PHB-HV 3D Scaffold supports osteoblast growth

Bone tissue engineering requires a biocompatible scaffold that supports cell growth and enhances the native repair process. Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHB-HV) is a biodegradable 3D scaffold with 88.1 â 0.3% porosity and pore size of 163.5 â 0.1 mm. Previous studies demonstrated the potential of PHB-HV as a scaffold in spinal cord repair. The aim of this study was to evaluate PHB-HV as a scaffold for bone regeneration by assessing the cytocompatability of this scaffold.

Quantitative image analysis as a tool for Yarrowia lipolytica dimorphic growth evaluation in different culture media

Yarrowia lipolytica, a yeast strain with a huge biotechnological potential, capable to produce metabolites such as γ-decalactone, citric acid, intracellular lipids and enzymes, possesses the ability to change its morphology in response to environmental conditions. In the present study, a quantitative image analysis (QIA) procedure was developed for the identification and quantification of Y. lipolytica W29 and MTLY40-2P strains dimorphic growth, cultivated in batch cultures on hydrophilic (glucose and N-acetylglucosamine (GlcNAc) and hydrophobic (olive oil and castor oil) media. The morphological characterization of yeast cells by QIA techniques revealed that hydrophobic carbon sources, namely castor oil, should be preferred for both strains growth in the yeast single cell morphotype. On the other hand, hydrophilic sugars, namely glucose and GlcNAc caused a dimorphic transition growth towards the hyphae morphotype. Experiments for γ-decalactone production with MTLY40-2P strain in two distinct morphotypes (yeast single cells and hyphae cells) were also performed. The obtained results showed the adequacy of the proposed morphology monitoring tool in relation to each morphotype on the aroma production ability. The present work allowed establishing that QIA techniques can be a valuable tool for the identification of the best culture conditions for industrial processes implementation.

Study of the potential employment of Malvaceae Species in composites materials

The employ of vegetal fibers for textiles and composites represents a great potential in economic and social sustainable development. Some Malvaceae species are considered tropical cosmopolitans, such as from Sida genus. Several species of this genus provide excellent textile bast fibers, which are very similar in qualities to the jute textile fiber. The objective of the present study is present the physicochemical characterization of six Brazilian vegetal fibers: Sida rhombifolia L.; Sida carpinifolia L. f.; Sidastrum paniculatum (L.) Fryxell; Sida cordifolia L.; Malvastrum coromandelianum (L.) Gurck; Wissadula subpeltata (Kuntze) R.E.Fries. Respectively the two first species are from Brazilian Atlantic Forest biome and the four remaining from Brazilian Cerrado biome, despite of present in other regions of the planet. The stems of these species were retted in water at 37oC for 20 days. The fibers were tested in order to determine tensile rupture strength, tenacity, elongation, Young’s modulus, cross microscopic structure, Scanning Electronic Microscopy (SEM), regain, combustion, acid, alkali, organic solvent and cellulase effects, pH of the aqueous extract, Differential Scanning Calorimetry (DSC) and Thermogravimetric Analysis (TGA). The obtained values were compared with those from fibers of recognized applicability in the textile industry including hemp. The results are promising in terms of their employment in thermoset and thermoplastic medium resistance composites.

IL-17A promotes intracellular growth of Mycobacterium by inhibiting apoptosis of infected macrophages

The fate of infected macrophages is a critical aspect of immunity to mycobacteria. By depriving the pathogen of its intracellular niche, apoptotic death of the infected macrophage has been shown to be an important mechanism to control bacterial growth. Here, we show that IL-17 inhibits apoptosis of Mycobacterium bovis BCG- or Mycobacterium tuberculosis-infected macrophages thus hampering their ability to control bacterial growth. Mechanistically, we show that IL-17 inhibits p53, and impacts on the intrinsic apoptotic pathway, by increasing the Bcl2 and decreasing Bax expression, decreasing cytochrome c release from the mitochondria, and inhibiting caspase-3 activation. The same effect of IL-17 was observed in infected macrophages upon blockade of p53 nuclear translocation. These results reveal a previously unappreciated role for the IL-17/p53 axis in the regulation of mycobacteria-induced apoptosis and can have important implications in a broad spectrum of diseases where apoptosis of the infected cell is an important host defense mechanism.

Biomimetic supramolecular designs for the controlled release of growth factors in bone regeneration

The extracellular matrix (ECM) of tissues is an assembly of insoluble macromolecules that specifically interact with soluble bioactive molecules and regulate their distribution and availability to cells. Recapitulating this ability has been an important target in controlled growth factor delivery strategies for tissue regeneration and requires the design of multifunctional carriers. This review describes the integration of supramolecular interactions on the design of delivery strategies that encompass self-assembling and engineered affinity components to construct advanced biomimetic carriers for growth factor delivery. Several glycan- and peptide-based self-assemblies reported in the literature are highlighted and commented upon. These examples demonstrate how molecular design and chemistry are successfully employed to create versatile multifunctional molecules which self-assemble/disassemble in a precisely predicted manner, thus controlling compartmentalization, transport and delivery. Finally, we discuss whether recent advances in the design and preparation of supramolecular delivery systems have been sufficient to drive real translation towards a clinical impact. 

Crowdfunding e resiliência das comunidades: estudo de caso

Dissertação de mestrado integrado em Engenharia e Gestão de Sistemas de Informação

Targeting lactate transport suppresses in vivo breast tumour growth.

BACKGROUND Most cancers, including breast cancer, have high rates of glucose consumption, associated with lactate production, a process referred as "Warburg effect". Acidification of the tumour microenvironment by lactate extrusion, performed by lactate transporters (MCTs), is associated with higher cell proliferation, migration, invasion, angiogenesis and increased cell survival. Previously, we have described MCT1 up-regulation in breast carcinoma samples and demonstrated the importance of in vitro MCT inhibition. In this study, we performed siRNA knockdown of MCT1 and MCT4 in basal-like breast cancer cells in both normoxia and hypoxia conditions to validate the potential of lactate transport inhibition in breast cancer treatment. RESULTS The effect of MCT knockdown was evaluated on lactate efflux, proliferation, cell biomass, migration and invasion and induction of tumour xenografts in nude mice. MCT knockdown led to a decrease in in vitro tumour cell aggressiveness, with decreased lactate transport, cell proliferation, migration and invasion and, importantly, to an inhibition of in vivo tumour formation and growth. CONCLUSIONS This work supports MCTs as promising targets in cancer therapy, demonstrates the contribution of MCTs to cancer cell aggressiveness and, more importantly, shows, for the first time, the disruption of in vivo breast tumour growth by targeting lactate transport.

Looking for mechanisms regulating lung growth in CDH: rat and human studies

Tese de Doutoramento em Ciências da Saúde

Growth of sulfate reducing bacteria on the methanogenic inhibitor BES

The metabolism of methanogenic archaea is inhibited by 2-bromoethanesulfonate (BES). Methane production is blocked because BES is an analog of methyl-coenzyme M and competes with this key molecule in the last step of methanogenesis. For this reason, BES is commonly used in several studies to avoid growth of acetoclastic and hydrogenotrophic methanogens [1]. Despite its effectiveness as methanogenic inhibitor, BES was found to alter microbial communities’ structure, to inhibit the metabolism of non-methanogenic microorganisms and to stimulate homoacetogenic metabolism [2,3]. Even though sulfonates have been reported as electron acceptors for sulfate- and sulfite-reducing bacteria (SRB), only one study described the reduction of BES by complex microbial communities [4]. In this work, a sulfate-reducing bacterium belonging to Desulfovibrio genus (98 % identity at the 16S rRNA gene level with Desulfovibrio aminophilus) was isolated from anaerobic sludge after several successive transfers in anaerobic medium containing BES as sole substrate. Sulfate was not supplemented to the anaerobic growth medium. This microorganism was able to grow under the following conditions: on BES plus H2/CO2 in bicarbonate buffered medium; on BES without H2/CO2 in bicarbonate buffered medium; and on BES in phosphate buffered medium. The main products of BES utilization were sulfide and acetate, the former was produced by the reduction of sulfur from the sulfonate moiety of BES and the latter likely originated from the carbon backbone of the BES molecule. BES was found, in this study, to represent not only an alternative electron acceptor but also to serve as electron donor, and sole carbon and energy source, supporting growth of a Desulfovibrio sp. obtained in pure culture. This is the first study that reports growth of SRB with BES as electron donor and electron acceptor, showing that the methanogenic inhibitor is a substrate for anaerobic growth.

The effect of hydrodynamic conditions in Corynebacterium glutamicum growth

[Excerpt] Corynebacterium glutamicum is a facultative anaerobic, gram-positive bacterium with a GRAS status that grows fast and achieves high cell densities. C. glutamicum is commonly used in amino acids production, and is also able to convert sugars in organic acids (OA) and alcohols in specific conditions: anaerobic and limited-oxygen environments. In these conditions, the carbon metabolism is modified, namely the flux shifts from the pentose phosphate pathway to glycolysis and the TCA cycle flux decreases and consequently bacterial growth is strongly affected [1,2]. (...)

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.

Novas bio-estratégias para a destoxificação de ocratoxina A utilizando bactérias do ácido láctico

A ocorrência de bolores micotoxigénicos pertencentes aos géneros Aspergillus, Penicillium e Fusarium em alimentos para consumo Humano e animal, tem um impacto importante sobre a saúde pública e constitui também um importante problema económico. Isto é devido à síntese por este tipo de fungos filamentosos de metabolitos altamente tóxicos conhecidos como micotoxinas. A maioria das micotoxinas são substâncias cancerígenas, mutagénicas, neurotóxicas e imunossupressoras, sendo a ocratoxina A (OTA) uma das mais importantes. A OTA é uma micotoxina, tóxica para os animais e Humanos principalmente devido às suas propriedades nefrotóxicas. Alguns grupos de bactérias gram positivas nomeadamente as bactérias do ácido láctico (BAL) são capazes de controlar o crescimento de fungos, melhorando e aumentando a vida útil de muitos produtos fermentados e, assim, reduzir os riscos para a saúde provocados pela exposição às micotoxinas. Algumas BAL são, também, capazes de destoxificar certas micotoxinas. Em trabalhos anteriores do nosso grupo foi observada a biodegradação da OTA por estirpes de Pediococcus parvulus isoladas de vinhos do Douro. Assim, com este trabalho, pretendeu-se compreender com maior detalhe o processo de biodegradação da OTA pelas referidas estirpes e identificar quais as enzimas que estão associadas à sua biodegradação. Para atingir este objetivo utilizaram-se algumas ferramentas ioinformáticas (BLAST, CLUSTALX2, CLC Sequence Viewer 7, Finch TV), desenharam-se primers específicos e realizaram-se PCR específicos para os genes envolvidos. Através da utilização de ferramentas de bioinformática, foi possível identificar várias proteínas que pertencem à família das carboxipeptidases e que podem eventualmente participar no processo da degradação da OTA, tais como D-Ala-D-Ala carboxipeptidase serínica e carboxipeptidase membranar. Estas BAL podem desempenhar um papel importante na destoxificação da OTA, sendo as carboxipeptidases uma das enzimas envolvidas na sua biodegradação.

Study of the retinoic acid regulated genes in the rodent hypothalamus controlling growth and appetite and the potential for the epigenetic control

Dissertação de mestrado em Bioquímica (área de especialização em Biomedicina)

Psychometric properties of the Portuguese version of the Posttraumatic Growth Inventory Short Form among divorced adults

The aim of this study was to develop and validate a Portuguese version of the Short Form of the Posttraumatic Growth Inventory (PTGI-SF). Using an online convenience sample of Portuguese divorced adults (N = 482), we confirmed the oblique five-factor structure of the PTGI-SF by confirmatory factor analysis. The results demonstrated the measurement invariance across divorce initiator status groups. Total score and factors of PTGI-SF showed good internal consistency, with the exception of the New Possibilities factor, which revealed an acceptable reliability. The Portuguese PTGI-SF showed a satisfactory convergent validity. In terms of discriminant validity, posttraumatic growth assessed by the Portuguese PTGI-SF was a distinct factor from posttraumatic psychological adjustment. These preliminary findings suggest the cultural adaptation and also psychometric properties of the present Portuguese PTGI-SF to measure posttraumatic growth after personal crisis.

Mother’s anxiety and depression and associated risk factors during early pregnancy: effects on fetal growth and activity at 20–22 weeks of gestation

To examine effects of mother's anxiety and depression and associated risk factors during early pregnancy on fetal growth and activity. Repeated measures of mother's anxiety (State-Anxiety Inventory (STAI-S)) and depression (Edinburgh Postnatal Depression Scale (EPDS)) and related socio demographics and substance consumption were obtained at the 1st and 2nd pregnancy trimesters, and fetus' (N = 147) biometric data and behavior was recorded during ultrasound examination at 20-22 weeks of gestation. Higher anxiety symptoms were associated to both lower fetal growth and higher fetal activity. While lower education, primiparity, adolescent motherhood, and tobacco consumption predicted lower fetal growth, coffee intake predicted lower fetal activity. Vulnerability of fetal development to mother's psychological symptoms as well as to other sociodemographic and substance consumption risk factors during early and mid pregnancy is suggested.