Effect of gibberellic acid and the biostimulant stimulate® on the initial growth of tamarind

Plant growth regulators and biostimulants have been used as an agronomic technique to optimize the production of seedlings in various crops. This study aimed to evaluate the influence of gibberellic acid and the biostimulant Stimulate® on the initial growth of tamarind (Tamarindus indica L.). The experiments were conducted in a nursery with 50% shading, in a randomized block design with five replications and five plants per plot. Thirty eight days after sowing, the leaves were sprayed seven times a day with 0.0 (control), 0.8, 1.6, 2.4 and 3.2 mL of gibberellic acid L-1 aqueous solution and with 0.0 (control), 6.0,12.0, 18.0, and 24.0 mL Stimulate® L-1 aqueous solution. Stem diameter (SD), plant height (PH), longest root length (LRL), shoot dry mass (SDM), root dry mass (RDM) and RDM:SDM ratio were evaluated ninety days after sowing. Variance and regression analysis showed that GA3 at 4% promoted plant growth (height), but had no significant effect on stem diameter, longest root length, shoot and root dry mass and the RDM:SDM ratio. On the other hand, all concentrations of Stimulate® significantly increased plant height and shoot and root dry mass of tamarind seedlings.

Homozygosity mapping reveals novel and known mutations in Pakistani families with inherited retinal dystrophies.

Inherited retinal dystrophies are phenotypically and genetically heterogeneous. This extensive heterogeneity poses a challenge when performing molecular diagnosis of patients, especially in developing countries. In this study, we applied homozygosity mapping as a tool to reduce the complexity given by genetic heterogeneity and identify disease-causing variants in consanguineous Pakistani pedigrees. DNA samples from eight families with autosomal recessive retinal dystrophies were subjected to genome wide homozygosity mapping (seven by SNP arrays and one by STR markers) and genes comprised within the detected homozygous regions were analyzed by Sanger sequencing. All families displayed consistent autozygous genomic regions. Sequence analysis of candidate genes identified four previously-reported mutations in CNGB3, CNGA3, RHO, and PDE6A, as well as three novel mutations: c.2656C > T (p.L886F) in RPGRIP1, c.991G > C (p.G331R) in CNGA3, and c.413-1G > A (IVS6-1G > A) in CNGB1. This latter mutation impacted pre-mRNA splicing of CNGB1 by creating a -1 frameshift leading to a premature termination codon. In addition to better delineating the genetic landscape of inherited retinal dystrophies in Pakistan, our data confirm that combining homozygosity mapping and candidate gene sequencing is a powerful approach for mutation identification in populations where consanguineous unions are common.

Gene expression following induction of regeneration in Drosophila wing imaginal discs

Background: Regeneration is the ability of an organism to rebuild a body part that has been damaged or amputated, and can be studied at the molecular level using model organisms. Drosophila imaginal discs, which are the larval primordia of adult cuticular structures, are capable of undergoing regenerative growth after transplantation and in vivo culture into the adult abdomen. Results: Using expression profile analyses, we studied the regenerative behaviour of wing discs at 0, 24 and 72 hours after fragmentation and implantation into adult females. Based on expression level, we generated a catalogue of genes with putative role in wing disc regeneration, identifying four classes: 1) genes with differential expression within the first 24 hours; 2) genes with differential expression between 24 and 72 hours; 3) genes that changed significantly in expression levels between the two time periods; 4) genes with a sustained increase or decrease in their expression levels throughout regeneration. Among these genes, we identified members of the JNK and Notch signalling pathways and chromatin regulators. Through computational analysis, we recognized putative binding sites for transcription factors downstream of these pathways that are conserved in multiple Drosophilids, indicating a potential relationship between members of the different gene classes. Experimental data from genetic mutants provide evidence of a requirement of selected genes in wing disc regeneration. Conclusions: We have been able to distinguish various classes of genes involved in early and late steps of the regeneration process. Our data suggests the integration of signalling pathways in the promoters of regulated genes.

Long noncoding RNAs in cardiac development and ageing.

A large part of the mammalian genome is transcribed into noncoding RNAs. Long noncoding RNAs (lncRNAs) have emerged as critical epigenetic regulators of gene expression. Distinct molecular mechanisms allow lncRNAs either to activate or to repress gene expression, thereby participating in the regulation of cellular and tissue function. LncRNAs, therefore, have important roles in healthy and diseased hearts, and might be targets for therapeutic intervention. In this Review, we summarize the current knowledge of the roles of lncRNAs in cardiac development and ageing. After describing the definition and classification of lncRNAs, we present an overview of the mechanisms by which lncRNAs regulate gene expression. We discuss the multiple roles of lncRNAs in the heart, and focus on the regulation of embryonic stem cell differentiation, cardiac cell fate and development, and cardiac ageing. We emphasize the importance of chromatin remodelling in this regulation. Finally, we discuss the therapeutic and biomarker potential of lncRNAs.

Gene expression following induction of regeneration in Drosophila wing imaginal discs

Background: Regeneration is the ability of an organism to rebuild a body part that has been damaged or amputated, and can be studied at the molecular level using model organisms. Drosophila imaginal discs, which are the larval primordia of adult cuticular structures, are capable of undergoing regenerative growth after transplantation and in vivo culture into the adult abdomen. Results: Using expression profile analyses, we studied the regenerative behaviour of wing discs at 0, 24 and 72 hours after fragmentation and implantation into adult females. Based on expression level, we generated a catalogue of genes with putative role in wing disc regeneration, identifying four classes: 1) genes with differential expression within the first 24 hours; 2) genes with differential expression between 24 and 72 hours; 3) genes that changed significantly in expression levels between the two time periods; 4) genes with a sustained increase or decrease in their expression levels throughout regeneration. Among these genes, we identified members of the JNK and Notch signalling pathways and chromatin regulators. Through computational analysis, we recognized putative binding sites for transcription factors downstream of these pathways that are conserved in multiple Drosophilids, indicating a potential relationship between members of the different gene classes. Experimental data from genetic mutants provide evidence of a requirement of selected genes in wing disc regeneration. Conclusions: We have been able to distinguish various classes of genes involved in early and late steps of the regeneration process. Our data suggests the integration of signalling pathways in the promoters of regulated genes.

Blueberry production in Chile: current status and future developments

Chile has become a major actor in the blueberry industry as the most important supplier of off-season fresh fruit for the northern hemisphere. Blueberry exports passed from US$ 30 million (around 4,000 tons) in 2000 to US$ 380 million (94,000 tons) in 2011. The characteristics of the major blueberry growing regions (North, Central, South-central and South) are presented in terms of acreage, varieties, management practices, extension of the harvest season, and soil and climatic conditions. Most fruit is from highbush varieties, picked by hand and exported fresh by boat to United States. Largest proportion of fruit is exported from mid December to late January, which coincides with lowest prices. The south-central region (latitudes 34º50' to 38º15' S) was in 2007 the most important one with 5,075 ha (51.1% of area planted). Among the challenges for the Chilean blueberry industry in the near future are: 1. Lower profitability due to lower rates of currency exchange and higher costs, 2 - Greater scarcity and higher cost of labor, 3.- Need for higher productivity and sustainable production practices, 4- Fruit of high and consistent quality, and 5.- Greater investment in research. As a case study the article presents three approaches that can help identify areas with low availability of labor and improve its efficiency. The article shows the use of geomatic tools to establish labor availability, application of growth regulators to reduce crop load, increase fruit size and improve harvest efficiency, and the use of shakers to harvest fresh fruit for long distance markets. More research is needed to improve yields, reduce costs and give greater economical and ecological sustainability to the Chilean blueberry industry.

Metal dealing at the origin of the Chordata phylum: The metallothionein system and metal overload response in Amphioxus.

Non-vertebrate chordates, specifically amphioxus, are considered of the utmost interest for gaining insight into the evolutionary trends, i.e. differentiation and specialization, of gene/protein systems. In this work, MTs (metallothioneins), the most important metal binding proteins, are characterized for the first time in the cephalochordate subphylum at both gene and protein level, together with the main features defining the amphioxus response to cadmium and copper overload. Two MT genes (BfMT1 and BfMT2) have been identified in a contiguous region of the genome, as well as several ARE (antioxidant response element) and MRE (metal response element) located upstream the transcribed region. Their corresponding cDNAs exhibit identical sequence in the two lancelet species (B. floridae and B. lanceolatum), BfMT2 cDNA resulting from an alternative splicing event. BfMT1 is a polyvalent metal binding peptide that coordinates any of the studied metal ions (Zn, Cd or Cu) rendering complexes stable enough to last in physiological environments, which is fully concordant with the constitutive expression of its gene, and therefore, with a metal homeostasis housekeeping role. On the contrary, BfMT2 exhibits a clear ability to coordinate Cd(II) ions, while it is absolutely unable to fold into stable Cu (I) complexes, even as mixed species. This identifies it as an essential detoxification agent, which is consequently only induced in emergency situations. The cephalochordate MTs are not directly related to vertebrate MTs, neither by gene structure, protein similarity nor metal-binding behavior of the encoded peptides. The closest relative is the echinoderm MT, which confirm proposed phylogenetic relationships between these two groups. The current findings support the existence in most organisms of two types of MTs as for their metal binding preferences, devoted to different biological functions: multivalent MTs for housekeeping roles, and specialized MTs that evolve either as Cd-thioneins or Cu-thioneins, according to the ecophysiological needs of each kind of organisms.

Molecular profiling of CD8 T cells in autochthonous melanoma identifies Maf as driver of exhaustion.

T cells infiltrating neoplasms express surface molecules typical of chronically virus-stimulated T cells, often termed "exhausted" T cells. We compared the transcriptome of "exhausted" CD8 T cells infiltrating autochthonous melanomas to those of naïve and acutely stimulated CD8 T cells. Despite strong similarities between transcriptional signatures of tumor- and virus-induced exhausted CD8 T cells, notable differences appeared. Among transcriptional regulators, Nr4a2 and Maf were highly overexpressed in tumor-exhausted T cells and significantly upregulated in CD8 T cells from human melanoma metastases. Transduction of murine tumor-specific CD8 T cells to express Maf partially reproduced the transcriptional program associated with tumor-induced exhaustion. Upon adoptive transfer, the transduced cells showed normal homeostasis but failed to accumulate in tumor-bearing hosts and developed defective anti-tumor effector responses. We further identified TGFβ and IL-6 as main inducers of Maf expression in CD8 T cells and showed that Maf-deleted tumor-specific CD8 T cells were much more potent to restrain tumor growth in vivo. Therefore, the melanoma microenvironment contributes to skewing of CD8 T cell differentiation programs, in part by TGFβ/IL-6-mediated induction of Maf.

Pankkien vakavaraisuussäännöstön uudistus

Tutkielman tavoitteena on kuvata pankkien vakavaraisuusuudistuksen eri osa-alueita. Tarkempi analyysi rajautuu uudistuksen tuomiin muutoksiin luotto- ja operatiivisen riskin pääomavaateissa. Tutkielman empiirisen osuuden tavoitteena on perehtyä vakavaraisuussäännöstön uudistusten vaikutuksiin Nordeassa. Tutkimusmetodologiaksi on valittu normatiivinen tutkimusote. Lisäksi tutkielma sisältää deskriptiivisiä ja positivistisia osia. Lähdeaineisto koostuu Baselin pankkivalvontakomitean ja Suomen Pankin julkaisemista tutkimuksista ja dokumenteista sekä alan julkaisuissa ilmestyneistä artikkeleista. Pankkien vakavaraisuussäännöstöuudistuksen tavoitteena on lisätä rahoitusmarkkinoiden vakautta. Sääntelyn kautta pyritään turvaamaan pankkien varojen riittävyys suhteessa niiden riskien ottoon. Vakavaraisuussäännöstön uudistus muodostuu kolmesta pilarista: (1) minimipääomavaatimuksista, (2) pankkivalvonnan vahvistamisesta ja (3) markkinakurin hyödyntämisestä luottolaitosten toiminnan julkistamisvaatimuksia lisäämällä. Pankkivalvonnan harmonisoinnista vallitsee kansainvälinen yhteisymmärrys, mutta ennen kuin Basel II voi astua voimaan on useita ongelmia ratkaisematta. Baselin vakavaraisuuskehikko ei ole ainut lähitulevaisuudessa pankkitoimialaa koetteleva uudistus. Kansainväliset tilinpäätösstandardit; International Accounting Standards ja erityisesti IAS 39 sekä International Financial Reporting Standards, lyhyemmin IFRS tulevat muuttamaan merkittävästi pankkien tilinpäätöskäyttäytymistä. Epäselvää on vielä kuitenkin tukevatko uudistukset toisiaan ja missä määrin pankkien tulosvolatiliteetin odotetaan kasvavan. Tutkielmassa pohditaan vakavaraisuussäännöstön uudistuksen hyötyjä kansainvälisen kilpailuneutraliteetin osalta, sillä Yhdysvalloissa uudistus koskee vain suurimpia pankkeja. Tutkielmassa paneudutaan lisäksi uudistuksen mahdolliseen talouden syklejä voimistavaan vaikutukseen ja tarkastellaan parannusehdotuksia prosyklisyyden hillitsemiseksi. Yksi vakavaraisuusuudistuksen tärkeimmistä tehtävistä on luoda pankeille kannustin kehittää omia riskienhallinta malleja. Kannustin ongelma on pyritty ratkaisemaan vapaampien sisäisten mallien menetelmien avulla. Ongelmaa ei ole pystytty kuitenkaan ratkaisemaan aivan täysin, sillä luottoriskien osalta pankkien lainaportfolioiden rakenne määrittää sen, hyötyvätkö pankit siirtymisestä sisäisten mallien menetelmän käyttöön. Tutkielma sisältää myös Nordean arvion vakavaraisuusuudistuksen vaikutuksista pankkitoimialaan.

Sox4 participates in the modulation of Schwann cell myelination.

In order to identify new regulators of Schwann cell myelination potentially playing a role in peripheral nervous system (PNS) pathologies, we analysed gene expression profiling data from three mouse models of demyelinating neuropathies and from the developing PNS. This analysis revealed that Sox4, which encodes a member of the Sry-related high-mobility group box protein family, was consistently upregulated in all three analysed models of neuropathy. Moreover, Sox4 showed a peak in its expression during development that corresponded with the onset of myelination. To gain further insights into the role of Sox4 in PNS development, we generated a transgenic mouse that specifically overexpresses Sox4 in Schwann cells. Sox4 overexpression led to a temporary delay in PNS myelination without affecting axonal sorting. Importantly, we observed that, whereas Sox4 mRNA could be efficiently overexpressed, Sox4 protein expression in Schwann cells was strictly regulated. Finally, our data showed that enforced expression of Sox4 in the mouse model for Charcot-Marie-Tooth 4C aggravated its neuropathic phenotype. Together, these observations reveal that Sox4 contributes to the regulation of Schwann cell myelination, and also indicates its involvement in the pathophysiology of peripheral neuropathies.

JNK Inhibition Reduced Retinal Ganglion Cell Death after Ischemia/Reperfusion In Vivo and after Hypoxia In Vitro.

Mitogen-activated protein kinases (MAPKs) are key regulators that have been linked to cell survival and death. Among the main classes of MAPKs, c-jun N-terminal kinase (JNK) has been shown to mediate cell stress responses associated with apoptosis. In Vitro, hypoxia induced a significant increase in 661W cell death that paralleled increased activity of JNK and c-jun. 661W cells cultured in presence of the inhibitor of JNK (D-JNKi) were less sensitive to hypoxia-induced cell death. In vivo, elevation in intraocular pressure (IOP) in the rat promoted cell death that correlated with modulation of JNK activation. In vivo inhibition of JNK activation with D-JNKi resulted in a significant and sustained decrease in apoptosis in the ganglion cell layer, the inner nuclear layer and the photoreceptor layer. These results highlight the protective effect of D-JNKi in ischemia/reperfusion induced cell death of the retina.

Pankkien tuloksenjärjestelymahdollisuudet siirryttäessä IFRS-normistoon

Tutkielman ensisijaisena tavoitteena oli määrittää pankkien tuloksenjärjestelymahdollisuuksien muutoksia siirryttäessä kansallisesta FAS GAAP -tilinpäätösnormistosta IFRS-normistoon. Tuloksenjärjestelyn ilmiötä käsiteltiin esittelemällä useita koti- ja ulkomaisia tuloksenjärjestelytutkimuksia. Pankeilla on toimialalleen erityisiä kannustimia tuloksenjärjestelyyn. Pankkitoimialalla tuloksenjärjestely on perinteisesti esiintynyt tuloksentasaamisena tilikausien välillä. Tutkimuksessa saatiin selkeä kuva tuloksenjärjestelymahdollisuuksien muutoksesta. IFRS-normiston sisältämä johdon harkintavalta ilmenee tilinpäätöksen laadintaperiaatteiden valinnaisuutena ja standardien laskelmien sisältäminä johdon arvioina ja oletuksina. Harkintavaltaa käyttämällä johto voi vaikuttaa tilinpäätöksen taseeseen ja tulokseen. IFRS-normistossa tilinpäätöksen laadintaperiaatteisiin liittyvä johdon harkintavalta on vähentynyt verrattuna kansalliseen FAS GAAP-tilinpäätöskäytäntöön, mutta IFRS jättää johdolle edelleen harkintavaltaa muutamien kirjaus- ja raportointikäytäntöjen suhteen. Perinteisten tuloksenjärjestelykeinojen rinnalle IFRS:ssä nousevat useiden standardien laskelmien sisältämät johdon arviot ja oletukset tulevaisuuden muuttujista. Omaisuuserien käypään arvoon arvostamisen myötä tilikauden tuloksista saattaa tulla aikaisempaa volatiilimpia. Käypiin arvoihin arvostamisen seurauksena pankit saattavat siirtyä käyttämään sijoitustensa ostoja ja myyntejä tuloksentasauskeinona. FAS GAAP:ssa pankit ovat käyttäneet tuloksentasaamiseen tiettyjä suoriteperusteisia eriä, esimerkiksi luottotappiovarauksia. IFRS tiukentaa varausten tekemisen periaatteita, mutta tuo tilalle lainakannan arvonalentumistestaukseen sisältyvät johdon arviot.

Dual role of LldR in regulation of the lldPRD operon, involved in l-Lactate metabolism in Escherichia coli.

The lldPRD operon of Escherichia coli, involved in L-lactate metabolism, is induced by growth in this compound. We experimentally identified that this system is transcribed from a single promoter with an initiation site located 110 nucleotides upstream of the ATG start codon. On the basis of computational data, it had been proposed that LldR and its homologue PdhR act as regulators of the lldPRD operon. Nevertheless, no experimental data on the function of these regulators have been reported so far. Here we show that induction of an lldP-lacZ fusion by L-lactate is lost in an lldR mutant, indicating the role of LldR in this induction. Expression analysis of this construct in a pdhR mutant ruled out the participation of PdhR in the control of lldPRD. Gel shift experiments showed that LldR binds to two operator sites, O1 (positions 105 to 89) and O2 (positions 22 to 38), with O1 being filled at a lower concentration of LldR. L-Lactate induced a conformational change in LldR that did not modify its DNA binding activity. Mutations in O1 and O2 enhanced the basal transcriptional level. However, only mutations in O1 abolished induction by L-lactate. Mutants with a change in helical phasing between O1 and O2 behaved like O2 mutants. These results were consistent with the hypothesis that LldR has a dual role, acting as a repressor or an activator of lldPRD. We propose that in the absence of L-lactate, LldR binds to both O1 and O2, probably leading to DNA looping and the repression of transcription. Binding of L-lactate to LldR promotes a conformational change that may disrupt the DNA loop, allowing the formation of the transcription open complex.

Twist1 Is a TNF-Inducible Inhibitor of Clock Mediated Activation of Period Genes.

BACKGROUND: Activation of the immune system affects the circadian clock. Tumor necrosis factor (TNF) and Interleukin (IL)-1β inhibit the expression of clock genes including Period (Per) genes and the PAR-bZip clock-controlled gene D-site albumin promoter-binding protein (Dbp). These effects are due to cytokine-induced interference of E-box mediated transcription of clock genes. In the present study we have assessed the two E-box binding transcriptional regulators Twist1 and Twist2 for their role in cytokine induced inhibition of clock genes. METHODS: The expression of the clock genes Per1, Per2, Per3 and of Dbp was assessed in NIH-3T3 mouse fibroblasts and the mouse hippocampal neuronal cell line HT22. Cells were treated for 4h with TNF and IL-1β. The functional role of Twist1 and Twist2 was assessed by siRNAs against the Twist genes and by overexpression of TWIST proteins. In luciferase (luc) assays NIH-3T3 cells were transfected with reporter gene constructs, which contain a 3xPer1 E-box or a Dbp E-box. Quantitative chromatin immunoprecipitation (ChIP) was performed using antibodies to TWIST1 and CLOCK, and the E-box consensus sequences of Dbp (CATGTG) and Per1 E-box (CACGTG). RESULTS: We report here that siRNA against Twist1 protects NIH-3T3 cells and HT22 cells from down-regulation of Period and Dbp by TNF and IL-1β. Overexpression of Twist1, but not of Twist2, mimics the effect of the cytokines. TNF down-regulates the activation of Per1-3xE-box-luc, the effect being prevented by siRNA against Twist1. Overexpression of Twist1, but not of Twist2, inhibits Per1-3xE-box-luc or Dbp-E-Box-luc activity. ChIP experiments show TWIST1 induction by TNF to compete with CLOCK binding to the E-box of Period genes and Dbp. CONCLUSION: Twist1 plays a pivotal role in the TNF mediated suppression of E-box dependent transactivation of Period genes and Dbp. Thereby Twist1 may provide a link between the immune system and the circadian timing system.

Integrated regulation of the type III secretion system and other virulence determinants in Ralstonia solanacearum

In many plant and animal bacterial pathogens, the Type III secretion system (TTSS) that directly translocates effector proteins into the eukaryotic host cells is essential for the development of disease. In all species studied, the transcription of the TTSS and most of its effector substrates is tightly regulated by a succession of consecutively activated regulators. However, the whole genetic programme driven by these regulatory cascades is still unknown, especially in bacterial plant pathogens. Here, we have characterised the programme triggered by HrpG, a host-responsive regulator of the TTSS activation cascade in the plant pathogen Ralstonia solanacearum. We show through genome-wide expression analysis that, in addition to the TTSS, HrpG controls the expression of a previously undescribed TTSS-independent pathway that includes a number of other virulence determinants and genes likely involved in adaptation to life in the host. Functional studies revealed that this second pathway co-ordinates the bacterial production of plant cell wall-degrading enzymes, exopolysaccharide, and the phytohormones ethylene and auxin. We provide experimental evidence that these activities contribute to pathogenicity. We also show that the ethylene produced by R. solanacearum is able to modulate the expression of host genes and can therefore interfere with the signalling of plant defence responses. These results provide a new, integrated view of plant bacterial pathogenicity, where a common regulator activates synchronously upon infection the TTSS, other virulence determinants and a number of adaptive functions, which act co-operatively to cause disease.