Deri yara kaplamalarindaki uygulamalar için elektrik alan lif çekim ile üretilmiş Tiger 17 peptit yüklü polikaprolakton/selüloz asetat nano lifli matlar

A skin wound if not properly treated can result in a chronic wound susceptible to widespread infections, which can result in the patient's death. Currently, tissue engineering is described as an interdisciplinary field that combines principles of engineering, chemistry and biology to generate solutions that allow to repair, restore and/or improve the functions of injured tissues. In the same sense, the textile area addresses solutions based on polymeric fibers, produced from a wide range of polymers, which allow the generation of structures with a large surface area, porosity and mechanical resistance that can be used as bioactive dressings that promote a healing and efficient antimicrobial activity. This research work focused on the synthesis of Tiger 17, through microwave-assisted solid-phase synthesis methodologies, and Tiger 17 commercially obtained, respective structural characterization and evaluation of the antimicrobial capacity. Simultaneously, nanofibrous polymer matrices were produced using the electrospinning technique with the aim of immobilizing the developed biomolecule and thus creating potential vehicles for a local and sustainable antimicrobial action (controlled release). In order to verify its physical and chemical properties, advanced characterization techniques were used: proton nuclear magnetic resonance, high performance liquid chromatography, optical microscopy, scanning electron microscopy, fourier transform infrared spectroscopy–attenuated total reflectance, thermogravimetry, differential scanning calorimetry, contact angle and surface energy and determination of porosity and hydration.Düzgün tedavi edilmeyen bir cilt yarası, hastanın ölümüyle sonuçlanabilecek yaygın enfeksiyonlara duyarlı kronik bir yaraya neden olabilir. Günümüzde doku mühendisliği, hasarlı dokuların işlevlerini onarmaya, eski haline getirmeye ve/veya iyileştirmeye olanak tanıyan çözümler üretmek için mühendislik, kimya ve biyolojinin ilkelerini birleştiren disiplinler arası bir alan olarak tanımlanmaktadır. Bu kapsamda, tekstil, iyileşmeyi ve etkili antimikrobiyal aktiviteyi teşvik eden biyoaktif pansuman olarak kullanılabilecek, geniş yüzey alanlı, gözenekli ve mekanik dirençli yapıların oluşturulmasına izin veren, geniş bir polimer yelpazesinden üretilen liflere dayalı çözümler sunmaktadır. Bu araştırma çalışması, mikrodalga destekli katı faz sentez metodolojileri aracılığıyla Tiger 17'nin sentezine ve ticari olarak temin edilen Tiger 17 ile birlikte yapısal karakterizasyonunun ve antimikrobiyal kapasitenin değerlendirilmesine odaklanmıştır. Aynı zamanda, geliştirilen biyomolekülü hareketsiz hale getirmek ve böylece lokal ve sürdürülebilir bir antimikrobiyal etki (kontrollü salım) için potansiyel çözümler yaratmak amacıyla elektrik alan lif üretim tekniği kullanılarak nanolifli polimer matrisler üretildi. Yapının fiziksel ve kimyasal özelliklerini doğrulamak için, proton nükleer manyetik rezonans, yüksek performanslı sıvı kromatografisi, optik mikroskopi, taramalı elektron mikroskobu, fourier dönüşümü kızılötesi spektroskopisi-zayıflatılmış toplam yansıma, termogravimetri, diferansiyel taramalı kalorimetri, temas açısı, yüzey enerjisi, gözeneklilik ve su tutma ölçümleri gibi ileri karakterizasyon teknikleri kullanıldı

How additive manufacturing can boost the bioactivity of baked functional foods

The antioxidant activity of baked foods is of utmost interest when envisioning enhancing their health benefits. Incorporating functional ingredients is challenging since their bioactivity naturally declines during baking. In this study, 3D food printing and design of experiments are employed to clarify how the antioxidant activity of cookies enriched with encapsulated polyphenols can be maximized. A synergistic effect between encapsulation, time, temperature, number of layers, and infill of the printed cookies was observed on the moisture and antioxidant activity. Four-layer cookies with 30% infill provided the highest bioactivity and phenolic content if baked for 10 min and at 180 °C. The bioacitivity and total phenolic content improved by 115% and 173%, respectively, comparing to free extract cookies.Moreover, the proper combination of the design and baking variables allowed to vary the bioactivity of cooked cookies (moisture 35%) between 300 and 700 ?molTR/gdry. The additive manufacture of foods with interconnected pores could accelerate baking and browning, or reduce thermal degradation. This represents a potential approach to enhance the functional and healthy properties of cookies or other thermal treated bioactive food products.The research leading to these results has received funding from FODIAC – Food for Diabetes and Cognition, funded by European Union, under the call Marie Skłodowsk-Curie Research and Innovation Staff Exchange (Ref. H2020-MSCA-RISE-778388); PhD grantship from Fondazione di Piacenza e Vigevano (Doctoral School on the Agro-Food System, Università Cattolica del Sacro Cuore); Fondazione Cariplo through the project ReMarcForFood – Biotechnological strategies for the conversion of Winemaking by-products and their recycling into the food chain: development of new concepts of use, 2016-0740 grant.info:eu-repo/semantics/publishedVersio

Bats, Trypanosomes, and Triatomines in Ecuador: New Insights into the Diversity, Transmission, and Origins of Trypanosoma cruzi and Chagas Disease

The generalist parasite Trypanosoma cruzi has two phylogenetic lineages associated almost exclusively with bats—Trypanosoma cruzi Tcbat and the subspecies T. c. marinkellei. We present new information on the genetic variation, geographic distribution, host associations, and potential vectors of these lineages. We conducted field surveys of bats and triatomines in southern Ecuador, a country endemic for Chagas disease, and screened for trypanosomes by microscopy and PCR. We identified parasites at species and genotype levels through phylogenetic approaches based on 18S ribosomal RNA (18S rRNA) and cytochrome b (cytb) genes and conducted a comparison of nucleotide diversity of the cytb gene. We document for the first time T. cruzi Tcbat and T. c. marinkellei in Ecuador, expanding their distribution in South America to the western side of the Andes. In addition, we found the triatomines Cavernicola pilosa and Triatoma dispar sharing shelters with bats. The comparisons of nucleotide diversity revealed a higher diversity for T. c. marinkellei than any of the T. c. cruzi genotypes associated with Chagas disease. Findings from this study increased both the number of host species and known geographical ranges of both parasites and suggest potential vectors for these two trypanosomes associated with bats in rural areas of southern Ecuador. The higher nucleotide diversity of T. c. marinkellei supports a long evolutionary relationship between T. cruzi and bats, implying that bats are the original hosts of this important parasite

Endosymbiosis in trypanosomatids: the genomic cooperation between bacterium and host in the synthesis of essential amino acids is heavily influenced by multiple horizontal gene transfers

Background Trypanosomatids of the genera Angomonas and Strigomonas live in a mutualistic association characterized by extensive metabolic cooperation with obligate endosymbiotic Betaproteobacteria. However, the role played by the symbiont has been more guessed by indirect means than evidenced. Symbiont-harboring trypanosomatids, in contrast to their counterparts lacking symbionts, exhibit lower nutritional requirements and are autotrophic for essential amino acids. To evidence the symbiont’s contributions to this autotrophy, entire genomes of symbionts and trypanosomatids with and without symbionts were sequenced here. Results Analyses of the essential amino acid pathways revealed that most biosynthetic routes are in the symbiont genome. By contrast, the host trypanosomatid genome contains fewer genes, about half of which originated from different bacterial groups, perhaps only one of which (ornithine cyclodeaminase, EC:4.3.1.12) derived from the symbiont. Nutritional, enzymatic, and genomic data were jointly analyzed to construct an integrated view of essential amino acid metabolism in symbiont-harboring trypanosomatids. This comprehensive analysis showed perfect concordance among all these data, and revealed that the symbiont contains genes for enzymes that complete essential biosynthetic routes for the host amino acid production, thus explaining the low requirement for these elements in symbiont-harboring trypanosomatids. Phylogenetic analyses show that the cooperation between symbionts and their hosts is complemented by multiple horizontal gene transfers, from bacterial lineages to trypanosomatids, that occurred several times in the course of their evolution. Transfers occur preferentially in parts of the pathways that are missing from other eukaryotes. Conclusion We have herein uncovered the genetic and evolutionary bases of essential amino acid biosynthesis in several trypanosomatids with and without endosymbionts, explaining and complementing decades of experimental results. We uncovered the remarkable plasticity in essential amino acid biosynthesis pathway evolution in these protozoans, demonstrating heavy influence of horizontal gene transfer events, from Bacteria to trypanosomatid nuclei, in the evolution of these pathways

Nonspecific Skin Reactivity in Epidemiological Studies

Os testes cutâneos por prick constituem o método diagnóstico de eleição no estudo da sensibilização alergénica, sendo no entanto influenciados por diversos factores. Não se encontram disponíveis dados sobre a quantificação da reacti-vidade cutânea inespecífica em estudos populacionais pediátricos, incluindo amostras de raças diversas. Objectivo: Avaliar e correlacionar a reactividade cutânea à histamina em crianças de diferentes raças,bem como a possível influência de factores como o sexo e a atopia. Material e Métodos: Englobado no Estudo Português de Doenças Alérgicas na Criança (PAC Study), foram estudadas 1710 crianças em idade escolar (6 a 12 anos), 244 de raça Negra na República de Cabo Verde, 756 Caucasianas na Ilha da Madeira e 710 de raça Chinesa em Macau. A todas as crianças foram efectuados testes cutâneos por prick com aeroalergenos, utilizando como referência positiva um extracto de histamina a 10mg/ml (Merck Allergopharma). Foi efectuada análise comparativa do diâmetro médio da pápula induzida pela histamina

Phylogenetic and syntenic data support a single horizontal transference to a Trypanosoma ancestor of a prokaryotic proline racemase implicated in parasite evasion from host defences

Abstract\ud \ud Background\ud Proline racemase (PRAC) enzymes of Trypanosoma cruzi (TcPRAC), the agent of Chagas disease, and Trypanosoma vivax (TvPRAC), the agent of livestock trypanosomosis, have been implicated in the B-cells polyclonal activation contributing to immunosuppression and the evasion of host defences. The similarity to prokaryotic PRAC and the absence in Trypanosoma brucei and Trypanosoma congolense have raised many questions about the origin, evolution, and functions of trypanosome PRAC (TryPRAC) enzymes.\ud \ud \ud Findings\ud We identified TryPRAC homologs as single copy genes per haploid genome in 12 of 15 Trypanosoma species, including T. cruzi and T. cruzi marinkellei, T. dionisii, T. erneyi, T. rangeli, T. conorhini and T. lewisi, all parasites of mammals. Polymorphisms in TcPRAC genes matched T. cruzi genotypes: TcI-TcIV and Tcbat have unique genes, while the hybrids TcV and TcVI contain TcPRACA and TcPRACB from parental TcII and TcIII, respectively. PRAC homologs were identified in trypanosomes from anurans, snakes, crocodiles, lizards, and birds. Most trypanosomes have intact PRAC genes. T. rangeli possesses only pseudogenes, maybe in the process of being lost. T. brucei, T. congolense and their allied species, except the more distantly related T. vivax, have completely lost PRAC genes.\ud \ud \ud \ud Conclusions\ud The genealogy of TryPRAC homologs supports an evolutionary history congruent with the Trypanosoma phylogeny. This finding, together with the synteny of PRAC loci, the relationships with prokaryotic PRAC inferred by taxon-rich phylogenetic analysis, and the absence in trypanosomatids of any other genera or in bodonids or euglenids suggest that a common ancestor of Trypanosoma gained PRAC gene by a single and ancient horizontal gene transfer (HGT) from a Firmicutes bacterium more closely related to Gemella and other species of Bacilli than to Clostridium as previously suggested. Our broad phylogenetic study allowed investigation of TryPRAC evolution over long and short timescales. TryPRAC genes diverged to become species-specific and genotype-specific for T. cruzi and T. rangeli, with resulting genealogies congruent with those obtained using vertically inherited genes. The inventory of TryPRAC genes described here is the first step toward the understanding of the roles of PRAC enzymes in trypanosomes differing in life cycles, virulence, and infection and immune evasion strategies.U.S. National Institutes of Health (NIH)U.S. National Science Foundation (NSF)SENACYT (Secretaría Nacional de Ciencia, Tecnología e Innovación)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)FAPESP (grant #2013/14622-3, São Paulo Research Foundation)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Universidade de São Paulo (USP

Phylogenetic and morphological characterization of trypanosomes from Brazilian armoured catfishes and leeches reveal high species diversity, mixed infections and a new fish trypanosome species

Abstract\ud \ud Background\ud Several Trypanosoma species transmitted by leeches infect marine and freshwater fish worldwide. To date, all South American fish trypanosome species identified have been based on unreliable morphological parameters. We recently isolated and cultured trypanosomes from the Brazilian armoured catfishes Hypostomus luetkeni and H. affinis. Here, we report the first phylogenetic analyses of South American (Brazilian) trypanosomes isolated from fish, and from leeches removed from these fish. We also analysed morphologically and morphometrically the different forms of fish, leech and cultured trypanosomes.\ud \ud \ud Methods\ud V7V8 SSU rRNA and gGAPDH sequences were used for phylogenetic analysis of Brazilian fish and leech trypanosomes. Trypanosomes from cultures, fish blood and leech samples were also characterized morphologically and morphometrically by light and electron microscopy.\ud \ud \ud Results\ud In blood smears from fish high trypanosome prevalence (90–100 %) and parasitemia (0.9-1.0x102) were observed. Phylogenetic relationships using SSU rRNA and gGAPDH showed that, despite relevant sequence divergence, all Brazilian fish (and derived cultures) and leech trypanosomes clustered together into a single clade. The Brazilian clade clustered with European, North American and African fish trypanosomes. Based on sequence analysis, we uncovered a new species of Brazilian fish trypanosome, Trypanosoma abeli n. sp. Trypanosoma abeli cultures contained pleomorphic epimastigotes, small trypomastigotes and rare sphaeromastigotes. Ultrastructural features of T. abeli included a cytostome-cytopharynx complex in epi- and trypomastigotes, a compact rod-like kinetoplast, lysosome-related organelles (LROs) and multivesicular bodies. Trypanosomes found in fish blood smears and leech samples were highly pleomorphic, in agreement with sequence data suggesting that catfishes and leeches often have mixed trypanosome infections.\ud \ud \ud Conclusions\ud \ud Trypanosoma abeli n. sp. is the first trypanosome from South American fishes isolated in culture, positioned in phylogenetic trees and characterized at the ultrastructural level. Trypanosoma abeli n. sp. is highly prevalent in H. luetkeni and H. affinis armoured catfish from the Atlantic Forest biome, and in other catfish species from the Amazon and the Pantanal. Sequencing data suggested that Brazilian catfish often have mixed trypanosome infections, highlighting the importance of molecular characterization to identify trypanosome species in fishes and leeches.We are grateful to Mr. Alcir Pereira de Souza, Ms. Alessandra Simões de\ud Toledo Pereira and Mrs. Tereza da Silva Lemos for their valuable support\ud with the fieldwork, and to Dr. José Carlos Oliveira and Valter M. AzevedoSantos\ud for fish identification. DNA sequencing and phylogenetic analyses\ud done in USP were supported by grants from CNPq and CAPES to MMGT. ML\ud is a postdoctoral fellow funded by CNPq, BRF is a PhD student funded by\ud CAPES, CSR is PhD student funded by CNPq, and LH is postdoctoral fellow\ud funded by FAPERJ

Genetic diversity and phylogenetic relationships of coevolving symbiont-harboring insect trypanosomatids, and their neotropical dispersal by invader African blowflies (Calliphoridae)

This study is about the inter- and intra-specific genetic diversity of trypanosomatids of the genus Angomonas, and their association with Calliphoridae (blowflies) in Neotropical and Afrotropical regions. Microscopic examination of 3,900 flies of various families, mostly Calliphoridae, revealed that 31% of them harbored trypanosomatids. Small subunit rRNA (SSU rRNA) barcoding showed that Angomonas predominated (46%) over the other common trypanosomatids of blowflies of genera Herpetomonas and Wallacemonas. Among Angomonas spp., A. deanei was much more common than the two-other species, A. desouzai and A. ambiguus. Phylogenetic analyses based on SSU rRNA, glycosomal glyceraldehyde-3-phosphate dehydrogenase (gGAPDH) and internal transcribed spacer rDNA (ITS rDNA) sequences revealed a marked genetic diversity within A. deanei, which comprised four infraspecific genotypes (Dea1– Dea4), and four corresponding symbiont genotypes (Kcr1–Kcr4). Host and symbiont phylogenies were highly congruent corroborating their co-divergence, consistent with host-symbiont interdependent metabolism and symbiont reduced genomes shaped by a long coevolutionary history. We compared the diversity of Angomonas/symbionts from three genera of blowflies, Lucilia, Chrysomya and Cochliomyia. A. deanei, A. desouzai, and A. ambiguus were found in the three genera of blowflies in South America. In Africa, A. deanei and A. ambiguus were identified in Chrysomya. The absence of A. desouzai in Africa and its presence in Neotropical Cochliomyia and Lucilia suggests parasite spillback of A. desouzai into Chrysomya, which was most likely introduced four decades ago from Africa into the Neotropic. The absence of correlation between parasite diversity and geographic and genetic distances, with identical genotypes of A. deanei found in the Neotropic and Afrotropic, is consistent with disjunct distribution due to the recent human-mediated transoceanic dispersal of Angomonas by Chrysomya. This study provides the most comprehensive data gathered so far on the genetic repertoires of a genus of trypanosomatids found in flies from a wide geographical range.The PROAFRICA, INCT-EPIAMO, and PROSUL programs of CNPq, PNIPB of Capes, and FAPESP (Process 2016/07487-0). CAPEs (PNPD) granted a postdoctoral scholarship to TB.http://www.frontiersin.org/Microbiologyam2018Veterinary Tropical Disease

Biosynthesis of vitamins and cofactors in bacterium-harbouring trypanosomatids depends on the symbiotic association as revealed by genomic analyses

International audienceSome non-pathogenic trypanosomatids maintain a mutualistic relationship with a betaproteobacterium of the Alcaligenaceae family. Intensive nutritional exchanges have been reported between the two partners, indicating that these protozoa are excellent biological models to study metabolic co-evolution. We previously sequenced and herein investigate the entire genomes of five trypanosomatids which harbor a symbiotic bacterium (SHTs for Symbiont-Haboring Trypanosomatids) and the respective bacteria (TPEs for Trypanosomatid Proteobacterial Endosymbiont), as well as two trypanosomatids without symbionts (RTs for Regular Trypanosomatids), for the presence of genes of the classical pathways for vitamin biosynthesis. Our data show that genes for the biosynthetic pathways of thiamine, biotin, and nicotinic acid are absent from all trypanosomatid genomes. This is in agreement with the absolute growth requirement for these vitamins in all protozoa of the family. Also absent from the genomes of RTs are the genes for the synthesis of pantothenic acid, folic acid, riboflavin, and vitamin B6. This is also in agreement with the available data showing that RTs are auxotrophic for these essential vitamins. On the other hand, SHTs are autotrophic for such vitamins. Indeed, all the genes of the corresponding biosynthetic pathways were identified, most of them in the symbiont genomes, while a few genes, mostly of eukaryotic origin, were found in the host genomes. The only exceptions to the latter are: the gene coding for the enzyme ketopantoate reductase (EC:1.1.1.169) which is related instead to the Firmicutes bacteria; and two other genes, one involved in the salvage pathway of pantothenic acid and the other in the synthesis of ubiquinone, that are related to Gammaproteobacteria. Their presence in trypanosomatids may result from lateral gene transfer. Taken together, our results reinforce the idea that the low nutritional requirement of SHTs is associated with the presence of the symbiotic bacterium, which contains most genes for vitamin production

Uses of innovative modeling tools within the implementation of the marine strategy framework directive

© 2016 Lynam, Uusitalo, Patrício, Piroddi, Queirós, Teixeira, Rossberg, Sagarminaga, Hyder, Niquil, Möllmann, Wilson, Chust, Galparsoro, Forster, Veríssimo, Tedesco, Revilla and Neville. In Europe and around the world, the approach to management of the marine environment has developed from the management of single issues (e.g., species and/or pressures) toward holistic Ecosystem Based Management (EBM) that includes aims to maintain biological diversity and protect ecosystem functioning. Within the European Union, this approach is implemented through the Marine Strategy Framework Directive (MSFD, 2008/56/EC). Integrated Ecosystem Assessment is required by the Directive in order to assess Good Environmental Status (GES). Ecological modeling has a key role to play within the implementation of the MSFD, as demonstrated here by case studies covering a range of spatial scales and a selection of anthropogenic threats. Modeling studies have a strong role to play in embedding data collected at limited points within a larger spatial and temporal scale, thus enabling assessments of pelagic and seabed habitat. Furthermore, integrative studies using food web and ecosystem models are able to investigate changes in food web functioning and biological diversity in response to changes in the environment and human pressures. Modeling should be used to: support the development and selection of specific indicators; set reference points to assess state and the achievement of GES; inform adaptive monitoring programs and trial management scenarios. The modus operandi proposed shows how ecological modeling could support the decision making process leading to appropriate management measures and inform new policy