The biogenesis of alkaloids. XX. The induced biogenesis of stachydrine.

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Kids or courses? Gender differences in the effects of active labor market policies

We investigate active labor market programs in Austria. We find only small effects, if any, for most of the programs. However, the programs may have unintended consequences for women. In particular for younger women, a key effect of the programs and one reason for the male-female effect differential that is observed in the literature is to reduce or postpone pregnancies and to increase their attachment to the labor force. Furthermore, the variables capturing pregnancies and times of parental leave play a key role in removing selection bia

Good Manufacturing and Hygienic Practices at Small and Medium Scale Pineapple Processing Enterprises in Rwanda

The purpose of this study was to assess the compliance of small and medium scale pineapple processing enterprises in Rwanda to the Good Manufacturing Practices (GMP) and Good Hygiene Practices (GHP) as set by the national, regional and Codex Alimentarius Commission (CAC) food safety and hygiene regulations. The layout design and practices of hygiene for 16 small and 11 medium pineapple processing enterprises were characterised by conducting face-to-face interviews using questionnaires and observations. None of the visited enterprises had clear criteria of the delivery procedure of raw pineapples and none had a signed contract with the suppliers and this led to uncontrolled primary production. The plant layout and design of both small and medium enterprises were characterised by insufficient number of working rooms and there was  crisscrossing in the production line of 81.8% and 87.5% respectively for medium and small enterprises. The clean and unclean sectors were only separated at 63.6% and 31.2% of the medium and small enterprises respectively. In generally, the status of all enterprise categories, their surrounding areas and equipment was classified as unsatisfactory. The majority of enterprises did not have temperature and time control systems and for both enterprise categories, there was no clear plan for waste disposal, plus pest and insect control. Lack of compliance to the standard requirements was observed in the majority of both enterprise categories. In order for these enterprises to access export market, it is recommended that training on the requirements for GMP and HGP for food processing industries by the Rwanda Bureau of Standards and other government and non-government institutions be provided. Training programs on basic food safety and hygiene in food processing enterprises need to be in place to support implementation of prerequisite programs in the small scale fruit processing enterprises. Keywords: Good manufacturing practices, Small and medium enterprises, Pineapple processing, Standard

Issues Affecting Small- and Medium-Scale Pineapple Processing Enterprises in Rwanda: A Cross-sectional Study

Fruit processing small- and medium-scale enterprises (SMEs) play a vital role in the national economic development of Rwanda. Though they receive attention from the government, they still face numerous constraints that hinder their development. A cross-sectional study design was used to profile 16 small-scale and 11 medium-scale pineapple-processing enterprises in the country and assess the issues affecting them. From July to August in 2011, data was collected using a structured questionnaire and face-to-face interviews with enterprise managers. Results showed that most managers of these SMEs are male, married, within the age range of 36–40 years, have had formal education, and works 10 h per day. Most of these SMEs are located in the Eastern Province near pineapple production areas. The mean for the capital investment for small-scale processors was estimated at US$11,066 and for medium enterprises at US$805,000. Majority of small-scale enterprises had 10 employees or less while majority of the medium-scale enterprises had more than 10 employees, with both enterprises hiring mostly unskilled laborers working 8 h per day. Results reveal that the major constraints in pineapple-processing industry in Rwanda were the following: access to modern processing equipment and proper packaging material, access to raw materials, high cost of water and electricity, high transportation cost, and limited knowledge and skills of human resources. This article provides various recommendations on how SMEs can overcome these constraints and produce quality products that can penetrate the export market

Wechselwirkungen zwischen Chromophor und Proteinmatrix in den autofluoreszierenden Proteinen GFP und drFP583 (DsRed)

Durch die genetische Fusion mit dem Grün Fluoreszierenden Protein können seit 1994 Proteine oder Genexpression in vivo visualisiert werden, ein bedeutender Fortschritt für die Lebenswissenschaften. Die Nutzungsmöglichkeiten können z. B. mit der gleichzeitigen Verwendung von fluoreszenzspektroskopisch unterscheidbaren GFP-Varianten (multispektrale Detektion) erweitert werden. Eine Veränderung der spektralen Eigenschaften des Proteins unter Beibehaltung seiner Funktionalität in vivo kann nur durch Mutagenese erfolgen. Das Ziel dieser Arbeit ist es, einen Beitrag zur grundlegenden Charakterisierung der Wechselwirkungen zwischen dem Chromophor und der umgebenden Proteinmatrix zu leisten. Durch die Substitution von T203 in wt-GFP kann gezeigt werden, dass die B-Spezies gegenüber der A- und der I-Spezies durch eine Wasserstoff-Brücke von T203 zum Chromophor gekennzeichnet ist. Diese bewirkt gegenüber der I-Spezies ein Blauverschiebung der Absorption um ca. 23 nm. Die I-Spezies kann durch Doppelmutationen stabilisiert werden und stellt unter den GFP-Varianten einen neuen spektralen Phänotyp dar. Die Tendenz zur Dimerisierung ermöglicht die absorptionsspektroskopische Darstellung einer Population der I-Spezies in wt-GFP. Gleichzeitig wird das Dimerisierungsmodell experimentell bestätigt und mit einer relativ schwachen Dimerisierungskonstante von ca. 3 mM-1 ergänzt. Die spektroskopischen Eigenschaften wichtiger GFP-Varianten lassen sich nun mit einem molekularen Modell erklären. So wird bewiesen, dass die Absorptionsrotverschiebung um 33 nm durch die Mutation T203Y (YFP-Varianten) nur zu ca. 1/3 auf spezifisch aromatischen Wechselwirkungen beruht. Zusammen mit den EGFP-Varianten eignet sich die T203V-Variante für eine neue Art der multispektralen Detektion auf der Grundlage der sehr gut separierten Anregungsspektren. Die Kenntnisse über die Vorgänge bei der Dimerisierung von wt-GFP bzw. GFPuv lassen sich prinzipiell für die intrazelluläre und lokale in vivo-Messung effektiver Proteinkonzentrationen bzw. von Protein-Protein-Wechselwirkungen nutzen. Durch kombinatorische Mutagenese wurde deutlich, dass der schnelle Protonentransfer (ESPT) nach Anregung der A-Spezies das negativ geladene E222 als Akzeptor benötigt. Ansonsten wird der Prozess deutlich verlangsamt, was mit einer entsprechenden Abnahme der Quantenausbeute für die grüne Emission einhergeht. In der Variante S65G/T203V/E222Q wird zusätzlich der strahlunglose Verlust der Anregungsenergie von A* stark verringert. Dadurch dominiert die blaue A*-Emission das Emissionsspektrum. Dieser neue spektrale GFP-Phänotyp ist für multispektrale Anwendungen geeignet. Von den GFP-homologen Proteinen, die erst seit Ende 1999 entdeckt werden, hat das stark rot emittierende aus Discosoma (DsRed) die langwelligste Emission und ist deshalb für in vivo Applikationen am interessantesten. Es wird bewiesen, dass der Chromophor des Proteins über die langsame kovalente Modifikation des intermediär entstehenden GFP-Chromophors gebildet wird. Mit der Entfaltung des Proteins wird dieser Prozess teilweise revertiert. Aus einer Proteinbibliothek konnten Varianten identifiziert werden, in denen räumlich gruppierte Einzelmutationen diesen finalen Maturierungsschritt in verschiedenem Ausmaß unterdrücken. Die Emission des intermediären GFP-Chromophors kann von dem finalen DsRed-Chromophor absorbiert werden (FRET), was auf eine starke Oligomerisierung hindeutet

How can AI regulation be effectively enforced? : comparing compliance mechanisms for AI regulation with a multiple-criteria decision analysis

Award date: 17 June 2022. Supervisor: Professor Andrea Renda (European University Institute)Newly emerging AI regulations need effective and innovative enforcement and compliance mechanisms to assure that fundamental and human rights are protected when using an AI system. This study compares four different compliance mechanisms namely ‘Real-Time and Automated Conformity Assessment’, ‘Standardization and Certification’, ‘Algorithmic Impact Assessment’ and ‘Algorithmic Auditing’ as well as three different assurers of compliance namely deployers, notified bodies and civil society organisations. With an MCDA, this research has shown that civil society-based compliance mechanisms are believed to be less effective, less feasible and more costly compared to all other compliance mechanisms. Second, external compliance mechanisms (by notified bodies) were rated to be more effective but also more difficult to implement compared to internal compliance mechanisms. Third, algorithmic auditing scored highest among all policy options. Fourth, despite its experimental nature, automated and real-time compliance mechanisms are not scored significantly lower than other compliance mechanisms

Human rhinovirus infection up-regulates MMP-9 production in airway epithelial cells via NF-{kappa}B

Human rhinovirus (HRV) infections up-regulate proinflammatory mediators and growth factors that are associated with exacerbations of inflammatory airway diseases, such as asthma and chronic obstructive pulmonary disease (COPD). Matrix metalloproteinase (MMP)-9 was shown to be increased in the airways of patients with asthma and COPD. We sought to determine whether HRV infection modulated the expression of MMP-9 and its highest-affinity inhibitor, the tissue inhibitor of metalloproteinase (TIMP)-1, and we explored the mechanism by which this modulation occurs. In vitro studies, using RT-PCR, ELISA, zymography, and a fluorescent activity assay, demonstrated that MMP-9 mRNA, protein, and activity were increased upon infection with HRV, whereas TIMP-1 mRNA and protein remained unchanged. These results were verified in vivo, using nasal lavage samples obtained from subjects with confirmed rhinovirus infections. Human rhinovirus infections were shown to up-regulate NF-kappaB, and NF-kappaB has also been reported to play a role in the expression of MMP-9. We therefore investigated the role of NF-kappaB in HRV-induced MMP-9 expression. Using two inhibitors of IkappaBalpha kinase beta, we observed a concentration-dependent decrease in HRV-induced MMP-9 expression. The role of NF-kappaB in HRV-induced MMP-9 expression was further confirmed using MMP-9 promoter luciferase constructs, which demonstrated that an NF-kappaB site at -620/-607 base pairs was necessary for HRV-induced MMP-9 expression. Electrophoretic mobility shift assays and supershift assays confirmed the nuclear translocation and binding of p50/p65 NF-kappaB subunits to an MMP-9-specific NF-kappaB oligonucleotide. This increase in MMP-9 may be a mechanism by which rhinovirus infections contribute to airway inflammation and, potentially, to airway remodeling

Molecular strategies for increasing seed oil content in canola

xix, 245 leaves : ill. ; 28 cm.Previous research has shown that microsomal DGAT activity from cultures of Brassica napus was stimulated by human acylation stimulating protein (ASP) and bovine serum albumin (BSA). Genetic constructs were engineered to facilitate the expression of ASP or BSA in developing seeds to test the stimulatory effect of these proteins at the site of TAG formation. As well, genetic constructs were designed to produce a truncation of the BSA polypeptide in an attempt to localize the portion fo the macromolecule responsible for stimulation of DGAT activity. An oleosin promoter was used for seed specific expression and to express the polypeptides at a precisely cooridnated time when oil was accumulating in the developing seeds. Lipid analysis coordinated time when oil was accumlating in the developing seeds. Lipid analysis was performed on the seeds of transgenic plants designed to cytosolically express these mammalian proteins and the seeds of control plants. The first generation data revealed that the total lipid within the mature seeds of ASP and BSA plants was not significantly different from the total lipid of negative control plants using both gravimetric and low resolution-nuclear magnetic resonance methods of analysis. The seeds from ASP 8 and ASP 10 plants, however, did produce significantly more lipid on a per seed basis as compared to negative control plants. The levels of the fatty acid composition for total acyl lipids were measured in these first generation transgenic plants. ASP 3 had significantly lower levels of linoleic acid, ASP 14 had significantly lower levels of a-linolenic acid and BSA 11 had significantly higher levels of both of these fatty acids in comparison to negative control plants