Band 3 is an anchor protein and a target for SHP-2 tyrosine phosphatase in human erythrocytes

AbstractTyr phosphorylation of the multifunctional transmembrane protein band 3 has been implicated in several erythrocyte functions and disorders. We previously demonstrated that pervanadate treatment of human erythrocytes induces band-3 Tyr phosphorylation, which is catalyzed by the sequential action of tyrosine kinase Syk and tyrosine kinase(s) belonging to the Src family. In this study, we show that Tyr phosphorylation of band 3, elicited by pervanadate, N-ethylmaleimide, or diamide, greatly increases band-3 interaction with the tyrosine phosphatase SHP-2 in parallel with the translocation of SHP-2 to erythrocyte membranes. These events seem to be mediated by Src-like catalyzed phosphorylation of band 3 because both SHP-2 translocation to cellular membranes and its interaction with Tyr-phosphorylated protein are greatly counteracted by PP2, a specific inhibitor of Src kinases. Binding-competition experiments demonstrate that SHP-2 recruitment to band 3 occurs via its SH2 domain(s). In particular, our data support the view that SHP-2 docks specifically with P-Y359 of band 3. Experiments performed with intact erythrocytes in the presence of the SHP-2 inhibitor calpeptin suggest that, once recruited to Tyr-phosphorylated band 3, the tyrosine phosphatase dephosphorylates the protein. P-Y8, 21, and 904 are the residues affected by SHP-2, as judged by 32P-peptide mapping of band 3 digested with trypsin. These results indicate that in treated erythrocytes, recruitment of cytosolic SHP-2 to band 3 is a prerequisite for the subsequent dephosphorylation of the transmembrane protein

Dal segno al suono. Indagine sulla lettura musicale allo strumento

"From sign to sound" is a research project aimed to investigate some peculiar features of the musical reading with reference to instrumental performance. Its purpose is to extend the present knowledge about cognitive, musical, and pedagogical aspects of the subject, pointing out problematic issues and solutions significant for the young musicians' music performance education. The main study profile, which differentiates it from very few existing international studies on the subject, arises from the particular situation of the Latin countries (to which some other Latin American countries must be added) that adopted, starting from XIX century, the syllabic designation of notes, abandoning the alphabetic one. The aim of the research was to clarify a) the extent to which the outlined situation is widespread in Italian and European instrumental teaching, b) what are the prevailing opinions about it, and c) whether different perspectives can be found based on instrumental families or educational schools.Published"Dal segno al suono" è un progetto di ricerca che indaga alcuni aspetti peculiari della lettura musicale riferiti specificamente all'esecuzione strumentale. Il suo scopo è ampliare le attuali conoscenze sugli aspetti cognitivi, musicali e pedagogici in materia, evidenziando problematiche e soluzioni significative per l'educazione all'esecuzione musicale dei giovani musicisti. Il profilo peculiare di questo studio, che lo differenzia da pochissimi studi internazionali esistenti sull'argomento, nasce dalla particolare situazione dei paesi latini (a cui vanno aggiunti alcuni paesi latinoamericani) che adottarono, a partire dal XIX secolo, la nominazione sillabica delle note, abbandonando quella alfabetica. Lo scopo della ricerca è stato quello di chiarire a) in che misura questa situazione si trova nella didattica strumentale italiana ed europea, b) quali sono le opinioni prevalenti in merito, e c) se si possono rintracciare prospettive diverse sulla base di famiglie strumentali o scuole pedagogiche differenti

Astaxanthin Prevents Human Papillomavirus L1 Protein Binding in Human Sperm Membranes

Astaxanthin (Asta), red pigment of the carotenoid family, is known for its anti-oxidant, anti-cancer, anti-diabetic, and anti-inflammatory properties. In this study, we evaluated the effects of Asta on isolated human sperm in the presence of human papillomavirus (HPV) 16 capsid protein, L1. Sperm, purified by gradient separation, were treated with HPV16-L1 in both a dose and time-dependent manner in the absence or presence of 30 min-Asta pre-incubation. Effects of HPV16-L1 alone after Asta pre-incubation were evaluated by rafts (CTB) and Lyn dislocation, Tyr-phosphorylation (Tyr-P) of the head, percentages of acrosome-reacted cells (ARC) and endogenous reactive oxygen species (ROS) generation. Sperm membranes were also analyzed for the HPV16-L1 content. Results show that HPV16-L1 drastically reduced membrane rearrangement with percentage of sperm showing head CTB and Lyn displacement decreasing from 72% to 15.8%, and from 63.1% to 13.9%, respectively. Accordingly, both Tyr-P of the head and ARC decreased from 68.4% to 10.2%, and from 65.7% to 14.6%, respectively. Asta pre-incubation prevented this drop and restored values of the percentage of ARC up to 40.8%. No alteration was found in either the ROS generation curve or sperm motility. In conclusion, Asta is able to preserve sperm by reducing the amount of HPV16-L1 bound onto membranes

TRATAMENTO DE EFLUENTES DE HOSPITAL VETERINÁRIO POR MEIO DO USO DE BIODISCOS

O Hospital Veterinário da Pontifícia Universidade Católica do Paraná, campus Toledo, apresenta um sistema de esgoto contendo fossas sépticas para os efluentes oriundos de banheiros, cozinha, lavagem de salas, centro cirúrgico, lavagem do piso onde ficam alguns animais e onde acontecem aulas práticas. Todas as substancias químicas são descartadas em recipientes específicos para cada tipo. Com tudo, o objetivo do estudo é avaliar o desempenho de um protótipo de biodiscos aplicado no tratamento dos efluentes. Diante disso o trabalho foi realizado no laboratório de Análises Ambientais da Pontifícia Universidade Católica do Paraná Campus Toledo-PR onde foi desenvolvido um sistema de tratamento de efluente com reator de biodiscos, em escala laboratorial. A unidade piloto consiste em um recipiente contendo um conjunto de discos de policloreto de vinila (PVC), dispostas em paralelo, formando um cilindro, que, suportadas por um eixo central, são ligadas a um motor de 12 W que promove a rotação do conjunto. O mesmo ficou em operação constante durante 8 dias, e não apresentou falhas construtivas. Os materiais escolhidos para os discos foram manta abrasiva e PVC, os quais se mostraram eficientes, entretanto a manta abrasiva destacou-se na avaliação da maioria dos parâmetros como N, P e DQO. No tratamento com PVC, apresentou boa remoção em alguns parâmetros, apesar de não visualizar presença de microrganismos e bactérias (biofilme) aderidas no mesmo. &nbsp

Interactional positioning and narrative self-construction in the first session of psychodynamic-interpersonal psychotherapy

The purpose of this study is to identify possible session one indicators of end of treatment psychotherapy outcome using the framework of three types of interactional positioning; client’s self-positioning, client’s positioning between narrated self and different partners, and the positioning between client and therapist. Three successful cases of 8-session psychodynamic-interpersonal (PI) therapy were selected on the basis of client Beck Depression Inventory scores. One unsuccessful case was also selected against which identified patterns could be tested. The successful clients were more descriptive about their problems and demonstrated active rapport-building, while the therapist used positionings expressed by the client in order to explore the positionings developed between them during therapy. The unsuccessful case was characterized by lack of positive self-comment, minimization of agentic self-capacity, and empathy-disrupting narrative confusions. We conclude that the theory of interactional positioning has been useful in identifying patterns worth exploring as early indicators of success in PI therapy

Persistent and transient olfactory deficits in COVID-19 are associated to inflammation and zinc homeostasis

IntroductionThe Coronavirus Disease 2019 (COVID-19) is mainly a respiratory syndrome that can affect multiple organ systems, causing a variety of symptoms. Among the most common and characteristic symptoms are deficits in smell and taste perception, which may last for weeks/months after COVID-19 diagnosis owing to mechanisms that are not fully elucidated. MethodsIn order to identify the determinants of olfactory symptom persistence, we obtained olfactory mucosa (OM) from 21 subjects, grouped according to clinical criteria: i) with persistent olfactory symptoms; ii) with transient olfactory symptoms; iii) without olfactory symptoms; and iv) non-COVID-19 controls. Cells from the olfactory mucosa were harvested for transcriptome analyses. Results and discussionRNA-Seq assays showed that gene expression levels are altered for a long time after infection. The expression profile of micro RNAs appeared significantly altered after infection, but no relationship with olfactory symptoms was found. On the other hand, patients with persistent olfactory deficits displayed increased levels of expression of genes involved in the inflammatory response and zinc homeostasis, suggesting an association with persistent or transient olfactory deficits in individuals who experienced SARS-CoV-2 infection

Targeted activation of the SHP-1/PP2A signaling axis elicits apoptosis of chronic lymphocytic leukemia cells

Lyn, a member of the Src family of kinases, is a key factor in the dys-regulation of survival and apoptotic pathways of malignant B cells in chronic lymphocytic leukemia. One of the effects of Lyn's action is spatial and functional segregation of the tyrosine phosphatase SHP-1 into two pools, one beneath the plasma membrane in an active state promoting pro-survival signals, the other in the cytosol in an inhibited conformation and unable to counter the elevated level of cytosolic tyrosine phosphorylation. We herein show that SHP-1 activity can be elicited directly by nintedanib, an agent also known as a triple angiokinase inhibitor, circumventing the phospho-S591-dependent inhibition of the phosphatase, leading to the dephosphorylation of pro-apoptotic players such as procaspase-8 and serine/threonine phosphatase 2A, eventually triggering apoptosis. Furthermore, the activation of PP2A by using MP07-66, a novel FTY720 analog, stimulated SHP-1 activity via dephosphorylation of phospho-S591, which unveiled the existence of a positive feedback signaling loop involving the two phosphatases. In addition to providing further insights into the molecular basis of this disease, our findings indicate that the PP2A/SHP-1 axis may emerge as an attractive, novel target for the development of alternative strategies in the treatment of chronic lymphocytic leukemia

Comparative analysis of the intestinal microbiota in patients with exocrine pancreatic insufficiency of various severity

Background. Exocrine pancreatic insufficiency (EPI) is a critical host factor in determining the composition of the gut microbiota. Diseases that cause exocrine insufficiency can affect the gut microbiome, which can potentiate disease progression and complications. To date, the relationship of exocrine insufficiency in various pancreatic (PA) pathologies, in chronic pancreatitis (CP), with dysbiotic changes in the intestinal microbiota (IM) has not been reliably studied. The available data are heterogeneous and contradictory, which determines the need for further research. Aim. To conduct a comparative analysis of the taxonomic composition of the intestinal microbiota in patients with CP of various etiologies, without or with the presence of EPI of varying severity, as well as patients with severe EPI with a history of surgical intervention (SI) on the pancreas. Materials and methods. A total of 85 patients were included in the study. Patients were divided into groups according to the severity of EPI: Group 1 (n=16) patients with CP without EPI; Group 2 (n=11) patients with CP and mild EPI; Group 3 (n=17) patients with severe CP and EPI; Group 4 (n=41) severe EPI in persons with a history of SI on the pancreas. Verification of CP was carried out according to clinical, anamnestic and instrumental data. The degree of EPI was determined by the level of pancreatic elastase-1 (PE-1) feces. Informed consent for the study was obtained for each patient, an anamnesis was collected, physical and laboratory examinations were performed, and a stool sample was obtained. DNA was extracted from each stool sample, the taxonomic composition of BM was determined by sequencing the bacterial 16S rRNA genes, followed by bioinformatic analysis. Results. We followed the changes in the gut microbiota from a group of patients with CP without EPI to a group with severe EPI, in those who underwent SI. At the level of the phylum, the IM of all groups showed the dominance of Firmicutes, with the lowest representation in the severe EPI group, both with SI and CP, and the growth of the Actinobacteria, Verrucomicrobiota and Fusobacteria types. The differential representation of childbirth varied: in patients with severe EPI and CP, compared with mild, statistically significant genera Akkermansia, Ruminococcus gauvreauii group and Holdemanella; compared with CP without exocrine insufficiency, Prevotella, Ruminococcus gauvreauii group, Peptostreptococcus and Blautia dominated. The CP group with mild EPI was dominated by the following genera: Lachnospiraceae_ND 2004 group, Faecalitalea, Fusobacterium, Catenibacterium, Roseburia, Atopobium, Cloacibacillus, Clostridium innococum group, Ruminococcus torques group. All groups showed a low diversity of taxa with a predominance of opportunistic flora, including participants in oncogenesis. Conclusion. The results of the study show that patients with CP of various etiologies and patients with severe EPI who underwent specific intervention on the pancreas have intestinal microbiota dysbiosis, the severity of which is significantly influenced by the degree of EPI

Lyn sustains oncogenic signaling in chronic lymphocytic leukemia by strengthening SET-mediated inhibition of PP2A.

Aberrant protein kinase activities, and the consequent dramatic increase of Ser/Thr and -Tyr phosphorylation, promote the deregulation of the survival pathways in chronic lymphocytic leukemia (CLL), which is crucial to the pathogenesis and progression of the disease. In this study, we show that the tumor suppressor Protein Phosphatase 2A (PP2A), one of the major Ser/Thr phosphatase, is in an inhibited form due to the synergistic contribution of two events, the interaction with its physiological inhibitor SET and the phosphorylation of Y307 of the catalytic subunit of PP2A. The latter event is mediated by Lyn, a Src family kinase previously found to be overexpressed, delocalized and constitutively active in CLL cells. This Lyn/PP2A axis accounts for the persistent high level of phosphorylation of the phosphatase's targets and represents a key connection linking phosphotyrosine- and phosphoserine/threonine-mediated oncogenic signals. The data herein presented show that the disruption of the SET/PP2A complex by a novel FTY720-analogue (MP07-66) devoid of immunosuppressive effects leads to the reactivation of PP2A, which in turn triggers apoptosis of CLL cells. When used in combination with SFK inhibitors, the action of MP07-66 is synergistically amplified, providing a new option in the therapeutic strategy for CLL patients

Helicobacter pylori Diagnostic Tests Used in Europe : Results of over 34,000 Patients from the European Registry on Helicobacter pylori Management

Funding Information: This study was funded by Richen; however, clinical data were not accessible and the company was not involved in any stage of the Hp-EuReg study (design, data collection, statistical analysis, or manuscript writing). We want to thank Richen for their support. This project was promoted and funded by the European Helicobacter and Microbiota Study Group (EHMSG), the Spanish Association of Gastroenterology (AEG) and the Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd). The Hp-EuReg was co-funded by the European Union programme HORIZON (grant agreement number 101095359) and supported by the UK Research and Innovation (grant agreement number 10058099). The Hp-EuReg was co-funded by the European Union programme EU4Health (grant agreement number 101101252). Acknowledgments We want to especially thank Sylva-Astrik Torossian for her assistance in language editing. Natalia García Morales is the first author who is acting as the submission’s guarantor. All authors approved the final version of the manuscript.Peer reviewedPublisher PD