Testing the role of mate recognition proteins in a incipient ecological spreciation process

Comunicaciones a congreso

LYTAG-driven purification strategies as a key to integrate and intensify the downstream processing of monoclonal antibodies

Monoclonal antibodies (mAbs) are currently the most important class of recombinant protein therapeutics in the biotechnological and biopharmaceutical industry with more than 250 mAbs currently undergoing clinical trials. High titer producing cultures and complex mixtures containing high cell densities, together with an increasing growing demand for highly pure mAbs is making recovery and purification processes hot targets for improvement and opens important technological challenges in mAbs manufacturing platforms. This work explores the use of an affinity dual ligand based on a choline binding polypeptide tag (LYTAG) fused with the synthetic antibody Z domain (LYTAG-Z) as a tool to integrate and optimized the downstream processing of mAbs. Upon addition of this ligand to an animal cell culture broth, antibody-LYTAG-Z complexes are formed which can be easily captured and separated from host cell impurities by affinity partitioning in aqueous two-phase systems (ATPS) composed of polyethylene glycol –PEG, as PEG molecules have the ability to binding to the choline binding sites of LYTAG. Integration of clarification and primary mAbs recovery was successfully accomplished using a system composed of 6% PEG 3350 Da and 7% dextran 500,000 Da in which an extraction yield of 89% and a clarification higher than 95% were achieved. IgG-rich phases were further processed by chromatography, using three different strong anion exchange matrices charged with quaternary methyl amines (a choline analogue) – CIMmultus QA, HiTrap Q FF and gPore NW Q. A two-elution method was developed for the separation of the antibody-LYTAG-Z complexe, allowing simultaneous purification of the antibody and recovery of the ligand. The process was successfully scale-up 10000 times allowing a global antibody recovery of 70% with a purity of 89% and enabling 100% cell removal

Combustion Synthesis of Ultrafine Powders of Co3O4 for Selective Surfaces of Solar Collectors

Solar selective paints, with the addition of Co3O4 as a pigment, are used to improve energetic efficiency in solar collectors. Although Co3O4has been obtained by different methods, references about combustion synthesis are scarce. Co3O4 powders have been synthesized by stoichiometric and non-stoichiometric routes using aspartic acid (Asp) or tri-hydroxi-methyl-aminomethane (Tris) as fuels. The samples were calcined in air at 500 °C. They were characterized by X-ray diffraction, scanning electron microscopy, transmission electron microscopy, thermogravimetric analysis, differential scanning calorimetry, Fourier transform infrared spectrum and the specific surface area of the samples was determined by means of the Brunauer–Emmett–Teller technique. The optical properties of pigments were assessed by means of a spectrophotometer. In all cases, powders exhibited the crystalline structure of Co3O4. A minimum crystallite average size of 29 nm was observed for powders obtained by the “stoichiometric/Asp” combustion route, while a maximum value of 41 nm was stated for powders obtained by the “non-stoichiometric/Asp” combustion process. The average particle size ranged between 50 and 100 nm. The powders obtained by the“stoichiometric/Asp” method were selected to study their optical properties; their solar absorption value was 86%. Solar selective surfaces composed by Co3O4 pigments and an alkyd resin were obtained and applied over copper or aluminum substrates. In both cases, solar absorptance was of 93% and comparable with similar solar selective surfaces, but the thermal emittance value was higher than 90%, as a consequence of the large width of the films

Autocrine regulation of human sperm motility by tachykinins

<p>Abstract</p> <p>Background</p> <p>We examined the presence and function of tachykinins and the tachykinin-degrading enzymes neprilysin (NEP) and neprilysin-2 (NEP2) in human spermatozoa.</p> <p>Methods</p> <p>Freshly ejaculated semen was collected from forty-eight normozoospermic human donors. We analyzed the expression of substance P, neurokinin A, neurokinin B, hemokinin-1, NEP and NEP2 in sperm cells by reverse-transcriptase polymerase chain reaction (RT-PCR), western blot and immunocytochemistry assays and evaluated the effects of the neprilysin and neprilysin-2 inhibitor phosphoramidon on sperm motility in the absence and presence of tachykinin receptor-selective antagonists. Sperm motility was measured using WHO procedures or computer-assisted sperm analysis (CASA).</p> <p>Results</p> <p>The mRNAs of the genes that encode substance P/neurokinin A (TAC1), neurokinin B (TAC3), hemokinin-1 (TAC4), neprilysin (MME) and neprilysin-2 (MMEL1) were expressed in human sperm. Immunocytochemistry studies revealed that tachykinin and neprilysin proteins were present in spermatozoa and show specific and differential distributions. Phosphoramidon increased sperm progressive motility and its effects were reduced in the presence of the tachykinin receptor antagonists SR140333 (NK1 receptor-selective) and SR48968 (NK2 receptor-selective) but unmodified in the presence of SR142801 (NK3 receptor-selective).</p> <p>Conclusion</p> <p>These data show that tachykinins are present in human spermatozoa and participate in the regulation of sperm motility. Tachykinin activity is regulated, at least in part, by neprilysins.</p

Methodology for phytoplankton taxonomic group identification towards the development of a lab-on-a-chip

This paper presents the absorbance and fluorescence optical properties of various phytoplankton species, looking to achieve an accurate method to detect and identify a number of phytoplankton taxonomic groups. The methodology to select the excitation and detection wavelengths that results in superior identification of phytoplankton is reported. The macroscopic analyses and the implemented methodology are the base for designing a lab-on-a-chip device for a phytoplankton group identification, based on cell analysis with multi-wavelength lighting excitation, aiming for a cheap and portable platform. With such methodology in a lab-on-a-chip device, the analysis of the phytoplankton cells’ optical properties, e.g., fluorescence, diffraction, absorption and reflection, will be possible. This device will offer, in the future, a platform for continuous, autonomous and in situ underwater measurements, in opposition to the conventional methodology. A proof-of-concept device with LED light excitation at 450 nm and a detection photodiode at 680 nm was fabricated. This device was able to quantify the concentration of the phytoplankton chlorophyll a. A lock-in amplifier electronic circuit was developed and integrated in a portable and low-cost sensor, featuring continuous, autonomous and in situ underwater measurements. This device has a detection limit of 0.01 µ/L of chlorophyll a, in a range up to 300 µg/L, with a linear voltage output with chlorophyll concentration.Fundação para a Ciência e a Tecnologia | Ref. UIDB/04436/2020Fundação para a Ciência e a Tecnologia | Ref. UIDP/04436/2020Fundação para a Ciência e a Tecnologia | Ref. PD/BD/150581/2020Fundação para a Ciência e a Tecnologia | Ref. 2021.01087.CEECINDFundação para a Ciência e a Tecnologia | Ref. 2021.01086.CEECIN

The mechano-ubiquitinome of articular cartilage: differential ubiquitination and activation of a group of ER-associated DUBs and ER stress regulators

Understanding how connective tissue cells respond to mechanical stimulation is important to human health and disease processes in musculoskeletal diseases. Injury to articular cartilage is a key risk factor in predisposition to tissue damage and degenerative osteoarthritis. Recently, we have discovered that mechanical injury to connective tissues including murine and porcine articular cartilage causes a significant increase in Lysine 63- polyubiquitination. Here we identified the ubiquitin signature that is unique to injured articular cartilage tissue post mechanical injury (the “mechano-ubiquitinome”). A total of 463 ubiquitinated peptides were identified, with an enrichment of ubiquitinated peptides of proteins involved in protein processing in the endoplasmic reticulum (ER), also known as the ER-associated degradation (ERAD) response, including YOD1, BRCC3, ATXN3 and USP5 as well as the ER stress regulators, RAD23B, VCP/p97 and Ubiquilin 1. Enrichment of these proteins suggested an injury-induced ER stress response and, for instance, ER stress markers DDIT3/CHOP and BIP/GRP78 were upregulated following cartilage injury on the protein and gene expression levels. Similar ER stress induction was also observed in response to tail fin injury in zebrafish larvae, suggesting a generic response to tissue injury. Furthermore, a rapid increase in global DUB activity following injury and significant activity in human osteoarthritic cartilage was observed using DUB specific activity probes. Combined, these results implicate the involvement of ubiquitination events and activation of a set of DUBs and ER stress regulators in cellular responses to cartilage tissue injury and in osteoarthritic cartilage tissues. This link through the ERAD pathway makes this protein set attractive for further investigation in in vivo models of tissue injury and for targeting in osteoarthritis and related musculoskeletal diseases

Methodology for phytoplankton taxonomic group identification towards the development of a lab-on-a-chip

This paper presents the absorbance and fluorescence optical properties of various phytoplankton species, looking to achieve an accurate method to detect and identify a number of phytoplankton taxonomic groups. The methodology to select the excitation and detection wavelengths that results in superior identification of phytoplankton is reported. The macroscopic analyses and the implemented methodology are the base for designing a lab-on-a-chip device for a phytoplankton group identification, based on cell analysis with multi-wavelength lighting excitation, aiming for a cheap and portable platform. With such methodology in a lab-on-a-chip device, the analysis of the phytoplankton cells’ optical properties, e.g., fluorescence, diffraction, absorption and reflection, will be possible. This device will offer, in the future, a platform for continuous, autonomous and in situ underwater measurements, in opposition to the conventional methodology. A proof-of-concept device with LED light excitation at 450 nm and a detection photodiode at 680 nm was fabricated. This device was able to quantify the concentration of the phytoplankton chlorophyll a. A lock-in amplifier electronic circuit was developed and integrated in a portable and low-cost sensor, featuring continuous, autonomous and in situ underwater measurements. This device has a detection limit of 0.01 µ/L of chlorophyll a, in a range up to 300 µg/L, with a linear voltage output with chlorophyll concentration.European Regional Development Fund (ERDF) through the Interreg VA Spain-Portugal (POCTEP) 2014–2020 Program under grant agreement 0591_FOODSENS_1_E, under the national support to R&D units grant, through the reference project UIDB/04436/2020 and UIDP/04436/2020, and by project NORTE-08-5369-FSE-000039 co-founded by the European Social Fund FSE and through National funds NORTE 2020 and Regional Operacional Programa of North 2014/2020. The University of Vigo work was funded by a Xunta de Galicia grant to the Biological Oceanography Research Group (Consolidación e estruturación de unidades). This output reflects only the views of the authors, and the program authorities cannot be held responsible for any use that may be made of the information contained therei

The network BiodiversityKnowledge in practice: insights from three trial assessments

In order to develop BiodiversityKnowledge, a Network of Knowledge working at the European science–policy interface for biodiversity and ecosystem services, we conducted three trial assessments. Their purpose was to test structure and processes of the knowledge synthesis function and to produce knowledge syntheses. The trial assessments covered conservation and management of kelp ecosystems, biological control of agricultural pests, and conservation and multifunctional management of floodplains. Following the BiodiversityKnowledge processes, we set up expert consultations, systematic reviews, and collaborative adaptive management procedures in collaboration with requesters, policy and decision-makers, stakeholders, and knowledge holders. Outputs included expert consultations, systematic review protocols, a group model and a policy brief. Important lessons learned were firstly that the scoping process, in which requesters and experts iteratively negotiate the scope, scale and synthesis methodology, is of paramount importance to maximize the scientific credibility and policy relevance of the output. Secondly, selection of a broad array of experts with diverse and complementary skills (including multidisciplinary background and a broad geographical coverage) and participation of all relevant stakeholders is crucial to ensure an adequate breath of expertise, better methodological choices, and maximal uptake of outcomes: Thirdly, as the most important challenge was expert and stakeholder engagement, a high visibility and reputation of BiodiversityKnowledge, supported by an incentive system for participation, will be crucial to ensure such engagement. We conclude that BiodiversityKnowledge has potential for a good performance in delivering assessments, but it requires adequate funding, trust-building among knowledge holders and stakeholders, and a proactive and robust interface with the policy and decision making communityPeer reviewe

No evidence that wild red deer (Cervus elaphus) on the Iberian Peninsula are a reservoir of Mycobacterium avium subspecies paratuberculosis infection

The potential role of red deer (Cervus elaphus) as a reservoir of Mycobacterium avium subspecies paratuberculosis (MAP) infection is largely unknown. A total of 332 wild red deer were investigated using post-mortem examination, bacteriology and serology. Only three animals (1.12%) were found to have lesions on histopathological examination and no MAP bacteria were recovered on culture. The results suggest it is unlikely that wild red deer make a significant contribution to the maintenance of MAP infection in the region. The cross-reactivity of the ELISAs used indicates this diagnostic modality is ineffective in the detection of MAP infection in this species. The implications of these results for the control of this important pathogen in both livestock and wildlife are discussed

La Cuevona de Avín (Avín, Asturias, North Spain): A new Late Pleistocene site in the lower valley of the River Güeña

The archaeological investigations carried out in the last twenty years in the Lower Valley of the River Güeña (Asturias, central part of northern Spain) have documented different prehistoric sites, particularly with Middle and Upper Palaeolithic occupations. This paper presents the first results of the archaeological excavation carried out in the cave of La Cuevona de Avín. From the systematic study of the biotic and abiotic remains, a total of three occupation phases (Phases 1 to 3) have been determined, dated in the Late Pleistocene. The lithic studies indicate the use of local raw materials (mainly quartzite), but also regional ones (different types of flint) in the whole sequence. Retouched implements are typologically representative only during the Upper Magdalenian (Phase II) and use-wear analysis indicates the manufacture and use of artefacts in situ during this phase. Archaeozoological studies reveal continuity in subsistence strategies throughout the sequence, noting specialization in red deer hunting during the Azilian (Phase I), and more diversified prey in the older phases of the sequence. © 2022 The Author(s