Glioblastoma (GBM) is one of the most aggressive cancers and the most common primary brain tumor.... more Glioblastoma (GBM) is one of the most aggressive cancers and the most common primary brain tumor. Although the concomitant chemoradiotherapy using temozolomide (TMZ) has been established as the standard therapy, the median survival is approximately 14 months. Metformin is an oral biguanide introduced in the 1950s for the treatment of type 2 diabetes. A recent epidemiologic survey found that metformin also had significant effects on tumorigenesis. The purpose of the study is to investigate the efficacy of combined therapy with TMZ and metformin for the treatment of GBM in vitro and in vivo. Metformin and TMZ inhibit survival of U87MG cells in a dose-and time-dependent manner, respectively. The combination of metformin and temzolomide enhances the antitumor effect. Flow cytometry analysis showed that combination of metformin and temzolomide induces an increase in the percentage of apoptotic cells. Notably, the combination of metformin and TMZ enhances AMPK phosphorylation and inhibit mTOR, when compared with vehicle-treated cells and single-agent therapy. In xenograft models, the survival of mice treated with combined therapy with clinically relevant dosage of TMZ and metformin is similar with that of mice treated with single-agent therapy (67 vs. 56 days). Increased dosage of metformin combined with TMZ prolongs the survival in vivo injected with U87 or TMZ-resistance U87 cells. Our results show that metformin enhances the anti-tumor effect of TMZ via AMPK-mTOR signaling pathway. The presented data reinforce that metformin might be a good candidate for combined regimen with TMZ in clinical settings.
Positive or negative environmental modulations on human brain development: the morpho-functional outcomes of music training or stress
Frontiers in Neuroscience
In the last couple of decades, the study of human living brain has benefitted of neuroimaging and... more In the last couple of decades, the study of human living brain has benefitted of neuroimaging and non-invasive electrophysiological techniques, which are particularly valuable during development. A number of studies allowed to trace the usual stages leading from pregnancy to adult age, and relate them to functional and behavioral measurements. It was also possible to explore the effects of some interventions, behavioral or not, showing that the commonly followed pathway to adulthood may be steered by external interventions. These events may result in behavioral modifications but also in structural changes, in some cases limiting plasticity or extending/modifying critical periods. In this review, we outline the healthy human brain development in the absence of major issues or diseases. Then, the effects of negative (different stressors) and positive (music training) environmental stimuli on brain and behavioral development is depicted. Hence, it may be concluded that the typical deve...
Distribution of insoluble cAMP-dependent kinase type RI and RII in the lizard and turtle central nervous system
Brain Research, Jun 1, 2007
cAMP is a universal second messenger. In eucaryotes it acts mainly via protein kinases composed o... more cAMP is a universal second messenger. In eucaryotes it acts mainly via protein kinases composed of regulatory (R) and catalytic subunits; their subcellular distribution may differ according to the cell type. In rodent brain, peculiar detergent-insoluble RIalpha aggregates were previously described in neurons of areas related to the limbic system, while RIIbeta is more evenly distributed also in non-nervous cells. It is unclear whether the regional distribution of regulatory subunits is typical of mammalian brain. Western blots and immunohistochemistry showed that in lizard brains a large fraction of the cAMP-dependent protein kinase regulatory isoforms is insoluble, as in mammals. Insoluble RIalpha and RII regulatory isoforms were not evenly distributed but organized in clearly separated aggregates. Numerous RII aggregates were present in almost all brain regions and were found also in non-nervous cells. As shown by immunohistochemistry and equilibrium binding of fluorescently tagged cAMP, RIalpha aggregates were restricted to neurons of some brain regions: telencephalon, particularly medial cortical areas, dorsal ventricular ridge, olfactory pathways, medial hypothalamus and cerebellar granular layer were intensely labelled. A very weak RIalpha labelling was detected in the brainstem reticular formation, in the periaqueductal gray and in the spinal cord dorsal horn. A similar distribution of RIalpha aggregates was also found in turtle brains. Their distribution is reminiscent of that observed in mammals, although with some differences in relative intensity and persistence. The supramolecular organization of the RIalpha isoform may help in establishing homologies and differences between brain areas involved in visceroemotional control.
Vomeronasal Receptors and Signal Transduction in the Vomeronasal Organ of Mammals -- Neurobiology of Chemical Communication
Intraspecific communication involves the activation of chemoreceptors and subsequent activation o... more Intraspecific communication involves the activation of chemoreceptors and subsequent activation of different central areas that coordinate the responses of the entire organism\u2014ranging from behavioral modification to modulation of hormones release. Animals emit intraspecific chemical signals, often referred to as pheromones, to advertise their presence to members of the same species and to regulate interactions aimed at establishing and regulating social and reproductive bonds. In the last two decades, scientists have developed a greater understanding of the neural processing of these chemical signals. Neurobiology of Chemical Communication explores the role of the chemical senses in mediating intraspecific communication. Providing an up-to-date outline of the most recent advances in the field, it presents data from laboratory and wild species, ranging from invertebrates to vertebrates, from insects to humans. The book examines the structure, anatomy, electrophysiology, and molecular biology of pheromones. It discusses how chemical signals work on different mammalian and non-mammalian species and includes chapters on insects, Drosophila, honey bees, amphibians, mice, tigers, and cattle. It also explores the controversial topic of human pheromones. An essential reference for students and researchers in the field of pheromones, this is also an ideal resource for those working on behavioral phenotyping of animal models and persons interested in the biology/ecology of wild and domestic species
THE expression of selected regulatory neuropeptides was investigated by immunohistochemistry in n... more THE expression of selected regulatory neuropeptides was investigated by immunohistochemistry in nerves supplying the vomeronasal organ (VNO) of mice during postnatal development. Results show that neurons in the VNO are devoid of immunolabeling with any of the antibody used from 1 day to 2 months of age. In the non-receptor epithelium (NRE) and the vomeronasal vascular pump (VP) the timing of expression of regulatory neuropeptides differed among neuropeptides and the different VNO structures. Regulatory neuropeptides usually found in sensory nerves (substance P, calcitonin gene-related peptide) and efferent nerves (neuropeptide Y, atrial natriuretic peptide) are expressed in the NRE and the VP, respectively. These results support the view that the VNO is to some extent functional during postnatal development. Neuro-Report 10:2023±2027 # 1999 Lippincott Williams & Wilkins.
Molecular and Neural Mechanisms of Pheromone Reception in the Rat Vomeronasal System and Changes in the Pheromonal Reception by the Maturation and Sexual Experiences
Knockout mice lacking the sweet taste receptor subunit Tas1r3 lose behavioral and nerve responses... more Knockout mice lacking the sweet taste receptor subunit Tas1r3 lose behavioral and nerve responses to non-caloric sweeteners but retain responses to glucose and other sugars. This suggests the presence of a Tas1r3-independent mechanism to detect the sweetness of sugars. We used PCR, in situ hybridization and immunohistochemistry to identify signal transduction elements selectively expressed in sweet-responsive taste cells that might mediate Tas1r3independent responses to sugars. Glucose transporters (GLUTs), Na-dependent glucose co-transporter-1 (SGLT1) and ATP-gated K + channels (K ATP) are all present in Tas1r3+ taste cells and may constitute a Tas1r3independent means to detect caloric sugars. In addition, Tas1r3+ taste cells selectively express disaccharidase enzymes sucrase, maltase, trehalase and lactase. In both wildtype and Tas1r3 knockout mice disaccharidase inhibitors significantly reduced gustatory nerve responses to the disaccharides sucrose and maltose, but not to the monosaccharides glucose and fructose or the noncaloric sweeteners. It appears that these orally expressed enzymes act in concert with salivary amylase to generate free glucose from starch, sucrose, maltose, trehalose and lactose that can activate the Tas1r3-independent sugar detection pathway. To identify additional signalling components and regulatory factors selectively expressed in Tas1r3+ taste cells we used single taste cell RNA-Seq (deep sequencing) and bioinformatics to "data-mine" the Tas1r3+ taste cell "transcriptome" (i.e. all genes transcribed in Tas1r3+ taste cells). Gli3, a key transcriptional effector in the sonic hedgehog signalling pathway, was found to be selectively expressed in Tas1r3+ taste cells and Lgr-5+ taste stem cells, but not in type I or type III taste cells. Gli3 conditional knockout mice were more sensitive to sweet, umami and bitter compounds and had increased numbers of type II taste cells. Our results suggest that Gli3 is an important negative regulator of taste bud maintenance that enhances the number of type II taste cells, including those responsive to sweet.
Deregulation of intracellular signal transduction pathways is a hallmark of cancer cells, clearly... more Deregulation of intracellular signal transduction pathways is a hallmark of cancer cells, clearly differentiating them from healthy cells. Differential intracellular distribution of the cAMPdependent protein kinases (PKA) was previously detected in cell cultures and in vivo in glioblastoma and medulloblastoma. Our goal is to extend this observation to meningioma, to explore possible differences among tumors of different origins and prospective outcomes. The distribution of regulatory and catalytic subunits of PKA has been examined in tissue specimens obtained during surgery from meningioma patients. PKA RI subunit appeared more evenly distributed throughout the cytoplasm, but it was clearly detectable only in some tumors. RII was present in discrete spots, presumably at high local concentration; these aggregates could also be visualized under equilibrium binding conditions with fluorescent 8-substituted cAMP analogues, at variance with normal brain tissue and other brain tumors. The PKA catalytic subunit showed exactly overlapping pattern to RII and in fixed sections could be visualized by fluorescent cAMP analogues. Gene expression analysis showed that the PKA catalytic subunit revealed a significant correlation pattern with genes involved in meningioma. Hence, meningioma patients show a distinctive distribution pattern of PKA regulatory and catalytic subunits, different from glioblastoma, medulloblastoma, and healthy brain tissue. These observations raise the possibility of exploiting the PKA intracellular pathway as a diagnostic tool and possible therapeutic interventions.
American Journal of Physiology-lung Cellular and Molecular Physiology, Dec 1, 2009
Asthma needs continuous treatment often for years. In humans, some drugs are administered via aer... more Asthma needs continuous treatment often for years. In humans, some drugs are administered via aerosol, therefore they come in contact with both respiratory and olfactory mucosa. We explored the possibility that antiasthma corticosteroid treatment could influence the olfactory function by passage through the nose. A group of mice was exposed twice daily for 42 days to fluticasone propionate aerosol and was compared with a control group. Olfactory behavior, respiratory mechanics, histology, and immunoreactivity in the olfactory system were assessed. Fluticasone-treated mice were slower in retrieving a piece of hidden food, but both groups were similarly fast when the food was visible. When a clearly detectable odor was present in the environment, all mice behaved in a similar way. Respiratory mechanics indices were similar in all mice except for the viscose resistance, which was reduced in fluticasone-treated mice. Olfactory mucosa of fluticasone-treated mice was thicker than that of controls. Slight but consistent differences in staining were present for Olfactory Marker Protein but not for other proteins. A mild impairment of olfactory function is present in mice chronically treated with fluticasone aerosol, apparently accompanied by slight modifications of the olfactory receptor cells, and suggests monitoring of olfactory function modifications in long-term steroid users.
Identification of alpha2u-globulin and bound volatiles in the Indian common house rat (Rattus rattus)
PubMed, Aug 1, 2009
The alpha2u-globulin (alpha2u) is a pheromone carrier urinary protein believed to be relevant for... more The alpha2u-globulin (alpha2u) is a pheromone carrier urinary protein believed to be relevant for sexual communication among rats and is characterized in laboratory rats. In the present study 17 kDa protein and the bound pheromones were characterized in a population of wild-type Indian common house rat (Rattus rattus). The protein was purified by two runs of Sephadex G-50 chromatography and analyzed with SDS-PAGE with MALDI-TOF/MS. The results of MASCOT search identified the protein as an alpha2u and suggested a role for binding pheromones. To confirm the protein bound volatiles, purified alpha2u was extracted with dichloromethane and volatile molecules were detected using of gas chromatography linked to mass spectrometry (GC-MS). 1-Chlorodecane was detected as the predominant compound and 2-methyl-N-phenyl-2-propenamide, hexadecane and 2,6,11-trimethyl decane as the minor compounds. The simple method of protein purification and the identification of bound volatiles may help in designing efficient pheromone-based rat traps.
Glioblastoma (GBM) has the worst prognosis among brain tumors, hence basic biology, preclinical, ... more Glioblastoma (GBM) has the worst prognosis among brain tumors, hence basic biology, preclinical, and clinical studies are necessary to design effective strategies to defeat this disease. Gene transfer vectors derived from the most-studied lentivirus-the Human Immunodeficiency Virus type 1-have wide application in dissecting GBM specific features to identify potential therapeutic targets. Last-generation lentiviruses (LV), highly improved in safety profile and gene transfer capacity, are also largely employed as delivery systems of therapeutic molecules to be employed in gene therapy (GT) approaches. LV were initially used in GT protocols aimed at the expression of suicide factors to induce GBM cell death. Subsequently, LV were adopted to either express small noncoding RNAs to affect different aspects of GBM biology or to overcome the resistance to both chemo-and radiotherapy that easily develop in this tumor after initial therapy. Newer frontiers include adoption of LV for engineering T cells to express chimeric antigen receptors recognizing specific GBM antigens, or for transducing specific cell types that, due to their biological properties, can function as carriers of therapeutic molecules to the cancer mass. Finally, LV allow the setting up of improved animal models crucial for the validation of GBM specific therapies.
Background: Olfaction is often affected in parkinsonian patients, but dopaminergic cells in the o... more Background: Olfaction is often affected in parkinsonian patients, but dopaminergic cells in the olfactory bulb are not affected by some Parkinson-inducing drugs. We investigated whether the drug MPTP produces the olfactory deficits typical of Parkinson and affects the olfactory bulb in mice. Findings: Lesioned and control mice were tested for olfactory search, for motor and exploratory behavior. Brains and olfactory mucosa were investigated via immunohistochemistry for thyrosine hydroxylase, Olfactory Marker Protein and cyclic AMP-dependent protein kinase as an intracellular pathway involved in dopaminergic neurotransmission. MPTP induced motor impairment, but no deficit in olfactory search. Thyrosine hydroxylase did not differ in olfactory bulb, while a strong decrease was detected in substantia nigra and tegmentum of MPTP mice. Olfactory Marker Protein decreased in the olfactory bulb of MPTP mice, while a cyclic AMP-dependent protein kinase increased in the inner granular layer of MPTP mice. Conclusions: MPTP mice do not present behavioural deficits in olfactory search, yet immunoreactivity reveals modifications in the olfactory bulb, and suggests changes in intracellular signal processing, possibly linked to neuron survival after MPTP.
Glioblastoma (GBM) is one of the most aggressive cancers and the most common primary brain tumor.... more Glioblastoma (GBM) is one of the most aggressive cancers and the most common primary brain tumor. Although the concomitant chemoradiotherapy using temozolomide (TMZ) has been established as the standard therapy, the median survival is approximately 14 months. Metformin is an oral biguanide introduced in the 1950s for the treatment of type 2 diabetes. A recent epidemiologic survey found that metformin also had significant effects on tumorigenesis. The purpose of the study is to investigate the efficacy of combined therapy with TMZ and metformin for the treatment of GBM in vitro and in vivo. Metformin and TMZ inhibit survival of U87MG cells in a dose-and time-dependent manner, respectively. The combination of metformin and temzolomide enhances the antitumor effect. Flow cytometry analysis showed that combination of metformin and temzolomide induces an increase in the percentage of apoptotic cells. Notably, the combination of metformin and TMZ enhances AMPK phosphorylation and inhibit mTOR, when compared with vehicle-treated cells and single-agent therapy. In xenograft models, the survival of mice treated with combined therapy with clinically relevant dosage of TMZ and metformin is similar with that of mice treated with single-agent therapy (67 vs. 56 days). Increased dosage of metformin combined with TMZ prolongs the survival in vivo injected with U87 or TMZ-resistance U87 cells. Our results show that metformin enhances the anti-tumor effect of TMZ via AMPK-mTOR signaling pathway. The presented data reinforce that metformin might be a good candidate for combined regimen with TMZ in clinical settings.
Positive or negative environmental modulations on human brain development: the morpho-functional outcomes of music training or stress
Frontiers in Neuroscience
In the last couple of decades, the study of human living brain has benefitted of neuroimaging and... more In the last couple of decades, the study of human living brain has benefitted of neuroimaging and non-invasive electrophysiological techniques, which are particularly valuable during development. A number of studies allowed to trace the usual stages leading from pregnancy to adult age, and relate them to functional and behavioral measurements. It was also possible to explore the effects of some interventions, behavioral or not, showing that the commonly followed pathway to adulthood may be steered by external interventions. These events may result in behavioral modifications but also in structural changes, in some cases limiting plasticity or extending/modifying critical periods. In this review, we outline the healthy human brain development in the absence of major issues or diseases. Then, the effects of negative (different stressors) and positive (music training) environmental stimuli on brain and behavioral development is depicted. Hence, it may be concluded that the typical deve...
Distribution of insoluble cAMP-dependent kinase type RI and RII in the lizard and turtle central nervous system
Brain Research, Jun 1, 2007
cAMP is a universal second messenger. In eucaryotes it acts mainly via protein kinases composed o... more cAMP is a universal second messenger. In eucaryotes it acts mainly via protein kinases composed of regulatory (R) and catalytic subunits; their subcellular distribution may differ according to the cell type. In rodent brain, peculiar detergent-insoluble RIalpha aggregates were previously described in neurons of areas related to the limbic system, while RIIbeta is more evenly distributed also in non-nervous cells. It is unclear whether the regional distribution of regulatory subunits is typical of mammalian brain. Western blots and immunohistochemistry showed that in lizard brains a large fraction of the cAMP-dependent protein kinase regulatory isoforms is insoluble, as in mammals. Insoluble RIalpha and RII regulatory isoforms were not evenly distributed but organized in clearly separated aggregates. Numerous RII aggregates were present in almost all brain regions and were found also in non-nervous cells. As shown by immunohistochemistry and equilibrium binding of fluorescently tagged cAMP, RIalpha aggregates were restricted to neurons of some brain regions: telencephalon, particularly medial cortical areas, dorsal ventricular ridge, olfactory pathways, medial hypothalamus and cerebellar granular layer were intensely labelled. A very weak RIalpha labelling was detected in the brainstem reticular formation, in the periaqueductal gray and in the spinal cord dorsal horn. A similar distribution of RIalpha aggregates was also found in turtle brains. Their distribution is reminiscent of that observed in mammals, although with some differences in relative intensity and persistence. The supramolecular organization of the RIalpha isoform may help in establishing homologies and differences between brain areas involved in visceroemotional control.
Vomeronasal Receptors and Signal Transduction in the Vomeronasal Organ of Mammals -- Neurobiology of Chemical Communication
Intraspecific communication involves the activation of chemoreceptors and subsequent activation o... more Intraspecific communication involves the activation of chemoreceptors and subsequent activation of different central areas that coordinate the responses of the entire organism\u2014ranging from behavioral modification to modulation of hormones release. Animals emit intraspecific chemical signals, often referred to as pheromones, to advertise their presence to members of the same species and to regulate interactions aimed at establishing and regulating social and reproductive bonds. In the last two decades, scientists have developed a greater understanding of the neural processing of these chemical signals. Neurobiology of Chemical Communication explores the role of the chemical senses in mediating intraspecific communication. Providing an up-to-date outline of the most recent advances in the field, it presents data from laboratory and wild species, ranging from invertebrates to vertebrates, from insects to humans. The book examines the structure, anatomy, electrophysiology, and molecular biology of pheromones. It discusses how chemical signals work on different mammalian and non-mammalian species and includes chapters on insects, Drosophila, honey bees, amphibians, mice, tigers, and cattle. It also explores the controversial topic of human pheromones. An essential reference for students and researchers in the field of pheromones, this is also an ideal resource for those working on behavioral phenotyping of animal models and persons interested in the biology/ecology of wild and domestic species
THE expression of selected regulatory neuropeptides was investigated by immunohistochemistry in n... more THE expression of selected regulatory neuropeptides was investigated by immunohistochemistry in nerves supplying the vomeronasal organ (VNO) of mice during postnatal development. Results show that neurons in the VNO are devoid of immunolabeling with any of the antibody used from 1 day to 2 months of age. In the non-receptor epithelium (NRE) and the vomeronasal vascular pump (VP) the timing of expression of regulatory neuropeptides differed among neuropeptides and the different VNO structures. Regulatory neuropeptides usually found in sensory nerves (substance P, calcitonin gene-related peptide) and efferent nerves (neuropeptide Y, atrial natriuretic peptide) are expressed in the NRE and the VP, respectively. These results support the view that the VNO is to some extent functional during postnatal development. Neuro-Report 10:2023±2027 # 1999 Lippincott Williams & Wilkins.
Molecular and Neural Mechanisms of Pheromone Reception in the Rat Vomeronasal System and Changes in the Pheromonal Reception by the Maturation and Sexual Experiences
Knockout mice lacking the sweet taste receptor subunit Tas1r3 lose behavioral and nerve responses... more Knockout mice lacking the sweet taste receptor subunit Tas1r3 lose behavioral and nerve responses to non-caloric sweeteners but retain responses to glucose and other sugars. This suggests the presence of a Tas1r3-independent mechanism to detect the sweetness of sugars. We used PCR, in situ hybridization and immunohistochemistry to identify signal transduction elements selectively expressed in sweet-responsive taste cells that might mediate Tas1r3independent responses to sugars. Glucose transporters (GLUTs), Na-dependent glucose co-transporter-1 (SGLT1) and ATP-gated K + channels (K ATP) are all present in Tas1r3+ taste cells and may constitute a Tas1r3independent means to detect caloric sugars. In addition, Tas1r3+ taste cells selectively express disaccharidase enzymes sucrase, maltase, trehalase and lactase. In both wildtype and Tas1r3 knockout mice disaccharidase inhibitors significantly reduced gustatory nerve responses to the disaccharides sucrose and maltose, but not to the monosaccharides glucose and fructose or the noncaloric sweeteners. It appears that these orally expressed enzymes act in concert with salivary amylase to generate free glucose from starch, sucrose, maltose, trehalose and lactose that can activate the Tas1r3-independent sugar detection pathway. To identify additional signalling components and regulatory factors selectively expressed in Tas1r3+ taste cells we used single taste cell RNA-Seq (deep sequencing) and bioinformatics to "data-mine" the Tas1r3+ taste cell "transcriptome" (i.e. all genes transcribed in Tas1r3+ taste cells). Gli3, a key transcriptional effector in the sonic hedgehog signalling pathway, was found to be selectively expressed in Tas1r3+ taste cells and Lgr-5+ taste stem cells, but not in type I or type III taste cells. Gli3 conditional knockout mice were more sensitive to sweet, umami and bitter compounds and had increased numbers of type II taste cells. Our results suggest that Gli3 is an important negative regulator of taste bud maintenance that enhances the number of type II taste cells, including those responsive to sweet.
Deregulation of intracellular signal transduction pathways is a hallmark of cancer cells, clearly... more Deregulation of intracellular signal transduction pathways is a hallmark of cancer cells, clearly differentiating them from healthy cells. Differential intracellular distribution of the cAMPdependent protein kinases (PKA) was previously detected in cell cultures and in vivo in glioblastoma and medulloblastoma. Our goal is to extend this observation to meningioma, to explore possible differences among tumors of different origins and prospective outcomes. The distribution of regulatory and catalytic subunits of PKA has been examined in tissue specimens obtained during surgery from meningioma patients. PKA RI subunit appeared more evenly distributed throughout the cytoplasm, but it was clearly detectable only in some tumors. RII was present in discrete spots, presumably at high local concentration; these aggregates could also be visualized under equilibrium binding conditions with fluorescent 8-substituted cAMP analogues, at variance with normal brain tissue and other brain tumors. The PKA catalytic subunit showed exactly overlapping pattern to RII and in fixed sections could be visualized by fluorescent cAMP analogues. Gene expression analysis showed that the PKA catalytic subunit revealed a significant correlation pattern with genes involved in meningioma. Hence, meningioma patients show a distinctive distribution pattern of PKA regulatory and catalytic subunits, different from glioblastoma, medulloblastoma, and healthy brain tissue. These observations raise the possibility of exploiting the PKA intracellular pathway as a diagnostic tool and possible therapeutic interventions.
American Journal of Physiology-lung Cellular and Molecular Physiology, Dec 1, 2009
Asthma needs continuous treatment often for years. In humans, some drugs are administered via aer... more Asthma needs continuous treatment often for years. In humans, some drugs are administered via aerosol, therefore they come in contact with both respiratory and olfactory mucosa. We explored the possibility that antiasthma corticosteroid treatment could influence the olfactory function by passage through the nose. A group of mice was exposed twice daily for 42 days to fluticasone propionate aerosol and was compared with a control group. Olfactory behavior, respiratory mechanics, histology, and immunoreactivity in the olfactory system were assessed. Fluticasone-treated mice were slower in retrieving a piece of hidden food, but both groups were similarly fast when the food was visible. When a clearly detectable odor was present in the environment, all mice behaved in a similar way. Respiratory mechanics indices were similar in all mice except for the viscose resistance, which was reduced in fluticasone-treated mice. Olfactory mucosa of fluticasone-treated mice was thicker than that of controls. Slight but consistent differences in staining were present for Olfactory Marker Protein but not for other proteins. A mild impairment of olfactory function is present in mice chronically treated with fluticasone aerosol, apparently accompanied by slight modifications of the olfactory receptor cells, and suggests monitoring of olfactory function modifications in long-term steroid users.
Identification of alpha2u-globulin and bound volatiles in the Indian common house rat (Rattus rattus)
PubMed, Aug 1, 2009
The alpha2u-globulin (alpha2u) is a pheromone carrier urinary protein believed to be relevant for... more The alpha2u-globulin (alpha2u) is a pheromone carrier urinary protein believed to be relevant for sexual communication among rats and is characterized in laboratory rats. In the present study 17 kDa protein and the bound pheromones were characterized in a population of wild-type Indian common house rat (Rattus rattus). The protein was purified by two runs of Sephadex G-50 chromatography and analyzed with SDS-PAGE with MALDI-TOF/MS. The results of MASCOT search identified the protein as an alpha2u and suggested a role for binding pheromones. To confirm the protein bound volatiles, purified alpha2u was extracted with dichloromethane and volatile molecules were detected using of gas chromatography linked to mass spectrometry (GC-MS). 1-Chlorodecane was detected as the predominant compound and 2-methyl-N-phenyl-2-propenamide, hexadecane and 2,6,11-trimethyl decane as the minor compounds. The simple method of protein purification and the identification of bound volatiles may help in designing efficient pheromone-based rat traps.
Glioblastoma (GBM) has the worst prognosis among brain tumors, hence basic biology, preclinical, ... more Glioblastoma (GBM) has the worst prognosis among brain tumors, hence basic biology, preclinical, and clinical studies are necessary to design effective strategies to defeat this disease. Gene transfer vectors derived from the most-studied lentivirus-the Human Immunodeficiency Virus type 1-have wide application in dissecting GBM specific features to identify potential therapeutic targets. Last-generation lentiviruses (LV), highly improved in safety profile and gene transfer capacity, are also largely employed as delivery systems of therapeutic molecules to be employed in gene therapy (GT) approaches. LV were initially used in GT protocols aimed at the expression of suicide factors to induce GBM cell death. Subsequently, LV were adopted to either express small noncoding RNAs to affect different aspects of GBM biology or to overcome the resistance to both chemo-and radiotherapy that easily develop in this tumor after initial therapy. Newer frontiers include adoption of LV for engineering T cells to express chimeric antigen receptors recognizing specific GBM antigens, or for transducing specific cell types that, due to their biological properties, can function as carriers of therapeutic molecules to the cancer mass. Finally, LV allow the setting up of improved animal models crucial for the validation of GBM specific therapies.
Background: Olfaction is often affected in parkinsonian patients, but dopaminergic cells in the o... more Background: Olfaction is often affected in parkinsonian patients, but dopaminergic cells in the olfactory bulb are not affected by some Parkinson-inducing drugs. We investigated whether the drug MPTP produces the olfactory deficits typical of Parkinson and affects the olfactory bulb in mice. Findings: Lesioned and control mice were tested for olfactory search, for motor and exploratory behavior. Brains and olfactory mucosa were investigated via immunohistochemistry for thyrosine hydroxylase, Olfactory Marker Protein and cyclic AMP-dependent protein kinase as an intracellular pathway involved in dopaminergic neurotransmission. MPTP induced motor impairment, but no deficit in olfactory search. Thyrosine hydroxylase did not differ in olfactory bulb, while a strong decrease was detected in substantia nigra and tegmentum of MPTP mice. Olfactory Marker Protein decreased in the olfactory bulb of MPTP mice, while a cyclic AMP-dependent protein kinase increased in the inner granular layer of MPTP mice. Conclusions: MPTP mice do not present behavioural deficits in olfactory search, yet immunoreactivity reveals modifications in the olfactory bulb, and suggests changes in intracellular signal processing, possibly linked to neuron survival after MPTP.
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Papers by Carla Mucignat