Detección de anticuerpos contra Neospora caninum en caninos del área urbana y rural de Cumaral, Meta-Colombia

RESUMEN Objetivo. Estimar la seroprevalencia a Neospora caninum en caninos del área urbana y rural de Cumaral, Meta y determinar algunos factores de riesgo asociados a la seropositividad. Materiales y métodos. Se efectuó un estudio transversal en 222 perros (112 perros del área urbana y 110 del área rural). El tamaño de la muestra fue calculado en el programa Epidat v. 3.1. Los sueros sanguíneos fueron analizados mediante la técnica de Inmunofluorescencia Indirecta para IgG con un kit comercial. Los análisis de frecuencias, chi-cuadrado, fueron realizados mediante el paquete estadístico SPSS v. 25.0 Resultados. La seroprevalencia general fue 36.9% (IC95%: 30.9-43.5 %). La seropositividad entre los grupos fue: urbana (38.4 %) y rural (35.5%) (p>0.05), machos (36.9%) y hembras (36.9%) (p >0.05); en cachorros (32.7%), jóvenes (40.0%) y adultos (37.4%) (p>0.05), en contacto con predios pecuarios (40.7%) y sin contacto (35.2%) (p>0.05). Conclusiones. La seroprevalencia observada fue alta en las dos poblaciones analizadas y sugiere que los caninos han estado en contacto con el parásito, posiblemente por diferentes fuentes de infección que requieren ser estudiadas posteriormente.

ABSTRACT Objective. To estimate the seroprevalence to Neospora caninum in canines of the urban and rural area of Cumaral, Meta and determine some risk factors associated with seropositivity. Materials and methods. A cross-sectional study was carried out in 222 dogs (112 dogs from the urban area and 110 dogs from the rural area), the sample size was calculated by using Epidat v program. 3.1. The sera were analyzed using the Indirect Immunofluorescence technique for IgG with a commercial kit. Frequency analyzes by chi-square of independence were performed in SPSS v. 25.0 Results. The general seroprevalence was 36.9% (95% CI: 30.9-43.5%). The seropositivity between the groups was: urban (38.4%) and rural (35.5%) (p>0.05), males (36.9%) and females (36.9%) (p>0.05); in puppies (32.7%), youth (40.0%) and adults (37.4%) (p>0.05), in contact with livestock farms (40.7%) and without contact (35.2 %) (p>0.05), Conclusions. The seroprevalence observed was high in the two populations analyzed and suggests that the canines have been in contact with the parasite, possibly due to different sources of infection that need to be studied later.

PREVALENCIA DE Babesia caballi, Theileria equi Y TRIPANOSOMIASIS Y ANÁLISIS DE FACTORES DE RIESGO EN EQUINOS DE ANTIOQUIA, COLOMBIA / PREVALENCE OF Babesia caballi, Theileria equi AND TRYPANOSOMIASIS AND ANALYSIS OF RISK FACTORS IN HORSES OF ANTIOQUIA, COLOMBIA

RESUMEN Esta investigación, se llevó a cabo, debido a la poca información nacional existente en hemoparásitos equinos y su objetivo fue determinar la prevalencia de babesiosis y tripanosomiasis, en el Valle de Aburrá y Rionegro, municipios de Antioquia (Colombia), así como algunos factores de riesgo asociados a la presentación de seropositividad, a estas enfermedades. En 223 predios, con una población de 1.008 equinos, se tomó muestra de sangre venosa, para realizar el diagnóstico serológico y molecular. La información epidemiológica fue obtenida, a través de la aplicación de una encuesta dirigida a las personas a cargo de los animales. Para el análisis estadístico, se aplicó Chi cuadrado de independencia y la Prueba Exacta de Fisher, cuando fue necesario. Para las asociaciones bivariadas, se calcularon estimaciones del riesgo (OR) por variable explicativa, con sus respectivos intervalos de confianza, del 95% (I.C. 95%). Se encontró una prevalencia del 11,9%, para babesiosis y de 1,9%, para tripanosomiasis; como factor de protección, se encontró el hecho de salir a una feria. Para la piroplasmosis, el lugar geográfico, el sexo, estar castrado, ser positivos a Anemia Infecciosa Equina (AIE), estrongílidos u oxiurus fueron factores de riesgo. Para tripanosomiasis, el factor de riesgo fue estar infestado con Dermacentor nitens o ser mular. Se deben reforzar métodos de vigilancia epidemiológica activa, sobre todo, en casos de movilización continua de equinos o desarrollo de eventos, que involucren presencia masiva de ejemplares.

ABSTRACT This research was carried out due to the little existing national information on equine hemoparasites; the objective was to determine babesiosis and trypanosomiasis prevalence in the Aburrá Valley and Rionegro Antioquia (Colombia) as well as some risk factors associated with the seropositivitie of these diseases. In 223 lots with a population of 1.008 equine venous blood sample was taken for serological diagnosis and molecular. Epidemiological information was obtained through the application of a survey addressed to persons in charge of the animals. For the statistical analysis, Chi square of independence or Fisher's Exact Test was applied when necessary. For the bivariate associations, risk estimates (OR) were calculated by explanatory variable, with their respective 95% confidence intervals (95% CI).It was found a prevalence of 11.9% for babesiosis and 1.9% for trypanosomiasis and as protection factor found the fact of going to trade fairs. Geographic location, sex and being neutered, be positive for Equine Infectious Anemia (AIE), strongyles or oxiurus was a risk factor for piroplasmosis. For Trypanosomiasis the risk factor was to be infested with ticks Dermacentor nitens or be mule. Methods of active epidemiological surveillance should be reinforced, especially in cases of continuous mobilization of equines or development of events that involve massive presence of horses.

A preliminary survey of Trichinella spp. in pigs raised under controlled housing conditions in Colombia: 2014-2016.

A preliminary survey of Trichinella spp. infection was conducted in Colombian swine herds between 2014 and 2016. A total of 1,773 pigs reared on farms under controlled housing conditions and processed in 34 slaughterhouses were tested either by the artificial digestion of pooled muscle samples (n = 1,173) or by serology (n = 600). In addition, 550 rats trapped on 29 swine farm premises were also tested by artificial digestion. No positive pig samples were detected. Similarly, no Trichinella spp. muscle larvae were detected in rats. These results are in agreement with the lack of historical Trichinella infection reports in domestic and wild animals and humans in Colombia. However, a more extensive epidemiological investigation and a continuous surveillance program are needed to continue declaring swine herds in Colombia free of Trichinella infection.

Prevalencia de parásitos gastrointestinales en equinos y factores de riesgo asociados en varias zonas de Antioquia, Colombia / Prevalence of gastrointestinal parasites in equines and associated risk factors in several areas of Antioquia, Colombia / Prevalência de parasitos gastrointestinais em equinos e fatores de risco associados em várias zonas de Antioquia, Colômbia

Resumen Existen pocos reportes de la prevalencia de parásitos gastrointestinales en equinos en Colombia. En este estudio se muestrearon 946 caballos de un total de 204 predios en los municipios de Rionegro y los que comprenden el Valle de Aburrá departamento de Antioquia. La materia fecal fue tomada directamente del recto y procesada por las técnicas de McMaster׳ Baermann y Graham. La prevalencia de parásitos a nivel de predios fue del 563%׳ para Strongylidae׳ 7%׳10 para Strongyloides spp8%׳2 ׳ para Oxyuridae y 03%׳ para Dictyocaulus spp. Desde el punto de vista práctico׳ los más patogénicos y de mayor problema de resistencia son los Cyathostomidos׳ que se incluyen dentro de los Strongylidae y cuya prevalen-cia fue superior al 50%. El OR de presentar Strongylidae es 49% menor en equinos que participan en ferias en comparación con los que no lo hacen. El OR de presentar Strongylidae es 56% menor en equinos que usan pesebrera en comparación con los que no la usan. Por su parte׳ el OR de ser positivo a Oxyuris equi es 88% menor en animales que salen a ferias en comparación con los que no salen a feria y el OR de ser positivo a Oxyuris equi es 366׳ veces mayor en equinos con condición corporal <2 en comparación con equinos de condición corporal >3. Con base en esta información es importante iniciar estudios de resistencia a antihelmínticos y evaluar la relación entre la carga parasitaria y las posibles afecciones clínicas para así determinar el impacto real de estas parasitosis en los equinos de estas regiones.

Abstract There are few reports on the prevalence of gastrointestinal parasites in equines in Colombia. In this study946 ׳ horses were sampled from a total of 204 farms in the municipalities of Rionegro and those that comprise the Valle de Aburrá׳ in the department of Antioquia. Fecal samples were taken directly from the rectum and processed by the McMaster׳ Baermann and Graham techniques. The prevalence of parasites at the farm level was 56.3% for Strongyüdae10.7% ׳ for Strongyloides spp2.8% ׳ for Oxyuridae and 0.3% for Dictyocaulus spp. From a practical point of view׳ the most pathogenic parasites and of greatest resistance concern were the Cyathostomids׳ which are included within the Strongylidae and whose prevalence was greater than 50%. The OR of presenting Strongylidae was 49% lower in equines that participated in fairs compared to those that did not. The OR of presenting Strongylidae was 56% lower in equines that use stables than those not being stalled. A1so׳ the OR of being positive to Oxyuris equi was 88% lower in animals that participate in fairs compared to those that did not׳ and was 3.66 times higher in equines with body condition < 2 in comparison with those of body condition >3. Based on this information it is important to initiate studies that evaluate the state of antihelminthic resistance and the relationship between parasitic load and clinical condition in order to determine the real impact of these parasites on the equines of these regions.

Resumo Existem poucos reportes da prevalência de parasitos gastrointestinais em equinos na Colômbia. Nessa pesquisa foram amostrados 946 cavalos de um total de 204 fazendas nos municípios de Rionegro e dos que formam o Vale de Aburrá׳ no departamento de Antióquia. A matéria fecal foi tirada diretamente do reto e processada pelas técnicas de McMaster׳ Baermann e Graham. A prevalência de parasitos das fazendas foi de 56.3% para Strongylidae7%׳10 ׳ para Strongyloides spp8%׳2 ׳ para Oxyuridae y 0.3% para Dictyocaulus spp. Desde o ponto de vista prático׳ os mais patogênicos e de maior problema de resistência são os Cyathostomidos׳ que estão incluídos dentro dos Stronglylidae e cuja prevalência foi superior ao 50%. O OR de apresentar Strongylidae é 49% menor nos equinos que participam em férias comparado com os que não. O OR de apresentar Strongylidae é 56% menor nos equinos que usam manjedoura comparado com os que não a usam. Por outro lado׳ o OR de ser positivo a Oxyuris equi é 88% menor em animais que vão a férias comparado com os que não vão e o OR de ser positivo a Oxyuris equi é 3.66 vezes maior nos equinos com condição corporal <2 comparado com os equinos de condição corporal >3. Com base nesta informação׳ se conclui que é importante iniciar estudos de resistência a anti-helmínticos e avaliar a relação entre a carga parasitária e as possíveis afecções clínicas para assim determinar o impacto real destas parasitoses nos equinos destas regiões.

Cross-sectional study of 13 Leptospira serovars in cows in a Colombian dairy region / Estudio transversal de 13 serovares de Leptospira en vacas en una región lechera colombiana / Estudo transversal de 13 sorotipos de Leptospira em vacas em uma região leitera colombiana

Abstract Background: Leptospirosis in cattle is recognized as a major cause of reproductive loses due to abortions, early embryonic death, and infertility. The type of Leptospira serovars that are currently circulating in Colombian herds is largely unknown due to the lack of national reports. Objective: The prevalence of antibodies specific to 13 Leptospira spp serovars was investigated in unvaccinated cows from 26 herds in San Pedro de los Milagros (Province of Antioquia) dairy region. Methods: Microscopic agglutination test was used on serum samples, following WHO recommendations. Results: A total of 112/900 (12.4%) of the animals had antibody titers of 1:100 or greater to at least one Leptospira serovar. Of those positive, 34/900 (3,8%) animals seroconverted to two or more serovars. In order of decreasing prevalence, the serovars detected were: hardjo prajitno, pomona, grippotyphosa, tarassovi, copenhageni, canicola, cellodeni and bratislava. There were 28 animals seropositive to serovar hardjo. Conclusion: Our results indicate that leptospirosis should be included as a differential diagnosis for various reproductive problems in this region.

Resumen Antecedentes: La leptospirosis es la mayor causal de pérdidas reproductivas debidas a abortos, muerte embrionaria temprana e infertilidad. Se desconocen los serovares de Leptospira que actualmente se encuentran circulando en el rebaño colombiano debido a la falta de reportes nacionales. Objetivo: Se investigó la prevalencia de anticuerpos específicos para 13 serovares de Leptospira spp en vacas no vacunadas de 26 rebaños en la región lechera de San Pedro de los Milagros (Provincia de Antioquia). Métodos: La prueba de aglutinación microscópica fue empleada en muestras de suero de acuerdo a las recomendaciones de la OMS. Resultados: Un total de 112/900 (12,4%) de los animales presentaron títulos de anticuerpos de 1:100 o superiores para al menos un serovar de Leptospira. Entre los positivos, el 34/900 (3,8%) de los animales presentaron seroconversión para dos o más serovares. En orden decreciente de prevalencia, los serovares detectados fueron: hardjo prajitno, pomona, grippotyphosa, tarassovi, copenhageni, canicola, cellodeni y Bratislava. En total, 28 animales fueron seropositivos al serovar hardjo. Conclusión: La leptospirosis debería ser incluida como diagnóstico diferencial en problemas reproductivos en esta región.

Resumo Antecedentes: Aleptospirose em bovinos é reconhecida como uma das principais causas de perdas reprodutivas devido a abortos, morte embrionária precoce e infertidade. Os sorotipos de Leptospira que estão atualmente em circulação no rebanho colombiano não se conhecem devido à falta de reportes nacionais. Objetivo: Neste estúdio foi investigada a prevalência de anticorpos específicos para 13 sorovares de Leptospira spp em vacas não vacinadas de 26 rebanhos leiteiros da região de San Pedro, Antioquia. Métodos: Foi empregado o teste de aglutinação microscópica (MAT) em amostras de soro de acordo com as recomendações da OMS. Resultados: Um total de 112/900 (12,4%) animais apresentaram títulos de anticorpos de 1:100 ou superiores para pelo menos um sorotipo de Leptospira. Dos animais positivos, 34/900 (3,8%) soroconverteram para dois ou mais sorotipos. A fim de diminuir a prevalência, foram detectados os sorotipos: hardjo, prajitno, pomona, grippotyphosa, tarassovi, copenhageni, canicola, cellodeni e Bratislava. Um total de28 animais foram soropositivos para o sorotipo hardjo. Conclusão: Os resultados indicam que a leptospirose deveria ser incluída nos diagnósticos diferenciais para o diagnóstico diferencial de problemas em doenças reprodutivas nos bovinos de Antioquia.

Regional report on Angiostrongylus vasorum in Colombia: Genetic similarity to European lineage.

The canine lungworm Angiostrongylus vasorum is considered neglected in South America and was only sporadically reported in dogs and wildlife. Gastropods act as obligatory intermediate hosts for this parasitosis. We here analysed Achatina fulica (African giant snail) populations from 5 regions of Colombia for A. vasorum infections. In total, 609 snails were collected from the departments Antioquia, Valle del Cauca and Putumayo. Angiostrongylus vasorum-infected A. fulica were found in all departments with a total prevalence of 3.9%. Larvae originating from Putumayo were molecularly characterized and identified as the European lineage of A. vasorum. This regional report shows for the first time the presence of A. vasorum in intermediate hosts in Colombia and the European genotype in South America.

Molecular characterization of Marek's disease virus in a poultry layer farm from Colombia.

Marek's disease (MD) is a lymphoproliferative disease caused by an Alphaherpesvirus, genus Mardivirus, serotype 1 (Gallid Herpesvirus 2, GaHV-2) that includes all known pathogenic strains. In addition to Marek's disease virus (MDV) serotype 1, the genus includes 2 distinct nonpathogenic serotypes: serotype 2 (GaHV-3) and serotype 3 (Meleagridis Herpesvirus 1, MeHV-1) which are used in commercially available vaccines against MD. As a result of vaccination, clinical signs are not commonly observed, and new cases are usually associated with emerging variant strains against which the vaccines are less effective. In this study, a commercial layer farm showing clinical signs compatible with MDV infection was evaluated. Histological lesions and positive immunohistochemistry in the sciatic nerve and thymus were compatible with cytolytic phase of MD. GaHV-2, GaHV-3 and MeHV-1 were identified by PCR and qPCR in blood samples from 17 birds with suspected MD. Analysis of the Meq gene of the Colombian GaHV-2 isolate revealed a 99% sequence identity with Asian strains, and in the phylogenetic analysis clustered with vv+ MDV. The analysis of amino acid alignments demonstrated an interruption of the proline rich region in P176A, P217A and P233L positions, which are generally associated with vv+ strains. Some of these changes, such as P233L and L258S positions have not been reported previously. In addition, primary cell cultures inoculated with lymphocytes isolated from the spleen showed typical cytopathic effect of GaHV-2 at 5 d post infection. Based on the molecular analysis, the results from this study indicate the presence of vv+ MDV infection in commercial birds for the first time in Colombia. It is recommended to perform further assays in order to demonstrate the pathotype characteristics in vivo.

In vitro resistance to topical acaricides of the cattle tick Rhipicephalus (Boophilus) microplus from four regions of Colombia / Resistencia in vitro a acaricidas tópicos de poblaciones de garrapatas Rhipicephalus (Boophilus) microplus provenientes de cuatro departamentos de Colombia / Resistência in vitro aos acaricidas tópicos de populações de carrapatos Rhipicephalus (Boophilus) microplus provenientes de quatro departamentos da Colô mbia

In Colombia, the control of the cattle tick Rhipicephalus (Boophilus) microplus lies almost exclusively on the use of chemical acaricides. However, R. (B.) microplus has developed resistance to most of the commercially available products. The aim of this study was to assess the degree of resistance to the major topical acaricides used in Colombia. The adult immersion test was used as recommended by FAO with ticks from 7 farms originating in 4 distant Colombian regions. The efficacy of the products was calculated based on the reproductive parameters of fecundity and fertility with groups of ticks (n = 40) exposed to the recommended concentrations of each acaricide, and compared with the respective control group exposed to distilled water. In general, products containing deltamethrin (25 ppm) and amitraz (208 ppm) were less effective, with efficacy values of <20% and 10-50%, respectively. Doubling the concentration did not increase the efficacy of either compound. The combination of chlorpyrifos (312 ppm) + cypermethrin (150 ppm) showed great variation between farms, with an efficacy ranging between 22-93%. Unlike chlorpyrifos, the organophosphorus ethion (622 ppm) was 99-100% effective in completely inhibiting oviposition in 4 of the 7 farms. In conclusion, resistance to deltamethrin and amitraz is very high in seven distant strains of the Colombian territory, varies widely for chlorpyrifos, and was not detected for ethion that was 99-100% effective

El control de la garrapata Rhipicephalus (Boophilus) microplus en Colombia radica casi exclusivamente en el uso de acaricidas químicos. Sin embargo, R. (B.) microplus ha desarrollado resistencia a la mayoría de los productos actualmente disponibles comercialmente. El objetivo de este estudio fue valorar el grado de resistencia a concentraciones recomendadas de los principales acaricidas comerciales de uso tópico en Colombia. Se empleó la prueba de inmersión de adultas (teleoginas) recomendada por la FAO con garrapatas procedentes de siete predios localizados en cuatro departamentos colombianos. La eficacia de los productos se calculó con base en los parámetros reproductivos de fecundidad y fertilidad con grupos de garrapatas (n=40) expuestas a cada acaricida, comparándolos con el respectivo grupo control expuesto a agua destilada. En general, los productos de deltametrina (25 ppm) y amitraz (208 ppm) fueron los menos efectivos, con valores de eficacia de <20% y de entre 10-50%, respectivamente. Tampoco se observó un aumento de eficacia al doblar la concentración de ambos compuestos por separado. La combinación de clorpirifos (312 ppm) + cipermetrina (150 ppm) mostró gran variación entre fincas, con una eficacia de entre el 22-93%. A diferencia  del clorpirifos, el organofosforado etión (622 ppm) sí produjo un 99-100% de eficacia inhibiendo completamente la oviposición en cuatro de las siete fincas. En conclusión, la resistencia a deltametrina y amitraz es muy alta en siete poblaciones distantes del territorio colombiano; en cuanto al clorpirifos varía mucho, y éste no parece tener resistencia cruzada con el etión que fue el de mayor eficacia.

O controle do carrapato Rhipicephalus (Boophilus) microplus na Colô mbia está focado quase exclusivamente no uso de acaricidas químicos. Porém, R. (B.) microplus tem desenvolvido resistência à maioria dos produtos disponíveis comercialmente. O objetivo deste estudo foi valorar o grau de resistência às concentrações recomendadas dos principais acaricidas comerciais de uso tópico na Colô mbia. Foi usada a prova da imersã o de adultas (teleoginas) recomendada pela FAO com carrapatos procedentes de 7 fazendas localizadas em 4 departamentos colombianos. A eficácia dos produtos calculou-se com base nos parâmetros reprodutivos de fecundidade e fertilidade com grupos de carrapatos (n=40) expostos a cada acaricida. Em geral, os produtos deltametrina (25 ppm) e amitraz (208 ppm) foram os menos efetivos, com valores de eficácia de  <20% e de entre 10-50%, respectivamente. Também nã o foi observado aumento da eficácia ao dobrar a concentraçã o de ambos produtos por separado. A combinaçã o de clorpirifos (312 ppm) + cipermetrina (150 ppm) mostrou uma grande variaçã o entre as fazendas, com uma eficácia de 22-93%. Diferentemente do clorpirifos, o organofosforado etion (622 ppm) teve um 99-100% de eficácia por inibiçã o completa da ovoposiçã o em 4 das 7 fazendas. Em conclusã o, a resistência à deltametrina e amitraz é muito alta em 7 populações distantes do território colombiano; quanto ao clorpirifos, varia muito e este nã o parece ter resistência cruzada com o etion que foi o de maior eficácia.<20% e de entre 10-50%, respectivamente. Também não foi observado aumento da eficácia ao dobrar a concentração de ambos produtos por separado. A combinação de clorpirifos (312 ppm) + cipermetrina (150 ppm) mostrou uma grande variação entre as fazendas, com uma eficácia de 22-93%. Diferentemente do clorpirifos, o organofosforado etion (622 ppm) teve um 99-100% de eficácia por inibição completa da ovoposição em 4 das 7 fazendas. Em conclusão, a resistência à deltametrina e amitraz é muito alta em 7 populações distantes do território colombiano; quanto ao clorpirifos, varia muito e este não parece ter resistência cruzada com o etion que foi o de maior eficácia.

BACE1 RNAi Restores the Composition of Phosphatidylethanolamine-Derivates Related to Memory Improvement in Aged 3xTg-AD Mice.

ß-amyloid (Aß) is produced by the ß-secretase 1 (BACE1)-mediated enzymatic cleavage of the amyloid precursor protein through the amyloidogenic pathway, making BACE1 a therapeutic target against Alzheimer's disease (AD). Alterations in lipid metabolism are a risk factor for AD by an unknown mechanism. The objective of this study was to determine the effect of RNA interference against BACE1 (shBACEmiR) on the phospholipid profile in hippocampal CA1 area in aged 3xTg-AD mice after 6 and 12 months of treatment compared to aged PS1KI mice. The shBACEmiR treatment induced cognitive function recovery and restored mainly the fatty acid composition of lysophosphatidylethanolamine and etherphosphatidylethanolamine, reduced the cPLA2's phosphorylation, down-regulated the levels of arachidonic acid and COX2 in the hippocampi of 3xTg-AD mice. Together, our findings suggest, for the first time, that BACE1 silencing restores phospholipids composition which could favor the recovery of cellular homeostasis and cognitive function in the hippocampus of triple transgenic AD mice.

p35 and Rac1 underlie the neuroprotection and cognitive improvement induced by CDK5 silencing.

CDK5 plays an important role in neurotransmission and synaptic plasticity in the normal function of the adult brain, and dysregulation can lead to Tau hyperphosphorylation and cognitive impairment. In a previous study, we demonstrated that RNAi knock down of CDK5 reduced the formation of neurofibrillary tangles (NFT) and prevented neuronal loss in triple transgenic Alzheimer's mice. Here, we report that CDK5 RNAi protected against glutamate-mediated excitotoxicity using primary hippocampal neurons transduced with adeno-associated virus 2.5 viral vector eGFP-tagged scrambled or CDK5 shRNA-miR during 12 days. Protection was dependent on a concomitant increase in p35 and was reversed using p35 RNAi, which affected the down-stream Rho GTPase activity. Furthermore, p35 over-expression and constitutively active Rac1 mimicked CDK5 silencing-induced neuroprotection. In addition, 3xTg-Alzheimer's disease mice (24 months old) were injected in the hippocampus with scrambled or CDK5 shRNA-miR, and spatial learning and memory were performed 3 weeks post-injection using 'Morris' water maze test. Our data showed that CDK5 knock down induced an increase in p35 protein levels and Rac activity in triple transgenic Alzheimer's mice, which correlated with the recovery of cognitive function; these findings confirm that increased p35 and active Rac are involved in neuroprotection. In summary, our data suggest that p35 acts as a mediator of Rho GTPase activity and contributes to the neuroprotection induced by CDK5 RNAi.

ß-Secretase 1's Targeting Reduces Hyperphosphorilated Tau, Implying Autophagy Actors in 3xTg-AD Mice.

ß-site APP cleaving enzyme 1 (BACE1) initiates APP cleavage, which has been reported to be an inducer of tau pathology by altering proteasome functions in Alzheimer's disease (AD). However, the exact relationship between BACE1 and PHF (Paired Helical Filaments) formation is not clear. In this study, we confirm that BACE1 and Hsc70 are upregulated in the brains of AD patients, and we demonstrate that both proteins show enhanced expression in lipid rafts from AD-affected triple transgenic mouse brains. BACE1 targeting increased Hsc70 levels in the membrane and cytoplasm fractions and downregulated Hsp90 and CHIP in the nucleus in the hippocampi of 3xTg-AD mice. However, these observations occurred in a proteasome-independent manner in vitro. The BACE1miR-induced reduction of soluble hyperphosphorylated tau was associated with a decrease in MAPK activity. However, the BACE1 RNAi-mediated reduction of hyperphosphorylated tau was only blocked by 3-MA (3-methyladenine) in vitro, and it resulted in the increase of Hsc70 and LAMP2 in lipid rafts from hippocampi of 3xTg-AD mice, and upregulation of survival and homeostasis signaling. In summary, our findings suggest that BACE1 silencing neuroprotects reducing soluble hyperphosphorylated tau, modulating certain autophagy-related proteins in aged 3xTg-AD mice.

Silencing of CDK5 as potential therapy for Alzheimer's disease.

Neurodegeneration is one of the greatest public health challenges for the 21st century. Among neurodegenerative diseases, Alzheimer's disease (AD) is the most prevalent and best characterized. Nevertheless, despite the large investment in AD research, currently there is no effective therapeutic option. In the present review, we highlight a novel alternative, which takes advantage of the biotechnological outbreak deployed by the discovery of the RNA interference-based gene silencing mechanism, and its application as a tool for neurodegeneration treatment. Here, we highlight cyclin-dependent kinase 5 (CDK5) as a key candidate target for therapeutic gene silencing. Unlike other members of the cyclin-dependent kinase family, CDK5 does not seem to play a crucial role in cell cycle regulation. By contrast, CDK5 participates in multiple functions during nervous system development and has been established as a key mediator of Tau hyperphosphorylation and neurofibrillary pathology, thus serving as an optimal candidate for targeted therapy in the adult nervous system. We propose that the use of RNA interference for CDK5 silencing presents an attractive and specific therapeutic alternative for AD and perhaps against other tauopathies.

Silencing of CDK5 reduces neurofibrillary tangles in transgenic alzheimer's mice.

Alzheimer's disease is a major cause of dementia for which treatments remain unsatisfactory. Cyclin-dependent kinase 5 (CDK5) is a relevant kinase that has been hypothesized to contribute to the tau pathology. Several classes of chemical inhibitors for CDK5 have been developed, but they generally lack the specificity to distinguish among various ATP-dependent kinases. Therefore, the efficacy of these compounds when tested in animal models cannot definitively be attributed to an effect on CDK5. However, RNA interference (RNAi) targeting of CDK5 is specific and can be used to validate CDK5 as a possible treatment target. We delivered a CDK5 RNAi by lentiviral or adenoassociated viral vectors and analyzed the results in vitro and in vivo. Silencing of CDK5 reduces the phosphorylation of tau in primary neuronal cultures and in the brain of wild-type C57BL/6 mice. Furthermore, the knockdown of CDK5 strongly decreased the number of neurofibrillary tangles in the hippocampi of triple-transgenic mice (3×Tg-AD mice). Our data suggest that this downregulation may be attributable to the reduction of the CDK5 availability in the tissue, without affecting the CDK5 kinase activity. In summary, our findings validate CDK5 as a reasonable therapeutic target for ameliorating tau pathology.