The Pathogenic TSH β-Subunit Variant C105Vfs114X Causes a Modified Signaling Profile at TSHR

1) Background: Central congenital hypothyroidism (CCH) is a rare endocrine disorder that can be caused by mutations in the β-subunit of thyrotropin (TSHB). The TSHB mutation C105Vfs114X leads to isolated thyroid-stimulating-hormone-(TSH)-deficiency and results in a severe phenotype. The aim of this study was to gain more insight into the underlying molecular mechanism and the functional effects of this mutation based on two assumptions: a) the three-dimensional (3D) structure of TSH should be modified with the C105V substitution, and/or b) whether the C-terminal modifications lead to signaling differences. 2) Methods: wild-type (WT) and different mutants of hTSH were generated in human embryonic kidney 293 cells (HEK293 cells) and TSH preparations were used to stimulate thyrotropin receptor (TSHR) stably transfected into follicular thyroid cancer cells (FTC133-TSHR cells) and transiently transfected into HEK293 cells. Functional characterization was performed by determination of Gs, mitogen activated protein kinase (MAPK) and Gq/11 activation. 3) Results: The patient mutation C105Vfs114X and further designed TSH mutants diminished cyclic adenosine monophosphate (cAMP) signaling activity. Surprisingly, MAPK signaling for all mutants was comparable to WT, while none of the mutants induced PLC activation. 4) Conclusion: We characterized the patient mutation C105Vfs114X concerning different signaling pathways. We identified a strong decrease of cAMP signaling induction and speculate that this could, in combination with diverse signaling regarding the other pathways, accounting for the patient's severe phenotype

Anwendung von Organozinnhydridhalogeniden in der organischen Synthese

Dibutylzinnhydridhalogenide Bu2SnHX bieten eine Alternative zu dem häufig angewandten Tributylzinnhydrid. Da sie bereits bei Raumtemperatur unter Bildung von Stannylradikalen zerfallen, ermöglichen sie Radikalkettenreaktionen bei wesentlich milderen Bedingungen als bei der Verwendung von Tributylzinnhydrid, außerdem erweist sich die Abtrennung von zinnorganischen Nebenprodukten als wesentlich einfacher. Es wurden verschiedene Hydride Bu2SnHX in Reaktionen eingesetzt, bei denen zuvor Tributylzinnhydrid angewendet wurde, um Unterschiede in Ausbeute und Selektivität zu untersuchen. Es wurden erfolgreich Reduktionen von Halogen- und Carbonyl-Verbindungen durchgeführt sowie reduktive Cyclisierungen, Ringerweiterungen und reduktive Aminierungen. Als problematisch erwiesen sich intermolekulare C,C-Verknüpfungen (Giese-Reaktion) und die Barton-McCombie-Reaktion. Es hat sich gezeigt, daß die gemischten Hydride Bu2SnHX wesentlich reaktiver sind, auch bei tiefen Temperaturen werden hohe Umsätze erzielt. Die H-Übertragung erfolgt schneller als bei Einsatz von Bu3SnH. Es wurden Methoden zur Verwendung katalytischer Zinn-Mengen untersucht.Diorgano tinhydride halides Bu2SnHX are an alternative to the often used tributyltinhydride. As they are decomposing even at room temperature or below, they make free radical chain reactions possible under milder conditions than tributyltinhydride. Furthermore the removal of tin containing by-products is easier. Several mixed tin hydrides Bu2SnHX have been used in reactions, which have been carried out with tributyltinhydride before to investigate differences in yield and selectivity. Reductions of halogen- and carbonyl compounds have been performed successfully, as well as reductive cyclisations, ring expansions and reductive aminations. Intermolecular C,C-couplings (Giese-reaction) and the Barton-McCombie-reaction have been proved to be problematic. Mixed hydrides have been shown to be more reactive, high yields have been achieved even at low temperatures. The hydrogen-transfer is faster than in the case of tributyltinhydride. Methods have been investigated for the use of catalytic amounts of tin compounds

The COVID-19 Lockdown Effect on the Intention to Purchase Sustainable Brands

After the World Health Organization (WHO) declared the COVID-19 pandemic on 11 March 2020, almost all European countries entered a lockdown. This context caused sudden changes at multiple levels, affecting the way people were working, buying, studying and even the way they were interacting. Moreover, during lockdown people showed a special attention to local and sustainable brands giving momentum to the interest on sustainability, that has been increasing in the last years. Therefore, this study aims to determine the intention of buying sustainable and local brands due to the COVID-19 lockdown. An online survey was conducted for two groups of young adults from Spain and Romania, between April and June 2020. The questionnaire respects the methodological recommendations of Azjen (1985) and related literature on how to construct a survey based on the theory of planned behavior (TPB) and it aims to gather information about the three main constructs that determine the individual’s behavioral intention: attitudes, subjective norms and perceived behavioral control. The results suggest that both Spanish and Romanian samples intended to buy more local and sustainable brands, despite the slightly different attitudes. Moreover, the data show that both subjective norms and perceived behavioral control influence attitudes toward sustainable and local brands, and hence, indirectly the intention to buy sustainable products. The outcomes are adding to the literature on sustainability, and understanding the effects of COVID- 19 on consumer behavior. Additionally, the results can help better understand the importance of sustainability in Spain and Romania, and therefore, offering support to practitioners in building policies and programs that encourage a sustainable lifestyle.Economí

The competitive NMDA antagonist CPP protects substantia nigra neurons from MPTP-induced degeneration in primates

Degeneration of nigrostriatal dopaminergic neurons is the primary histopathological feature of Parkinson's disease. The neurotoxin MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) induces a neurological syndrome in man and non-human primates very similar to idiopathic Parkinson's disease by selectively destroying dopaminergic nigrostriatal neurons. This gives rise to the hypothesis that Parkinson's disease may be caused by endogenous or environmental toxins. Endogenous excitatory amino acids (EAAs) such as L-glutamate could be involved in neurodegenerative disorders including Parkinson's disease. We report in this study that the competitive NMDA antagonist CPP (3-((+/-)-2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid) protects nigral tyrosine hydroxylase (TH) positive neurons from degeneration induced by systemic treatment with MPTP in common marmosets. This indicates that EAAs are involved in the pathophysiological cascade of MPTP-induced neuronal cell death and that EAA antagonists may offer a neuroprotective therapy for Parkinson's disease

Abgeformte polymere Mikrostrukturen für die optische Informationstechnik

Für eine stärkere Verbreitung der optischen Informationstechnik werden optische Wellenleiterkomponenten benötigt, die u.a. die Signale optisch schalten und aufsplitten können. Dazu ist die sogenannte SIGA-Technik entwickelt worden, mit der polymere, integriert optische Mikrostrukturen mittels kostengünstiger Replikationstechniken hergestellt werden können.In dieser Arbeit wird dabei auf die Replikationstechniken im Rahmen der SIGA-Technik näher eingegangen. Dazu werden die beiden eingesetzten Abformverfahren des Spritzgießens und Heißprägens dargestellt, mit den Randbedingungen für Mikrostrukturen im allgemeinen und integriert optische Mikrostrukturen im speziellen.Das für integriert optische Mikrostrukturen entwickelte modifizierte Mikrospritzprägen wird ausführlich erläutert, wobei auf die speziellen Werkzeuge, das verwendete Equipment und die damit durchgeführten Prozesse näher eingegangen wird.Das zweite Abformverfahren, das Heißprägen, wird hauptsächlich als Laboranwendung für Prototypen eingesetzt, aber zunehmend auch bei bestimmten empfindlichen Strukturen und besonderen Geometrien, dargestellt am Beispiel einer optischen Rückwand, die über ihre gesamte Länge (10 bis 50 Zentimeter) integriert optische Mikrostrukturen enthält.Ausführlich wird auf Optimierungsmöglichkeiten für die Abformung von Mikrostrukturen eingegangen.Anwendungsbeispiele, vornehmlich aus dem Bereich der optischen Informationstechnik, aber auch aus der Mikrofluidik, demonstrieren das Einsatzpotential der Mikroabformtechnik.To ensure a wider spreading of optical IT optical waveguides are necessary to switch and split optical signals. Therefore the so called SIGA-technique has been developed for manufacturing polymeric, integrated optical microstructures with low-cost replication techniques.Within this thesis the attention is directed towards the replication techniques as part of the SIGA technique. The two used moulding techniques as injection moulding and hot embossing are described putting up marginal conditions for microstructures in general and integrated optical microstructures in particular.The modified micro injection moulding developed especially for integrated optical microstructures is explained in detail. Close attention is given to the special tools, the used equipment and the processes carried out.The second moulding technique, hot embossing, is usually applied as a lab technique for prototypes, but more and more for difficult structures with special geometries, too, shown by the example of an optical backplane. This backplane, which is several centimetres long (10 to 50 cm), has integrated optical microstructures over its complete length.The optimization for replication of microstructures is described in detail.Application examples, especially from optical information technique, but also from micro fluidics, demonstrate the high potential of micro moulding / micro replication techniques

Deliver Smart, Not More! Building Economically Sustainable Competitiveness on the Ground of High Agri-Food Trade Specialization in the EU

Competitiveness has always been a multifaceted illusive concept, which has made it a real challenge for scholars and practitioners to find the most suitable measurement tools to completely encapsulate all the complex nuances of competitiveness. This becomes even more of a challenge when approached in relation to particular economic sectors. The agri-food sector is no exception, especially when considering all its interconnections with the other sectors: water, energy, transport, waste. All of them impact the achievement of the Sustainable Development Goals (SDGs). Similarly, scholars have been debating the meaning of sustainability for decades, some even arguing that it is a political, subjective, and, in some cases, self-contradictory concept. As far as the sustainability of agricultural competitiveness is concerned, the literature is still developing. It is much more focused on fostering environmental competitiveness, and less attention was paid to the strategies designed to capitalize on sustainable economic competitiveness—a concept that has attracted divergent opinions in the literature, mainly due to ambiguity. Thus, instead of falling into the pitfall of vagueness, this paper was aimed at bringing its contribution to this field by undertaking the research objective of exploring a single facet of sustainable agricultural competitiveness: the economic facet. Hence, this paper proposes the construction of the sustainable economic competitiveness index (SECI) with direct application for agri-food value chains. It consists of three attributes: (a) factor endowments, resource independence; (b) agricultural chain performance; and (c) national agricultural chain strategies and policies. In this study, SECI was tested against the cereal chain for a selection of EU countries, based on the data taken over from FAOSTAT and INTRACEN Trade Map, in the case of the 2011–2020 period. Various statistical and econometric methods were used to test the robustness of SECI. Results stand as proof that building sustainable agricultural economic competitiveness relies on a mix of strategic actions. The key vector in this mix is that trade flow patterns and policies must be calibrated in accordance with national factor endowments in order to achieve high levels of SECI. To add more managerial implications, this paper argues for the smart delivery of agri-food products with high added value instead of focusing on exporting big volumes of raw agricultural materials with little added value.The APC was funded by the authors. Juan Sapena acknowledges the financial supportof Grant PID2020-114646RB-C42 funded by MCIN-AEI/10.13039/501100011033. Juan Sapena andAndreea Apetrei are also indebted to the “Christeyns” Chair for Sustainable Economic Developmentfor its specific funding of their research.Economí

Klinische und molekularbiologische Effekte der Mutation C105Vfs114X im TSHB Gen

Kongenitale Hypothyreosen sind mit einer Inzidenz von etwa 1:3.500 die häufigste angeborene endokrine Erkrankung. Die meisten Patienten weisen eine primäre Hypothyreose auf, die in der Regel durch eine abnorme Entwicklung der Schilddrüse bedingt ist. Sekundäre beziehungsweise zentrale Hypothyreosen sind mit einer Inzidenz von ca. 1:50.000 wesentlich seltener. Eine der Ursachen dafür sind Mutationen im TSHB Gen, welche zu einem isolierten TSH-Mangel führen. Man ging lange davon aus, dass sekundäre Hypothyreosen zu einem wesentlich milderen Phänotyp als primäre Hypothyreosen führen. Dies trifft jedoch nicht für Patienten mit einer Mutation in der β-Untereinheit des TSH und insbesondere nicht für Patienten mit der Mutation C105Vfs114X zu. Hinzu kommt, dass diese Patienten nicht im Neugeborenscreening detektiert werden, da dieses auf der alleinigen Messung von TSH beruht, welches in diesen Fällen in der Regel unterhalb der Nachweisgrenze oder im unteren Normbereich liegt. Patienten mit dieser Mutation fallen mit ausgeprägten Symptomen einer schweren konnatalen Hypothyreose auf und haben langfristige psychomotorische sowie neurokognitive Defizite, da die Mutation meist nicht so früh erkannt wird, dass die Substitutionstherapie mit L-Thyroxin Langzeitschäden verhindert. Weshalb diese Mutation zu einem so schweren Phänotyp führt, sollte in dieser Arbeit untersucht werden. Ein besonderer Pathomechanismus, z.B. im Sinne eines invers-agonistischen Effekts des mutierten TSH am TSH-Rezeptor, wurde seit Ende der 90er Jahren vermutet. Besonderes Augenmerk wurde in dieser Arbeit auf den Verlust einer Disulfidbrückenbindung und den verkürzten C-Terminus gelegt. Dadurch potentiell bedingte Veränderungen der Formation des Proteins könnten diesen spefizischen Pathomechanismus bedingen. Um dies zu untersuchen, wurden in dieser Arbeit verschiedene TSHB-Mutanten generiert und rekombinantes hTSH durch transiente Transfektion in HEK293 Zellen hergestellt. Die Menge des hergestellten hTSH wurde gemessen und das hTSH funktionell charakterisiert. In dieser Arbeit konnte ein invers-agonistischer Effekt der Mutation ausgeschlossen werden. Darüberhinaus zeigten die Experimente, dass der verkürzte C-Terminus nicht ursächlich für die Effekte der Mutation verantwortlich ist. Es ist nicht gelungen eine andere molekulare Ursache für diesen schwerwiegenden Phänotyp zu ermitteln. Dafür bedarf es weiterführender Experimente mit zusätzlichen Mutanten, um weitere Erkenntnisse zu den Auswirkungen dieser Mutation zu erlangen. Ein tieferes Verständnis dieser Mutation könnte nicht nur dazu beitragen, auch Patienten und ihren Eltern das Krankheitsbild schlüssig zu erklären, sondern ggf. auch dazu beitragen, neue zielgerichtete Therapien z.B. für Patienten mit Hyperthyreose zu entwickeln. Nicht zuletzt kann eine auf molekularer Ebene gesicherte Erkenntnis einer schweren Beeinträchtigung der Schilddrüsenhormonbiosynthese dazu beitragen, dass die Strategie des Neugeborenenscreenings auf angeborene Hypothyreose revidiert wird.Congenital hypothyroidism is – with an incidence of approximately 1:3,500 – the most common congenital endocrine disorder. Most patients are diagnosed with primary congenital hypothyroidism, which is in most cases caused by a developmental defect of the thyroid gland. Comparably, secondary (central hypothyroidism) is very rare with an incidence of 1:50,000. One of the reasons for central hypothyroidism are mutations in the TSHB gene, which lead to an isolated TSH-deficiency. It has long been assumed that central hypothyroidism leads to a milder phenotype than primary hypothyroidism does. However, this does not apply to mutations in the β-subunit, especially not to the mutation C105Vfs114X. Furthermore, these patients are not detected through newborn screening as this screening is based on TSH, in which β-subunit mutations result in TSH concentrations below or at the lower end of the reference range. Affected patients therefore present with severe signs of congenital hypothyroidism and with long-term psychomotor and neurocognitive deficiencies if the diagnosis is not made in the neonatal period and the start of l-thyroxine replacement is delayed. The aim of this study was to investigate the mechanism by which this specific mutation leads to a more severe phenotype than other forms of central hypothyroidism. Since the 1990s it was presumed that an inverse-agonistic effect of the mutated TSH at the receptor could cause this phenotype. This again, could be caused by the shortened c-terminus, an altered amino acid sequence or the loss of one of the disulfide bridges, which could lead to a modified formation of the protein. Different mutants were generated for the experiments; recombinant hTSH was produced using transient transfection in HEK293 cells. The produced hTSH was measured and a functional characterization of the wildtype as well as of the mutants was conducted. With these experiments an inverse-agonistic effect of the mutation was ruled out and it was proved that the shortened c-terminus is not responsible for the effects of the mutation. However, it was not possible to identify the exact molecular mechanism. In order to do so, further experiments with additional variants will be required. Besides an explanation for patients and their families, a deeper understanding of this mutation could help to develop novel targeted therapies especially for patients with hyperthyroidism. Not least, molecular evidence for a severely altered production of thyroid hormones would underline the necessity to revise the strategy of newborn screening programs for congenital hypothyroidism

Linguistic stress, within-word position, and grammatical class in relation to early childhood stuttering

Abstract. The purpose of the present study was to investigate whether the link that has been established between stuttering and linguistic stress in adolescents and adults (the so-called stress effect) can also be observed in childhood stuttering. To account for confounding variables, both within-word position and grammatical class were measured, because these factors covary with linguistic stress. Speech samples of twenty-two preschool children (mean time of 9 months since onset of stuttering) were analyzed. The relative stress of each syllable was rated and syllables were categorized into long and short stressed, unstressed and intermediately stressed syllables. Results showed that 98 % of stuttering events occurred on first syllables of words, i.e. a clear word-initial effect. Stuttering frequency on first syllables of function words was 16.9 % and significantly higher than the frequency of stuttered first syllables of content words (11.5 %). In function words short stressed syllables and intermediately stressed syllables were stuttered more often than unstressed syllables. The analysis for individual disfluency types revealed that, for function words, stuttering on short stressed syllables was associated with prolongations and syllable repetitions. However, in intermediately stressed syllables stuttering coincided most often with one-syllable word repetitions. This differentiation of the stress effect may suggest different causal mechanisms underlying these disfluency types

Rift Valley fever phlebovirus NSs protein core domain structure suggests molecular basis for nuclear filaments

Rift Valley fever phlebovirus (RVFV) is a clinically and economically important pathogen increasingly likely to cause widespread epidemics. RVFV virulence depends on the interferon antagonist non-structural protein (NSs), which remains poorly characterized. We identified a stable core domain of RVFV NSs (residues 83-248), and solved its crystal structure, a novel all-helical fold organized into highly ordered fibrils. A hallmark of RVFV pathology is NSs filament formation in infected cell nuclei. Recombinant virus encoding the NSs core domain induced intranuclear filaments, suggesting it contains all essential determinants for nuclear translocation and filament formation. Mutations of key crystal fibril interface residues in viruses encoding full-length NSs completely abrogated intranuclear filament formation in infected cells. We propose the fibrillar arrangement of the NSs core domain in crystals reveals the molecular basis of assembly of this key virulence factor in cell nuclei. Our findings have important implications for fundamental understanding of RVFV virulence.Publisher PDFPeer reviewe