Comparison of vaginal microbiota fingerprints from healthy and bacterial vaginosis-positive portuguese women

Bacterial Vaginosis (BV) is a common disease in women of reproductive age and is characterized by the substitution of Lactobacillus species,which are predominant in the normal vaginal microbiota,by rapidly proliferating anaerobic bacteria, particularly Gardnerellavaginalis. The aim of this study was to study microbial communities’ structure in the vaginal microbiota of healthy and BV-positive Portuguese women. To this end, DNA obtained from vaginal samples of 22 BV-negative and 19 BV-positive women was analyzed using a PCR-DGGE approach.Total bacterial communities were amplified using general 16S rRNA gene primers. Group-specific primers were also used targeting Lactobacillus and Bifidobacterium genera and G.vaginalis. DGGE profiles were compared using the BioNumericsTM software package (Applied Maths, Belgium). Similarity between DGGE profiles was determined by calculating similarity indices of the densitometric curves of the compared profiles, using the Dice product-moment correlation. Different DGGE profiles could be obtained for BV-positive and BV-negative samples and this was verified for all primers sets utilized, suggesting that alteration of microbial community structure of BV-positive and -negative samples could be detected by PCR-DGGE. DGGE profiles obtained from samples of BV-positive women were more diverse that the ones from healthy women (as determined by a higher number of DGGE bands). The analysis of the standard electrophoretic bands for bacteria reveals an intrinsic diversity even within the two groups studied: similarities in bacterial DGGE profiles vary between 14- 78% and 47-100% in BV-positive and BV-negative samples, respectively. Among the 19 BV-positive women studied 18 were colonized with G. vaginalis. G.vaginalis was not detected in any of the healthy women samples. The analysis of Lactobacillus communities revealed a higher diversity in BV-negative women than in BV-positive ones, which confirms the association of Lactobacillus in healthy vaginal microbial communities. A more thoroughly comparison between BV-negative and BV-positive, including the retrieval of sequencing data from these samples, is necessary for getting more insight on BV influence on vaginal microbiota

Development of bacterial cellulose wound dressings with controlled delivery of vitamin D3

Book of Abstracts of CEB Annual Meeting 2017[Excerpt] Wounds, in particular traumatic (e.g. burns) and chronic ones, are a major cause of morbidity and impaired life quality. They often result in long hospitalization stays, taking up substantial health resources in developed countries. This proposal aims at developing a safe, easy-to-use and nonexpensive approach to efficiently address this problem, by attaining faster and proper wound healing. Recent studies showed that an antimicrobial peptide (AMP), LLKKK18, released from conjugates with dextrin embedded in a Carbopol hydrogel significantly improved burn wound healing. In addition to antimicrobial activity, this peptide stimulates vascularization, thus supporting a faster healing and tissue regeneration[1]. As such, one can hypothesize that a hydrogel comprising drugs that stimulate the expression of LL37 will improve wound healing while keeping the wound area infection-free. This work comprised the approach towards the development of a novel bacterial nanocellulose (BNC) dressing. BNC, already used clinically for the treatment of burn wounds due to the unique properties like high water holding capacity, high crystallinity, ultrafine fiber network, high resistance, high moldability and biocompatibility[2]. In this work BNC will be used as drug carriers for the controlled release of drugs, namely of vitamin D3, an inducer of an endogenous expression of AMP LL37, known for accelerating the wound healing process, and as a protective barrier against exogenous agents (dust, microorganism) that can impair wound healing. [...]info:eu-repo/semantics/publishedVersio

Squaraine dyes derived from indolenine and benzo[e]indole as potential fluorescent probes for HSA detection and antifungal agents

Four squaraine dyes derived from 2,3,3-trimethylindolenine and 1,1,2-trimethyl-1H-benzo[e]indole with different combinations of barbituric groups attach to the central ring, having ester groups and alkyl chains in the nitrogen atoms of heterocyclic rings were synthesized. These dyes were fully characterized and their photophysical behavior was studied in ethanol and phosphate-buffered saline solution. Absorption and emission bands between 631 and 712 nm were detected, with the formation of aggregates in aqueous media, which is typical of this class of dyes. Tests carried out with 1,3-diphenylisobenzofuran allowed us to verify the ability of the dyes to produce singlet oxygen. The interaction of synthesized dyes with human serum albumin (HSA) was also evaluated, being demonstrated a linear correlation between fluorescence intensity and protein concentration. The antifungal potential of the dyes against the yeast Saccharomyces cerevisiae was evaluated using a broth microdilution assay. In order to test the photosensitizing capacity of the synthesized dyes, tests were carried out in the dark and with irradiation, using a custom-built light-emitting diode that emits close to the absorption wavelength of the studied dyes. The results showed that the interaction of dyes with HSA and the antifungal activity depends on the different structural modifications of the dyes.We thanks to Fundação para a Ciência e Tecnologia (FCT), Comissão de Coordenação e Desenvolvimento Regional do Norte (CCDR-N) and FEDER (European Fund for Regional Development)-COMPETEQREN-EU for financial support to the research centers CQ/UM (UIDB/00686/2020), CBMA (UID/BIA/04050/2020), CQ/VR (UID/QUI/UI0616/2019) and CICSUBI (POCI-01-0145-FEDER-007491), as well as PhD grants to V.S.D.G. (UMINHO/BD/43/2016) and J.C.C.F. (SFRH/BD/133207/2017)

Structural differences on cell wall polysaccharides of brewer's spent Saccharomyces and microarray binding profiles with immune receptors

Funding Information: This work was financial supported by FCT - Fundação para a Ciência e a Tecnologia , I.P. within the project “Yeast4FoodMed” ( POCI-01-0145-FEDER-030936 and PTDC/BAA-AGR/30936/2017 ) and LAQV/REQUIMTE ( UIDB/50006/2020 and UIDP/50006/2020 ) through national funds and, where applicable, co-financed by the FEDER , within the PT2020 Partnership Agreement. This work was also financed by national funds from FCT , in the scope of the project UIDP/04378/2020 and UIDB/04378/2020 of the Research Unit on Applied Molecular Biosciences - UCIBIO and the project LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy - i4HB. Rita Bastos ( PD/BD/114579/2016 ), Viviana G. Correia ( PD/BD/105727/2014 ) and Angelina S. Palma ( PTDC/BIA-MIB/31730/2017 ) were supported by FCT . Elisabete Coelho (CDL-CTTRI-88-ARH/2018 - REF. 049-88-ARH/2018 ) and Benedita Pinheiro thank the research contracts funded by FCT through program DL 57/2016 – Norma transitória. Publisher Copyright: © 2022 The AuthorsBrewing practice uses the same yeast to inoculate the following fermentation (repitching). Saccharomyces pastorianus, used to produce Lager beer, is widely reused, not changing its fermentation performance. However, S. cerevisiae, used to produce Ale beer, is partial or not even reused, due to its poor performance. It is hypothesized that cells modulate their wall polysaccharides to increase the cell-wall strength. In this work industrial S. cerevisiae and S. pastorianus brewer's spent yeasts with different repitching numbers were studied. Glucans were the main component of S. cerevisiae whereas mannoproteins were abundant in S. pastorianus. The major changes were noticed on glucans of both species, β1,3-glucans decrease more pronounced in S. cerevisiae. The increase of α1,4-Glc, related with osmotolerance, was higher in S. cerevisiae while β1,4-Glc, related with cell-wall strength, had a small increase. In addition, these structural details showed different binding profiles to immune receptors, important to develop tailored bioactive applications.publishersversionpublishe

Injectable hyaluronic acid bionanocomposite hydrogels: from biomaterial development to biological performance outcomes

Introduction: Injectable hyaluronic acid (HA) hydrogels have been increasingly applied in tissue engineering (TE) envisioning minimal invasive approaches. However, traditional HA hydrogels lack structural integrity that makes them less competitive in strategies where good mechanical properties are required. Here we propose the use of cellulose nanocrystals (CNCs), the nature â carbon nanotubesâ , as nanofillers and crosslinkers in a fully biobased strategy for the production of reinforced HA nanocomposite hydrogels[1]. Due to their distinct mechanical properties, biocompatibility and excellent aqueous colloidal stability, CNCs are being increasingly considered in hydrogel development targeting biomedical applications[2]. We hypothesise that besides structural reinforcement, in TE strategies the CNCâ s surface SO3-groups may also potentially act as semisynthetic mimicry of ECM sulfated glycosaminoglycans, which are known to induce and control specific cell functions on the cellular microenvironment through interactions with soluble biomolecules, e.g. proteins, growth factors (GFs)[3]. Materials and Methods: In situ crosslinkable and injectable hydrogels were prepared based on hydrazone coupling of adipic acid dihydrazide-modified HA (ADH-HA) and aldehyde-modified HA (a-HA), reinforced with aldehyde-modified CNCs (a-CNCs) (Figure 1). The hydrogel precursors were fully characterized by several spectroscopic, chromatographic, and imaging techniques, and the hydrogels were characterized in terms of internal morphology, mechanical properties, swelling and degradation behaviour in the presence of hyaluronidase. The biological performance of the developed nanocomposites was assessed towards human adipose derived stem cells (hASCs). Results and Discussion: The incorporation of a-CNCs in the hydrogelâ s network had a remarkable impact over the physical and biological performance of the injectable biomaterial. Nanocomposite hydrogels showed improved microstructure and mechanical properties (increased Eâ  up to 2.7-fold compared to unfilled hydrogels), lower equilibrium swelling ratios and higher resistance to bulk hyaluronidase degradation. HA-CNCs exhibited preferential cell supportive properties in in vitro culture conditions, in both surface cell seeding and cell encapsulation tests. Particularly, hASCs encapsulated in HA-CNCs hydrogels demonstrated ability to spread within the volume of gels and exhibited pronounced proliferative activity. This impact over cellâ s behaviour is correlated with the higher structural integrity of the hydrogel matrix and potential interaction of soluble microenvironmental cues with the CNCâ s surface sulphate groups. Conclusions: The proposed strategy demonstrated to be a valuable approach for fine tuning the structural, biomechanical and biochemical properties of injectable HA hydrogels.  The combined effects of enhanced stability and mechanical properties with the incorporation of mimetic ECM biochemical cues in HA-CNCs hydrogels, proved to positively impact their biological performance for TE applications. Considering the promising outcomes, we are currently exploring the potential of the developed system when combined with discrete GFs or the GFs pool from platelet lysates in specific TE strategies

How can biomolecules improve mucoadhesion of oral insulin? A comprehensive insight using ex-vivo, in silico and in vivo models

Currently, insulin can only be administered through the subcutaneous route. Due to the flaws associated with this route, it is of interest to orally deliver this drug. However, insulin delivered orally has several barriers to overcome as it is degraded by the stomach’s low pH, enzymatic content, and poor absorption in the gastrointestinal tract. Polymers with marine source like chitosan are commonly used in nanotechnology and drug delivery due to their biocompatibility and special features. This work focuses on the preparation and characterization of mucoadhesive insulin-loaded polymeric nanoparticles. Results showed a suitable mean size for oral administration (<600 nm by dynamic laser scattering), spherical shape, encapsulation efficiency (59.8%), and high recovery yield (80.6%). Circular dichroism spectroscopy demonstrated that protein retained its secondary structure after encapsulation. Moreover, the mucoadhesive potential of the nanoparticles was assessed in silico and the results, corroborated with ex-vivo experiments, showed that using chitosan strongly increases mucoadhesion. Besides, in vitro and in vivo safety assessment of the final formulation were performed, showing no toxicity. Lastly, the insulin-loaded nanoparticles were effective in reducing diabetic rats’ glycemia. Overall, the coating of insulin-loaded nanoparticles with chitosan represents a potentially safe and promising approach to protect insulin and enhance peroral delivery.Supported in part by UID/DTP/04138/2019 from FCT, Portugal and DREAMS (ULHT). SEM analysis was funded by FCT/MCTES for the financial support to CESAM (UIDP/50017/2020+UIDB/50017/2020), through national funds. Acknowledgments: The authors are grateful to the Carla Vânia (iMedUlisboa) for her collaboration in HPLC analysis and Joana Moreira (ECTS-ULHT) for her collaboration in conducting some experiments.info:eu-repo/semantics/publishedVersio

How can biomolecules improve mucoadhesion of oral insulin? A comprehensive insight using ex-vivo, in silico, and in vivo models

Funding: Supported in part by UID/DTP/04138/2019 from FCT, Portugal and DREAMS (ULHT). SEM analysis was funded by FCT/MCTES for the financial support to CESAM (UIDP/50017/2020+UIDB/50017/2020), through national funds.Currently, insulin can only be administered through the subcutaneous route. Due to the flaws associated with this route, it is of interest to orally deliver this drug. However, insulin delivered orally has several barriers to overcome as it is degraded by the stomach’s low pH, enzymatic content, and poor absorption in the gastrointestinal tract. Polymers with marine source like chitosan are commonly used in nanotechnology and drug delivery due to their biocompatibility and special features. This work focuses on the preparation and characterization of mucoadhesive insulin-loaded polymeric nanoparticles. Results showed a suitable mean size for oral administration (<600 nm by dynamic laser scattering), spherical shape, encapsulation efficiency (59.8%), and high recovery yield (80.6%). Circular dichroism spectroscopy demonstrated that protein retained its secondary structure after encapsulation. Moreover, the mucoadhesive potential of the nanoparticles was assessed in silico and the results, corroborated with ex-vivo experiments, showed that using chitosan strongly increases mucoadhesion. Besides, in vitro and in vivo safety assessment of the final formulation were performed, showing no toxicity. Lastly, the insulin-loaded nanoparticles were effective in reducing diabetic rats’ glycemia. Overall, the coating of insulin-loaded nanoparticles with chitosan represents a potentially safe and promising approach to protect insulin and enhance peroral delivery.publishersversionpublishe

Improved Efficiency of Brewer’s Spent Grain Arabinoxylans by Ultrasound-Assisted Extraction

Arabinoxylan (AX) rich extracts from brewer’s spent grain (BSG) were produced by the application of ultrasound-assisted extraction (UAE) and conventional alkaline extraction (AKE). UAE and AKE were optimised for the production of the highest yield of ethanol insoluble material using response surface methodology (RSM). The efficiency of UAE was established by the significant reduction of time (7h to 25 min) and energy when compared to AKE, to recover similar amount of AX (60%) from BSG, leading to the production of starch-free AX-rich extracts

Drosophila Genes That Affect Meiosis Duration Are among the Meiosis Related Genes That Are More Often Found Duplicated

Using a phylogenetic approach, the examination of 33 meiosis/meiosis-related genes in 12 Drosophila species, revealed nine independent gene duplications, involving the genes cav, mre11, meiS332, polo and mtrm. Evidence is provided that at least eight out of the nine gene duplicates are functional. Therefore, the rate at which Drosophila meiosis/meiosis-related genes are duplicated and retained is estimated to be 0.0012 per gene per million years, a value that is similar to the average for all Drosophila genes. It should be noted that by using a phylogenetic approach the confounding effect of concerted evolution, that is known to lead to overestimation of the duplication and retention rate, is avoided. This is an important issue, since even in our moderate size sample, evidence for long-term concerted evolution (lasting for more than 30 million years) was found for the meiS332 gene pair in species of the Drosophila subgenus. Most striking, in contrast to theoretical expectations, is the finding that genes that encode proteins that must follow a close stoichiometric balance, such as polo, mtrm and meiS332 have been found duplicated. The duplicated genes may be examples of gene neofunctionalization. It is speculated that meiosis duration may be a trait that is under selection in Drosophila and that it has different optimal values in different species