Cns & Neurological Disorders-drug Targets, Oct 18, 2016
Exposure to environmental extremely low-frequency electromagnetic fields (ELF-EMF) in everyday li... more Exposure to environmental extremely low-frequency electromagnetic fields (ELF-EMF) in everyday life is increasing and it is a matter of great debate whether exposure to ELF-EMF can be harmful to human health. The neuropathology and symptoms of neurodegenerative disease depends on factors other than genetic predispositions, such as environmental exposure to disease-related risk factors. Research focusing on a possible contribution of ELF-EMF to cell injury and to the development of neurodegenerative disorders is characterized by conflicting data from epidemiological and animal studies. Due to lack of a direct link between neurodegenerative processes and ELF-EMF exposure, our goal was to investigate if ELF-EMF exposure may represent a possible risk factor. In the present study, using neuronal-like SH-SY5Y neuroblastoma cells, we show that the balance between generation and elimination of reactive oxygen species, as well as the balance between pro- and anti-inflammatory cytokines linked to oxidative stress, was maintained ensuring that cells respond properly to ELF-EMF (50Hz /1mT). In SH-SY5Y-exposed cells we observed increased intracellular 5-hydroxyindoleacetic acid/5-hydroxytryptamine ratio reflecting the rate of transmitter synthesis, catabolism and release, while matrix metalloproteinases that play critical roles in neuronal cell death were not significantly altered. The results presented here indicate that changes caused by short (1h-3h) and sub-chronic (48 h) exposure to 50Hz/1mT ELF-EMF in SH-SY5Y cells are minor in comparison to the neuronal cell damage expected to underlie neurodegeneration or cognitive impairment. Thus, these results are in accord with epidemiological studies that have provided little support for a link between ELF-EMFs and neurodegeneration.
Mesenchymal stem cells (MSCs), are multipotent cells present in several tissues and organs as bon... more Mesenchymal stem cells (MSCs), are multipotent cells present in several tissues and organs as bone marrow and adipose tissue. MSCs derived from Adipose tissue (ASCs) are an attractive source for regenerative medicine because they can be easily isolated, rapidly expandable in culture and show excellent differentiation potential. Acetylcholine (ACh), the most important neurotransmitter in central (CNS) and peripheral nervous system (PNS), plays a key role in the control of physiological processes also in non-neural tissues. Previously demonstrated that M2 muscarinic receptors negatively modulate ASC proliferation and migration. In the present work we demonstrate that rat ASCs also express \u3b17 nAChR receptors. By using the selective \u3b17 nicotinic receptor agonist ICH-3, we demonstrated that \u3b17 receptor activation causes a reduction of cell proliferation without affecting cell survival and morphology. Moreover, by wound healing assay, we have also demonstrated that \u3b17 promotes cell migration and a significant upregulation of CXCR4 receptor. Interestingly the activation of \u3b17 nAChR appears also up-regulate the expression of M2 acetylcholine receptor, suggesting a feedback positive loop between muscarinic and nicotinic receptors. Our data suggest that ACh, via cholinergic receptors might contribute to modulate ASC physiology. Moreover, in particular via nicotinic receptors, ACh might control ASCs migration
Glioblastomas are the most aggressive brain tumors in humans. They show a marked cell heterogenei... more Glioblastomas are the most aggressive brain tumors in humans. They show a marked cell heterogeneity; in particular glioblastoma stem cells (GCSs), a multipotent cell subpopulation capable of self-renewals, have been identified in these tumors. Considering that these cells show high resistance to chemiotherapy, the identification of drugs able to inhibit GCS proliferation and survival is considered of great clinical relevance. The involvement of muscarinic receptors in cancer has been reported. Recently we have demonstrated that the activation of M2 muscarinic receptors, through the agonist arecaidine, arrests cell proliferation and induces apoptosis in primary and stable glioblastoma cell lines. Following previous data, we have investigated the effect produced by M2 stimulation on GCS proliferation. The expression of M2 receptors was analyzed in several GCS cell lines derived from human biopsies. Considering the M2 receptor levels, we have selected two cell lines (GB7 and GB8). In both cell lines the treatment with the M2 agonist arecaidine decreases cell proliferation; in particular in GB7 cells the decrease of cell proliferation is time and dose dependent. Moreover co-treatment of GB7 cells with M1 and M3 antagonists (pirenzepine and 4-DAMP respectively) has indicated that the decreased cell proliferation\u2013arecaidine induced was probably dependent on M2 activation. FACS analysis has also shown that in GB7 cells, arecaidine inhibits cell cycle progression. Conversely in GB8 cells, arecaidine causes a decreased cell survival. The analysis of EGFR and Notch expression has clearly indicated that at least in GB7 cells the decrease of EGFR expression may be responsible on the decreased cell proliferation induced by arecaidine. Considering the previous data, the use of a more selective agonist for M2 receptor may have a strategic relevance for the glioblastoma therapy. As the difficulty of finding ligands with high selectivity, the exploitation of allosteric sites on G protein-coupled receptors (GPCRs) opens up a new range of possibilities in the research of selective orthosteric ligands in particular for GPCRs as mAChRs, which are characterized by highly conserved orthosteric sites among different receptor subtypes. Actually we are testing the effects produced by a new generation of dualsteric ligands for M2 receptors on glioblastoma cell proliferation both in stable cell lines and in the GCSs
Introduction. The involvement of muscarinic acetylcholine receptors (mAChRs) in cancer has been l... more Introduction. The involvement of muscarinic acetylcholine receptors (mAChRs) in cancer has been largely documented. Recently, we have demonstrated that activation of M2AChRs arrests cell proliferation and induces apoptosis in glioblastoma (GB) cell lines and in glioblastoma cancer stem cells (GSC). In the present work, we compared the effects mediated by the preferential activation of M2AChRs, through the M2 agonist arecaidine propargyl ester (APE) and the dualsteric agonist N8-Iper, both in GB cell lines and in GSC, analyzing their antiproliferative and pro-apoptotic effects and their potential use in combined therapy with conventional drugs (i.e., temozolomide). Material and Methods. U251 established cell lines and GSC (GB7 cell line) obtained from human biopsy were used as cell models. MTT assay, trypan blue staining and FACS analysis were used to evaluate cell viability and cell death. The M2 silencing (by siRNA) and pharmacological competition were used to confirm the ability of M2 agonists to selectively bind this receptor subtype. Transcript levels for multidrug efflux pumps (e.g., ATP binding cassette, ABC) were analyzed by Real-Time PCR and Western Blot analysis. Results. In GB7 cells, treatment with the N8-Iper decreased cell proliferation in a time and dose dependent manner as previously demonstrated after APE treatments. FACS analysis has also shown that in GB7 cells N8-Iper caused a significant increase of cell death. The pharmacological competition with M1 and M3 antagonists (pirenzepine and 4-DAMP, respectively) and the M2 antagonist methoctramine indicated that the effects of N8-Iper were dependent on selective activation of the M2AChR subtype. The ability of APE and N-8-Iper to activate M2 receptor populations was also confirmed by CHRM2 siRNA transfection. Moreover, APE and N-8-Iper decreased the mRNA levels and the protein expression of the ABC-G2 pump. Finally, the combined treatment with an M2 agonist and temozolomide indicated a synergic effect of the two drugs in decreasing GB cell survival. Conclusions. Our findings suggest that M2AChR agonists represent a new putative therapeutic tool in glioblastoma treatment. Moreover, the ability of M2 agonists to decrease the ABC-G2 drug efflux pumps expression in GSC cells, coupled with their synergic effect in combination with temozolomide, are indicative of their relevant role in reducing the GB chemoresistance
Neural tissue is a hierarchical multiscale system with intracellular and extracellular diffusion ... more Neural tissue is a hierarchical multiscale system with intracellular and extracellular diffusion compartments at different length scales. The normal diffusion of bulk water in tissues is not able to detect the specific features of a complex system, providing nonlocal, diffusion measurement averaged on a 10-20 μm length scale. Being able to probe tissues with sub-micrometric diffusion length and quantify new local parameters, transient anomalous diffusion (tAD) would dramatically increase the diagnostic potential of diffusion MRI (DMRI) in detecting collective and sub-micro architectural changes of human tissues due to pathological damage. In DMRI, the use of tAD parameters quantified using specific DMRI acquisition protocols and their interpretation has often aroused skepticism. Although the derived formulas may accurately fit experimental diffusion-weighted data, the relationships between the postulated dynamical feature and the underlying geometrical structure remains elusive, or ...
The International journal of developmental biology, 2000
Development of the nervous system is dependent on the co-operation between cell determination eve... more Development of the nervous system is dependent on the co-operation between cell determination events and the action of epigenetic factors; in addition to well known factors, e.g. growth factors, neurotransmitters have been assigned a role as "morphogens" and modulators of neuronal differentiation in an early developmental phase. The possible role of acetylcholine as a modulator of neuronal differentiation has been considered in two experimental systems. A neuroblastoma cell line, which does not synthesise any neurotransmitter, has been transfected with a choline acetyltransferase construct; activation of acetylcholine synthesis, thus achieved, is followed by a higher expression of neuronal specific traits. The presence in these cells of muscarinic receptors is consistent with the existence of an autocrine loop, which may be responsible for the more advanced differentiation state observed in the transfected cells. Expression of cholinergic markers appears as a common featur...
Schwann cells (SC) are characterized by a remarkable plasticity that enables them to promptly res... more Schwann cells (SC) are characterized by a remarkable plasticity that enables them to promptly respond to nerve injury promoting axonal regeneration. In peripheral nerves after damage SC convert to a repair-promoting phenotype activating a sequence of supportive functions that drive myelin clearance, prevent neuronal death, and help axon growth and guidance. Regeneration of peripheral nerves after damage correlates inversely with thrombin levels. Thrombin is not only the key regulator of the coagulation cascade but also a protease with hormone-like activities that affects various cells of the central and peripheral nervous system mainly through the protease-activated receptor 1 (PAR1). Aim of the present study was to investigate if and how thrombin could affect the axon supportive functions of SC. In particular, our results show that the activation of PAR1 in rat SC cultures with low levels of thrombin or PAR1 agonist peptides induces the release of molecules, which favor neuronal su...
The bystander effect (BE) is an important biological phenomenon that induces damages in distant a... more The bystander effect (BE) is an important biological phenomenon that induces damages in distant and not directly affected by a chemical/physical stress cells. This effect, well known in ionizing radiation treatment, relies on reactive signals released by exposed cells and transmitted via cell–cell interaction or culture medium. In this study, cycloheximide (CHX)-induced apoptotic U937 cells and untreated THP-1 cells were chosen to investigate the chemical-induced BE. The effects of apoptotic U937 cells culture medium, Conditioned Medium (CM), on THP-1 cells were evaluated by morphological and immunohistochemical analysis performed by light microscopy; 1D 1H and 2D J-resolved (JRES) NMR metabolomic analysis has been used to characterize the molecules involved in the BE. In summary, this study indicates that: CM of CHX-treated U937 cells induces a time-dependent induction of toxicity, probably apoptotic cell death, and macrophagic differentiation in THP-1 cells; CM contains different ...
CHRNA7, encoding the neuronal alpha7 nicotinic acetylcholine receptor (a7nAChR), is highly expres... more CHRNA7, encoding the neuronal alpha7 nicotinic acetylcholine receptor (a7nAChR), is highly expressed in the brain, particularly in the hippocampus. It is situated in the 15q13.3 chromosome region, frequently associated with a Copy Number Variation (CNV), which causes its duplication or deletion. The clinical significance of CHRNA7 duplications is unknown so far, but there are several research data suggesting that they may be pathogenic, with reduced penetrance. We have produced an iPS cell line from a single healthy donor's fibroblasts carrying a 15q13.3 CNV, including CHRNA7 in order to study the exact role of this CNV during the neurodevelopment.
In previous studies, we found that the orthosteric muscarinic agonist arecaidine propargyl ester ... more In previous studies, we found that the orthosteric muscarinic agonist arecaidine propargyl ester (APE) (100 μM) induced a decreased cell proliferation and severe apoptosis in glioblastoma cancer stem cells (GSCs). In this report, we have investigated the effects mediated by hybrid (orthosteric/allosteric) muscarinic agonists P-6-Iper and N-8-Iper on GSCs survival. At variance with APE, the agonist N-8-Iper inhibited cell growth in a dose dependent manner and also impaired cell survival at low doses. The inhibitory effects of the N-8-Iper action appear to be mediated by M2 receptor activation, since they were strongly reduced by co-administration of the selective M2 receptor antagonist methoctramine as well as upon M2 receptor silencing. Moreover, analysis of the expression of phosphorylated histone H2AX (γ-H2AX) indicated that the treatment with N-8-Iper produced a decreased cell survival by induction of DNA damage. The ability of N-8-Iper to produce a cytotoxic effect and apoptosis...
Cns & Neurological Disorders-drug Targets, Oct 18, 2016
Exposure to environmental extremely low-frequency electromagnetic fields (ELF-EMF) in everyday li... more Exposure to environmental extremely low-frequency electromagnetic fields (ELF-EMF) in everyday life is increasing and it is a matter of great debate whether exposure to ELF-EMF can be harmful to human health. The neuropathology and symptoms of neurodegenerative disease depends on factors other than genetic predispositions, such as environmental exposure to disease-related risk factors. Research focusing on a possible contribution of ELF-EMF to cell injury and to the development of neurodegenerative disorders is characterized by conflicting data from epidemiological and animal studies. Due to lack of a direct link between neurodegenerative processes and ELF-EMF exposure, our goal was to investigate if ELF-EMF exposure may represent a possible risk factor. In the present study, using neuronal-like SH-SY5Y neuroblastoma cells, we show that the balance between generation and elimination of reactive oxygen species, as well as the balance between pro- and anti-inflammatory cytokines linked to oxidative stress, was maintained ensuring that cells respond properly to ELF-EMF (50Hz /1mT). In SH-SY5Y-exposed cells we observed increased intracellular 5-hydroxyindoleacetic acid/5-hydroxytryptamine ratio reflecting the rate of transmitter synthesis, catabolism and release, while matrix metalloproteinases that play critical roles in neuronal cell death were not significantly altered. The results presented here indicate that changes caused by short (1h-3h) and sub-chronic (48 h) exposure to 50Hz/1mT ELF-EMF in SH-SY5Y cells are minor in comparison to the neuronal cell damage expected to underlie neurodegeneration or cognitive impairment. Thus, these results are in accord with epidemiological studies that have provided little support for a link between ELF-EMFs and neurodegeneration.
Mesenchymal stem cells (MSCs), are multipotent cells present in several tissues and organs as bon... more Mesenchymal stem cells (MSCs), are multipotent cells present in several tissues and organs as bone marrow and adipose tissue. MSCs derived from Adipose tissue (ASCs) are an attractive source for regenerative medicine because they can be easily isolated, rapidly expandable in culture and show excellent differentiation potential. Acetylcholine (ACh), the most important neurotransmitter in central (CNS) and peripheral nervous system (PNS), plays a key role in the control of physiological processes also in non-neural tissues. Previously demonstrated that M2 muscarinic receptors negatively modulate ASC proliferation and migration. In the present work we demonstrate that rat ASCs also express \u3b17 nAChR receptors. By using the selective \u3b17 nicotinic receptor agonist ICH-3, we demonstrated that \u3b17 receptor activation causes a reduction of cell proliferation without affecting cell survival and morphology. Moreover, by wound healing assay, we have also demonstrated that \u3b17 promotes cell migration and a significant upregulation of CXCR4 receptor. Interestingly the activation of \u3b17 nAChR appears also up-regulate the expression of M2 acetylcholine receptor, suggesting a feedback positive loop between muscarinic and nicotinic receptors. Our data suggest that ACh, via cholinergic receptors might contribute to modulate ASC physiology. Moreover, in particular via nicotinic receptors, ACh might control ASCs migration
Glioblastomas are the most aggressive brain tumors in humans. They show a marked cell heterogenei... more Glioblastomas are the most aggressive brain tumors in humans. They show a marked cell heterogeneity; in particular glioblastoma stem cells (GCSs), a multipotent cell subpopulation capable of self-renewals, have been identified in these tumors. Considering that these cells show high resistance to chemiotherapy, the identification of drugs able to inhibit GCS proliferation and survival is considered of great clinical relevance. The involvement of muscarinic receptors in cancer has been reported. Recently we have demonstrated that the activation of M2 muscarinic receptors, through the agonist arecaidine, arrests cell proliferation and induces apoptosis in primary and stable glioblastoma cell lines. Following previous data, we have investigated the effect produced by M2 stimulation on GCS proliferation. The expression of M2 receptors was analyzed in several GCS cell lines derived from human biopsies. Considering the M2 receptor levels, we have selected two cell lines (GB7 and GB8). In both cell lines the treatment with the M2 agonist arecaidine decreases cell proliferation; in particular in GB7 cells the decrease of cell proliferation is time and dose dependent. Moreover co-treatment of GB7 cells with M1 and M3 antagonists (pirenzepine and 4-DAMP respectively) has indicated that the decreased cell proliferation\u2013arecaidine induced was probably dependent on M2 activation. FACS analysis has also shown that in GB7 cells, arecaidine inhibits cell cycle progression. Conversely in GB8 cells, arecaidine causes a decreased cell survival. The analysis of EGFR and Notch expression has clearly indicated that at least in GB7 cells the decrease of EGFR expression may be responsible on the decreased cell proliferation induced by arecaidine. Considering the previous data, the use of a more selective agonist for M2 receptor may have a strategic relevance for the glioblastoma therapy. As the difficulty of finding ligands with high selectivity, the exploitation of allosteric sites on G protein-coupled receptors (GPCRs) opens up a new range of possibilities in the research of selective orthosteric ligands in particular for GPCRs as mAChRs, which are characterized by highly conserved orthosteric sites among different receptor subtypes. Actually we are testing the effects produced by a new generation of dualsteric ligands for M2 receptors on glioblastoma cell proliferation both in stable cell lines and in the GCSs
Introduction. The involvement of muscarinic acetylcholine receptors (mAChRs) in cancer has been l... more Introduction. The involvement of muscarinic acetylcholine receptors (mAChRs) in cancer has been largely documented. Recently, we have demonstrated that activation of M2AChRs arrests cell proliferation and induces apoptosis in glioblastoma (GB) cell lines and in glioblastoma cancer stem cells (GSC). In the present work, we compared the effects mediated by the preferential activation of M2AChRs, through the M2 agonist arecaidine propargyl ester (APE) and the dualsteric agonist N8-Iper, both in GB cell lines and in GSC, analyzing their antiproliferative and pro-apoptotic effects and their potential use in combined therapy with conventional drugs (i.e., temozolomide). Material and Methods. U251 established cell lines and GSC (GB7 cell line) obtained from human biopsy were used as cell models. MTT assay, trypan blue staining and FACS analysis were used to evaluate cell viability and cell death. The M2 silencing (by siRNA) and pharmacological competition were used to confirm the ability of M2 agonists to selectively bind this receptor subtype. Transcript levels for multidrug efflux pumps (e.g., ATP binding cassette, ABC) were analyzed by Real-Time PCR and Western Blot analysis. Results. In GB7 cells, treatment with the N8-Iper decreased cell proliferation in a time and dose dependent manner as previously demonstrated after APE treatments. FACS analysis has also shown that in GB7 cells N8-Iper caused a significant increase of cell death. The pharmacological competition with M1 and M3 antagonists (pirenzepine and 4-DAMP, respectively) and the M2 antagonist methoctramine indicated that the effects of N8-Iper were dependent on selective activation of the M2AChR subtype. The ability of APE and N-8-Iper to activate M2 receptor populations was also confirmed by CHRM2 siRNA transfection. Moreover, APE and N-8-Iper decreased the mRNA levels and the protein expression of the ABC-G2 pump. Finally, the combined treatment with an M2 agonist and temozolomide indicated a synergic effect of the two drugs in decreasing GB cell survival. Conclusions. Our findings suggest that M2AChR agonists represent a new putative therapeutic tool in glioblastoma treatment. Moreover, the ability of M2 agonists to decrease the ABC-G2 drug efflux pumps expression in GSC cells, coupled with their synergic effect in combination with temozolomide, are indicative of their relevant role in reducing the GB chemoresistance
Neural tissue is a hierarchical multiscale system with intracellular and extracellular diffusion ... more Neural tissue is a hierarchical multiscale system with intracellular and extracellular diffusion compartments at different length scales. The normal diffusion of bulk water in tissues is not able to detect the specific features of a complex system, providing nonlocal, diffusion measurement averaged on a 10-20 μm length scale. Being able to probe tissues with sub-micrometric diffusion length and quantify new local parameters, transient anomalous diffusion (tAD) would dramatically increase the diagnostic potential of diffusion MRI (DMRI) in detecting collective and sub-micro architectural changes of human tissues due to pathological damage. In DMRI, the use of tAD parameters quantified using specific DMRI acquisition protocols and their interpretation has often aroused skepticism. Although the derived formulas may accurately fit experimental diffusion-weighted data, the relationships between the postulated dynamical feature and the underlying geometrical structure remains elusive, or ...
The International journal of developmental biology, 2000
Development of the nervous system is dependent on the co-operation between cell determination eve... more Development of the nervous system is dependent on the co-operation between cell determination events and the action of epigenetic factors; in addition to well known factors, e.g. growth factors, neurotransmitters have been assigned a role as "morphogens" and modulators of neuronal differentiation in an early developmental phase. The possible role of acetylcholine as a modulator of neuronal differentiation has been considered in two experimental systems. A neuroblastoma cell line, which does not synthesise any neurotransmitter, has been transfected with a choline acetyltransferase construct; activation of acetylcholine synthesis, thus achieved, is followed by a higher expression of neuronal specific traits. The presence in these cells of muscarinic receptors is consistent with the existence of an autocrine loop, which may be responsible for the more advanced differentiation state observed in the transfected cells. Expression of cholinergic markers appears as a common featur...
Schwann cells (SC) are characterized by a remarkable plasticity that enables them to promptly res... more Schwann cells (SC) are characterized by a remarkable plasticity that enables them to promptly respond to nerve injury promoting axonal regeneration. In peripheral nerves after damage SC convert to a repair-promoting phenotype activating a sequence of supportive functions that drive myelin clearance, prevent neuronal death, and help axon growth and guidance. Regeneration of peripheral nerves after damage correlates inversely with thrombin levels. Thrombin is not only the key regulator of the coagulation cascade but also a protease with hormone-like activities that affects various cells of the central and peripheral nervous system mainly through the protease-activated receptor 1 (PAR1). Aim of the present study was to investigate if and how thrombin could affect the axon supportive functions of SC. In particular, our results show that the activation of PAR1 in rat SC cultures with low levels of thrombin or PAR1 agonist peptides induces the release of molecules, which favor neuronal su...
The bystander effect (BE) is an important biological phenomenon that induces damages in distant a... more The bystander effect (BE) is an important biological phenomenon that induces damages in distant and not directly affected by a chemical/physical stress cells. This effect, well known in ionizing radiation treatment, relies on reactive signals released by exposed cells and transmitted via cell–cell interaction or culture medium. In this study, cycloheximide (CHX)-induced apoptotic U937 cells and untreated THP-1 cells were chosen to investigate the chemical-induced BE. The effects of apoptotic U937 cells culture medium, Conditioned Medium (CM), on THP-1 cells were evaluated by morphological and immunohistochemical analysis performed by light microscopy; 1D 1H and 2D J-resolved (JRES) NMR metabolomic analysis has been used to characterize the molecules involved in the BE. In summary, this study indicates that: CM of CHX-treated U937 cells induces a time-dependent induction of toxicity, probably apoptotic cell death, and macrophagic differentiation in THP-1 cells; CM contains different ...
CHRNA7, encoding the neuronal alpha7 nicotinic acetylcholine receptor (a7nAChR), is highly expres... more CHRNA7, encoding the neuronal alpha7 nicotinic acetylcholine receptor (a7nAChR), is highly expressed in the brain, particularly in the hippocampus. It is situated in the 15q13.3 chromosome region, frequently associated with a Copy Number Variation (CNV), which causes its duplication or deletion. The clinical significance of CHRNA7 duplications is unknown so far, but there are several research data suggesting that they may be pathogenic, with reduced penetrance. We have produced an iPS cell line from a single healthy donor's fibroblasts carrying a 15q13.3 CNV, including CHRNA7 in order to study the exact role of this CNV during the neurodevelopment.
In previous studies, we found that the orthosteric muscarinic agonist arecaidine propargyl ester ... more In previous studies, we found that the orthosteric muscarinic agonist arecaidine propargyl ester (APE) (100 μM) induced a decreased cell proliferation and severe apoptosis in glioblastoma cancer stem cells (GSCs). In this report, we have investigated the effects mediated by hybrid (orthosteric/allosteric) muscarinic agonists P-6-Iper and N-8-Iper on GSCs survival. At variance with APE, the agonist N-8-Iper inhibited cell growth in a dose dependent manner and also impaired cell survival at low doses. The inhibitory effects of the N-8-Iper action appear to be mediated by M2 receptor activation, since they were strongly reduced by co-administration of the selective M2 receptor antagonist methoctramine as well as upon M2 receptor silencing. Moreover, analysis of the expression of phosphorylated histone H2AX (γ-H2AX) indicated that the treatment with N-8-Iper produced a decreased cell survival by induction of DNA damage. The ability of N-8-Iper to produce a cytotoxic effect and apoptosis...
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Papers by Ada M Tata