X-ray structure of the mature ectodomain of phogrin

Phogrin/IA-2β and ICA512/IA-2 are two paralogs receptor-type protein-tyrosine phosphatases (RPTP) that localize in secretory granules of various neuroendocrine cells. In pancreatic islet β-cells, they participate in the regulation of insulin secretion, ensuring proper granulogenesis, and β-cell proliferation. The role of their cytoplasmic tail has been partially unveiled, while that of their luminal region remains unclear. To advance the understanding of its structure-function relationship, the X-ray structure of the mature ectodomain of phogrin (ME phogrin) at pH 7.4 and 4.6 has been solved at 1.95- and 2.01-Å resolution, respectively. Similarly to the ME of ICA512, ME phogrin adopts a ferredoxin-like fold: a sheet of four antiparallel β-strands packed against two α-helices. Sequence conservation among vertebrates, plants and insects suggests that the structural similarity extends to all the receptor family. Crystallized ME phogrin is monomeric, in agreement with solution studies but in striking contrast with the behavior of homodimeric ME ICA512. The structural details that may cause the quaternary structure differences are analyzed. The results provide a basis for building models of the overall orientation and oligomerization state of the receptor in biological membranes

Educator Perceptions Of Social-Emotional Learning Embedded In Academic Instruction

This qualitative exploratory case study examined the results of the CASEL (2021) School-based Staff Survey on SEL Implementation and individual semi-structured interviews with six teachers and three staff members regarding social-emotional learning (SEL) at the secondary school level. The purpose of this qualitative exploratory case study was to explore how secondary teachers and staff at a mid-size public, suburban high school in Connecticut perceive the implementation of the RULER SEL (Brackett et al., 2019) program. There were four themes that emerged from the data: becoming a warm demander (Hammond, 2015), adapting RULER SEL tools, RULER SEL and future impact, and the need for increased individual support. The first theme included the subtheme of building dynamic relationships with students. The second theme included the subtheme of professional development. Based on the findings three recommendations were made. First, schools should provide professional development opportunities for teachers and staff focused on adopting a warm demander (Hammond, 2015) approach in the classroom. Second, longitudinal studies were recommended to evaluate the long-term impact of RULER SEL (Brackett et al., 2019) on student outcomes, well-being, and academic achievement. Third, collaborating with other schools, SEL experts, and community organizations was recommended to allow educators to share resources, and successful practices related to SEL implementation. Recommendation for future studies include a longitudinal study and comparative analysis of RULER SEL (Brackett et al., 2019). Keywords: Social-emotional learning, RULER SEL, warm demander, professional developmen

Numerical Model to Predict Hemolysis and Transport in a Membrane-Based Microfluidic Device

Microfluidics has become an increasingly popular tool in the design and development of medical devices and artificial organs. Two promising applications of microfluidics are dialyzers and oxygenators. As a step toward portable dialysis treatment, continuous microfluidic dialysis may resolve many clinical issues with current dialysis treatments. Additionally, commercially available oxygenators exceed the blood volume of neonatal patients; low-volume microfluidic devices may safely deliver oxygen to these patients. Two critical parameters in the development of these devices is mechanical hemolysis and membrane diffusion, which are intricately connected to the geometry, flow rate, properties of the membrane, and each other. A computational model is developed to elucidate the connection between these phenomena to guide the design and optimization of these devices. In vitro experiments are conducted to validate the model. Importantly, a subset of hemolysis models agrees with experimental data, which is consistent with the literature. Additionally, the effect of microfluidic mixing elements that perturb flow near the membrane interface are studied in silico and in vitro. These data reveal that herringbone mixing elements increase hemolysis by 10% and flux across the membrane interface by 38% in silico and a statistically significant difference between smooth and herringbone devices is observed for a subset of devices tested. Furthermore, 10 of 18 computational models of hemolysis are shown to be statistically similar to experimental data. The agreement of these results suggest that finite element analysis may be able to quantitively model important factors in the design of microfluidic oxygenators and dialyzers

Novel prokaryotic expression of thioredoxin-fused insulinoma associated protein tyrosine phosphatase 2 (IA-2), its characterization and immunodiagnostic application

Background The insulinoma associated protein tyrosine phosphatase 2 (IA-2) is one of the immunodominant autoantigens involved in the autoimmune attack to the beta-cell in Type 1 Diabetes Mellitus. In this work we have developed a complete and original process for the production and recovery of the properly folded intracellular domain of IA-2 fused to thioredoxin (TrxIA-2ic) in Escherichia coli GI698 and GI724 strains. We have also carried out the biochemical and immunochemical characterization of TrxIA-2icand design variants of non-radiometric immunoassays for the efficient detection of IA-2 autoantibodies (IA-2A). Results The main findings can be summarized in the following statements: i) TrxIA-2ic expression after 3 h of induction on GI724 strain yielded ≈ 10 mg of highly pure TrxIA-2ic/L of culture medium by a single step purification by affinity chromatography, ii) the molecular weight of TrxIA-2ic (55,358 Da) could be estimated by SDS-PAGE, size exclusion chromatography and mass spectrometry, iii) TrxIA-2ic was properly identified by western blot and mass spectrometric analysis of proteolytic digestions (63.25 % total coverage), iv) excellent immunochemical behavior of properly folded full TrxIA-2ic was legitimized by inhibition or displacement of [35S]IA-2 binding from IA-2A present in Argentinian Type 1 Diabetic patients, v) great stability over time was found under proper storage conditions and vi) low cost and environmentally harmless ELISA methods for IA-2A assessment were developed, with colorimetric or chemiluminescent detection. Conclusions E. coli GI724 strain emerged as a handy source of recombinant IA-2ic, achieving high levels of expression as a thioredoxin fusion protein, adequately validated and applicable to the development of innovative and cost-effective immunoassays for IA-2A detection in most laboratories.Fil: Guerra, Luciano Lucas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni; ArgentinaFil: Faccinetti, Natalia Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni; ArgentinaFil: Trabucchi, Aldana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni; ArgentinaFil: Rovitto, Bruno David. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni; ArgentinaFil: Sabljic, Adriana Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni; ArgentinaFil: Poskus, Edgardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni; ArgentinaFil: Iacono, Ruben Francisco. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni; ArgentinaFil: Valdez, Silvina Noemi. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni; Argentin

Retrorectal tumours: literature review and Vilnius University Hospital "Santariskiu klinikos" experience of 14 cases

<p>Abstract</p> <p>Objective</p> <p>Retrorectal tumours are rare lesions in adults. The diagnosis of retrorectal lesion is often difficult and misdiagnosis is common. We present significant number of cases in view of scarce information available on this matter.</p> <p>Methods</p> <p>14 patients were treated at Vilnius university hospital "Santariskiu klinikos" Centre of abdominal surgery from 1997 to 2010. The case notes of patients who underwent surgery for a retrorectal tumour were reviewed retrospectively. Surgical histories, operations, histological tumour type, surgical time, weight of the specimen, blood loss, length of stay were analysed.</p> <p>Results</p> <p>13 patients underwent laparotomy, 1 patient had combined perineal approach and laparotomy. The most common types of the tumour were fibroma (3 cases), leiomyosarcoma (2 cases). 5 tumours (35,7%) were found to be malignant. 57% of the patients had undergone at least one operation prior to definitive treatment. 5 female patients were initially admitted under gynaecologists. Hospital stay varied from 14 days to 22 days (mean 16,2 days). A report of a representative case is presented.</p> <p>Conclusions</p> <p>Retrorectal lesions in female patients can mimic gynaecological pathology. Patients with this rare pathology are to be treated in a major tertiary hospital by surgeons, who are able to operate safely in the retrorectal space.</p

OUT OF LEFT SCREEN: FACTORS IMPACTING WELL-BEING IN DIGITAL AND PHYSICAL COLLEGIATE ATHLETES

Competitive video gaming, or “esports,” has quickly become a popular college extracurricular. Its classification as a varsity sport on many campuses reflects a broader shift in cultural conceptualizations and perceptions of athleticism. As esports continue to evolve in today’s increasingly digital world, new professional leagues emerge, and viewership rivals that of traditional physical sports. Esports can be a gateway for unique opportunities for college students; however, it may also pose some challenges. This study explored differences between digital and physical athletes, who must balance their chosen activity with other collegiate responsibilities, including academic, social, and physical health. Physical sports and esports athletes were compared in terms of athletic identity, health behaviors, cognitive functioning, and mental health outcomes. Data from 244 athletes (179 physical sports; 65 esports) was analyzed, revealing that esports athletes reported significantly higher depressive symptoms than physical sports athletes. Esports athletes exhibited greater functional disability as well. They also outperformed physical sports athletes in cognitive tasks of visual motor speed and reaction time. Physical sports athletes, on the other hand, reported stronger athletic identity. Notably, critical predictors of depressive symptoms included athletic social role identity and sleep quality for both types of athletes; Sleep quality predicted symptoms of anxiety as well. Interestingly, higher family role rankings were linked to lower depression and anxiety symptoms. Overall, these results suggest that while esports athletes may excel in certain cognitive areas, they face greater challenges in mental health and functional disability compared to physical athletes. These findings are important because they highlight the unique mental health challenges all collegiate athletes face, emphasizing the need for tailored support systems to help them balance the demands of their chosen sport with academic, social, and physical well-being

Structural health monitoring of non-linear hysteretic structures.

Ensuring life safety is the primary design and maintenance requirement for civil structures designed to be serviceable for a specific lifetime. However, structures subjected to a number of factors may experience quicker or more gradual deterioration than anticipated, or even a premature loss of function. Scheduled visual inspection is the oldest and most commonly used damage monitoring technique, but has significant disadvantages when quick damage assessment and certainty are needed, such as after a major seismic event. Quick, accurate and quantitative determination of the damage state is vital following an earthquake, to estimate damage, remaining life-time, and ensure safe re-occupancy, if possible. Rapid development of sensor technology and increasing computing power has enabled continuous structural monitoring using various sensing techniques. The measured data can be analyzed using structural health monitoring (SHM) methods. SHM refers to all elements of the process of identifying mechanical properties of a structural system, comparing it with previous states, detecting changes/abnormalities, and relating these to damage. A successful SHM method should be able to automatically identify and locate damage after large, non-linear response events. The majority of existing, primarily vibration based, SHM techniques have serious limitations in situations where a quick, accurate, and quantitative assessment is needed. More specifically, many SHM techniques perform well when structures behave linearly and are subjected to ambient loads, but this does not apply to earthquake events. Moreover, some methods can only work off-line, involve significant computational effort and/or human input, and/or do not provide any indication of damage location and/or severity. To address these limitations, this thesis explores the application of a novel SHM implementation strategy composed of a novel modal parameter identification and its subsequent application to a proven hysteresis loop analysis (HLA) method. The study demonstrates the proposed strategy can be readily used to track the performance of non-linear degrading structures subjected to strong ground motion, essentially in real-time and without human input. Thus, the proposed tools can be used to support/replace visual inspection results, reduce downtime, minimize business disruptions and, most importantly, maximize life safety. More specifically, this thesis proposes and analyses the application of a novel modal parameter identification technique, which performs in near real-time and, most importantly, is efficient when approximating non-linear structures subjected to relatively short duration ground motion inputs. The technique operates in modal space and is based on a pre-defined optimization process, which decouples frequency response spectra of interfering, generally higher frequency, modes. Optimization can be realized over relatively short time windows to provide continuous monitoring of highly non-linear, degrading structures. In particular, identified modal parameters can be readily used to identify damage. However, modal parameters can have very poor sensitivity to damage and are often difficult to interpret. Thus, it is challenging to infer the location and severity of damage based on detected changes/variation in modal parameters alone. In this research, the identified time-varying modal parameters are used to decompose the structural response and reconstruct single mode dominant restoring force-deformation hysteresis loops, which can be readily analyzed using recently developed hysteresis loops analysis (HLA). The versatility and robustness of HLA has been explored in a number of studies. However, the analyzed case structures employed in these validation cases exhibited very small contribution from the higher modes, which typically can cause significant irregularities, and make effective implementation of HLA more problematic. Hence, this thesis aims to improve the robustness of HLA, using mode segregation and reconstruction of single mode dominant, regular shape hysteresis loops from non-linear structural response. First, this research develops a modal parameter output-only identification technique, which is validated for a simple time-invariant linear structure. Second, the output-only method is extended to an input-output method enabling operators to carry out near-real time identification of non-linear structures, which is validated for a simple time-varying non-linear structure. Third, the input-output method is validated using the simulation results of a more complex non-linear multi-degree-of-freedom structure, formulated using fiber elements. Finally, the proposed SHM strategy, consisting of continuous modal parameter identification and subsequent application of HLA is validated for two experimental non-linear structures. Overall, this thesis proposes a novel system identification technique, which performs outputonly identification of linear structures and, more importantly, provides input-output real-time modal parameter tracking of highly non-linear structures. Thus, the method extends the application of modal SHM methods to non-linear cases. The proposed technique performs successfully without operator input and can be easily automated to provide continuous modal tracking and damage detection. The technique performs both as stand-alone for damage detection and in combination with HLA for damage quantification as demonstrated for highly non-linear cases

Flow Cytometric microsphere-based immunoassay as a novel non-radiometric method for the detection of glutamic acid decarboxylase autoantibodies in type 1 Diabetes Mellitus

The first measurable sign of arising autoimmunity in Type 1 Diabetes Mellitus is the detection of autoantibodies against beta-cell antigens, such as glutamic acid decarboxylase (GAD65). GAD65 autoantibodies (GADA) are usually measured by Radioligand Binding Assay (RBA). The aim of this work was to develop protocols of Flow Cytometric microsphere-based immunoassays (FloCMIA) which involved glutamic acid decarboxylase fused to thioredoxin (TrxGAD65) adsorbed on polystyrene microspheres. Detection of bound GADA was accomplished by the use of anti-human IgG-Alexa Fluor 488 (Protocol A), anti-human IgG-biotin and streptavidindichlorotriazinyl aminofluorescein (DTAF) (Protocol B) or TrxGAD65-biotin and streptavidin- DTAF (Protocol C). Serum samples obtained from 46 patients assayed for routine autoantibodies at Servicios Tecnológicos de Alto Nivel (STAN-CONICET) were analyzed by RBA, ELISA and three alternative FloCMIA designs. Protocol C exhibited the highest specificity (97.8%) and sensitivity (97.4%) and a wide dynamic range (1.00-134.40 SDs). Samples obtained from 40 new-onset diabetic patients were also analyzed to further evaluate the performance of protocol C. The latter protocol showed a sensitivity of 58.6% and a prevalence of 47.5%. Two patients resulted positive only by FloCMIA protocol C and its SDs were higher than RBA and ELISA, showing a significantly wide dynamic range. In conclusion, FloCMIA proved to be highly sensitive and specific, requiring a low sample volume; it is environmentally adequate, innovative and it represents a cost-effective alternative to traditional GADA determination by RBA and/or ELISA; making it applicable to most medium-complexity laboratories.Fil: Guerra, Luciano Lucas. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral "Profesor R. A. Margni"; ArgentinaFil: Trabucchi, Aldana. Consejo Nacional de Investigaciones Cientiâ­ficas y Tecnicas. Oficina de Coordinacion Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral "profesor R. A. Margni"; ArgentinaFil: Faccinetti, Natalia Ines. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral "Profesor R. A. Margni"; ArgentinaFil: Iacono, Ruben Francisco. Consejo Nacional de Investigaciones Cientiâ­ficas y Tecnicas. Oficina de Coordinacion Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral "profesor R. A. Margni"; ArgentinaFil: Ureta, Daniela Beatriz. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral "Profesor R. A. Margni"; ArgentinaFil: Poskus, Edgardo. Consejo Nacional de Investigaciones Cientiâ­ficas y Tecnicas. Oficina de Coordinacion Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral "profesor R. A. Margni"; ArgentinaFil: Valdez, Silvina Noemi. Consejo Nacional de Investigaciones Cientiâ­ficas y Tecnicas. Oficina de Coordinacion Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral "profesor R. A. Margni"; Argentin

Phosphorylation of silk fibroins improves the cytocompatibility of silk fibroin derived materials: a platform for the production of tuneable material

Silk fibroin demonstrates great biocompatibility and is suitable for many biomedical applications, including tissue engineering and regenerative medicine. Current research focuses on manipulating the physico-chemical properties of fibroin, and examining the effect of this manipulation on firobin's biocompatibility. Regenerated silk fibroin was modified by in vitro enzymatic phosphorylation and cast into films. Films were produced by blending, at several ratios, the phosphorylated and un-phosphorylated fibroin solutions. Fourier transform infra-red spectroscopy was used to determine the specific P–OH vibration peak, confirming the phosphorylation of the regenerated silk fibroin solution. Differential scanning calorimetry showed that phosphorylation altered the intra- and inter-molecular interactions. Further experiments demonstrated that phosphorylation can be used to tailor the hydrophylicity/hydrophobicity ratio as well as the crystalinity of silk fibroin films. Release profiling of a model drug was highly dependent on silk modification level. Cytotoxicity assays showed that exposure to lixiviates of phosphorylated films only slightly affected cellular metabolism and proliferation, although direct contact resulted in a strong direct correlation between phosphorylation level and cell proliferation. This new method for tuning silk biomaterials to obtain specific structural and biochemical features can be adapted for a wide range of applications. Phosphorylation of silk fibroins may be applied to improve the cytocompatibility of any silk-based device that is considered to be in contact with live animals or human tissues.The authors would like to acknowledge the support granted to the authors by European NOVO Project, contract no. FP7-HEALTH 2011-two-stage 278402