The influence of spin-labeled fluorene compounds on the assembly and toxicity of the aβ peptide

Background

The deposition and oligomerization of amyloid β (Aβ) peptide plays a key role in the pathogenesis of Alzheimer''s disease (AD). Aβ peptide arises from cleavage of the membrane-associated domain of the amyloid precursor protein (APP) by β and γ secretases. Several lines of evidence point to the soluble Aβ oligomer (AβO) as the primary neurotoxic species in the etiology of AD. Recently, we have demonstrated that a class of fluorene molecules specifically disrupts the AβO species.

Methodology/Principal Findings

To achieve a better understanding of the mechanism of action of this disruptive ability, we extend the application of electron paramagnetic resonance (EPR) spectroscopy of site-directed spin labels in the Aβ peptide to investigate the binding and influence of fluorene compounds on AβO structure and dynamics. In addition, we have synthesized a spin-labeled fluorene (SLF) containing a pyrroline nitroxide group that provides both increased cell protection against AβO toxicity and a route to directly observe the binding of the fluorene to the AβO assembly. We also evaluate the ability of fluorenes to target multiple pathological processes involved in the neurodegenerative cascade, such as their ability to block AβO toxicity, scavenge free radicals and diminish the formation of intracellular AβO species.

Conclusions

Fluorene modified with pyrroline nitroxide may be especially useful in counteracting Aβ peptide toxicity, because they posses both antioxidant properties and the ability to disrupt AβO species.     

The Ramberg-Bäcklund reaction for the synthesis of C-glycosides, C-linked-disaccharides and related compounds

The discovery of the Ramberg-B?cklund procedure for preparing exo-glycals from S-glycoside dioxides, developed independently in (Old) York and New York, is reviewed. The methodology is successful with glucose, galactose, mannose, xylose, fucose, ribose, altrose, 2-deoxy-arabino-hexose (2-deoxy-glucose) and daunosamine derivatives, and has been used to prepare di-, tri- and tetra-substituted exo-glycals. More recent developments, such as one-pot variants, and protecting group-free procedures, are also covered. Synthetic applications of the exo-glycals, for example, to prepare beta-glycosidase inhibitors, spirocyclic glucose derivatives, beta-C-glycosides, C-glycosyl porphyrin glycoconjugates and C-glycosyl amino acids, are also discussed. Finally, applications of the Ramberg-B?cklund process for the synthesis of known and novel C-glycosides, and in natural product synthesis, are reviewed.     

Effect of structural modification on the gastrointestinal stability and hepatic metabolism of α-aminoxy peptides

α-Aminoxy peptide AxyP1 has been reported to form synthetic chloride channel in living cells, thus it may have therapeutic potential for the treatment of diseases associated with chloride channel dysfunction. However, this study revealed significant gastrointestinal (GI) instability and extensive hepatic metabolism of AxyP1. To improve its GI and metabolic stability, structural modifications were conducted by replacing the isobutyl side chains of AxyP1 with methyl group (AxyP2), hydroxymethyl group (AxyP3), 4-aminobutyl group (AxyP4) and 3-carboxyl propyl group (AxyP5). Compared with AxyP1 (41 and 47 % degradation), GI stability of the modified peptides was significantly improved by 8-fold (AxyP2), 9-fold (AxyP3) and 12-fold (AxyP5) with no degradation for AxyP4 in simulated gastric fluid within 1 h, and by 12-fold (AxyP2) and 9-fold (AxyP3) with no degradation for AxyP4 and AxyP5 in simulated intestinal fluid within 3 h, respectively. The hepatic metabolic stability of the four modified peptides within 30 min in rat liver S9 preparation was also improved significantly with no metabolism of AxyP5 and threefold (AxyP2 and AxyP4) and eightfold (AxyP3) less metabolism compared with AxyP1 (39 % metabolism). Unlike hydrolysis as the major metabolism of peptides of natural α-amino acids, oxidation mediated by the cytochrome P450 enzymes, especially CYP3A subfamily, to form the corresponding mono-hydroxyl metabolites was the predominant hepatic metabolism of the five α-aminoxy peptides tested. The present findings demonstrate that structural modification can significantly improve the GI and metabolic stability of α-aminoxy peptides and thus increase their potential for therapeutic use in the treatment of chloride channel related diseases.     

Cardiovascular effects of peptides related to the enkephalins and β-casomorphin

In the urethane-anesthetized rat, intravenous injections of morphine produced a short-lasting fall in heart rate and blood pressure. The fall in heart rate (which is vagal in origin) was used here to bioassay peptides related to the enkephalins [-enk] and β-casomorphin [β-C, H-Tyr-Pro-Phe-Pro-Gly-Pro-Ile-OH]. A > 10% decrease in heart rate was used as a quantal index of response and the intravenous ED50 [μmol/kg] were estimated as: methionine-enk, 1.3 ± .24; leucine-enk, 1.5 ± .20; [D-Ala², D-Leu⁵]-enk, 0.45 ± .05; [D-Ala², NMePhe⁴, Met(0)⁵-ol]-enk, 0.0011 ± .0002; H-Tyr-Arg-OH, > 2.0; β-C (1–7), > 2.0; β-C(1–5), > 2.0; β-C(1–4), > 4.0; β-C(1–3), > 2.0; morphine sulfate, 0.11 ± .03; and human β-endorphin,, 0.07 ± .01. One β-C derivative, H-Tyr-Pro-Phe-Pro-NH₂ [β-C(1–4)-NH₂], was active at 0.32 ± .08. Naloxone pretreatment blocked the bradycardia produced by the enkephalins and β-C(1–4)-NH₂. The bioassay described here, based on heart rate, may prove to be useful for the rapid detection and estimation of the $\text{in vivo}$ pharmacological activities of new opioid peptides.     

Theβ-sheets of proteins,the biosynthetic relationships between amino acids,and the origin of the genetic code

Two forces are generally hypothesised as being responsible for conditioning the origin of the organization of the genetic code: the physicochemical properties of amino acids and their biosynthetic relationships (relationships between precursor and product amino acids). If we assume that the biosynthetic relationships between amino acids were fundamental in defining the genetic code, then it is reasonable to expect that the distribution of physicochemical properties among the amino acids in precursor-product relationships cannot be random but must, rather, be affected by some selective constraints imposed by the structure of primitive proteins. Analysis shows that measurements representing the size of amino acids, e.g. bulkiness, are specifically associated to the pairs of amino acids in precursor-product relationships. However, the size of amino acids cannot have been selected per se but, rather, because it reflects the-sheets of proteins which are, therefore, identified as the main adaptive theme promoting the origin of genetic code organization. Whereas there are no traces of the-helix in the genetic code table.The above considerations make it necessary to re-examine the relationship linking the hydrophilicity of the dinucleoside monophosphates of anticodons and the polarity and bulkiness of amino acids. It can be concluded that this relationship seems to be meaningful only between the hydrophilicity of anticodons and the polarity of amino acids. The latter relationship is supposed to have been operative on hairpin structures, ancestors of the tRNA molecule. Moreover, it is on these very structures that the biosynthetic links between precursor and product amino acids might have been achieved, and the interaction between the hydrophilicity of anticodons and the polarity of amino acids might have had a role in the concession of codons (anticodons) from precursors to products.     

The effects of fishing on the diversity,biomass and trophic structure of Seychelles’ reef fish communities

A fishery independent underwater visual census technique was used to assess the effects of fishing on the diversity, biomass and trophic structure of the diurnally active non-cryptic reef-associated fish communities of the Seychelles. One hundred and thirty four species associated with three significantly different types of reef habitat were censused at one unfished ground and in six fishing grounds subject to different fishing intensity and the biomass of several species targeted by the fishery. The diversity of families containing target species (lutjanidae, lethrinidae) was significantly higher at unfished and lightly fished sites as was the total biomass of the fish community and the biomass of piscivorous, piscivorous/invertebrate feeding and herbivorous trophic groups. However, there was no indication that the biomass of non-target species increased in response to the removal of their predators by fishing. The findings of this study are significant for fishery managers because they suggest that the intensive differential cropping of top predators will not necessarily lead to increases in the biomass and productivity of their prey.     

The exogastrula-inducing peptides in embryos of the sea urchin,Anthocidaris crassispina — isolation and determination of the primary structure

Four exogastrula-inducing peptides, A, B, C, and D have been isolated from the homogenates of embryos of the sea urchin, Anthocidaris crassispina, with successive chromatographic fractionations. The complete amino acid sequences of the peptides A and D were determined by analysis of the peptides generated by their digestion with lysyl endopeptidase. They were composed of 52 and 53 amino acid residues, and their molecular weights were calculated to be 5754 and 5737, respectively. The sequences of peptides A and D were DSVYQCNRDTNSCDGFGKCEKSTFGRTTGQYICNCDDGYRNNAYGGCSPRTE, and DTVARCERDTKNCDGHGTCQLSTFGRRTGQYICFCDAGYRKPNSYGGCSPSSA, respectively. The biological significance of the exogastrula-inducing peptides was discussed.     

Effects of curcumin and related compounds on processes involving α-hydroxyethyl radicals

Effects of curcumin and related compounds on product formation in radiolysis of aerated and deaerated ethanol were studied. Ab initio calculations of enthalpy values relating to O-H bond dissociation and H-atom addition to > C = O bonds of the compounds under study have been performed. The obtained data allowed the conclusion that the presence of a 7-carbon chain containing conjugated > C = C < and > C = O bonds in the structures of curcumin and its analogues makes these compounds capable of inhibiting the reactions involving α-hydroxyl-containing carbon-centered radicals. This finding broadens the existing views concerning radical-regulating properties of curcuminoids, and it should be taken into account when practical use of these compounds is envisaged.     

The influence of vegetation pattern on the productivity,diversity and stability of vegetation: The case of `brousse tigrée' in the Sahel

Sample sites of `brousse tigrée' and related vegetation types are described for Mali and Niger. Species composition and physical structure of the herbaceous layer as well as woody plant population were recorded at all sites together with data on soils and natural resource management. Herbage yield was measured whereas foliage yield and wood mass were calculated using allometry equation calibrated for each species. `Brousse tigrée' is characterized by the regularly alternating bare-soil stripes with dense linear thickets arranged perpendicularly to the slope. There was no clear superiority in total plant production of `brousse tigrée' when compared to neighbouring site with diffuse vegetation. However, the pattern of `brousse tigrée' tended to favour woody plant yield to the detriment of herbage yield. The number of herbaceous species recorded per site (22–26) was slightly above Sahelian vegetation average despite low number of species per 1-m² quadrat (6–9), bare soil excluded. This species richness reflects the diversity in edaphic niches resulting from the redistribution and local concentration of water resources and shade. The high spatial heterogeneity and species richness of the herbaceous layer in `brousse tigrée' did not attenuate the interannual variation in herbage yield despite low yields. Except for the herb layer, little evidence was found of grazing influence on the vegetation structure and yield a few hundred metres away from livestock concentration points. On the other hand, the clearing of thickets for cropping led to severe soil erosion which threaten the resilience of `brousse tigrée'. These observations and the well-defined climatic, soiland topographic situations under which the `brousse tigrée' occurs invalidate the hypothesis of an anthropic origin of that vegetationpattern.     

More diversity,less tolerance? The effect of type of cultural diversity on the erosion of tolerance in Swiss municipalities

Immigration and the ways in which host societies receive newcomers pose challenges for modern civil societies. This article contributes to the ongoing discussion about how ethnic diversity influences tolerance towards immigrants. Compared to previous studies, we analyse tolerance as a sequential concept in order to uncover the effects of contextual diversity on attitudes towards immigrants and the granting of certain rights to this group. Moreover, we distinguish different shares of ethnic groups based on their ethnic and cultural origins both on the independent and dependent variable. The analysis relies on a subnational survey of sixty municipalities in Switzerland, revealing that only certain ethnic groups are seen as an economic and cultural threat.     

Functional diversity of bioactive peptides in the nervous system itself: “How the brain may understand”

The interactions involving cells of the nervous system are a complex form of intercellular communication. Biosynthesis of peptide hormones or active neuropeptides is generally through a precursor which provides increased product choices as a function of the processing pathway. Proteolytic processing as well as other molecular modification lead to a wide range of mature products which may vary in different tissues even though they are derived from the same precursor. Also the same neuropeptide may exhibit different bioactivities for different target cells. Finally, by means of collective packaging in secretory organelles, a cell may be able by synergism to further broaden its biologic effects. In these ways, what is seen as added complication in the CNS, may be from the point of view of the cell, a successful attempt to increase its survival ability to adapt and influence its bioenvironment.     

Immunohistochemical demonstration of peptides,serotonin and dopamine-β-hydroxylase-like material in the nervous system of the leechHirudo medicinalis

Summary The central ganglia of the leech,Hirudo medicinalis, were processed for the immunohistochemical localisation of bombesin-, substance P-, cholecystokinin-, vasoactive intestinal polypeptide-, enkephalin-, serotonin- and dopamine--hydroxylase-related substances. To varying extents all of the substances were localised in neuropile processes, and all, with the exception of substance P, were associated with specific perikarya. The most prominent neuropeptides, in terms of the number of immunoreactive neurones, were cholecystokinin and vasoactive intestinal peptide. The dopamine--hydroxylase positive neurones are thought to be octopaminergic, and the serotonin monoclonal antibody revealed positive staining in the Retzius cells. We were unable to demonstrate the coexistence of pairs of substances in any neurones in the leech ganglia.     

A note on sporadic nonperiodic oscillations in the activity of the endocrine system and on the endocrine—Nervous system feedback

By generalizing a previous paper on periodicities in the endocrine system (Bull. Math. Biophysics,30, 735–749, 1968), it is shown that nonperiodic, sporadic oscillations in the system are also possible. A procedure of describing the feedback mechanism between the endocrine system and the central nervous system is suggested. It is shown that the combined system: endocrine—CNS, also may show sporadic fluctuations.     

The stele – a developmental perspective on the diversity and evolution of primary vascular architecture

The stele concept is one of the oldest enduring concepts in plant biology. Here, I review the history of the concept and build an argument for an updated view of steles and their evolution. Studies of stelar organization have generated a widely ranging array of definitions that determine the way we classify steles and construct scenarios about the evolution of stelar architecture. Because at the organismal level biological evolution proceeds by changes in development, concepts of structure need to be grounded in development to be relevant in an evolutionary perspective. For the stele, most traditional definitions that incorporate development have viewed it as the totality of tissues that either originate from procambium – currently the prevailing view – or are bordered by a boundary layer (e.g. endodermis). Consensus between these two perspectives can be reached by recasting the stele as a structural entity of dual nature. Following a brief review of the history of the stele concept, basic terminology related to stelar organization, and traditional classifications of the steles, I revisit boundary layers from the perspective of histogenesis as a dynamic mosaic of developmental domains. I review anatomical and molecular data to explore and reaffirm the importance of boundary layers for stelar organization. Drawing on information from comparative anatomy, developmental regulation, and the fossil record, I propose a stele concept that integrates both the boundary layer and the procambial perspectives, consistent with a dual nature of the stele. This dual stele model posits that stelar architecture is determined at the apical meristem by two major cell fate specification events: a first one that specifies a provascular domain and its boundaries, and a second event that specifies a procambial domain (which will mature into conducting tissues) from cell subpopulations of the provascular domain. If the position and extent of the developmental domains defined by the two events are determined by different concentrations of the same morphogen (most likely auxin), then the distribution of this organizer factor in the shoot apical meristem, as modulated by changes in axis size and the effect of lateral organs, can explain the different stelar configurations documented among tracheophytes. This model provides working hypotheses that incorporate assumptions and generate implications that can be tested empirically. The model also offers criteria for an updated classification of steles in line with current understanding of plant development. In this classification, steles fall into two major categories determined by the configuration of boundary layers: boundary protosteles and boundary siphonosteles, each with subtypes defined by the architecture of the vascular tissues. Validation of the dual stele model and, more generally, in-depth understanding of the regulation of stelar architecture, will necessitate targeted efforts in two areas: (i) the regulation of procambium, vascular tissue, and boundary layer specification in all extant vascular plants, considering that most of the diversity in stelar architecture is hosted by seed-free plants, which are the least explored in terms of developmental regulation; (ii) the configuration of vascular tissues and, especially, boundary layers, in as many extinct lineages as possible.     

Nonenzymatic hydrolysis reactions of adenosine 5′-triphosphate and related compounds: II. Kinetic studies of the hydrolysis of ATP and related compounds with various polyamines

Various structural features of polyamines which are responsible for the acceleration of the hydrolysis of ATP to ADP and P_i at pH 3–4 were surveyed by means of kinetic studies, leading to the following conclusions: 1) The ethyleneimine chain of the polyamines should be as long as possible; 2) the number of methylene carbon atoms between the two adjacent nitrogen atoms of the polyamine has to be two; 3) the terminal groups of the ethyleneimine chain should be primary amino groups.The rate of ATP hydrolysis in the presence of pentaethylenehexamine (pentaen), which possesses the above properties, was found to be 15 times as high as that of hydrolysis at pH 3.5 in the absence of amines. The kinetic data support the previous assumption that there is formation of an ATP-pentaen complex in the hydrolysis reaction. The formation constant of the complex has been calculated to be K = 1.9 × 10⁴M⁻¹ at pH 3.5 and 50°C from the kinetic data. From the temperature dependence of the rates for pentaen or tetraethylenepentamine, the thermodynamic data of these reactions have been obtained.On the other hand, it has been found that pentaen enhances the hydrolysis of GTP and UTP as well as ATP. No phosphate ester bonds of AMP, p-nitrophenylphosphate and α-d-glucose-1-phosphate were hydrolyzed. Therefore, it may be concluded that hydrolysis of the phosphate ester bond with the polyamine is characteristic of ATP, GTP and UTP.