Expression and characterization of two new alkane-inducible cytochrome P450s from Trichoderma harzianum

Abstract The inducibility CYPs by various carbon sources, including some n-alkanes and fatty acids, has been studied in Trichoderma harzianum. It was observed that n-dodecane and a mixture of fatty acids were good inducers of total CYP content and ω-hydroxylase of lauric acid, a marker for ω-hydroxylation of n-alkanes. By RACE it was isolated a cDNA containing an open reading frame of 1520 bp which encoded a CYP52 protein of 520 amino acids. Further, another n-alkane inducible CYP was identified in a library of T. harzianum by LC-MS/Ms analysis of a microsomal protein band induced by n-dodecane exposure. Thus, the filamentous fungus T. harzianum is expected to have a CYP dependent conversion of alkanes to fatty acids and their incorporation into cellular lipids

Effect of RNS60 in amyotrophic lateral sclerosis: a phase II multicentre, randomized, double-blind, placebo-controlled trial

Background and purpose Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease with limited treatment options. RNS60 is an immunomodulatory and neuroprotective investigational product that has shown efficacy in animal models of ALS and other neurodegenerative diseases. Its administration has been safe and well tolerated in ALS subjects in previous early phase trials. Methods This was a phase II, multicentre, randomized, double-blind, placebo-controlled, parallel-group trial. Participants diagnosed with definite, probable or probable laboratory-supported ALS were assigned to receive RNS60 or placebo administered for 24 weeks intravenously (375 ml) once a week and via nebulization (4 ml/day) on non-infusion days, followed by an additional 24 weeks off-treatment. The primary objective was to measure the effects of RNS60 treatment on selected biomarkers of inflammation and neurodegeneration in peripheral blood. Secondary objectives were to measure the effect of RNS60 on functional impairment (ALS Functional Rating Scale-Revised), a measure of self-sufficiency, respiratory function (forced vital capacity, FVC), quality of life (ALS Assessment Questionnaire-40, ALSAQ-40) and survival. Tolerability and safety were assessed. Results Seventy-four participants were assigned to RNS60 and 73 to placebo. Assessed biomarkers did not differ between arms. The mean rate of decline in FVC and the eating and drinking domain of ALSAQ-40 was slower in the RNS60 arm (FVC, difference 0.41 per week, standard error 0.16, p = 0.0101; ALSAQ-40, difference -0.19 per week, standard error 0.10, p = 0.0319). Adverse events were similar in the two arms. In a post hoc analysis, neurofilament light chain increased over time in bulbar onset placebo participants whilst remaining stable in those treated with RNS60. Conclusions The positive effects of RNS60 on selected measures of respiratory and bulbar function warrant further investigation

Effects of Global Warming on Patients with Dementia, Motor Neuron or Parkinson’s Diseases: A Comparison among Cortical and Subcortical Disorders

Exposure to global warming can be dangerous for health and can lead to an increase in the prevalence of neurological diseases worldwide. Such an effect is more evident in populations that are less prepared to cope with enhanced environmental temperatures. In this work, we extend our previous research on the link between climate change and Parkinson’s disease (PD) to also include Alzheimer’s Disease and other Dementias (AD/D) and Amyotrophic Lateral Sclerosis/Motor Neuron Diseases (ALS/MND). One hundred and eighty-four world countries were clustered into four groups according to their climate indices (warming and annual average temperature). Variations between 1990 and 2016 in the diseases’ indices (prevalence, deaths, and disability-adjusted life years) and climate indices for the four clusters were analyzed. Unlike our previous work on PD, we did not find any significant correlation between warming and epidemiological indices for AD/D and ALS/MND patients. A significantly lower increment in prevalence in countries with higher temperatures was found for ALS/MND patients. It can be argued that the discordant findings between AD/D or ALS/MND and PD might be related to the different features of the neuronal types involved and the pathophysiology of thermoregulation. The neurons of AD/D and ALS/MND patients are less vulnerable to heat-related degeneration effects than PD patients. PD patients’ substantia nigra pars compacta (SNpc), which are constitutively frailer due to their morphology and function, fall down under an overwhelming oxidative stress caused by climate warming

Proteome changes during yeast-like and pseudohyphal growth in the biofilm-forming yeast Pichia fermentans

The Pichia fermentans strain DISAABA 726 is a biofilm-forming yeast that has been proposed as biocontrol agent to control brown rot on apple. How ever, when inoculated on peach, strain 726 shows yeast-like to pseudohyphal transition coupled to a pathogenic behaviour. To identify the proteins potentially involved in such transition process, a comparative proteome analysis of P. fermentans 726 developed on peach (filamentous growth) vs apple (yeast-like growth) was carried out using two-dimensional gel electrophoresis coupled with mass spectrometry analysis. The proteome comparison was also performed between the two different cell morphologies induced in a liquid medium amended with urea (yeast-like cells) or methionine (filamentous cells) to exclude fruit tissue impact on the transition. Seventy-three protein spots showed significant variations in abundance (+/- twofold, p < 0.01, confidence intervals 99 %) between pseudohyphal vs yeast-like morphology produced on fruits. Among them, 30 proteins changed their levels when the two morphologies were developed in liquid medium. The identified proteins belong to several pathways and functions, such as glycolysis, amino acid synthesis, chaperones, and signalling transduction. The possible role of a group of proteins belonging to the carbohydrate pathway in the metabolic re-organisation during P. fermentans dimorphic transition is discussed

A Stress responsive alternative splicing mechanism in <i>Citrus clementina</i> leaves

Chitinases are often considered pathogenesis-related proteins since their activity can be induced by viral infections, fungal and bacterial cell wall components, and also by more general sources of stress such as wounding, salicylic acid, ethylene, auxins and cytokinins. In the present study, comparative proteomic analysis showed the defense-related acidic chitinase II to be specifically induced in Citrus clementina leaves infested by the two-spotted spider mite Tetranychus urticae or treated with MeJA. In parallel, changes in the mRNA profiles of two partially homologous chitinase forms were shown by RT-PCR. In particular, the appearance of an additional cDNA chitinase fragment in T. urticae-infested and MeJA-treated leaves was observed. This finding may indicate a specific regulatory mechanism of chitinase expression. We report evidence for alternative splicing in T. urticae-infested C. clementina, where a premature stop codon after the first 135 amino acids was introduced. We observed inducible chitinase activity after MeJA treatment, indicative of a rapid plant response to infestation. This work provides the first evidence of chitinase alternative splicing in C. clementina. In addition, the presence of the dual-band pattern for chitinase cDNA by RT-PCR may represent a suitable predictive marker for early diagnosis of plant biotic stress

Exosomal content in patients with ALS compared to those with selective involvement of lower motor neuron

The aim of our work was to study the exosomal specific content in ALS/MND patients, by comparing the exosomal-bound and free-circulating blood amounts of miR-155 and neuronal damage-asso- ciated proteins, and correlating them to blood cytokine level