Vibrio proteases for biomedical applications: Modulating the proteolytic secretome of v. alginolyticus and v. parahaemolyticus for improved enzymes production

Proteolytic enzymes are of great interest for biotechnological purposes, and their large-scale production, as well as the discovery of strains producing new molecules, is a relevant issue. Collagenases are employed for biomedical and pharmaceutical purposes. The high specificity of collagenase-based preparations toward the substrate strongly relies on the enzyme purity. However, the overall activity may depend on the cooperation with other proteases, the presence of which may be essential for the overall enzymatic activity, but potentially harmful for cells and tissues. Vibrios produce some of the most promising bacterial proteases (including collagenases), and their exo-proteome includes several enzymes with different substrate specificities, the production and relative abundances of which strongly depend on growth conditions. We evaluated the effects of different media compositions on the proteolytic exo-proteome of Vibrio alginolyticus and its closely relative Vibrio parahaemolyticus, in order to improve the overall proteases production, as well as the yield of the desired enzymes subset. Substantial biological responses were achieved with all media, which allowed defining culture conditions for targeted improvement of selected enzyme classes, besides giving insights in possible regulatory mechanisms. In particular, we focused our efforts on collagenases production, because of the growing biotechnological interest due to their pharmaceutical/biomedical applications

Teaching and learning Landscape Ecology to Landscape Architects in Italy : toward protective, adaptative, redundant landscape design

Some words are more and more used by different disciplines to focus on contemporary challenges, for exampe \u201csustainability\u201d and \u201cresilience\u201d, becoming trendy slogans, but the real understanding of these concepts in Landscape Ecology is necessary to avoid their loss of significance, and to add effectiveness to ecological based projects and actions. The collaborative partnership between Landscape Ecology and Landscape Architecture is a fundamental opportunity. Landscape Ecology is a necessary topic for landscape architects' education and its application becomes a tool for landscape projects. A good landscape architect can play a significative role in the promotion of people's appreciation of landscape in terms of resources (cfr. Almo Farina " theory of resources"). Starting from the experience, of more than 30 years, by the Genoese Landscape Architecture School, from the theoric teaching by Almo Farina and Vittorio Ingegnoli (see Ecofield theory, spatial configuration of functional elements , shifting mosaic, BTC measurement, Landscape Bionomics), to the applicative courses and experimental design of landscapes at different dimensions, the discipline of Landscape Ecology is a clear guide to the understanding of landscape configuration, and of its critical actual aspects. In the actual main Italian Schools of Landscape Architecture (Genoa/Turin/Milan, Florence, Rome, Milan) a fundamental role is given to Landscape Ecology education. Particularly in the Genoa/Turin/Milan Master Degree in Landscape Architecture, the experimental applicative approach to design by Landscape Ecology (in the brilliant courses of Applied Landscape Ecology by Gioia Gibelli and Luigino Pirola, with the help of Applied Botany), gives necessary tools to face the challenges of contemporary society, with particular reference to resources and needs, such as water, food production, ecosystemic services, socio-ecological relations, safetiness of everyday landscapes

Calibration of a Bending Model for Cold-formed Sections

The calibration procedure of a numerical model based on the simulation of the moment-curvature relation for cold-formed steel beams is presented. The results of bending tests on beams with double-channel cross-section are used to assess the proper values to the parameters which quantify the model

Native rodent species are unlikely sources of infection for Leishmania (Viannia) braziliensis along the Transoceanic Highway in Madre de Dios, Peru.

An estimated 2.3 million disability-adjusted life years are lost globally from leishmaniasis. In Peru's Amazon region, the department of Madre de Dios (MDD) rises above the rest of the country in terms of the annual incidence rates of human leishmaniasis. Leishmania (Viannia) braziliensis is the species most frequently responsible for the form of disease that results in tissue destruction of the nose and mouth. However, essentially nothing is known regarding the reservoirs of this vector-borne, zoonotic parasite in MDD. Wild rodents have been suspected, or proven, to be reservoirs of several Leishmania spp. in various ecosystems and countries. Additionally, people who live or work in forested terrain, especially those who are not regionally local and whose immune systems are thus naïve to the parasite, are at most risk for contracting L. (V.) braziliensis. Hence, the objective of this study was to collect tissues from wild rodents captured at several study sites along the Amazonian segment of the newly constructed Transoceanic Highway and to use molecular laboratory techniques to analyze samples for the presence of Leishmania parasites. Liver tissues were tested via polymerase chain reaction from a total of 217 rodents; bone marrow and skin biopsies (ear and tail) were also tested from a subset of these same animals. The most numerous rodent species captured and tested were Oligoryzomys microtis (40.7%), Hylaeamys perenensis (15.7%), and Proechimys spp. (12%). All samples were negative for Leishmania, implying that although incidental infections may occur, these abundant rodent species are unlikely to serve as primary reservoirs of L. (V.) braziliensis along the Transoceanic Highway in MDD. Therefore, although these rodent species may persist and even thrive in moderately altered landscapes, we did not find any evidence to suggest they pose a risk for L. (V.) braziliensis transmission to human inhabitants in this highly prevalent region

3d collagen hydrogel promotes in vitro langerhans islets vascularization through ad-mvfs angiogenic activity

Adipose derived microvascular fragments (ad-MVFs) consist of effective vascularization units able to reassemble into efficient microvascular networks. Because of their content in stem cells and related angiogenic activity, ad-MVFs represent an interesting tool for applications in regenerative medicine. Here we show that gentle dissociation of rat adipose tissue provides a mixture of ad-MVFs with a length distribution ranging from 33–955 µm that are able to maintain their original morphology. The isolated units of ad-MVFs that resulted were able to activate transcriptional switching toward angiogenesis, forming tubes, branches, and entire capillary networks when cultured in 3D collagen type-I hydrogel. The proper involvement of metalloproteases (MMP2/MMP9) and serine proteases in basal lamina and extracellular matrix ECM degradation during the angiogenesis were concurrently assessed by the evaluation of alpha-smooth muscle actin (αSMA) expression. These results suggest that collagen type-I hydrogel provides an adequate 3D environment supporting the activation of the vascularization process. As a proof of concept, we exploited 3D collagen hydrogel for the setting of ad-MVF–islet of Langerhans coculture to improve the islets vascularization. Our results suggest potential employment of the proposed in vitro system for regenerative medicine applications, such as the improving of the islet of Langerhans engraftment before transplantation