Insoluble Peptides Research Articles

Characterization of monomeric and soluble aggregated Aβ in Down’s syndrome and Alzheimer’s disease brains

AbstractBackgroundAlzheimer’s disease (AD) is the most common form of dementia and is characterized by the two hallmarks, amyloid plaques and neurofibrillary tangles in the brain. The plaques consist of aggregated beta amyloid (Aβ) peptides, formed by proteolytic cleavages of amyloid precursor protein (APP). Down’s syndrome (DS) is a genetic disease with an extra copy of chromosome 21, harbouring the APP gene. DS individuals are at increased risk to develop amyloid pathology. Most DS have substantial amyloid pathology at their 40s, with clinical signs of deterioration before the age of 60. Our aim was to compare the Aβ peptide pattern in DS and AD brains, to better understand the difference between them.MethodsImmunoprecipitation (IP) in combination with mass spectrometry was used to identify both soluble and insoluble Aβ peptides, as well as to characterise the composition of Aβ oligomers/protofibrils. Two different antibody combinations were used; 6E10+4G8 to identify total Aβ peptide contents and the Aβ protofibril selective antibody mAb158 to identify the Aβ peptides present in oligomers/protofibrils. Before IP, cortical tissue from patients with DS (with amyloid pathology), sporadic AD and controls were homogenized and fractionated into TBS (TBS soluble) and formic acid (insoluble) fractions.ResultsFrom IP with 6E10+4G8 several truncated Aβ peptides were identified in all groups, including: 1‐X, 2‐X, 3‐X, 4‐X, 5‐X, 11‐X and –X to 15 as well as post‐translationally modified peptides with oxidation and pyroglutamation. Similarities were observed when comparing the Aβ peptide processing in AD and DS, with the exception of –X to 15 Aβ peptides for which the pattern is different. By using mAb158, Aβ1‐40, Aβ1‐42 and Aβ4‐42 were identified. Higher relative AβX‐42 signals were obtained compared to samples IP’d with 6E10+4G8, indicating that the protofibrils/oligomers were enriched with peptides ending at aa42.ConclusionAll Aβ peptides found in AD are also present in DS (with amyloid pathology) indicating a generally similar pathway of Aβ peptide production, apart from –X to 15. Likewise, the Aβ peptides forming protofibrils/oligomers in both AD and DS were similar, implying the possibility to exploit the potential utility of a treatment with clinical benefit in sporadic AD for subjects with DS.

Rice peptide nanoparticle as a bifunctional food-grade Pickering stabilizer prepared by ultrasonication: Structural characteristics, antioxidant activity, and emulsifying properties

In this study, a novel food-grade Pickering stabilizer was fabricated from insoluble rice peptide aggregates that are considered undesirable and formed during the hydrolysis of rice protein using ultrasonication. The results confirmed that ultrasonication was effective in fabricating rice peptide nanoparticles (RPNs) with a spherical appearance, and the particle size was reduced with ultrasonic time, reaching a minimum size of 357.8 nm in 30 min. Moreover, ultrasonic treatment could improve the antioxidant activity of RPNs by promoting the DPPH scavenging (3.5-fold increase) and Fe2+ chelating activity (3.8-fold increase). Notably, the bioactive RPNs could form stable Pickering emulsions that possess both physical and oxidative stability during storage, which might be due to the antioxidative physical barrier formed by RPNs. These findings suggest a new approach for the effective utilization of insoluble aggregates produced during protein hydrolysis as well as provide a novel bifunctional Pickering stabilizer with intrinsic antioxidant properties.

Human Serum Albumin-Inspired Glycopeptide-Based Multifunctional Inhibitor of Amyloid-β Toxicity.

In Alzheimer’s disease (AD), insoluble Aβ42 peptide fragments self-aggregate and form oligomers and fibrils in the brain, causing neurotoxicity. Further, the presence of redox-active metal ions such as Cu2+ enhances the aggregation process through chelation with these Aβ42 aggregates as well as generation of Aβ42-mediated reactive oxygen species (ROS). Herein, we have adopted a bioinspired strategy to design and develop a multifunctional glycopeptide hybrid molecule (Glupep), which can serve as a potential AD therapeutic. This molecule consists of a natural metal-chelating tetrapeptide motif of human serum albumin (HSA), a β-sheet breaker peptide, and a sugar moiety for better bioavailability. We performed different biophysical and docking experiments, which revealed that Glupep not only associates with Aβ42 but also prevents its self-aggregation to form toxic oligomers and fibrils. Moreover, Glupep was also shown to sequester out Cu2+ from the Aβ–Cu2+ complex, reducing the ROS formation and toxicity. Besides, this study also revealed that Glupep could protect PC12-derived neurons from Aβ–Cu2+-mediated toxicity by reducing intracellular ROS generation and stabilizing the mitochondrial membrane potential. All these exciting features show Glupep to be a potent inhibitor of Aβ42-mediated multifaceted toxicity and a prospective therapeutic lead for AD.

Physicochemical, interfacial and emulsifying properties of insoluble soy peptide aggregate: Effect of homogenization and alkaline-treatment

The physicochemical, interfacial and emulsifying properties of insoluble soy peptide aggregate (ISPA) modified by homogenization and alkaline-treatment (pH10, pH12) were studied in this paper. Homogenization and alkaline-treatment improved the content of hydrophobic amino acids of ISPA, promoting the surface hydrophobicity (H0), decreasing the hydrodynamic diameter (DH) and ζ-potential, finally enhanced the solubility and oil holding capacity (OHC). Compared to ISPA modified at pH10, ISPA modified at pH12 exhibited lower interfacial pressure (π) but higher viscoelastic modulus (E). Emulsions stabilized by modified ISPA had lower ζ-potential, smaller d4,3 and flocculation index (FI), and better stability than untreated ISPA. It was noticed that homogenization pretreatment had a synergistic effect with alkaline treatment on the improvement of emulsifying property. The effects of pH12 on the physicochemical properties of ISPA were more significant than pH10 possibly referred to the disrupting of intramolecular interaction, showing better solubility and emulsifying property. In summary, the emulsifying properties of modified ISPA were even better than that of commercial SPI, indicating that ISPA has the potential to be a good emulsifier in food industry.

Total Chemical Synthesis of ISGylated-Ubiquitin Hybrid Chain Assisted by Acetamidomethyl Derivatives with Dual Functions.

Interferon-stimulated gene 15 (ISG15) is a member of the ubiquitin-like modifiers (ULM) family, which adopts a β-grasp fold domain(s) similar to ubiquitin (Ub) with only minor sequence homology. ISG15 consists of two Ub-like domains and aids the immune system in neutralizing infections by numerous pathogens and plays an important role in defending cells against many viruses including influenza A. Recently, Ub was found to be a substrate for ISG15, which can be ISGylated on Lys29 and Lys48, while the former is more dominant. The discovery of such hybrid ISG15-Ub chains brought forward various fundamental questions regarding the nature and effect of this conjugation. To further investigate the role of hybrid ISG15-Ub chains, the pure homogeneous material of these chains is needed in workable quantities. By applying advanced chemical strategies for protein synthesis, we report the total chemical synthesis of a 231-residue ISG15-Lys29-Ub hybrid chain. During the synthesis we encountered insoluble peptide fragments, and therefore we developed a new reversible Acm based solubilizing tag to efficiently tackle this hurdle. This new Acm tag was compared with the known Arg based Acm solubilizing tag and was found to be more reliable in terms of incorporation and efficiency as demonstrated in the synthesis of the native ISG15-Ub hybrid chain.

Impact of D2O on peptidization of l-hydroxyproline

This is our fourth consecutive study carried out in an order to collect experimental evidence regarding the impact of heavy water (D2O) on the spontaneous oscillatory peptidization of proteinogenic α-amino acids and this time its subject matter is l-hydroxyproline (l-Hyp). Our three earlier studies have been focused on the two sulfur-containing α-amino acids, i.e., l-cysteine (l-Cys) and l-methionine (l-Met), and on one structurally related α-amino acid, i.e., l-proline (l-Pro). It seemed interesting to assess the effect induced by D2O on l-Hyp and to compare it with the effects valid for l-Cys, l-Met, and l-Pro. As analytical techniques, we used high-performance liquid chromatography with the evaporative light-scattering detection (HPLC-ELSD), mass spectrometry (MS), and scanning electron microscopy (SEM). The obtained results make it clear that the impact of heavy water on dynamics of spontaneous peptidization of l-Hyp differs from that exerted on three other α-amino acids discussed (although in all four cases, heavy water significantly hampers spontaneous oscillatory peptidization). With l-Hyp, an increasing proportion of D2O in the reaction mixture results in decreasing yields of both, the soluble lower molecular weight peptides and the insoluble higher molecular weight peptides. With the two sulfur-containing compounds (l-Cys and l-Met), an increasing proportion of D2O in the reaction mixture resulted in growing yields of the soluble lower molecular weight peptides at an expense of decreasing yields of the insoluble higher molecular weight peptides. With l-Pro, still different pattern was observed, namely that the hampering effect of D2O on peptidization was not monotonously dependent on the concentration of D2O in the system, but it was the strongest pronounced for 10% (v/v) D2O in the employed binary methanol–water solvent (with the investigated proportions of D2O in this solvent changing from 0 to 30%). We hope that firm quantitative results presented in this study (and also in the three earlier studies from this cycle) can prove an inspiration for future researchers interested in getting a deeper insight into the role of D2O in life processes, and more specifically in the kinetic and mechanistic aspects thereof. Perhaps it might be noteworthy to add that out of four α-amino acids investigated so far, which can be divided into two groups of the endogenous (l-Cys and l-Pro) and exogenous (l-Met and l-Hyp) species, the endogenous species undergo spontaneous peptidization following the circadian rhythm, whereas the exogenous ones (including l-Hyp) do not.

Cholinesterase-inhibiting Potential and Anti-BACE1 Activities of Edible Leaves of Selected Thai Local Plants

Alzhiemer’s disease (AD) is common amongst the elderly and is associated with decline in brain functions in terms of memory and cognitive loss. The causes of the disease may occur through loss of presynaptic markers of cholinergic system and deposition of amyloid fibrils in the brain. Cholinesterases (ChEs) including acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) are the key enzymes controlling degradation of neurotransmitters, acetylcholines (AChs), in cholinergic hypothesis. Whereas overproduction of b-secretase (BACE1) can generate insoluble b-amyloid peptides. Thus, retardation on enzyme reactions can lead to potential AD prevention. The aim of this research was to investigate in vitro anti-AD activity through key enzymes inhibitions from Thai local plants with edible sour leaves, including Garcinia cowa Roxb., Spondias pinnata (Linn.f.) Kurz, Syzygium gratum (Wight) S.N. Mitra., Tamarind indica L. and Cratoxylum formosum (Jack) Dyer. Leaves were extracted in organic solvents with different polarity index values (ethanol and hexane). As results, all plants possessed different degrees of anti-ChEs activity, in which ethanolic extracts of Spondias pinnata and Tamarind indica exhibited significantly higher ChEs inhibitory activities than Syzygium gratum, Garcinia cowa and Cratoxylum formosum, respectively. Interestingly, most hexane extracts exhibited higher anti-AChE activities than ethanol extracts, while the contrary results were observed in anti-BChE activity. Besides, only Cratoxylum formosum, Garcinia cowa and Tamarind indica extracts possessed anti-BACE1 activity. The information received from this study would be great support of future drug development or nutraceutical agents against AD occurrence regarding its cholinergic and b-amyloid formation hypotheses.

Functionalized Tobacco Mosaic Virus Coat Protein Monomers and Oligomers as Nanocarriers for Anti-Cancer Peptides.

Components with self-assembly properties derived from plant viruses provide the opportunity to design biological nanoscaffolds for the ordered display of agents of diverse nature and with complementing functions. With the aim of designing a functionalized nanoscaffold to target cancer, the coat protein (CP) of Tobacco mosaic virus (TMV) was tested as nanocarrier for an insoluble, highly hydrophobic peptide that targets the transmembrane domain of the Neuropilin-1 (NRP1) receptor in cancer cells. The resulting construct CPL-K (CP-linker-“Kill”) binds to NRP1 in cancer cells and disrupts NRP1 complex formation with PlexA1 as well as downstream Akt survival signaling. The application of CPL-K also inhibits angiogenesis and cell migration. CP was also fused to a peptide that targets the extracellular domain of NRP1 and this fusion protein (CPL-F, CP-Linker-“Find”) is shown to bind to cultured cancer cells and to inhibit NRP1-dependent angiogenesis as well. CPL-K and CPL-F maintain their anti-angiogenic properties upon co-assembly to oligomers/nanoparticles together with CPL. The observations show that the CP of TMV can be employed to generate a functionalized nanoparticle with biological activity. Remarkably, fusion to CPL allowed us to solubilize the highly insoluble transmembrane NRP1 peptide and to retain its anti-angiogenic effect.

Malva parviflora extract ameliorates the deleterious effects of a high\xa0fat diet on the cognitive deficit in a mouse model of Alzheimer\u2019s disease by restoring microglial function via a PPAR-\u03b3-dependent mechanism

BackgroundAlzheimer’s disease (AD) is a neuropathology strongly associated with the activation of inflammatory pathways. Accordingly, inflammation resulting from obesity exacerbates learning and memory deficits in humans and in animal models of AD. Consequently, the long-term use of non-steroidal anti-inflammatory agents diminishes the risk for developing AD, but the side effects produced by these drugs limit their prophylactic use. Thus, plants natural products have become an excellent option for modern therapeutics. Malva parviflora is a plant well known for its anti-inflammatory properties.MethodsThe present study was aimed to determine the anti-inflammatory potential of M. parviflora leaf hydroalcoholic extract (MpHE) on AD pathology in lean and obese transgenic 5XFAD mice, a model of familial AD. The inflammatory response and Amyloid β (Aβ) plaque load in lean and obese 5XFAD mice untreated or treated with MpHE was evaluated by immunolocalization (Iba-1 and GFAP) and RT-qPCR (TNF) assays and thioflavin-S staining, respectively. Spatial learning memory was assessed by the Morris Water Maze behavioral test. Microglia phagocytosis capacity was analyzed in vivo and by ex vivo and in vitro assays, and its activation by morphological changes (phalloidin staining) and expression of CD86, Mgl1, and TREM-2 by RT-qPCR. The mechanism triggered by the MpHE was characterized in microglia primary cultures and ex vivo assays by immunoblot (PPAR-γ) and RT-qPCR (CD36) and in vivo by flow cytometry, using GW9662 (PPAR-γ inhibitor) and pioglitazone (PPAR-γ agonist). The presence of bioactive compounds in the MpHE was determined by HPLC.ResultsMpHE efficiently reduced astrogliosis, the presence of insoluble Aβ peptides in the hippocampus and spatial learning impairments, of both, lean, and obese 5XFAD mice. This was accompanied by microglial cells accumulation around Aβ plaques in the cortex and the hippocampus and decreased expression of M1 inflammatory markers. Consistent with the fact that the MpHE rescued microglia phagocytic capacity via a PPAR-γ/CD36-dependent mechanism, the MpHE possess oleanolic acid and scopoletin as active phytochemicals.ConclusionsM. parviflora suppresses neuroinflammation by inhibiting microglia pro-inflammatory M1 phenotype and promoting microglia phagocytosis. Therefore, M. parviflora phytochemicals represent an alternative to prevent cognitive impairment associated with a metabolic disorder as well as an effective prophylactic candidate for AD progression.

Expression and purification of AtRALF1 from Escherichia coli

Rapid Alakalinization Factors (RALFs) are a family of plant specific peptide growth factors involved in myriad plant processes including growth, development, and response to stresses. These secreted peptides are genetically encoded and processed into an active form from a larger pre-pro-peptide. The active forms are ~5 kDa and have two disulfide bridges. Here we report our attempts to express the active form of AtRALF1 (At1g02900) from the soluble fractions of two different expression strains of E. coli, Origami 2 (DE3) and SHuffleT7 pLysY. Both of these strains are commercially available and genetically altered for expression of proteins with disulfide bonds. Expression of these peptides is challenging because misfolded disulfides or other structural issues result in insoluble peptides. We sought to optimize expression conditions to bulk purify HIS(6x)-AtRALF1 by cobalt column chromatography. Under most conditions the peptide was insoluble. However, at 25oC around 50% of total HIS(6x)-AtRALF1 was soluble in Origami cells. As controls, we expressed mutated versions with alterations in the conserved YISY motif which are physiologically inactive. Surprisingly, these mutant versions were 100% soluble in most conditions tested. All versions of HIS(6x)-AtRALF1 were observed to migrate ~12 kDa in SDS-PAGE gels although their predicted molecular weights is ~8 kDa. Treatment under strong reducing conditions had no effect on this migration pattern. We conclude that AtRALF1’s insolubility issues could be more related to a structure in the YISY motif than to the formation of disulfide bonds.

Unusual confinement properties of a water insoluble small peptide hydrogel.

Unlike polymeric hydrogels, in the case of supramolecular hydrogels, the cross-linked network formation is governed by non-covalent forces. Hence, in these cases, the gelator molecules inside the network retain their characteristic physicochemical properties as no covalent modification is involved. Supramolecular hydrogels thus get dissolved easily in aqueous medium as the dissolution leads to a gain in entropy. Thus, any supramolecular hydrogel, insoluble in bulk water, is beyond the present understanding and hitherto not reported as well. Herein, we present a peptide-based (PyKC) hydrogel which remained insoluble in water for more than a year. Moreover, in the gel state, any movement of solvent or solute to and from the hydrogel is highly restricted resulting in a high degree of compartmentalization. The hydrogel could be re-dissolved in the presence of some biomolecules which makes it a prospective material for in vivo applications. Experimental studies and all atom molecular dynamics simulations revealed that a cysteine containing gelator forms dimers through disulfide linkage which self-assemble into PyKC layers with a distinct PyKC-water interface. The hydrogel is stabilized by intra-molecular hydrogen bonds within the peptide-conjugates and the π-π stacking of the pyrene rings. The unique confinement ability of the hydrogel is attributed to the slow dynamics of water which remains confined in the core region of PyKC via hydrogen bonds. The hydrogen bonds present in the confined water need activation energies to move through the water depleted hydrophobic environment of pyrene rings which significantly reduces water transport across the hydrogel. The compartmentalizing ability is effectively used to protect enzymes for a long time from denaturing agents like urea, heat or methanol. Overall, the presented system shows unique insolubility and confinement properties that could be a milestone in the research of soft-materials.

The metalloprotease ADAMTS4 generates N-truncated Aβ4-x species and marks oligodendrocytes as a source of amyloidogenic peptides in Alzheimer's disease.

Brain accumulation and aggregation of amyloid-β (Aβ) peptides is a critical step in the pathogenesis of Alzheimer's disease (AD). Full-length Aβ peptides (mainly Aβ1-40 and Aβ1-42) are produced through sequential proteolytic cleavage of the amyloid precursor protein (APP) by β- and γ-secretases. However, studies of autopsy brain samples from AD patients have demonstrated that a large fraction of insoluble Aβ peptides are truncated at the N-terminus, with Aβ4-x peptides being particularly abundant. Aβ4-x peptides are highly aggregation prone, but their origin and any proteases involved in their generation are unknown. We have identified a recognition site for the secreted metalloprotease ADAMTS4 (a disintegrin and metalloproteinase with thrombospondin motifs 4) in the Aβ peptide sequence, which facilitates Aβ4-x peptide generation. Inducible overexpression of ADAMTS4 in HEK293 cells resulted in the secretion of Aβ4-40 but unchanged levels of Aβ1-x peptides. In the 5xFAD mouse model of amyloidosis, Aβ4-x peptides were present not only in amyloid plaque cores and vessel walls, but also in white matter structures co-localized with axonal APP. In the ADAMTS4-/- knockout background, Aβ4-40 levels were reduced confirming a pivotal role of ADAMTS4 in vivo. Surprisingly, in the adult murine brain, ADAMTS4 was exclusively expressed in oligodendrocytes. Cultured oligodendrocytes secreted a variety of Aβ species, but Aβ4-40 peptides were absent in cultures derived from ADAMTS4-/- mice indicating that the enzyme was essential for Aβ4-x production in this cell type. These findings establish an enzymatic mechanism for the generation of Aβ4-x peptides. They further identify oligodendrocytes as a source of these highly amyloidogenic Aβ peptides.

Insights into the molecular mechanism behind solubilization of amyloidogenic polyglutamine‐containing peptides

AbstractSynthetic peptides are widely used to understand the protein misfolding and aggregation observed in a diverse group of diseases called systemic amyloidosis and neurodegenerative disorders. The preparation of monomeric solutions of the aggregation‐prone peptides is necessary for achieving a mechanistic understanding efficiently, without compromising any critical step. This study describes novel approaches and mechanisms behind the conversion of otherwise insoluble/sparingly soluble peptides to soluble monomers by trifluoroacetic acid and hexafluoroisopropanol. Although the pretreatment with these solvents results in more or less similar monomers, the mechanism followed to achieve this differs with the solvent. Data show that these solvents aid in the solubility by disrupting the extensive intramolecular and/or intermolecular H‐bond network present in the insoluble peptides.

Cowpea: an overview on its nutritional facts and health benefits.

Cowpea (Vigna unguiculata) is a legume consumed as a high-quality plant protein source in many parts of the world. High protein and carbohydrate contents with a relatively low fat content and a complementary amino acid pattern to that of cereal grains make cowpea an important nutritional food in the human diet. Cowpea has gained more attention recently from consumers and researchers worldwide as a result of its exerted health beneficial properties, including anti-diabetic, anti-cancer, anti-hyperlipidemic, anti-inflammatory and anti-hypertensive properties. Among the mechanisms that have been proposed in the prevention of chronic diseases, the most proven are attributed to the presence of compounds such as soluble and insoluble dietary fiber, phytochemicals, and proteins and peptides in cowpea. However, studies on the anti-cancer and anti-inflammatory properties of cowpea have produced conflicting results. Some studies support a protective effect of cowpea on the progression of cancer and inflammation, whereas others did not reveal any. Because there are only a few studies addressing health-related effects of cowpea consumption, further studies in this area are suggested. In addition, despite the reported favorable effects of cowpea on diabetes, hyperlipidemia and hypertension, a long-term epidemiological study investigating the association between cowpea consumption and diabetes, cardiovascular disease and cancer is also recommended. © 2018 Society of Chemical Industry.

Expression and Purification of Hydrophobic PEP‐Inhibiting Peptides from Bovine αS1 Casein

According to previous studies, there is a prolyl endopeptidase‐inhibiting sequence (LNENLLRFFVAPFPEVFG) within a relatively hydrophobic region of αS1 casein (Juillerat‐Jeanneret et al., 2011). Recently, sixteen recombinant peptides were designed in assorted lengths, each containing the inhibitory sequence at varied positions within the αS1 casein sequence. The corresponding cDNA sequences were originally ligated into the pET15b vector, which provides an N‐terminal poly‐histidine tag for purification. The vector was in turn transformed into a BL21(DE3)pLysS E. coli expression host (Vishram et al., 2016). However, the system proved unsuccessful in adequate expression of the hydrophobic peptide. The intention of this study was to assemble a new pET28a expression system, kindly provided by Dr. X. Cheng (MD Anderson in Houston, TX) that utilizes a SUMO (small ubiquitin‐like modifier) fusion tag. The SUMO tag is known for enhancing expression levels of potentially toxic or insoluble recombinant peptides (Bommarius et al., 2010). The expressed peptides were purified via Ni‐NTA column chromatography, with subsequent UlpI cleavage. This was followed by reversed‐phase high performance liquid chromatography (RP‐HPLC) to further purify the peptides. Tricine‐SDS‐PAGE, a useful screening tool for hydrophobic peptides smaller than 30kDa, was used to analyze expression and purification. A comparison was made between SUMO‐fused expression and non‐fused expression of the hydrophobic casein peptides.Support or Funding InformationOffice of Research and Sponsored Projects, SFASU and Ed & Gwen ColeThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Advances in Synthesis and Application of Nanometer Drug Carriers

The main reason for the formation of nano-biotechnology is due to the penetration of nanotechnology in the biological field, nanotechnology research center is the study of nano-drug carrier. Nano-drug system targeted drug delivery to achieve drug release, increase the insoluble drugs and peptide drug bio-efficiency, reduce the toxicity and application of drugs and other aspects of the development of good prospects, and thus become one of the key research in recent years’ field. Synthesis and application of nanometer drug carriers this review is presented in recent years and its application to provide a comprehensive basis for the treatment process. Describes the nature and preparation of nano-drug carrier methods, in recent years, people have been widely concerned by scholars. Compared with the nano-drug delivery, the general pharmaceutical cannot have to extend the role of drugs, strong efficacy, and the advantages of small drug response. Nano-materials, the specific surface area, surface activity, high catalytic efficiency, surface active center, adsorption capacity and other characteristics, which has many excellent features and new features.

Extra-virgin olive oil ameliorates cognition and neuropathology of the 3xTg mice: role of autophagy.

ObjectiveConsumption of extra virgin olive oil (EVOO), a major component of the Mediterranean diet, has been associated with reduced incidence of Alzheimer's disease (AD). However, the mechanisms involved in this protective action remain to be fully elucidated.MethodsHerein, we investigated the effect of daily consumption of EVOO on the AD‐like phenotype of a mouse mode of the disease with plaques and tangles.ResultsTriple transgenic mice (3xTg) received either regular chow or a chow diet supplemented with EVOO starting at 6 months of age for 6 months, then assessed for the effect of the diet on the AD‐like neuropathology and behavioral changes. Compared with controls, mice receiving the EVOO‐rich diet had an amelioration of their behavioral deficits, and a significant increase in the steady state levels of synaptophysin, a protein marker of synaptic integrity. In addition, they had a significant reduction in insoluble Aβ peptide levels and deposition, lower amount of phosphorylated tau protein at specific epitopes, which were secondary to an activation of cell autophagy.InterpretationTaken together, our findings support a beneficial effect of EVOO consumption on all major features of the AD phenotype (behavioral deficits, synaptic pathology, Aβ and tau neuropathology), and demonstrate that autophagy activation is the mechanism underlying these biological actions.

Turbidity patterns of spontaneous peptidization in an aqueous abiotic system and possible secondary peptide structures

The non-linear dynamics of spontaneous peptidization running in 10 monocomponent and binary abiotic liquid systems of L- and D-Ala and L- and D-Phe is investigated with use of turbidimetry with continuous registration, high-performance liquid chromatography with light scattering detection (HPLC-ELSD), mass spectrometry (MS), and spectroscopy of far UV circular dichroism (CD). The turbidity patterns represent a sum of the light scattering effects caused by insoluble peptides of unknown yields, structures, and molecular weights. The auxiliary analytical techniques confirm the non-linear nature of peptidization (HPLC-ELSD) and spontaneous formation of the homo- and heteropeptides (MS). CD spectroscopy seems to confirm the presence of the secondary α-helix structures. The similarity of turbidity patterns is revealed with the monocomponent (L or D) and binary (L-L or D-D) systems of equichiral α-amino acids, and dissimilarity of patterns is observed with the binary systems of inequichiral α-amino acids (L-D). The tentative conclusion is drawn that the peptides assembled of equichiral α-amino acid units are able to assume the secondary (right- or left-handed α-helix) structures, which in a certain way could foster the similarity of turbidity patterns, and the peptides built of inequichiral α-amino acid units cannot ensure an efficient enough stringing of monomer molecules into equichiral heptades to form complete segments of an α-helix. This randomness of the α-amino acids arrangement in the inequichiral peptide molecules most probably manifests itself as a lack of similarity among the respective turbidity patterns.

Angiotensin-(1–7) administration attenuates Alzheimer’s disease-like neuropathology in rats with streptozotocin-induced diabetes via Mas receptor activation

Diabetes mellitus (DM) is associated with cognitive deficits and an increased risk of Alzheimer’s disease (AD). Recently, a newly identified heptapeptide of the renin-angiotensin system (RAS), angiotensin-(1–7) [Ang-(1–7)], was found to protect against brain damage. This study investigated the effects of Ang-(1–7) on diabetes-induced cognitive deficits. Sprague–Dawley rats were randomly divided into four groups. Diabetes was induced via single i.p. streptozotocin (STZ) injections. Ten weeks after diabetes induction, rats in each group received an intracerebral-ventricular (ICV) infusion of either vehicle, Ang-(1–7) alone, or Ang-(1–7)+A779 daily for two weeks. At the end of the study, Morris water maze (MWM) tests were performed to test cognitive functions before the rats were euthanized. Ang-(1–7) treatment significantly reduced escape latencies in diabetic rats in acquisition trials and markedly enhanced platform area crossing frequency and time spent in the target quadrant in probe trials (3.0±0.39 vs. 1.0±0.33, 39.39±1.11% vs. 25.62±3.07%, respectively, P<0.01). Ang-(1–7) treatment ameliorated damage to the ultrastructure of hippocampal synapses, reduced the expression of hippocampal phospho-tau at Ser396 (P<0.01), Ser404 (P<0.01) and Ser202/Thr205 (P<0.05), and decreased amyloid-β oligomer and both soluble and insoluble β-amyloid peptide 1–42 (Aβ 1–42) and Aβ 1–40 levels (P<0.01). These protective effects were significantly reversed by the co-administration of A779. These findings show that Ang-(1–7) is a promising therapeutic target for diabetes-induced cognitive impairment. The neuroprotective effects of Ang-(1–7) were mainly through Mas receptor (MasR) activation.

The Polyherbal Wattana Formula Displays Anti-Amyloidogenic Properties by Increasing α-Secretase Activities.

Alzheimer’s disease is characterized by the deposition of insoluble amyloid-β peptides produced from the β-amyloid precursor protein (βAPP). Because α-secretase cleavage by ADAM10 and ADAM17 takes place in the middle of Aβ, its activation is considered as a promising anti-AD therapeutic track. Here we establish that the polyherbal Wattana formula (WNF) stimulates sAPPα production in cells of neuronal and non-neuronal origins through an increase of both ADAM10 and ADAM17 catalytic activities with no modification of BACE1 activity and expression. This effect is blocked by specific inhibition or genetic depletion of these disintegrins and we show that WNF up-regulates ADAM10 transcription and ADAM17 maturation. In addition, WNF reduces Aβ40 and Aβ42 generation in human cell lines. Altogether, WNF presents all the characteristics of a potent preventive anti-Alzheimer formula. Importantly, this natural recipe, currently prescribed to patients for the treatment of other symptoms without any secondary effect, can be tested immediately for further clinical studies.