Current experimental disease-modifying therapeutics for multiple system atrophy.

Multiple system atrophy (MSA) is a challenging neurodegenerative disorder with a difficult and often inaccurate early diagnosis, still lacking effective treatment. It is characterized by a highly variable clinical presentation with parkinsonism, cerebellar ataxia, autonomic dysfunction, and pyramidal signs, with a rapid progression and an aggressive clinical course. The definite MSA diagnosis is only possible post-mortem, when the presence of distinctive oligodendroglial cytoplasmic inclusions (GCIs), mainly composed of misfolded and aggregated α-Synuclein (α-Syn) is demonstrated. The process of α-Syn accumulation and aggregation within oligodendrocytes is accepted one of the main pathological events underlying MSA. However, MSA is considered a multifactorial disorder with multiple pathogenic events acting together including neuroinflammation, oxidative stress, and disrupted neurotrophic support, among others. The discussed here treatment approaches are based on our current understanding of the pathogenesis of MSA and the results of preclinical and clinical therapeutic studies conducted over the last 2 decades. We summarize leading disease-modifying approaches for MSA including targeting α-Syn pathology, modulation of neuroinflammation, and enhancement of neuroprotection. In conclusion, we outline some challenges related to the need to overcome the gap in translation between preclinical and clinical studies towards a successful disease modification in MSA.

Histone Deacetylase 6 and the Disease Mechanisms of α-Synucleinopathies.

The abnormal accumulation of α-Synuclein (α-Syn) is a prominent pathological feature in a group of diseases called α-Synucleinopathies, such as Parkinson's disease, dementia with Lewy bodies (DLB), and multiple system atrophy (MSA). The formation of Lewy bodies (LBs) and glial cytoplasmic inclusions (GCIs) in neurons and oligodendrocytes, respectively, is highly investigated. However, the molecular mechanisms behind α-Syn improper folding and aggregation remain unclear. Histone deacetylase 6 (HDAC6) is a Class II deacetylase, containing two active catalytic domains and a ubiquitin-binding domain. The properties of HDAC6 and its exclusive cytoplasmic localization allow HDAC6 to modulate the microtubule dynamics, acting as a specific α-tubulin deacetylase. Also, HDAC6 can bind ubiquitinated proteins, facilitating the formation of the aggresome, a cellular defense mechanism to cope with higher levels of misfolded proteins. Several studies report that the aggresome shares similarities in size and composition with LBs and GCIs. HDAC6 is found to co-localize with α-Syn in neurons and in oligodendrocytes, together with other aggresome-related proteins. The involvement of HDAC6 in several neurodegenerative diseases is already under discussion, however, the results obtained by modulating HDAC6 activity are not entirely conclusive. The main goal of this review is to summarize and critically discuss previous in vitro and in vivo data regarding the specific role of HDAC6 in the context of α-Syn accumulation and protein aggregation in α-Synucleinopathies.

Targeting α-synuclein by PD03 AFFITOPE® and Anle138b rescues neurodegenerative pathology in a model of multiple system atrophy: clinical relevance.

BACKGROUND: Misfolded oligomeric α-synuclein plays a pivotal role in the pathogenesis of α-synucleinopathies including Parkinson's disease and multiple system atrophy, and its detection parallels activation of microglia and a loss of neurons in the substantia nigra pars compacta. Here we aimed to analyze the therapeutic efficacy of PD03, a new AFFITOPE® immunotherapy approach, either alone or in combination with Anle138b, in a PLP-α-syn mouse model. METHODS: The PLP-α-syn mice were treated with PD03 immunotherapy, Anle138b, or a combination of two. Five months after study initiation, the mice underwent behavioral testing and were sacrificed for neuropathological analysis. The treatment groups were compared to the vehicle group with regard to motor performance, nigral neuronal loss, microglial activation and α-synuclein pathology. RESULTS: The PLP-α-syn mice receiving the PD03 or Anle138b single therapy showed improvement of gait deficits and preservation of nigral dopaminergic neurons associated with the reduced α-synuclein oligomer levels and decreased microglial activation. The combined therapy with Anle138b and PD03 resulted in lower IgG binding in the brain as compared to the single immunotherapy with PD03. CONCLUSIONS: PD03 and Anle138b can selectively target oligomeric α-synuclein, resulting in attenuation of neurodegeneration in the PLP-α-syn mice. Both approaches are potential therapies that should be developed further for disease modification in α-synucleinopathies.

Effects of regular and decaffeinated roasted coffee (Coffea arabica and Coffea canephora) extracts and bioactive compounds on in vitro probiotic bacterial growth.

The aim of this study was to investigate the effects of coffee species, roast degree and decaffeination on in vitro probiotic bacterial growth, and to identify the major coffee compounds responsible for such effects. Six C. arabica and C. canephora extracts (regular medium and dark roasted and decaffeinated medium roasted), and five bioactive compounds (chlorogenic acid, galactomannan, type 2 arabinogalactan, caffeine and trigonelline) were individually incorporated into a modified low-carbon broth medium-(mMRS), at different concentrations (0.5 to 1.5% soluble coffee and 0.05 to 0.8 mg mL-1 standard solutions). Inulin and fructooligosaccharides (FOS) were used as prebiotic references. MRS and mMRS were used as rich and poor medium controls, respectively. The growth of Lactobacillus rhamnosus GG ATCC 53103-(GG), L. acidophilus LA-5-(LA), Bifidobacterium animalis DN-173010-(BA) and B. animalis subsp. lactis BB12-(BB12), as well as the growth inhibition of non-probiotic Escherichia coli ATCC 25922 were evaluated. Differences in growth between mMRS and treatments (Δlog CFU mL-1) were compared by ANOVA and Tukey's test, and considered when p ≤ 0.05. Overall, after 48 h incubation, the medium roasted arabica coffee extract increased the growth of GG, LA and BA (range: Δlog CFU mL-1 = 0.5 to 1.8), while the dark roasted arabica coffee extract increased BB12 growth (range: Δlog CFU mL-1 = 0.9 to 1.7), in a dose dependent manner. Improved performances of GG, LA and BA were promoted by higher polysaccharides and CGA concentrations, with better performance for Lactobacillus sp. The tested coffee bioactive compounds promoted the poor growth of BB12. Plain caffeine did not promote Bifidobacterium sp. growth and limited the growth of Lactobacillus sp. Regular C. arabica and C. canephora extracts inhibited the growth of E. coli, while the decaffeinated extracts promoted its growth. The present results show that coffee consumption can selectively improve the growth of probiotic strains, thus exerting a prebiotic effect, and show that coffee roasting and decaffeination affect this property and that different strains utilize different coffee components to grow.

Draft genome sequence of Enterococcus faecium E86, a strain producing broad-spectrum antimicrobial peptides: Description of a novel bacteriocin immunity protein and a novel sequence type.

OBJECTIVES: The aim of this study was to report the draft genome sequence of the bacteriocinogenic strain Enterococcus faecium E86. Bacteriocins are prokaryotic peptides or proteins with antimicrobial activity. The genome information may contribute to the identification of enterocins produced by this strain that exhibit inhibitory activity against the foodborne pathogen Listeria monocytogenes and vancomycin-resistant enterococci (VRE) involved in human infections, among other bacterial genera and species. METHODS: An Illumina MiSeq platform was used for genome sequencing. De novo assembly of 5 735 838 paired-end reads was done using the A5-miseq pipeline, yielding >300-fold average genome coverage. Genome annotation was performed by the RAST server, and mining of the bacteriocinogenic gene clusters was done using the BAGEL3 and antiSMASH v.4 platforms. RESULTS: The total scaffold size was determined to be 2 689 107 bp, approximately 2.7 Mbp, featuring a G + C content of 38.1%. The genome contains 2858 coding sequences and 74 RNA genes. Genome analyses revealed the presence of: 30 genes involved in drug resistance; 2 bacteriocinogenic gene clusters (for enterocin P and enterocin TW21); EntiTW21, a novel bacteriocin immunity protein and a novel multilocus sequence type (ST1500). CONCLUSION: This work highlights the potential biotechnological application of this strain for the production of enterocin P, a bacteriocin that can be employed in the food industry as a biopreservative against L. monocytogenes and as an alternative to classical antibiotics against VRE.

Automated NREM sleep staging using the electro-oculogram: a pilot study.

Automatic sleep staging from convenient and unobtrusive sensors has received considerable attention lately because this can enable a large range of potential applications in the clinical and consumer fields. In this paper the focus is on achieving non-REM (NREM) sleep staging from ocular electrodes. From these signals, specific patterns related to sleep such as slow eye movements, K-complexes, eye blinks, and spectral features are estimated. Although such patterns are characteristic of the Electroencephalogram, they can also be visible to a lesser extent on signals from ocular electrodes. Automatic sleep staging was implemented using two approaches: i) based on a state-machine and ii) using a neural network. The first one relied on the recommendations of the American Academy of Sleep Medicine, and the second one used a multilayer perceptron which was trained on manually sleep-staged data. Results were obtained on the data of five volunteers who participated in a nap experiment. Manual sleep staging of this data, performed by an expert, was used as reference. Five stages were considered, namely wake with eyes open, wake with eyes closed, and sleep stages N1, N2, and N3. The results were characterized in terms of confusion matrices from which the Cohen's κ coefficients were estimated. The values of κ for both the state-machine and neural-network based automatic sleep staging approaches were 0.79 and 0.59 respectively. Thus, the state-machine based approach shows a very good agreement with manual staging of sleep-data.

Inhibition of vancomycin and high-level aminoglycoside-resistant enterococci strains and Listeria monocytogenes by bacteriocin-like substance produced by Enterococcus faecium E86.

Three hundred and thirty nine lactic bacteria strains isolated from food samples were screened for antimicrobial activity. Only one strain isolated from meat pie and identified as Enterococcus faecium produced a bacteriocin-like inhibitory substance (BLIS) showing activity against Enterococcus, Leuconostoc, Lactobacillus, Listeria, Corynebacterium and Staphylococcus aureus. The BLIS produced was resistant to acid and alkali treatment and 121 masculineC for 15 min. The addition of BLIS in BHI contaminated with Listeria monocytogenes decreased the contamination in 4.8 log cycles in 24 h. The inhibition of listeria was also obtained in milk. Forty multiresistant enterococci strains were inhibited in the well-diffusion test. Two vancomycin resistant strains tested in liquid with BLIS were also inhibited. The BLIS producer showed no pathogenicity marker.