Neuroprotective and anti-apoptotic effects of liraglutide on SH-SY5Y cells exposed to methylglyoxal stress.

Glucagon-like peptide 1 (GLP-1) is a growth factor that has demonstrated neuroprotective properties in a range of studies. In an APPswe/PS1ΔE9 mouse model of Alzheimer's disease (AD), we previously found protective effects on memory formation, synaptic plasticity, synapse survival and a reduction of amyloid synthesis and plaque load in the brain. Here, we analyse the neuroprotective properties of the GLP-1 analogue liraglutide in human neuroblastoma cell line SH-SY5Y during methyl glyoxal stress. We show for the first time that cell viability was enhanced by liraglutide (XTT assay) in a dose-dependent way, while cytotoxicity (LDH assay) and apoptosis were reduced. Expression of the pro-survival Mcl1 signaling protein was increased, as was activation of cell survival kinases Akt, MEK1/2 and the transcription factor p90RSK. Liraglutide also decreased pro-apoptotic Bax and Bik expression. In addition, the membrane potential and the influx of calcium into the cell were enhanced by liraglutide. GLP-1 receptor expression was also increased by the drug. The results demonstrate a range of growth factor-related cytoprotective processes induced by liraglutide, which is currently on the market as a treatment for type 2 diabetes (Victoza®). It is also tested in clinical trials in patients with Alzheimer disease.

Development of a high-throughput screen targeting caspase-8-mediated cleavage of the amyloid precursor protein.

Caspases, effectors of apoptosis, are key mediators of neuronal death in several neurodegenerative diseases. Caspase-8 and caspase-6 have been implicated in the pathogenesis of amyotrophic lateral sclerosis, multiple sclerosis, Parkinson's disease, and Alzheimer's disease (AD). ß-Amyloid precursor protein (APP) is cleaved at Asp664 in its intracellular domain by caspase-8. We and other laboratories recently showed that obliteration of the caspase cleavage site on APP alleviates functional AD-like deficits in a mouse model. Therefore, caspase cleavage of APP constitutes a potential novel target for therapeutic intervention. To identify chemical inhibitors of caspase-8 cleavage, we screened a subset of the chemical library at the Harvard NeuroDiscovery Center's Laboratory for Drug Discovery in Neurodegeneration. We show that caspase-8, but not caspase-1, -3, or -9, cleaves a biotinylated peptide derived from APP at Asp664, and we report the development of a sensitive high-throughput assay for caspase-8 cleavage of APP and the use of that assay for the identification of specific small molecule "hit" compounds that potently inhibit Asp664 cleavage of APP. Furthermore, we demonstrate that one of these compounds (LDN-0021835) inhibits the cleavage of APP at Asp664 in cell-based assays.

Cancer cells release glutamate via the cystine/glutamate antiporter.

Although the amino acid glutamate is used as an intercellular signaling molecule for normal bone homeostasis, little is known regarding its possible role in the metabolic disruption characteristic of bone metastasis. We have previously shown in vitro that cancer cell lines relevant to bone metastasis release glutamate into the extracellular environment. This study demonstrates the expression of multiple glutamate transporters in cancer cell lines of non-central nervous system origin. Furthermore, we identify the molecular mechanism responsible for glutamate export and show that this system can be inhibited pharmacologically. By highlighting that glutamate secretion is a common biological feature of cancer cells, this study suggests that tumor-derived glutamate could interfere with glutamate-dependent intercellular signaling in normal bone. Pharmacological interference with cancer cell glutamate release may be a viable option for limiting host bone response to invading tumor cells in bone metastasis.

A by-product of glutathione production in cancer cells may cause disruption in bone metabolic processes.

Bone is a frequent site for metastasis of breast and prostate cancers, often resulting in pathologic changes in bone metabolism and severe pain. The mechanisms involved are not well understood, but tumour cells may release factors that interfere with bone homeostasis. Several observations have led us to hypothesize that the functional disruptions in bone metastasis are the result of a biological process common to many cell types. The high metabolic activity characteristic of cancer cells often upregulates oxidative stress protection mechanisms such as the antioxidant molecule glutathione. In maintaining redox balance, this normal metabolic response may result in unintended pathologic effects in certain sensitive organ sites. Malignant glioma cells kill surrounding neurons in the brain specifically by secreting the amino acid glutamate, an obligatory waste product of glutathione synthesis. We suggest that glutamate release is a plausible mechanism that may account for the pathologic changes in bone metastasis, since bone, like brain, is also highly sensitive to glutamatergic disruption. This report reviews the available evidence to draw a mechanistic connection between tumour cell oxidative stress and the pathology seen in patients with bone metastasis.

Extracellular glutamate alters mature osteoclast and osteoblast functions.

Glutamatergic intercellular communication is involved in many aspects of metabolic homeostasis in normal bone. In bone metastasis, the balance between bone formation and degradation is disrupted. Although the responsible mechanisms are not clear, we have previously identified that cancer cell lines used in bone tumour models secrete glutamate, suggesting that tumour-derived glutamate may disrupt sensitive signalling systems in bone. This study examines the role of glutamate in mature osteoclastic bone resorption, osteoblast differentiation, and bone nodule formation. Glutamate was found to have no effect on the survival or activity of mature osteoclasts, although glutamate transporter inhibition and receptor blockade increased the number of bone resorption pits. Furthermore, transporter inhibition increased the area of resorbed bone while significantly decreasing the number of osteoclasts. Alkaline phosphatase activity and extracellular matrix mineralization were used as measurements of osteoblast differentiation. Glutamate significantly increased osteoblast differentiation and mineralization, but transport inhibitors had no effect. These studies support earlier findings suggesting that glutamate may be more important for osteoclastogenesis than for osteoclast proliferation or functions. Since glutamate is capable of changing the differentiation and activities of both osteoclast and osteoblast cell types in bone, it is reasonable to postulate that tumour-derived glutamate may impact bone homeostasis in bone metastasis.

Vinyl cyanide (CH2CHCN) in interstellar space: potential spectral lines for its detection.

Vinyl cyanide is a molecule having planar geometry with electric dipole moment components, µ a = 3.821 D and µ b = 0.687 D . Thus, a-type transitions are very strong as compared to b-type transitions, and are considered in this investigation. The rotational levels for a-type transitions, may be divided into two distinct groups: one group having odd values of k a and the other having even values of k a . Using the known values of rotational and centrifugal distortion constant along with the electric dipole moment µ a , we have calculated energies of lower 120 rotational levels, having energy up to 92 cm - 1 , for each group, and the probabilities for radiative transitions between the levels. These radiative transition probabilities in conjunction with the scaled values of collisional rate coefficients are used in the Sobolev Large Velocity Gradient analysis. Out of a large number of lines, we have selected the strongest ones. In the low lying levels, besides the 4 observed lines of CH2CHCN, we have discussed about 8 additional transitions, of which 2 showing the phenomenon of anomalous absorption and 6 showing emission feature are found. These lines may play important role for the search of vinyl cyanide in a cosmic object.

Exotic magnetic behaviour and evidence of cluster glass and Griffiths like phase in Heusler alloys Fe2-xMnxCrAl (0 ≤ x ≤ 1).

We present a detailed study of structural, magnetic and thermodynamic properties of a series of Heusler alloys Fe2-xMnxCrAl (x = 0, 0.25, 0.5, 0.75 and 1). Structural investigation of this series is carried out using high resolution synchrotron X-ray diffraction. Results suggest that with increasing Mn concentration, the L21 structure of Fe2CrAl is destabilized. The DC magnetization results show a decrement in paramagnetic (PM) to ferromagnetic (FM) phase transition temperature (TC) with increasing Mn concentration. From the systematic analysis of magnetic memory effect, heat capacity, time dependent magnetization, and DC field dependent AC susceptibility studies it is observed that, Fe2CrAl exhibits cluster glass(CG)-like transition approximately at 3.9 K (Tf2). The alloys, Fe1.75Mn0.25CrAl and Fe1.5Mn0.5CrAl exhibit double CG-like transitions near Tf1 ~ 22 K, Tf2 ~ 4.2 K and Tf1 ~ 30.4 K, Tf2 ~ 9.5 K respectively, however, in Fe1.25Mn0.75CrAl, a single CG-like transition is noted at Tf2 ~ 11.5 K below TC. Interestingly, FeMnCrAl shows the absence of long ranged magnetic ordering and this alloy undergoes three CG-like transitions at ~22 K (Tf*), 16.6 K (Tf1) and 11 K (Tf2). At high temperatures, a detailed analysis of temperature response of inverse DC susceptibility clearly reveals the observation of Griffiths phase (GP) above 300 K (T*) in Fe2CrAl and this phase persists with Mn concentration with a decrement in T*.

Complex magnetic behaviour and evidence of a superspin glass state in the binary intermetallic compound Er5Pd2.

The binary intermetallic compound Er5Pd2 has been investigated using dc and ac magnetic susceptibilities, magnetic memory effect, isothermal magnetization, non-linear dc susceptibility, heat capacity and magnetocaloric effect studies. Interestingly, even though the compound does not show geometrical frustration it undergoes glassy magnetic phase transition below 17.2 K. Investigation of dc magnetization and heat capacity data divulged absence of long-ranged magnetic ordering. Through the magnetic memory effect, time dependent magnetization and ac susceptibility studies it was revealed that the compound undergoes glass-like freezing below 17.2 K. Analysis of frequency dependence of this transition temperature through scaling and Arrhenius law; along with the Mydosh parameter indicate, that the dynamics in Er5Pd2 are due to the presence of strongly interacting superspins rather than individual spins. This phase transition was further investigated by non-linear dc susceptibility and was characterized by static critical exponents γ and δ. Our results indicate that this compound shows the signature of superspin glass at low temperature. Additionally, both conventional and inverse magnetocaloric effect was observed with a large value of magnetic entropy change and relative cooling power. Our results suggest that Er5Pd2 can be classified as a superspin glass system with large magnetocaloric effect.

Enhancement of magnetic ordering temperature and magnetodielectric coupling by hole doping in a multiferroic DyFe0.5Cr0.5O3.

We report the results of our investigation of magnetic, thermodynamic and dielectric properties of Ca substituted half-doped orthochromite, Dy0.6Ca0.4Fe0.5Cr0.5O3. Magnetic susceptibility and heat capacity data bring out that this compound undergoes two antiferromagnetic transitions, one at ~132 and the other at ~22 K. These values are higher than those of DyFe0.5Cr0.5O3. This finding highlights that non-magnetic hole doping in form of Ca+2 in the place of magnetic Dy+3 tends to enhance magnetic transition temperatures in this half-doped orthochromite. We attribute it to possible change in the valence state of Cr/Fe-ion ions due to hole doping. Dielectric anomalies are also seen near the magnetic ordering temperatures indicating magnetodielectric coupling, which is confirmed by magnetic field dependent dielectric studies. The most notable observation is that magnetodielectric coupling strength gets significantly enhanced as compared to DyFe0.5Cr0.5O3. The results reveal that it is possible to tune magnetodielectric coupling by hole doping in this system.

Effect of rare-earth (Er and Gd) substitution on the magnetic and multiferroic properties of DyFe0.5Cr0.5O3.

We report the results of our investigations on the influence of partial substitution of Er and Gd for Dy on the magnetic and magnetoelectric properties of DyFe0.5Cr0.5O3, which is known to be a multiferroic system. Magnetic susceptibility and heat capacity data, apart from confirming the occurrence of magnetic transitions at ~121 and 13 K in DyFe0.5Cr0.5O3, bring out that the lower transition temperature only is suppressed by rare-earth substitution. Multiferroic behavior is found to persist in Dy0.4Ln0.6Fe0.5Cr0.5O3 (Ln = Er and Gd). There is an evidence for magnetoelectric coupling in all these materials with qualitative differences in its behavior as the temperature is changed across these two transitions. Remnant electric polarization is observed for all the compounds. The most notable observation is that electric polarization is seen to get enhanced as a result of rare-earth substitution with respect to that in DyFe0.5Cr0.5O3. Interestingly, a similar trend is seen in the magnetocaloric effect, consistent with the existence of magnetoelectric coupling. The results thus provide evidence for the tuning of magnetoelectric coupling by rare-earth substitution in this family of oxides.

Cancer cell lines release glutamate into the extracellular environment.

Bone is one of the most frequent sites for metastasis of breast and prostate cancers. Bone metastases are associated with pathologic changes in bone turnover and severe pain. The mechanisms that trigger these effects are not well understood, but it is postulated that tumour cells release factors which interfere with signalling processes critical to bone homeostasis. We have identified that several cancer cell lines known to cause bone disruption in animal models of bone metastasis appear to secrete glutamate into their extracellular environment in vitro. Although these cells also express specific glutamate receptors, the implications of this potentially disruptive chemical signal are discussed in relation to normal glutamate-dependent communication processes in bone and a possible mechanistic connection is made between tumour cell glutamate release and the development of pathological changes in bone turnover.

A semi-quantitative GeLC-MS analysis of temporal proteome expression in the emerging nosocomial pathogen Ochrobactrum anthropi.

BACKGROUND: The alpha-Proteobacteria are capable of interaction with eukaryotic cells, with some members, such as Ochrobactrum anthropi, capable of acting as human pathogens. O. anthropi has been the cause of a growing number of hospital-acquired infections; however, little is known about its growth, physiology and metabolism. We used proteomics to investigate how protein expression of this organism changes with time during growth. RESULTS: This first gel-based liquid chromatography-mass spectrometry (GeLC-MS) temporal proteomic analysis of O. anthropi led to the positive identification of 131 proteins. These were functionally classified and physiochemically characterized. Utilizing the emPAI protocol to estimate protein abundance, we assigned molar concentrations to all proteins, and thus were able to identify 19 with significant changes in their expression. Pathway reconstruction led to the identification of a variety of central metabolic pathways, including nucleotide biosynthesis, fatty acid anabolism, glycolysis, TCA cycle and amino acid metabolism. In late phase growth we identified a number of gene products under the control of the oxyR regulon, which is induced in response to oxidative stress and whose protein products have been linked with pathogen survival in response to host immunity reactions. CONCLUSION: This study identified distinct proteomic profiles associated with specific growth points for O. anthropi, while the use of emPAI allowed semi-quantitative analyses of protein expression. It was possible to reconstruct central metabolic pathways and infer unique functional and adaptive processes associated with specific growth phases, thereby resulting in a deeper understanding of the physiology and metabolism of this emerging pathogenic bacterium.