The effect of the oncogenic beta-blocker DL-1-(2-nitro-3-methyl-phenoxy)-3-tert-butyl-amino-propa... more The effect of the oncogenic beta-blocker DL-1-(2-nitro-3-methyl-phenoxy)-3-tert-butyl-amino-propan-2-ol (DL-ZAMI 1305) on intercellular communication between cultured cells was studied. Intercellular communication of Chinese hamster V79 cells was measured by the dye-transfer method in which the spread of intracellularly microinjected fluorescent probe, Lucifer Yellow CH, through gap-junctions was used as an index of intercellular communication. When V79 cells are cultured with non-toxic doses (1-60 micrograms/ml) of DL-ZAMI 1305, a significant inhibition of dye-transfer is observed after 4 h. The inhibition is dose-related and greater than 90% inhibition is seen at the dose of 50 micrograms/ml. When DL-ZAMI 1305 is added at 0 and 24 h of experiment, its inhibitory effect is maintained for at least 48 h at high doses (50-60 micrograms/ml), whereas for lower doses of DL-ZAMI 1305, some recovery is seen after 24 h incubation. These results are suggestive of a possible tumor-promoting activity of DL-ZAMI 1305; in vivo studies on this carcinogen are in progress.
The contemporary discovery of extremely versatile engineered nucleic acid‐binding proteins has tr... more The contemporary discovery of extremely versatile engineered nucleic acid‐binding proteins has transformed a brave new world in the genome‐editing scientific area. Clustered regularly interspaced short palindromic repeats (CRISPR)‐mediated programmable nucleic acid‐binding proteins have brought about a revolution in diagnostic platforms. The groundbreaking finding that bacteria and archaea that harbored prokaryotes have transmitted adaptive immunity through CRISPR and CRISPR‐associated (Cas) proteins has driven revolutionary advances in molecular biology. Importantly, advances in gene editing focus how expanding visions in CRISPR‐Cas biology are revolutionizing the area of molecular diagnostics for identifying DNA and RNA in emerging microbiological pathogens, for single nucleotide polymorphism (SNP) identifications, and for cell‐free mutation. Recent advances, such as improvements in multiplexing and quantitative capabilities as well as instrument‐free detection of nucleic acids, w...
TPA induces a time-dependent, rapid and transient inhibition of the IAR 20 rat liver epithelial c... more TPA induces a time-dependent, rapid and transient inhibition of the IAR 20 rat liver epithelial cells gap-junctional intercellular communication (GJIC). Light microscopy and ultrastructural immunolocalization of connexin 43 (ex 43) reveals that during the cell uncoupling the junctional structures disappear and the connexin proteins dislocate in the perikarial cytoplasm. Immunoblot analysis shows that cytoplasmic ex 43 is apparently intact and present in all its isoforms. The blockage of protein synthesis after ex 43 “internalization” prevents the recovery of IAR 20 functional coupling. The results indicate that the TPA-induced inhibition of IAR 20 GJIC may result from the disassembly of the gap-junctional structures, followed by an “internalization” of the connexin proteins, rather than from a simple closure of the junctional channels.
Mercury (Hg), one of the most diffused and hazardous organ-specific environmental contaminants, e... more Mercury (Hg), one of the most diffused and hazardous organ-specific environmental contaminants, exists in a wide variety of physical and chemical states, each of which with unique characteristics of target organ specificity. Exposure to Hg vapour and to organic mercurials specifically affects the CNS, while the kidney is the target organ for inorganic Hg compounds. Despite the increasing number of studies, the molecular bases of the nephrotoxic potential of Hg has not, up to now, been clarified, even if there is evidence suggesting that the ability of the metal to interact with proteins (thiol groups) or to generate oxygen radicals may play a major role. Within this context, the aim of the present study was to investigate, in vitro, the mechanism(s) of the early nephrotoxic potential of mercury chloride (HgCl2), one of the most diffused and biologically active mercury (Hg2+) compounds. For this purpose, two kidney-derived in vitro systems (the MDCK and the LLC-PK1 cell lines) were t...
SUMMARY «In vitro study of the mechanisms of mercury chloride nephrotoxicity». Objectives: Mercur... more SUMMARY «In vitro study of the mechanisms of mercury chloride nephrotoxicity». Objectives: Mercury (Hg), one of the most diffused and hazardous organ-specific environmental contaminants, exists in a wide variety of physical and chemical states, each of which with unique characteristics of target organ specificity. Exposure to Hg vapour and to organic mercurials specifically affects the CNS, while the kidney is the target organ for inorganic Hg compounds. Despite the increasing number of studies, the molecular bases ...
Journal of Bone & Joint Surgery, British Volume, May 1, 2009
Aims: Cartilage lesions do not repair spontaneously. Despite the high number of surgical and tiss... more Aims: Cartilage lesions do not repair spontaneously. Despite the high number of surgical and tissue engineering approaches that have been applied to promote articular cartilage (AC) regeneration, none of them, at present, have shown durable satisfactory results. In the present study an innovative technology, the RCCS™ bioreactor (Synthecon, Inc.), was employed to develop a 3D culture model suitable for the long-term in vitro maintenance of AC explants. Based on the principle of relative microgravity, the RCCS™ bioreactor, in fact ...
Long-term spaceflight affects almost all physiological systems in humans and considerable amount ... more Long-term spaceflight affects almost all physiological systems in humans and considerable amount of data revealed its serious impact on skeletal homeostasis. While a microgravity environment has been proved to induce significant mineral loss and bone fragility (affecting, specifically, cancellous weight-bearing bones), its effect on articular cartilage (AC) is poorly known. AC is an avascular tissue, composed of relatively few mechanosensitive cells (chondrocytes), that synthesize a mechanically functional extracellular matrix (ECM), composed of collagen, proteoglycans and other proteins. In response to physical factors (e.g. pressure and deformation) chondrocytes regulate AC histomorphology and function, and may affect bone tissue homeostasis. It is then likely that the absence of gravitational load should alter chondrocytes\u2019 activity. In the present study we investigated, in vitro, the effect of long-term exposure to a simulated microgravity condition (vector-averaged gravity) on whole explants of cancellous bone (rat tibial proximal epiphyses) and AC tissue (newborn rabbit knee\u2019s joint). Tissue explants were kept in culture for up to 4 weeks by the use of the Rotary Cell Culture System (RCCSTM) bioreactor, the unique device, operating on the Earth\u2019s surface, capable of successfully simulate a microgravity environment. The analysis of the structural/mechanical properties of cancellous bone explants was performed by a numerical model based on the Cell Method, applied to the 3D reconstruction of micro-computed tomography scans of the bone samples. With regard to AC tissue, functional and structural properties were studied by comparative cell viability, histochemical and molecular analyses performed on either the cellular component or on their ECM. The results obtained demonstrate that, while our RCCSTM-based culture system is able to preserve native tissue architecture and cells\u2019 viability all during the experimental procedure, long-term exposure to a microgravity environment may effectively alter bone cells\u2019 and articular chondrocytes\u2019 physiology
The process of identifying and approving a new drug is a time-consuming and expensive procedure. ... more The process of identifying and approving a new drug is a time-consuming and expensive procedure. One of the biggest issues to overcome is the risk of hepatotoxicity, which is one of the main reasons for drug withdrawal from the market. While animal models are the gold standard in preclinical drug testing, the translation of results into therapeutic intervention is often ambiguous due to interspecies differences in hepatic metabolism. The discovery of human induced Pluripotent Stem Cells (hiPSCs) and their derivatives has opened new possibilities for drug testing. We used mesenchymal stem cells and hepatocytes both derived from hiPSC, together with endothelial cells, to miniaturize the process of generating hepatic organoids. These organoids were then cultivated in vitro using both static and dynamic cultures. Additionally, we tested spheroids composed solely by induced hepatocytes. By miniaturizing the system, we demonstrated the possibility of maintaining the organoids, but not the...
The effect of the oncogenic beta-blocker DL-1-(2-nitro-3-methyl-phenoxy)-3-tert-butyl-amino-propa... more The effect of the oncogenic beta-blocker DL-1-(2-nitro-3-methyl-phenoxy)-3-tert-butyl-amino-propan-2-ol (DL-ZAMI 1305) on intercellular communication between cultured cells was studied. Intercellular communication of Chinese hamster V79 cells was measured by the dye-transfer method in which the spread of intracellularly microinjected fluorescent probe, Lucifer Yellow CH, through gap-junctions was used as an index of intercellular communication. When V79 cells are cultured with non-toxic doses (1-60 micrograms/ml) of DL-ZAMI 1305, a significant inhibition of dye-transfer is observed after 4 h. The inhibition is dose-related and greater than 90% inhibition is seen at the dose of 50 micrograms/ml. When DL-ZAMI 1305 is added at 0 and 24 h of experiment, its inhibitory effect is maintained for at least 48 h at high doses (50-60 micrograms/ml), whereas for lower doses of DL-ZAMI 1305, some recovery is seen after 24 h incubation. These results are suggestive of a possible tumor-promoting activity of DL-ZAMI 1305; in vivo studies on this carcinogen are in progress.
The contemporary discovery of extremely versatile engineered nucleic acid‐binding proteins has tr... more The contemporary discovery of extremely versatile engineered nucleic acid‐binding proteins has transformed a brave new world in the genome‐editing scientific area. Clustered regularly interspaced short palindromic repeats (CRISPR)‐mediated programmable nucleic acid‐binding proteins have brought about a revolution in diagnostic platforms. The groundbreaking finding that bacteria and archaea that harbored prokaryotes have transmitted adaptive immunity through CRISPR and CRISPR‐associated (Cas) proteins has driven revolutionary advances in molecular biology. Importantly, advances in gene editing focus how expanding visions in CRISPR‐Cas biology are revolutionizing the area of molecular diagnostics for identifying DNA and RNA in emerging microbiological pathogens, for single nucleotide polymorphism (SNP) identifications, and for cell‐free mutation. Recent advances, such as improvements in multiplexing and quantitative capabilities as well as instrument‐free detection of nucleic acids, w...
TPA induces a time-dependent, rapid and transient inhibition of the IAR 20 rat liver epithelial c... more TPA induces a time-dependent, rapid and transient inhibition of the IAR 20 rat liver epithelial cells gap-junctional intercellular communication (GJIC). Light microscopy and ultrastructural immunolocalization of connexin 43 (ex 43) reveals that during the cell uncoupling the junctional structures disappear and the connexin proteins dislocate in the perikarial cytoplasm. Immunoblot analysis shows that cytoplasmic ex 43 is apparently intact and present in all its isoforms. The blockage of protein synthesis after ex 43 “internalization” prevents the recovery of IAR 20 functional coupling. The results indicate that the TPA-induced inhibition of IAR 20 GJIC may result from the disassembly of the gap-junctional structures, followed by an “internalization” of the connexin proteins, rather than from a simple closure of the junctional channels.
Mercury (Hg), one of the most diffused and hazardous organ-specific environmental contaminants, e... more Mercury (Hg), one of the most diffused and hazardous organ-specific environmental contaminants, exists in a wide variety of physical and chemical states, each of which with unique characteristics of target organ specificity. Exposure to Hg vapour and to organic mercurials specifically affects the CNS, while the kidney is the target organ for inorganic Hg compounds. Despite the increasing number of studies, the molecular bases of the nephrotoxic potential of Hg has not, up to now, been clarified, even if there is evidence suggesting that the ability of the metal to interact with proteins (thiol groups) or to generate oxygen radicals may play a major role. Within this context, the aim of the present study was to investigate, in vitro, the mechanism(s) of the early nephrotoxic potential of mercury chloride (HgCl2), one of the most diffused and biologically active mercury (Hg2+) compounds. For this purpose, two kidney-derived in vitro systems (the MDCK and the LLC-PK1 cell lines) were t...
SUMMARY «In vitro study of the mechanisms of mercury chloride nephrotoxicity». Objectives: Mercur... more SUMMARY «In vitro study of the mechanisms of mercury chloride nephrotoxicity». Objectives: Mercury (Hg), one of the most diffused and hazardous organ-specific environmental contaminants, exists in a wide variety of physical and chemical states, each of which with unique characteristics of target organ specificity. Exposure to Hg vapour and to organic mercurials specifically affects the CNS, while the kidney is the target organ for inorganic Hg compounds. Despite the increasing number of studies, the molecular bases ...
Journal of Bone & Joint Surgery, British Volume, May 1, 2009
Aims: Cartilage lesions do not repair spontaneously. Despite the high number of surgical and tiss... more Aims: Cartilage lesions do not repair spontaneously. Despite the high number of surgical and tissue engineering approaches that have been applied to promote articular cartilage (AC) regeneration, none of them, at present, have shown durable satisfactory results. In the present study an innovative technology, the RCCS™ bioreactor (Synthecon, Inc.), was employed to develop a 3D culture model suitable for the long-term in vitro maintenance of AC explants. Based on the principle of relative microgravity, the RCCS™ bioreactor, in fact ...
Long-term spaceflight affects almost all physiological systems in humans and considerable amount ... more Long-term spaceflight affects almost all physiological systems in humans and considerable amount of data revealed its serious impact on skeletal homeostasis. While a microgravity environment has been proved to induce significant mineral loss and bone fragility (affecting, specifically, cancellous weight-bearing bones), its effect on articular cartilage (AC) is poorly known. AC is an avascular tissue, composed of relatively few mechanosensitive cells (chondrocytes), that synthesize a mechanically functional extracellular matrix (ECM), composed of collagen, proteoglycans and other proteins. In response to physical factors (e.g. pressure and deformation) chondrocytes regulate AC histomorphology and function, and may affect bone tissue homeostasis. It is then likely that the absence of gravitational load should alter chondrocytes\u2019 activity. In the present study we investigated, in vitro, the effect of long-term exposure to a simulated microgravity condition (vector-averaged gravity) on whole explants of cancellous bone (rat tibial proximal epiphyses) and AC tissue (newborn rabbit knee\u2019s joint). Tissue explants were kept in culture for up to 4 weeks by the use of the Rotary Cell Culture System (RCCSTM) bioreactor, the unique device, operating on the Earth\u2019s surface, capable of successfully simulate a microgravity environment. The analysis of the structural/mechanical properties of cancellous bone explants was performed by a numerical model based on the Cell Method, applied to the 3D reconstruction of micro-computed tomography scans of the bone samples. With regard to AC tissue, functional and structural properties were studied by comparative cell viability, histochemical and molecular analyses performed on either the cellular component or on their ECM. The results obtained demonstrate that, while our RCCSTM-based culture system is able to preserve native tissue architecture and cells\u2019 viability all during the experimental procedure, long-term exposure to a microgravity environment may effectively alter bone cells\u2019 and articular chondrocytes\u2019 physiology
The process of identifying and approving a new drug is a time-consuming and expensive procedure. ... more The process of identifying and approving a new drug is a time-consuming and expensive procedure. One of the biggest issues to overcome is the risk of hepatotoxicity, which is one of the main reasons for drug withdrawal from the market. While animal models are the gold standard in preclinical drug testing, the translation of results into therapeutic intervention is often ambiguous due to interspecies differences in hepatic metabolism. The discovery of human induced Pluripotent Stem Cells (hiPSCs) and their derivatives has opened new possibilities for drug testing. We used mesenchymal stem cells and hepatocytes both derived from hiPSC, together with endothelial cells, to miniaturize the process of generating hepatic organoids. These organoids were then cultivated in vitro using both static and dynamic cultures. Additionally, we tested spheroids composed solely by induced hepatocytes. By miniaturizing the system, we demonstrated the possibility of maintaining the organoids, but not the...
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Papers by Giovanna Mazzoleni