Luteinising hormone-releasing hormone analogue reverses the cell adhesion profile of EGFR overexpressing DU-145 human prostate carcinoma subline

Cetrorelix, a luteinising hormone-releasing hormone (LHRH) analogue, has been shown to limit growth of the human androgen-independent prostate cell line DU-145, although other inhibitory actions may also be affected. Both growth and invasion of DU-145 cells are linked to autocrine epidermal growth factor receptor (EGFR) signalling. Invasiveness requires not only cells to migrate to conduits, but also reduced adhesiveness between tumour cells to enable separation from the tumour mass. Thus, we investigated whether Cetrorelix alters the DU-145 cell–cell adhesion and if this occurs via altered EGFR signalling. Pharmacologic levels of Cetrorelix limited the invasiveness of a highly invasive DU-145 subline overexpressing full-length EGFR (DU-145 WT). Extended exposure of the cells to Cetrorelix resulted in increased levels of the cell–cell adhesion complex molecules E-cadherin, α- and β-catenin, and p120. Puromycin blocked the increases in E-cadherin and β-catenin levels, suggesting that de novo protein synthesis is required. The Cetrorelix effect appears to occur via transmodulation of EGFR by a protein kinase C (PKC)-dependent mechanism, as there were no changes in DU-145 cells expressing EGFR engineered to negate the PKC transattenuation site (DU-145 A654); downregulation of EGFR signalling produced a similar upregulation in adhesion complex proteins, further suggesting a role for autocrine signalling. Cetrorelix increased the cell–cell adhesiveness of DU-145 WT cells to an extent similar to that seen when autocrine EGFR signalling is blocked; as expected, DU-145 A654 cell–cell adhesion also was unaffected by Cetrorelix. The increased adhesiveness is expected as the adhesion complex molecules moved to the cells' periphery. These data offer direct insight into the possible crosstalk pathways between the LHRH and EGFR receptor signalling. The ability of Cetrorelix to downregulate EGFR signalling and subsequently reverse the antiadhesiveness found in metastatic prostate cancer highlights a novel potential target for therapeutic strategies

Structural and functional analysis of the C-terminal STAS domain of the Arabidopsis thaliana sulfate transporter SULTR1.2

The C-terminal region of sulfate transporters from plants and animals belonging to the SLC26 family members shares a weak but significant similarity with the Bacillus sp. anti-anti-sigma protein SpoIIAA, thus defining the STAS domain (Sulfate Transporter and Anti Sigma antagonist). The present study is a structure/function analysis of the STAS domain of SULTR1.2, an Arabidopsis thaliana sulfate transporter. A 3-D model of the SULTR1.2 STAS domain was built, which indicated that it shares the SpoIIAA folds. Moreover, the phosphorylation site, which is necessary for SpoIIAA activity, is conserved in the SULTR1.2 STAS domain. The model was used to direct mutagenesis studies using a yeast mutant defective for sulfate transport. Truncation of the whole SULTR1.2 STAS domain resulted in the loss of sulfate transport function. Analyses of small deletions and mutations showed that the C-terminal tail of the SULTR1.2 STAS domain, and particularly two cysteine residues, plays an important role in sulfate transport by SULTR1.2. All the substitutions made at the putative phosphorylation site Thr587 led to a complete loss of the sulfate transport function of SULTR1.2. The reduction or suppression of sulfate transport of the SULTR1.2 mutants in yeast was not due to an incorrect targeting to the plasma membrane. Both our 3D modeling and mutational analyses strengthen the hypothesis that the SULTR1.2 STAS domain is involved in protein-protein interactions that could control sulfate transport

An ABC Transporter Mutation Alters Root Exudation of Phytochemicals That Provoke an Overhaul of Natural Soil Microbiota1[C][W][OA]

Root exudates influence the surrounding soil microbial community, and recent evidence demonstrates the involvement of ATP-binding cassette (ABC) transporters in root secretion of phytochemicals. In this study, we examined effects of seven Arabidopsis (Arabidopsis thaliana) ABC transporter mutants on the microbial community in native soils. After two generations, only the Arabidopsis abcg30 (Atpdr2) mutant had significantly altered both the fungal and bacterial communities compared with the wild type using automated ribosomal intergenic spacer analysis. Similarly, root exudate profiles differed between the mutants; however, the largest variance from the wild type (Columbia-0) was observed in abcg30, which showed increased phenolics and decreased sugars. In support of this biochemical observation, whole-genome expression analyses of abcg30 roots revealed that some genes involved in biosynthesis and transport of secondary metabolites were up-regulated, while some sugar transporters were down-regulated compared with genome expression in wild-type roots. Microbial taxa associated with Columbia-0 and abcg30 cultured soils determined by pyrosequencing revealed that exudates from abcg30 cultivated a microbial community with a relatively greater abundance of potentially beneficial bacteria (i.e. plant-growth-promoting rhizobacteria and nitrogen fixers) and were specifically enriched in bacteria involved in heavy metal remediation. In summary, we report how a single gene mutation from a functional plant mutant influences the surrounding community of soil organisms, showing that genes are not only important for intrinsic plant physiology but also for the interactions with the surrounding community of organisms as well

Cyanide Biodegradation by a Native Bacterial Consortium and Its Potential for Goldmine Tailing Biotreatment

A native cyanide-degrading bacterial consortium was isolated from goldmine tailing sediments. Mine tailings are toxic effluents due to their metal–cyanide complexes. The bacterial consortium was able to degrade an initial sodium cyanide concentration ranging from 5 to 120 mg L−1 in alkaline synthetic wastewater (pH > 9.2), for a maximum of 15 days. The free cyanide biodegradation efficiency was 98% for the highest initial free cyanide concentration tested and followed a first-order kinetic profile, with an estimated kinetic rate constant of 0.12 ± 0.011 d−1. The cyanide-degrading consortium was streaked with serial dilutions on a specific medium (R2A). 16S rRNA gene sequencing and mass spectrometry proteomic fingerprinting of the isolates showed that the bacterial strains belonged to Microbacterium paraoxydans, Brevibacterium casei, Brevundimonas vesicularis, Bacillus cereus and Cellulosimicrobium sp. The first four genera had previously been identified as cyanide-degrading bacteria. Microbacterium and Brevibacterium had previously been found in alkaline conditions, showing resistance to heavy metals. As for Cellulosimicrobium, to our knowledge, this is the first study to implicate it directly or indirectly in cyanide biodegradation. In this research, these genera were identified as functional bacteria for cyanide degradation, and they might be suitable for mine tailing biotechnological tertiary treatment

Nivel tecnológico de invernadero y riesgo para la salud de los jornaleros

Abstract Introduction: The greenhouses are classified by under high levels or as technologies to be adapted to create optimal conditions of radiation, temperature, humidity and carbon dioxide that improve agricultural production and the development of organisms harmful that fight applying pesticides, exposing laborers to vapors and waste. Method: The cytotoxic damage was determined in samples of oral epithelial cells exfoliation of day laborers through the micronucleus assay, 40 laborers exposed to pesticides, 20 worked in greenhouses low-tech and high-tech 20. Results: The low-tech results showed higher frequency for all nuclear abnormalities compared to the control group, especially of condensed chromatin, picnosis, karyolysis and micronuclei. In calculating the risk, the frequency of micronuclei was measured at odds ratio (OR) for laborers in high-tech greenhouses OR = 17.06 (95 % CI 3.4-83.7) and low-tech OR = 50.66 (% CI 7.5 -341.7). Conclusion: The results indicate that the laborers who work in low-tech greenhouses show a higher frequency of cells with nuclear abnormalities, due to the effect of working conditions and exposure to pesticides.Resumen Introducción: Los invernaderos se clasifican por niveles alto, medio o bajo según las tecnologías que se le adapten, con la que se crean condiciones óptimas de radiación, temperatura, humedad y dióxido de carbono que permiten mejorar la producción agrícola, así como el desarrollo de organismos perjudiciales que se combaten aplicando plaguicidas, exponiendo a los jornaleros a sus vapores y residuos. Método: Se determinó el daño genotóxico en muestras de células epiteliales de exfoliación bucal de los jornaleros a través del ensayo de micronúcleos, de 40 jornaleros expuestos a plaguicidas, 20 laboraron en invernaderos de baja tecnología y 20 en invernaderis de tecnología alta. Resultados: Los de baja tecnología mostraron resultados más altos de frecuencias para todas las anormalidades nucleares comparando con el grupo control, especialmente de cromatina condensada, picnosis, cariolisis y micronúcleos. Al calcular el riesgo, se midió las frecuencias de micronúcleos en razón de probabilidades (OR), para los jornaleros de invernaderos de alta tecnología el OR = 17.06 (95 % IC 3.4-83.7) y de baja tecnología OR = 50.66 (% IC 7.5-341.7). Conclusión: Los resultados indican que los jornaleros que laboran en invernaderos de baja tecnología muestran una mayor frecuencia de células con anormalidades nucleares, debido al efecto de condiciones laborales y exposición a los plaguicidas

Characteristics of innovation policy mixes in multi-level government system: the case of the Baltic Sea Region countries

Elektroniskā versija nesatur pielikumusPromocijas darbā pētīts, kā notiek inovācijas rīcībpolitikas paralēla plānošana un ieviešana dažādos pārvaldes līmeņos – vietējā, nacionālā un pārnacionālā. Analizētas divas gadījumu grupas – Baltijas valstis un Somija, Zviedrija un Dānija – kas atšķiras pēc inovācijas rīcībpolitikas īstenošanā aktīvi iesaistīto pārvaldes līmeņu skaita. Darba teorētiskais ietvars ir daudzlīmeņu pārvaldības un rīcībpolitiku kombināciju koncepti, kas izmantoti kvalitatīvo datu analīzes kategoriju izstrādei. Gadījumi analizēti un salīdzināti, izmantojot politikas plānošanas dokumentus un veicot padziļinātās intervijas ar rīcībpolitikas veidotājiem katrā pārvaldes līmenī. Hipotēze, kurā tika pieņemts, ka daudzlīmeņu inovācijas rīcībpolitikas kombinācijas raksturo nesaskaņotība, netika pierādīta, taču starp pārvaldes līmeņiem nevar novērot arī pozitīvu sinerģiju.Dissertation explores how innovation policy is planned and implemented at different levels of government - local, national and transnational. Two groups of cases are analysed - the Baltic States and Denmark, Sweden and Finland. The groups of cases differ according to the number of government levels involved in the innovation policy making. The theoretical frameworks of the thesis are the concepts of policy mix and multi-level governance. Both are used to develop the categories for qualitative data analysis. The cases are analysed and compared using the policy planning documents and conducting in-depth interviews with policy makers at each level of government. The hypothesis, which assumed that multi-level innovation policy mixes are incoherent, was not proved, however, interaction between government levels was not identified either