Plant-microbe interactions and the new biotechnological methods of plant disease control

Plants constitute an excellent ecosystem for microorganisms. The environmental conditions offered differ considerably between the highly variable aerial plant part and the more stable root system. Microbes interact with plant tissues and cells with different degrees of dependence. The most interesting from the microbial ecology point of view, however, are specific interactions developed by plant-beneficial (either non-symbiotic or symbiotic) and pathogenic microorganisms. Plants, like humans and other animals, also become sick, but they have evolved a sophisticated defense response against microbes, based on a combination of constitutive and inducible responses which can be localized or spread throughout plant organs and tissues. The response is mediated by several messenger molecules that activate pathogen-responsive genes coding for enzymes or antimicrobial compounds, and produces less sophisticated and specific compounds than immunoglobulins in animals. However, the response specifically detects intracellularly a type of protein of the pathogen based on a gene-for-gene interaction recognition system, triggering a biochemical attack and programmed cell death. Several implications for the management of plant diseases are derived from knowledge of the basis of the specificity of plant-bacteria interactions. New biotechnological products are currently being developed based on stimulation of the plant defense response, and on the use of plant-beneficial bacteria for biological control of plant diseases (biopesticides) and for plant growth promotion (biofertilizers)

Identification of valid reference genes during the differentiation of human myoblasts

<p>Abstract</p> <p>Background</p> <p>Analysis of RNA expression using real-time PCR (qRT-PCR) traditionally includes reference genes (RG) as an internal control. This practice is being questioned as it becomes increasingly clear that RG may vary considerably under certain experimental conditions. Thus, the validity of a particular RG must be determined for each experimental setting. We used qRT-PCR to measure the levels of six RG, which have been reported in the literature to be invariant. The RG were analyzed in human myoblast cultures under differentiation conditions. We examined the expression by qRT-PCR of mRNA encoding Beta-actin (ACTB), Beta-2-microglobulin (B2M), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), peptidylprolyl isomerase A (PPIA), TATA box binding protein (TBP) and ribosomal protein, large, P0 (RPLPO). The mRNA expression of the following genes of interest (GOI) were analyzed: skeletal muscle alpha 1 actin (ACTA1), myogenin/myogenic factor 4 (MYOG), embryonic skeletal muscle myosin heavy chain 3 (MYH3) and the activity of creatine phosphokinase (CK). The geNorm, NormFinder and BestKeeper software programs were used to ascertain the most suitable RG to normalize the RNA input.</p> <p>Results</p> <p>Using the geNorm program, RPLPO and TBP were found to be the most stable genes, additionally a suitable normalization factor (NF) was calculated. The NormFinder software showed that RPLPO was the most stable, whereas TBP ranked second. BestKeeper program also revealed that RPLPO and TBP as stable genes, but PPIA was the most stable reference gene, whereas GAPDH and ACTB were the worst ranked.</p> <p>Conclusion</p> <p>RNA expression analyses including three independent softwares revealed that RPLPO, TBP as reference genes or NF calculated by geNorm software, are suitable to normalize the mRNA expression in myoblast after culture under differentiation conditions. Significant correlations can be identified between the differentiations markers ACTA1, MYOG, MYH3 and creatine phosphokinase (CK) activity, when the expression is normalized with the NF calculated with RPLPO and TBP.</p

Phytohustil and root extract of Althaea officinalis L. exert anti-inflammatory and anti-oxidative properties and improve the migratory capacity of endothelial cells in vitro

Introduction: Althaea officinalis L.'s root extract (REA) has been used as a medicinal plant since ancient times to treat a cough. Applying REA leads to a protective film that induces a faster regeneration of the lesioned laryngopharyngeal mucosa caused by dry coughs. The buccopharyngeal mucosa is a highly vascularized tissue. In this regard, anti-inflammatory/-oxidant phytochemicals that improve the repair of the lesion site, e.g., neovascularization in the wound, are critical for promoting healing. For this reason, it is essential to investigate the effects of Phytohustil® and REA on different cellular components of the mucosa under conditions similar to those found in the injured mucosa. Thus, this in vitro study investigated the anti-inflammatory/oxidative and pro-migratory properties of Phytohustil® cough syrup on vascular endothelial cells. Methods: Human umbilical vein endothelial cells (HUVEC) were pretreated (24 h) with Phytohustil®, its excipients, or REA, followed by incubation with hydrogen peroxide (H2O2; 1 h; pro-oxidative) or with lipopolysaccharides (LPS; 3 h; pro-inflammatory). Viability and cytotoxicity were measured by PrestoBlue® assay. Intracellular reactive oxygen species (ROS) were quantified with 20-70-dichlorofluorescein diacetate (DCFDA). The release of interleukin 6 (IL6) was determined by enzyme-linked immunosorbent assay (ELISA). The migratory capacity of HUVEC was measured using a scratch assay. Results: Our results show that Phytohustil®, its excipients and REA were not cytotoxic. Pretreatment of HUVEC (24 h) with Phytohustil® or REA inhibited the LPS-activated IL6 release. Phytohustil® or REA inhibited the H2O2-induced cytotoxicity and intracellular ROS production. Phytohustil® and REA significantly stimulated wound closure compared to the control. Conclusion: Our data show that Phytohustil® and REA have anti-inflammatory/-oxidant properties and improve the migratory capacity of vascular endothelial cells. These properties may contribute to the healing characteristics of Phytohustil® and support the benefit of Phytohustil® in patient’s treatment of irritated oral mucosa

Increased Density of Growth Differentiation Factor-15+ Immunoreactive M1/M2 Macrophages in Prostate Cancer of Different Gleason Scores Compared with Benign Prostate Hyperplasia

Prostate cancer (PCa) is the second most diagnosed cancer and cause of death in men worldwide. The main challenge is to discover biomarkers for malignancy to guide the physician towards optimized diagnosis and therapy. There is recent evidence that growth differentiation factor-15 (GDF-15) is elevated in cancer patients. Therefore, we aimed to decipher GDF-15+ cell types and their density in biopsies of human PCa patients with Gleason score (GS)6–9 and benign prostate hyperplasia (BPH). Here we show that the density of GDF-15+ cells, mainly identified as interstitial macrophages (MΦ), was higher in GS6–9 than in BPH, and, thus, GDF-15 is intended to differentiate patients with high GS vs. BPH, as well as GS6 vs. GS7 (or even with higher malignancy). Some GDF-15+ MΦ showed a transepithelial migration into the glandular lumen and, thus, might be used for measurement in urine/semen. Taken together, GDF-15 is proposed as a novel tool to diagnose PCa vs. BPH or malignancy (GS6 vs. higher GS) and as a potential target for anti-tumor therapy. GDF-15 in seminal plasma and/or urine could be utilized as a non-invasive biomarker of PCa as compared to BPH

Aplicación de Aproximantes de Padé en el Modelado de la Viscosidad de Jarabe de Maíz

The industrialization of corn is one of the Argentina’s agro-industrial activities that generate higher value added, allowing to obtain a large number of products intended for direct consumption or are used as inputs in other industries. This paper proposes the application of Padé approximants as an alternative method&nbsp;in&nbsp;the search for new&nbsp;nonlinear models that&nbsp;successfully&nbsp;describe&nbsp;the relationships between&nbsp;the different parameters involved in an algebraic expression to estimate food properties. Is considered for study the viscosity of corn syrup. Objectives are established to correlate experimental information available about properties of corn syrup, Padé approximants applied in modeling a general expression and to verify and validate the model against experimental data. As a result is observed an adequate fit to the experimental data withaverage absolute deviations less than 6% and high values of R2La industrialización del maíz es una de las actividades agroindustriales de Argentina que genera mayor valor agregado, permitiendo obtener un gran número de productos que se destinan al consumo directo o bien son empleados como insumos en otras industrias. En este trabajo se propone la aplicación de aproximantes de Padé como un método alternativo en la búsqueda de nuevos modelos no lineales que describan satisfactoriamente las relaciones existentes entre los distintos parámetros involucrados en una expresión algebraica, para estimar propiedades de alimentos. Se considera para su estudio la viscosidad de jarabe de maíz. Se establecen como objetivos correlacionar información experimental disponible sobre propiedades de jarabe de maíz, aplicar aproximantes de Padé en la modelación de una expresión general y comprobar y validar el modelo frente a datos experimentales. Como resultado se observa un ajuste adecuado a los datos experimentales, con desviaciones absolutas promedio menores a 6% y valores elevados de R

Data on autophagy markers in human macrophages exposed to oxLDL and growth differentiation factor-15

Growth differentiation factor-15 (GDF-15) is a member of the TGF-β superfamily, identical to MΦ-inhibitory cytokine-1 (MIC-1). GDF-15 is associated with e.g. cardiovascular disease, inflammation and development of atherosclerosis and is highly expressed in macrophages (MΦ) of atherosclerotic lesions. Moreover, there exists an indication for the involvement of oxidized-low density lipoprotein (oxLDL) uptake and autophagic processes by MΦ regarding arteriosclerotic progression. Thus, we were interested to investigate a potential regulatory effect of GDF-15 on autophagy signaling pathway in human MΦ during foam cell formation. Here, we present western blot data of ATG5, ATG12/ATG5-complex and p62 regarding the GDF-15 concentration. For further interpretation of the data presented in this article, please see the research article “Growth differentiation factor-15 regulates oxLDL-induced lipid homeostasis and autophagy in human macrophages” [1]