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Papers by Evguenia Bekman
iPSCs for Modeling Central Nervous System Disorders
Annals of Medicine
Introduction: It is estimated that 55% to 75% of individuals who experience a stroke have persist... more Introduction: It is estimated that 55% to 75% of individuals who experience a stroke have persistent impairment of the affected upper limb (UL) [1,2]. It is needed to identify training strategies allied with interactive systems for retraining motor function of the UL. Virtual reality (VR), using either immersive or nonimmersive technology, seems to be one of those promising strategies. Virtual reality allows patients to have close-to-reality experiences, providing them varied, engaging, and realistic experiences [3]. For the physiotherapist, the use of the interactive technologies is a challenge which can improve treatment adherence, allow new environments adapted to patient needs, abilities and goals, as well as different task options [4]. The objective of this analysis was to systematically review the benefits and limitations of VR towards motor recovery of upper limb in post-stroke population. Materials and methods: Randomised controlled trials were researched in Pubmed and PEDro databases, between January 2009 and January 2019, using the following keywords: "Virtual reality", "video games", "upper limb" and "stroke". We included articles that used immersive and nonimmersive technology in upper limb recovery after stroke, and which compared VR with others modalities We excluded all articles in which the patient received home based intervention or community rehabilitation programs. All included clinical trials had level of evidence equal or superior to 6 score, assessed by PEDro scale. Results: Fifteen studies met the inclusion criteria. Only three studies considered immersive VR. The training of functional tasks appears to provide the greatest benefits in upper extremity function with improvements in joint range of motion, hand motor function, grip strength, and dexterity. Two studies indicated that long-term improvements persist at followup. None of the studies reported any significant adverse effects. Discussion and Conclusions: There is moderate to high evidence that supports the beneficial effects of VR on stroke patient upper limb motor recovery. However, more studies are needed to determine what kind of VR systems are the most appropriate, particularly which ones may contribute or affect cortical reorganisation. It is also needed to identify the most adequate frequency, duration and intensity for the sessions.
Frontiers in Molecular Neuroscience
Journal of Visualized Experiments
The FEBS Journal
Angelman syndrome (AS) is an incurable neurodevelopmental disease caused by loss of function of t... more Angelman syndrome (AS) is an incurable neurodevelopmental disease caused by loss of function of the maternally inherited UBE3A gene. AS is characterized by a defined set of symptoms, namely severe developmental delay, speech impairment, uncontrolled laughter, and ataxia. Current understanding of the pathophysiology of AS relies mostly on studies using the murine model of the disease, although alternative models based on patient-derived stem cells are now emerging. Here, we summarize the literature of the last decade concerning the three major brain areas that have been the subject of study in the context of AS: hippocampus, cortex, and the cerebellum. Our comprehensive analysis highlights the major phenotypes ascribed to the different brain areas. Moreover, we also discuss the major drawbacks of current models and point out future directions for research in the context of AS, which will hopefully lead us to an effective treatment of this condition in humans.
Frontiers in Cellular Neuroscience
Frontiers in Cellular Neuroscience
Annals of Medicine
Results: GRP released in the cell culture media when co-expressed with GGCX, VKOR and furin in th... more Results: GRP released in the cell culture media when co-expressed with GGCX, VKOR and furin in the presence of vitamin K, is processed at the pro-peptide and contain Gla residues. EVs released by the insect cells in this system were shown to be loaded with GRP protein and mRNA, and capable of reducing ECM calcium deposition of calcifying VSMCs and the production of TNFa in THP1 monocyte/macrophage cells stimulated with LPS. Discussion and conclusions: While the successful production of human cGRP constitutes a major achievement, this innovative methodology will open new opportunities for the production of other biological active VKDPs. Furthermore, EVs loaded with GRP were shown to have anti-mineralizing and anti-inflammatory properties, with promising therapeutic potentialities for calcification-related chronic inflammatory diseases.
Annals of Medicine
View Crossmark data future, we will be able to confirm or not that inhibition of APOBEC3 by Vif s... more View Crossmark data future, we will be able to confirm or not that inhibition of APOBEC3 by Vif sensitize to chemotherapy the breast cancer cells, and hopefully decrease recurrent relapses. We believe that the use of Vif to inhibit APOBEC3 is a promising therapeutic tool model with application in breast cancer disease.
Stem Cells International
Human morphogenesis is a complex process involving distinct microenvironmental and physical signa... more Human morphogenesis is a complex process involving distinct microenvironmental and physical signals that are manipulated in space and time to give rise to complex tissues and organs. Advances in pluripotent stem cell (PSC) technology have promoted the in vitro recreation of processes involved in human morphogenesis. The development of organoids from human PSCs represents one reliable source for modeling a large spectrum of human disorders, as well as a promising approach for drug screening and toxicological tests. Based on the “self-organization” capacity of stem cells, different PSC-derived organoids have been created; however, considerable differences between in vitro-generated PSC-derived organoids and their in vivo counterparts have been reported. Advances in the bioengineering field have allowed the manipulation of different components, including cellular and noncellular factors, to better mimic the in vivo microenvironment. In this review, we focus on different examples of bio...
Oncotarget, Jan 15, 2017
We propose a novel single-deoxynucleoside-based assay that is easy to perform and provides accura... more We propose a novel single-deoxynucleoside-based assay that is easy to perform and provides accurate values for the absolute length (in units of time) of each of the cell cycle stages (G1, S and G2/M). This flow-cytometric assay takes advantage of the excellent stoichiometric properties of azide-fluorochrome detection of DNA substituted with 5-ethynyl-2'-deoxyuridine (EdU). We show that by pulsing cells with EdU for incremental periods of time maximal EdU-coupled fluorescence is reached when pulsing times match the length of S phase. These pulsing times, allowing labelling for a full S phase of a fraction of cells in asynchronous populations, provide accurate values for the absolute length of S phase. We characterized additional, lower intensity signals that allowed quantification of the absolute durations of G1 and G2 phases.Importantly, using this novel assay data on the lengths of G1, S and G2/M phases are obtained in parallel. Therefore, these parameters can be estimated with...
Bba Gene Struct Express, 2001
Two genes encoding conglutin gamma have been isolated from a Lupinus albus genomic library and se... more Two genes encoding conglutin gamma have been isolated from a Lupinus albus genomic library and sequenced. The expression of conglutin gamma was studied by partial amino acid sequencing of the mature seed protein and by nucleotide sequencing of reverse transcriptase-polymerase chain reaction products from various tissues during the plant life cycle.
Stem Cells International
Human morphogenesis is a complex process involving distinct microenvironmental and physical signa... more Human morphogenesis is a complex process involving distinct microenvironmental and physical signals that are manipulated in space and time to give rise to complex tissues and organs. Advances in pluripotent stem cell (PSC) technology have promoted the in vitro recreation of processes involved in human morphogenesis. The development of organoids from human PSCs represents one reliable source for modeling a large spectrum of human disorders, as well as a promising approach for drug screening and toxicological tests. Based on the “self-organization” capacity of stem cells, different PSC-derived organoids have been created; however, considerable differences between in vitro-generated PSC-derived organoids and their in vivo counterparts have been reported. Advances in the bioengineering field have allowed the manipulation of different components, including cellular and noncellular factors, to better mimic the in vivo microenvironment. In this review, we focus on different examples of bio...
Ontogenez
The polymerase chain reaction with degenerate primers corresponding to the most conservative amin... more The polymerase chain reaction with degenerate primers corresponding to the most conservative amino acids 16-21 (ELEKEF) and 49-54 (WFQNRR) of the Antennapedia class homeodomain was used for the amplification of cDNA from regenerating planarians (asexual race of Dugesia tigrina). A total of six new Antennapedia-like homeobox sequences, designated Dutarh-1-Dutarh-6 (Dugesia tigrina asexual race homeobox gene), were obtained. Their comparison with other homeobox genes using a "Genebee" software (the EMBL Data Library) showed that all sequences except Dutarh-6 belong to the Antennapedia class. Dutarh-6 is closely related to a recently described novel homeobox gene subfamily that includes mouse mesodermal homeobox genes Mox-1 and Mox-2 and rat homeobox gene Gax.
Objective: It is known that all materials and devices used in the oral cavity can trigger adverse... more Objective: It is known that all materials and devices used in the oral cavity can trigger adverse effects. Dental cements, essential for the success of oral rehabilitation, may be responsible for the release of substances that can induce negative effects on cells, including their death. Dental cements are present in the transition between restoration and tooth and, although in small areas, remain in contact with the oral tissues of patients for long periods of time. The aim of this study is to compare the cytotoxic effect of three luting cements, namely resin cement, resin-modified glass ionomer cement and glass ionomer cement, on the cells from gingival fibroblast cell line. Method: We used 3T3 fibroblast cells exposed to different dilutions of medium previously incubated for 24 hours with the 3 different cements. The cements were prepared according to the manufacturer and the resin cement was prepared in 2 different ways: light cured and non-light cured. After an incubation-period...
Objectives: Dental cements, essential for the success of oral rehabilitation, may be responsible ... more Objectives: Dental cements, essential for the success of oral rehabilitation, may be responsible for releasing substances might compromise the viability of cells in the oral cavity. Our previous work compared the toxicity effect of Relyx Unicem 2 Automixâ (resin cement), Ketac Cem Plusâ (resin-modified glass ionomer cement) and Ketac Cem Easymixâ (glass ionomer cement) using MTT assay. The aim of this study is to confirm the results obtained by an independent cristal violet assay and to assess genotoxicity of these cements to fibroblasts using the micronuclei assay. Methods: We used 3T3 fibroblast cells exposed to different dilutions of medium previously incubated for 24 hours with the three different cements that were prepared according to the manufacturer’s instructions. The resin cement was prepared in two ways: light cured and non-light cured. After 24 hours cell viability and genotoxicity were measured by cristal violet and micronuclei assays, respectively. Results: Results wer...
iPSCs for Modeling Central Nervous System Disorders
Annals of Medicine
Introduction: It is estimated that 55% to 75% of individuals who experience a stroke have persist... more Introduction: It is estimated that 55% to 75% of individuals who experience a stroke have persistent impairment of the affected upper limb (UL) [1,2]. It is needed to identify training strategies allied with interactive systems for retraining motor function of the UL. Virtual reality (VR), using either immersive or nonimmersive technology, seems to be one of those promising strategies. Virtual reality allows patients to have close-to-reality experiences, providing them varied, engaging, and realistic experiences [3]. For the physiotherapist, the use of the interactive technologies is a challenge which can improve treatment adherence, allow new environments adapted to patient needs, abilities and goals, as well as different task options [4]. The objective of this analysis was to systematically review the benefits and limitations of VR towards motor recovery of upper limb in post-stroke population. Materials and methods: Randomised controlled trials were researched in Pubmed and PEDro databases, between January 2009 and January 2019, using the following keywords: "Virtual reality", "video games", "upper limb" and "stroke". We included articles that used immersive and nonimmersive technology in upper limb recovery after stroke, and which compared VR with others modalities We excluded all articles in which the patient received home based intervention or community rehabilitation programs. All included clinical trials had level of evidence equal or superior to 6 score, assessed by PEDro scale. Results: Fifteen studies met the inclusion criteria. Only three studies considered immersive VR. The training of functional tasks appears to provide the greatest benefits in upper extremity function with improvements in joint range of motion, hand motor function, grip strength, and dexterity. Two studies indicated that long-term improvements persist at followup. None of the studies reported any significant adverse effects. Discussion and Conclusions: There is moderate to high evidence that supports the beneficial effects of VR on stroke patient upper limb motor recovery. However, more studies are needed to determine what kind of VR systems are the most appropriate, particularly which ones may contribute or affect cortical reorganisation. It is also needed to identify the most adequate frequency, duration and intensity for the sessions.
Frontiers in Molecular Neuroscience
Journal of Visualized Experiments
The FEBS Journal
Angelman syndrome (AS) is an incurable neurodevelopmental disease caused by loss of function of t... more Angelman syndrome (AS) is an incurable neurodevelopmental disease caused by loss of function of the maternally inherited UBE3A gene. AS is characterized by a defined set of symptoms, namely severe developmental delay, speech impairment, uncontrolled laughter, and ataxia. Current understanding of the pathophysiology of AS relies mostly on studies using the murine model of the disease, although alternative models based on patient-derived stem cells are now emerging. Here, we summarize the literature of the last decade concerning the three major brain areas that have been the subject of study in the context of AS: hippocampus, cortex, and the cerebellum. Our comprehensive analysis highlights the major phenotypes ascribed to the different brain areas. Moreover, we also discuss the major drawbacks of current models and point out future directions for research in the context of AS, which will hopefully lead us to an effective treatment of this condition in humans.
Frontiers in Cellular Neuroscience
Frontiers in Cellular Neuroscience
Annals of Medicine
Results: GRP released in the cell culture media when co-expressed with GGCX, VKOR and furin in th... more Results: GRP released in the cell culture media when co-expressed with GGCX, VKOR and furin in the presence of vitamin K, is processed at the pro-peptide and contain Gla residues. EVs released by the insect cells in this system were shown to be loaded with GRP protein and mRNA, and capable of reducing ECM calcium deposition of calcifying VSMCs and the production of TNFa in THP1 monocyte/macrophage cells stimulated with LPS. Discussion and conclusions: While the successful production of human cGRP constitutes a major achievement, this innovative methodology will open new opportunities for the production of other biological active VKDPs. Furthermore, EVs loaded with GRP were shown to have anti-mineralizing and anti-inflammatory properties, with promising therapeutic potentialities for calcification-related chronic inflammatory diseases.
Annals of Medicine
View Crossmark data future, we will be able to confirm or not that inhibition of APOBEC3 by Vif s... more View Crossmark data future, we will be able to confirm or not that inhibition of APOBEC3 by Vif sensitize to chemotherapy the breast cancer cells, and hopefully decrease recurrent relapses. We believe that the use of Vif to inhibit APOBEC3 is a promising therapeutic tool model with application in breast cancer disease.
Stem Cells International
Human morphogenesis is a complex process involving distinct microenvironmental and physical signa... more Human morphogenesis is a complex process involving distinct microenvironmental and physical signals that are manipulated in space and time to give rise to complex tissues and organs. Advances in pluripotent stem cell (PSC) technology have promoted the in vitro recreation of processes involved in human morphogenesis. The development of organoids from human PSCs represents one reliable source for modeling a large spectrum of human disorders, as well as a promising approach for drug screening and toxicological tests. Based on the “self-organization” capacity of stem cells, different PSC-derived organoids have been created; however, considerable differences between in vitro-generated PSC-derived organoids and their in vivo counterparts have been reported. Advances in the bioengineering field have allowed the manipulation of different components, including cellular and noncellular factors, to better mimic the in vivo microenvironment. In this review, we focus on different examples of bio...
Oncotarget, Jan 15, 2017
We propose a novel single-deoxynucleoside-based assay that is easy to perform and provides accura... more We propose a novel single-deoxynucleoside-based assay that is easy to perform and provides accurate values for the absolute length (in units of time) of each of the cell cycle stages (G1, S and G2/M). This flow-cytometric assay takes advantage of the excellent stoichiometric properties of azide-fluorochrome detection of DNA substituted with 5-ethynyl-2'-deoxyuridine (EdU). We show that by pulsing cells with EdU for incremental periods of time maximal EdU-coupled fluorescence is reached when pulsing times match the length of S phase. These pulsing times, allowing labelling for a full S phase of a fraction of cells in asynchronous populations, provide accurate values for the absolute length of S phase. We characterized additional, lower intensity signals that allowed quantification of the absolute durations of G1 and G2 phases.Importantly, using this novel assay data on the lengths of G1, S and G2/M phases are obtained in parallel. Therefore, these parameters can be estimated with...
Bba Gene Struct Express, 2001
Two genes encoding conglutin gamma have been isolated from a Lupinus albus genomic library and se... more Two genes encoding conglutin gamma have been isolated from a Lupinus albus genomic library and sequenced. The expression of conglutin gamma was studied by partial amino acid sequencing of the mature seed protein and by nucleotide sequencing of reverse transcriptase-polymerase chain reaction products from various tissues during the plant life cycle.
Stem Cells International
Human morphogenesis is a complex process involving distinct microenvironmental and physical signa... more Human morphogenesis is a complex process involving distinct microenvironmental and physical signals that are manipulated in space and time to give rise to complex tissues and organs. Advances in pluripotent stem cell (PSC) technology have promoted the in vitro recreation of processes involved in human morphogenesis. The development of organoids from human PSCs represents one reliable source for modeling a large spectrum of human disorders, as well as a promising approach for drug screening and toxicological tests. Based on the “self-organization” capacity of stem cells, different PSC-derived organoids have been created; however, considerable differences between in vitro-generated PSC-derived organoids and their in vivo counterparts have been reported. Advances in the bioengineering field have allowed the manipulation of different components, including cellular and noncellular factors, to better mimic the in vivo microenvironment. In this review, we focus on different examples of bio...
Ontogenez
The polymerase chain reaction with degenerate primers corresponding to the most conservative amin... more The polymerase chain reaction with degenerate primers corresponding to the most conservative amino acids 16-21 (ELEKEF) and 49-54 (WFQNRR) of the Antennapedia class homeodomain was used for the amplification of cDNA from regenerating planarians (asexual race of Dugesia tigrina). A total of six new Antennapedia-like homeobox sequences, designated Dutarh-1-Dutarh-6 (Dugesia tigrina asexual race homeobox gene), were obtained. Their comparison with other homeobox genes using a "Genebee" software (the EMBL Data Library) showed that all sequences except Dutarh-6 belong to the Antennapedia class. Dutarh-6 is closely related to a recently described novel homeobox gene subfamily that includes mouse mesodermal homeobox genes Mox-1 and Mox-2 and rat homeobox gene Gax.
Objective: It is known that all materials and devices used in the oral cavity can trigger adverse... more Objective: It is known that all materials and devices used in the oral cavity can trigger adverse effects. Dental cements, essential for the success of oral rehabilitation, may be responsible for the release of substances that can induce negative effects on cells, including their death. Dental cements are present in the transition between restoration and tooth and, although in small areas, remain in contact with the oral tissues of patients for long periods of time. The aim of this study is to compare the cytotoxic effect of three luting cements, namely resin cement, resin-modified glass ionomer cement and glass ionomer cement, on the cells from gingival fibroblast cell line. Method: We used 3T3 fibroblast cells exposed to different dilutions of medium previously incubated for 24 hours with the 3 different cements. The cements were prepared according to the manufacturer and the resin cement was prepared in 2 different ways: light cured and non-light cured. After an incubation-period...
Objectives: Dental cements, essential for the success of oral rehabilitation, may be responsible ... more Objectives: Dental cements, essential for the success of oral rehabilitation, may be responsible for releasing substances might compromise the viability of cells in the oral cavity. Our previous work compared the toxicity effect of Relyx Unicem 2 Automixâ (resin cement), Ketac Cem Plusâ (resin-modified glass ionomer cement) and Ketac Cem Easymixâ (glass ionomer cement) using MTT assay. The aim of this study is to confirm the results obtained by an independent cristal violet assay and to assess genotoxicity of these cements to fibroblasts using the micronuclei assay. Methods: We used 3T3 fibroblast cells exposed to different dilutions of medium previously incubated for 24 hours with the three different cements that were prepared according to the manufacturer’s instructions. The resin cement was prepared in two ways: light cured and non-light cured. After 24 hours cell viability and genotoxicity were measured by cristal violet and micronuclei assays, respectively. Results: Results wer...