Biochemische Analyse der Interaktion von P2X4 und P2X7 und Charakterisierung von P2X7B und genetisch verlinkten P2X4/P2X7-Konkatameren

Das übergeordnete Ziel meiner Arbeit war die Untersuchung der Interaktion von P2X7 und P2X4. Zur Identifikation von potenziellen Interaktionspartnern habe ich den P2X7-Rezeptors von Mensch und Ratten mit den weiteren P2X-Isoformen koexprimiert und nach dem Köder-und Beute-System proteinchemisch analysiert. Eine Heterotrimerisierung von hP2X7 und hP2X4 konnte ich durch Koexpression der C-terminal trunkierten hP2X7-Untereinheit (hP2X71-408 bzw. hP2X71-436) mit der hP2X4-Untereinheit nachweisen. Die Ergebnisse deuten somit daraufhin, dass die hP2X7A-Untereinheit ebenso mit der hP2X4-Untereinheit zu einem Heterotrimer assemblieren würde, da die entscheidende Assemblierungsdomäne in der Ektodomäne liegt. Zur Erstellung der homomeren und heteromeren Konkatamere wurden die rP2X4- und rP2X7-Untereinheiten in definierter Zusammensetzung aneinander ligiert. Es wurde biochemisch mittels SDS-PAGE untersucht, welchen Einfluss die Linker, die Anordnung der Untereinheiten und die Protease-Inhibitoren auf die Expression der Konkatamere haben. Die Untersuchung zeigte, dass lange Linker für eine starke Expression und kurze Linker für eine verringerte Erzeugung von Abbau-Produkten am besten geeignet sind. Ein letztes Ziel der Arbeit war die Charakterisierung der desensibilisierenden Spleißvariante hP2X7B. Die unterschiedliche Kinetik und der Einfluss der Länge der C-terminalen Endodomäne auf die Kinetik-Eigenschaften der beiden hP2X7-Varianten wurde dargestellt. Um einen direkten Vergleich zu erzielen wurde die C-terminale Endodomäne der hP2X7A-Untereinheit um die abweichende Aminosäuresequenz gekürzt und die unterschiedliche Expression der beiden hP2X7-Varianten biochemisch mittels SDS-PAGE und BN-PAGE sowie elektrophysiologisch mittels der Zwei-Elektroden-Spannungsklemme charakterisiert. Die Desensibilisierung der hP2X7B-Untereinheit bei der ersten ATP-Applikation spricht für die Bildung der zytoplasmatischen Kappe, trotz einer C-terminalen Endodomäne bestehend aus neun Aminosäuren. Zusammengenommen wurde die Heteromerisierung der beiden Untereinheiten P2X4 und P2X7 biochemisch nachgewiesen und homomere P2X4- und P2X7-Konkatamere sowie P2X7B biochemisch verifiziert

Hubungan Pemberian Asi Eksklusif Dengan Tumbuh Kembang Bayi Usia 6-12 Bulan Di Wilayah Kerja Puskesmas Kaubun Tahun 2023

Bayi mengalami proses tumbuh kembang yang dipengaruhi oleh beberapa faktor, salah satunya adalah gizi. Unsur gizi pada bayi dapat dipenuhi dengan pemberian ASI, bahkan sampai umur 6 bulan sesuai rekomendasi WHO diberikan ASI eksklusif. Berdasarkan data pemberian ASI Eksklusif di Puskesmas Kaubun tahun 2020 sebesar 42,35%, kurang dari target awal sebesar 45%. Artinya target belum tercapai. Tujuan penelitian ini yaitu mengetahui hubungan pemberian ASI Eksklusif dengan tumbuh kembang bayi usia 6 bulan di Wilayah Kerja Puskesmas Kaubun Tahun 2023. Desain penelitian yang digunakan peneliti dalam penelitian ini adalah studi korelasi (correlation study) dengan menggunakan pendekatan cross sectional. Populasi dalam penelitian ini adalah ibu yang memiliki balita usia6-12 bulan yang berkunjung di Puskesmas Kaubun sejumlah 36 ibu, tekhnik sampling yang digunakan total sampling. Instrument dalam penelitian ini yaitu timbangan berat badan, alat ukur tinggi badan, lembar ceklis, KSPS. Analisa data menggunakan uji Chi-Square. Hasil penelitian menunjukkan bahwa nilai signifikasi p-value = 0,001 dengan nilai odd ratio = 12,467 (Pertumbuhan) dan p-value = 0,000 (Perkembangan). Adanya hubungan pemberian ASI Eksklusif dengan tumbuh kembang bayi usia 6 bulan di Wilayah Kerja Puskesmas Kaubun Tahun 202

The Evaluation of English Coursebook for Young Learners When English Rings a Bell: A Survey Research

This research aimed to assess When English Rings a Bell, an introductory English textbook. In addition, this research seeks to examine one textbook used in seventh-grade classrooms. This research aims to determine if the goals of the English textbook When English Rings a Bell align with the principles of teaching English to young students. When English Rings a Bell, the researcher, the evaluation checklist from (Bilash, 2009), and the textbook's material served as the research instruments. Overall, the textbook seems like it would be a good fit for young learners in terms of several aspects, such as General Appearance, Layout and Design, Activities, and Language skills.). In addition, the textbook fulfills the principles of teaching for young learners. Keywords: Evaluation, English Coursebook, Young Learners

Preparation and Evaluation of Controlled Release Calcium Alginate Beads Containing Mefenamic Acid

The present study was designed to investigate the effects of different variables on the release profile of mefenamic acid from calcium alginate beads formulated using modified emulsification method. Five formulations of beads (F1-F5) with different drug: polymer ratios were prepared in which the amount of drug (1g) was kept constant and the amount of calcium alginate was increased from 1 up to 5g. All the formulated beads were evaluated for percentage yield, entrapped efficiency and in vitro release profile at pH 1.2 and 7.2 dissolution media. For further minimize the release of drug from calcium alginate beads, the selected formulation F3 beads were coated separately with 3% and 6% Eudragit S100 solution using dipping method. The coated beads (F6 and F7) minimized the release of drug both in pH 1.2 and 7.2. The study confirms significant effects of two variables, drug:polymer ratio and coating on the release of drug from beads which can be effectively utilized to control the release from prepared calcium alginate formulations

ADAPTACIÓN A LOS CAMBIOS PSICOSOCIALES GENERADOS POR LA ERRADICACIÓN DE CULTIVOS ILÍCITOS EN LOS HABITANTES DE LA VEREDA LA PANGUANA II, RETORNO – GUAVIARE

El presente estudio tiene como objetivo, analizar el proceso de adaptación a los cambios psicosociales generados por la erradicación de cultivos ilícitos en los habitantes de la vereda la Panguana II, Retorno-Guaviare, por medio de la aplicación de técnicas cualitativas, que permitan la creación de un plan de intervención que contribuya al desarrollo funcional de la población. En la recolección de los datos se tuvieron en cuenta ocho personas pertenecientes al Programa Nacional Integral de Sustitución de Cultivos Ilícitos (PNIS). Asimismo, se utilizaron las técnicas de entrevista, grupo focal e historia de vida. El análisis de estos se realizó mediante la triangulación de técnicas con una descripción profunda de cada categoría (adaptación, cambios psicosociales y acuerdo de paz). Basándonos en lo planteado anteriormente, se halló como resultado que la vereda La Panguana II Retorno – Guaviare está conformada por 22 familias, y que una de las modificaciones primordiales es la estructura económica, sin dejar a un lado los cambios familiares, sociales y personales. Por otra parte, hacen énfasis en el incumplimiento del Estado en relación al programa PNIS del cual son participes por la influencia familiar y social. Finalmente, en relación a la adaptación, se haya como resultado que la mayoría de los participantes no han vivenciado este proceso por diversos factores. También en los resultados se establece un plan de intervención para fortalecer la adaptación a los cambios psicosociales que han experimentado

ETHYLENE RESPONSE FACTOR 115 integrates jasmonate and cytokinin signaling machineries to repress adventitious rooting in Arabidopsis

Adventitious root initiation (ARI) is ade novoorganogenesis program and a key adaptive trait in plants. Several hormones regulate ARI but the underlying genetic architecture that integrates the hormonal crosstalk governing this process remains largely elusive. In this study, we use genetics, genome editing, transcriptomics, hormone profiling and cell biological approaches to demonstrate a crucial role played by the APETALA2/ETHYLENE RESPONSE FACTOR 115 transcription factor. We demonstrate that ERF115 functions as a repressor of ARI by activating the cytokinin (CK) signaling machinery. We also demonstrate thatERF115is transcriptionally activated by jasmonate (JA), an oxylipin-derived phytohormone, which represses ARI in NINJA-dependent and independent manners. Our data indicate that NINJA-dependent JA signaling in pericycle cells blocks early events of ARI. Altogether, our results reveal a previously unreported molecular network involving cooperative crosstalk between JA and CK machineries that represses ARI

Propuesta estratégica de mejora en la implementación de los estándares mínimos del sistema de gestión de la seguridad y salud en el trabajo (sg-sst) en la empresa gsi group para el año 2020.

Desarrollo normativo del SGSST, Resultados de evaluacion estandandares minimos SG-SST GSI GROUP, Plan de mejora a la implementación del SG-SST, Diagrama de gantt, Tabla de asignación de responsabilidadesPropuesta estratégica de mejora en la implementación de los estándares mínimos del sistema de gestión de la seguridad y salud en el trabajo (sg-sst) en la empresa gsi group para el año 2020, y así lograr una correcta y eficaz aplicación de las normas y leyes que regulan la seguridad y salud en el trabajo en la empresa GSI GROUP.Strategic proposal for improvement in the implementation of the minimum standards of the Occupational Health and Safety Management System (SGS-SST) in the company Gsi Group for the year 2020, and thus achieve a correct and effective application of the standards and laws that regulate occupational safety and health in the company GSI GROUP

Bioengineering commensal bacteria-derived outer membrane vesicles for delivery of biologics to the gastrointestinal and respiratory tract.

Gram-negative bacteria naturally produce and secrete nanosized outer membrane vesicles (OMVs). In the human gastrointestinal tract, OMVs produced by commensal Gram-negative bacteria can mediate interactions amongst host cells (including between epithelial cells and immune cells) and maintain microbial homeostasis. This OMV-mediated pathway for host-microbe interactions could be exploited to deliver biologically active proteins to the body. To test this we engineered the Gram-negative bacterium Bacteroides thetaiotaomicron (Bt), a prominent member of the intestinal microbiota of all animals, to incorporate bacteria-, virus- and human-derived proteins into its OMVs. We then used the engineered Bt OMVs to deliver these proteins to the respiratory and gastrointestinal (GI)-tract to protect against infection, tissue inflammation and injury. Our findings demonstrate the ability to express and package both Salmonella enterica ser. Typhimurium-derived vaccine antigens and influenza A virus (IAV)-derived vaccine antigens within or on the outer membrane of Bt OMVs. These antigens were in a form capable of eliciting antigen-specific immune and antibody responses in both mucosal tissues and systemically. Furthermore, immunisation with OMVs containing the core stalk region of the IAV H5 hemagglutinin from an H5N1 strain induced heterotypic protection in mice to a 10-fold lethal dose of an unrelated subtype (H1N1) of IAV. We also showed that OMVs could express the human therapeutic protein, keratinocyte growth factor-2 (KGF-2), in a stable form that, when delivered orally, reduced disease severity and promoted intestinal epithelial repair and recovery in animals administered colitis-inducing dextran sodium sulfate. Collectively, our data demonstrates the utility and effectiveness of using Bt OMVs as a mucosal biologics and drug delivery platform technology

Bioengineering commensal bacteria-derived outer membrane vesicles for delivery of biologics to the gastrointestinal and respiratory tract

Gram-negative bacteria naturally produce and secrete nanosized outer membrane vesicles (OMVs). In the human gastrointestinal tract, OMVs produced by commensal Gram-negative bacteria can mediate interactions amongst host cells (including between epithelial cells and immune cells) and maintain microbial homeostasis. This OMV-mediated pathway for host-microbe interactions could be exploited to deliver biologically active proteins to the body. To test this we engineered the Gram-negative bacterium Bacteroides thetaiotaomicron (Bt), a prominent member of the intestinal microbiota of all animals, to incorporate bacteria-, virus- and human-derived proteins into its OMVs. We then used the engineered Bt OMVs to deliver these proteins to the respiratory and gastrointestinal (GI)-tract to protect against infection, tissue inflammation and injury. Our findings demonstrate the ability to express and package both Salmonella enterica ser. Typhimurium-derived vaccine antigens and influenza A virus (IAV)-derived vaccine antigens within or on the outer membrane of Bt OMVs. These antigens were in a form capable of eliciting antigen-specific immune and antibody responses in both mucosal tissues and systemically. Furthermore, immunisation with OMVs containing the core stalk region of the IAV H5 hemagglutinin from an H5N1 strain induced heterotypic protection in mice to a 10-fold lethal dose of an unrelated subtype (H1N1) of IAV. We also showed that OMVs could express the human therapeutic protein, keratinocyte growth factor-2 (KGF-2), in a stable form that, when delivered orally, reduced disease severity and promoted intestinal epithelial repair and recovery in animals administered colitis-inducing dextran sodium sulfate. Collectively, our data demonstrates the utility and effectiveness of using Bt OMVs as a mucosal biologics and drug delivery platform technology