Organogenic Nodule Formation in Hop: A Tool to Study Morphogenesis in Plants with Biotechnological and Medicinal Applications

The usage of Humulus lupulus for brewing increased the demand for high-quality plant material. Simultaneously, hop has been used in traditional medicine and recently recognized with anticancer and anti-infective properties. Tissue culture techniques have been reported for a wide range of species, and open the prospect for propagation of disease-free, genetically uniform and massive amounts of plants in vitro. Moreover, the development of large-scale culture methods using bioreactors enables the industrial production of secondary metabolites. Reliable and efficient tissue culture protocol for shoot regeneration through organogenic nodule formation was established for hop. The present review describes the histological, and biochemical changes occurring during this morphogenic process, together with an analysis of transcriptional and metabolic profiles. We also discuss the existence of common molecular factors among three different morphogenic processes: organogenic nodules and somatic embryogenesis, which strictly speaking depend exclusively on intrinsic developmental reprogramming, and legume nitrogen-fixing root nodules, which arises in response to symbiosis. The review of the key factors that participate in hop nodule organogenesis and the comparison with other morphogenic processes may have merit as a study presenting recent advances in complex molecular networks occurring during morphogenesis and together, these provide a rich framework for biotechnology applications

Assessment of stiffness and fatigue tests in Portugal

In the last decade, quality control of asphalt mixtures for pavement construction on the main Portuguese national road network used, most of the time, four-point bending beam tests (4pb) results as a reference. Stiffness modulus and fatigue laws established with samples prepared in the laboratory are usually used as a reference for behaviour/quality control analysis of the samples coming from the construction site. There is however, with this procedure, not only a problem of real world representation but also a question of the compatibility of results coming, again most of the time, from different laboratories using different equipments. Trying to address the second problem, this paper presents the results obtained from four-point bending tests carried out in three different laboratories in Portugal, owning three different 4pb equip-ments, concerning the performance characterization of a typical Portuguese base course asphalt mixture. The paper finally discusses the variability of the results obtained and underlines the main inferences that could be extracted

UHPLC-DAD-MSn analysis of phenolic compounds bioavailability throughout in vitro simulated gastrointestinal digestion

Phenolic compounds, one of the most widely distributed and structural diverse plant secondary metabolites families, have been the focus of several studies due to their vast range of biological activities (such as antioxidant, anti-inflammatory and/or antiproliferative). Since they are commonly present in human diet, phenolic compounds could be responsible for human health beneficial effects, preventing some disorders. Notwithstanding, these health effects are strictly dependent on their bioavailability, which consists in the amount of each ingested compound that reaches the target tissue where it can have a promising biological effect. Therefore, compound’s structure, human enzymatic activity and gut microbiota are some of the numerous factors that influenced phenolic compounds bioavailability, and consequently their human health beneficial effects.In this vein, the bioavailability of phenolic compounds from different classes, particularly, flavonols (rutin), flavanones (naringenin and naringin), dihydrochalcones (phloretin) and tannin monomeric units (phloroglucinol), were evaluated in an in vitro simulated gastrointestinal digestion and further analyzed and quantified through ultra-high performance liquid chromatography with diode-array detection and coupled to electrospray ionization tandem mass spectrometry (UHPLC-DAD-MSn). Most of them showed a bioavailability >70% in intestinal digestion phase and seemed to be absorbed, reaching the systemic circulation. Thus, these results could be a future remark to evaluate the human health effects of promising phenolic compounds combination, or of plant-based extracts with a similar composition or even extracts enriched with them.info:eu-repo/semantics/publishedVersio

Development and optimization of a new MALDI-TOF protocol for identification of the Sporothrix species complex

Accurate species identification of the Sporothrix schenckii complex is essential, since identification based only on phenotypic characteristics is often inconclusive due to phenotypic variability within the species. We used matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) for species identification of 70 environmental and clinical isolates of the Sporothrix complex. A reference database was established for MALDI-TOF MS-based species identification according to minor adjustments in the manufacturer's guidelines. The MALDI-TOF MS clearly distinguished strains of Sporothrix brasiliensis, Sporothrix globosa, Sporothrix mexicana, S. schenckii, Sporothrix luriei and Sporothrix pallida, enabling identification of all isolates at the species level, as confirmed by partial calmodulin gene sequence analyses. The present methodology is simple, reliable, rapid and highly suitable for routine identification in clinical mycology laboratories and culture collections, particularly for updating and reclassifying of deposited Sporothrix isolates.The authors wish to thank the following international researchers for generously contributing strains to this study: Conchita Torrielo (EH194, EH252, EH253); Myrtha Arango (04015, 11029, 010221, 10036, 03017, 03022, 12013, 03003, 14879); and Masako Kawasaki (KMU975). Financial support was provided by FAPERJ/Rio de Janeiro, Brazil (grant proc. E-26/110.619/2012) and PAPES VI-Fiocruz/CNPq (Proc. 407693/2012-2) R. M. Z-O. is supported in part by CNPq 304976/2013-0 and FAPERJ E-26/103.157/2011. M. M. E. O. was supported by a grant from CAPES 2445/11-5 and PNPD/CAPES-Fiocruz/Pesquisa Clinica em Doencas Infecciosas. M. M. E. O., C. S. and N. L. thank the FCT Strategic Project PEst-OE/EQB/LA0023/2013 and BioHealth-Biotechnology and Bioengineering approaches to improve health quality, Ref. NORTE-07-0124-FEDER-000027" co-funded by the Programa Operacional Regional do Norte (ON.2 - O Novo Norte), QREN, FEDER. Automated sequencing was done using the genomic platform/DNA sequencing platform at the Fundacao Oswaldo Cruz-PDTIS/FIOCRUZ (RPT01A), Brazil

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

Evaluation of different analytical approaches using total reflection X-ray fluorescence systems for multielemental analysis of human tissues with different adipose content

Publisher Copyright: © 2022 The AuthorsElemental content plays an important role in biological processes, and so, the multielemental analysis of human tissue samples is required in biomedical research. Still, the small amount of available biological samples and the adipose content of the samples can be major setbacks for the accurate determination of elemental content. In this study, we explored the potential of several analytical approaches combined with total reflection X-ray fluorescence spectrometry (TXRF) for multielemental analysis of human tissues with different adipose content (colon, heart, liver, lung, muscle, intestine, skin, stomach, uterus, bladder and aorta). The capabilities and limitations of different sample treatment procedures (suspension and acidic digestion) and two TXRF systems with different anode configurations (Mo and W X-ray tubes) have been evaluated for such purpose. Results showed that for tissues with a higher fat content (e.g., skin, and intestine) the best strategy was the acidic digestion of the sample before TXRF analysis. However, for other tissues, acceptable results were obtained by suspending 20 mg of powdered material in 1 mL of 2 M nitric acid. A further enhancement of the limits of detection and accuracy of the results was achieved if using Mo-TXRF systems, especially for the determination of low Z elements (e.g., K, and Ca) and of elements present at low concentrations (e.g., Cu) in the human tissues. Finally, results by TXRF analysis were compared with those obtained with μ-EDXRF and ICP-OES, and a good agreement was obtained.publishersversionpublishe