAcβ1-O-R acceptor motif. Conditional null mutants of TbFUT1 demonstrated that it is essential for both the mammalian-infective bloodstream form and the insect vector-dwelling procyclic form. Unexpectedly, TbFUT1 was localized in the mitochondrion of T. brucei and found to be required for mitochondrial function in bloodstream form trypanosomes. Finally, the TbFUT1 gene was able to complement a Leishmania major mutant lacking the homologous fucosyltransferase gene (Guo et al., 2021). Together these results suggest that kinetoplastids possess an unusual, conserved and essential mitochondrial fucosyltransferase activity that may have therapeutic potential across trypanosomatids

The GPI-Phospholipase C of Trypanosoma brucei Is Nonessential But Influences Parasitemia in Mice

In the mammalian host, the cell surface of Trypanosoma brucei is protected by a variant surface glycoprotein that is anchored in the plasma membrane through covalent attachment of the COOH terminus to a glycosylphosphatidylinositol. The trypanosome also contains a phospholipase C (GPI-PLC) that cleaves this anchor and could thus potentially enable the trypanosome to shed the surface coat of VSG. Indeed, release of the surface VSG can be observed within a few minutes on lysis of trypanosomes in vitro. To investigate whether the ability to cleave the membrane anchor of the VSG is an essential function of the enzyme in vivo, a GPI-PLC null mutant trypanosome has been generated by targeted gene deletion. The mutant trypanosomes are fully viable; they can go through an entire life cycle and maintain a persistent infection in mice. Thus the GPI-PLC is not an essential activity and is not necessary for antigenic variation. However, mice infected with the mutant trypanosomes have a reduced parasitemia and survive longer than those infected with control trypanosomes. This phenotype is partially alleviated when the null mutant is modified to express low levels of GPI-PLC

Genetic Metabolic Complementation establishes a requirement for GDP-fucose in <i>Leishmania</i>

To survive in its sand fly vector, the trypanosomatid protozoan parasite Leishmania first attaches to the midgut to avoid excretion, but eventually it must detach for transmission by the next bite. In Leishmania major strain Friedlin, this is controlled by modifications of the stage-specific adhesin lipophosphoglycan (LPG). During differentiation to infective metacyclics, d-arabinopyranose (d-Arap) caps the LPG side-chain galactose residues, blocking interaction with the midgut lectin PpGalec, thereby leading to parasite detachment and transmission. Previously, we characterized two closely related L. major genes (FKP40 and AFKP80) encoding bifunctional proteins with kinase/pyrophosphorylase activities required for salvage and conversion of l-fucose and/or d-Arap into the nucleotide-sugar substrates required by glycosyltransferases. Whereas only AFKP80 yielded GDP-d-Arap from exogenous d-Arap, both proteins were able to salvage l-fucose to GDP-fucose. We now show that Δafkp80− null mutants ablated d-Arap modifications of LPG as predicted, whereas Δfkp40− null mutants resembled wild type (WT). Fucoconjugates had not been reported previously in L. major, but unexpectedly, we were unable to generate fkp40−/afkp80− double mutants, unless one of the A/FKPs was expressed ectopically. To test whether GDP-fucose itself was essential for Leishmania viability, we employed “genetic metabolite complementation.” First, the trypanosome de novo pathway enzymes GDP-mannose dehydratase (GMD) and GDP-fucose synthetase (GMER) were expressed ectopically; from these cells, the Δfkp40−/Δafkp80− double mutant was now readily obtained. As expected, the Δfkp40−/Δafkp80−/+TbGMD-GMER line lacked the capacity to generate GDP-Arap, while synthesizing abundant GDP-fucose. These results establish a requirement for GDP-fucose for L. major viability and predict the existence of an essential fucoconjugate(s)

A gene of the β3-glycosyltransferase family encodes <i>N</i>-acetylglucosaminyltransferase II function in <i>Trypanosoma brucei</i>

The bloodstream form of the human pathogen Trypanosoma brucei expresses oligomannose, paucimannose and complex N-linked glycans, including some exceptionally large poly-N-acetyllactosamine-containing structures. Despite the presence of complex N-glycans in this organism, no homologues of the canonical N-acetylglucosaminyltransferase I or II genes can be found in the T. brucei genome. These genes encode the activities that initiate the elaboration of the Manalpha1-3 and Manalpha1-6 arms, respectively, of the conserved trimannosyl-N-acetylchitobiosyl core of N-linked glycans. Previously, we identified a highly divergent T. brucei N-acetylglucosaminyltransferase I (TbGnTI) among a set of putative T. brucei glycosyltransferase genes belonging to the beta3-glycosyltransferase superfamily (1). Here, we demonstrate that TbGT15, another member of the same beta3-glycosyltransferase family, encodes an equally divergent N-acetylglucosaminyltransferase II (TbGnTII) activity. In contrast to multicellular organisms, where GnTII activity is essential, TbGnTII null mutants of T. brucei grow in culture and are still infectious to animals. Characterization of the large poly-N-acetyllactosamine containing N-glycans of the TbGnTII null mutants by methylation linkage analysis suggests that, in wild-type parasites, the Manalpha1-6 arm of the conserved trimannosyl core may carry predominantly linear poly-N-acetyllactosamine chains whereas the Manalpha1-3 arm may carry predominantly branched poly-N-acetyllactosamine chains. These results provide further detail on the structure and biosynthesis of complex N-glycans in an important human pathogen and provide a second example of the adaptation by trypanosomes of beta3-glycosyltransferase family members to catalyze beta1-2 glycosidic linkages

Thermodynamic qualification of knitted spacer fabrics for use as insulation box insert in the context of refrigerated transport containers in the logistics sector

Temperature-sensitive products such as refrigerated and frozen goods pose particular challenges for logistics. Against the background of the mobility shift towards electric vehicles and the current challenges of temperature-stable transport in the field of pharmaceutical, esp. vaccine logistics in the context of the SARS-CoV-2 pandemic, new, energy-efficient vehicle equipment is needed to maintain cold chains. Known refrigeration concepts are designed to cool the entire cargo hold. In addition, the goods cannot be removed from the vehicle while maintaining the cold chain. An insulating effect of containers is typically achieved by using foamed polystyrene (Styrofoam). On the one hand, these structures have a very good insulating effect, but on the other hand, they cannot be reduced in volume during recirculation and are problematic with regard to recycling. The aim of the research presented here is therefore to develop a knitted box that is designed as a volume-reducible, rigid but foldable box. This can be used as a supplement to existing transport container systems and therefore can be inserted in the transport container. The knitted box performs as insulation when the transported goods are actively cooled inside the box, which is more sustainable and flexible than recent insulation solutions. Knitted fabrics, especially spacer fabrics, have advantageous thermo-physical properties for this application due to their structural design. In the course of a research project, various spacer fabrics were tested for their thermo-physical suitability as insulation materials. It was found that knitted predetermined folding lines represent an insulation gap. Based on this, a new structure was developed which, due to its structural design, compensates for cold or thermal bridges at vertices and edges of the box. The results show that the knitted corrugated structure insulates better than the knitted spacer fabrics with predetermined folding lines. A thermal imaging camera was used to identify critical points for heat transfer