A Diet Containing Rutin Ameliorates Brain Intracellular Redox Homeostasis in a Mouse Model of Alzheimer's Disease.

Quercetin has been studied extensively for its anti-Alzheimer's disease (AD) and anti-aging effects. Our previous studies have found that quercetin and in its glycoside form, rutin, can modulate the proteasome function in neuroblastoma cells. We aimed to explore the effects of quercetin and rutin on intracellular redox homeostasis of the brain (reduced glutathione/oxidized glutathione, GSH/GSSG), its correlation with ß-site APP cleaving enzyme 1 (BACE1) activity, and amyloid precursor protein (APP) expression in transgenic TgAPP mice (bearing human Swedish mutation APP transgene, APPswe). On the basis that BACE1 protein and APP processing are regulated by the ubiquitin-proteasome pathway and that supplementation with GSH protects neurons from proteasome inhibition, we investigated whether a diet containing quercetin or rutin (30 mg/kg/day, 4 weeks) diminishes several early signs of AD. Genotyping analyses of animals were carried out by PCR. In order to determine intracellular redox homeostasis, spectrofluorometric methods were adopted to quantify GSH and GSSG levels using o-phthalaldehyde and the GSH/GSSG ratio was ascertained. Levels of TBARS were determined as a marker of lipid peroxidation. Enzyme activities of SOD, CAT, GR, and GPx were determined in the cortex and hippocampus. ΒACE1 activity was measured by a secretase-specific substrate conjugated to two reporter molecules (EDANS and DABCYL). Gene expression of the main antioxidant enzymes: APP, BACE1, a Disintegrin and metalloproteinase domain-containing protein 10 (ADAM10), caspase-3, caspase-6, and inflammatory cytokines were determined by RT-PCR. First, overexpression of APPswe in TgAPP mice decreased GSH/GSSG ratio, increased malonaldehyde (MDA) levels, and, overall, decreased the main antioxidant enzyme activities in comparison to wild-type (WT) mice. Treatment of TgAPP mice with quercetin or rutin increased GSH/GSSG, diminished MDA levels, and favored the enzyme antioxidant capacity, particularly with rutin. Secondly, both APP expression and BACE1 activity were diminished with quercetin or rutin in TgAPP mice. Regarding ADAM10, it tended to increase in TgAPP mice with rutin treatment. As for caspase-3 expression, TgAPP displayed an increase which was the opposite with rutin. Finally, the increase in expression of the inflammatory markers IL-1ß and IFN-γ in TgAPP mice was lowered by both quercetin and rutin. Collectively, these findings suggest that, of the two flavonoids, rutin may be included in a day-to-day diet as a form of adjuvant therapy in AD.

A Neuroprotective Bovine Colostrum Attenuates Apoptosis in Dexamethasone-Treated MC3T3-E1 Osteoblastic Cells.

Glucocorticoid-induced osteoporosis (GIO) is one of the most common secondary forms of osteoporosis. GIO is partially due to the apoptosis of osteoblasts and osteocytes. In addition, high doses of dexamethasone (DEX), a synthetic glucocorticoid receptor agonist, induces neurodegeneration by initiating inflammatory processes leading to neural apoptosis. Here, a neuroprotective bovine colostrum against glucocorticoid-induced neuronal damage was investigated for its anti-apoptotic activity in glucocorticoid-treated MC3T3-E1 osteoblastic cells. A model of apoptotic osteoblastic cells was developed by exposing MC3T3-E1 cells to DEX (0-700 µM). Colostrum co-treated with DEX was executed at 0.1-5.0 mg/mL. Cell viability was measured for all treatment schedules. Caspase-3 activation was assessed to determine both osteoblast apoptosis under DEX exposure and its potential prevention by colostrum co-treatment. Glutathione reduced (GSH) was measured to determine whether DEX-mediated oxidative stress-driven apoptosis is alleviated by colostrum co-treatment. Western blot was performed to determine the levels of p-ERK1/2, Bcl-XL, Bax, and Hsp70 proteins upon DEX or DEX plus colostrum exposure. Colostrum prevented the decrease in cell viability and the increase in caspase-3 activation and oxidative stress caused by DEX exposure. Cells, upon colostrum co-treated with DEX, exhibited higher levels of p-ERK1/2 and lower levels of Bcl-XL, Bax, and Hsp70. Our data support the notion that colostrum may be able to reduce DEX-induced apoptosis possibly via the activation of the ERK pathway and modulation of the Hsp70 system. We provided preliminary evidence on how bovine colostrum, as a complex and multi-component dairy product, in addition to its neuroprotective action, may affect osteoblastic cell survival undergoing apoptosis.

Curcumin-Piperlongumine Hybrids with a Multitarget Profile Elicit Neuroprotection in In Vitro Models of Oxidative Stress and Hyperphosphorylation.

Curcumin shows a broad spectrum of activities of relevance in the treatment of Alzheimer's disease (AD); however, it is poorly absorbed and is also chemically and metabolically unstable, leading to a very low oral bioavailability. A small library of hybrid compounds designed as curcumin analogues and incorporating the key structural fragment of piperlongumine, a natural neuroinflammation inhibitor, were synthesized by a two-step route that combines a three-component reaction between primary amines, ß-ketoesters and α-haloesters and a base-promoted acylation with cinnamoyl chlorides. These compounds were predicted to have good oral absorption and CNS permeation, had good scavenging properties in the in vitro DPPH experiment and in a cellular assay based on the oxidation of dichlorofluorescin to a fluorescent species. The compounds showed low toxicity in two cellular models, were potent inductors of the Nrf2-ARE phase II antioxidant response, inhibited PHF6 peptide aggregation, closely related to Tau protein aggregation and were active against the LPS-induced inflammatory response. They also afforded neuroprotection against an oxidative insult induced by inhibition of the mitochondrial respiratory chain with the rotenone-oligomycin A combination and against Tau hyperphosphorylation induced by the phosphatase inhibitor okadaic acid. This multitarget pharmacological profile is highly promising in the development of treatments for AD and provides a good hit structure for future optimization efforts.

Potentially inappropriate medication in acute hospitalized elderly patients with polypharmacy: an observational study comparing PRISCUS, STOPP, and Beers criteria.

PURPOSE: To compare the prevalence of potentially inappropriate medication (PIM) in the elderly according to the PRISCUS list, STOPP criteria, and Beers criteria. Secondary, to describe the differences using the three criteria focused on the inappropriate prescription of psychotropic drugs in the elderly. METHODS: A retrospective study was performed at Severo Ochoa University Hospital. The study included 365 patients, aged 80 years and older, living in Madrid, Spain. RESULTS: 93.42% of patients received at least one PIM during hospitalization. Using the PRISCUS list, this changed from 32.6 to 2.7% at discharge. Applying STOPP criteria lowered the percentage from 65.20 to 10.95%, and with Beers criteria from 80.27 to 10.13. Lower Barthel index at admission was associated with an increased relative risk for receiving at least one PIM (OR 1.79, 95% CI 1.15-2.80, p = 0.024) using PRISCUS list as a tool in conjunction with STOPP criteria (OR 1.44, 95% CI 0.89-2.33, p = 0.037). Polypharmacy at admission predicted the presence of PIMs with STOPP criteria (OR 1.74, 95% CI 1.07-2.84, p = 0.001). Regarding psychotropic medicines, 208 patients (56.98%) received at least one psychotropic medicine during hospitalization. A total of 26.30% of patients were treated with psychotropic medicines, detected by the PRISCUS list, and 53.97% and 29.85% with STOPP and Beers, respectively. CONCLUSIONS: Explicit criteria are a useful tool for identifying during hospitalization of the elderly patients. As indicated by the results, new research is needed to carry out an adaptation in our country that includes an evaluation of the strengths of the three tools to decrease PIMs and improve prescription in the elderly.

Peripheral IL-6 Levels but not Sarcopenia Are Predictive of 1-Year Mortality After Hip Fracture in Older Patients.

BACKGROUND: Sarcopenic patients may have an increased risk of poor outcomes after a hip fracture. The objective of this study was to determine whether sarcopenia and a set of biomarkers were potential predictors of 1-year-mortality in older patients after a hip fracture. METHODS: About 150 patients at least 80 years old were hospitalized for the surgical treatment of a hip fracture. The primary outcome measure was the death in the first year after the hip fracture. Sarcopenia was defined at baseline by having both low muscle mass (bioimpedance analysis) and handgrip and using the updated European Working Group on Sarcopenia in Older People (EWGSOP2) definition of probable sarcopenia. Janssen's (J) and Masanés (M) cutoff points were used to define low muscle mass. RESULTS: Mortality 1 year after the hip fracture was 11.5%. In univariate analyses, baseline sarcopenia was not associated with mortality, using neither of the muscle mass cutoff points: 5.9% in sarcopenic (J) versus 12.4% in non-sarcopenic participants (p = .694) and 16% in sarcopenic (M) versus 9.6% in non-sarcopenic participants (p = .285). Probable sarcopenia (EWGSOP2) was not associated with mortality. Peripheral levels of IL-6 at baseline were significantly higher in the group of participants who died in the year after the hip fracture (17.14 ± 16.74 vs 11.42 ± 7.99 pg/mL, p = .026). TNF-α peripheral levels had a nonsignificant trend to be higher in participants who died. No other biomarker was associated with mortality. CONCLUSIONS: Sarcopenia at baseline was not a predictor of 1-year mortality in older patients after a hip fracture. IL-6 was associated with a higher risk of mortality in these patients, regardless of sarcopenia status.

Biomarkers of sarcopenia in very old patients with hip fracture.

BACKGROUND: Hip fracture is both a cause and a consequence of sarcopenia. Older persons with sarcopenia have an increased risk of falling, and the prevalence of sarcopenia may be increased in those who suffer a hip fracture. The aim of this study was to explore potential biomarkers (neuromuscular and peripheral pro-inflammatory and oxidative stress markers) that may be associated with sarcopenia in very old persons with hip fracture. METHODS: We recruited 150 consecutive patients ≥80 years old admitted to an orthogeriatric unit for an osteoporotic hip fracture. Muscle mass was assessed pre-operatively using bioelectrical impedance analysis; Janssen's (J) and Masanés' (M) reference cut-off points were used to define low muscle mass. Muscle strength was assessed with handgrip strength (Jamar's dynamometer). Sarcopenia was defined by having both low muscle mass and strength and using the European Working Group on Sarcopenia in Older People 2 definition of probable sarcopenia (low grip strength). Peripheral markers-pro-inflammatory and oxidative stress parameters-were determined either in the plasma or in the erythrocyte fraction obtained from peripheral whole blood of every patient pre-operatively. RESULTS: Mean age was 87.6 ± 4.9 years, and 78.7% were women. The prevalence of sarcopenia was 11.5% with Janssen's, 34.9% with Masanés' cut-offs, and 93.3% with the European Working Group on Sarcopenia in Older People 2 definition of probable sarcopenia. Among the four pro-inflammatory cytokines tested in plasma, only tumour necrosis factor-α was different (lower) in sarcopenic than in non-sarcopenic participants using both muscle mass cut-offs (J 7.9 ± 6.2 vs. 8.3 ± 5.8, M 6.8 ± 4.7 vs. 9.1 ± 6.2). Erythrocyte glutathione system showed a non-significant tendency to lower glutathione levels and glutathione/oxidized glutathione ratios in sarcopenic participants compared with non-sarcopenic subjects. Catalase activity was also lower in sarcopenic participants (J 2904 ± 1429 vs. 3329 ± 1483, M 3037 ± 1430 vs. 3431 ± 1498). No significant differences were found between groups in chymotrypsin-like activity of the 20S proteasome, superoxide dismutase, glutathione peroxidase and butyrylcholinesterase activity, C-terminal agrin fragment, interferon-γ, or interleukin-1ß. CONCLUSIONS: The prevalence of sarcopenia in patients with hip fracture varies according to the definition and the muscle mass reference cut-off points used. We did not find differences in most neuromuscular, pro-inflammatory, or oxidative stress markers, except for lower peripheral tumour necrosis factor-α levels and catalase activity in sarcopenic participants, which may be markers of an early inflammatory reaction that is hampered in sarcopenic patients.

Several targets involved in Alzheimer's disease amyloidogenesis are affected by morin and isoquercitrin.

Long-term consumption of phytochemicals has been associated with a decreased risk of dementia. The modes of action of two flavonols (morin and isoquercitrin) and two flavanones (hesperidin and neohesperidin) were characterized as single-compound drugs in several Alzheimer's disease (AD)-related assays. First, these phytochemicals were assayed in an amyloid toxicity model (MC65 cells). Second, we examined the activity of the flavonoids in cell-free assays against ß- and γ-secretases and acetylcholinesterase activities, as well as agents able to modify the fibrillogenesis of the amyloid ß-peptide. Additionally, they were assayed against glutamate-induced oxytosis, as scavengers of reactive oxygen species (ROS), as inhibitors of caspase-3, -8 and -9 activation and as modulators of the chymotrypsin-like activity of the ubiquitin-proteasome system. Morin and isoquercitrin, unlike flavanones, exhibited significant activities as ß- and γ-secretase inhibitors, as well as capacity to inhibit Aß aggregation and favor its disaggregation. Flavonols and flavanones showed ROS scavenger activity (P < 0.05), attenuation of caspase-3 and -9 activation (P < 0.05) and restoration of the reduced chymotrypsin-like activity of proteasome 20S (P < 0.05) upon H2O2 exposure of APPswe cells. Flavanones failed to protect against glutamate-induced oxytosis. These findings provide new insight into the anti-amyloidogenic effects of morin and isoquercitrin.

Prevalencia de sarcopenia y características de los sarcopénicos en pacientes mayores de 80 años ingresados por fractura de cadera / Prevalence of sarcopenia and characteristics of sarcopenic subjects in patients over 80 years with hip fracture

Objetivo: calcular la prevalencia de sarcopenia en ancianos ingresados por fractura de cadera (FC) y comparar las características de sarcopénicos y no sarcopénicos. Método: se incluyeron 150 pacientes consecutivos de 80 o más años ingresados por una FC. Se diagnosticó sarcopenia a aquellos con baja masa muscular (bioimpedanciometría, puntos de corte de Janssen y Masanés) y baja fuerza muscular (dinamómetro de Jamar). Se recogieron variables sociodemográficas, cognitivas (Pfeiffer, GDS-Reisberg), funcionales (Barthel, FAC), nutricionales (MNA-SF, índice de masa corporal [IMC], ángulo de fase) y se registró el número de caídas y el número de fármacos. Resultados: edad media: 87,6 ± 4,9 años (78,7% mujeres). La prevalencia de sarcopenia fue del 11,5% (Janssen) y 34,9% (Masanés). Del 77,5% que deambulaba de forma independiente, un 40% había sufrido ≥ 3 caídas. El 38% padecía demencia. Un 80,4% presentaba dependencia leve-moderada y el 14,2% era independiente para actividades básicas de la vida diaria (ABVD). El MNA era compatible con malnutrición en el 12,6% y tomaba ≥ 4 medicamentos el 85,2%. Los pacientes sarcopénicos (Masanés) presentaban índice de masa corporal más bajo (18,6 vs. 24,3, p = 0,003); no se encontraron diferencias entre sarcopénicos y no sarcopénicos en otras variables. No hubo asociación entre el ángulo de fase y la sarcopenia. Conclusiones: hasta un tercio de los pacientes mayores que ingresaron por FC presentan sarcopenia en el momento del ingreso. La prevalencia, en el presente estudio, depende de los puntos de corte usados para definir la baja masa muscular. En contra de lo previsible, los pacientes sarcopénicos con FC muy mayores apenas se diferencian de los no sarcopénicos, salvo por un menor IMC

Aim: to estimate the prevalence of sarcopenia in very old patients admitted to an Orthogeriatric Unit for the treatment of a hip fracture (HF), and to compare characteristics of patients with and without sarcopenia. Methods: one hundred and fifty consecutive patients ≥ 80 years old admitted with HF were included. Sarcopenia was diagnosed with low muscle mass (bioimpedance, using two different cut-off points, Janssen and Masanés) and low grip strength (Jamar's dynamometer). Socio-demographic, nutritional variables (MNA-SF, body mass index [BMI], phase angle), cognitive (Pfeiffer, Reisberg's GDS) and functional variables (Barthel index, FAC) were registered, as well as the number of recent falls and medications on admission. Results: mean age: 87.6 ± 4.9 years (78.7% women). Prevalence of sarcopenia: 11.5% (Janssen's cut-offs) and 34.9% (Masanés cut-offs). Of the 77.5% who had independent ambulation before the fracture, 40% reported three or more recent falls. Before admission, 38% had dementia and 80.4% had mild to moderate dependence to BADL before admission; 14.2% were independent for all BADL. MNA was suggestive of malnutrition in 12.6%, and 85.2% were on four or more prescribed drugs. Sarcopenic (Masanés) had a lower BMI than non-sarcopenic participants (18.6 vs 24.3, p = 0.003), but no other significant differences were found between both groups. Phase angle was also unrelated to sarcopenia status. Conclusions: up to one third of very old patients with HF had sarcopenia on admission. Prevalence varied widely depending on the cut-off points selected to define low muscle mass. Sarcopenic patients in this setting were mostly similar to non-sarcopenic patients, except for a lower BMI

[Prevalence of sarcopenia and characteristics of sarcopenic subjects in patients over 80 years with hip fracture]. / Prevalencia de sarcopenia y características de los sarcopénicos en pacientes mayores de 80 años ingresados por fractura de cadera.

INTRODUCTION: Aim: to estimate the prevalence of sarcopenia in very old patients admitted to an Orthogeriatric Unit for the treatment of a hip fracture (HF), and to compare characteristics of patients with and without sarcopenia. Methods: one hundred and fifty consecutive patients ≥ 80 years old admitted with HF were included. Sarcopenia was diagnosed with low muscle mass (bioimpedance, using two different cut-off points, Janssen and Masanés) and low grip strength (Jamar's dynamometer). Socio-demographic, nutritional variables (MNA-SF, body mass index [BMI], phase angle), cognitive (Pfeiffer, Reisberg's GDS) and functional variables (Barthel index, FAC) were registered, as well as the number of recent falls and medications on admission. Results: mean age: 87.6 ± 4.9 years (78.7% women). Prevalence of sarcopenia: 11.5% (Janssen's cut-offs) and 34.9% (Masanés cut-offs). Of the 77.5% who had independent ambulation before the fracture, 40% reported three or more recent falls. Before admission, 38% had dementia and 80.4% had mild to moderate dependence to BADL before admission; 14.2% were independent for all BADL. MNA was suggestive of malnutrition in 12.6%, and 85.2% were on four or more prescribed drugs. Sarcopenic (Masanés) had a lower BMI than non-sarcopenic participants (18.6 vs 24.3, p = 0.003), but no other significant differences were found between both groups. Phase angle was also unrelated to sarcopenia status. Conclusions: up to one third of very old patients with HF had sarcopenia on admission. Prevalence varied widely depending on the cut-off points selected to define low muscle mass. Sarcopenic patients in this setting were mostly similar to non-sarcopenic patients, except for a lower BMI.

INTRODUCCIÓN: Objetivo: calcular la prevalencia de sarcopenia en ancianos ingresados por fractura de cadera (FC) y comparar las características de sarcopénicos y no sarcopénicos. Método: se incluyeron 150 pacientes consecutivos de 80 o más años ingresados por una FC. Se diagnosticó sarcopenia a aquellos con baja masa muscular (bioimpedanciometría, puntos de corte de Janssen y Masanés) y baja fuerza muscular (dinamómetro de Jamar). Se recogieron variables sociodemográficas, cognitivas (Pfeiffer, GDS-Reisberg), funcionales (Barthel, FAC), nutricionales (MNA-SF, índice de masa corporal [IMC], ángulo de fase) y se registró el número de caídas y el número de fármacos. Resultados: edad media: 87,6 ± 4,9 años (78,7% mujeres). La prevalencia de sarcopenia fue del 11,5% (Janssen) y 34,9% (Masanés). Del 77,5% que deambulaba de forma independiente, un 40% había sufrido ≥ 3 caídas. El 38% padecía demencia. Un 80,4% presentaba dependencia leve-moderada y el 14,2% era independiente para actividades básicas de la vida diaria (ABVD). El MNA era compatible con malnutrición en el 12,6% y tomaba ≥ 4 medicamentos el 85,2%. Los pacientes sarcopénicos (Masanés) presentaban índice de masa corporal más bajo (18,6 vs. 24,3, p = 0,003); no se encontraron diferencias entre sarcopénicos y no sarcopénicos en otras variables. No hubo asociación entre el ángulo de fase y la sarcopenia. Conclusiones: hasta un tercio de los pacientes mayores que ingresaron por FC presentan sarcopenia en el momento del ingreso. La prevalencia, en el presente estudio, depende de los puntos de corte usados para definir la baja masa muscular. En contra de lo previsible, los pacientes sarcopénicos con FC muy mayores apenas se diferencian de los no sarcopénicos, salvo por un menor IMC.

Lichens of Parmelioid Clade as Promising Multitarget Neuroprotective Agents.

Neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease are multifactorial disorders which are increasing in incidence and prevalence over the world without existing effective therapies. The search for new multitarget compounds is the latter therapeutic strategy to address these pathological conditions. Lichens have an important and unknown therapeutic value attributed to their unique secondary metabolites. The aim of this study is to evaluate for the first time the in vitro neuroprotective activities and molecular mechanisms underlying methanol extracts of lichens of the parmelioid clade and to characterize major bioactive secondary metabolites responsible for their pharmacological actions. Of the 15 parmelioid lichen species, our results showed that Parmotrema perlatum and Hypotrachyna formosana methanol extracts exhibited high antioxidant activity as evidenced in ORAC, DPPH, and FRAP assays. Then, SH-SY5Y cells were pretreated with methanol extracts (24 h) followed by Fenton reagent exposure (2 h). Pretreatments with these two more antioxidant methanol lichen extracts increased cell viability, reduced intracellular ROS, prevented oxidative stress biomarkers accumulation, and upregulated antioxidant enzyme (CAT, SOD, GR, and GPx) activity compared to Fenton reagent cells. The neuroprotective activity was much higher for H. formosana than for P. perlatum, even equal to or higher than Trolox (reference compound). Moreover, H. formosana extracts inhibited both AChE and BuChE activities in a concentration dependent manner, and P. perlatum only showed concentration dependent activity against AChE. Finally, chemical composition analysis using TLC and HPLC methods revealed that physodic acid, lividic acid, and lichexanthone are major secondary metabolites in H. formosana and stictic acid and constictic acid are in P. perlatum. These results demonstrated that P. perlatum and, specially, H. formosana are promising multitargeted neuroprotective agents due to their antioxidant and AChE and BuChE inhibition activities.

Neurohormetic responses of quercetin and rutin in a cell line over-expressing the amyloid precursor protein (APPswe cells).

BACKGROUND: Plant secondary metabolites may induce adaptive cellular stress-responses in a variety of cells including neurons at the sub-toxic doses ingested by humans. Such 'neurohormesis' phenomenon, activated by flavonoids such as quercetin or rutin, may involve cell responses driven by modulation of signaling pathways which are responsible for its neuroprotective effects. PURPOSE: We attempt to explore the molecular mechanisms involved in the neurohormetic responses to quercetin and rutin exposure, in a SH-SY5Y cell line which stably overexpresses the amyloid precursor protein (APP) Swedish mutation, based on a biphasic concentration-response relationship for cell viability. METHODS: We examined the impact of both natural compounds, at concentrations in its hormetic range on the following cell parameters: chymotrypsin-like activity of the proteasome system; PARP-1 protein levels and expression and caspase activation; APP processing; and the main endogenous antioxidant enzymes. RESULTS: Proteasome activities following quercetin or rutin treatment were significantly augmented in comparison with non-treated cells. Activity of caspase-3 was significantly attenuated by treatment with quercetin or rutin. Modest increased levels of PARP-1 protein and mRNA transcripts were observed in relation to the mild increase of proteasome activity. Significant reductions of the full-length APP and sAPP protein and APP mRNA levels were related to significant enhancements of α-secretase ADAM-10 protein and mRNA transcripts and significant increases of BACE processing in cells exposed to rutin. Furthermore, quercetin or rutin treatment displayed an overall increase of the four antioxidant enzymes. CONCLUSIONS: The upregulation of the proteasome activity observed upon quercetin or rutin treatment could be afforded by a mild increased of PARP-1. Consequently, targeting the proteasome by quercetin or rutin to enhance its activity in a mild manner could avoid caspase activation. Moreover, it is likely that APP processing of cells upon rutin treatment is mostly driven by the non-amyloidogenic pathway leading to a putative reduction of ßA production. Overall induction of endogenous antioxidant enzymes under quercetin or rutin treatments of APPswe cells might modulate its proteasome activity. We might conclude that quercetin and rutin might exert a neurohormetic cell response affecting various signaling pathways and molecular networks associated with modulation of proteasome function.

In vivo [¹8F] FDG PET imaging reveals that p-chloroamphetamine neurotoxicity is associated with long-term cortical and hippocampal hypometabolism.

PURPOSE: p-Chloroamphetamine (PCA) is a neurotoxin that selectively degenerates the serotonin (5-HT) axon terminals. In order to study the brain metabolic consequences induced by serotonergic denervation, a single dose of PCA (2.5 or 10 mg/kg i.p.) was administered to male adult rats. PROCEDURES: In vivo regional brain metabolism was evaluated 3 and 21 days after PCA (2.5 or 10 mg/kg; i.p.) injection by 2-deoxy-2-[(18)F] fluoro-D-glucose ([(18)F] FDG) positron emission tomography (PET). At day 22, the following markers of neurotoxicity were determined: (a) 5-HT axon terminal lesion by 5-HT transporter (SERT) autoradiography, (b) reactive gliosis by glial fibrillary acidic protein immunohistochemistry, and (c) eventual neurodegeneration by DAPI/Fluoro-Jade C labeling. RESULTS: An average of 20 % reduction of [(18)F] FDG uptake in most brain areas was observed at day 21 under 10 mg/kg PCA treatment. Instead, 2.5 mg/kg PCA only reduced metabolic activity in neocortex. Likewise, the high dose of PCA exerted a strong decrease (>30 %) in SERT density in several 5-HT innervated regions, but no effect was found in midbrain raphe nuclei, the main source of serotonergic neurons. Although PCA induced astroglial activation both in hippocampus and cortex in response to axotomy, no signs of neuronal death in these areas were detected. CONCLUSIONS: Overall, [(18)F] FDG PET revealed that the reduction of the brain metabolic activity induced by PCA is related to 5-HT axon terminal lesion, with no apparent affectation of neuronal viability.

Evaluación de la actividad antioxidante de suplementos dietéticos utilizados para mejorar la fertilidad masculina / Assessment of the antioxidant activity of commercial dietary supplements used to improve male fertility

Objetivo: Evaluar la actividad antioxidante de cinco suplementos dietéticos comerciales (AndroMÁS®, Androferti®, Aquilea Fértil®, GestaDha®, Seidiferty®) utilizados para mejorar la fertilidad masculina. Material y Métodos: La actividad antioxidante se determinó mediante dos técnicas estandarizadas en la literatura para suplementos dietéticos: ensayo ORAC (Oxygen Radical Absorbance Capacity) y el ensayo de captación de radical DPPH (diphenylpicryl-hydrazyl). Se analizaron dos lotes de cada suplemento (n=5/lote). Resultados: Los suplementos AndroMÁS y Seidiferty mostraron actividad antioxidante significativa (p<0,05) en el ensayo ORAC con índices ORAC de 0,25 y 0,24 μmol Trolox/mg suplemento, respectivamente, valores comprendidos en el rango descrito en la literatura para suplementos dietéticos antioxidantes (0,0018-3,18 μmol Trolox/mg suplemento). AndroMÁS, Androferti y Seidifertymostraron una significativa actividad captadora de radical DPPH (p<0,05) con valores de inhibición de 97,2 %, 93,0 % y 60,5 %, respectivamente a una concentración de 0,5 mg/ml. Conclusiones: Este estudio sugiere que las propiedades antioxidantes mostradas por AndroMÁS y Seidiferty pueden contribuir a la eficacia terapéutica demostrada en infertilidad masculina (AU)

Objective: To evaluate the antioxidant activity of five commercially available dietary supplements (AndroMÁS ®, Androferti®, Aquilea Fértil®, GestaDha®, Seidiferty®) used to improve male fertility. Material and Methods: Antioxidant activities were determined using two analytical methods standardized in the literature for dietary supplements namely Oxygen Radical Absorbance Capacity (ORAC) and diphenylpicryl-hydrazyl (DPPH) scavenging radical assays. Two batches of each supplement were analyzed. Results: AndroMÁS and Seidiferty showed significant antioxidant properties (p<0.05) as measured by the ORAC assay, with ORAC values of 0.25 and 0.24 μmol Trolox/mg supplement, within the range reported in the literature for dietary antioxidant supplements (0.018- 3.18 μmol of Trolox equivalent/mg of supplement). The DPPH radical scavenging activity was highest in AndroMÁS, Androferti and Seidiferty (97.2%, 93.0% and 60.5%, respectively) at a concentration of 0.5 mg/ml. Conclusions: This study suggests that the antioxidant properties showed by AndroMÁS and Seidiferty could contribute to the therapeutic efficacy on male infertility (AU)

Processing of the platelet amyloid precursor protein in the mild cognitive impairment (MCI).

It has been suggested that mild cognitive impairment (MCI) patients deteriorate faster than the healthy elderly population and have an increased risk of developing dementia. Certain blood molecular biomarkers have been identified as prognostic markers in Alzheimer's disease (AD). The present study was aimed to assess the status of the platelet amyloid precursor protein (APP) metabolism in MCI and AD subjects and establish to what extent any variation could have a prognostic value suggestive of predictive AD in MCI patients. Thirty-four subjects diagnosed with MCI and 45 subjects with AD were compared to 28 healthy elderly individuals for assessing for protein levels of APP, ß-APP cleaving enzyme 1 (BACE1), presenilin 1 (PS1) and a disintegrin and metalloproteinase-10 (ADAM-10) by western blot, and for the enzyme activities of BACE1 and γ-secretase by using specific fluorogenic substrates, in samples of platelets. A similar pattern in the healthy elderly and MCI patients was found for BACE1 and PS1 levels. A reduction of APP levels in MCI and AD patients compared with healthy elderly individuals was found. Augmented levels of ADAM-10 in both MCI and AD were displayed in comparison with age-matched control subjects. The ratio ADAM-10/BACE1 was higher for the MCI group versus AD group. Whereas BACE1 and PS1 levels were only increased in AD regarding to controls, BACE1 and γ-secretase activities augmented significantly in both MCI and AD groups. Finally, differences and similarities between MCI and AD patients were observed in several markers of platelet APP processing. Larger sample sets from diverse populations need to be analyzed to define a signature for the presence of MCI or AD pathology and to early detect AD at the MCI stage.

Discovery of alkenylboronic acids as neuroprotective agents affecting multiple biological targets involved in Alzheimer's disease.

Alkenylboronic acids have shown important biological activities that contribute to neuroprotection. We have determined their influence on the ß-amyloid (ßA) aggregation process, ß-secretase and acethylcholinesterase activities on cell-free systems, on the redox and lipid peroxidation status, and on the vulnerability to apoptotic death in an APPswe neuroblastoma cell line, before and after hydrogen peroxide treatment. We have discovered that 2-arylvinylboronic acids and some of their esters possess a set of properties which makes them highly useful as neuroprotective agents affecting multiple biological targets involved in AD. These properties are not paralleled by the related 2-arylboronic acids.

SNP-mediated neuroprotection under glucose deprivation is enhanced by Hypericum perforatum.

Hypericum perforatum is a medicinal herb possessing ability for protecting neurons from oxidative stress. Since nitric oxide (NO) may be protective against oxidative stress-induced cell death as occurs in glucose deprivation (GD)-induced neurotoxicity, whether a standardized extract of H. perforatum (HP) increases the NO-mediated neuroprotective effect in GD-PC12 cells was investigated. Induced death in PC12 cells by GD exposure for 18 h was partially prevented by cell incubation with sodium nitroprusside (SNP), a NO-donor. SNP increased survival and nitrite production in GD-cells in a concentration-dependent manner. Co-incubation of cells with 10 µM SNP plus 50-100 µg/ml HP under GD insult significantly prevented GD-induced cell death to a higher extent than SNP alone as shown by an augmentation of cell survival and intracellular bcl-2 levels and a decrease of lipid peroxidation, caspase-3 activation and PARP cleavage. Cytoprotection by the NO-donor was almost abolished by the use of a NO scavenger and potentiated by the presence of superoxide dismutase. SNP and/or HP neuroprotection on GD-cells was significantly reversed by rotenone treatment. These results suggest that: (1) SNP could protect PC12 cells from GD-induced cytotoxicity through NO generation and (2) the enhancement of the SNP-mediated neuroprotective effect on GD-cells by HP might arise in part through scavenging of reactive oxygen species (ROS) and inhibition of mitochondrial dysfunction associated with the hypoglycemic episode. This current finding might highlight the development of therapeutic strategies aimed at manipulating NO-donors in combination with herb supplements containing ROS scavenger compounds for prophylaxis from brain ischemia.

Quercetin and rutin exhibit antiamyloidogenic and fibril-disaggregating effects in vitro and potent antioxidant activity in APPswe cells.

AIMS: Quercetin and rutin have been reported to exert numerous pharmacological activities, such as free-radical scavenging, effects on immune and inflammatory cell functions, and could have benefits in Alzheimer's disease (AD) by mitigating cellular damage induced by reactive oxygen species (ROS). A key event in AD is the conversion of the ß-amyloid (Aß) peptide into amyloid plaques in the brain. Preventing Aß aggregation is pursued as a therapeutic strategy for treating AD. In this study, antiamyloidogenic and antioxidant properties of quercetin and rutin were investigated. MAIN METHODS: We investigated whether quercetin and rutin affect Aß25-35 fibrillogenesis, BACE activity and the cellular redox status. KEY FINDINGS: Quercetin and rutin inhibited the formation of Aß fibrils and disaggregated Aß fibrils. ß-secretase enzyme (BACE) activity was significantly inhibited by rutin. To resemble the in vivo Aß-induced neurotoxicity we used a cell system overexpressing APP Swedish mutation (APPswe), which is associated with early-onset familial AD, and may promote oxidative stress due to the enhanced Aß production. Quercetin and rutin decreased almost completely ROS generation in H(2)O(2)-treated APPswe cells. Both flavonoids increased intracellular GSH content and the redox status, and for rutin this effect was concentration dependent. Besides, quercetin and rutin diminished the index of lipid peroxidation in comparison with control APPswe cells at all concentrations tested. SIGNIFICANCE: Our findings may provide an explanation of the neuroprotective effect of quercetin and rutin, suggesting that they could be dietary phytochemicals able to revert the ß-amyloid toxicity in vivo.

Synthesis and evaluation of arylquinones as BACE1 inhibitors, ß-amyloid peptide aggregation inhibitors, and destabilizers of preformed ß-amyloid fibrils.

BACE1 activity, inhibition of Aß aggregation, and disaggregation of preformed Aß fibrils constitute the three major targets in the development of small-molecule lipophilic new drugs for the treatment of Alzheimer's disease (AD). Quinones are widely distributed among natural products and possess relevant and varied biological activities including antitumor and antibiotic, inhibition of HIV-1 reverse transcriptase, antidiabetic, or COX-inhibition, among others. We report herein the interaction of several arylquinones and their derivatives with the amyloidogenic pathway of the amyloid precursor protein processing. Our studies put forward that these compounds are promising candidates in the development of new drugs which are effective simultaneously towards the three major targets of AD.

In vitro antiamyloidogenic properties of 1,4-naphthoquinones.

The aim of this study is to find out whether several 1,4-naphthoquinones (1,4-NQ) can interact with the amyloidogenic pathway of the amyloid precursor protein processing, particularly targeting at ß-secretase (BACE), as well as at ß-amyloid peptide (Aß) aggregation and disaggregating preformed Aß fibrils. Compounds bearing hydroxyl groups at the quinoid (2) or benzenoid rings (5, 6) as well as some 2- and 3-aryl derivatives (11-15) showed BACE inhibitory activity, without effect on amyloid aggregation or disaggregation. The halogenated compounds 8 and 10 were selective for the inhibition of amyloid aggregation. On the other hand, 1,4-naphthoquinone (1), 6-hydroxy-1,4-naphthoquinone (4) and 2-(3,4-dichlorophenyl)-1,4-naphthoquinone (26) did not show any BACE inhibitory activity but were active on amyloid aggregation and disaggregation preformed Aß fibrils. Juglone (5-hydroxy-1,4-naphthoquinone (3), and 3-(p-hydroxyphenyl)-5-methoxy-1,4-napththoquinone (19) were active on all the three targets. Therefore, we suggest that 1,4-NQ derivatives, specially 3 and 19, should be explored as possible drug candidates or lead compounds for the development of drugs to prevent amyloid aggregation and neurotoxicity in Alzheimer's disease.