Mariarosaria Miloso | Università degli Studi di Milano-Bicocca (original) (raw)

Papers by Mariarosaria Miloso

Molecules, 2017

The Editorial Board of the Medicinal Chemistry section of the journal Molecules publishes here it... more The Editorial Board of the Medicinal Chemistry section of the journal Molecules publishes here its first Editorial, which has been prepared by highlighting, in sub-editorials of about one hundred words, some selected recently published articles that may have a profound impact on drug discovery and therapy.[...]

Journal of Bacteriology, 1993

We have cloned and sequenced the genomic regions encompassing the rho genes of Neisseria gonorrho... more We have cloned and sequenced the genomic regions encompassing the rho genes of Neisseria gonorrhoeae and Salmonella typhimurium. Rho factor of S. typhimurium has only three amino acid differences with respect to the Escherichia coli homolog. Northern (RNA) blots and primer extension experiments were used to characterize the N. gonorrhoeae rho transcript and to identify the transcription initiation and termination elements of this cistron. The function of the Rho factor of N. gonorrhoeae was investigated by complementation assays of rho mutants of E. coli and S. typhimurium and by in vivo transcription assays in polar mutants of S. typhimurium.

The new microbiologica, 1994

Molecular genetics appears to be the most promising approach to understanding the biology and pat... more Molecular genetics appears to be the most promising approach to understanding the biology and pathology of Chlamydia. This report focuses on the cloning and the protein expression of a DNA fragment from Chlamydia trachomatis DK20 chromosome. Results of hybridization experiments suggest that this sequence is specifically present within chlamydial DNA. The coding capacity of this DNA fragment is supported by the expression of a 26,000 m.w. peptide, in an Escherichia coli maxicell system.

Giornale di batteriologia, virologia ed immunologia

In this study the MicroTrak (Syva Co.) system has been evaluated in comparison with the isolation... more In this study the MicroTrak (Syva Co.) system has been evaluated in comparison with the isolation of Chlamydia trachomatis onto McCoy cell monolayer, testing 120 endocervical samples taken from an unselected women population. The overall results show a higher incidence of positivity in women under 30 years of age; a sensitivity of the immunofluorescence of 76.3% and a specificity of 89.8%. The agreement between the two methods is significative when we test symptomatic patients.

Giornale di batteriologia, virologia ed immunologia

The Authors have evaluated, by DNA-probe hybridization, the presence of CMV in the cellular fract... more The Authors have evaluated, by DNA-probe hybridization, the presence of CMV in the cellular fractions of different body fluids from immunocompromised patients. This specific methodology appear to be rapid to perform, adaptable to different clinical samples and sensitive. Even in absence of a supporting evaluation of specific antibodies, direct detection of virus, by DNA-probe, seems to be the most efficient and rapid approach to confirm the diagnosis either of active phase or reactivating of CMV infection.

International Journal of Molecular Sciences

Rigosertib is a small molecule in preclinical development that, due to its characteristics as a d... more Rigosertib is a small molecule in preclinical development that, due to its characteristics as a dual PLK1 and PI3K inhibitor, is particularly effective in counteracting the advance of different types of tumors. In this work, we evaluated the efficacy of Rigosertib and the expression of p53 in five different human tumor cell lines in vitro, A549 (lung adenocarcinoma), MCF-7 and MDA-MB231 (breast cancer cells), RPMI 8226 (multiple myeloma), and U87-MG (glioblastoma). We demonstrated that in all cell lines, the effect was dose- and time-dependent, but A549 cells were the most sensible to the treatment while higher concentrations were required for the most resistant cell line U87-MG. Moreover, the highest and lowest p53 levels have been observed, respectively, in A459 and U87-MG cells. The alterations in the cell cycle and in cell-cycle-related proteins were observed in A549 at lower concentrations than U87-MG. In conclusion, with this article we have demonstrated that Rigosertib has di...

Italian journal of anatomy and embryology, 2015

Oxygen and glucose deprivation (OGD) due to ischemic events or trauma in the brain result in neur... more Oxygen and glucose deprivation (OGD) due to ischemic events or trauma in the brain result in neuronal loss. The therapeutic approaches available inadequate and often the outcome is unfavorable for the patient or at least unpredictable. Stem cells could be useful for the treatment of OGD injured-neurons. Mesenchymal Stem Cells (MSCs), isolated from bone marrow as well as from various tissues, have poor immunogenicity and neuroprotective properties being able to alleviate ischemic brain injuries in animal models. The Endothelial Progenitor Cells (EPCs) are present at low frequencies both in the bone marrow and in the peripheral blood. They are thought to play a role in the recovery of cerebrovasculature integrity after stroke. In the present study we evaluated the potential neuroprotective effect of human MSCs and human EPCs on rat embryonic cortical neurons injured by OGD. OGD was induced by incubating the cortical neurons in a hypoxia chamber in a 95% N2 + 5% CO2 atmosphere at 37°C ...

Italian journal of anatomy and embryology = Archivio italiano di anatomia ed embriologia, 2012

The peripheral nervous system is a frequent target of toxic agents. The accurate identification o... more The peripheral nervous system is a frequent target of toxic agents. The accurate identification of the sites of neurotoxic action through the morphological characterization of reliable in vivo models or in vitro systems can give fundamental clues when investigating the pathogenesis and interpreting the clinical features of drug-induced neuropathy. The morphological approach has been used to investigate almost all the anticancer drugs able to induce chemotherapy-induced peripheral neurotoxicity, i.e. platinum drugs, antitubulins and proteasome inhibitors. No models have ever been described for thalidomide. This review demonstrates that any pathogenetic study on chemotherapy-induced peripheral neurotoxicity must be based on solid morphological observations obtained in reliable animal and in vitro models. This is particularly true in this setting, since the availability of tissues of human origin is extremely limited. In fact, peripheral (generally sural) nerve biopsies are never requi...

Hibiscus Sabdariffa (HS) is a plant commonly used in folk medicine (1). In recent years HS has ga... more Hibiscus Sabdariffa (HS) is a plant commonly used in folk medicine (1). In recent years HS has gained great interest due to its important antioxidant, anti-inflammatory and antitumoral properties. In our work, we evaluated the in vitro anticancer effects of HS extract against two different human breast cancer cell lines: estrogen receptor (ER) positive MCF-7 cells and ER negative MDA-MB-231 cells. We tested both total extract (HSE) and one fraction obtained by ethyl acetate extraction (HSEC). MTT assay and Trypan Blue vital count showed a dose and time dependent reduction of the viability in both cell lines treated with different concentrations of HSE or HSEC compared to untreated control cells. A significantly marked reduction was observed in MCF-7 cells treated with HSEC. On the basis of our results we used the concentrations of 7.5mg/ml and 3.5mg/ml respectively for HSE and HSEC. In order to evaluate ER involvement in HS effect, we analyzed the cellular localization of the recept...

The quantum-mechanical effects typical for single atoms or molecules can be reproduced in microme... more The quantum-mechanical effects typical for single atoms or molecules can be reproduced in micrometer-scale electric devices. In these systems the essential component is a small Josephson junction (JJ) consisting of two superconductors separated by a thin insulator. The quantum phenomena can be controlled in real time by external signals and have a great potential for novel applications. However, their fragility on uncontrolled disturbance caused by typical nearby environments is a drawback for quantum information science, but a virtue for detector technology. Motivated by this we have theoretically studied transistor kind of devices based on single-charge tunneling through small JJs. A common factor of the research is the analysis of the interplay between the coherent Cooperpair (charge carriers in the superconducting state) tunneling and incoherent environmental processes. In the first work we calculate the current due to incoherent Cooper-pair tunneling through a voltage-biased small JJ in series with large JJs and compare the results with recent experiments. We are able to reproduce the main experimental features and interpret these as traces of energy levels and energy bands of the mesoscopic device. In the second work we analyze a similar circuit (asymmetric single-Cooper-pair transistor) but under the assumption that the Cooper-pair tunneling is mainly coherent. This predicts new resonant transport voltages in the circuit due to higher-order processes. However, no clear traces of most of them are seen in the experiments, and similar discrepancy is present also in the case of the symmetric circuit. We continue to study this problem by modeling the interplay between the coherent and incoherent processes more accurately using a density-matrix approach. By this we are able to demonstrate that in typical conditions most of these resonances are indeed washed out by strong decoherence caused by the environment. We also analyze the contribution of three typical weakly interacting dissipative environments: electromagnetic environment, spurious charge fluctuators in the nearby insulating materials, and quasiparticles. In the last work we model the dynamics of a currentbiased JJ perturbed by a smaller JJ using a similar density-matrix approach. We demonstrate that the small JJ can be used also as a detector of the energy-band dynamics in a current biased JJ. The method is also used for modeling the charge transport in the Bloch-oscillating transistor.

Italian journal of anatomy and embryology, 2011

Nowadays obesity and its related diseases represent a major health problem with an increasing wor... more Nowadays obesity and its related diseases represent a major health problem with an increasing worldwide prevalence. Hyperplasia and hypertrophy of adipocytes lead to an excessive fat accumulation that is not efficiently prevented by current pharmacological treatments. So the research on anti-obesity drugs with good efficacy and tolerability able both to prevent and to reduce fat accumulation is of pivotal interest. In the present study we evaluated in vitro the effects of Valproic Acid, Berberin and Resveratrol on adipogenesis. Our experimental model was represented by human Mesenchymal Stem Cells (hMSCs), physiological precursors of adipocytes that can differentiate into adipocytes also in vitro. Preliminary cytotoxicity assays were performed in order to choose non-toxic doses of the three drugs. hMSCs were induced to adipogenic differentiation and treated with Valproic Acid, Berberin and Resveratrol at the selected doses. Controls were represented by hMSCs treated for adipogenesis...

Biomedical Science and Engineering, 2021

The innovative in silico technologies developed at SPILLOproject,1 e.g., the SPILLO potential bin... more The innovative in silico technologies developed at SPILLOproject,1 e.g., the SPILLO potential binding sites searcher (SPILLO-PBSS) software,2,3 allow to identify targets and off-targets of any small molecule on a multiple-organism proteomewide scale, and to perform an accurate multilevel cross-organism transferability analysis (MCOTA) aimed at rationalising animal testing. SPILLO-PBSS has been successfully used in several research projects, such as a study in which a compound (MV1035) was found to reduce migration and invasiveness in U87 glioblastoma (GBM) cell lines: the human structural proteome was analyzed and the RNA demethylase ALKBH5 has been identified as a target responsible for the observed effects (target experimentally validated). Another top-ranked target identified by SPILLO-PBSS, the DNA repair protein AlkB homolog 2 (ALKBH2), abundantly expressed in GBM cell lines, resulted particularly interesting for its pivotal role in the onset of resistance to Temozolomide (TMZ)...

Molecules, 2019

Multiple myeloma (MM) belongs to hematological cancers and its incidence is increasing worldwide.... more Multiple myeloma (MM) belongs to hematological cancers and its incidence is increasing worldwide. Despite recent advances in its therapy, MM still causes many deaths every year. In fact, current therapies sometimes fail and are associated with severe adverse effects, including neurotoxicity. As a part of our ongoing efforts to discover new potential therapies against MM, we prepared Hibiscus sabdariffa extracts obtained by a microwave-assisted solvent extraction and investigate their activity by in vitro assays on the RPMI-8226 cell line. The bioguided fractionation of the crude ethanolic extract allowed the identification of HsFC as the most effective extract. We assessed cell viability (MTT and Tripan blue test), cell migration (Boyden chamber assay), and neurotoxicity (DRG neurotoxicity assay). The promising results prompted us to further fractionate HsFC and we obtained two molecules effective against RPMI-8226 cells without neurotoxic effects at their active concentrations. Mor...

The FASEB Journal, 1999

Repeated intracerebroventricular (i.c.v.) microinjection of tumor necrosis factor-␣ (TNF-␣) into ... more Repeated intracerebroventricular (i.c.v.) microinjection of tumor necrosis factor-␣ (TNF-␣) into normal rats causes intramyelin and interstitial edema in the white matter of the spinal cord (SC). This response is identical to that observed in the SC white matter of rats made cobalamin (Cbl) deficient by total gastrectomy (TG). Immunoblot analysis showed that: 1) the level of the biologically active form of the TNF-␣ protein (17 kDa) is higher in the SC of totally gastrectomized (TGX) rats 2 months after TG, i.e., at the postoperative time when edema is observed; 2) SC levels of TNF-␣ protein (17 kDa) in 2-mo-TGX-, Cbl-treated rats are reduced to control. Repeated i.c.v. microinjections of anti-TNF-␣ antibodies, transforming growth factor-␤ 1 (TGF-␤ 1) or interleukin-6 (IL-6) into TGX rats, begun shortly after TG, substantially reduced both intramyelin and interstitial edema in the SC white matter. This study provides the first evidence that the hallmark myelin damage of Cbl-deficient central neuropathy, which is a pure myelinolytic disease, is not caused directly by the withdrawal of the vitamin itself, but reflects enhanced production of the biologically active form of TNF-␣ by SC cells. This study thus supports the view that TGF-␤ 1 and IL-6 may act as neuroprotective agents in Cbl deficiency central neuropathy.

Italian journal of anatomy and embryology, 2010

Aims In order to develop therapeutic strategies to control obesity and the related pathologies, i... more Aims In order to develop therapeutic strategies to control obesity and the related pathologies, it is very important to deepen the knowledge of adipocytes biology and the mechanisms that control adipogenesis. In the present work we have studied the molecular mechanisms at the basis of the adipocytes differentiation using the human Mesenchymal Stem Cells (hMSCs), undifferentiated stem cells present in the bone marrow that are the physiological precursors of adipocytes in the organism. In particular we have focused our attention on the MAPKinases ERK1 and ERK2, that are involved in many biological and cellular processes. Methods hMSCs, obtained from iliac crest bone marrow, were induced to adipogenic differentiation by treatment with Adipogenic Induction Medium for 10 days (determination phase), then replaced with Adipogenic Maintenance Medium until the end of treatment at day 28 (terminal differentiation phase). hMSC adipogenic differentiation was evaluated by morphological and molecular techniques. Control cells were represented by hMSCs cultured in absence of adipogenic supplements. ERK1 and ERK2 expression and phosphorylation were evaluated by immunoblotting experiments. The specific ERK inhibitor U0126 was added to the adipogenic medium at different times during the adipogenic differentiation protocol. Results hMSC treated with adipogenic differentiation protocol showed lipid droplets that increased in number and size during the whole differentiation period. In treated hMSC both ERK1 and ERK2 phosphorylation was reduced in comparison to control hMSCs, but time and intensity of these reductions were different for the two isoforms. A decrease of the total amount of ERK1 was also observed. The presence of U0126 during the whole differentiation period hampered the adipogenic differentiation of hMSCs, as very few hMSCs showed the appearance of lipid droplets that were reduced both in number and size. When U0126 was administered only during the determination phase the number of hMSCs recruited in the differentiation program was reduced, while when U0126 was administered only in the terminal differentiation phase, hMSCs did not acquired a mature adipocytic phenotype. Conclusion In this work we demonstrate that ERK1 and ERK2 are important for hMSC adipogenic differentiation. Our results suggest that ERK1 and ERK2 play a key role in determining how many cells enter into the adipogenic differentiation program and acquire the phenotipycal and molecular characteristics of mature adipocytes.

Molecules, 2017

The Editorial Board of the Medicinal Chemistry section of the journal Molecules publishes here it... more The Editorial Board of the Medicinal Chemistry section of the journal Molecules publishes here its first Editorial, which has been prepared by highlighting, in sub-editorials of about one hundred words, some selected recently published articles that may have a profound impact on drug discovery and therapy.[...]

Journal of Bacteriology, 1993

We have cloned and sequenced the genomic regions encompassing the rho genes of Neisseria gonorrho... more We have cloned and sequenced the genomic regions encompassing the rho genes of Neisseria gonorrhoeae and Salmonella typhimurium. Rho factor of S. typhimurium has only three amino acid differences with respect to the Escherichia coli homolog. Northern (RNA) blots and primer extension experiments were used to characterize the N. gonorrhoeae rho transcript and to identify the transcription initiation and termination elements of this cistron. The function of the Rho factor of N. gonorrhoeae was investigated by complementation assays of rho mutants of E. coli and S. typhimurium and by in vivo transcription assays in polar mutants of S. typhimurium.

The new microbiologica, 1994

Molecular genetics appears to be the most promising approach to understanding the biology and pat... more Molecular genetics appears to be the most promising approach to understanding the biology and pathology of Chlamydia. This report focuses on the cloning and the protein expression of a DNA fragment from Chlamydia trachomatis DK20 chromosome. Results of hybridization experiments suggest that this sequence is specifically present within chlamydial DNA. The coding capacity of this DNA fragment is supported by the expression of a 26,000 m.w. peptide, in an Escherichia coli maxicell system.

Giornale di batteriologia, virologia ed immunologia

In this study the MicroTrak (Syva Co.) system has been evaluated in comparison with the isolation... more In this study the MicroTrak (Syva Co.) system has been evaluated in comparison with the isolation of Chlamydia trachomatis onto McCoy cell monolayer, testing 120 endocervical samples taken from an unselected women population. The overall results show a higher incidence of positivity in women under 30 years of age; a sensitivity of the immunofluorescence of 76.3% and a specificity of 89.8%. The agreement between the two methods is significative when we test symptomatic patients.

Giornale di batteriologia, virologia ed immunologia

The Authors have evaluated, by DNA-probe hybridization, the presence of CMV in the cellular fract... more The Authors have evaluated, by DNA-probe hybridization, the presence of CMV in the cellular fractions of different body fluids from immunocompromised patients. This specific methodology appear to be rapid to perform, adaptable to different clinical samples and sensitive. Even in absence of a supporting evaluation of specific antibodies, direct detection of virus, by DNA-probe, seems to be the most efficient and rapid approach to confirm the diagnosis either of active phase or reactivating of CMV infection.

International Journal of Molecular Sciences

Rigosertib is a small molecule in preclinical development that, due to its characteristics as a d... more Rigosertib is a small molecule in preclinical development that, due to its characteristics as a dual PLK1 and PI3K inhibitor, is particularly effective in counteracting the advance of different types of tumors. In this work, we evaluated the efficacy of Rigosertib and the expression of p53 in five different human tumor cell lines in vitro, A549 (lung adenocarcinoma), MCF-7 and MDA-MB231 (breast cancer cells), RPMI 8226 (multiple myeloma), and U87-MG (glioblastoma). We demonstrated that in all cell lines, the effect was dose- and time-dependent, but A549 cells were the most sensible to the treatment while higher concentrations were required for the most resistant cell line U87-MG. Moreover, the highest and lowest p53 levels have been observed, respectively, in A459 and U87-MG cells. The alterations in the cell cycle and in cell-cycle-related proteins were observed in A549 at lower concentrations than U87-MG. In conclusion, with this article we have demonstrated that Rigosertib has di...

Italian journal of anatomy and embryology, 2015

Oxygen and glucose deprivation (OGD) due to ischemic events or trauma in the brain result in neur... more Oxygen and glucose deprivation (OGD) due to ischemic events or trauma in the brain result in neuronal loss. The therapeutic approaches available inadequate and often the outcome is unfavorable for the patient or at least unpredictable. Stem cells could be useful for the treatment of OGD injured-neurons. Mesenchymal Stem Cells (MSCs), isolated from bone marrow as well as from various tissues, have poor immunogenicity and neuroprotective properties being able to alleviate ischemic brain injuries in animal models. The Endothelial Progenitor Cells (EPCs) are present at low frequencies both in the bone marrow and in the peripheral blood. They are thought to play a role in the recovery of cerebrovasculature integrity after stroke. In the present study we evaluated the potential neuroprotective effect of human MSCs and human EPCs on rat embryonic cortical neurons injured by OGD. OGD was induced by incubating the cortical neurons in a hypoxia chamber in a 95% N2 + 5% CO2 atmosphere at 37°C ...

Italian journal of anatomy and embryology = Archivio italiano di anatomia ed embriologia, 2012

The peripheral nervous system is a frequent target of toxic agents. The accurate identification o... more The peripheral nervous system is a frequent target of toxic agents. The accurate identification of the sites of neurotoxic action through the morphological characterization of reliable in vivo models or in vitro systems can give fundamental clues when investigating the pathogenesis and interpreting the clinical features of drug-induced neuropathy. The morphological approach has been used to investigate almost all the anticancer drugs able to induce chemotherapy-induced peripheral neurotoxicity, i.e. platinum drugs, antitubulins and proteasome inhibitors. No models have ever been described for thalidomide. This review demonstrates that any pathogenetic study on chemotherapy-induced peripheral neurotoxicity must be based on solid morphological observations obtained in reliable animal and in vitro models. This is particularly true in this setting, since the availability of tissues of human origin is extremely limited. In fact, peripheral (generally sural) nerve biopsies are never requi...

Hibiscus Sabdariffa (HS) is a plant commonly used in folk medicine (1). In recent years HS has ga... more Hibiscus Sabdariffa (HS) is a plant commonly used in folk medicine (1). In recent years HS has gained great interest due to its important antioxidant, anti-inflammatory and antitumoral properties. In our work, we evaluated the in vitro anticancer effects of HS extract against two different human breast cancer cell lines: estrogen receptor (ER) positive MCF-7 cells and ER negative MDA-MB-231 cells. We tested both total extract (HSE) and one fraction obtained by ethyl acetate extraction (HSEC). MTT assay and Trypan Blue vital count showed a dose and time dependent reduction of the viability in both cell lines treated with different concentrations of HSE or HSEC compared to untreated control cells. A significantly marked reduction was observed in MCF-7 cells treated with HSEC. On the basis of our results we used the concentrations of 7.5mg/ml and 3.5mg/ml respectively for HSE and HSEC. In order to evaluate ER involvement in HS effect, we analyzed the cellular localization of the recept...

The quantum-mechanical effects typical for single atoms or molecules can be reproduced in microme... more The quantum-mechanical effects typical for single atoms or molecules can be reproduced in micrometer-scale electric devices. In these systems the essential component is a small Josephson junction (JJ) consisting of two superconductors separated by a thin insulator. The quantum phenomena can be controlled in real time by external signals and have a great potential for novel applications. However, their fragility on uncontrolled disturbance caused by typical nearby environments is a drawback for quantum information science, but a virtue for detector technology. Motivated by this we have theoretically studied transistor kind of devices based on single-charge tunneling through small JJs. A common factor of the research is the analysis of the interplay between the coherent Cooperpair (charge carriers in the superconducting state) tunneling and incoherent environmental processes. In the first work we calculate the current due to incoherent Cooper-pair tunneling through a voltage-biased small JJ in series with large JJs and compare the results with recent experiments. We are able to reproduce the main experimental features and interpret these as traces of energy levels and energy bands of the mesoscopic device. In the second work we analyze a similar circuit (asymmetric single-Cooper-pair transistor) but under the assumption that the Cooper-pair tunneling is mainly coherent. This predicts new resonant transport voltages in the circuit due to higher-order processes. However, no clear traces of most of them are seen in the experiments, and similar discrepancy is present also in the case of the symmetric circuit. We continue to study this problem by modeling the interplay between the coherent and incoherent processes more accurately using a density-matrix approach. By this we are able to demonstrate that in typical conditions most of these resonances are indeed washed out by strong decoherence caused by the environment. We also analyze the contribution of three typical weakly interacting dissipative environments: electromagnetic environment, spurious charge fluctuators in the nearby insulating materials, and quasiparticles. In the last work we model the dynamics of a currentbiased JJ perturbed by a smaller JJ using a similar density-matrix approach. We demonstrate that the small JJ can be used also as a detector of the energy-band dynamics in a current biased JJ. The method is also used for modeling the charge transport in the Bloch-oscillating transistor.

Italian journal of anatomy and embryology, 2011

Nowadays obesity and its related diseases represent a major health problem with an increasing wor... more Nowadays obesity and its related diseases represent a major health problem with an increasing worldwide prevalence. Hyperplasia and hypertrophy of adipocytes lead to an excessive fat accumulation that is not efficiently prevented by current pharmacological treatments. So the research on anti-obesity drugs with good efficacy and tolerability able both to prevent and to reduce fat accumulation is of pivotal interest. In the present study we evaluated in vitro the effects of Valproic Acid, Berberin and Resveratrol on adipogenesis. Our experimental model was represented by human Mesenchymal Stem Cells (hMSCs), physiological precursors of adipocytes that can differentiate into adipocytes also in vitro. Preliminary cytotoxicity assays were performed in order to choose non-toxic doses of the three drugs. hMSCs were induced to adipogenic differentiation and treated with Valproic Acid, Berberin and Resveratrol at the selected doses. Controls were represented by hMSCs treated for adipogenesis...

Biomedical Science and Engineering, 2021

The innovative in silico technologies developed at SPILLOproject,1 e.g., the SPILLO potential bin... more The innovative in silico technologies developed at SPILLOproject,1 e.g., the SPILLO potential binding sites searcher (SPILLO-PBSS) software,2,3 allow to identify targets and off-targets of any small molecule on a multiple-organism proteomewide scale, and to perform an accurate multilevel cross-organism transferability analysis (MCOTA) aimed at rationalising animal testing. SPILLO-PBSS has been successfully used in several research projects, such as a study in which a compound (MV1035) was found to reduce migration and invasiveness in U87 glioblastoma (GBM) cell lines: the human structural proteome was analyzed and the RNA demethylase ALKBH5 has been identified as a target responsible for the observed effects (target experimentally validated). Another top-ranked target identified by SPILLO-PBSS, the DNA repair protein AlkB homolog 2 (ALKBH2), abundantly expressed in GBM cell lines, resulted particularly interesting for its pivotal role in the onset of resistance to Temozolomide (TMZ)...

Molecules, 2019

Multiple myeloma (MM) belongs to hematological cancers and its incidence is increasing worldwide.... more Multiple myeloma (MM) belongs to hematological cancers and its incidence is increasing worldwide. Despite recent advances in its therapy, MM still causes many deaths every year. In fact, current therapies sometimes fail and are associated with severe adverse effects, including neurotoxicity. As a part of our ongoing efforts to discover new potential therapies against MM, we prepared Hibiscus sabdariffa extracts obtained by a microwave-assisted solvent extraction and investigate their activity by in vitro assays on the RPMI-8226 cell line. The bioguided fractionation of the crude ethanolic extract allowed the identification of HsFC as the most effective extract. We assessed cell viability (MTT and Tripan blue test), cell migration (Boyden chamber assay), and neurotoxicity (DRG neurotoxicity assay). The promising results prompted us to further fractionate HsFC and we obtained two molecules effective against RPMI-8226 cells without neurotoxic effects at their active concentrations. Mor...

The FASEB Journal, 1999

Repeated intracerebroventricular (i.c.v.) microinjection of tumor necrosis factor-␣ (TNF-␣) into ... more Repeated intracerebroventricular (i.c.v.) microinjection of tumor necrosis factor-␣ (TNF-␣) into normal rats causes intramyelin and interstitial edema in the white matter of the spinal cord (SC). This response is identical to that observed in the SC white matter of rats made cobalamin (Cbl) deficient by total gastrectomy (TG). Immunoblot analysis showed that: 1) the level of the biologically active form of the TNF-␣ protein (17 kDa) is higher in the SC of totally gastrectomized (TGX) rats 2 months after TG, i.e., at the postoperative time when edema is observed; 2) SC levels of TNF-␣ protein (17 kDa) in 2-mo-TGX-, Cbl-treated rats are reduced to control. Repeated i.c.v. microinjections of anti-TNF-␣ antibodies, transforming growth factor-␤ 1 (TGF-␤ 1) or interleukin-6 (IL-6) into TGX rats, begun shortly after TG, substantially reduced both intramyelin and interstitial edema in the SC white matter. This study provides the first evidence that the hallmark myelin damage of Cbl-deficient central neuropathy, which is a pure myelinolytic disease, is not caused directly by the withdrawal of the vitamin itself, but reflects enhanced production of the biologically active form of TNF-␣ by SC cells. This study thus supports the view that TGF-␤ 1 and IL-6 may act as neuroprotective agents in Cbl deficiency central neuropathy.

Italian journal of anatomy and embryology, 2010

Aims In order to develop therapeutic strategies to control obesity and the related pathologies, i... more Aims In order to develop therapeutic strategies to control obesity and the related pathologies, it is very important to deepen the knowledge of adipocytes biology and the mechanisms that control adipogenesis. In the present work we have studied the molecular mechanisms at the basis of the adipocytes differentiation using the human Mesenchymal Stem Cells (hMSCs), undifferentiated stem cells present in the bone marrow that are the physiological precursors of adipocytes in the organism. In particular we have focused our attention on the MAPKinases ERK1 and ERK2, that are involved in many biological and cellular processes. Methods hMSCs, obtained from iliac crest bone marrow, were induced to adipogenic differentiation by treatment with Adipogenic Induction Medium for 10 days (determination phase), then replaced with Adipogenic Maintenance Medium until the end of treatment at day 28 (terminal differentiation phase). hMSC adipogenic differentiation was evaluated by morphological and molecular techniques. Control cells were represented by hMSCs cultured in absence of adipogenic supplements. ERK1 and ERK2 expression and phosphorylation were evaluated by immunoblotting experiments. The specific ERK inhibitor U0126 was added to the adipogenic medium at different times during the adipogenic differentiation protocol. Results hMSC treated with adipogenic differentiation protocol showed lipid droplets that increased in number and size during the whole differentiation period. In treated hMSC both ERK1 and ERK2 phosphorylation was reduced in comparison to control hMSCs, but time and intensity of these reductions were different for the two isoforms. A decrease of the total amount of ERK1 was also observed. The presence of U0126 during the whole differentiation period hampered the adipogenic differentiation of hMSCs, as very few hMSCs showed the appearance of lipid droplets that were reduced both in number and size. When U0126 was administered only during the determination phase the number of hMSCs recruited in the differentiation program was reduced, while when U0126 was administered only in the terminal differentiation phase, hMSCs did not acquired a mature adipocytic phenotype. Conclusion In this work we demonstrate that ERK1 and ERK2 are important for hMSC adipogenic differentiation. Our results suggest that ERK1 and ERK2 play a key role in determining how many cells enter into the adipogenic differentiation program and acquire the phenotipycal and molecular characteristics of mature adipocytes.