Development aid, one of the most important mechanisms for the redistribution of global wealth, represents financial flows with economic growth and social improvement as their primary objective. ...Recently, international development cooperation has been in the process of change that requires the involvement of new actors, the use of new instruments, and expanding the fields of work. In this article, the authors introduce the Slovenian model of international development cooperation, emphasising the criteria for deciding the direction of development cooperation. The criteria are clear interest of the beneficiary state, content priority fields, and geographical priority areas. The authors also discuss Slovenia's active collaboration with the United Nations Industrial Development Organization in international development cooperation. Special attention is given to three successful development projects in Africa that Slovenia recently financed. All three selected projects are based on principles of sustainable development and were realized in Kenya, Tanzania, and Uganda.
V nalogi smo v teoretičnem delu predstavili teoretične novosti o storitvah, razvoj in pomen mednarodne trgovine s storitvami (v primerjavi z izdelki), proučili lastnosti in značilnosti storitev in ...mednarodne trgovine z njimi, zaradi katerih se razlikujejo od lastnosti in značilnosti dejavnosti izdelkov (dobrin, proizvodov, produktov).V povezavi s hitrim razvojem mednarodne trgovine s storitvami smo proučili značilnosti in lastnosti inovacijskih dejavnosti pri storitvah in kaj jih loči od inovacijskih dejavnosti v proizvodnih dejavnostih. Na podlagi različnih virov in lastnega proučevanja smo sestavili seznam 13 lastnosti in značilnosti storitev, ki so v tej doktorski nalogi prvič zbrane skupaj, na enem mestu. Te lastnosti natančno določajo storitve in (mednarodno) trgovino s storitvami ter kaj jih loči od lastnosti izdelkov in (mednarodne) trgovine z njimi.V povezavi s storitvami smo proučili elemente gibljivost kapitala, gibljivost dela (ljudi), nadnacionalna podjetja, uporabo in razvoj novih tehnologij, čezmejni pretok informacij, ki vplivajo na mednarodno trgovino s storitvami in ovire v tej trgovini.Podali smo nekaj osnovnih statističnih podatkov o pomenu storitev in storitvenih dejavnosti v mednarodni trgovini. Proučili smo inovacije in inovacijske dejavnosti v storitvenih dejavnostih. Pri tem smo se v analitičnem delu strukturno omejili na poslovne storitve kot ožji del področja nefinančnih storitvenih dejavnosti, geografsko pa na območje Slovenije.V nadaljevanju smo preučili teoretična izhodišča o ekonomskem vplivu inovacijskih dejavnosti na področju storitvenih dejavnosti na obseg mednarodne trgovine s storitvami (izvoz) in obratno. V izbranem obdobju 2010-2016 smo temu dodali še teoretična izhodišča o ekonomskem vplivu investicijskih dejavnosti v razvoj na področju storitvenih dejavnosti na obseg mednarodne trgovine s storitvami (izvoz).V jedrnem delu disertacije smo oblikovali raziskovalni model, ki temelji na uporabi analitičnih metod, ki so jih uporabljali raziskovalci na področjih proizvodnih dejavnosti in smo jih aplicirali na področju poslovnih storitvenih dejavnosti.Za ugotavljanje ekonomskih učinkov izvoznih dejavnosti z vidika vpliva inovacijskih dejavnosti in investicijskih aktivnosti v razvoj v sektorju dejavnosti poslovnih storitev v Sloveniji je bilo potrebno zgraditi model v dveh korakih. Zato smo predstavili klasični model rasti in razvoj modela endogene rasti, ki je predstavljal osnovo za oblikovanje našega modela. Najprej smo uporabili enostaven "probit" model, s katerim smo ugotavljali vpliv izvoznih dejavnosti na gospodarstvo ob pogoju danih inovacijskih dejavnosti (za izbrani obdobji 2002-2008 in 2010-2016) in vpliv izvoznih dejavnosti na gospodarstvo ob pogoju danih investicijskih dejavnosti (za opazovano obdobje 2010-2016).V drugem koraku smo model posplošili in uporabili inovacijske (investicijske) dejavnosti kot endogene spremenljivke z uporabo sočasnega (simultanega) "probit" modela.Rezultati meritev so pokazali, da so inovacijske dejavnosti v podjetjih v sektorju poslovnih storitev pomembne za uspeh na tujih trgih. Podjetja z novimi ali izboljšanimi storitvami so bolje konkurirala na tujih trgih kot neinovativna storitvena podjetja. Med letoma 2010 in 2016 so rezultati pokazali verjetnosti, da bi skoraj vsako drugo podjetje, ki je inovacijsko intenzivno, postalo tudi izvozno intenzivno. Zaradi visokega tveganja pri ocenjevanju modela pa ni bilo mogoče potrditi obratnega vpliva, da izvozne dejavnosti pozitivno vplivajo na inovacijske dejavnosti.Rezultati meritev so potrdili domnevo, da so investicijske dejavnosti v razvoj pozitivno vplivale na večje izvozne dejavnosti v podjetjih za merjeno obdobje med letoma 2010 in 2016. Bolj kot so podjetja investirala v razvoj, večja je bila verjetnost, da bodo izvozniki.Za merjeno obdobje med letoma 2010 in 2016 so izračuni pokazali verjetnost pozitivnega vpliva izvoznih na investicijske intenzivnosti.
The G4C2 hexanucleotide repeat expansion mutation (HREM) in C9ORF72, represents the most common mutation associated with amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration ...(FTLD). Three main disease mechanisms have been proposed to date: C9ORF72 haploinsufficiency, RNA toxicity, and accumulation of dipeptide repeat proteins. Pure GC content of the HREM potentially enables the formation of various non-B DNA structures such as G-quadruplexes and i-motifs. These structures are proposed to act as promoters and regulatory elements affecting replication, transcription and translation of the surrounding region. G-quadruplexes have already been shown on the G-rich sense DNA and RNA strands (G4C2)n, the structure of the anti-sense (G2C4)n strand remains unresolved. Similar C-rich sequences may, under acidic conditions, form i-motifs consisting of two parallel duplexes in a head to tail orientation held together by hemi-protonated C(+)-C pairs. We show that d(G2C4)n repeats do form i-motif and protonated hairpins even under near-physiological conditions. Rather than forming a DNA duplex, i-motifs persist even in the presence of the sense strand. This preferential formation of G-quadruplex and i-motif/hairpin structures over duplex DNA, may explain HREM replicational and transcriptional instability. Furthermore, i-motifs/hairpins can represent a novel pharmacological target for C9ORF72 associated ALS and FTLD.
The hexanucleotide expansion GGGGCC located in
gene represents the most common genetic cause of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar dementia (FTLD). Since the discovery one ...of the non-exclusive mechanisms of expanded hexanucleotide G
C
repeats involved in ALS and FTLD is RNA toxicity, which involves accumulation of pathological sense and antisense RNA transcripts. Formed RNA foci sequester RNA-binding proteins, causing their mislocalization and, thus, diminishing their biological function. Therefore, structures adopted by pathological RNA transcripts could have a key role in pathogenesis of ALS and FTLD. Utilizing NMR spectroscopy and complementary methods, we examined structures adopted by both guanine-rich sense and cytosine-rich antisense RNA oligonucleotides with four hexanucleotide repeats. While both oligonucleotides tend to form dimers and hairpins, the equilibrium of these structures differs with antisense oligonucleotide being more sensitive to changes in pH and sense oligonucleotide to temperature. In the presence of K
ions, guanine-rich sense RNA oligonucleotide also adopts secondary structures called G-quadruplexes. Here, we also observed, for the first time, that antisense RNA oligonucleotide forms i-motifs under specific conditions. Moreover, simultaneous presence of sense and antisense RNA oligonucleotides promotes formation of heterodimer. Studied structural diversity of sense and antisense RNA transcripts not only further depicts the complex nature of neurodegenerative diseases but also reveals potential targets for drug design in treatment of ALS and FTLD.
The G4C2 hexanucleotide repeat expansion, located in the first intron of the C9ORF72 gene, represents a major genetic hallmark of amyotrophic lateral sclerosis and frontotemporal lobar degeneration. ...Several hypotheses have been proposed on how the transcribed repeat RNA leads to the development of amyotrophic lateral sclerosis and frontotemporal lobar degeneration. However, despite their importance, factors affecting the transcription of expanded-repeat RNA are not well known. As transcription is dependent on the DNA containing the expanded repeats, it is crucial to understand its structure. G-quadruplexes are known to affect expression on the level of DNA, therefore whether they form on the expanded-repeat DNA constitutes an important biological question. Using nuclear magnetic resonance and circular dichroism spectroscopy we show that DNA G4C2 with varying number of repeats d(G4C2)n form planar guanine quartets characteristic of G-quadruplexes. Additionally, we show DNA G-quadruplexes can form inter- and intra-molecularly in either parallel or anti-parallel orientation, based on d(G4C2) sequence length. This potential structural heterogeneity of longer disease-relevant repeats should therefore be taken into account when studying their role in disease pathogenesis.
Genetic studies have managed to explain many cases of familial amyotrophic lateral sclerosis (ALS) through mutations in several genes. However, the cause of a majority of sporadic cases remains ...unknown. Recently, epigenetics, especially miRNA studies, show some promising aspects. We aimed to evaluate the differential expression of 10 miRNAs, including miR-9, miR-338, miR-638, miR-663a, miR-124a, miR-143, miR-451a, miR-132, miR-206, and let-7b, for which some connection to ALS was shown previously in ALS culture cells, animal models or patients, and in three miRNA host genes, including
(miR-9),
(miR-338), and
(miR-638), in leukocyte samples of 84 patients with sporadic ALS. We observed significant aberrant dysregulation across our patient cohort for miR-124a, miR-206, miR-9, let-7b, and miR-638. Since we did not use neurological controls we cannot rule out that the revealed differences in expression of investigated miRNAs are specific for ALS. Nevertheless, the group of these five miRNAs is worth of additional research in leukocytes of larger cohorts from different populations in order to verify their potential association to ALS disease. We also detected a significant up-regulation of the
gene and down-regulation of the
gene, and thus, for the first time, we connected these with sporadic ALS cases. These findings open up new research toward miRNAs as diagnostic biomarkers and epigenetic processes involved in ALS. The detected significant deregulation of
and
in sporadic ALS also represents an interesting finding. The
gene was previously found to be mutated in Charcot-Marie-Tooth neuropathy-type CMT2M and centronuclear myopathy (CNM). In addition, as recent studies connected
and frontotemporal dementia (FTD) and
and hereditary spastic paraplegia (HSP), these two genes together with our results genetically connect, at least in part, five diseases, including FTD, HSP, Charcot-Marie-Tooth (type CMT2M), CNM, and ALS, thus opening future research toward a better understanding of the cell biology involved in these partly overlapping pathologies.
Amyotrophic lateral sclerosis is a late-onset disorder primarily affecting motor neurons and leading to progressive and lethal skeletal muscle atrophy. Small RNAs, including microRNAs (miRNAs), can ...serve as important regulators of gene expression and can act both globally and in a tissue-/cell-type-specific manner. In muscle, miRNAs called myomiRs govern important processes and are deregulated in various disorders. Several myomiRs have shown promise for therapeutic use in cellular and animal models of ALS; however, the exact miRNA species differentially expressed in muscle tissue of ALS patients remain unknown. Following small RNA-Seq, we compared the expression of small RNAs in muscle tissue of ALS patients and healthy age-matched controls. The identified snoRNAs, mtRNAs and other small RNAs provide possible molecular links between insulin signaling and ALS. Furthermore, the identified miRNAs are predicted to target proteins that are involved in both normal processes and various muscle disorders and indicate muscle tissue is undergoing active reinnervation/compensatory attempts thus providing targets for further research and therapy development in ALS.
Most Varroa induced colony losses occur during the autumn or winter season as a consequence of an elevated Varroa infestation level and an insufficient health status of the adult bees. Even with an ...initial low Varroa infestation in early spring, critical mite and virus infection levels can be reached before winter if colonies continuously rear brood throughout the whole season. To overcome this challenge, beekeepers can artificially interrupt brood production by suitable management procedures, depending on their type of beekeeping operation. To assess their efficacy, associated workload, and impact on colony development, different methods for brood interruption (queen caging with the combination of oxalic acid treatment, total brood removal, trapping comb technique) were tested during two seasons in 11 locations on 370 colonies in 10 European countries. A protocol was developed to standardize the methods' application across different environmental conditions. The efficacy of queen caging depended on the mode of oxalic acid application and ranged from 48.16% to 89.57% mite removal. The highest efficacies were achieved with trickling a 4.2% solution (89.57%) and with the sublimation of 2 g of oxalic acid (average of 88.25%) in the broodless period. The efficacy of the purely biotechnical, chemical-free trapping comb and brood removal methods did not differ significantly from the queen caging groups. We conclude that a proper application of one of the described brood interruption methods can significantly contribute to an efficient Varroa control and to the production of honey bee products meeting the highest quality and food-safety standards.
Repeat expansions in the C9orf72 gene are a common cause of amyotrophic lateral sclerosis and frontotemporal lobar degeneration, two devastating neurodegenerative disorders. One of the proposed ...mechanisms of GGGGCC repeat expansion is their translation into non-canonical dipeptide repeats, which can then accumulate as aggregates and contribute to these pathologies. There are five different dipeptide repeat proteins (polyGA, polyGR, polyPR, polyPA and polyGP), some of which are known to be neurotoxic. In the present study, we used BioID2 proximity labelling to identify the interactomes of all five dipeptide repeat proteins consisting of 125 repeats each. We identified 113 interacting partners for polyGR, 90 for polyGA, 106 for polyPR, 25 for polyPA and 27 for polyGP. Gene Ontology enrichment analysis of the proteomic data revealed that these target interaction partners are involved in a variety of functions, including protein translation, signal transduction pathways, protein catabolic processes, amide metabolic processes and RNA-binding. Using autopsy brain tissue from patients with C9orf72 expansion complemented with cell culture analysis, we evaluated the interactions between polyGA and valosin containing protein (VCP). Functional analysis of this interaction revealed sequestration of VCP with polyGA aggregates, altering levels of soluble valosin-containing protein. VCP also functions in autophagy processes, and consistent with this, we observed altered autophagy in cells expressing polyGA. We also observed altered co-localization of polyGA aggregates and p62 in cells depleted of VCP. All together, these data suggest that sequestration of VCP with polyGA aggregates contributes to the loss of VCP function, and consequently to alterations in autophagy processes in C9orf72 expansion disorders.
Summary
Chemokines are key signals in the immune system and play an important role as proinflammatory mediators in the pathology of inflammatory bowel disease and colorectal cancer, making them an ...important target for therapy. Recombinant lactic acid bacteria (LAB) were engineered to bind CC and CXC chemokines by displaying chemokine‐binding proteins evasin‐1, evasin‐3 and evasin‐4 on their surface. Evasin genes were cloned into lactococcal surface display vector and overexpressed in L. lactis NZ9000 and NZ9000ΔhtrA in fusion with secretion signal and surface anchor. Evasin‐displaying bacteria removed from 15% to 90% of 11 different chemokines from the solution as determined with ELISA and Luminex multiplexing assays, whereby L. lactis NZ9000ΔhtrA proved more efficient. Lactobacillus salivarius ATCC 11741 was coated with L. . lactis‐expressed evasin fusion protein, and its ability to bind chemokines was also confirmed. Evasin‐3‐displaying L. lactis removed 76.0% of IL‐1β‐induced CXCL8 from the supernatant of Caco‐2 epithelial cells. It also prevented secretion of CXCL8 from Caco‐2 cells in a time‐dependent manner when added before induction with IL‐1β. Evasin‐displaying LAB have the ability to bind multiple chemokines simultaneously and exert synergistic activity. This innovative treatment approach therefore has the potential for mucosal therapy of inflammatory bowel disease or colorectal cancer.
Recombinant lactic acid bacteria were engineered to bind CC and CXC chemokines by displaying chemokine binding proteins evasins on their surface. Evasin‐displaying bacteria removed from 15% to 90% of 11 different chemokines from the solution and 76.0% of IL‐1β‐induced CXCL8 from the supernatant of Caco‐2 epithelial cells. This innovative treatment approach has the potential for mucosal therapy of inflammatory bowel disease or colorectal cancer.
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