Rationale and protocol of a double-blind, randomized, placebo-controlled trial to test the efficacy, safety, and tolerability of dimethyl fumarate in Friedreich Ataxia (DMF-FA-201)

IntroductionFriedreich Ataxia (FRDA) is an autosomal recessive neurodegenerative disorder that causes gait and limb ataxia, dysarthria, and impaired vibratory sense, with cardiomyopathy being the predominant cause of death. There is no approved therapy, which results in the use of symptomatic treatments and the chronic support of physiotherapy. Dimethyl fumarate (DMF) is a fumaric acid ester used for the treatment of psoriasis and Multiple Sclerosis (MS). It induces Nrf2 in vitro and in vivo, and it increases frataxin in FRDA patient lymphoblasts, in mouse models, and in MS treated patients.MethodsThe aim of our study is to investigate if DMF can increase the expression of the FXN gene and frataxin protein and ameliorate in-vivo detectable measures of mitochondrial dysfunction in FRDA. The study is composed of a screening visit and two sequential 12-week phases: a core phase and an extension phase. During the first phase (core), patients will be randomly assigned to either the DMF or a placebo group in a 1:1 ratio. During the first week, patients will receive a total daily dose of 240 mg of DMF or placebo; from the second week of treatment, the dose will be increased to two 120 mg tablets BID for a total daily dose of 480 mg. During the second phase (extension), all patients will be treated with DMF. EudraCT number 2021-006274-23.EndpointsThe primary endpoint will be a change in FXN gene expression level after 12 weeks of treatment. Secondary endpoints will be frataxin protein level, cardiopulmonary exercise test outputs, echocardiographic measures, Nrf2 pathway and mitochondrial biogenesis gene expression, safety, clinical scales, and quality of life scales.ConclusionsThis is the first study aimed at exploring the ability of DMF, an already available treatment for MS and psoriasis, to correct the biological deficits of FRDA and potentially improve mitochondrial respiration in-vivo

Biosynthesis of 2-Heptanone, a Volatile Organic Compound with a Protective Role against Honey Bee Pathogens, by Hive Associated Bacteria

Beehives are populated by bacterial species with a protective role against honey bee pathogens thanks to the production of bioactive metabolites. These compounds are largely unexploited despite their high potential interest for pest management. This study evaluated the capability of bacterial species associated with honey bees to produce 2-heptanone, a volatile organic compound with anesthetic properties of the parasitic mite Varroa destructor. The production of this compound was quantified by SPME-GC-MS in a culture filtrate of nine bacterial strains isolated from the surface of honey bees, and the biosynthetic potential was evaluated in bacterial species associated with apiaries by searching for protein homologs putatively involved in its biosynthesis by using biocomputational tools. The findings pointed out that 2-heptanone was produced by Acetobacteraceae bacterium, Bacillus thuringiensis and Apilactobacillus kunkeei isolates in concentrations between 1.5 and 2.6 ng/mL and that its production was strain-specific. Putative methylketone synthase homologs were found in Bacillus, Gilliamella, Acetobacteraceae, Bartonella and Lactobacillaceae, and the protein sequence results were distributed in nine Sequence Similarity Network (SSN) clusters. These preliminary results support the hypothesis that 2-heptanone may act as a mediator of microbial relationships in hives and provide contributions to assess the role and biosynthetic potential of 2-heptanone in apiaries

Emerging contaminants in agricultural ecosystems: impact of selected pharmaceutical on water and soil ecology and pratical implications

Pharmaceuticals are useful tools to prevent and treat human and animal diseases. Following administration, a significant fraction of pharmaceuticals is excreted unaltered into faeces and urine and may enter the aquatic ecosystem and agricultural soil through irrigation with recycled water, constituting a significant source of emerging contaminants into the environment. Understanding major factors influencing their environmental fate is consequently needed to value the risk, reduce contamination, and set up bioremediation technologies. The antiviral drug Tamiflu (oseltamivir carboxylate, OC) has received recent attention due to the potential use as a first line defence against H5N1 and H1N1 influenza viruses. Research has shown that OC is not removed during conventional wastewater treatments, thus having the potential to enter surface water bodies. A series of laboratory experiments investigated the fate and the removal of OC in surface water systems in Italy and Japan and in a municipal wastewater treatment plant. A preliminary laboratory study investigated the persistence of the active antiviral drug in water samples from an irrigation canal in northern Italy (Canale Emiliano Romagnolo). After an initial rapid decrease, OC concentration slowly decreased during the remaining incubation period. Approximately 65% of the initial OC amount remained in water at the end of the 36-day incubation period. A negligible amount of OC was lost both from sterilized water and from sterilized water/sediment samples, suggesting a significant role of microbial degradation. Stimulating microbial processes by the addition of sediments resulted in reduced OC persistence. Presence of OC (1.5 μg mL-1) did not significantly affect the metabolic potential of the water microbial population, that was estimated by glyphosate and metolachlor mineralization. In contrast, OC caused an initial transient decrease in the size of the indigenous microbial population of water samples. A second laboratory study focused on basic processes governing the environmental fate of OC in surface water from two contrasting aquatic ecosystems of northern Italy, the River Po and the Venice Lagoon. Results of this study confirmed the potential of OC to persist in surface water. However, the addition of 5% of sediments resulted in rapid OC degradation. The estimated half-life of OC in water/sediment of the River Po was 15 days. After three weeks of incubation at 20 °C, more than 8% of 14C-OC evolved as 14CO2 from water/sediment samples of the River Po and Venice Lagoon. OC was moderately retained onto coarse sediments from the two sites. In water/sediment samples of the River Po and Venice Lagoon treated with 14C-OC, more than 30% of the 14C-residues remained water-extractable after three weeks of incubation. The low affinity of OC to sediments suggests that the presence of sediments would not reduce its bioavailability to microbial degradation. Another series of laboratory experiments investigated the fate and the removal of OC in two surface water ecosystems of Japan and in the municipal wastewater treatment plant of the city of Bologna, in Northern Italy. The persistence of OC in surface water ranged from non-detectable degradation to a half-life of 53 days. After 40 days, less than 3% of radiolabeled OC evolved as 14CO2. The presence of sediments (5%) led to a significant increase of OC degradation and of mineralization rates. A more intense mineralization was observed in samples of the wastewater treatment plant when applying a long incubation period (40 days). More precisely, 76% and 37% of the initial radioactivity applied as 14C-OC was recovered as 14CO2 from samples of the biological tank and effluent water, respectively. Two bacterial strains growing on OC as sole carbon source were isolated and used for its removal from synthetic medium and environmental samples, including surface water and wastewater. Inoculation of water and wastewater samples with the two OC-degrading strains showed that mineralization of OC was significantly higher in both inoculated water and wastewater, than in uninoculated controls. Denaturing gradient gel electrophoresis and quantitative PCR analysis showed that OC would not affect the microbial population of surface water and wastewater. The capacity of the ligninolytic fungus Phanerochaete chrysosporium to degrade a wide variety of environmentally persistent xenobiotics has been largely reported in literature. In a series of laboratory experiments, the efficiency of a formulation using P. chrysosporium was evaluated for the removal of selected pharmaceuticals from wastewater samples. Addition of the fungus to samples of the wastewater treatment plant of Bologna significantly increased (P < 0.05) the removal of OC and three antibiotics, erythromycin, sulfamethoxazole, and ciprofloxacin. Similar effects were also observed in effluent water. OC was the most persistent of the four pharmaceuticals. After 30 days of incubation, approximately two times more OC was removed in bioremediated samples than in controls. The highest removal efficiency of the formulation was observed with the antibiotic ciprofloxacin. The studies included environmental aspects of soil contamination with two emerging veterinary contaminants, such as doramectin and oxibendazole, wich are common parasitic treatments in cattle farms

Biosynthesis of 2-Heptanone, a Volatile Organic Compound with a Protective Role against Honey Bee Pathogens, by Hive Associated Bacteria

Beehives are populated by bacterial species with a protective role against honey bee pathogens thanks to the production of bioactive metabolites. These compounds are largely unexploited despite their high potential interest for pest management. This study evaluated the capability of bacterial species associated with honey bees to produce 2-heptanone, a volatile organic compound with anesthetic properties of the parasitic mite Varroa destructor. The production of this compound was quantified by SPME-GC-MS in a culture filtrate of nine bacterial strains isolated from the surface of honey bees, and the biosynthetic potential was evaluated in bacterial species associated with apiaries by searching for protein homologs putatively involved in its biosynthesis by using biocomputational tools. The findings pointed out that 2-heptanone was produced by Acetobacteraceae bacterium, Bacillus thuringiensis and Apilactobacillus kunkeei isolates in concentrations between 1.5 and 2.6 ng/mL and that its production was strain-specific. Putative methylketone synthase homologs were found in Bacillus, Gilliamella, Acetobacteraceae, Bartonella and Lactobacillaceae, and the protein sequence results were distributed in nine Sequence Similarity Network (SSN) clusters. These preliminary results support the hypothesis that 2-heptanone may act as a mediator of microbial relationships in hives and provide contributions to assess the role and biosynthetic potential of 2-heptanone in apiaries

Changes in rhizosphere bacterial communities associated to tree decline: grapevine esca syndrome case study

An investigation was carried out on rhizosphere bacteria to determine if they may be associated with perennial crops affected by nonspecific decline, a phenomenon that is difficult to diagnose and prevent. Esca disease of grapevine was chosen for this case study because of its easy foliar symptom identification. Ribosomal DNA fingerprint analysis by polymerase chain reaction – denaturing gradient gel electrophoresis (PCR–DGGE), quantitative PCR (qPCR), and rDNA amplicon sequencing by next-generation sequencing (NGS) were adopted to investigate the bacterial communities associated with grapevines, which were selected for the presence and absence of external foliar symptoms in 11 vineyards. According to PCR–DGGE and qPCR, bacterial communities differed in site of origin (vineyards), but not between symptomatic and asymptomatic plants, whereas qPCR gave a significantly higher presence of total bacteria and Pseudomonas spp. in asymptomatic plants. NGS confirmed no difference between symptomatic and asymptomatic plants, apart from a few minor genera (<0.5%) such as Salinibacterium, Flavobacterium, Nocardia, and Janthinobacterium, which were, in all cases, higher in asymptomatic plants and whose functional role should be the object of further investigation. The fact that total bacteria and Pseudomonas were more abundant in the rhizosphere of asymptomatic grapevines and that some bacterial genera were associated with the latter, represents a new element when investigating the multiple-origin phenomenon such as esca disease of grapevine.The accepted manuscript in pdf format is listed with the files at the bottom of this page. The presentation of the authors' names and (or) special characters in the title of the manuscript may differ slightly between what is listed on this page and what is listed in the pdf file of the accepted manuscript; that in the pdf file of the accepted manuscript is what was submitted by the author

Removal of oseltamivir (Tamiflu) and other selected pharmaceuticals from wastewater using a granular bioplastic formulation entrapping propagules of Phanerochaete chrysosporium.

International audienceThe capacity of the ligninolytic fungus Phanerochaete chrysosporium to degrade a wide variety of environmentally persistent xenobiotics has been largely reported in the literature. Beside other factors, one barrier to a wider use of this bioremediation fungus is the availability of effective formulations that ensure easy preparation, handling and application. In this series of laboratory experiments, we evaluated the efficiency of a granular bioplastic formulation entrapping propagules of P. chrysosporium for removal of four selected pharmaceuticals from wastewater samples. Addition of inoculated granules to samples of the wastewater treatment plant of Bologna significantly increased the removal of the antiviral drug oseltamivir (Tamiflu), and the antibiotics, erythromycin, sulfamethoxazol, and ciprofloxacin. Similar effects were also observed in effluent water. Oseltamivir was the most persistent of the four active substances. After 30d of incubation, approximately two times more oseltamivir was removed in bioremediated wastewater than controls. The highest removal efficiency of the bioplastic formulation was observed with the antibiotic ciprofloxacin. Microbiological DNA-based analysis showed that the bioplastic matrix supported the growth of P. chrysosporium, thus facilitating its adaptation to unusual environment such as wastewater

Molecular stress responses to nano-sized zero-valent iron (nZVI) particles in the soil bacterium Pseudomonas stutzeri.

Nanotoxicological studies were performed in vitro using the common soil bacterium Pseudomonas stutzeri to assess the potentially toxic impact of commercial nano-sized zero-valent iron (nZVI) particles, which are currently used for environmental remediation projects. The phenotypic response of P. stutzeri to nZVI toxicity includes an initial insult to the cell wall, as evidenced by TEM micrographs. Transcriptional analyses using genes of particular relevance in cellular activity revealed that no significant changes occurred among the relative expression ratios of narG, nirS, pykA or gyrA following nZVI exposure; however, a significant increase in katB expression was indicative of nZVI-induced oxidative stress in P. stutzeri. A proteomic approach identified two major defence mechanisms that occurred in response to nZVI exposure: a downregulation of membrane proteins and an upregulation of proteins involved in reducing intracellular oxidative stress. These biomarkers served as early indicators of nZVI response in this soil bacterium, and may provide relevant information for environmental hazard assessment

The Role of the Bacterial Community of an Agroecosystem in Simazine Degradation

The use of pesticides and fertilizers in agricultural practice is the main source of soil and groundwater contamination. S-Triazines are among the most used herbicides in the world for selective weed control in several types of crops. The homeostatic capability of an agroecosystem to remove a triazinic herbicide, simazine, was assessed in microcosms treated with the herbicide in presence/absence of urea fertilizer. The latter, as well as a fertilizer, is also one of the last by-products before simazine mineralization. The biodegradation, in terms of disappearance of 50% of the initial concentration (DT50), was compared to the degradation and metabolite formation occurring in sterilized soil. Moreover, the bacterial community response was assessed in terms of abundance and community structure by the epifluorescence direct count method and fluorescence in situ hybridization. The results show that the microbial community has a primary role in simazine degradation and that this process is due to the presence of a microbial pool working in succession and of which the metabolism may be modulated by exogenous sources of nitrogen, like urea. The latter influences the degradative pathway with a greater formation and accumulation of the desethyl-simazine metabolite, which is a hazardous contaminant of soil and groundwater ecosystems, as well as its parent compound

The relationship between tree species and wood colonising fungi and fungal interactions influences wood degradation

Deadwood is one of the main terrestrial carbon (C) pools and its decomposition is fundamental in biogeochemical cycles. As fungi are the main wood degraders, a study was carried out on deadwood-colonising fungal community changes as a function of host plants and wood degradation. Deadwood fungal communities were investigated in adjacent woody plantations of hybrid poplar, willow and black locust established in the early 2000 s and abandoned for fifteen years. Lying deadwood was estimated in the three plantations and wood decay (WD) visually assessed using a 3-class scale. Fungal colonization of wood samples was assessed using culture-based methods and identified with morphological and molecular features. Wood colonising fungal communities differed in composition both between host plants and WD classes (P < 0.001). Ligninolytic white rot fungal species (Basidiomycota) prevailed in black locust and poplar, where they co-colonised all three wood decay classes in association with soft rot fungal agents. In willow, Daldinia chilidiae, a ligninolytic ascomycete, co-colonised deadwood with fungal species associated with tree dieback (Botryosphaeriaceae); whilst soft rot fungi principally colonised the highest wood degradation class. Results showed high fungal preference towards tree species that was most evident in ligninolytic fungi. Moreover, different wood decay paths were deduced from fungal changes across the three wood degrading classes and the necromass distribution in those classes. The co-habitation of white rot and soft rot fungi in all the wood decay classes of poplar and black locust indicated a continuum decay process; whilst the prevalence of soft rot fungi in the almost decomposed class in willow, suggested sequential succession of primary and secondary phases in deadwood degradation. Wood degradation is influenced by plant species and the native fungal microbiome and is mediated by multiple biotic interactions and environmental factors, which supports the use of wood colonising fungi as indicators of forest ecosystem health

TEM micrographs showing <i>P. stutzeri</i> control cells (A) and cells exposed to 5 g L⁻¹ nZVI (B).

<p>TEM micrographs showing <i>P. stutzeri</i> control cells (A) and cells exposed to 5 g L⁻¹ nZVI (B).</p