Standing of nucleic acid testing strategies in veterinary diagnosis laboratories to uncover Mycobacterium tuberculosis complex members

Nucleic acid testing (NAT) designate any molecular approach used for the detection, identification, and characterization of pathogenic microorganisms, enabling the rapid, specific, and sensitive diagnostic of infectious diseases, such as tuberculosis. These assays have been widely used since the 90s of the last century in human clinical laboratories and, subsequently, also in veterinary diagnostics. Most NAT strategies are based in the polymerase chain reaction (PCR) and its several enhancements and variations. From the conventional PCR, real-time PCR and its combinations, isothermal DNA amplification, to the nanotechnologies, here we review how the NAT assays have been applied to decipher if and which member of the Mycobacterium tuberculosis complex is present in a clinical sample. Recent advances in DNA sequencing also brought new challenges and have made possible to generate rapidly and at a low cost, large amounts of sequence data. This revolution with the high-throughput sequencing (HTS) technologies makes whole genome sequencing (WGS) and metagenomics the trendiest NAT strategies, today. The ranking of NAT techniques in the field of clinical diagnostics is rising, and we provide a SWOT (Strengths, Weaknesses, Opportunities, and Threats) analysis with our view of the use of molecular diagnostics for detecting tuberculosis in veterinary laboratories, notwithstanding the gold standard being still the classical culture of the agent. The complementary use of both classical and molecular diagnostics approaches is recommended to speed the diagnostic, enabling a fast decision by competent authorities and rapid tackling of the disease.publishersversionpublishe

Erratum: Moreira, J., et al., Spin-Coated Polysaccharide-Based Multilayered Freestanding Films with Adhesive and Bioactive Moieties. Molecules 2020, 25, 840

Erratum: Moreira, J., et al., Spin-Coated Polysaccharide-Based Multilayered Freestanding Films with Adhesive and Bioactive Moieties. Molecules 2020, 25, 840. DOI: 10.3390/molecules25040840The authors wish to make changes to the published paper 11 j. 1. UV-Vis Analysis of Catechol-Modified Polymers In the original manuscript theie is a mistake concerning the word "Wavenumber" in the X-Coordinate in Figure 1. Tile corrected word is "Wavelength". Tlx- A uthors also wish to change mg«mL-l to mg ml-1 in the legend of Figure l;see corrected Figure 1 below. (Figure Presented).(undefined

Chemical Modification of Semiconductor Surfaces by Means of Nanometric Cellulose Films

Cellulose films of nanometric thickness were produced by spin-coating on GaAs(100). Films were deposited using cellulose silylated solutions and subsequently regenerated by exposing them to vapors of hydrochloric acid. After regeneration, these films can strongly resist solvents. Modification of the film surface region was performed by immersing the regenerated cellulose films in a solution of phenyl isocyanate in dimethyl sulfoxide. A different functionalization was also successfully achieved through the interaction of the film surface with 4,4′-methylenebis(phenyl isocyanate) (MDI). Surfaces treated with MDI keep an unreacted isocyanate group and can again be modified by amines. For this purpose, 4-bromoaniline was used. All kinetics of the different molecular interactions with the cellulose film on GaAs were followed in situ using FTIRS in ATR/MIR (attenuated total reflection in multiple internal reflections) mode. Besides ATR/MIR having an analysis depth on the order of 1 µm, other surface techniques were used for analyzing these films with other probing depths such as X-ray photoelectron spectroscopy with ∼10 nm and high-resolution electron energy loss spectroscopy with ∼1 nm in the impact regime. The set of methods presented here represents a quite adequate way to study the surface chemistry of cellulose films and the procedures for their functionalization

Magnetic-responsive hydrogels for cartilage tissue engineering

Publicado em "Journal of Tissue Engineering and Regenerative Medicine", vol. 7, supp. 1 (2013)The use of magnetic nanoparticles (MNPs) has been explored as an alternative approach to overcome current limitations of regenerative medicine strategies. Cell engineering approaches where MNPs are incorporated within three-dimensional constructs, such as scaffolds or hydrogels may constitute a novel and attractive approach towards the development of a magnetically-responsive system. These systems would enable remote controlled actions over tissue engineered constructs in vitro and in vivo. Moreover, growing evidence suggests that the application of a magnetic field may enhance biological performance over commonly used static culture conditions providing stimulation for cell proliferation, migration and differentiation. In this work we analyze the role of magnetic stimulation on the behavior of human adipose derived stem cells (hASCs) laden in k-carrageenan hydrogels aiming at cartilage tissue engineering approaches. Thermo-responsive natural-based κ-carrageenan hydrogels were used as 3D templates since previous studies(1) report the adequate environment provided by these materials to support the viability and chondrogenic differentiation of several types of cells

Effects of myenteric denervation on extracellular matrix fibers and mast cell distribution in normal stomach and gastric lesions

Abstract\ud \ud \ud \ud Background\ud \ud In this study the effect of myenteric denervation induced by benzalconium chloride (BAC) on distribution of fibrillar components of extracellular matrix (ECM) and inflammatory cells was investigated in gastric carcinogenesis induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). Rats were divided in four experimental groups: non-denervated (I) and denervated stomach (II) without MNNG treatment; non-denervated (III) and denervated stomachs (IV) treated with MNNG. For histopathological, histochemical and stereological analysis, sections of gastric fragments were stained with Hematoxylin-Eosin, Picrosirius-Hematoxylin, Gomori reticulin, Weigert's Resorcin-Fuchsin, Toluidine Blue and Alcian-Blue/Safranin (AB-SAF).\ud \ud \ud \ud Results\ud \ud BAC denervation causes an increase in the frequency of reticular and elastic fibers in the denervated (group II) compared to the non-denervated stomachs (group I). The treatment of the animals with MNNG induced the development of adenocarcinomas in non-denervated and denervated stomachs (groups III and IV, respectively) with a notable increase in the relative volume of the stroma, the frequency of reticular fibers and the inflammatory infiltrate that was more intense in group IV. An increase in the frequency of elastic fibers was observed in adenocarcinomas of denervated (group IV) compared to the non-denervated stomachs (group III) that showed degradation of these fibers. The development of lesions (groups III and IV) was also associated with an increase in the mast cell population, especially AB and AB-SAF positives, the latter mainly in the denervated group IV.\ud \ud \ud \ud Conclusions\ud \ud The results show a strong association in the morphological alteration of the ECM fibrillar components, the increased density of mast cells and the development of tumors induced by MNNG in the non-denervated rat stomach or denervated by BAC. This suggests that the study of extracellular and intracellular components of tumor microenvironment contributes to understanding of tumor biology by action of myenteric denervation.We are grateful to Domingos Zanchetta Netto and Luiz Roberto Falleiros-Jr for technical assistance. CFE and CBM were supported by Fundação de Amparo á Pesquisa - FAPESP (grants 08/05722-6 and 03/10634-5, respectively) and SRT by Conselho Nacional de Desenvolvimento Cientifico e Tecnológico - CNPq (grants 301111/05-7 and 300163/2008-8).We are grateful to Domingos Zanchetta Netto and Luiz Roberto FalleirosJr for technical assistance. CFE and CBM were supported by Fundação de Amparo á Pesquisa FAPESP (grants 08/057226 and 03/106345, respectively) and SRT by Conselho Nacional de Desenvolvimento Cientifico e Tecnológico CNPq (grants 301111/057 and 300163/20088)

Staphylococcus aureus virulence factors and disease

Staphylococcus aureus is a major cause of nosocomial infections worldwide, especially methicillin-resistant S. aureus. Patients subjected to broad-spectrum antibiotics and immunosuppressive therapies have higher risk of infection by this microorganism. S. aureus infection are often extremely difficult to treat due to the large population heterogeneity, phenotypic switching, intra-strain diversity, hypermutability and most importantly the small colony variants. It is very important to emphasise that host immune responses against persistent infections by S. aureus is insufficient resulting normally into chronic infections, which in turn can lead to life threatening situations. So, throughout this chapter we will focus on the principal aspects of S. aureus virulence will be focused

Cold atmospheric plasma, a novel approach against bladder cancer, with higher sensitivity for the high-grade cell line

Antitumor therapies based on Cold Atmospheric Plasma (CAP) are an emerging medical field. In this work, we evaluated CAP effects on bladder cancer. Two bladder cancer cell lines were used, HT-1376 (stage III) and TCCSUP (stage IV). Cell proliferation assays were performed evaluating metabolic activity (MTT assay) and protein content (SRB assay). Cell viability, cell cycle, and mitochondrial membrane potential (Δψm) were assessed using flow cytometry. Reactive oxygen and nitrogen species (RONS) and reduced glutathione (GSH) were evaluated by fluorescence. The assays were carried out with different CAP exposure times. For both cell lines, we obtained a significant reduction in metabolic activity and protein content. There was a decrease in cell viability, as well as a cell cycle arrest in S phase. The Δψm was significantly reduced. There was an increase in superoxide and nitric oxide and a decrease in peroxide contents, while GSH content did not change. These results were dependent on the exposure time, with small differences for both cell lines, but overall, they were more pronounced in the TCCSUP cell line. CAP showed to have a promising antitumor effect on bladder cancer, with higher sensitivity for the high-grade cell line.info:eu-repo/semantics/publishedVersio

Fast calculation of spectral optical properties and pigment content detection in human normal and pathological kidney

A fast calculation method was used to obtain the spectral optical properties of human normal and pathological (chromophobe renal cell carcinoma) kidney tissues. Using total transmittance, total reflectance and collimated transmittance spectra acquired from ex vivo kidney samples, the spectral optical properties of both tissues, namely the absorption, the scattering and the reduced scattering coefficients, as well as the scattering anisotropy, dispersion and light penetration depth, were calculated between 200 and 1000 nm. Analysis of the mean ab sorption coefficient spectra of the kidney tissues showed that both contain melanin and lipofuscin, and that 83 % of the melanin in the normal kidney converts into lipofuscin in the pathological kidney.info:eu-repo/semantics/publishedVersio

Beyond the limits of oxygen: effects of hypoxia in a hormone-independent prostate cancer cell line

Prostate cancer (PCa) has a high incidence worldwide. One of the major causes of PCa resistance is intratumoral hypoxia. In solid tumors, hypoxia is strongly associated with malignant progression and resistance to therapy, which is an indicator of poor prognosis. The antiproliferative effect and induced death caused by doxorubicin, epirubicin, cisplatin, and flutamide in a hormone-independent PCa cell line will be evaluated. The hypoxia effect on drug resistance to these drugs, as well as cell proliferation and migration, will be also analyzed. All drugs induced an antiproliferative effect and also cell death in the cell line under study. Hypoxia made the cells more resistant to all drugs. Moreover, our results reveal that long time cell exposure to hypoxia decreases cellular proliferation and migration. Hypoxia can influence cellular resistance, proliferation, and migration. This study shows that hypoxia may be a key factor in the regulation of PCa.info:eu-repo/semantics/publishedVersio