Development of a production process for a recombinant protein pneumococcal vaccine

Pneumonia represents important infection in young children (under five years old). Even with efficient vaccines, this disease was responsible for 992,000 deaths in 2015 and it was globally equivalent to 15% of all deaths in children. Bio-Manguinhos/Oswaldo Cruz Foundation (Brazil), supplies in their National Immunization Program the 10-vallent pneumococcal vaccine. This vaccine contains 10 of the 92 serotypes of this pathogen, and it´s obtained through fermentations and purifications of the capsular polysaccharides of each serotype, followed by chemical couplings to specific carrier proteins. This work presents a proposal for industrial process of only one recombinant protein antigen, the PsaA (pneumococcal surface adhesion A with 37 kDa), potentially capable to act against most of all prevalent serotypes of streptococcus pneumoniae. Based on protein expression in prokaryotic system and purification, this process reached high purity levels and yield superior to conventional fermentation systems. Biomass was obtained in bench scale reactor and studies of expression times verified that it could be reduced in 60% from presented previously. Moreover, in purification steps was observed good resolution using anion exchange (DEAE Sepharose FF®). For desalting, it was observed that gel filtration technique could be replaced for tangential cross flow filtration, better to scale-up, and using 10 kDa membranes. The developed process resulted in overall yield of 35-40%, based in BCA analysis. Electrophoretic analysis (SDS-PAGE) of the purified sample showed a band with molecular weight of 37 kDa corresponding to 80% of identified proteins. As formulation proposal, chitosan in different concentrations were used to prepare capsules of the antigen resulting in encapsulation efficiency between 26 and 36% that can be evaluated as an intranasal delivery system. This data show that the methodology developed is promising for use in industrial process aimed at obtain a recombinant vaccine

Los moluscos del cesped algal intermareale en Azores

The molluscan fauna of the high intertidal algal turf on the island of Pico (Azores) was studied in order to complement existing descriptions of the intertidal biota on these islands. A total of 15,275 specimens belonging to 19 species were found. Five species, the gastropods Alvania mediolittoralis Gofas, 1989, Omalogyra atomus (Philippi, 1841), Pisinna glabrata (Megerle von Muhlfeld, 1824) and Skeneopsis planorbis (Fabricius 0., 1780), and the bivalve Lasaea adansoni (Gmelin, 1791), account for 98% of the total number of specimens, P. glabrata itself being responsible for 46% of all specimens. In places where patellid limpets are absent, Ihe algal turf extends higher in the intertidal zone. In such places, three very abundant molluscan species characterize molluscan assemblages in the algal turf: Lasaea adansoni, Skeneopsis planorbis and Pisinna glabrata, Lasaeo adansoni and Skeneopsis planorbis virtually disappear in the transition from algal lurf to algal fronds, the only (very) abundant species being P. glabrata and, with less importance, Omalogyra atomus and Sinezona cingulata (Costa O.G., 1861)

In a community-based setting spondyloarthritis patients report higher levels of physical disability than chronic low back pain patients-results from EpiReuma.pt

Publisher Copyright: © 2023, Sociedade Portuguesa de Reumatologia. All rights reserved.Objectives: Chronic low back pain (CLBP) is a common health problem and in most patients it is not possible to identify a specific cause (non-specific CLBP). Spondyloarthritis is a musculoskeletal disorder characterized by (often inflammatory) back pain and spinal stiffness. The impact of CLBP and spondyloarthritis on patients’ physical function may be different. This study aims to compare physical disability in patients with spondyloarthritis and CLBP, in a population-based setting. Furthermore, we aim to identify modifiable risk factors for physical disability among these two populations. Methods: Data from EpiReumaPt, a national health cohort with 10 661 individuals, conducted from September 2011 to December 2013, was used. Physical function was accessed by the Health Assessment Questionnaire Disability Index (HAQ-DI) and by the physical function dimension of the 36-Item Short Form Survey (SF-36). Univariable and multivariable linear regression analyses were used to assess the differences between groups. Factors associated with physical disability were explored for both diseases. Results: We evaluated 92 patients with spondyloarthritis, 1376 patients with CLBP and 679 subjects without rheumatic and musculoskeletal diseases (RMDs). Spondyloarthritis and CLBP patients reported significantly higher levels of disability in HAQ-DI (b=0.33; p<0.001 and b=0.20; p<0.001, respectively) than subjects without RMDs. In comparison to CLBP patients, spondyloarthritis patients reported higher disability (b=0.14; p=0.03). The physical domains of SF-36, bodily pain and general health, where more affected in spondyloarthritis patients than in CLBP patients (b=-6.61; p=0.02 and b=-5.94; p=0.001, respectively). Spondyloarthritis and CLBP patients had a worse physical summary score (PCS) than mental summary score (MCS), and only PCS was significantly worse in comparison to subjects without RMDs. Factors associated with physical disability in CLBP were low back pain intensity, older age, obesity, multimorbidity, and retirement. Similarly, in spondyloarthritis physical disability was associated with retirement and multimorbidity. Factors associated with lower disability were alcohol consumption and male gender in CLBP, and regular physical exercise was associated with lower disability in both disorders. Conclusions: In this nationwide cohort, spondyloarthritis and CLBP patients reported significant physical disability. Regular physical exercise was associated with lower disability in both diseases.publishersversionpublishe

Progression of Ganglion Cell-Inner Plexiform layer thickness in the initial stages of diabetic retinopathy in type 2 diabetic patients: a 5-year longitudinal study

Diabetic Retinopathy (DR)is a frequent complication of DiabetesMellitus (DM) andthe main cause of vision loss in the working population in western countries. Diabetic Retinopathy has always been considered a microvascular disease, but it has been suggested that neurodegeneration is also associated with this complex pathology[1], although there is evidence indicating that the neurodegenerative process may progress independently[2]. To evaluate this potential association, we have examined the progression of neurodegeneration over a 5-year period of follow-up (considering thinning of ganglion cell + inner plexiform retinal layers (GCL+IPL) in individuals with type 2 diabetes (T2D) and nonproliferative DR) and explored whetheritis associated with microaneurysmturnover (MAT), diseaselevel at baseline and severity progression.info:eu-repo/semantics/publishedVersio

A transcriptomic approach to the metabolism of tetrapyrrolic photosensitizers in a marine annelid

Funding Information: Funding: The authors also acknowledge DPGM (the Portuguese General Directorate for Marine Policy) for funding the MARVEN project (ref. FA_05_2017_007). The Portuguese Foundation for Science and Technology (FCT) funded project WormALL (PTDC/BTA-BTA/28650/2017), which includes the contract attributed to M.D. and the fellowship attributed to A.P.R. UCIBIO and LAQV are financed by national funds from FCT, UID/Multi/04378/2020 and UID/QUI/50006/2020, respectively. Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland.The past decade has seen growing interest in marine natural pigments for biotechnological applications. One of the most abundant classes of biological pigments is the tetrapyrroles, which are prized targets due their photodynamic properties; porphyrins are the best known examples of this group. Many animal porphyrinoids and other tetrapyrroles are produced through heme metabolic pathways, the best known of which are the bile pigments biliverdin and bilirubin. Eulalia is a marine Polychaeta characterized by its bright green coloration resulting from a remarkably wide range of greenish and yellowish tetrapyrroles, some of which have promising photodynamic properties. The present study combined metabolomics based on HPLC-DAD with RNA-seq transcriptomics to investigate the molecular pathways of porphyrinoid metabolism by comparing the worm’s proboscis and epidermis, which display distinct pigmentation patterns. The results showed that pigments are endogenous and seemingly heme-derived. The worm possesses homologs in both organs for genes encoding enzymes involved in heme metabolism such as ALAD, FECH, UROS, and PPOX. However, the findings also indicate that variants of the canonical enzymes of the heme biosynthesis pathway can be species-and organ-specific. These differences between molecular networks contribute to explain not only the differential pigmentation patterns between organs, but also the worm’s variety of novel endogenous tetrapyrrolic compounds.publishersversionpublishe

Geographic variation in phenology behavior and response to drought of cork oak populations is crucial to cope with climate change

Cork oak is widely distributed in the Western Mediterranean region, spanning a range of different environmental conditions, and frequently dominating open woodlands of high conservation and socioeconomic value. Cork oak woodlands spread-out for over 715000 ha in Portugal, covering 21% of the national forested area and 30% of the world’s cork producing area. Cork oak is well adapted to the seasonal drought of Mediterranean climate, following several decades of warming- up and frequent drought years. However, since the 70's, maximum and minimum temperatures have risen in Portugal about 0.5 ºC each decade, corresponding to twice the average world temperature increment. In addition, since longer, more frequent, and more intense drought periods are expected, stress caused by the expansion of arid and semi-arid climate throughout the country will affect the species distribution. Consequently, not only established stands may be prone to tree mortality, but also the current reforestation effort may be jeopardized by low survival rates attributed to the use of unsuitable genetic material. It is expected that, through genetic adaptation and/or phenotypic plasticity, cork oak populations may have developed significant differences in fitness and the traits related to it. In this context, provenance trials are the best resource of material to assess the variability between and within populations from seed sources sampled in a wide range of locations (stands) covering the geographical distribution of the species. This will enable to assess the levels and patterns of genetic variation for growth and traits determining adaptation to a specific environment, hence providing crucial information to select appropriate seed sources for planting, as well as to develop sustainable breeding and gene conservation programs. In 1998, multi-site provenance trials were established at three locations in Portugal, as a part of the “European Network for the Evaluation of Genetic Resources of Cork Oak for Appropriate Use in Breeding and Gene Conservation Strategies”. Results from these field trials at 7 years from planting pointed out that seed origin must be considered in reforestation programs, since survival, growth and adaptive traits (phenology, water use efficiency - wue) showed significant differences among populations. Responses to drought (a major limitation to cork oak regeneration) of four contrasting populations, selected according to their field performance for growth, phenology and wue, were further studied under controlled-environment conditions where drought stress was induced. In addition, morphological (total height, root/shoot biomass), physiological (wue, water potential, relative water content and gas exchange) and biochemical (maximum quantum yield of PSII) traits were assessed in 6-month seedlings produced from acorns collected in the stands representing the four contrasting provenances. Results from the controlled experiment provided an indication that drought adaptation was mainly related to early stomatal closure and root investment, with these traits showing significant differences between the studied populations. We have also observed significant differences in growth rhythm, as well as dissimilar temporal drought responses, in the populations under study. In conclusion, both field trials and controlled environment showed consistent results and indicated an important influence of geographic origin on growth performance and wue.projeto PTDC/AGR-AAM/104364/200

Combining central pattern generators with the electromagnetism-like algorithm for head motion stabilization during quadruped robot locomotion

Visually-guided locomotion is important for autonomous robotics. However, there are several difficulties, for instance, the head shaking that results from the robot locomotion itself that constraints stable image acquisition and the possibility to rely on that information to act accordingly. In this article, we propose a controller architecture that is able to generate locomotion for a quadruped robot and to generate head motion able to minimize the head motion induced by locomotion itself. The movement controllers are biologically inspired in the concept of Central Pattern Generators (CPGs). CPGs are modelled based on nonlinear dynamical systems, coupled Hopf oscillators. This approach allows to explicitly specify parameters such as amplitude, offset and frequency of movement and to smoothly modulate the generated oscillations according to changes in these parameters. We take advantage of this particularity and propose a combined approach to generate head movement stabilization on a quadruped robot, using CPGs and a global optimization algorithm. The best set of parameters that generates the head movement are computed by the electromagnetism-like algorithm in order to reduce the head shaking caused by locomotion. Experimental results on a simulated AIBO robot demonstrate that the proposed approach generates head movement that does not eliminate but reduces the one induced by locomotion

Retinal neurodegeneration in different risk phenotypes of diabetic retinal disease

Diabetic retinopathy (DR) has been considered a microvascular disease, but it has become evident that neurodegeneration also plays a key role in this complex pathology. Indeed, this complexity is reflected in its progression which occurs at different rates in different type 2 diabetic (T2D) individuals. Based on this concept, our group has identified three DR progression phenotypes that might reflect the interindividual differences: phenotype A, characterized by low microaneurysm turnover (MAT <6), phenotype B, low MAT (<6) and increased central retinal thickness (CRT); and phenotype C, with high MAT (≥6). In this study, we evaluated the progression of DR neurodegeneration, considering ganglion cell+inner plexiform layers (GCL+IPL) thinning, in 170 T2D individuals followed for a period of 5 years, to explore associations with disease progression or risk phenotypes. Ophthalmological examinations were performed at baseline, first 6 months, and annually. GCL+IPL average thickness was evaluated by optical coherence tomography (OCT). Microaneurysm turnover (MAT) was evaluated using the RetMarkerDR. ETDRS level and severity progression were assessed in seven-field color fundus photography. In the overall population there was a significant loss in GCL+IPL (−0.147 μm/year), independently of glycated hemoglobin, age, sex, and duration of diabetes. Interestingly, this progressive thinning in GCL + IPL reached higher values in phenotypes B and C (−0.249 and −0.238 μm/year, respectively), whereas phenotype A remained relatively stable. The presence of neurodegeneration in all phenotypes suggests that it is the retinal vascular response to the early neurodegenerative changes that determines the course of the retinopathy in each individual. Therefore, classification of different DR phenotypes appears to offer relevant clarification of DR disease progression and an opportunity for improved management of each T2D individual with DR, thus playing a valuable role for the implementation of personalized medicine in DR.info:eu-repo/semantics/publishedVersio

Photoinactivation of bacterial and fungal planktonic/biofilm forms using the combination of a porphyrinic formulation with potassium iodide

Antimicrobial photodynamic therapy (aPDT) is a promising approach against multidrug-resistant microorganisms. In this work, we accessed the photodynamic efficiency of an affordable formulation composed of five cationic porphyrins (FORM) and its combined effect with potassium iodide (KI) on a large spectrum of microorganisms. For this purpose, the aPDT assays were conducted with FORM alone and FORM + KI on planktonic and biofilm forms of Gram(+) (Staphylococcus aureus) and Gram(−) (Escherichia coli) bacteria and of the yeast Candida albicans. The results obtained indicate that FORM, at low concentrations (0.5–5.0 μM), had an efficient photodynamic action on the planktonic forms of E. coli, S. aureus, and C. albicans. Moreover, the combination of FORM with KI improved the photodynamic action of this PS, promoting microbial inactivation with lower PS concentrations and treatment time. The combination of FORM + KI was also extremely efficient in the destruction of bacterial and fungal biofilms. This outstanding effect may be due to the action of longer-lived iodine reactive species produced by the reaction of KI with the ROS generated by FORM during the aPDT treatment.info:eu-repo/semantics/publishedVersio

Different risk profiles for progression of nonproliferative diabetic retinopathy: a 2-year study

Characterization of 2-year progression of different risk phenotypes in eyes with mild and moderate nonproliferative diabetic retinopathy (NPDR) in type 2 diabetes (T2D). A 2-year prospective longitudinal cohort study (CORDIS, NCT03696810) was conducted. Ophthalmological examinations were performed including best corrected visual acuity, color fundus photography and optical coherence tomography (OCT and OCTA). OCT metrics, central retinal thickness and ganglion cell layer + inner plexiform layer (GCL + IPL) thickness were analyzed. OCTA metrics, vessel density (VD), perfusion density (PD) and area of intercapillary spaces (AIS) were obtained from superficial and deep capillary plexus (SCP, DCP). Only phenotype C identified by decreased VD ≥ 2 SD of healthy controls and phenotype B identified by subclinical macular edema with decreased VD < 2 SD of healthy controls were included. One hundred twenty-two eyes from T2D individuals were included in study; 65 eyes (53%) were classified as phenotype B and 57 eyes (47%) as phenotype C. For phenotype B, progression was associated with thinning of the GCL + IPL (ETDRS 35, 1 year p = 0.013, 2 year p < 0.001; ETDRS 43–47, 2 year p = 0.003) and vessel closure involving mainly the DCP for both ETDRS grades (ETDRS 35, 1 year p = 0.025, 2 year p = 0.034; ETDRS 43–47, 1 year p = 0.011). For phenotype C there was also progressive thinning of the GCL + IPL (ETDRS 35, in both years p ≤ 0.001; ETDRS 43–47, 1 year p = 0.002, 2 year p = 0.001), with vessel closure involving mainly SCP (ETDRS 35, 1 year p = 0.012, 2 year p = 0.023 in full-retina), which appeared to stabilize at maximal values in ETDRS grade 43–47 at the end of 2 years. ETDRS severity changes at the end of the 2-year period showed that worsening was associated with phenotype C with changes involving predominantly the SCP (VD, p = 0.005; PD, p = 0.008; AIS, p = 0.005). Association between ETDRS classification of NPDR severity and identification of different risk phenotypes offers new perspective to predict disease progression in T2D individuals with NPDR.info:eu-repo/semantics/publishedVersio