Amylovis-201 enhances physiological memory formation and rescues memory and hippocampal cell loss in a streptozotocin-induced Alzheimer's disease animal model.

Alzheimer's disease is the most common neurodegenerative disease, and its treatment is lacking. In this work, we tested Amylovis-201, a naphthalene-derived compound, as a possible therapeutic candidate for the treatment of AD. For this purpose, we performed three experiments. In the first and third experiment, animals received a bilateral administration of streptozotocin and, starting 24 h after injection, a daily dose of Amylovis-201 (orally), for 17 days or for the whole time of the experiment respectively (28 days), after which learning and memory, as well as the number of hippocampal dentate gyrus cells, were assessed. In the second experiment, healthy animals received a single dose of Amylovis-201, 10 min or 5 h after the learning section to assess whether this substance could promote specific mechanisms involved in memory trace formation. Our data show that, administration of a single dose of Amylovis-201, 10 min after the end of training, but not at 5 h, produces a prolongation in memory duration, probably because it modulates specific mechanisms involved in memory trace consolidation. Furthermore, daily administration of Amylovis-201 to animals with bilateral intracerebroventricular injection of STZ produces a reduction in the loss of the hippocampus dentate gyrus cells and an improvement in spatial memory, probably because Amylovis-201 can interact with some of the protein kinases of the insulin signaling cascade, also involved in neural plasticity, and thereby halt or reverse some of the effects of STZ. Taking to account these results, Amylovis-201 is a good candidate for the therapeutic treatment of AD.

Immune Dysregulation in Autism Spectrum Disorder: What Do We Know about It?

Autism spectrum disorder (ASD) is a group of complex multifactorial neurodevelopmental disorders characterized by a wide and variable set of neuropsychiatric symptoms, including deficits in social communication, narrow and restricted interests, and repetitive behavior. The immune hypothesis is considered to be a major factor contributing to autism pathogenesis, as well as a way to explain the differences of the clinical phenotypes and comorbidities influencing disease course and severity. Evidence highlights a link between immune dysfunction and behavioral traits in autism from several types of evidence found in both cerebrospinal fluid and peripheral blood and their utility to identify autistic subgroups with specific immunophenotypes; underlying behavioral symptoms are also shown. This review summarizes current insights into immune dysfunction in ASD, with particular reference to the impact of immunological factors related to the maternal influence of autism development; comorbidities influencing autism disease course and severity; and others factors with particular relevance, including obesity. Finally, we described main elements of similarities between immunopathology overlapping neurodevelopmental and neurodegenerative disorders, taking as examples autism and Parkinson Disease, respectively.

Motor Coordination Disorders Evaluated through the Grid Test and Changes in the Nigral Nrf2 mRNA Expression in Rats with Pedunculopontine Lesion.

Neurotoxic lesion of the pedunculopontine nucleus (PPN) is known to cause subtle motor dysfunctions. However, motor coordination during advance on a discontinuous and elevated surface has not been studied. It is also not known whether there are changes in the mRNA expression of nuclear factor (erythroid-derived 2)-like 2 (Nrf2) in nigral tissue. METHODS: The effects of the unilateral neurotoxic lesion of the PPN in motor coordination evaluated through grid test and Nrf2 mRNA expression in nigral tissue were evaluated. Two experimental designs (ED) were organized: ED#1 behavioral study (7 and 30 days after PPN lesion) and ED#2 molecular biology study (24 h, 48 h and 7 days) after PPN lesion. RESULTS: ED#1-The number of faults made with left limbs, were significant higher in the lesioned groups (p < 0.01) both 7 and 30 days post-lesion. The number of failures made by the right limbs, was also significantly higher (p < 0.05) vs. control groups. ED#2-Nrf2 mRNA expression showed an increase 24 h after PPN injury (p < 0.01), followed by a peak of expression 48 h post injury (p < 0.001). CONCLUSIONS: Disorders of motor coordination associated with PPN injury are bilateral. The increased Nrf2 mRNA expression could represent an adaptive response to oxidative stress in the nigral tissue following pontine injury.

Nurr1, Pitx3, and α7 nAChRs mRNA Expression in Nigral Tissue of Rats with Pedunculopontine Neurotoxic Lesion.

Background and Objectives: The knowledge that the cholinergic neurons from pedunculopontine nucleus (PPN) are vulnerable to the degeneration in early stages of the Parkinson disease progression has opened new perspectives to the development of experimental model focused in pontine lesions that could increase the risk of nigral degeneration. In this context it is known that PPN lesioned rats exhibit early changes in the gene expression of proteins responsible for dopaminergic homeostasis. At the same time, it is known that nicotinic cholinergic receptors (nAChRs) mediate the excitatory influence of pontine-nigral projection. However, the effect of PPN injury on the expression of transcription factors that modulate dopaminergic neurotransmission in the adult brain as well as the α7 nAChRs gene expression has not been studied. The main objective of the present work was the study of the effects of the unilateral neurotoxic lesion of PPN in nuclear receptor-related factor 1 (Nurr1), paired-like homeodomain transcription factor 3 (Pitx3), and α7 nAChRs mRNA expression in nigral tissue. Materials and Methods: The molecular biology studies were performed by means of RT-PCR. The following experimental groups were organized: Non-treated rats, N-methyl-D-aspartate (NMDA)-lesioned rats, and Sham operated rats. Experimental subjects were sacrificed 24 h, 48 h and seven days after PPN lesion. Results: Nurr1 mRNA expression, showed a significant increase both 24 h (p < 0.001) and 48 h (p < 0.01) after PPN injury. Pitx3 mRNA expression evidenced a significant increase 24 h (p < 0.001) followed by a significant decrease 48 h and seven days after PPN lesion (p < 0.01). Finally, the α7 nAChRs nigral mRNA expression remained significantly diminished 24 h, 48 h (p < 0.001), and 7 days (p < 0.01) after PPN neurotoxic injury. Conclusion: Taking together these modifications could represent early warning signals and could be the preamble to nigral neurodegeneration events.

Tyrosine Hydroxylase, Vesicular Monoamine Transporter and Dopamine Transporter mRNA Expression in Nigrostriatal Tissue of Rats with Pedunculopontine Neurotoxic Lesion.

BACKGROUND: The degeneration of the pedunculopontine nucleus (PPN) precedes the degeneration of the nigral cells in the pre-symptomatic stages of Parkinson's disease (PD). Although the literature recognizes that a lesion of the PPN increases the vulnerability of dopaminergic cells, it is unknown if this risk is associated with the loss of capability of handling the dopaminergic function. METHODS: In this paper, the effects of a unilateral neurotoxic lesion of the PPN in tyrosine hydroxylase (TH), vesicular monoamine transporter 2 (VMAT2) and dopamine transporter (DAT) mRNA expression in nigrostriatal tissue were evaluated. Three experimental groups were organized: non-treated rats, NMDA-lesioned rats and Sham-operated rats. RESULTS: Seven days after the PPN lesion, in nigral tissue, TH mRNA expression was higher in comparison with control groups (p < 0.05); in contrast, VMAT2 mRNA expression showed a significant decrease (p < 0.01). DAT mRNA expression showed a significant decrease (p < 0.001) in the striatal tissue. Comparing nigral neuronal density of injured and control rats revealed no significant difference seven days post-PPN injury. CONCLUSIONS: Findings suggest that the PPN lesion modifies the mRNA expression of the proteins associated with dopaminergic homeostasis at nigrostriatal level. It could represent vulnerability signals for nigral dopaminergic cells and further increase the risk of degeneration of these cells.

Motor dysfunction and alterations in glutathione concentration, cholinesterase activity, and BDNF expression in substantia nigra pars compacta in rats with pedunculopontine lesion.

Pedunculopontine nucleus (PPN) has been considered a critically important region in the regulation of some of the physiological functions that fail during the progression of Parkinson's disease (PD). In this paper, the effects of unilateral neurotoxic lesion of the PPN [through the injection of N-methyl-d-aspartate (NMDA) solution (concentration: 0.1M; volume: 0.5µL)] in motor execution and gait disorders and the changes in cellular and molecular indicators in rat nigral tissue were evaluated. The motor execution was assessed using the beam test (BT) and the gait disorders by footprint test. Glutathione (GSH) concentrations, acetyl cholinesterase enzymatic activity (AChE EA), and brain-derived neurotrophic factor (BDNF) mRNA expression in nigral tissue were analyzed. NMDA-lesioned rats showed fine motor dysfunction with a significant increase in the slow (p≤0.01) and fast movement (p≤0.01) time and in path deviation (p≤0.01) on the smaller diameter beams. Moreover, NMDA-lesioned rats exhibited an imprecise path with moments of advances and setbacks, alternating with left and right deviations, suspensions, and inverted positions. Footprint test revealed slight gait disorders, which were manifested by a reduction in the left and right stride lengths, the intra-step distance, and the support area (p≤0.01). Biochemical studies showed that 48h after the PPN neurotoxic injury, the GSH concentrations and BDNF expression were significantly increased (p≤0.01). These variables returned to normal values 7days after the PPN lesion; the AChE EA showed a significant increase at this time. These functional changes in nigral tissue could be a plastic responses associated with early PD.

The trypsin inhibitor panulirin regulates the prophenoloxidase-activating system in the spiny lobster Panulirus argus.

The melanization reaction promoted by the prophenoloxidase-activating system is an essential defense response in invertebrates subjected to regulatory mechanisms that are still not fully understood. We report here the finding and characterization of a novel trypsin inhibitor, named panulirin, isolated from the hemocytes of the spiny lobster Panulirus argus with regulatory functions on the melanization cascade. Panulirin is a cationic peptide (pI 9.5) composed of 48 amino acid residues (5.3 kDa), with six cysteine residues forming disulfide bridges. Its primary sequence was determined by combining Edman degradation/N-terminal sequencing and electrospray ionization-MS/MS spectrometry. The low amino acid sequence similarity with known proteins indicates that it represents a new family of peptidase inhibitors. Panulirin is a competitive and reversible tight-binding inhibitor of trypsin (Ki = 8.6 nm) with a notable specificity because it does not inhibit serine peptidases such as subtilisin, elastase, chymotrypsin, thrombin, and plasmin. The removal of panulirin from the lobster hemocyte lysate leads to an increase in phenoloxidase response to LPS. Likewise, the addition of increasing concentrations of panulirin to a lobster hemocyte lysate, previously depleted of trypsin-inhibitory activity, decreased the phenoloxidase response to LPS in a concentration-dependent fashion. These results indicate that panulirin is implicated in the regulation of the melanization cascade in P. argus by inhibiting peptidase(s) in the pathway toward the activation of the prophenoloxidase enzyme.

Search of new Antimitotics compounds from the Cuban flora / Búsqueda de nuevos compuestos con actividad antimitótica en la flora cubana

The main objective of anti-carcinogenic chemotherapy is to stop uncontrolled cellular proliferation. This has prompted us to begin a systematic survey of new effective inhibitors with ability to react with cytoskeletal components and arrest living, dividing cells. Even for traditional populations herbs-consuming, encouraging the use of species with chemopreventive actions could be helpful as part of life expectancy improvement strategies. Herbal products have significantly lower costs, exhibit little or no toxicity during long-term oral administration and are relatively available at large scale. Current work involved the screening of 85 extracts from Cuban medicinal plants, selected on the basis of traditional use, ethnobotanics and pharmacological information (antiparasitic, antitumour, abortive, etc.). Antitubulinic activity in the hydroalcoholics extracts was evaluated by using a modified version of the conventional turbidity assay of tubulin assembly/ disassembly. The activity limits of the news isolated antitubulin agents were thoroughly investigated. According to the presented results, the extracts displaying the highest antitubulinic activity were Tamarindus indica L., Lawsonia inermes L and Xanthium strumarium L.

Detener la proliferación celular es el principal propósito de la quimioterapia anticarcinogénica. Para ello se ha realizado una búsqueda a partir de fuentes naturales de nuevos inhibidores efectivos que reaccionen con los componentes del citoesqueleto y puedan detener la división celular. En poblaciones que tradicionalmente utilizan plantas medicinales se estimula el uso de aquellas especies con acción quimiopreventivas como parte de una estrategia que contribuya a la calidad de vida. Los productos herbarios tienen costos significativamente más bajos, exhiben poca o ninguna toxicidad durante la administración oral a largo plazo y están al alcance de todos. Nuestro trabajo consistió en realizar un tamizaje de 85 extractos de plantas medicinales cubanas seleccionadas en base al uso tradicional, en las encuestas etnobotánicas e información farmacológica (actividad antiparasitaria, antitumoral, abortiva, etc). La actividad antitubulínica fue evaluada mediante una versión modificada del ensayo turbimétrico del ensamblaje/desensamblaje de la tubulina. Se determinó la actividad límite de los nuevos agentes antitubulínicos siendo los extractos de Tamarindus indica L., Lawsonia inermes L and Xanthium strumarium L. los de mejor actividad antitubulínica según las condiciones ensayadas.

Genotoxicidad de Justicia pectoralis Jacq: (tilo) / Genotocixity of Justicia pectoralis Jacq: (tilo)

Objetivo: determinar el efecto genotóxico del extracto hidroalcohólico 30 por ciento de partes aéreas de Justicia pectoralis Jacq. (variedad pectoralis) secado por spray dryer. Métodos: se empleó el ensayo Salmonella/microsomas con las líneas TA 1535, TA 1537, TA 98 y TA 100 con un rango de concentraciones de 50 a 5 000 mg de polvo/placa (± S9) según el protocolo de incorporación en placas; para el ensayo de micronúcleos en médula ósea se utilizaron ratones Cenp: NMRI de los 2 sexos, que recibieron dosis isovolumétricas (10 mL/kg) de 500, 1 000 y 2 000 mg de polvo/kg, por vía intragástrica, separadas 24 h, con sacrificio 24 h después de la última aplicación. Resultados: no se encontró efecto genotóxico con ninguna de las cepas en el ensayo de Salmonella/microsomas y el extracto no causó un aumento estadísticamente significativo en la frecuencia de eritrocitos policromáticos micronucleados en los ratones tratados y no mostró relación dosis respuesta positiva. Conclusiones: el polvo no reveló efecto genotóxico en las condiciones experimentales de este estudio.

Objective: to detemine the genotoxic effect of the hydroalcoholic extract 30 of aerial parts of Justicia pectoralis Jacq. (variety pectoralis) dried by spray drier. Methods: the Salmonella/microsomes with lines TA 1535, TA 1537, TA 98 and TA 100 with a concentration range from 50 to 5 000 ìg of powder/plaque (± S9) was used according to the protocol of incorporation in plaques; for the bone marrow micronucleus assay there were used Cenp mice: NMRI of both sexes that received isovolumetric doses (10 mL/kg) of 500, 1 000 y 2 000 mg of powder/kg by intragastric route at intervals of 24 h and were sacrificed 24 h after the last application. Results: no genotoxic effect was found with any of the strains in the assay of Salmonella/microsomes. The extract did not cause a statistically significant increase in the frequency of micronucleated polychromatic erythrocytes in the mice treated. No dose/positive response relation was observed. Conclusions: the powder did not reveal genotoxic effect under the experimental conditions of this.

Genotoxicidad de Indigofera suffruticosa Mill / Genotocixity of Indigofera suffruticosa Mill

Se estudió el efecto genotóxico de un extracto de Indigofera suffruticosa Mill. mediante el ensayo Salmonella/microsomas con las líneas TA 1535, TA 1537, TA 98 y TA 100 que resultó positivo para el protocolo de incorporación en placas con las cepas TA 1535, TA 1537 con un rango de concentraciones de 50 a 5000 µg/placa (±S9). Mediante el ensayo de inducción de micronúcleos en médula ósea de ratón el extracto exhibió un aumento estadísticamente significativo en la frecuencia de PCE micronucleados en las hembras tratadas y una relación dosis respuesta positiva, cuando se evaluaron dosis de 500, 1000 y 2000 mg/kg por vía oral con el ensayo de micronúcleos en médula ósea de ratón.

It was accomplished the study of the genotoxic effect of extract of Indigofera suffruticosa Mill. using the Salmonella/microsoma assay in strains TA 1535, TA 1537, TA 98 and TA 100 was positive in a plate incorporation protocol with concentrations ranging from 50 to 5000 mg/plate (¡À S9) for the strains TA 1535, TA 1537. The extract of Indigofera suffruticosa Mill. exhibited the significantly increment in the frequency of micronucleated polychromatic erythrocytes in female mice and dose-dependent genotoxicity to mouse bone marrow micronucleus test at the doses of 500, 1000 and 2000 mg/kg assayed.

Evaluación genotóxica del extracto hidroalcohólico de Tamarindus indica L. empleando el ensayo de micronúcleos en médula ósea de ratón / Genotoxic assessment of hydroalcoholic extract of Tamarindus indica L using micronuclei assay in mouse bone marrow

El ensayo de micronúcleos en médula ósea de ratón, prueba incluida en la batería estándar de genotoxicidad, fue empleado para evaluar el potencial clastogénico y aneugénico de un extracto hidroalcohólico de Tamarindus indica L. Se utilizaron ratones NMRI de ambos sexos los que fueron administrados por vía intragástrica a razón de 10 mL/kg, separados 24 h, con sacrificio 24 h después de la última aplicación Se determinó el índice de genotoxicidad, porcentaje de eritrocitos policromáticos micronucleados en un total de 1 000 eritrocitos policromáticos por animal. Los índices de genotoxicidad de los grupos tratados con 500, 1000 y 2000 mg/kg de peso corporal del extracto, fueron estadísticamente significativos con respecto al grupo tratado con la sustancia control (H2O estéril). El extracto de Tamarindus indica L. resultó clastogénico, ya que produjo un aumento significativo en la frecuencia de micronúcleos de los eritrocitos policromáticos de la médula ósea y en la prueba de tendencia lineal de proporciones demostró una relación dosis-respuesta.

Micronuclei assay in mouse bone marrow, test included in genotoxicity standard battery, was used to assess clastogenic and aneugenic potential of a hydroalcoholic extract of Tamarindus indica L. We used NMRI mice of both sexes, with administration by intragastric route at a rate of 10 mL/kg, in the space of 24 hours, sacrified 24 hours after last application. Genotoxicity rate, polychromatic micronucleated erythrocyte percentage in a total of 1 000 polychromatic erythrocytes per animal. Genotoxicity rates of treated groups using 500, 1000, and 2000 mg/kg extract/body weight were statistically significant as for group treated with control substance (H2O sterile). Extract of Tamarindus indica L was of clastogenic type, since give raise to a significant increase in micronuclei frequency of bone marrow polychromatic erythrocytes, and in ratios linear trend test showed a dosis-response relationship.