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Abstract ADAM10 and ADAM17 have been shown to contribute to the acquired drug resistance of HER2-positive breast cancer in response to trastuzumab. The majority of ADAM10 and ADAM17 inhibitor development has been focused on the discovery of compounds that bind the active site zinc, however, in recent years, there has been a shift from active site to secondary substrate binding site (exosite) inhibitor discovery in order to identify non-zinc-binding molecules. In the present work a glycosylated, exosite-binding substrate of ADAM10 and ADAM17 was utilized to screen 370,276 compounds from the MLPCN collection. As a result of this uHTS effort, a selective, time-dependent, non-zinc-binding inhibitor of ADAM10 with Ki = 883 nM was discovered. This compound exhibited low cell toxicity and was able to selectively inhibit shedding of known ADAM10 substrates in several cell-based models. We hypothesize that differential glycosylation of these cognate substrates is the source of selectivity of our novel inhibitor. The data indicate that this novel inhibitor can be used as an in vitro and, potentially, in vivo, probe of ADAM10 activity. Additionally, results of the present and prior studies strongly suggest that glycosylated substrate are applicable as screening agents for discovery of selective ADAM probes and therapeutics.

The results of extensive studies that analyzed the existence and meaning of correlations between the economic growth and the financial market development lead us to a more thorough study of these correlations. Therefore, we performed a broad study of the developing countries from around the world (the developing part of each region constructed by the World Bank through its Statistics Bureau). The regions taken into analysis were: Europe and Central Asia, South Asia, East Asia and the Pacific, the Arab world, Latin America & and the Caribbean, the Middle East and North Africa, and Sub-Saharan Africa. For comparison purposes, we have also included in the sample the North American countries, the Euro Area and the European Union as a whole, because these last three areas are the main benchmarks of the financial markets. The results are consistent with those from previous studies on the subject and vary depending on region and financial indicator considered.

Matrix metalloproteinases (MMP) 2 and 9, the gelatinases, have consistently been associated with tumor progression. The development of gelatinase-specific probes will be critical for identifying in vivo gelatinoic activity to understand the molecular role of the gelatinases in tumor development. Recently, a self-assembling homotrimeric triple-helical peptide (THP), incorporating a sequence from type V collagen, with high substrate specificity to the gelatinases has been developed. To determine whether this THP would be suitable for imaging protease activity, 5-carboxyfluorescein (5FAM) was conjugated, resulting in 5FAM3-THP and 5FAM6-THP, which were quenched up to 50%. 5FAM6-THP hydrolysis by MMP-2 and MMP-9 displayed kcat/KM values of 1.5 × 104 and 5.4 × 103 M−1 s−1, respectively. Additionally 5FAM6-THP visualized gelatinase activity in gelatinase positive HT-1080 cells, but not in gelatinase negative MCF-7 cells. Furthermore, the fluorescence in the HT-1080 cells was greatly attenuated by the addition of a MMP-2 and MMP-9 inhibitor, SB-3CT, indicating that the observed fluorescence release was mediated by gelatinase proteolysis and not non-specific proteolysis of the THPs. These results demonstrate that THPs fully substituted with fluorophores maintain their substrate specificity to the gelatinases in human cancer cells and may be useful in in vivo molecular imaging of gelatinase activity.

European Structural Funds have been created with the clear goal to redistribute money among European Union members in such a way as to help poorer regions diminish the gaps between them and the richer ones. The present study assesses this problem for the Structural Operational Programme Environment in Romania. Using spatial econometrics methods, we evaluate two issues on the sample of 42 Romanian counties. The first one is if any clusterization process takes place in what regards contracting funds through ESF for environmental protection projects. The second one is intended to test whether the main principle of the ESF is satisfied – helping the poor in a sustainable development process. Using the quartile maps and the spatial Moran's I autocorrelation coefficient we show different results for the clusterization topic. When considering the budget value of the projects implemented through SOP ENV, both as total and as eligible share, Romanian counties group based on a positive global spatial autocorrelation. This means that counties with approximately the same budget levels are neighbours. Transylvanian counties have gained the most out of the projects on environment. When the number of contracts is assessed, the distribution of the counties proves to be random and no global spatial autocorrelation was found. Moreover, the map analysis shows that, in general, there is a negative relationship between the value of the budget and the number of contracts. The spatial regression analysis shows the violation of the ESF principle. The coefficients of the GDP are positive. This implies a direct relationship between the richness of the county and the projects implemented through the SOP ENV. Thus, instead of being used by the poorer regions, money goes to the richer ones. The higher the GDP, the higher the value of the budget for the environmental protection programmes financed through European funds. The GDP also positively influences the number of contracts scheduled or signed, but the only at a 10% significance level.

The article emphasizes aspects regarding the perceptions of master accountancy graduates concerning the chances to find an adequate job at graduation. These preoccupations are relatively recent in the academic studies. The studentn#8217;s expectations are often unrealistic, which causes problems to them and also to the private firms and organizations. From the empirical section results that the studentsn#8217; evaluations regarding their own opportunities to get a job modify as they advance in the programme. Even more, this evolution is influenced by employment during the studies and by the professional experience.

The teaching performance is influenced firstly by the quality of the educational system. The universities are trying to raise the expectations regarding the methods used for evaluating the teaching quality. In order for the results to be as much as credible the evaluation must defer to the existing theory and to follow the standardized procedures, starting from some basic principles. The most used framework is represented by the 3P model which contains three parts: 1. The input (factors that come under the students and the professor) 2. The learning process (activities focused on learning, teaching methods, practical ability, individual study) 3. The outcome (the output of the learning process).This study presents some results of a questionnaire based on the existing economic theory and applied to a sample of master business students from Romania. The students were asked to give grades from 1 to 5 to several aspects related to the educational process. These aspects took into consideration the following: if the didactic activity was interesting for the students/ the relationship between the objectives of the course and the activity evolved/ the clarity of the presentations/ the degree of students' participation required by the teacher/ the methods and materials used/ the availability of the teacher. The average grade is 4.39, indicating a very high level of satisfaction, taking into account that the grades ranged from 1 to 5. For 50% of the students, the average grade was below 4.53, while for the other half it was above the specified value. But one has to see the value returned for the majority of the students. The mode equals 5, meaning that most of the students involved in the analyzed programme are extremely satisfied with the quality of the educational process. With a significance value Sig. = 0.004 /_ 0.01, we can state that there is correlation between the level of satisfaction expressed by the average grade and the number of classes missed by a student, at a confidence of 99%. The value of the Pearson coefficient is -0.213, which shows the existence of a weak reverse connection.

The article emphasizes aspects regarding the evaluation of the higher education's quality. In certain countries, the questionnaires regarding quality of the activity of HEIs (Higher Education Institutions) are administrated by specialized institutions led by the Ministry of Education or the university associations. The evaluation principles derive from well-known economic and social theories, evolving even evaluation models (see the SERVQUAL model). As a result of the Bologna Treaty (1999), the European Union has the objective to become an international reference concerning the higher education quality and to be more attractive than in the present for students, professors and researchers from other regions of the world. So as to fulfill these objectives ENQA (European Association for Quality Assurance in Higher Education) recommends HEIs to include in their development plans regarding quality aspects five principles described in the article. The practical study refers to the results of a questionnaire applied to the master business students from a Romanian university. In order to assess the level of satisfaction of students in relation to the master programme they are involved in, a questionnaire was applied on a sample of 200 such persons. The responses were then analyzed using multidimensional data analysis methods. Out of these, the present research is based on multiple response analysis. In the questionnaire, students were asked to return their level of satisfaction for different aspects related to the educational process they are involved in. The questions were constructed as five-level Likert items. In this way was insured a connection between answers given at each of the questions assessing the quality of the programme. Only 0.2% of the answers given relate to aspects about which the students were not satisfied at all. These answers represent 3.2% of the number of respondents. 30% of the students were slightly satisfied, returning 57 choices of this type. As one can see, the volume of each group increases with the level of satisfaction. The processing of the questionnaire was made on recent economic theories concerning the quality assurance in the university environment, which are briefly presented in the beginning of the article.

The amyloid-β precursor protein (APP) undergoes proteolysis by β- and γ-secretases to form amyloid-β peptides (Aβ), which is a hallmark of Alzheimer’s disease (AD). Recent findings suggest a possible role of O-glycosylation on APP’s proteolytic processing and subsequent fate for AD-related pathology. We have previously reported that Tyr681-O-glycosylation and the Swedish mutation accelerate cleavage of APP model glycopeptides by β-secretase (amyloidogenic pathway) more than α-secretase (non-amyloidogenic pathway). Therefore, to further our studies, we have synthesized additional native and Swedish-mutated (glyco)peptides with O-GalNAc moiety on Thr663 and/or Ser667 to explore the role of glycosylation on conformation, secretase activity, and aggregation kinetics of Aβ40. Our results show that conformation is strongly dependent on external conditions such as buffer ions and solvent polarity as well as internal modifications of (glyco)peptides such as length, O-glycosylation, and Swedish mutation. Furthermore, the level of β-secretase activity significantly increases for the glycopeptides containing the Swedish mutation compared to their nonglycosylated and native counterparts. Lastly, the glycopeptides impact the kinetics of Aβ40 aggregation by significantly increasing the lag phase and delaying aggregation onset, however, this effect is less pronounced for its Swedish-mutated counterparts. In conclusion, our results confirm that the Swedish mutation and/or O-glycosylation can render APP model glycopeptides more susceptible to cleavage by β-secretase. In addition, this study sheds new light on the possible role of glycosylation and/or glycan density on the rate of Aβ40 aggregation.

Objective: the pourpose of the study was to determine if there are any differences between placenta derived plasmatic levels of messenger RNA in normal and future preeclamptic pregnancies and if these placental transcripts can predict preeclampsia long before clinical onset

After a period of studying a certain subject, students form an opinion about it and begin having certain expectations. These expectations and the degree in which, in the end, they fulfil, contribute to the reputation of the university. Consequently, a continuous evaluation of the quality of the educational process is needed. The present research presents a part of a more complex study made on a sample of master students in Audit and Financial Management in Romania. The goal was to evidence the main factors that affect students' expectations in what regards the grades they will obtain at the end of the semester. For this, a questionnaire of 20 questions was applied to 250 such students. After factor reduction procedures were applied, six most significant variables were kept in the analysis: the proportion of knowledge acquired, the perceived level of utility of the discipline in the professional career of the student, the proportion in which the subject could contribute to getting employed in the field it belongs to, the evaluation method and two variables evaluating through grades the didactic performance during the course and the overall performance of the tenure professor. The influence of these variables upon the grade expected by the student was assessed with the help of the OLS regression, both in the simple and multiple forms. Out of the six hypotheses formulated, only one proved to be false based on the simple regression analysis. When individually assessed, the evaluation method announced by the teacher at the beginning of the semester turned out to have no statistically significant influence upon students' expectations. For the rest of the variables, results were according to the assumptions made, i.e. all determine in a significant positive manner the students' opinion about the grade they will get. We have also constructed the multiple regression models. When putting all variables together, the significance changes. The level of difficulty of the evaluation method becomes significant, while from the rest of the variables only the proportion of knowledge acquirement still holds. The final conclusion is as expected: the higher the proportion of acquirement, the higher the grade expected at the end of the semester, while the more difficult the evaluation method, the lower the grade.

The study shows the reasons taken into account by the students in choosing an academic preparation. Free courses and employment opportunities during or after graduation are the most important advantages students expect to receive from an educational program that aims to follow. The empirical section results emphasize the efficiency and performance of the master program financed by the European Social Fund. \\\\r\\\\n

Rice blast, caused by Pyricularia oryzae , is one of the main rice diseases worldwide. The pyramiding of blast-resistance ( Pi ) genes, coupled to Marker-Assisted BackCrossing (MABC), provides broad-spectrum and potentially durable resistance while limiting the donor genome in the background of an elite cultivar. In this work, MABC coupled to foreground and background selections based on KASP marker assays has been applied to introgress four Pi genes ( Piz, Pib, Pita , and Pik ) in a renowned japonica Italian rice variety, highly susceptible to blast. Molecular analyses on the backcross (BC) lines highlighted the presence of an additional blast-resistance gene, the Pita -linked Pita2 / Ptr gene, therefore increasing the number of blast-resistance introgressed genes to five. The recurrent genome was recovered up to 95.65%. Several lines carrying four (including Pita2 ) Pi genes with high recovery percentage levels were also obtained. Phenotypic evaluations confirmed the effectiveness of the pyramided lines against multivirulent strains, which also had broad patterns of resistance in comparison to those expected based on the pyramided Pi genes. The developed blast-resistant japonica lines represent useful donors of multiple blast-resistance genes for future rice-breeding programs related to the japonica group.

Background: Rice is one of the most salt sensitive crops at seedling, early vegetative and reproductive stages. Varieties with salinity tolerance at seedling stage promote an efficient growth at early stages in salt affected soils, leading to healthy vegetative growth that protects crop yield. Saltol major QTL confers capacity to young rice plants growing under salt condition by maintaining a low Na + /K + molar ratio in the shoots., Results: Marker-assisted backcross (MABC) procedure was adopted to transfer Saltol locus conferring salt tolerance at seedling stage from donor indica IR64-Saltol to two temperate japonica varieties, Vialone Nano and Onice. Forward and background selections were accomplished using polymorphic KASP markers and a final evaluation of genetic background recovery of the selected lines was conducted using 15,580 SNP markers obtained from Genotyping by Sequencing. Three MABC generations followed by two selfing, allowed the identification of introgression lines achieving a recovery of the recurrent parent (RP) genome up to 100% (based on KASP markers) or 98.97% (based on GBS). Lines with highest RP genome recovery (RPGR) were evaluated for agronomical-phenological traits in field under non-salinized conditions. VN1, VN4, O1 lines were selected considering the agronomic evaluations and the RPGR% results as the most interesting for commercial exploitation. A physiological characterization was conducted by evaluating salt tolerance under hydroponic conditions. The selected lines showed lower standard evaluation system (SES) scores: 62% of VN4, and 57% of O1 plants reaching SES 3 or SES 5 respectively, while only 40% of Vialone Nano and 25% of Onice plants recorded scores from 3 to 5, respectively. VN1, VN4 and O1 showed a reduced electrolyte leakage values, and limited negative effects on relative water content and shoot/root fresh weight ratio., Conclusion: The Saltol locus was successfully transferred to two elite varieties by MABC in a time frame of three years. The application of background selection until BC 3 F 3 allowed the selection of lines with a RPGR up to 98.97%. Physiological evaluations for the selected lines indicate an improved salinity tolerance at seedling stage. The results supported the effectiveness of the Saltol locus in temperate japonica and of the MABC procedure for recovering of the RP favorable traits., (© 2023. The Author(s).)

Rice is unique among cereals for its ability to germinate not only when submerged but also under anoxic conditions. Rice germination under submergence or anoxia is characterized by a longer coleoptile and delay in radicle emergence. A panel of temperate and tropical japonica rice accessions showing a large variability in coleoptile length was used to investigate genetic factors involved in this developmental process. The ability of the Khao Hlan On rice landrace to vigorously germinate when submerged has been previously associated with the presence of the trehalose 6 phosphate phosphatase 7 (TPP7) gene. In this study, we found that, in the presence of TPP7, polymorphisms and transcriptional variations of the gene in coleoptile tissue were not related to differences in the final coleoptile length under submergence. In order to find new chromosomal regions associated with the different ability of rice to elongate the coleoptile under submergence, we used genome-wide association study analysis on a panel of 273 japonica rice accessions. We discovered 11 significant marker-trait associations and identified candidate genes potentially involved in coleoptile length. Candidate gene expression analyses indicated that japonica rice genotypes possess complex genetic elements that control final coleoptile length under low oxygen., (© 2019 John Wiley & Sons Ltd.)

The domestication of wild emmer wheat led to the selection of modern durum wheat, grown mainly for pasta production. We describe the 10.45 gigabase (Gb) assembly of the genome of durum wheat cultivar Svevo. The assembly enabled genome-wide genetic diversity analyses revealing the changes imposed by thousands of years of empirical selection and breeding. Regions exhibiting strong signatures of genetic divergence associated with domestication and breeding were widespread in the genome with several major diversity losses in the pericentromeric regions. A locus on chromosome 5B carries a gene encoding a metal transporter (TdHMA3-B1) with a non-functional variant causing high accumulation of cadmium in grain. The high-cadmium allele, widespread among durum cultivars but undetected in wild emmer accessions, increased in frequency from domesticated emmer to modern durum wheat. The rapid cloning of TdHMA3-B1 rescues a wild beneficial allele and demonstrates the practical use of the Svevo genome for wheat improvement.

Background: Leaf pigment content is an important trait involved in environmental interactions. In order to determine its impact on drought tolerance in wheat, we characterized a pale-green durum wheat mutant (Triticum turgidum L. var. durum) under contrasting water availability conditions., Results: The pale-green mutant was investigated by comparing pigment content and gene/protein expression profiles to wild-type plants at anthesis. Under well-watered (control) conditions the mutant had lower levels of chlorophylls and carotenoids, but higher levels of xanthophyll de-epoxidation compared to wild-type. Transcriptomic analysis under control conditions showed that defense genes (encoding e.g. pathogenesis-related proteins, peroxidases and chitinases) were upregulated in the mutant, suggesting the presence of mild oxidative stress that was compensated without altering the net rate of photosynthesis. Transcriptomic analysis under terminal water stress conditions, revealed the modulation of antioxidant enzymes, photosystem components, and enzymes representing carbohydrate metabolism and the tricarboxylic acid cycle, indicating that the mutant was exposed to greater oxidative stress than the wild-type plants, but had a limited capacity to respond. We also compared the two genotypes under irrigated and rain-fed field conditions over three years, finding that the greater oxidative stress and corresponding molecular changes in the pale-green mutant were associated to a yield reduction., Conclusions: This study provides insight on the effect of pigment content in the molecular response to drought. Identified genes differentially expressed under terminal water stress may be valuable for further studies addressing drought resistance in wheat.

Background: Durum wheat often faces water scarcity and high temperatures, two events that usually occur simultaneously in the fields. Here we report on the stress responsive strategy of two durum wheat cultivars, characterized by different water use efficiency, subjected to drought, heat and a combination of both stresses., Results: The cv Ofanto (lower water use efficiency) activated a large set of well-known drought-related genes after drought treatment, while Cappelli (higher water use efficiency) showed the constitutive expression of several genes induced by drought in Ofanto and a modulation of a limited number of genes in response to stress. At molecular level the two cvs differed for the activation of molecular messengers, genes involved in the regulation of chromatin condensation, nuclear speckles and stomatal closure. Noteworthy, the heat response in Cappelli involved also the up-regulation of genes belonging to fatty acid β-oxidation pathway, glyoxylate cycle and senescence, suggesting an early activation of senescence in this cv. A gene of unknown function having the greatest expression difference between the two cultivars was selected and used for expression QTL analysis, the corresponding QTL was mapped on chromosome 6B., Conclusion: Ofanto and Cappelli are characterized by two opposite stress-responsive strategies. In Ofanto the combination of drought and heat stress led to an increased number of modulated genes, exceeding the simple cumulative effects of the two single stresses, whereas in Cappelli the same treatment triggered a number of differentially expressed genes lower than those altered in response to heat stress alone. This work provides clear evidences that the genetic system based on Cappelli and Ofanto represents an ideal tool for the genetic dissection of the molecular response to drought and other abiotic stresses.

WRKY proteins constitute a large family of plant specific transcription factors implicated in many different processes. Here we describe Hv-WRKY38, a barley gene coding for a WRKY protein, whose expression is involved in cold and drought stress response. Hv-WRKY38 was early and transiently expressed during exposure to low non-freezing temperature, in ABA-independent manner. Furthermore, it showed a continuous induction during dehydration and freezing treatments. A WRKY38:YFP fusion protein was found to localise into the nucleus upon introduction into epidermal onion cells. Bacterially expressed Hv-WRKY38 was able to bind in vitro to the W-box element (T)TGAC(C/T) also recognisable by other WRKY proteins. Hv-WRKY38 genomic DNA was sequenced and mapped onto the centromeric region of the barley chromosome 6H. Arabidopsis and rice sequences homologous to Hv-WRKY38 were also identified. Our results indicate that Hv-WRKY38 transcription factor may play a regulatory role in abiotic stress response.

The accumulation of specific cold-regulated (COR) proteins is a component of the hardening process and different amount of COR proteins has been related to different degrees of cold tolerance. A number of different mechanisms controls the accumulation of the COR proteins in the plant cells. In this work we describe the mechanisms controlling the accumulation of the COR protein TMC-AP3, a putative chloroplastic amino acid selective channel protein [1] in barley, durum, wheat, emmer and bread wheat. Winter barley and, to less extent, winter bread wheat showed a higher cor tmc-ap3 expression at low temperature than the spring one while no significant differences were detected between the emmer and the durum. wheat genotypes. After 2 days of de-hardening the transcript level dropped down in the same way in all tested genotypes, nevertheless the decrease in protein content was genotype dependent. In all frost resistant genotypes the amount of COR TMC-AP3 after 9 days of de-hardening was higher compared with that of susceptible ones. These findings suggest that resistant and susceptible genotypes have different protein degradation rate and/or mRNA translational efficiency. Differences in the protein degradation rate were not dependent from the amino acidic sequence of the protein, being extremely similar in all tested genotypes. A genetic study based on Chinese spring/Cheyenne chromosome substitution lines showed that the turnover of TMC-AP3 is a polygenic trait controlled by a number of loci being the most important located on chromosomes 1B, 2B, 2D and 4D.