Pectinase production by <Emphasis Type="Italic">Aspergillus giganteus</Emphasis> in solid-state fermentation: optimization,scale-up,biochemical characterization and its application in olive-oil extraction

The application of pectinases in industrial olive-oil processes is restricted by its production cost. Consequently, new fungal strains able to produce higher pectinase titers are required. The aim of this work was to study the capability of Aspergillus giganteus NRRL10 to produce pectinolytic enzymes by SSF and evaluate the application of these in olive-oil extraction. A. giganteus was selected among 12 strains on the basis of high pectinolytic activity and stability. A mixture composed by wheat bran, orange, and lemon peels was selected as the best substrate for enzyme production. Statistical analyses of the experimental design indicated that pH, temperature, and CaCl₂ are the main factors that affect the production. Subsequently, different aeration flows were tested in a tray reactor; the highest activity was achieved at 20 L min^?1 per kilogram of dry substrate (kgds). Finally, the pectinolytic enzymes from A. giganteus improved the oil yield and rheological characteristics without affecting oil chemical properties.     

Strain engineering and process optimization for enhancing the production of a thermostable steryl glucosidase in <Emphasis Type="Italic">Escherichia coli</Emphasis>

Biodiesels produced from transesterification of vegetable oils have a major problem in quality due to the presence of precipitates, which are mostly composed of steryl glucosides (SGs). We have recently described an enzymatic method for the efficient removal of SGs from biodiesel, based on the activity of a thermostable β-glycosidase from Thermococcus litoralis. In the present work, we describe the development of an Escherichia coli-based expression system and a high cell density fermentation process. Strain and process engineering include the assessment of different promoters to drive the expression of a codon-optimized gene, the co-expression of molecular chaperones and the development of a high cell density fermentation process. A 200-fold increase in the production titers was achieved, which directly impacts on the costs of the industrial process for treating biodiesel.     

Frequency of a natural truncated allele of <Emphasis Type="Italic">MdMLO19</Emphasis> in the germplasm of <Emphasis Type="Italic">Malus domestica</Emphasis>

Podosphaera leucotricha is the causal agent of powdery mildew (PM) in apple. To reduce the amount of fungicides required to control this pathogen, the development of resistant apple cultivars should become a priority. Resistance to PM was achieved in various crops by knocking out specific members of the MLO gene family that are responsible for PM susceptibility (S-genes). In apple, the knockdown of MdMLO19 resulted in PM resistance. However, since gene silencing technologies such as RNAi are perceived unfavorably in Europe, a different approach that exploits this type of resistance is needed. This work evaluates the presence of non-functional naturally occurring alleles of MdMLO19 in apple germplasm. The screening of the re-sequencing data of 63 apple individuals led to the identification of 627 single nucleotide polymorphisms (SNPs) in five MLO genes (MdMLO5, MdMLO7, MdMLO11, MdMLO18, and MdMLO19), 127 of which were located in exons. The T-1201 insertion of a single nucleotide in MdMLO19 caused the formation of an early stop codon, resulting in a truncated protein lacking 185 amino acids, including the calmodulin-binding domain. The presence of the insertion was evaluated in 115 individuals. It was heterozygous in 64 and homozygous in 25. Twelve of the 25 individuals carrying the insertion in homozygosity were susceptible to PM. After barley, pea, cucumber, and tomato, apple would be the fifth species for which a natural non-functional mlo allele has been found.     

A lifelong competitive training practice attenuates age-related lipid peroxidation

The effect of exercise-induced oxidative stress on health and aging is not clearly explained. This study examined the effects of habitual sport practice, age, and submaximal exercise on the blood markers of oxidative stress, muscle damage, and antioxidant response. Seventy-two healthy men were grouped by their habitual sport practice: inactive (<1.5 h/week), recreational (3–8 h/week), and trained athletes (>8 h/week), and further divided by age: young (18–25 years), adult (40–55 years), and senior (>55 years). Blood samples were collected at rest and after submaximal effort. Hydroperoxides and superoxide dismutase, glutathione peroxidase, and catalase activities were measured by spectrophotometry. Nuclear DNA damage was analyzed by comet assay. The alpha-actin release was analyzed by Western blot. Alpha-tocopherol, retinol, and coenzyme-Q10 were quantified by high-performance liquid chromatography analysis. Data was analyzed through a factorial ANOVA and the Bonferroni post hoc test. Lipid peroxidation increased significantly with age and submaximal effort (p?<?0.05). However, the trained athlete group presented lower lipid peroxidation compared with the recreational group (MD?=?2.079, SED?=?0.58, p?=?0.002) and inactive group (MD?=?1.979, SED?=?0.61, p?=?0.005). Trained athletes showed significant higher alpha-actin levels (p?<?0.001) than the other groups. Recreational group showed lower nuclear DNA damage than trained athletes (MD?=?3.681, SED?=?1.28, p?=?0.015). Nevertheless, the inactive group presented significantly higher superoxide dismutase and catalase (p?<?0.05) than the other groups. Data suggested that habitual competitive training practice could prevent age-related increases of plasma lipid peroxidation, which, according with our results, cannot be entirely attributed to blood antioxidant defense systems.     

Functional proteomic insights in B-cell chronic lymphocytic leukemia

Introduction: B cell chronic lymphocytic leukemia (B-CLL) is a hematological malignancy considered as the most common leukemia in the Western world. The understanding of B cell differentiation is crucial for the diagnosis, prognosis, and treatment of the disease.

Areas covered: In this review, B-cell ontogeny and its relation with the CLL development, in combination with the proteomic approaches which could provide a deep characterization of the disease through the characterization of the cellular signaling pathways involved in the pathological cells is described.

Expert commentary: Although conventional strategies (genome sequencing, morphology assays, and immunophenotyping by flow cytometry and/or immunochemistry) have allowed the establishment of the disease stage based on different parameters, it is still necessary to utilize novel approaches (e.g., proteomics) that have the potential to simultaneously analyze thousands of molecules to improve understanding of CLL.     

Osmotic stress decreases complexity underlying the electrophysiological dynamic in soybean

• Studies on plant electrophysiology are mostly focused on specific traits of single cells. Inspired by the complexity of the signalling network in plants, and by analogy with neurons in human brains, we sought evidence of high complexity in the electrical dynamics of plant signalling and a likely relationship with environmental cues.
• An EEG‐like standard protocol was adopted for high‐resolution measurements of the electrical signal in Glycine max seedlings. The signals were continuously recorded in the same plants before and after osmotic stimuli with a ?2 MPa mannitol solution. Non‐linear time series analyses methods were used as follows: auto‐correlation and cross‐correlation function, power spectra density function, and complexity of the time series estimated as Approximate Entropy (ApEn).
• Using Approximate Entropy analysis we found that the level of temporal complexity of the electrical signals was affected by the environmental conditions, decreasing when the plant was subjected to a low osmotic potential. Electrical spikes observed only after stimuli followed a power law distribution, which is indicative of scale invariance.
• Our results suggest that changes in complexity of the electrical signals could be associated with water stress conditions in plants. We hypothesised that the power law distribution of the spikes could be explained by a self‐organised critical state (SOC) after osmotic stress.

     

Phylogenetic analysis of the genus <Emphasis Type="Italic">Argia</Emphasis> Rambur, 1842 (Odonata: Coenagrionidae), based on morphological characters of larvae and mitochondrial DNA sequences

The study of the evolutionary interrelationships among the species encompassed in the Neotropical genus Argia (Zygoptera: Coenagrionidae) has been neglected. The goal of this study is to infer the phylogenetic relationships among 36 species of Argia Rambur, 1842, using complementary data sets (i.e., larval morphology and mitochondrial DNA). The morphological data set comprises 76% of the larvae currently described for this genus and includes 97 morphological characters. From those, 47 characters have not been previously used in taxonomic studies involving dragonflies’ larvae. This is the first cladistic study based on larvae morphology for species within the suborder Zygoptera. Data partitions were analyzed individually, as well as total evidence, using parsimony and Bayesian inference as criteria for optimal-tree selection. The results support the monophyly of the North American species of Argia. This genus can be identified by the combination of eight synapomorphies, four of which are exclusively found in Argia. According to the optimal trees, the individual data sets (i.e., morphology and DNA sequences) have a high level of homoplasy, resulting in soft polytomies and low support for several nodes. The specific relationships of the terminal units differ between the phylogenies; nonetheless, there is historical congruence among them. Within Argia, five clades were consistently recovered. Most of those clades have been identified, at least in part, in previous phylogenetic and taxonomic studies. Indubitably, the morphological characters from larvae have historical signal useful for cladistic and taxonomic inference. Therefore, it should be a priority to pay more attention to this source of characters.     

Secondary biochemical and morphological consequences in lysosomal storage diseases

More than 50 hereditary lysosomal storage disorders (LSDs) are currently described. Most of these disorders are due to a deficiency of certain hydrolases/glycosidases and subsequent accumulation of nonhydrolyzable carbohydrate-containing compounds in lysosomes. Such accumulation causing hypertrophy of the lysosomal compartment is a characteristic feature of affected cells in LSDs. The investigation of biochemical and cellular parameters is of particular interest for understanding “life” of lysosomes in the normal state and in LSDs. This review highlights the wide spectrum of biochemical and morphological changes during developing LSDs that are extremely critical for many metabolic processes inside the various cells and tissues of affected persons. The data presented will help establish new complex strategies for metabolic correction of LSDs.     

VE和L-肌肽对大菱鲆幼鱼生长、抗氧化、非特异性免疫及血清生化指标的影响

实验以大菱鲆（Scophthalmus maximus）幼鱼[（14.00±0.02）g]为研究对象，采用2×4双因素设计，设2个VE水平（0和75 mg/kg）和4个L-肌肽水平（0、50、100和200 mg/kg），研究VE和L-肌肽对其生长、抗氧化、非特异性免疫及血清生化指标的影响。实验共分8组，每组3个重复，每个重复46尾鱼，实验周期为8周。结果显示:（1）在饲料中添加75 mg/kg VE显著提高了大菱鲆幼鱼增重率（WGR）和特定生长率（SGR）（P < 0.05），L-肌肽添加量≤100 mg/kg对实验鱼生长性能无显著影响（P>0.05），添加量为200 mg/kg时鱼体WGR、SGR和蛋白质效率（PER）显著降低，饲料系数（FCR）显著升高（P < 0.05）；（2）VE和L-肌肽对血清谷胱甘肽过氧化物酶（GSH-PX）、过氧化氢酶（CAT）活性和丙二醛（MDA）含量及肝脏总抗氧化能力（T-AOC）、超氧化物歧化酶（SOD）活性和MDA含量均具有显著的交互作用（P < 0.05），在VE 75 mg/kg水平下，L-肌肽添加量为50和100 mg/kg时血清GSH-PX活性最高，L-肌肽添加量为100和200 mg/kg时血清CAT活性最高且与添加量为50 mg/kg差异不显著（P>0.05），添加100 mg/kg肝脏T-AOC和SOD活性达到最高且50 mg/kg组的SOD与100 mg/kg组差异不显著（P>0.05），主效应结果显示，VE显著提高了血清T-AOC、SOD及肝脏CAT活性（P < 0.05），L-肌肽显著提高了血清T-AOC（P < 0.05）；（3）VE和L-肌肽对血清补体C3和LZM活性交互作用显著，在75 mg/kg VE水平下，L-肌肽添加量为50 mg/kg时，补体C3水平最高（P < 0.05），主效应显示，VE和L-肌肽对血清总蛋白（TP）影响均不显著（P>0.05）；（4）添加VE显著降低了血清总胆固醇（TCHO）和甘油三酯（TG）含量（P < 0.05），添加L-肌肽显著降低了血清TG含量，且在L-肌肽50 mg/kg时达到最低。综合考虑大菱鲆幼鱼[（14.00-39.43）g]的生长性能、抗氧化性能、非特异性免疫及血清生化指标得出，在实验配方条件下（鱼油70 g/kg，大豆卵磷脂10 g/kg），添加VE 75 mg/kg时，L-肌肽的适宜添加量为50 mg/kg。