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In silico Epitope Mapping of Glucose-6-Phosphate Isomerase: A Rheumatoid Arthritis Autoantigen
(2017)
Rheumatoid arthritis-like symptoms can be initiated experimentally in naive K/BxN mice by simultaneously administering the two monoclonal antibodies 11H3 and 46H9. Both antibodies specifically recognize Glucose-6-Phosphate Isomerase (GPI), a known auto antigen in RA patients. Amino acid sequences of the Fv parts of the antibodies were determined by translating the respective hybridoma DNA sequences and served for threedimensional structure modeling of the paratope regions. In silico docking of both Fv antibody structure models to the X-ray structures of the homodimeric murine GPI as well as to the homodimeric human GPI predicted the murine epitope of the 11H3 antibodies to comprise partial amino acid sequences QRVRSGDWKGYTGKS (aa134-148) and AAKDPSAVAK (aa232-241), generating an assembled (conformational) epitope. The 11H3 epitope on human GPI encompasses the matching partial amino acid sequences QRVRSGDWKGYTGKT (aa134-148) and AAKDPSAVAK (aa232-241). The epitope of the 46H9 antibody was determined to consist of the partial murine GPI amino acid sequence RKELQAAGKSPEDLEK (aa446-461) and the human GPI amino acid sequence RKELQAAGKSPEDLER (aa446-461), respectively, resembling consecutive (linear) epitopes. The predicted epitopes were verified by mass spectrometric epitope mapping using synthetic epitope peptides. Peptide QRVRSGDWKGYTGKS[GSMSGS] AAKDPSAAK included a small spacer sequence in between the epitope sequences, mimicking the assembled epitope for the 11H3 antibody. The peptide RKELQAAGKSPEDLEK represented the consecutive epitope for the 46H9 antibody. The determined B-cell epitopes of GPI and their interactions with the monoclonal antibodies provide a detailed structural understanding of immunological disease onset mechanisms in a mouse model of rheumatoid arthritis.
Ob im Studium oder in der Praxis - bei der technischen Chemie kommt man schnell an seine Grenzen. Aber keine Sorge, "Technische Chemie für Dummies" hilft Ihnen, bei diesem komplexen Thema den Durchblick zu behalten. Nach einem allgemeinen Überblick über die Entwicklungen, Herausforderungen und Konzepte der technischen Chemie und einer verständlichen Übersicht über die nötige Mathematik lernen Sie, was man bei der praktischen und theoretischen Vorarbeit beachten muss, um die chemische Reaktion später in einem größeren Maßstab durchführen zu können. Anschließend erfahren Sie alles über Reaktionsmodellierung, Katalysatoren und chemische Reaktoren. Idealisierte Modelle helfen Ihnen dabei, aber auch die Umsetzung unter realen Bedingungen kommt nicht zu kurz. Der Verfahrenstechnik ist ein eigener Teil gewidmet, damit auch Trenntechnik, Strömungsmechanik, Fluidströmungen, Dimensionierung und Co. bald kein Problem mehr für Sie sind.
Chemie ist viel einfacher, als es häufig heißt. Dieses Buch soll dazu beitragen, ihr Interesse an diesem Fach zu wecken oder zu vertiefen. Alle grundlegenden Prinzipien der Chemie werden nachvollziehbar dargestellt. Querbezüge und Zusammenhänge zwischen den verschiedenen Fachgebieten werden gezeigt. Sie werden keine Formel finden, deren Herleitung Sie nicht nachvollziehen können. Am Ende fast jeden Kapitels gibt es Übungsaufgaben. Ausführliche Lösungen gibt es natürlich auch. Das sollte nicht nur für die Prüfungen der ersten Semester reichen, sondern Ihnen auch ein sicheres Fundament für Ihr weiteres Studium bieten.
Chemie kann im Studium ganz schön kompliziert sein, besonders wenn es etwas mehr in die Tiefe geht. Dass man aber auch komplizierte Dinge leicht verständlich und bisweilen amüsant erklären kann, beweist dieses Lehrbuch. Stefanie Ortanderl und Ulf Ritgen erklären die Grundlagen der Chemie und so erfahren Sie, was Sie über Atommodelle, Bindungstypen und das Periodensystem wissen sollten.
Beta-ketothiolase deficiency, also known as mitochondrial acetoacetyl-CoA thiolase (T2) deficiency, is an autosomal recessive disease caused by mutations in the acetylCoA acetyltransferase 1 (ACAT1) gene. A German T2deficient patient that developed a severe ketoacidotic episode at the age of 11 months, was revealed to be a compound heterozygote of a previously reported null mutation, c.472A>G (p.N158D) and a novel mutation, c.949G>A (p.D317N), in ACAT1. The c.949G>A mutation was suspected to cause aberrant splicing as it is located within an exonic splicing enhancer sequence (c. 947CTGACGC) that is a potential binding site for serine/argininerich splicing factor 1. A mutation in this sequence, c.951C>T, results in exon 10 skipping. A minigene construct was synthesized that included exon 9truncated intron 9exon 10truncated intron 10exon 11, and the splicing of this minigene revealed that the c.949G>A mutant construct caused exon 10 skipping in a proportion of the transcripts. Furthermore, additional substitution of G for C at the first nucleotide of exon 10 (c.941G>C) abolished the effect of the c.949G>A mutation. Transient expression analysis of the c.949G>A mutant cDNA revealed no residual T2 activity in the mutated D317N enzyme. Therefore, c.949G>A (D317N) is a pathogenic missense mutation, and diminishes the effect of an exonic splicing enhancer and causes exon 10 skipping. The present study demonstrates that a missense mutation, or even a synonymous substitution, may disrupt enzyme function by interference with splicing.
Small Molecules Enhance Scaffold-Based Bone Grafts via Purinergic Receptor Signaling in Stem Cells
(2018)
The need for bone grafts is high, due to age-related diseases, such as tumor resections, but also accidents, risky sports, and military conflicts. The gold standard for bone grafting is the use of autografts from the iliac crest, but the limited amount of accessible material demands new sources of bone replacement. The use of mesenchymal stem cells or their descendant cells, namely osteoblast, the bone-building cells and endothelial cells for angiogenesis, combined with artificial scaffolds, is a new approach. Mesenchymal stem cells (MSCs) can be obtained from the patient themselves, or from donors, as they barely cause an immune response in the recipient. However, MSCs never fully differentiate in vitro which might lead to unwanted effects in vivo. Interestingly, purinergic receptors can positively influence the differentiation of both osteoblasts and endothelial cells, using specific artificial ligands. An overview is given on purinergic receptor signaling in the most-needed cell types involved in bone metabolism-namely osteoblasts, osteoclasts, and endothelial cells. Furthermore, different types of scaffolds and their production methods will be elucidated. Finally, recent patents on scaffold materials, as wells as purinergic receptor-influencing molecules which might impact bone grafting, are discussed.
Die Optimierung von Produktionsprozessen steht im Vordergrund jedes Produzenten, vor allem im Hinblick auf den optimalen Erntezeitpunkt. Zur Pflückreife sollen Kirschen als nicht-klimakterische Früchte eine optimale und hochwertige Fruchtqualität aufweisen, eine ausreichende Anzahl an Erntehelfern, Pflückhilfen, Transportkisten, Sortier- und Lagereinrichtungen sowie Absatzwege vorhanden sein. Aus diesem Grund entwickelten Wissenschaftler in der Vergangenheit diverse Reifeindices und Erntemodelle zur Bestimmung des optimalen Erntezeitpunkts von Früchten, erst an Äpfeln, dann für Steinobst.
Non-Destructive Sensor-Based Prediction of Maturity and Optimum Harvest Date of Sweet Cherry Fruit
(2017)
(1) Background: The aim of the study was to use innovative sensor technology for non-destructive determination and prediction of optimum harvest date (OHD), using sweet cherry as a model fruit, based on different ripening parameters. (2) Methods: Two cherry varieties in two growing systems viz. field and polytunnel in two years were employed. The fruit quality parameters such as fruit weight and size proved unsuitable to detect OHD alone due to their dependence on crop load, climatic conditions, cultural practices, and season. Coloration during cherry ripening was characterized by a complete decline of green chlorophyll and saturation of the red anthocyanins, and was measured with a portable sensor viz. spectrometer 3-4 weeks before expected harvest until 2 weeks after harvest. (3) Results: Expressed as green NDVI (normalized differential vegetation index) and red NAI (normalized anthocyanin index) values, NAI increased from -0.5 (unripe) to +0.7 to +0.8 in mature fruit and remained at this saturation level with overripe fruits, irrespective of variety, treatment, and year. A model was developed to predict the OHD, which coincided with when NDVI reached and exceeded zero and the first derivative of NAI asymptotically approached zero. (4) Conclusion: The use of this sensor technology appears suitable for several cherry varieties and growing systems to predict the optimum harvest date.
One of the primary current astrobiological goals is to understand the limits of microbial resistance to extraterrestrial conditions. Much attention is paid to ionizing radiation, since it can prevent the preservation and spread of life outside the Earth. The aim of this research was to study the impact of accelerated He ions (150 MeV/n, up to 1 kGy) as a component of the galactic cosmic rays on the black fungus C. antarcticus when mixed with Antarctic sandstones—the substratum of its natural habitat—and two Martian regolith simulants, which mimics two different evolutionary stages of Mars. The high dose of 1 kGy was used to assess the effect of dose accumulation in dormant cells within minerals, under long-term irradiation estimated on a geological time scale. The data obtained suggests that viable Earth-like microorganisms can be preserved in the dormant state in the near-surface scenario for approximately 322,000 and 110,000 Earth years within Martian regolith that mimic early and present Mars environmental conditions, respectively. In addition, the results of the study indicate the possibility of maintaining traces within regolith, as demonstrated by the identification of melanin pigments through UltraViolet-visible (UV-vis) spectrophotometric approach.
The modern concept of the evolution of Mars assumes that life could potentially have originated on the planet Mars, possibly during the end of the late heavy bombardment, and could then be transferred to other planets. Since then, physical and chemical conditions on Mars changed and now strongly limit the presence of terrestrial-like life forms. These adverse conditions include scarcity of liquid water (although brine solutions may exist), low temperature and atmospheric pressure, and cosmic radiation. Ionizing radiation is very important among these life-constraining factors because it damages DNA and other cellular components, particularly in liquid conditions where radiation-induced reactive oxidants diffuse freely. Here, we investigated the impact of high doses (up to 2 kGy) of densely-ionizing (197.6 keV/µm), space-relevant iron ions (corresponding on the irradiation that reach the uppermost layer of the Mars subsurface) on the survival of an extremophilic terrestrial organism-Cryomyces antarcticus-in liquid medium and under atmospheric conditions, through different techniques. Results showed that it survived in a metabolically active state when subjected to high doses of Fe ions and was able to repair eventual DNA damages. It implies that some terrestrial life forms can withstand prolonged exposure to space-relevant ion radiation.
People with type 2 Diabetes have an elevated risk for developing cardiovascular disease (CVD) for which dyslipidemia is the major contributor. Diabetic patients have characteristic pattern of dyslipidemia with decreased level of high density lipoprotein cholesterol (HDL-C) and elevated triglycerides (TG) level. However, in diabetes mellitus, low density lipoprotein cholesterol (LDL-C) which is used as one of the markers for the risk of CVD, is underestimated so in such cases the levels of non-High density lipoprotein cholesterol (non-HDL-C) can be a stronger predictor of CVD as it strongly correlates with atherogenic lipoproteins. Therefore, an attempt has been made to evaluate the level of non-HDL-C as a newer marker for the risk of cardiovascular disease and to fi nd out the pattern of dyslipidemia in diabetes mellitus. The present study comprised of 82 type 2 Diabetic cases and 81 non-diabetic controls. Among the diabetics, the majority of the subjects (61.0%) were HDL-C dyslipidemic. However, among the controls, the maximum numbers of individuals (40.7%) were TG dyslipidemic. Diabetics have signifi cantly elevated ratio of total cholesterol to high density lipoprotein cholesterol (TC/HDL-C) and the signifi cant increased levels of non-high density lipoprotein cholesterol (non-HDL-C) compared to controls which can be used as markers of dyslipidemia and can also be used to predict the risk of cardiovascular disease in type 2 Diabetes Mellitus.
Host-derived succinate accumulates in the airways during bacterial infection. Here, we show that luminal succinate activates murine tracheal brush (tuft) cells through a signaling cascade involving the succinate receptor 1 (SUCNR1), phospholipase Cβ2, and the cation channel transient receptor potential channel subfamily M member 5 (TRPM5). Stimulated brush cells then trigger a long-range Ca2+ wave spreading radially over the tracheal epithelium through a sequential signaling process. First, brush cells release acetylcholine, which excites nearby cells via muscarinic acetylcholine receptors. From there, the Ca2+ wave propagates through gap junction signaling, reaching also distant ciliated and secretory cells. These effector cells translate activation into enhanced ciliary activity and Cl- secretion, which are synergistic in boosting mucociliary clearance, the major innate defense mechanism of the airways. Our data establish tracheal brush cells as a central hub in triggering a global epithelial defense program in response to a danger-associated metabolite.