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Group A Streptococcus (GAS) infections are responsible for significant morbidity and mortality worldwide. The outlook for an effective global vaccine is reduced because of significant antigenic variation among GAS strains worldwide. Other challenges in GAS therapy include the lack of common access to antibiotics in developing countries, as well as allergy to and treatment failures with penicillin and increasing erythromycin resistance in the industrialized world. At the portal of entry, GAS binds to newly deposited extracellular matrix, which is rich in cellular fibronectin isoforms with extra domain A (EDA, also termed EIIIA) via the surface adhesin, the streptococcal collagen-like protein 1 (Scl1). Recombinant Scl1 constructs, derived from diverse GAS strains, bind the EDA loop segment situated between the C and C’ β-strands. Despite the sequence diversity in Scl1 proteins, multiple sequence alignments and secondary structure predictions of Scl1 variants, as well as crystallography and homology modeling studies, point to a conserved mechanism of Scl1-EDA binding. We propose that targeting this interaction may prevent the progression of infection. A synthetic cyclic peptide, derived from the EDA C-C’ loop, binds to recombinant Scl1 with a micromolar dissociation constant. This review highlights the current concept of EDA binding to Scl1 and provides incentives to exploit this binding to treat GAS infections and wound colonization.

OBJECTIVES: Changes in body weight, length and head circumference have long been indicators of infant growth and development. PediaTrac™ is a web-based measure designed to engage caregivers in the gathering of longitudinal, real-time, multi-domain data on infant/toddler growth and development at time periods corresponding to well-child visits. The purpose of this study was to assess the changes in growth (weight, length, head circumference) between the newborn and 6 m time periods for both term and pre-term (

Prokaryotic proteins with extended collagen domain are found in many bacterial species that are pathogenic to humans and animals. The collagen domain is often fused to additional ligand-binding domains and plays both structural and functional roles in modular "bacterial collagens." Here, we describe the step-by-step expression and purification of the recombinant streptococcal collagen-like proteins, rScl, using the Strep-tag II system. The integrity and structural characterization of recombinant collagen-like proteins is very important for defining their function.

Bacterial efflux pumps are an important pathogenicity trait because they extrude a variety of xenobiotics. Our laboratory previously identified in silico Burkholderia collagen-like protein 8 (Bucl8) in the hazardous pathogens Burkholderia pseudomallei and Burkholderia mallei. We hypothesize that Bucl8, which contains two predicted tandem outer membrane efflux pump domains, is a component of a putative efflux pump. Unique to Bucl8, as compared to other outer membrane proteins, is the presence of an extended extracellular region containing a collagen-like (CL) domain and a non-collagenous C-terminus (Ct). Molecular modeling and circular dichroism spectroscopy with a recombinant protein, corresponding to this extracellular CL-Ct portion of Bucl8, demonstrated that it adopts a collagen triple helix, whereas functional assays screening for Bucl8 ligands identified binding to fibrinogen. Bioinformatic analysis of the bucl8 gene locus revealed it resembles a classical efflux-pump operon. The bucl8 gene is co-localized with downstream fusCDE genes encoding fusaric acid (FA) resistance, and with an upstream gene, designated as fusR, encoding a LysR-type transcriptional regulator. Using reverse transcriptase (RT)-qPCR, we defined the boundaries and transcriptional organization of the fusR-bucl8-fusCDE operon. We found exogenous FA induced bucl8 transcription over 80-fold in B. pseudomallei, while deletion of the entire bucl8 locus decreased the minimum inhibitory concentration of FA 4-fold in its isogenic mutant. We furthermore showed that the putative Bucl8-associated pump expressed in the heterologous Escherichia coli host confers FA resistance. On the contrary, the Bucl8-associated pump did not confer resistance to a panel of clinically-relevant antimicrobials in Burkholderia and E. coli. We finally demonstrated that deletion of the bucl8-locus drastically affects the growth of the mutant in L-broth. We determined that Bucl8 is a component of a novel tetrapartite efflux pump, which confers FA resistance, fibrinogen binding, and optimal growth.

In this work an alternative TOFD method is presented, capable to detect and size defects in the inspected material with good precision. It was shown that the diffraction used in this technique is the most relevant signal among the longitudinal backwall and the shear backwall echoes, compared to other TOFD methods. The proposed technique showed to be particularly efficient in cases where the coventional TOFD method does not perform well; cases include near-surface defects, cracks under compressive stress and bottom tip of defects. The test configuration and a mathematical description referred to the wave path of the signal of interest is described and confirmed by numerical simulations and dynamic and static tests. The proposed method was tested for different defect depth location and find industrial applications such as inspection of cracks in tubes, closed cracks and weld joints. It opens a new possibility for TOFD based inspections.

Keratinized epidermis constitutes a powerful barrier of the mucosa and skin, effectively preventing bacterial invasion, unless it is wounded and no longer protective. Wound healing involves deposition of distinct extracellular matrix (ECM) proteins enriched in cellular fibronectin (cFn) isoforms containing extra domain A (EDA). The streptococcal collagen-like protein 1 (Scl1) is a surface adhesin of group A Streptococcus (GAS), which contains an N-terminal variable (V) domain and a C-terminally located collagen-like domain. During wound infection, Scl1 selectively binds EDA/cFn isoforms and laminin, as well as low-density lipoprotein (LDL), through its V domain. The trimeric V domain has a six-helical bundle fold composed of three pairs of anti-parallel α-helices interconnected by hypervariable loops, but the roles of these structures in EDA/cFn binding are unclear. Here, using recombinant Scl (rScl) constructs to investigate structure–function determinants of the Scl1–EDA/cFn interaction, we found that full-length rScl1, containing both the globular V and the collagen domains, is necessary for EDA/cFn binding. We established that the surface-exposed loops, interconnecting conserved α-helices, guide recognition and binding of Scl1-V to EDA and binding to laminin and LDL. Moreover, electrostatic surface potential models of the Scl1-V domains pointed to a conserved, negatively charged pocket, surrounded by positively charged and neutral regions, as a determining factor for the binding. In light of these findings, we propose an updated model of EDA/cFn recognition by the Scl1 adhesin from GAS, representing a significant step in understanding the Scl1–ECM interactions within the wound microenvironment that underlie GAS pathogenesis.

We as ECO-sofía Thinking School see this Conference as the opportunity to present our research advances in the proposal to build a free society and bioethics. The ECO-Sofia School tries to theoretically configure a bioethical society as an alternative life for the global society in the face of models and proposals that are built on the basis of reductionist visions that come from mechanistic sources whose interpretation and understanding of life are circumscribes to give biased and incomplete views of the person and society.From the name of ECO-sofia we want to express that the value, meaning and horizon of life are determined by the ethical, social and universal knowledge that constitute, in our understanding, the pillars of the future bioethical society for which the problem of life focuses and focuses on "thinking and understanding life from life itself". From our theoretical horizon the world is integral, universal and obeys the cosmovisions that derive from the complex thinking, since that world is not something homogeneous but a complex and multidimensional heterogeneity that must be interpreted from the open rationality to reach a deep understanding of its dynamics and structure, that allows to generate the changes and transformations required in order to build a bioethical society that harmonizes the values and principles that determine it with the concern for biodiversity and life in all its manifestations Nosotros como Escuela de Pensamiento ECO-sofía vemos esta Conferencia como la oportunidad de presentar nuestros avances de investigación en la propuesta de construcción de una sociedad libre y bioética. La Escuela ECO-sofía trata de configurar, teóricamente, una sociedad bioética como una alternativa de vida para la sociedad global frente a modelos y propuestas que se erigen sobre la base de visiones reduccionistas que provienen de fuentes mecanicistas cuya interpretación y comprensión de la vida se circunscribe a dar miradas sesgadas e incompletas de la persona y sociedad. A partir de la denominación ECO-sofia queremos expresar que el valor, el sentido y el horizonte de la vida están determinados por los saberes éticos, sociales y universales que constituyen, en nuestra comprensión, los pilares de la futura sociedad bioética para la cual el problema de la vida se centra y se enfoca en “pensar y comprender la vida desde la vida misma”. Desde nuestro horizonte teórico el mundo es integral, universal y obedece a las cosmovisiones que derivan del pensamiento complejo, en cuanto ese mundo no es algo homogéneo sino una heterogeneidad compleja y multidimensional quedebe ser interpretado desde la racionalidad abierta para alcanzar una comprensión profunda de su dinámica y estructura, que permita generar los cambios y transformaciones requeridos a fin de construir una sociedad bioética que armonice los valores y principios que la determinan con la preocupación por la biodiversidad y la vida en todas sus manifestaciones

The collagen domain, which is defined by the presence of the Gly-X-Y triplet repeats, is amongst the most versatile and widespread known structures found in proteins from organisms representing all three domains of life. The streptococcal collagen-like (Scl) proteins are widely present in pathogenic streptococci, including Streptococcus pyogenes, S. agalactiae, S. pneumoniae, and S. equi. Experiments and bioinformatic analyses support the hypothesis that all Scl proteins are homotrimeric and cell wall-anchored. These proteins contain the rod-shaped collagenous domain proximal to cell surface, as well as a variety of outermost non-collagenous domains that generally lack predicted functions but can be grouped into one of six clusters based on sequence similarity. The well-characterized Scl1 proteins of S. pyogenes show a dichotomous switch in ligand binding between human tissue and blood environments. In tissue, Scl1 adhesin specifically recognizes the wound microenvironment, promotes adhesion and biofilm formation, decreases bacterial killing by neutrophil extracellular traps, and modulates S. pyogenes virulence. In blood, ligands include components of complement and coagulation-fibrinolytic systems, as well as plasma lipoproteins. In all, the Scl proteins signify a large family of structurally related surface proteins, which contribute to the ability of streptococci to colonize and cause diseases in humans and animals.

This article is a proposal to grasp territory from a different perspective, in coexistence with the traditional and planned city. With, however, the physical discoveries of the 20th century – particularly those related to quantum physics and the theory of relativity– it became evident that space-time are inseparable magnitudes, in contraction with the tenetof Euclidean geometry and opening at the same time the possibility of discussing territory in terms of new utopias about reality. Drawing upon such utopias about reality, this means that there are no inherent hierarchical structures ruled by a fixed center; neither is there an absolute reference framework as expressed by Newtonian theory. From this point of view, the research projects a territorial transformation strategy that gives rise to new forms of self-organization in order to understand territory as a fabric and not as mere coexistence, resulting in the need to look for a new complex and polycentric space., Este artículo es una propuesta para mirar al territorio desde otra perspectiva, en coexistencia con la ciudad tradicional y planificada. Sin embargo, con los descubrimientos físicos del siglo XX, especialmente los relacionados con la teoría cuántica y de la relatividad, es evidente que el espacio-tiempo son magnitudes inseparables, lo que contraviene y postula la geometría euclidiana a la vez que abre posibilidad para hablar sobre el territorio en términos de nuevas utopías sobre la realidad. Ello implica que, desde esas utopías de la realidad, no existe una estructura jerárquica inherente que se rija por un centro fijo, y tampoco se reconoce marco de referencia absoluto expresado por la teoría newtoniana. Desde esta perspectiva, la investigación proyecta una estrategia de transformación territorial que da origen a nuevas formas de auto-organización, con el fin de entender el territorio como un tejido y no concebirlo como mera coexistencia, y, por tanto, se debe buscar un nuevo espacio policéntrico complejo.

За підсумком кількох сезонів досліджень спільної українсько-польської експедиції за участю кафедри архітектури та реставрації Національного університету “Львівська політехнікаˮ виявлено руїни літописної катедральної церкви Пречистої Богородиці ХІІІ ст. у Холмі, спорудженої за короля Данила Романовича. У статті повідомлено про застосування спеціальної методики досліджень, проаналізовано окремі архітектурно-археологічні та історичні й архітектурні аспекти нововиявленої пам’ятки, звернено увагу на її синтетичні риси, у яких поєднувалися риси візантійської (хрестово-банева структура) та західноєвропейської романської архітектури: окремі планувальні особливості (прямокутне завершення вівтарної частини) та модерні технологічні прийоми (цегляне мурування у вендійській системі). Автори здійснили спробу гіпотетичної графічної реконструції об’єкта на перший період його функціонування. During 2013–2017 years the Polish-Ukrainian science project has been carried out with the aim of recognize the cathedral church of the Virgin Mary (Bogorodytsia) built in Kholm by Danylo Romanovych in the middle of the XIII-th cent. Interdisciplinary, non-invasion prospections as well as archaeological excavations were done with a view to finding and identifying the location of the cathedral church, known until present only from the written sources. Analysis results of the archaeological researches allow us to propose a hypothesis on the original architectonic and planning structure of this cathedral. It was 14 m wide and near 22.4 m long. It was east-west oriented. The distance between the outer walls and the pillars was 1.75–2.0 m, the inside diameter of the dome on the pillars was about 5.0 m. The architectural-spatial layout of the church was based of two lengthened rectangular parts –three-aisled four-pillared part and the apse. The foundation walls were made of sandstone bonded with chalky-clayey and sand-lime mortars and was about 2.0 m wide. The width of the wall above the corbel was 1.22–1.24 m. The wall thickness, the way of placing bricks, firm and of high quality mortar indicate that the central part of the structure could have been high. The walls of the church were made of the brick (260–270 × 120–125 × 80–85 mm) in the Romanesque masonry technique. The outer wall surface was not plastered and the outer joints between bricks were well-made. The remains of the walls show that they were built without pilasters. The main entrance to the temple was in the middle part of the western wall in the form of a perspective portal decorated with framing. In the lower part from the outside the portal was made of bricks and from the inside part it was framed with the white lime blocks. The preserved fragments of a block of stone indicate that it had a carved profile. The technology of the church wall building differs significantly from the technology used at the same time in other towns of Rus. It is one of the first bigger cathedral structures of Eastern (Ortodox) Rite built in new technology. According to archeological and architectural data as well as comparing its results to the analysis of the written sources, we may single out two main phases of the history of the Orthodox church built by Daniel Romanovich. Phase No.1 – building the ortodox church after the town fire and the beginning of its existence (from 1256 up to the 15th century). The sanctuary was then continuously used for sacred purposes and for burying the dead. An example of great respect for the chronologically oldest graves is the maintenance of burial No. 26 located in the south aisle, which was covered with an arch connecting the pillar and the wall while the temple was later converted (the construction was made of Gothic bond but from Roman brick). Phase No. 2 – from the 16th to the beginning of the 17th century. In the times of bishop Filaret (1507– 1533) the ortodox church was significantly rebuilt. The west wall of the church and two pillars were pulled down. A new wall in the Gothic tradition with the entrance portal was built at that place. In 1573 a new wooden narthex with a belfry was erected. The results of the research show that the first cathedral in Kholm, its architectural and building techniques represents the innovative technology for its time – the middle of the XIII-th cent. on the Eastern Europe.

Video game studies pertaining to representations of gender, race, culture, and sexuality are commonly discussed in relation to the characters, narrative, or gameplay mechanics. Video game environments are often left out of these discussions as most publications focus on how the environment was created and environmental storytelling. In this pilot study the researcher had participants analyze and discuss three futuristic, dystopian environments that contained alternative ways of representing gender, race, culture, and sexuality. The environments did not have gameplay or character interactions so that the player could focus on the environment. Results indicated that futuristic dystopian environments do not need to rely on stereotypes and harmful depictions to evoke core themes, even if they relate to violence, sex, oppression, or addiction. However, a lack of representation or a “neutral” take on representating gender, race, sexuality, and culture results in world that feels bland, isolated, and unnatural. Additionally, while participants disagreed on certain representations or noticed different aspects of the environments, the researcher found that focusing on “playtesting” just the environment can reveal how certain values, themes, and representations arise out of prop repetition, prop juxtaposition, and even how the player moves physically through the space.

Summary The human‐adapted pathogen group A Streptococcus (GAS) utilizes wounds as portals of entry into host tissue, wherein surface adhesins interact with the extracellular matrix, enabling bacterial colonization. The streptococcal collagen‐like protein 1 (Scl1) is a major adhesin of GAS that selectively binds to two fibronectin type III (FnIII) repeats within cellular fibronectin, specifically the alternatively spliced extra domains A and B, and the FnIII repeats within tenascin‐C. Binding to FnIII repeats was mediated through conserved structural determinants present within the Scl1 globular domain and facilitated GAS adherence and biofilm formation. Isoforms of cellular fibronectin that contain extra domains A and B, as well as tenascin‐C, are present for several days in the wound extracellular matrix. Scl1‐FnIII binding is therefore an example of GAS adaptation to the host's wound environment. Similarly, cellular fibronectin isoforms and tenascin‐C are present in the tumor microenvironment. Consistent with this, FnIII repeats mediate GAS attachment to and enhancement of biofilm formation on matrices deposited by cancer‐associated fibroblasts and osteosarcoma cells. These data collectively support the premise for utilization of the Scl1‐FnIII interaction as a novel method of anti‐neoplastic targeting in the tumor microenvironment.

OBJECTIVES: Achieving sensorimotor (SEM) milestones is one measure of early infant brain development promoted through higher docosahexaenoic acid (DHA) levels. Both breastmilk and formula contain DHA, with a global level of 0.32% in breastmilk and many formula brands. However, a 2017 study of Midwestern U.S. mothers found significantly lower breastmilk DHA levels. It was thus hypothesized that infants fed formula would have higher SEM scores. The objective of this study was to measure differences in SEM development between infants fed breastmilk, formula, or a combination in participants of the PediaTrac™ Project. PediaTrac is a web-based measure providing longitudinal, real time, multidomain data on infant and toddler growth and development at time periods corresponding to well child visits. METHODS: Using PediaTrac, data were collected from 548 caregiver-infant dyads across multiple Midwestern sites. Caregivers reported the primary nutrition source as breastfeeding, formula or combination. Their responses to gross and fine motor function questions were used to create a SEM composite, Percent of Maximum Possible (POMP) score at newborn (NB), 2-, 4- and 6-months. Data were analyzed via ANOVA and Tukey test using SPSS. RESULTS: Infants fed formula had statistically higher mean SEM scores at NB (M = 0.618(Formula), M = 0.590(Breast), p = 0.017), 2- (M = 0.706(Formula), M = 0.680(Breast), p = 0.006) and 6-months (M = 0.727(Formula), M = 0.696(Breast), p = 0.014) compared to breastfed infants. Mean SEM scores of combination fed infants were higher than breastfed infants at 2- (M = 0.701(Combination), M = 0.680(Breast), p = 0.184), 4- (M = 0.684(Combination,) M = 0.673(Breast,) p = 0.573) and 6-months (M = 0.704(Combination), M = 0.696(Breast), p = 0.895), despite no significant differences between scores. CONCLUSIONS: Formula fed infants showed consistently higher SEM scores than breastfed infants in the first 6-months. These findings contradict previous research, indicating a need for further investigation into variables contributing to these discrepancies such as maternal breastmilk DHA concentrations, socioeconomic factors, site specific confounds, accuracy of parent reports of motor development, etc. FUNDING SOURCES: Eunice Kennedy Shriver National Institute of Child Health & Human Development of the National Institutes of Health. EMU College of Health & Human Services Research Support Award.

This paper introduces a method for an ultrasonic imaging with a phased array based on a wave migration algorithm. The method allows for imaging layered objects with lateral velocity variations such as objects with a complex geometry or layers that are not perpendicular to the array's axis. The full-matrix capture ensures that there is enough information to reconstruct an image even when the wave indication angle is large. The method is implemented in a omega-k domain. The proposed algorithm is first tested in a single simulation of a concave object with side drilled holes under the concave surface. For evaluating the algorithm's performance three experiments are presented: one with a tilted object (surface not perpendicular with respect to the array axis) with side drilled holes and two experiments of an object with concave surface and two artificial defects under it. The results presented in the paper verify that the proposed method reconstructs images from the data gathered with the phased array.

This is an advance of the Project entitled: Comprehensions and Practices of theIntegral Formation. My intention is to present the truth as the value of great importance in the key of the complex thought of St Tomas de Aquino. With this offer one tries to present a Reading of St Tomas who re-lives the paper of thinking, through a unifying way, aboutthe ethical, philosophical problems, educational projects and the contributions from the religious thought, to construct the outstanding individual copromised with the problems of this world and opened for the transcendence which isnot left out of the real. Esta ponencia presenta es avance del Proyecto titulado Comprensiones Y Prácticas De La Formación Integral. Mi intención es presentar la verdad como el valor de gran importancia enla clave del pensamiento complejo de Santo Tomás de Aquino. Con esta propuesta se pretende presentar una lectura de Santo Tomas que reviva el papel de pensar de manera unificadora los problemas éticos, filosóficos, proyectos educativos, y aportes desdeel pensamiento religioso para construir la persona íntegra, comprometida con los problemas de este mundo y abierta a la transcendencia la cual no es apartada de lo real.

The arsenal of virulence factors deployed by streptococci includes streptococcal collagen-like (Scl) proteins. These proteins, which are characterized by a globular domain and a collagen-like domain, play key roles in host adhesion, host immune defense evasion, and biofilm formation. In this work, we demonstrate that the Scl2.3 protein is expressed on the surface of invasive M3-type strain MGAS315 of Streptococcus pyogenes. We report the crystal structure of Scl2.3 globular domain, the first of any Scl. This structure shows a novel fold among collagen trimerization domains of either bacterial or human origin. Despite there being low sequence identity, we observed that Scl2.3 globular domain structurally resembles the gp41 subunit of the envelope glycoprotein from human immunodeficiency virus type 1, an essential subunit for viral fusion to human T cells. We combined crystallographic data with modeling and molecular dynamics techniques to gather information on the entire lollipop-like Scl2.3 structure. Molecular dynamics data evidence a high flexibility of Scl2.3 with remarkable interdomain motions that are likely instrumental to the protein biological function in mediating adhesive or immune-modulatory functions in host-pathogen interactions. Altogether, our results provide molecular tools for the understanding of Scl-mediated streptococcal pathogenesis and important structural insights for the future design of small molecular inhibitors of streptococcal invasion.