Neuronal control of microglia through the mitochondria.

The microbial toxin ß-N-methylamino-L-alanine (BMAA), which is derived from cyanobacteria, targets neuronal mitochondria, leading to the activation of neuronal innate immunity and, consequently, neurodegeneration. Although known to modulate brain inflammation, the precise role of aberrant microglial function in the neurodegenerative process remains elusive. To determine if neurons signal microglial cells, we treated primary cortical neurons with BMAA and then co-cultured them with the N9 microglial cell line. Our observations indicate that microglial cell activation requires initial neuronal priming. Contrary to what was observed in cortical neurons, BMAA was not able to activate inflammatory pathways in N9 cells. We observed that microglial activation is dependent on mitochondrial dysfunction signaled by BMAA-treated neurons. In this scenario, the NLRP3 pro-inflammatory pathway is activated due to mitochondrial impairment in N9 cells. These results demonstrate that microglia activation in the presence of BMAA is dependent on neuronal signaling. This study provides evidence that neurons may trigger microglia activation and subsequent neuroinflammation. In addition, we demonstrate that microglial activation may have a protective role in ameliorating neuronal innate immune activation, at least in the initial phase. This work challenges the current understanding of neuroinflammation by assigning the primary role to neurons.

What is the role of PEO chains in the assembly of core-corona supraparticles in aqueous dispersions?

Hypothesis The assembly of core-corona supraparticles in aqueous dispersions has been regularly assisted by auxiliary monomers/oligomers which modify the individual particles with, e.g., surface grafting of polyethylene oxide (PEO) chains or other hydrophilic monomers. However, this modification complicates the preparation and purification procedures and increases potential upscaling efforts. Hybrid polymer-silica core-corona supracolloids could be more simply assembled if the PEO chains from surfactants, typically used by default as polymer stabilizers, concomitantly act as assembly promotors. The supracolloids assembly could therefore be more easily achieved without requiring particles functionalization or post-purification steps. Methods The self-assembly of supracolloidal particles prepared with PEO-surfactant stabilized (Triton X-405) and/or PEO-grafted polymer particles is compared to differentiate the roles of the PEO chains in the assembly of core-corona supraparticles. Using time-resolved dynamic light scattering (DLS) and cryogenic transmission electron microscopy(cryo-TEM), the effect of concentration of PEO chains (from surfactant) on the kinetics and dynamics of supracolloids assembly is investigated. Self-consistent field (SCF) lattice theory was used to numerically study the distribution of PEO chains at the interfaces present in the supracolloidal dispersions. Findings The PEO based surfactant can be used as assembly promoter of core-corona hybrid supracolloids due to its amphiphilic nature and via establishing hydrophobic interactions. The concentration of the PEO surfactant, and especially the PEO chains distribution over the different interfaces, crucially affect the supracolloids assembly. A simplified pathway for preparing hybrid supracolloidal particles with a well-controlled corona coverage over polymer cores is presented.

Tunable distribution of silica nanoparticles in water-borne coatings via strawberry supracolloidal dispersions.

HYPOTHESIS: Water-borne coatings are rapidly expanding as sustainable alternatives to organic solvent-borne systems. Inorganic colloids are often added to aqueous polymer dispersions to enhance the performance of water-borne coatings. However, these bimodal dispersions have many interfaces which can result in unstable colloids and undesirable phase separation. The covalent bonding between individual colloids, on a polymer-inorganic core-corona supracolloidal assembly, could reduce or suppress instability and phase separation during drying of coatings, advancing its mechanical and optical properties. METHODS: Aqueous polymer-silica supracolloids with a core-corona strawberry configuration were used to precisely control the silica nanoparticles distribution within the coating. The interaction between polymer and silica particles was fine-tuned to obtain covalently bound or physically adsorbed supracolloids. Coatings were prepared by drying the supracolloidal dispersions at room temperature, and their morphology and mechanical properties were interconnected. FINDINGS: Covalently bound supracolloids provided transparent coatings with a homogeneous 3D percolating silica nanonetwork. Supracolloids having physical adsorption only, resulted in coatings with a stratified silica layer at interfaces. The well-arranged silica nanonetworks strongly improve the storage moduli and water resistance of the coatings. These supracolloidal dispersions offer a new paradigm for preparing water-borne coatings with enhanced mechanical properties and other functionalities, like structural color.

Intestinal infection triggers mitochondria-mediated α-synuclein pathology: relevance to Parkinson's disease.

Parkinson's disease (PD) is a multifactorial neurodegenerative disease characterized by the loss of dopaminergic neurons in the midbrain. In the prodromal phase several autonomic symptoms including orthostatic hypotension and constipation are correlated with increased α-synuclein pathology in peripheral tissues. It is currently accepted that some idiopathic PD cases may start in the gut (body-first PD) with accumulation of pathological α-synuclein in enteric neurons that may subsequently propagate caudo-rostrally to the central nervous system. In addition to the already-established regulation of synaptic vesicle trafficking, α-synuclein also seems to play a role in neuronal innate immunity after infection. Our goal was to understand if seeding the gut with the foodborne pathogen Listeria monocytogenes by oral gavage would impact gut immunity and eventually the central nervous system. Our results demonstrate that L. monocytogenes infection induced oligomerization of α-synuclein in the ileum, along with a pronounced pro-inflammatory local and systemic response that ultimately culminated in neuronal mitochondria dysfunction. We propose that, having evolved from ancestral endosymbiotic bacteria, mitochondria may be directly targeted by virulence factors of intracellular pathogens, and that mitochondrial dysfunction and fragmentation resulting also from the activation of the innate immune system at the gut level, trigger innate immune responses in midbrain neurons, which include α-synuclein oligomerization and neuroinflammation, all of which hallmarks of PD.

LPS-induced mitochondrial dysfunction regulates innate immunity activation and α-synuclein oligomerization in Parkinson's disease.

Sporadic Parkinson's disease (sPD) is a complex multifactorial disorder which etiology remains elusive. Several mechanisms have been described to contribute to PD development namely mitochondrial dysfunction, activation of inflammatory pathways and the deposition of unfolded proteins such as α-synuclein. Our work shows for the first time that lipopolysaccharide (LPS)-induced activation of innate immunity requires a functional mitochondria and mimics PD pathology in cells. We found in primary mesencephalic neurons that LPS targeted the mitochondria and activated neuronal innate immune responses, which culminated with α-synuclein oligomerization. Moreover, in cybrid cell lines repopulated with mtDNA from sPD subjects with inherent mitochondrial dysfunction and NT2-Rho0 obtained by long-term ethidium bromide exposure, and so without a functional mitochondrial, LPS was not able to further activate innate immunity or increase α-synuclein aggregation. Herein, we showed that mesencephalic neurons are able to activate innate immunity after LPS exposure and this pathway is dependent on mitochondria. Moreover, we disclose that α-synuclein over production is an innate immune response. Our data indicate that mitochondria provide the base for innate immunity activation in idiopathic PD.

Footprints of a microbial toxin from the gut microbiome to mesencephalic mitochondria.

OBJECTIVE: Idiopathic Parkinson's disease (PD) is characterised by alpha-synuclein (aSyn) aggregation and death of dopaminergic neurons in the midbrain. Recent evidence posits that PD may initiate in the gut by microbes or their toxins that promote chronic gut inflammation that will ultimately impact the brain. In this work, we sought to demonstrate that the effects of the microbial toxin ß-N-methylamino-L-alanine (BMAA) in the gut may trigger some PD cases, which is especially worrying as this toxin is present in certain foods but not routinely monitored by public health authorities. DESIGN: To test the hypothesis, we treated wild-type mice, primary neuronal cultures, cell lines and isolated mitochondria with BMAA, and analysed its impact on gut microbiota composition, barrier permeability, inflammation and aSyn aggregation as well as in brain inflammation, dopaminergic neuronal loss and motor behaviour. To further examine the key role of mitochondria, we also determined the specific effects of BMAA on mitochondrial function and on inflammasome activation. RESULTS: BMAA induced extensive depletion of segmented filamentous bacteria (SFB) that regulate gut immunity, thus triggering gut dysbiosis, immune cell migration, increased intestinal inflammation, loss of barrier integrity and caudo-rostral progression of aSyn. Additionally, BMAA induced in vitro and in vivo mitochondrial dysfunction with cardiolipin exposure and consequent activation of neuronal innate immunity. These events primed neuroinflammation, dopaminergic neuronal loss and motor deficits. CONCLUSION: Taken together, our results demonstrate that chronic exposure to dietary BMAA can trigger a chain of events that recapitulate the evolution of the PD pathology from the gut to the brain, which is consistent with 'gut-first' PD.

Iron Supplementation and Exercise During Pregnancy: Effects on Behavior and the Dopaminergic System.

Although it is known that regular physical activity is recommended as part of a healthy lifestyle, the number of data concerning efficacy of exercise and your relationship with a demand for iron during pregnancy is limited. The purpose of this study was to evaluate the relationship between iron supplementation and exercise during pregnancy on the behavior of rats. Molecular variables dopamine transporter (DAT) and dopamine receptor (D2) related to the locomotor behavior in response to the exercise and the iron supplemented diet were investigated. Sixty-day-old female Wistar rats were used. The pregnant rats were distributed into the following groups: standard diet (SD, n = 7), iron supplementation (IS, n = 9), exercise (EX, n = 10), and exercise + iron supplementation (EX + IS, n = 9). All rats in both the pregnant and non-pregnant groups were submitted to open-field tests. The iron supplementation diet was shown to reduce locomotor behaviors, with reduced central and peripheral ambulation, reduced rearing, and increased freezing. On the other hand, physical exercise caused an increase in central and peripheral ambulation, and in rearing. The expression of the D2 receptor protein and the dopamine transporter DAT did not show changes with the interventions over 21 days of pregnancy. In this context, the present study demonstrated that both iron supplementation and exercise exerted an influence during pregnancy on the behavior of rats, however, with different effects.

Understanding and Improving the Oil and Water Barrier Performance of a Waterborne Coating on Paperboard.

Using paperboard as packaging material is more sustainable than using plastic. To be a viable replacement, however, the barrier properties of paperboard need to be improved. Applying a waterborne barrier coating for both oil and water is an attractive method to improve the barrier performance of paperboard food packaging. However, not much is known about the oil and water barrier properties and penetration pathways of such coatings. Here, an alkali-soluble resin (ASR)-stabilized waterborne emulsion polymer was prepared and applied on untreated paperboard. Its performance as oil and water barrier coating was investigated, and the penetration pathways for both oil and water through the coating are discussed. The presence of surface defects in the coating applied on the paperboard strongly affects both the oil and water barrier properties, but the coating's morphology and chemical nature only play a major role in the water barrier performance. The optimal barrier performance for oil and water was achieved when adding 5 wt % isopropanol (IPA) to the dispersion and applying two coating layers on paperboard. The IPA improves film formation and reduces the number of surface defects, which is explained by a more favorable spreading coefficient of the coating over the paperboard substrate. These insights will help to improve the oil and water barrier properties of polymer-coated paperboard for more sustainable packaging applications.

Assembly of partially covered strawberry supracolloids in dilute and concentrate aqueous dispersions.

HYPOTHESIS: Core-corona supracolloids can be assembled in aqueous dispersions by controlling the physical interactions between the corona and core colloidal particles. A raspberry corona configuration with full surface coverage of the core can be reached by inducing strong attractive interactions between the individual particles. A controlled partial surface coverage of the core, i.e. strawberry configuration, is however, more difficult to achieve. Supracolloids with different surface coverage ratio exhibit unique and multifunctional surface properties. EXPERIMENTS: By counterbalancing the multiple physical interactions playing a role during the assembly, the configuration and stability of the assemblies could be fine-tuned over a wide range of concentrations. Supracolloids consisting of polyethylene glycol (PEO)-grafted polymer particles covered by silica nanoparticles were assembled with different configurations, by adjusting the pH and ionic strength of the dispersion, the PEO grafting density and the particles concentration. The self-assembly process and resulting configurations were monitored via cryogenic transmission electron microscopy (Cryo-TEM) and light scattering. FINDINGS: The suitable conditions to assemble supracolloids with partial corona coverage have been established. Stable strawberry supracolloids could be prepared, both for diluted (1 wt%) and concentrated (12 wt%) dispersions. These hybrid supracolloids with well-defined configuration are highly relevant to developing advanced water-borne paints and inks, food dispersions, cosmetic and healthcare products.

Distribution of block copolymers in drying polymer films.

HYPOTHESIS: Block copolymers (BCP) consisting of a polar block and a surface active apolar block are widely used for surface functionalization of polymer films. The characteristics of the copolymer blocks determine whether surface segregation and/or phase separation occurs, for a given bulk mixture. This data can be used to find the optimal BCP composition where high surface enrichment is obtained without accumulation of phase separated BCP in the bulk. METHODS: The distribution of poly(ethylene oxide)-polydimethylsiloxane (PEO-PDMS) BCP in a polymer formulation relevant for coating applications is systematically investigated. The surface segregation is studied in liquid formulations with surface tension measurements and dried films with X-ray photoelectron spectroscopy (XPS), whereas phase separation is quantified using turbidity measurements. The results are compared with Scheutjens-Fleer self-consistent field (SF-SCF) computations, which are also applied to determine the effect of film drying on BCP phase stability and surface segregation. FINDINGS: Longer PDMS blocks result in lower interfacial tension of the liquid polymer mixture, whereas for the cured films, the largest PDMS concentration at the interface was obtained for intermediate PDMS block lengths. This is explained by the observation that phase separation already occurs at very low BCP concentrations for long PDMS blocks. The SCF predictions qualitatively agree with the experimental results and reveal that the BCP distribution changes significantly during film drying.

Anabolic steroids and their effects of on neuronal density in cortical areas and hippocampus of mice / Esteroides anabolizantes e seus efeitos na densidade neuronal em áreas corticais e no hipocampo de camundongos

Abstract Anabolic substances have been increasingly used by bodybuilders and athletes with the goal of improving performance and aesthetics. However, this practice has caused some concern to physicians and researchers because of unknowledge of consequences that the indiscriminate and illicit use of these substances can cause. Thus, this study analyzed the effects of two commercially available anabolic steroids (AS), Winstrol Depot® (Stanozolol) and Deposteron® (Testosterone Cypionate), in the neuronal density of limbic, motor and sensory regions on the cerebral cortex and in CA1, CA2, CA3 regions of the hippocampus. A total of 60 Swiss mice were used (30 males and 30 females), separated into three groups: control and two experimental groups, which received the AAS. From each brain, homotypic and semi-serial samples were taken in frontal sections from areas established for the study. The results showed that females treated with testosterone cypionate presented a reduction in all regions tested and the ones treated with Stanozolol showed a decrease in some hippocampal areas. Regarding male animals, stanozolol led to a decrease in neuron number in one hippocampal region. These data allow us to conclude that supra-physiological doses of steroids used in this study, can cause considerable damage to nervous tissue with ultrastructural and consequently behavioral impairment. These changes could interfere with the loss of physical yield and performance of athletes and non-athletes and may cause irreparable damage to individuals making irresponsible use of anabolic steroids.

Resumo As substancias anabólicas tem sido cada vez mais utilizadas por fisiculturistas e atletas com o objetivo de melhorar o desempenho e a estética. No entanto, essa prática tem causado algumas preocupações aos médicos e pesquisadores, devido ao desconhecimento das consequencias que o uso indiscriminado e ilícito dessas substâncias podem causar. Diante disso, este estudo analisou os efeitos de dois esteroides anabolizantes (EA) comercialmente disponíveis, Winstrol Depot® (Stanozolol) e Deposteron® (cipionato de testosterona), na densidade neuronal das regiões corticais límbica, motora e sensitive bem como das áreas CA1, CA2, CA3 do hipocampo. Foram utilizados 60 camundongos Swiss (30 machos e 30 fêmeas), separados em três grupos: controle e dois grupos experimentais, que receberam o EA. De cada cérebro, foram coletadas amostras homotípicas e semi-seriadas em cortes frontais das áreas estabelecidas para o estudo. Os resultados mostraram que as fêmeas tratadas com cipionato de testosterona apresentaram uma redução em todas as regiões analisadas já as fêmeas tratadas com Stanozolol mostraram uma diminuição em algumas áreas do hipocampo. Em relação aos animais machos, o stanozolol levou a uma diminuição na densidade neuronal em uma região do hipocampo. Estes dados nos permitem concluir que doses supra fisiológicas de esteroides utilizadas neste estudo podem causar danos consideráveis ao tecido nervoso com comprometimento ultraestrutural e consequentemente comportamental. Essas alterações podem interferir na perda de rendimento físico e no desempenho de atletas e não atletas e podem causar danos irreparáveis a indivíduos que fazem uso irresponsável destes EA.

From a eutectic mixture to a deep eutectic system via anion selection: Glutaric acid + tetraethylammonium halides.

In pursuit of understanding structure-property relationships for the melting point depression of binary eutectic mixtures, the influence of the anion on the solid-liquid (S-L) phase behavior was explored for mixtures of glutaric acid + tetraethylammonium chloride, bromide, and iodide. A detailed experimental evaluation of the S-L phase behavior revealed that the eutectic point is shifted toward lower temperatures and higher salt contents upon decreasing the ionic radius. The salt fusion properties were experimentally inaccessible owing to thermal decomposition. The data were inter- and extrapolated using various models for the Gibbs energy of mixing fitted to the glutaric-acid rich side only, which allowed for the assessment of the eutectic point. Fitting the experimental data to a two-parameter Redlich-Kister expansion with Flory entropy, the eutectic depth could be related to the ionic radius of the anion. The anion type, and in particular its size, can therefore be viewed as an important design parameter for the liquid window of other acid and salt-based deep eutectic solvents/systems.

Solvent Selectivity Governs the Emergence of Temperature Responsiveness in Block Copolymer Self-Assembly.

In highly selective solvents, block copolymers (BCPs) form association colloids, while in solvents with poor selectivity, they exhibit a temperature-controlled (de)mixing behavior. Herein, it is shown that a temperature-responsive self-assembly behavior emerges in solvent mixtures of intermediate selectivity. A biocompatible poly-ethylene(oxide)-block-poly-ε-caprolactone (PEO-PCL) BCP is used as a model system. The polymer is dissolved in solvent mixtures containing water (a strongly selective solvent for PEO) and ethanol (a poorly selective solvent for PEO) to tune the solvency conditions. Using synchrotron X-ray scattering, cryogenic transmission electron microscopy, and scanning probe microscopy, it is shown that a rich temperature-responsive behavior can be achieved in certain solvent mixtures. Crystallization of the PCL block enriches the phase behavior of the BCP by promoting sphere-to-cylinder morphology transitions at low temperatures. Increasing the water fraction in the solvent causes a suppression of the sphere-to-cylinder morphology transition. These results open up the possibility to induce temperature-responsive properties on demand in a wide range of BCP systems.

Photodynamic inactivation of Lasiodiplodia theobromae: lighting the way towards an environmentally friendly phytosanitary treatment.

The fungus Lasiodiplodia theobromae is one of the main causal agents of trunk canker and dieback of grapevine. The objective of this work was to evaluate the efficiency of photodynamic inactivation (PDI) of L. theobromae with synthetic and natural photosensitizers and irradiation with either sunlight or artificial photosynthetically active radiation. Although the growth of the mycelium could not be completely prevented with natural sunlight irradiation, phenothiazine dyes (methylene blue, MB; toluidine blue O, TBO), riboflavin and a cationic porphyrin (Tetra-Py+-Me) caused complete inhibition under continuous irradiation with artificial light. Free radicals were the main cytotoxic agents in the PDI with MB, indicating the predominance of the type I mechanism. PDI with MB or Tetra-Py+-Me may represent a promising approach for the sanitation of vine material in greenhouse nurseries, in order to reduce the risk of infection upon grafting.

Effects of age, weight, and housing system on prevalence of dead on arrival and carcass condemnation causes in laying hens.

Causes of condemnation and dead on arrival (DoA) birds were studied at the slaughterhouse and effects of age, BW, and housing system were investigated. A total of 1,156 (0.18%) birds were found DoA and 20,754 carcasses out of 629,331 (3.30%) were condemned during postmortem inspection. The main reasons for condemnation were peritonitis, septicemia, salpingitis, emaciation, and tumors. The mean percentages of condemnation by septicemia and emaciation differed according to age and BW groups and were more common in batches of younger (≤87 wk) and lighter (≤1.88 kg) birds. Moreover, peritonitis and ascites differed significantly between age groups, occurring more frequently in batches of older hens. The presence of tumors of the reproductive system was more frequently observed in older and heavier hens. This result raises the possibility of tumors being correlated with the higher number of prior ovulatory events. The total condemnation rate was lower in hens from organic systems, followed by free-range, and differed significantly from barns and cage systems. Carcasses with ascites and peritonitis were found more commonly in hens from cages and barns and both differed significantly from organic systems. Salpingitis was statistically more prevalent in barns, presenting differences from organic and free-range systems. Monitoring condemnation causes of end-of-lay hens at slaughter provided a better understanding of health and welfare issues in different housing systems and allowed to identify potential welfare problems, which can be used to improve management and welfare on farms.

Impact of Age, Education Level and Occupation on the Ability to Use the ICIQ-bladder Diary: Results From A Community Cohort Study Using a Predefined Clinical History.

OBJECTIVE: To assess difficulties in filling the International Consultation on Incontinence Questionnaire Bladder Diary (ICIQ-BD), determining the most troublesome items and patients' characteristics (age, education level, and professional activity) related to these difficulties. METHODS: A text composed of a clinical history of a fictitious patient with Lower Urinary Tract Symptoms (LUTS) was designed. 95 healthy volunteers (49 females, 46 males, mean age 45.6 ± 17.3 years) were asked to fill in the ICIQ-BD according to the data from the fictitious clinical history. Filling errors were analyzed, and patients' demographic characteristics tested as predictors. RESULTS: Bladder Sensation Score is the most problematic item of the ICIQ-BD, with only 38% of patients accurately filling this item. No association was found between total number of errors and age or education level, but participants working in less specialized professions had more difficulty in filling the BD (mean: 7.00 vs 3.94 errors, P= .002). CONCLUSION: ICIQ-BD is a reliable tool to assess for LUTS, but filling the Bladder Sensation Score item was more likely to be incorrectly filled by manual workers and volunteers with less intellectually demanding current professions.

Clinical decision-making and importance of the AAE/AAOMR position statement for CBCT examination in endodontic cases.

AIM: To compare conventional radiographic and cone beam computed tomography (CBCT) findings with reference to the American Association of Endodontics and American Academy of Oral and Maxillofacial Radiology (AAE/AAOMR) joint position statement and to determine the effect of the CBCT on the initial diagnoses and treatment plans in a single-centre Postgraduate Endodontic Programme. METHODOLOGY: The clinical CBCT scans of patients, treated at the Endodontic Department of the University of British Columbia, were reviewed for CBCT referrals by comparing them with corresponding radiographs. The features considered were periapical lesions, missed/extra canals, root fractures, complex anatomy, calcified canals and root resorption of tooth/teeth in question. Reasons for the CBCT prescriptions were assigned to 3 groups: to formulate the primary diagnosis, to confirm the diagnosis achieved by clinical examination and conventional radiographs, and to assist in treatment planning. Variables were compared statistically using chi-square and McNemar tests. RESULTS: A total of 128 CBCT examinations were performed on 110 patients. No CBCT examination was performed more than once on the same tooth. Overall, 76% of CBCT examinations were performed on previously root filled teeth. CBCT images revealed a significantly higher incidence of periapical lesions (P = 0.002), missed canals (P < 0.001), vertical root fractures (P = 0.004) and complex anatomy (P = 0.008) than periapical radiographs. CBCT was prescribed most frequently to assist surgical treatment planning (62%) rather than for generating a diagnosis (9%) or confirming diagnoses (29%). Both the diagnosis (P = 0.001) and the treatment plan (P = 0.005) initially made by examining periapical radiographs were altered significantly by the subsequent CBCT examination by revealing information such as new periapical lesions, missed canals or involvement of buccal or lingual cortical bone. CONCLUSION: CBCT examinations were prescribed mainly to assist treatment planning rather than for diagnosis. The majority of CBCT examinations were performed on previously root filled teeth. The additional information obtained from CBCT scans resulted in the alteration of the initial diagnoses as well as subsequent treatment plans in 59 out of 128 cases.

Anabolic steroids and their effects of on neuronal density in cortical areas and hippocampus of mice.

Anabolic substances have been increasingly used by bodybuilders and athletes with the goal of improving performance and aesthetics. However, this practice has caused some concern to physicians and researchers because of unknowledge of consequences that the indiscriminate and illicit use of these substances can cause. Thus, this study analyzed the effects of two commercially available anabolic steroids (AS), Winstrol Depot® (Stanozolol) and Deposteron® (Testosterone Cypionate), in the neuronal density of limbic, motor and sensory regions on the cerebral cortex and in CA1, CA2, CA3 regions of the hippocampus. A total of 60 Swiss mice were used (30 males and 30 females), separated into three groups: control and two experimental groups, which received the AAS. From each brain, homotypic and semi-serial samples were taken in frontal sections from areas established for the study. The results showed that females treated with testosterone cypionate presented a reduction in all regions tested and the ones treated with Stanozolol showed a decrease in some hippocampal areas. Regarding male animals, stanozolol led to a decrease in neuron number in one hippocampal region. These data allow us to conclude that supra-physiological doses of steroids used in this study, can cause considerable damage to nervous tissue with ultrastructural and consequently behavioral impairment. These changes could interfere with the loss of physical yield and performance of athletes and non-athletes and may cause irreparable damage to individuals making irresponsible use of anabolic steroids.

Microbial BMAA elicits mitochondrial dysfunction, innate immunity activation, and Alzheimer's disease features in cortical neurons.

BACKGROUND: After decades of research recognizing it as a complex multifactorial disorder, sporadic Alzheimer's disease (sAD) still has no known etiology. Adding to the myriad of different pathways involved, bacterial neurotoxins are assuming greater importance in the etiology and/or progression of sAD. ß-N-Methylamino-L-alanine (BMAA), a neurotoxin produced by some microorganisms namely cyanobacteria, was previously detected in the brains of AD patients. Indeed, the consumption of BMAA-enriched foods has been proposed to induce amyotrophic lateral sclerosis-parkinsonism-dementia complex (ALS-PDC), which implicated this microbial metabolite in neurodegeneration mechanisms. METHODS: Freshly isolated mitochondria from C57BL/6 mice were treated with BMAA and O2 consumption rates were determined. O2 consumption and glycolysis rates were also measured in mouse primary cortical neuronal cultures. Further, mitochondrial membrane potential and ROS production were evaluated by fluorimetry and the integrity of mitochondrial network was examined by immunofluorescence. Finally, the ability of BMAA to activate neuronal innate immunity was quantified by addressing TLRs (Toll-like receptors) expression, p65 NF-κB translocation into the nucleus, increased expression of NLRP3 (Nod-like receptor 3), and pro-IL-1ß. Caspase-1 activity was evaluated using a colorimetric substrate and mature IL-1ß levels were also determined by ELISA. RESULTS: Treatment with BMAA reduced O2 consumption rates in both isolated mitochondria and in primary cortical cultures, with additional reduced glycolytic rates, decrease mitochondrial potential and increased ROS production. The mitochondrial network was found to be fragmented, which resulted in cardiolipin exposure that stimulated inflammasome NLRP3, reinforced by decreased mitochondrial turnover, as indicated by increased p62 levels. BMAA treatment also activated neuronal extracellular TLR4 and intracellular TLR3, inducing p65 NF-κB translocation into the nucleus and activating the transcription of NLRP3 and pro-IL-1ß. Increased caspase-1 activity resulted in elevated levels of mature IL-1ß. These alterations in mitochondrial metabolism and inflammation increased Tau phosphorylation and Aß peptides production, two hallmarks of AD. CONCLUSIONS: Here we propose a unifying mechanism for AD neurodegeneration in which a microbial toxin can induce mitochondrial dysfunction and activate neuronal innate immunity, which ultimately results in Tau and Aß pathology. Our data show that neurons, alone, can mount inflammatory responses, a role previously attributed exclusively to glial cells.

Differential protein expression in diverse brain areas of Parkinson's and Alzheimer's disease patients.

Many hypotheses have been postulated to define the etiology of sporadic Parkinson's and Alzheimer's disorders (PD and AD) but there is no consensus on what causes these devastating age-related diseases. Braak staging of both pathologies helped researchers to better understand the progression and to identify their prodromal and symptomatic phases. Indeed, it is well accepted that Lewy body pathology and neurofibrillary tangles appearance correlates with disease progression and severity of symptoms in PD and AD, respectively. Additionally, several studies in PD and AD models try to disclose which cellular mechanisms are defaulted and trigger the neurodegenerative process that culminates with neuronal death causing PD and AD classical symptomatology. Herein, we determined expression levels of proteins involved in microtubule assembly, autophagic-lysosomal pathway and unfolded protein response in the cortex, hippocampus and SNpc of PD and AD patients, vascular dementia patients and aged-match controls. The differential expression allowed us to determine which pathways are determinant to synaptic dysfunction and to establish a time line for disease progression. Our results allow us to challenge the hypothesis that both PD and AD pathologies are caused by α-synuclein or Aß pathology propagation throughout the brain in a prion-like manner.