Chitosan/PVA Based Membranes Processed by Gamma Radiation as Scaffolding Materials for Skin Regeneration

Some of the current strategies for the development of scaffolding materials capable of inducing tissue regeneration have been based on the use of polymeric biomaterials. Chitosan, in particular, due to its recognized biological activity has been used in a number of biomedical applications. Aiming the development of chitosan-based membranes with improved cell adhesion and growth properties to be used as skin scaffolds allowing functional tissue replacement, different formulations with chitosan of different molecular weight, poly (vinyl alcohol) and gelatin, were evaluated. To meet the goal of getting ready-to-use scaffolds assuring membranes’ required properties and sterilization, preparation methodology included a lyophilization procedure followed by a final gamma irradiation step. Two radiation dose values were tested. Samples were characterized by TGA, FTIR, and SEM techniques. Their hydrophilic properties, in vitro stability, and biocompatibility were also evaluated. Results show that all membranes present a sponge-type inner structure. Chitosan of low molecular weight and the introduction of gelatin are more favorable to cellular growth leading to an improvement on cells’ morphology and cytoskeletal organization, giving a good perspective to the use of these membranes as potential skin scaffolds.info:eu-repo/semantics/publishedVersio

Redefining the role of ectoderm in somitogenesis: a player in the formation of the fibronectin matrix of presomitic mesoderm

The absence of ectoderm impairs somite formation in cultured presomitic mesoderm (PSM) explants, suggesting that an ectoderm-derived signal is essential for somitogenesis. Here we show in chick that the standard enzymatic treatments used for explant isolation destroy the fibronectin matrix surrounding the anterior PSM, which fails to form somites when cultured for 6 hours. By contrast, explants isolated with collagenase retain their fibronectin matrix and form somites under identical culture conditions. The additional presence of ectoderm enhances somite formation, whereas endoderm has no effect. Furthermore, we show that pancreatin-isolated PSM explants cultured in fibronectin-supplemented medium, form significantly more somites than control explants. Interestingly, ectoderm is the major producer of fibronectin (Fn1) transcripts, whereas all but the anterior-most region of the PSM expresses the fibronectin assembly receptor, integrin alpha5 (Itga5). We thus propose that the ectoderm-derived fibronectin is assembled by mesodermal alpha5beta1 integrin on the surface of the PSM. Finally, we demonstrate that inhibition of fibronectin fibrillogenesis in explants with ectoderm abrogates somitogenesis. We conclude that a fibronectin matrix is essential for morphological somite formation and that a major, previously unrecognised role of ectoderm in somitogenesis is the synthesis of fibronectin.Fundação para a Ciência e a Tecnologia (FCT)/FEDER projects POCTI/BCI/40754/2001 and POCI/BIA-BCM/59201/2004 and the FP6/EU Network of Excellence ‘Cells into Organs’ of which P.R., C.L., R.P.A., G.R., I.P. and S.T. are members. L.C. was supported by the European Social Fund contract 1/3.2/PRODEP/2001. R.P.A. was supported by FCT (SFRH/BPD/9432/2002)

Chitosan-Based Membranes for Skin Wound Repair in a Dorsal Fold Chamber Rat Model

Frequently, deep partial and full-thickness skin wounds do not spontaneously regenerate. To restore the normal function of skin, epidermal and dermal components have to be supplied to the wound bed by grafting various substrates. Available options are limited and frequently costly. Herein, authors present a possible approach using 3D skin scaffolds capable of mimicking structure and biological functions of the extracellular matrix, providing, in parallel, a good environment for cell attachment, proliferation and differentiation. Low-molecular weight chitosan-based membranes were prepared by freeze-drying and ionizing radiation techniques to be used as skin scaffolds. Poly (vinyl alcohol), PVA, vinyl pyrrolidone, VP, and gelatin from cold water fish were incorporated. Information regarding membranes’ physical-chemical properties from SEM analysis, swelling and weight loss, together with biological response through in vitro assays (using Human Caucasian Fetal Foreskin Fibroblast) allowed the selection of an optimized batch of membranes that was used as skin scaffold in a dorsal rat model wound. The in vivo implantation assays (in Wistar rats) resulted in very promising results: (i) healing process faster than control; (ii) good vascularization; (iii) viable new tissues morphologically functional.info:eu-repo/semantics/publishedVersio

Standardization of F-18 by digital b-(LS)-g coincidence counting

The nuclide 18F disintegrates to 18O by β + emission (96.86%) and electron capture (3.14%) with a half-life of 1.8288 h. It is widely used in nuclear medicine for positron emission tomography (PET). Because of its short half-life this nuclide requires the development of fast measuring methods to be standardized. The combination of LSC methods with digital techniques proves to be a good alternative to get low uncertainties for this, and other, short lived nuclides. A radioactive solution of 18F has been standardized by coincidence counting with a LSC, using the logical sum of double coincidences in a TDCR array and a NaI scintillation detector. The results show good consistency with other techniques like 4πγ and LSC.Fil: Rodrigues Ferreira Maltez, Dario Pablo. Comision Nacional de Energia Atomica. Gerencia de Area de Aplicaciones de la Tecnología Nuclear. Gerencia D/aplicaciones y Tec de Radioisotopos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Balpardo, Christian Guillermo. Comision Nacional de Energia Atomica. Gerencia de Area de Aplicaciones de la Tecnología Nuclear. Gerencia D/aplicaciones y Tec de Radioisotopos; ArgentinaFil: Cassette, P.. Laboratoire National Henri Becquerel; FranciaFil: Arenillas, Pablo. Comision Nacional de Energia Atomica. Gerencia de Area de Aplicaciones de la Tecnología Nuclear. Gerencia D/aplicaciones y Tec de Radioisotopos; ArgentinaFil: Capoulat, Maria Eugenia. Comision Nacional de Energia Atomica. Gerencia de Area de Aplicaciones de la Tecnología Nuclear. Gerencia D/aplicaciones y Tec de Radioisotopos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Ceruti, Gabriela. Comision Nacional de Energia Atomica. Gerencia de Area de Aplicaciones de la Tecnología Nuclear. Gerencia D/aplicaciones y Tec de Radioisotopos; ArgentinaFil: Garcia Toraño, Eduado. Laboratorio de Metrología de Radiaciones Ionizantes; Españ

Surface Physicochemical Properties At The Micro And Nano Length Scales: Role On Bacterial Adhesion And Xylella Fastidiosa Biofilm Development.

The phytopathogen Xylella fastidiosa grows as a biofilm causing vascular occlusion and consequently nutrient and water stress in different plant hosts by adhesion on xylem vessel surfaces composed of cellulose, hemicellulose, pectin and proteins. Understanding the factors which influence bacterial adhesion and biofilm development is a key issue in identifying mechanisms for preventing biofilm formation in infected plants. In this study, we show that X. fastidiosa biofilm development and architecture correlate well with physicochemical surface properties after interaction with the culture medium. Different biotic and abiotic substrates such as silicon (Si) and derivatized cellulose films were studied. Both biofilms and substrates were characterized at the micro- and nanoscale, which corresponds to the actual bacterial cell and membrane/ protein length scales, respectively. Our experimental results clearly indicate that the presence of surfaces with different chemical composition affect X. fastidiosa behavior from the point of view of gene expression and adhesion functionality. Bacterial adhesion is facilitated on more hydrophilic surfaces with higher surface potentials; XadA1 adhesin reveals different strengths of interaction on these surfaces. Nonetheless, despite different architectural biofilm geometries and rates of development, the colonization process occurs on all investigated surfaces. Our results univocally support the hypothesis that different adhesion mechanisms are active along the biofilm life cycle representing an adaptation mechanism for variations on the specific xylem vessel composition, which the bacterium encounters within the infected plant.8e7524

Review of Piezoelectrical Materials Potentially Useful for Peripheral Nerve Repair

Funding Information: This research was funded by the International Atomic Energy Agency: Research Contract No. 24415 (CRP F23035). Authors acknowledge FCT—Fundação para a Ciência e a Tecnologia I.P. for the national funds in the scope of the project UIDB/04349/2020+UIDP/04349/2020, UIDB/00100/2020+LA/P/0056/2020 and UIDB/00329/2020. Publisher Copyright: © 2023 by the authors.It has increasingly been recognized that electrical currents play a pivotal role in cell migration and tissue repair, in a process named “galvanotaxis”. In this review, we summarize the current evidence supporting the potential benefits of electric stimulation (ES) in the physiology of peripheral nerve repair (PNR). Moreover, we discuss the potential of piezoelectric materials in this context. The use of these materials has deserved great attention, as the movement of the body or of the external environment can be used to power internally the electrical properties of devices used for providing ES or acting as sensory receptors in artificial skin (e-skin). The fact that organic materials sustain spontaneous degradation inside the body means their piezoelectric effect is limited in duration. In the case of PNR, this is not necessarily problematic, as ES is only required during the regeneration period. Arguably, piezoelectric materials have the potential to revolutionize PNR with new biomedical devices that range from scaffolds and nerve-guiding conduits to sensory or efferent components of e-skin. However, much remains to be learned regarding piezoelectric materials, their use in manufacturing of biomedical devices, and their sterilization process, to fine-tune their safe, effective, and predictable in vivo application.publishersversionpublishe

Novel scFv against Notch Ligand JAG1 Suitable for Development of Cell Therapies toward JAG1-Positive Tumors

Funding Information: This research was funded by the Fundação para a Ciência e Tecnologia/Ministério da Ciência, Tecnologia e Ensino Superior (FCT/MCTES, Portugal) grant PTDC/BBB-BMD/4497/2014 (to A.B.), through national funds to iNOVA4Health (UIDB/04462/2020 and UIDP/04462/2020), and the Associate Laboratory LS4FUTURE (LA/P/0087/2020). Publisher Copyright: © 2023 by the authors.The Notch signaling ligand JAG1 is overexpressed in various aggressive tumors and is associated with poor clinical prognosis. Hence, therapies targeting oncogenic JAG1 hold great potential for the treatment of certain tumors. Here, we report the identification of specific anti-JAG1 single-chain variable fragments (scFvs), one of them endowing chimeric antigen receptor (CAR) T cells with cytotoxicity against JAG1-positive cells. Anti-JAG1 scFvs were identified from human phage display libraries, reformatted into full-length monoclonal antibodies (Abs), and produced in mammalian cells. The characterization of these Abs identified two specific anti-JAG1 Abs (J1.B5 and J1.F1) with nanomolar affinities. Cloning the respective scFv sequences in our second- and third-generation CAR backbones resulted in six anti-JAG1 CAR constructs, which were screened for JAG1-mediated T-cell activation in Jurkat T cells in coculture assays with JAG1-positive cell lines. Studies in primary T cells demonstrated that one CAR harboring the J1.B5 scFv significantly induced effective T-cell activation in the presence of JAG1-positive, but not in JAG1-knockout, cancer cells, and enabled specific killing of JAG1-positive cells. Thus, this new anti-JAG1 scFv represents a promising candidate for the development of cell therapies against JAG1-positive tumors.publishersversionpublishe

Vitamin D3 as adjuvant in the treatment of type 2 diabetes mellitus: modulation of genomic and biochemical instability

Erratum in - Corrigendum: Vitamin D3 as adjuvant in the treatment of type 2 diabetes mellitus: modulation of genomic and biochemical instability. Fagundes GE, Macan TP, Rohr P, Damiani AP, Da Rocha FR, Pereira M, Longaretti LM, Vilela TC, Ceretta LB, Mendes C, Silveira PCL, Teixeira JPF, de Andrade VM. Mutagenesis. 2019 May 29;34(2):215. doi: 10.1093/mutage/gez006.Type 2 diabetes mellitus has undergone a worldwide growth in incidence in the world and has now acquired epidemic status. There is a strong link between type 2 diabetes and vitamin D deficiency. Because vitamin D has beneficial effects on glucose homeostasis, the aim of this study was to evaluate the influence of vitamin D3 supplementation on the modulation of glycaemic control and other metabolic effects, as well as modulation of genomic instability in patients with type 2 diabetes. We evaluated 75 patients with type 2 diabetes, registered in the Integrated Clinics of the University of Southern Santa Catarina. Participants received 4000 IU of vitamin D3 (25(OH)D) supplementation daily for 8 weeks. Blood samples were collected at the beginning and at the end of the supplementation, and 4 weeks after the end of supplementation. The glycidic and lipid profiles [total cholesterol, high-density lipoprotein (HDL), low-density lipoprotein and triglycerides], oxidative stress, DNA damage and 25(OH)D levels were evaluated. Vitamin D3 supplementation for 8 weeks showed enough to significantly increase blood levels of 25(OH)D. A significant difference in lipid profile was observed only in non-HDL cholesterol. Significant changes were observed in glucose homeostasis (fasting glucose and serum insulin) and, in addition, a reduction in the parameters of oxidative stress and DNA damage. There was a significant reduction in the values of 25(OH)D 4 weeks after the end of the supplementation, but levels still remained above baseline. Use of vitamin D supplementation can be an ally in the health modulation of patients with type 2 diabetes mellitusThis work was supported by grants from Conselho Nacional de Pesquisa e Desenvolvimento (CNPq), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Fundação de Amparo à Pesquisa e Inovação do Estado de Santa Catarina (FAPESC) and Programa de Pós-Graduação em Ciências da Saúde/ Universidade do Extremo Sul Catarinense.info:eu-repo/semantics/publishedVersio

Comparative genomic analysis of Leishmania (Viannia) peruviana and Leishmania (Viannia) braziliensis

BACKGROUND: The Leishmania (Viannia) braziliensis complex is responsible for most cases of New World tegumentary leishmaniasis. This complex includes two closely related species but with different geographic distribution and disease phenotypes, L. (V.) peruviana and L. (V.) braziliensis. However, the genetic basis of these differences is not well understood and the status of L. (V.) peruviana as distinct species has been questioned by some. Here we sequenced the genomes of two L. (V.) peruviana isolates (LEM1537 and PAB-4377) using Illumina high throughput sequencing and performed comparative analyses against the L. (V.) braziliensis M2904 reference genome. Comparisons were focused on the detection of Single Nucleotide Polymorphisms (SNPs), insertions and deletions (INDELs), aneuploidy and gene copy number variations. RESULTS: We found 94,070 variants shared by both L. (V.) peruviana isolates (144,079 in PAB-4377 and 136,946 in LEM1537) against the L. (V.) braziliensis M2904 reference genome while only 26,853 variants separated both L. (V.) peruviana genomes. Analysis in coding sequences detected 26,750 SNPs and 1,513 indels shared by both L. (V.) peruviana isolates against L. (V.) braziliensis M2904 and revealed two L. (V.) braziliensis pseudogenes that are likely to have coding potential in L. (V.) peruviana. Chromosomal read density and allele frequency profiling showed a heterogeneous pattern of aneuploidy with an overall disomic tendency in both L. (V.) peruviana isolates, in contrast with a trisomic pattern in the L. (V.) braziliensis M2904 reference. Read depth analysis allowed us to detect more than 368 gene expansions and 14 expanded gene arrays in L. (V.) peruviana, and the likely absence of expanded amastin gene arrays. CONCLUSIONS: The greater numbers of interspecific SNP/indel differences between L. (V.) peruviana and L. (V.) braziliensis and the presence of different gene and chromosome copy number variations support the classification of both organisms as closely related but distinct species. The extensive nucleotide polymorphisms and differences in gene and chromosome copy numbers in L. (V.) peruviana suggests the possibility that these may contribute to some of the unique features of its biology, including a lower pathology and lack of mucosal development. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12864-015-1928-z) contains supplementary material, which is available to authorized users

METHODS AIMED AT REDUCING THE RESIDUAL RISK OF PATHOGEN TRANSMISSION DURING PLATELET TRANSFUSION: A LITERATURE REVIEW

Objective: In the Brazilian public health system, sterile filtration is often used to prevent infections associated with platelet transfusion. The literature addressing this issue indicates that pathogen inactivation could be incorporated as an alternative to the development of individual tests and blood donor selection. The objective was to assess whether the use of platelets submitted to pathogen inactivation by photochemical methods could decrease the incidence of post-transfusion infections by viruses, bacteria or other pathogens compared to the use of platelet concentrate or platelets extracted by apheresis without photochemical treatment.Methods: A literature review from 1998 to 2015 was conducted. The scientific literature was surveyed using six electronic databases, two Internet search tools and a manual search of references, using specific search strategies for each database. The selected studies were assessed for quality according to a specific methodology. Data analysis was performed by observations made from the efficacy of the methods.Results: From a detailed analysis of 426 articles retrieved, 10 articles were selected for this review. Among the selected studies, seven studies were clinical trials, and three studies were systematic reviews in combination with meta-analysis. The outcomes analyzed included the reduction of the residual risk in pathogen transmission, mortality, occurrence of hemorrhagic events, corrected count increment (CCI) after 1 h, CCI after 24 h, and transfusion reactions.Conclusion: Differences were found in the quality of the included studies. Systematic reviews conducted on this topic, in alliance with political, social and administrative factors, will aid decision makers regarding its incorporation into the Brazilian Health System.Keywords: Blood Platelets, Platelet Transfusion, Platelet Transfusion/methods*, Riboflavin, Ultraviolet Rays, Photosensitizing Agents, Platelet Transfusion/adverse effect