Daniela Bratosin - Academia.edu (original) (raw)
Papers by Daniela Bratosin
Jurnal Medical Aradean …, 2010
Comptes Rendus Biologies, Oct 1, 1997
AcadGmie des sciences / Elsevler. Paris Medical sciences / Sciences medicales Molecular mechanism... more AcadGmie des sciences / Elsevler. Paris Medical sciences / Sciences medicales Molecular mechanisms of erythrophagocytosis. Characterization of the senescent erythrocytes that are phagocytized by macrophages Mkanismes mokculaires de l'kythrophagocytose. Caractkisa tion de la population d'kythrocytes senescen ts voube 2 la phagocytose par les macrophages
Transfusion medicine relies on the possibility of storing red blood cells (RBCs) for a prolonged ... more Transfusion medicine relies on the possibility of storing red blood cells (RBCs) for a prolonged period of time. A better understanding of the nature of changes in stored RBCs may provide new strategies to improve the balance of benefits and transfusion's risks. In the light of recent results on the mechanism of programmed cell death of RBCs (erythroptosis), where numerous cellular changes are common hallmarks of the apoptotic phenomenon, we have developed new flow cytometric criteria to evaluate the viability of stored erythrocytes. In order to evaluate the possible benefic effect of low-intensity laser irradiation for increasing the blood preservation period, we irradiated the whole blood from healthy donors in specially designed bags for research purposes by MacoPharma Company containing SAGM medium, and conserved it for 3 weeks. LED (Light emitting diode) (λ=465nm, P=60mW, P/S=100mW/cm 2) and laser diode (λ=660nm, P=30mW, P/S=50mW/cm 2) in continuous wave have been used as sources of irradiation. Doses were ranged between 0 and 2J/cm 3. After that period RBCs were analyzed by flow cytometry for morphological changes (FSC/SSC), apoptosis/necrosis analysis (FITC-annexin-V labeling/PI) and viability using an original Calcein-AM method on a FACScan cytometer using CellQuest Pro software for acquisition and analysis. Complementary, the RBCs integrity was analyzed by scanning electron microscopy (SEM). Morphological changes assessment of red blood cells by flow cytometry and scanning electron microscopy showed a good preservation of the discoidal shape for the irradiated cells compared with control.
Cytometry, Oct 15, 1997
AcadGmie des sciences / Elsevler. Paris Medical sciences / Sciences medicales Molecular mechanism... more AcadGmie des sciences / Elsevler. Paris Medical sciences / Sciences medicales Molecular mechanisms of erythrophagocytosis. Characterization of the senescent erythrocytes that are phagocytized by macrophages Mkanismes mokculaires de l'kythrophagocytose. Caractkisa tion de la population d'kythrocytes senescen ts voube 2 la phagocytose par les macrophages
Transfusion medicine relies on the possibility of storing red blood cells (RBCs) for a prolonged ... more Transfusion medicine relies on the possibility of storing red blood cells (RBCs) for a prolonged period of time. A better understanding of the nature of changes in stored RBCs may provide new strategies to improve the balance of benefits and transfusion’s risks. In the light of recent results on the mechanism of programmed cell death of RBCs (erythroptosis), where numerous cellular changes are common hallmarks of the apoptotic phenomenon, we have developed new flow cytometric criteria to evaluate the viability of stored erythrocytes. In order to evaluate the possible benefic effect of low-intensity laser irradiation for increasing the blood preservation period, we irradiated the whole blood from healthy donors in specially designed bags for research purposes by MacoPharma Company containing SAGM medium, and conserved it for 3 weeks. LED (Light emitting diode) (λ=465nm, P=60mW, P/S=100mW/cm
Among all organisms in aquatic ecosystems, microalgae are key targets in pollution cases, for two... more Among all organisms in aquatic ecosystems, microalgae are key targets in pollution cases, for two basic reasons: their eco-physiological similarities with terrestrial plants (the potential sensitivity of the same metabolic processes) and their role as primary producers. Any change in the proliferation of the primary producers could provoke a global alteration in the equilibrium of the aquatic ecosystems. These characteristics support the use of the freshwater microalgae in laboratory toxicological assays. Flow cytometric analyses of Chlorella fusca cells were performed in a FACScan analyzer or Cytomics FC 500, both equipped with an argon-ion excitation laser (488 nm), detectors of forward (FSS) and side (SSC) light scatter and fluorescence detectors. For each analyzed parameter, data were recorded in a logarithmic scale and results were expressed as mean values obtained from histograms in arbitrary units. Fluorescence of chlorophyll a (>645 nm) was used as a FCM gate to exclude n...
The use of nanoparticles in biological and medical applications has increased in the last years. ... more The use of nanoparticles in biological and medical applications has increased in the last years. Understanding their potential toxicity and effect on the body is crucial before clinical use. The most used cell culture systems to test their cytotoxicity are of nonhuman origin. For assessing cytotoxicity of nanoparticles we developed a new experimental cell system, the RBCs, and we have evaluated the toxic effects by flow cytometric analysis. Recently, we and other researchers have shown that PCD of human erythrocyte is related to an apoptosis mechanism operating in the absence of mitochondria and lacking the terminal effectors of nuclear apoptosis (erythroptosis also called eryptosis) and to determine human RBC viability, we described a new rapid and sensitive calcein-AM assay by flow cytometric determination. Consequently, in the present study, to evaluate cell-nanoparticles interactions, human RBCs were exposed to different concentrations of nanoparticles and analyzed by flow cytom...
Autologous Cell Implantation (ACI) is a currently practiced cell-based therapy to repair cartilag... more Autologous Cell Implantation (ACI) is a currently practiced cell-based therapy to repair cartilage defects. Several strategies have been explored to expand the number of chondrocytes ex vivo. However, these methods are unable to provide sufficient quantity of chondrocytes with unaltered phenotype. To maintain the original phenotype in monolayer culture and to expand cell proliferation, primary human chondrocytes isolated by enzymatic digestion were cultured in a DMEM medium supplemented with Ac-Gly-Gly-OH dipeptide The aim of our study was to investigate and compare by flow cytometry the viability and the cell proliferation of chondrocytes obtained by culture in medium containing the dipeptide with the cells cultured in a classical system. The results we obtained provide that proliferation and viability of chondrocytes cultured in presence of DMEM medium containing Ac-Gly-Gly-OH were higher and thus can be used in the culture of chondrocytes devoted to reconstructive clinical proced...
Osteoarthritis (OA) is the most common of all joint diseases, but the molecular basis of its onse... more Osteoarthritis (OA) is the most common of all joint diseases, but the molecular basis of its onset and progression is controversial. One characteristic of cartilage is that some chondrocytes reside in areas rich in ECM and it is well known that glycoproteins are abundant on the cell surface and in cartilage ECM. Our working hypothesize was that modification of glycoconjugates contribute to pathogenesis of chondrocyte and we have undertaken a comparative study of normal and OA chondrocytes analysis with fluorescently labeled lectins. Our results clearly indicate that the loss of sialic acid and exposure of the penultimate β-galactose residues of OA chondrocytes glycoconjugates cell surfaces renders cells more susceptible to the membrane damage, which may trigger apoptosis. This could further clarify the relevance of the glycophenotype for chondrocyte function or its significance, if any, as a marker for the development of new chondroprotective agents.
L'invention concerne un procede d'evaluation de la phagocytose in vitro des erythrocytes ... more L'invention concerne un procede d'evaluation de la phagocytose in vitro des erythrocytes par des macrophages en culture, qui comprend un marquage fluorescent des erythrocytes, leur mise en contact avec une culture de macrophages et la mesure de la fluorescence des macrophages resultant de la phagocytose par cytofluorimetrie en flux. En particulier, l'application du procede a des erythrocytes a permis d'etablir une correlation entre le degre de phagocytose et la senescence des erythrocytes et, ainsi, d'appliquer cette evaluation au controle de routine de la qualite des erythrocytes destines aux transfusions sanguines ainsi qu'au suivi de l'amelioration de leurs conditions de stockage.
In ecotoxicological assessments, bioassays (ecotoxicity tests or biotests) are one of the main to... more In ecotoxicological assessments, bioassays (ecotoxicity tests or biotests) are one of the main tools, defined as methods which use living cells, tissues, organism or communities to assess exposurerelated effects of chemicals. The increasing complexity of environmental degradation requires an increase in the capacity of scientific approach in monitoring and notification as early as possible risks. Our own objective concerns the detection of aquatic environment pollution in Romania and particularly in the Danube basin. For assessing aquatic environment pollution degree or for assessing cytotoxicity or ecotoxicity of pollutants (heavy metals, nanoparticles, pesticides, etc.) we developed news experimental bioassays based on the use of viability and apoptosis biomarkers of Chlorella cells by flow cytometry. Our proposed bioassays could be rapid and very sensitive tests for in laboratory aquatic risk assessment and biomonitoring.
Romanian Biotechnological Letters, 2020
Carrier red blood cells (RBCs) can be used for vascular delivery of encapsulated drugs, biologica... more Carrier red blood cells (RBCs) can be used for vascular delivery of encapsulated drugs, biologicals and nanoparticles of various compositions with multiple advantages including bioavailability, biocompatibility, large carrier volumes and vascular residence time. Iron-based magnetic nanoparticles (MNPs), with size less than 15 nm became superparamagnetic and they can be used for both, diagnostic (imagistic) and therapeutic purposes, including as controlled release drug delivery systems. For these reasons, the objective of this study was to employ human erythrocytes for the encapsulation of carbon shell iron-based core nanoparticles (obtained by continuous wave infrared laser pyrolysis of Fe(CO)5 by a transient opening of cell membrane pores using different hypo-osmotic dialysis. In order to evaluate the encapsulation of nanoparticles into red blood cells (RBCs), normal, unloaded and loaded red blood cells (RBCs) were examined by Scanning/Transmission Electron Microscopy and Energy-dispersive X-ray spectroscopy analysis (EDX). Here, it is reported that these composite iron-based nanoparticles that were synthesized and tested could lead to their use in conjunction with the red blood cells (RBCs) as a promising agent in biomedical applications.
ABSTRACT. The human activities have a negative impact to the environment, consisting in the water... more ABSTRACT. The human activities have a negative impact to the environment, consisting in the water contamination with toxic products, heavy metals or with xenobiotic substances. Manufactured nanomaterials (nanoparticles, nanotubes, nanosheets and nanowires) ...
ABSTRACT. Polycythemia vera (PV) is characterized by an absolute increase in the red blood cell m... more ABSTRACT. Polycythemia vera (PV) is characterized by an absolute increase in the red blood cell mass, but the mechanisms are not completely understood. In this study, we identified by flow cytometric analysis morphological forms that deviate from the classical ...
… Vasile Goldis Arad, …, 2009
During fertilization, sperm utilizes a specific serine-dependent protease, known as acrosin, an e... more During fertilization, sperm utilizes a specific serine-dependent protease, known as acrosin, an enzyme with trypsin-like activity. In the present paper, we describe two rapid and sensitive flow cytometric assays we have developed for the determination of the activation level of proacrosin into acrosin in sperm using two original fluorescent staining. In the first method, the proportion of sperm with active acrosin was determined using the Alexa Fluor® 488 trypsin inhibitor conjugate (SBTI) and the presence of bound inhibitor in sperm was determined by flow cytometric analysis. In the second method we used (CBZ-Ala-Arg)2-R110 trypsin, a specific proteinase substrate for trypsin activity determination. Our results demonstrate that valuable results concerning the sperm quality can be obtained using the two new flow cytometric techniques.
1 National Institute for Biological Science Research and Development, Bucharest, Romania 2 ”Vasil... more 1 National Institute for Biological Science Research and Development, Bucharest, Romania 2 ”Vasile Goldis” Western University of Arad, Faculty of Biology, Arad, Romania 3 CFR 2 Hospital, Bucharest, Romania ABSTRACT. Articular cartilage repair remains a challenge for regenerative medicine. This review summarizes our research findings that have been undertaken, with a particular emphasis on those techniques that have been introduced into our laboratory, via in vitro and preclinical studies.
Jurnal Medical Aradean …, 2010
Comptes Rendus Biologies, Oct 1, 1997
AcadGmie des sciences / Elsevler. Paris Medical sciences / Sciences medicales Molecular mechanism... more AcadGmie des sciences / Elsevler. Paris Medical sciences / Sciences medicales Molecular mechanisms of erythrophagocytosis. Characterization of the senescent erythrocytes that are phagocytized by macrophages Mkanismes mokculaires de l'kythrophagocytose. Caractkisa tion de la population d'kythrocytes senescen ts voube 2 la phagocytose par les macrophages
Transfusion medicine relies on the possibility of storing red blood cells (RBCs) for a prolonged ... more Transfusion medicine relies on the possibility of storing red blood cells (RBCs) for a prolonged period of time. A better understanding of the nature of changes in stored RBCs may provide new strategies to improve the balance of benefits and transfusion's risks. In the light of recent results on the mechanism of programmed cell death of RBCs (erythroptosis), where numerous cellular changes are common hallmarks of the apoptotic phenomenon, we have developed new flow cytometric criteria to evaluate the viability of stored erythrocytes. In order to evaluate the possible benefic effect of low-intensity laser irradiation for increasing the blood preservation period, we irradiated the whole blood from healthy donors in specially designed bags for research purposes by MacoPharma Company containing SAGM medium, and conserved it for 3 weeks. LED (Light emitting diode) (λ=465nm, P=60mW, P/S=100mW/cm 2) and laser diode (λ=660nm, P=30mW, P/S=50mW/cm 2) in continuous wave have been used as sources of irradiation. Doses were ranged between 0 and 2J/cm 3. After that period RBCs were analyzed by flow cytometry for morphological changes (FSC/SSC), apoptosis/necrosis analysis (FITC-annexin-V labeling/PI) and viability using an original Calcein-AM method on a FACScan cytometer using CellQuest Pro software for acquisition and analysis. Complementary, the RBCs integrity was analyzed by scanning electron microscopy (SEM). Morphological changes assessment of red blood cells by flow cytometry and scanning electron microscopy showed a good preservation of the discoidal shape for the irradiated cells compared with control.
Cytometry, Oct 15, 1997
AcadGmie des sciences / Elsevler. Paris Medical sciences / Sciences medicales Molecular mechanism... more AcadGmie des sciences / Elsevler. Paris Medical sciences / Sciences medicales Molecular mechanisms of erythrophagocytosis. Characterization of the senescent erythrocytes that are phagocytized by macrophages Mkanismes mokculaires de l'kythrophagocytose. Caractkisa tion de la population d'kythrocytes senescen ts voube 2 la phagocytose par les macrophages
Transfusion medicine relies on the possibility of storing red blood cells (RBCs) for a prolonged ... more Transfusion medicine relies on the possibility of storing red blood cells (RBCs) for a prolonged period of time. A better understanding of the nature of changes in stored RBCs may provide new strategies to improve the balance of benefits and transfusion’s risks. In the light of recent results on the mechanism of programmed cell death of RBCs (erythroptosis), where numerous cellular changes are common hallmarks of the apoptotic phenomenon, we have developed new flow cytometric criteria to evaluate the viability of stored erythrocytes. In order to evaluate the possible benefic effect of low-intensity laser irradiation for increasing the blood preservation period, we irradiated the whole blood from healthy donors in specially designed bags for research purposes by MacoPharma Company containing SAGM medium, and conserved it for 3 weeks. LED (Light emitting diode) (λ=465nm, P=60mW, P/S=100mW/cm
Among all organisms in aquatic ecosystems, microalgae are key targets in pollution cases, for two... more Among all organisms in aquatic ecosystems, microalgae are key targets in pollution cases, for two basic reasons: their eco-physiological similarities with terrestrial plants (the potential sensitivity of the same metabolic processes) and their role as primary producers. Any change in the proliferation of the primary producers could provoke a global alteration in the equilibrium of the aquatic ecosystems. These characteristics support the use of the freshwater microalgae in laboratory toxicological assays. Flow cytometric analyses of Chlorella fusca cells were performed in a FACScan analyzer or Cytomics FC 500, both equipped with an argon-ion excitation laser (488 nm), detectors of forward (FSS) and side (SSC) light scatter and fluorescence detectors. For each analyzed parameter, data were recorded in a logarithmic scale and results were expressed as mean values obtained from histograms in arbitrary units. Fluorescence of chlorophyll a (>645 nm) was used as a FCM gate to exclude n...
The use of nanoparticles in biological and medical applications has increased in the last years. ... more The use of nanoparticles in biological and medical applications has increased in the last years. Understanding their potential toxicity and effect on the body is crucial before clinical use. The most used cell culture systems to test their cytotoxicity are of nonhuman origin. For assessing cytotoxicity of nanoparticles we developed a new experimental cell system, the RBCs, and we have evaluated the toxic effects by flow cytometric analysis. Recently, we and other researchers have shown that PCD of human erythrocyte is related to an apoptosis mechanism operating in the absence of mitochondria and lacking the terminal effectors of nuclear apoptosis (erythroptosis also called eryptosis) and to determine human RBC viability, we described a new rapid and sensitive calcein-AM assay by flow cytometric determination. Consequently, in the present study, to evaluate cell-nanoparticles interactions, human RBCs were exposed to different concentrations of nanoparticles and analyzed by flow cytom...
Autologous Cell Implantation (ACI) is a currently practiced cell-based therapy to repair cartilag... more Autologous Cell Implantation (ACI) is a currently practiced cell-based therapy to repair cartilage defects. Several strategies have been explored to expand the number of chondrocytes ex vivo. However, these methods are unable to provide sufficient quantity of chondrocytes with unaltered phenotype. To maintain the original phenotype in monolayer culture and to expand cell proliferation, primary human chondrocytes isolated by enzymatic digestion were cultured in a DMEM medium supplemented with Ac-Gly-Gly-OH dipeptide The aim of our study was to investigate and compare by flow cytometry the viability and the cell proliferation of chondrocytes obtained by culture in medium containing the dipeptide with the cells cultured in a classical system. The results we obtained provide that proliferation and viability of chondrocytes cultured in presence of DMEM medium containing Ac-Gly-Gly-OH were higher and thus can be used in the culture of chondrocytes devoted to reconstructive clinical proced...
Osteoarthritis (OA) is the most common of all joint diseases, but the molecular basis of its onse... more Osteoarthritis (OA) is the most common of all joint diseases, but the molecular basis of its onset and progression is controversial. One characteristic of cartilage is that some chondrocytes reside in areas rich in ECM and it is well known that glycoproteins are abundant on the cell surface and in cartilage ECM. Our working hypothesize was that modification of glycoconjugates contribute to pathogenesis of chondrocyte and we have undertaken a comparative study of normal and OA chondrocytes analysis with fluorescently labeled lectins. Our results clearly indicate that the loss of sialic acid and exposure of the penultimate β-galactose residues of OA chondrocytes glycoconjugates cell surfaces renders cells more susceptible to the membrane damage, which may trigger apoptosis. This could further clarify the relevance of the glycophenotype for chondrocyte function or its significance, if any, as a marker for the development of new chondroprotective agents.
L'invention concerne un procede d'evaluation de la phagocytose in vitro des erythrocytes ... more L'invention concerne un procede d'evaluation de la phagocytose in vitro des erythrocytes par des macrophages en culture, qui comprend un marquage fluorescent des erythrocytes, leur mise en contact avec une culture de macrophages et la mesure de la fluorescence des macrophages resultant de la phagocytose par cytofluorimetrie en flux. En particulier, l'application du procede a des erythrocytes a permis d'etablir une correlation entre le degre de phagocytose et la senescence des erythrocytes et, ainsi, d'appliquer cette evaluation au controle de routine de la qualite des erythrocytes destines aux transfusions sanguines ainsi qu'au suivi de l'amelioration de leurs conditions de stockage.
In ecotoxicological assessments, bioassays (ecotoxicity tests or biotests) are one of the main to... more In ecotoxicological assessments, bioassays (ecotoxicity tests or biotests) are one of the main tools, defined as methods which use living cells, tissues, organism or communities to assess exposurerelated effects of chemicals. The increasing complexity of environmental degradation requires an increase in the capacity of scientific approach in monitoring and notification as early as possible risks. Our own objective concerns the detection of aquatic environment pollution in Romania and particularly in the Danube basin. For assessing aquatic environment pollution degree or for assessing cytotoxicity or ecotoxicity of pollutants (heavy metals, nanoparticles, pesticides, etc.) we developed news experimental bioassays based on the use of viability and apoptosis biomarkers of Chlorella cells by flow cytometry. Our proposed bioassays could be rapid and very sensitive tests for in laboratory aquatic risk assessment and biomonitoring.
Romanian Biotechnological Letters, 2020
Carrier red blood cells (RBCs) can be used for vascular delivery of encapsulated drugs, biologica... more Carrier red blood cells (RBCs) can be used for vascular delivery of encapsulated drugs, biologicals and nanoparticles of various compositions with multiple advantages including bioavailability, biocompatibility, large carrier volumes and vascular residence time. Iron-based magnetic nanoparticles (MNPs), with size less than 15 nm became superparamagnetic and they can be used for both, diagnostic (imagistic) and therapeutic purposes, including as controlled release drug delivery systems. For these reasons, the objective of this study was to employ human erythrocytes for the encapsulation of carbon shell iron-based core nanoparticles (obtained by continuous wave infrared laser pyrolysis of Fe(CO)5 by a transient opening of cell membrane pores using different hypo-osmotic dialysis. In order to evaluate the encapsulation of nanoparticles into red blood cells (RBCs), normal, unloaded and loaded red blood cells (RBCs) were examined by Scanning/Transmission Electron Microscopy and Energy-dispersive X-ray spectroscopy analysis (EDX). Here, it is reported that these composite iron-based nanoparticles that were synthesized and tested could lead to their use in conjunction with the red blood cells (RBCs) as a promising agent in biomedical applications.
ABSTRACT. The human activities have a negative impact to the environment, consisting in the water... more ABSTRACT. The human activities have a negative impact to the environment, consisting in the water contamination with toxic products, heavy metals or with xenobiotic substances. Manufactured nanomaterials (nanoparticles, nanotubes, nanosheets and nanowires) ...
ABSTRACT. Polycythemia vera (PV) is characterized by an absolute increase in the red blood cell m... more ABSTRACT. Polycythemia vera (PV) is characterized by an absolute increase in the red blood cell mass, but the mechanisms are not completely understood. In this study, we identified by flow cytometric analysis morphological forms that deviate from the classical ...
… Vasile Goldis Arad, …, 2009
During fertilization, sperm utilizes a specific serine-dependent protease, known as acrosin, an e... more During fertilization, sperm utilizes a specific serine-dependent protease, known as acrosin, an enzyme with trypsin-like activity. In the present paper, we describe two rapid and sensitive flow cytometric assays we have developed for the determination of the activation level of proacrosin into acrosin in sperm using two original fluorescent staining. In the first method, the proportion of sperm with active acrosin was determined using the Alexa Fluor® 488 trypsin inhibitor conjugate (SBTI) and the presence of bound inhibitor in sperm was determined by flow cytometric analysis. In the second method we used (CBZ-Ala-Arg)2-R110 trypsin, a specific proteinase substrate for trypsin activity determination. Our results demonstrate that valuable results concerning the sperm quality can be obtained using the two new flow cytometric techniques.
1 National Institute for Biological Science Research and Development, Bucharest, Romania 2 ”Vasil... more 1 National Institute for Biological Science Research and Development, Bucharest, Romania 2 ”Vasile Goldis” Western University of Arad, Faculty of Biology, Arad, Romania 3 CFR 2 Hospital, Bucharest, Romania ABSTRACT. Articular cartilage repair remains a challenge for regenerative medicine. This review summarizes our research findings that have been undertaken, with a particular emphasis on those techniques that have been introduced into our laboratory, via in vitro and preclinical studies.