Cultivable Bacteria Populations Associated with Leaves of Banana and Plantain Plants and Their Antagonistic Activity Against Mycosphaerella fijiensis

Mycosphaerella fijiensis is the etiological agent of Black Sigatoka, a fungal disease that affects production of banana and plantain crops in tropical regions. The sizes of cultivable epiphytic and endophytic bacterial populations, aerobic endospore forming bacteria (AEFB), and antagonist bacteria against M. fijiensis isolated from three Musa spp. cultivars from Urabá (Colombia) were studied, in order to find a suitable screening strategy to isolate antagonistic bacteria. Most of the variability found in the epiphytic and endophytic bacterial community sizes among fruit trees was explained by the cultivar differences. We found population sizes ranging from 1.25?×?10(3) to 9.64?×?10(5)?CFU/g of fresh leaf and found that 44?% of total cultivable bacteria belong to the AEFB group. We isolated 648 AEFB from three different cultivars and assessed their antagonistic activity against M. fijiensis using the cell-free supernatant obtained from bacterial liquid cultures in three different in vitro assays. Five percent of those bacteria showed higher percent inhibition than the positive control Bacillus subtilis UA321 has (percent inhibition?=?84?±?5) in the screening phase. Therefore, they were selected as antagonistic bacteria against the pathogen. The strains with the highest percentage of antagonism were found in older leaves for the three cultivars, given support to recommend this group of leaves for future samplings. Some of these isolated bacteria affected the mycelium and ascospores morphology of the fungus. They also presented in vitro characteristics related to a successful colonization of the phylloplane such as indolic compounds, surfactant production, and biofilm formation, which makes them possible, potential candidates as biological control agents.     

Analysis of natural variants of the hepatitis C virus internal ribosome entry site reveals that primary sequence plays a key role in cap-independent translation

The HCV internal ribosome entry site (IRES) spans a region of ~340 nt that encompasses most of the 5′ untranslated region (5′UTR) of the viral mRNA and the first 24–40 nt of the core-coding region. To investigate the implication of altering the primary sequence of the 5′UTR on IRES activity, naturally occurring variants of the 5′UTR were isolated from clinical samples and analyzed. The impact of the identified mutations on translation was evaluated in the context of RLuc/FLuc bicistronic RNAs. Results show that depending on their location within the RNA structure, these naturally occurring mutations cause a range of effects on IRES activity. However, mutations within subdomain IIId hinder HCV IRES-mediated translation. In an attempt to explain these data, the dynamic behavior of the subdomain IIId was analyzed by means of molecular dynamics (MD) simulations. Despite the loss of function, MD simulations predicted that mutant G266A/G268U possesses a structure similar to the wt-RNA. This prediction was validated by analyzing the secondary structure of the isolated IIId RNAs by circular dichroism spectroscopy in the presence or absence of Mg²⁺ ions. These data strongly suggest that the primary sequence of subdomain IIId plays a key role in HCV IRES-mediated translation.     

B subunit of Escherichia coli heat-labile enterotoxin as adjuvant of humoral immune response in recombinant BCG vaccination

The B subunit of Escherichia coli heat-labile enterotoxin (LTB), a nontoxic molecule with potent biological properties, is a powerful mucosal and parenteral adjuvant that induces a strong immune response against co-administered or coupled antigens. In this paper, the effect of LTB on the humoral immune response to recombinant BCG (rBCG) vaccination was evaluated. Isogenic mice were immunized with rBCG expressing the R1 repeat region of the P97 adhesin of Mycoplasma hyopneumoniae alone (rBCG/R1) or fused to LTB (rBCG/LTBR1). Anti-R1 systemic antibody levels (IgG1, IgG2a, IgG2b, IgG3, IgM, and IgA) were measured by ELISA using recombinant R1 as antigen. With the exception of IgM, LTB doubled the anti-R1 antibody levels in rBCG vaccination. The IgG1/IgG2a mean ratio showed that both rBCG/LTBR1 and rBCG/R1 induced a mixed Th1/Th2 immune response. Interestingly, anti-R1 serum IgA was induced only by rBCG/LTBR1. These results demonstrate that LTB has an adjuvant effect on the humoral immune response to recombinant antigens expressed in BCG.     

Multidrug resistance gene and P-glycoprotein expression in gastric adenocarcinoma and precursor lesions

Overexpression of the Multiple Drug Resistance gene (MDR1) has been proposed as a major mechanism related to both intrinsic and acquired resistance to chemotherapeutic agents. The gene product is a membrane protein (P-glycoprotein), that acts as an energydependent drug efflux pump decreasing drug accumulation in resistant tumor cells. We have characterized MDR1 and P-Glycoprotein expression in human gastric adenocarcinoma and in precursor lesions. MDR1 mRNAs, analyzed by dot-blot technique, were detected in 9 of 10 non-tumoral gastric mucosae and in 8 of 10 gastric adenocarcinomas. Immunohistochemical analysis, using the MRK16 monoclonal antibody, revealed heterogeneous expression of P-Glycoprotein in individual cells. The P-Glycoprotein was found on the surface of cells of gastric areas with intestinal metaplasia subtype III. This type of intestinal metaplasia, also called “colonic metaplasia”, has been strongly associated with a high risk for the development of gastric cancer. The fact that the P-Glycoprotein was detected in this precursor lesion is consistent with the intestinal metaplasia dysplasia and carcinoma sequence proposed in the histogenesis of this tumor. The finding that P-Glycoprotein was heterogeneously expressed in malignant cells of some gastric adenocarcinomas also suggests that this transporter system probably contributes to primary and secondary multidrug resistance in this neoplasm.     

Challenges for ecolabeling growth: lessons from the EU Ecolabel in Spain

The International Journal of Life Cycle Assessment - The European Ecolabel (EU Flower) has the mission to encourage cleaner production and influence consumers to promote Europe’s transition...     

Spider silks from plants – a challenge to create native-sized spidroins

Silk threads from spiders exhibit extraordinary mechanical properties, such as superior toughness and elasticity. Spider silks consist of several different large repetitive proteins that act as the basic materials responsible for these outstanding features. The production of spider silk protein variants in plants opens up new horizons in the production and functional investigation that enable the use of spider silks in innovative material development, nanotechnology and biomedicine in the future. This review summarizes and discusses production of spider silk protein variants in plants, especially with regards to plant expression systems, purification strategies, and characteristics of spider silk variants. Furthermore, the challenge of producing native-sized recombinant spidroins in planta is outlined, presenting three different strategies for achieving these high repetitive proteins with the help of non-repetitive C-terminal domains, crosslinking transglutaminase, and self-linking inteins. The potential of these fascinating proteins in medicine is also highlighted.