Síntese de moléculas contendo a unidade de piperazina como potenciais agentes antipsicóticos

Tese de Doutoramente em Ciências (área de especialização em Química).

Análise descritiva quantitativa do palmito de pupunheira

Este estudo foi conduzido para descrever o perfil sensorial do palmito de pupunha. A análise descritiva quantitativa (ADQ) foi utilizada para descrever os atributos sensoriais relacionados à aparência, aroma, e textura de três marcas comerciais de palmito de pupunha. As amostras foram avaliadas por uma equipe selecionada com onze provadores e foram definidos nove atributos sensoriais: cor amarela, aparência uniforme e úmida, aroma e sabor não característicos, sabor acido, residual acido e textura macia. Houve diferenças significativas em seis atributos entre as três marcas testadas. Os provadores descreveram os palmitos de pupunha como: cor amarela clara, aparência uniforme e úmida, aroma e sabor não característicos e sabor ácido com valores intermediários, residual acido e amargor final com valores de pouco e textura muito macia a uma textura intermediária.

Photolabile protection for amino acids: studies on the release from novel benzoquinolone cages

The synthesis of a novel fused nitrogen heterocycle, benzoquinolone, for evaluation as a photocleavable protecting group is described for the first time, by coupling to model amino acids (alanine, phenylalanine and glutamic acid). Conversion of the phenylalanine ester conjugate to the thionated derivative was accomplished by reaction with Lawesson’s reagent. Photocleavage studies of the carbonyl and thiocarbonyl benzoquinolone conjugates in various solvents and at different wavelengths (300, 350 and 419 nm) showed that the most interesting result was obtained at 419 nm for the thioconjugate, revealing that the presence of the thiocarbonyl group clearly improved the photolysis rates, giving practicable irradiations times for the release of the amino acids (less than 1 minute).

Aminobenzocoumarinylmehtyl esters as photoactive precursors for the release of butyric acid

The evaluation of the photorelease of a carboxylic acid drug, using butyric acid as a representative model, was carried out by using 7-amino-4-chloromethyl-2-oxo-2Hnaphtho[1,2-b] pyran, an aminobenzocoumarin, and its mono- and di-methylated or ethylated derivatives. This study was intended to improve the release of butyric acid from benzocoumarins by the addition of an amino group to the heterocycle by applying the knowledge of second-generation coumarinylmethyl-based photoremovable protecting groups. Photolysis studies were performed on the resultant ester cages by irradiation in a photochemical reactor at 254, 300, 350 and 419 nm, using methanol/HEPES buffer 80:20 solutions as solvent. The data obtained showed that these new fluorescent aminobenzocoumarins are superior to all the previously tested benzocoumarins with the same or different ring fusions. As well as the photolysis, the photophysics of the compounds were characterised by both steady state and time-resolved methods.

Photoactivation of butyric acid from 6-aminobenzocoumarin cages

A new benzocoumarin bearing an amino group is proposed as a photocleavable protecting group for carboxylic acids. The novel heterocycle, 6-amino-4-chloromethyl-2-oxo-2H-naphtho[1,2-b]pyran was used in the preparation of ester conjugates of butyric acid, and of the corresponding mono- and di-methylated or ethylated derivatives. The photolability of the ester conjugates was studied by irradiation at selected wavelengths in methanol/HEPES buffer (80:20) solutions, and the release of butyric acid was followed with HPLC/UV and 1H NMR monitoring. Release of the carboxylic acid was faster for the monoalkylated derivatives (approximately within 20 min), at the longer wavelengths of irradiation (350 and 419 nm). The photophysics of the heterocyclic conjugates was also evaluated by both steady state and time-resolved methods.

Libertação fotocontrolada de moléculas bioativas a partir de conjugados de cumarinas

Dissertação de mestrado em Química Medicinal

Sonolência diurna excessiva em pré-vestibulandos

OBJETIVO: O sono é um fenômeno que interfere nos aspectos cognitivos. O objetivo deste estudo foi avaliar a prevalência da sonolência diurna excessiva (SDE) em pré-vestibulandos de Campo Grande, MS. MÉTODOS: Foram entrevistados 378 alunos com a escala de sonolência Epworth (ESE). As variáveis foram: sexo, uso esporádico de bebidas alcoólicas e fumo, relato de sinais e sintomas depressivos, renda familiar total dos membros do lar e idade. Foram empregados os testes qui-quadrado e de análise de variância. RESULTADOS: Em relação ao gênero, 50,3% eram homens e 49,7% mulheres; 39,2% ingeriam álcool; 6,6% fumavam e 33% já tinham tido depressão na vida. Entre os alunos, 55,8% tinham SDE, 5,3% eram indicativos de ter distúrbio respiratório ou síndrome da apnéia do sono. Foram detectadas associações entre as variáveis consumo de álcool e tabagismo, em relação à ESE. CONCLUSÕES: Foi alta a prevalência de SDE, sendo detectadas associações entre as variáveis uso esporádico de álcool e fumo, em relação à ESE. Novos estudos devem ser realizados a fim de prevenir as alterações cognitivas entre os pré-vestibulandos que apresentam SDE ou outro distúrbio do sono.

In vitro and in vivo studies of temozolomide loading in zeolite structures as drug delivery systems for glioblastoma

Zeolites Y (faujasite) and MOR (mordonite) were used as hosts for temozolomide (TMZ), a current good-standard chemotherapeutic agent used in the treatment of glioblastoma brain tumors. TMZ was loaded into zeolites by liquid-phase adsorption at controlled pH. FTIR, 1H NMR, MS, SEM, UV/vis and chemical analysis demonstrated the successful loading of TMZ into zeolite hosts. The hydrolysis of TMZ in MTIC (TMZ metabolite) after the preparation of drug delivery systems (DDS) was observed in simulated body fluid. The effect of zeolites and DDS were evaluated on the viability of glioblastoma cell lines. Unloaded Y zeolite presented toxicity to cancer cells in contrast to MOR. In accordance, the best results in potentiation of the TMZ effect was obtained with MOR. We found that mordonite loaded with 0.026 mmol of TMZ was able to decrease the half maximal inhibitory concentrations (IC50) at least 3-fold in comparison to free temozolomide both in vitro and in vivo.

Tipologia de Lesch em alcoolistas no Brasil

OBJETIVOS: Alcoolismo é uma doença heterogênea, com apresentações clínicas, resultados terapêuticos e recaídas variáveis, indicando vulnerabilidades biológicas diferentes. A Tipologia de Lesch distingue quatro categorias de alcoolismo: Tipo I - graves sintomas de abstinência; Tipo II - álcool como solução para conflitos; Tipo III - álcool para "tratamento" de desordens psiquiátricas; Tipo IV - alterações neurológicas antes do uso de álcool. Este estudo verificou a aplicabilidade de uma classificação do tipo clínico de alcoolismo pela Tipologia de Lesch em um ambulatório público brasileiro de atendimento especializado de alta demanda. MÉTODO: Estudo seccional descritivo, que classificou pacientes do ambulatório de alcoolismo do Hospital Universitário da Universidade Federal do Espírito Santo de acordo com a Tipologia de Lesch. RESULTADOS: A diferenciação pela Tipologia de Lesch foi facilmente conduzida em um serviço ambulatorial público de alta demanda. De 170 pacientes, 21,2% foram classificados como Tipo I; 29,4%, Tipo II; 28,8%, Tipo III e 20,6%, Tipo IV. Embora os diferentes tipos de alcoolismo tenham diferentes apresentações clínicas, o padrão de ingestão alcoólica, idade da primeira ingestão e tempo de abstinência não diferiram entre os tipos de alcoolismo. CONCLUSÃO: A distinção do tipo clínico de alcoolismo de acordo com a Tipologia de Lesch foi considerada aplicável em um ambulatório público brasileiro de grande demanda, sendo os dados encontrados semelhantes aos relatados em estudos realizados em diferentes países. A aplicação dessa classificação poderá definir mudanças nas estratégias de enfrentamento individualizadas do alcoolismo, sendo, entretanto, necessários estudos de seguimento para avaliar os resultados terapêuticos das mesmas.

Synthesis of new peptide derivatives that self-assemble into nanostructured hydrogels for biomedical applications

Tese de Doutoramento em Ciências (área de especialização em Química)

Perfil clínico e sociodemográfico de pacientes com esquizofrenia refratária tratados em um centro terciário

Objetivos A esquizofrenia está associada a alto grau de incapacidade e importantes déficits neuropsicológicos, sociais e vocacionais. Pesquisas têm sido realizadas com o objetivo de identificar fatores preditivos para refratariedade, a fim de melhorar o tratamento e a qualidade de vida do paciente com esquizofrenia. O presente estudo teve o objetivo de verificar a frequência de pacientes com esquizofrenia refratária acompanhados em serviço terciário, estabelecer o perfil clínico e sociodemográfico e analisar possíveis fatores associados à refratariedade clínica. Métodos Sessenta e oito pacientes com esquizofrenia foram incluídos no estudo, sendo 36 refratários ao tratamento (52,9%). Os dados clínicos e sociodemográficos de ambos os grupos foram coletados, analisados e comparados. Um modelo de regressão logística foi elaborado com o objetivo de analisar possíveis fatores associados à refratariedade clínica. Resultados Entre o grupo refratário, houve maior frequência do sexo masculino (p = 0,03), número de antipsicóticos em uso (p < 0,01), internações ao longo da vida (p < 0,01) e de polifarmácia (p < 0,01). Escolaridade, estado civil, história familiar de esquizofrenia e uso de substâncias não foram confirmados como associados à refratariedade. Observou-se atraso temporal entre o estabelecimento da refratariedade clínica e a introdução da clozapina, indicado como o melhor antipsicótico para o tratamento de esquizofrenia refratária. Conclusão É importante e necessário o desenvolvimento de mais pesquisas a fim de investigar possíveis fatores clínicos e sociodemográficos preditores de refratariedade em pacientes com esquizofrenia, objetivando o início mais precoce de ações terapêuticas.

Biosynthesis, detection and toxicology of ochratoxins - optimisation of production

Ochratoxin A (OTA) is a very well known mycotoxin found in several food commodities for which maximum limits are being discussed in EC in other to produce appropriate regulations. OTA is one of several ochratoxins produced by Aspergillus and Penicillium species. All the compounds in this group have a molecular structure very similar to OTA and some were already isolated from natural substrates. Several of these compounds such as ochratoxin , methyl and ethyl ester of ochratoxin A, 4-R and S-hydroxyochratoxin A, 10-hydroxyochratoxin A and ochratoxin A open lactone are commercially unavailable. However, they can be easily synthesized through OTA modification. With the main objective of its application on further research works, OTA production, isolation and purification has been optimised from an A. alliaceus strain grown on wheat medium. Synthesis and purification of some OTA derivatives has been achieved and an HPLC method for their detection was optimised. Data about their production by several species of Aspergillus will be presented. The toxicological properties of ochratoxins are still not very clear and a future EC safety limit for OTA will depend on e.g., a better clarification of its carcinogenity. Could OTA derivatives play a role here?

New bio-strategies for ochratoxin A detoxification using lactic acid bacteria

The occurrence of mycotoxigenic moulds such as Aspergillus, Penicillium and Fusarium in food and feed has an important impact on public health, by the appearance of acute and chronic mycotoxicoses in humans and animals, which is more severe in the developing countries due to lack of food security, poverty and malnutrition. This mould contamination also constitutes a major economic problem due the lost of crop production. A great variety of filamentous fungi is able to produce highly toxic secondary metabolites known as mycotoxins. Most of the mycotoxins are carcinogenic, mutagenic, neurotoxic and immunosuppressive, being ochratoxin A (OTA) one of the most important. OTA is toxic to animals and humans, mainly due to its nephrotoxic properties. Several approaches have been developed for decontamination of mycotoxins in foods, such as, prevention of contamination, biodegradation of mycotoxins-containing food and feed with microorganisms or enzymes and inhibition or absorption of mycotoxin content of consumed food into the digestive tract. Some group of Gram-positive bacteria named lactic acid bacteria (LAB) are able to release some molecules that can influence the mould growth, improving the shelf life of many fermented products and reducing health risks due to exposure to mycotoxins. Some LAB are capable of mycotoxin detoxification. Recently our group was the first to describe the ability of LAB strains to biodegrade OTA, more specifically, Pediococcus parvulus strains isolated from Douro wines. The pathway of this biodegradation was identified previously in other microorganisms. OTA can be degraded through the hydrolysis of the amide bond that links the L-β-phenylalanine molecule to the ochratoxin alpha (OTα) a non toxic compound. It is known that some peptidases from different origins can mediate the hydrolysis reaction like, carboxypeptidase A an enzyme from the bovine pancreas, a commercial lipase and several commercial proteases. So, we wanted to have a better understanding of this OTA degradation process when LAB are involved and identify which molecules where present in this process. For achieving our aim we used some bioinformatics tools (BLAST, CLUSTALX2, CLC Sequence Viewer 7, Finch TV). We also designed specific primers and realized gene specific PCR. The template DNA used came from LAB strains samples of our previous work, and other DNA LAB strains isolated from elderberry fruit, silage, milk and sausages. Through the employment of bioinformatics tools it was possible to identify several proteins belonging to the carboxypeptidase family that participate in the process of OTA degradation, such as serine type D-Ala-D-Ala carboxypeptidase and membrane carboxypeptidase. In conclusions, this work has identified carboxypeptidase proteins being one of the molecules present in the OTA degradation process when LAB are involved.

Influence of chitosan coating on protein-based nanohydrogels properties and in vitro gastric digestibility

Chitosan coating was applied in Lactoferrin (Lf)-Glycomacropeptide (GMP) nanohydrogels by layer-by-layer coating process. A volume ratio of 0.1 of Lf-GMP nanohydrogels (0.2 mg.mL-1, at pH 5.0) to chitosan (1 mg.mL-1, at pH 3) demonstrated to be the optimal condition to obtain stable nanohydrogels with size of 230 ± 12 nm, a PdI of 0.22 ± 0.02 and a -potential of 30.0 ± 0.15 mV. Transmission electron microscopy (TEM) images showed that the application of chitosan coating in Lf-GMP did not affect the spherical shape of nanohydrogels and confirmed the low aggregation of nanohydrogels in solution. The analysis of chemical interactions between chitosan and Lf-GMP nanohydrogels were performed by Fourier transform infrared spectroscopy (FTIR) and by circular dichroism (CD) that revealed that a specific chemical interaction occurring between functional groups of protein-based nanohydrogels and active groups of the chitosan was established. The effect of chitosan coating on release mechanisms of Lf-GMP nanohydrogels at acid conditions (pH 2, 37 ºC) was evaluated by the encapsulation of a model compound (caffeine) in these systems. Linear Superposition Model was used to fit the experimental data and revealed that Fick and relaxation mechanisms are involved in caffeine release. It was also observed that the Fick contribution increase with the application of chitosan coating. In vitro gastric digestion was performed with Lf-GMP nanohydrogels and Lf-GMP nanohydrogels with chitosan coating and it was observed that the presence of chitosan improve the stability of Lf and GMP (proteins were hydrolysed at a slower rate and were present in solution by longer time). Native electrophoreses revealed that the nanohydrogels without coating remained intact in solution until 15 min and with chitosan coating remained intact until 60 min, during gastric digestion.

Conversion of Cn-unsaturated into Cn-2-saturated LCFA can occur uncoupled from methanogenesis in anaerobic bioreactors

Fat, oils, and grease present in complex wastewater can be readily converted to methane, but the energy potential of these compounds is not always recyclable, due to incomplete degradation of long chain fatty acids (LCFA) released during lipids hydrolysis. Oleate (C18:1) is generally the dominant LCFA in lipid-containing wastewater, and its conversion in anaerobic bioreactors results in palmitate (C16:0) accumulation. The reason why oleate is continuously converted to palmitate without further degradation via β-oxidation is still unknown. In this work, the influence of methanogenic activity in the initial conversion steps of unsaturated LCFA was studied in 10 bioreactors continuously operated with saturated or unsaturated C16- and C18-LCFA, in the presence or absence of the methanogenic inhibitor bromoethanesulfonate (BrES). Saturated Cn-2-LCFA accumulated both in the presence and absence of BrES during the degradation of unsaturated Cn-LCFA, and represented more than 50\% of total LCFA. In the presence of BrES further conversion of saturated intermediates did not proceed, not even when prolonged batch incubation was applied. As the initial steps of unsaturated LCFA degradation proceed uncoupled from methanogenesis, accumulation of saturated LCFA can be expected. Analysis of the active microbial communities suggests a role for facultative anaerobic bacteria in the initial steps of unsaturated LCFA biodegradation. Understanding this role is now imperative to optimize methane production from LCFA.