The anti-Parkinsonian drug selegiline delays the nucleation phase of α-synuclein aggregation leading to the formation of nontoxic species

Parkinson's disease (PD) is a movement disorder characterized by the loss of dopaminergic neurons in the substantia nigra and the formation of intraneuronal inclusions called Lewy bodies, which are composed mainly of α-synuclein (α-syn). Selegiline (Sel) is a noncompetitive monoamino oxidase B inhibitor that has neuroprotective effects and has been administered to PD patients as monotherapy or in combination with l-dopa. Besides its known effect of increasing the level of dopamine (DA) by monoamino oxidase B inhibition, Sel induces other effects that contribute to its action against PD. We evaluated the effects of Sel on the in vitro aggregation of A30P and wild-type α-syn. Sel delays fibril formation by extending the lag phase of aggregation. In the presence of Sel, electron microscopy reveals amorphous heterogeneous aggregates, including large annular species, which are innocuous to a primary culture enriched in dopaminergic neurons, while their age-matched counterparts are toxic. The inhibitory effect displayed by Sel is abolished when seeds (small fibril pieces) are added to the aggregation reaction, reinforcing the hypothesis that Sel interferes with early nuclei formation and, to a lesser extent, with fibril elongation. NMR experiments indicate that Sel does not interact with monomeric α-syn. Interestingly, when added in combination with DA (which favors the formation of toxic protofibrils), Sel overrides the inhibitory effect of DA and favors fibrillation. Additionally, Sel blocks the formation of smaller toxic aggregates by perturbing DA-dependent fibril disaggregation. These effects might be beneficial for PD patients, since the sequestration of protofibrils into fibrils or the inhibition of fibril dissociation could alleviate the toxic effects of protofibrils on dopaminergic neurons. In nondopaminergic neurons, Sel might slow the fibrillation, giving rise to the formation of large nontoxic aggregates

Polymorphisms of BoLA-DRB3 gene and its association with resistance / susceptibility to Leucosis in Holstein cattle from La Pampa

La leucosis bovina enzoótica es una enfermedad del ganado bovino adulto causada por el retrovirus de la leucemia bovina. Se puede manifestar como: una forma asintomática aleucémica, con un número normal de linfocitos B en sangre; una forma de linfocitosis permanente, con aumento del número de linfocitos B y como una presentación linfoproliferativa tumoral en forma de linfosarcoma. Los alelos de los genes del Complejo Principal de Histocompatibilidad Bovino (BoLA) han sido asociados con resistencia y susceptibilidad a enfermedades infecciosas. El objetivo general del presente estudio consistió en asociar los polimorfismos del exón 2 del gen BoLA-DRB3.2, definidos mediante la técnica de Reacción en Cadena de la Polimerasa y la técnica de Polimorfismos de la Longitud de los Fragmentos de Restricción (PCR-RFLP), con resistencia/susceptibilidad a leucosis en vacas Holstein de La Pampa. Se basó en un diseño caso/control, para lo cual se tomó muestra de sangre a 150 animales en 3 oportunidades con intervalos de tres meses cada una. Los animales fueron incluidos en el grupo caso cuando dieron positivos en la prueba de inmunodeficiencia en agar (DIDA) y cuyo recuento linfocitario en sangre fue ≥ 10.000 linfocitos/μl y el grupo control estuvo constituido con animales negativos en DIDA y que tenían < 10.000 linfocitos/μl de sangre. Los tests Exacto de Fisher y Odds Ratio (OR) de Woolf-Haldane se utilizaron para estudiar la asociación entre recuento de linfocitos y resultados del DIDA con las variantes alélicas. En 82 animales genotipados por PCRRFLP el alelo DRB3.2*22 evidenció un OR= 5,332 (p= 0,0138) y el alelo DRB3.2*11 mostró un OR= 0,11 (p=0,001) siendo más frecuentes en el grupo caso y en el grupo control, respectivamente. Estos resultados evidenciarían que vacas con el alelo DRB3.2*22 presentarían mayor riesgo a desarrollar leucosis y vacas con el alelo DRB3.2*11 un menor riesgo (resistencia). El análisis de la relación entre alelos del gen BoLA-DRB3.2 y resistencia/ susceptibilidad a leucosis podría mejorarse con investigaciones profundas en linajes familiares de vacas lecheras.EBL is a disease of adult cattle caused by the retrovirus, bovine leukemia. It can manifest: aleukemic an asymptomatic form, with a normal number of B lymphocytes in the blood; a form of permanent lymphocytosis, with an increase in the number of B lymphocytes and finally a lymphoproliferative tumor presentation in the form of lymphosarcoma. The alleles of the genes of the Bovine Major Histocompatibility Complex (BoLA) have been associated with resistance and susceptibility to infectious diseases. The overall objective of this study was to associate polymorphisms of the BoLA-DRB3.2 gene, defined by the technique of polymerase chain reaction technique and polymorphisms length of the restriction fragment (PCR-RFLP), with resistance / susceptibility to leucosis in Holstein cows of La Pampa. It relied on a case/control design, for which blood samples were taken to 150 animals in 3 opportunities with intervals of three months each. The case group comprised animals that were positive in the immunodiffusion agar animals test (DIDA) and whose blood lymphocyte count was ≥ 10.000 linf/μl and the control group included cows that ware negative for DIDA and present &lt; 10.000 linf/μl of blood. Fisher’s Exact test and Odds Ratio (OR) of Woolf-Haldane were used to study the association between lymphocyte count and DIDA results with allelic variants. In 82 animals genotyped by PCR-RFLP, the DRB3.2*22 allele showed an OR = 5.332 (p = 0.0138) and DRB3.2*11 allele had a OR value of 0.11 (p=0.001) and were the most frequent in the control group and in the case group, respectively. These results would showed DRB3.2*22 allele and DRB3.2*11 allele a high risk of having leucosis (susceptibility) and low risk to suffer (resistance) respectively. The analysis of the relationship between alleles of the BoLA and resistance/susceptibility to leucosis could be improved with deep research on family lineages of dairy cows.Instituto de Genética Veterinari

Dopamine affects the stability, hydration, and packing of protofibrils and fibrils of the wild type and variants of alpha-synuclein

Parkinson's disease (PD) is characterized by the presence of cytoplasmic inclusions composed of alpha-synuclein (alpha-syn) in dopaminergic neurons. This suggests a pivotal role of dopamine (DA) on PD development. Here, we show that DA modulates differently the stability of protofibrils (PF) and fibrils (F) composed of wild type or variants of alpha-syn (A30P and A53T) as probed by high hydrostatic pressure (HHP). While in the absence of DA, all alpha-syn PF exhibited identical stability, in its presence, the variant-composed PF acquired a greater stability (DAPFwt < DAPFA30P = DAPFA53T), implying that they would last longer, which could shed light onto why these mutations are so aggressive. When alpha-syn was incubated for long times (18 days) in the presence of DA, we observed the formation of F by electronic microscopy, suggesting that the PF trapped in the presence of DA in short times can evolve into F. The stability of F was also altered by DA. DAFwt was more labile than Fwt, indicating that the former would be more susceptible to breakage. PFA30P and DAPFA30P, when added to mesencephalic and cortical neurons in culture, decreased the number and length of neurites and increased the number of apoptotic cells. Surprisingly, these toxic effects of PFA30P and DAPFA30P were practically abolished with HHP treatment, which was able to break the PF into smaller aggregates, as seen by atomic force microscopy. These results suggest that strategies aimed at breaking and/or clearing these aggregates is promising in alleviating the symptoms of PD

Echinococcus granulosus Antigen B Structure: Subunit Composition and Oligomeric States

Antigen B (AgB) is the major secretory protein of the Echinococcus granulosus hydatid cyst, the causative agent of cystic hydatid disease. Structurally, AgB is a multisubunit protein formed by 8-kDa subunits, but it is not known which subunits are secreted by a single parasite (cyst) and how they interact in the formation of distinct AgB oligomeric states. Here, we investigated AgB subunit composition and oligomeric states in individual samples from bovine and human cysts. We identified AgB8/1, AgB8/2, AgB8/3 and AgB8/4 subunits in AgB oligomers of all samples analyzed. Quantitative and qualitative differences in the expression of AgB subunits were observed within and between samples. Using recombinant subunits as models, we showed that AgB subunits form distinct oligomeric states, with a rAgB8/3>rAgB8/2>rAgB8/1 maximum size relation. We also demonstrated by different experimental approaches that rAgB8/3 oligomers are more similar, both in size and morphology, to those observed for E. granulosus AgB. Overall, we provided experimental evidences that AgB is composed of different subunits within a single cyst, and that subunits have different abundances and oligomerization properties. These issues are important for the understanding of AgB expression and structure variations, and their impact for the host-parasite cross-talk

Aspectos físico-químicos e biológicos de ureases vegetais

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Aspectos físico-químicos e biológicos de ureases vegetais

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Fármacos multifuncionais: monoamina oxidase e &#945;-sinucleína como alvos terapêuticos na doença de Parkinson

Parkinson's disease (PD) is a neurodegenerative disorder associated to selective degeneration of dopaminergic neurons caused by an intricate relationship among dopamine metabolism, oxidative stress and &#945;-synuclein fibrillation. Most therapies for PD have focused on dopamine replacement through the use of both monoamine oxidase inhibitors (MAOIs) and dopamine precursor L-dopa. Interestingly, certain MAOIs have a broad spectrum of action including anti-fibrillogenic properties in &#945;-synuclein aggregation. Herein we revisit the chemical properties of MAOIs and their action on important targets associated with PD, notably &#945;-synuclein fibrillation and dopamine metabolism, discussing the strategies associated with the development of multi-target drugs for neurodegenerative diseases