2,983 results on '"animal cell"'
Search Results
2. DNA damage induced during mitosis undergoes DNA repair synthesis
- Author
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Godinez, Veronica Gomez, Kabbara, Sami, Sherman, Adria, Wu, Tao, Cohen, Shirli, Kong, Xiangduo, Maravillas-Montero, Jose Luis, Shi, Zhixia, Preece, Daryl, Yokomori, Kyoko, and Berns, Michael W
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Biological Sciences ,Environmental Biotechnology ,Environmental Sciences ,Genetics ,Cancer ,Generic health relevance ,Animals ,Cell Line ,DNA ,DNA Breaks ,DNA Repair ,G1 Phase ,Humans ,Infrared Rays ,Lasers ,Mitosis ,Potoroidae ,ATM protein ,BRCA1 protein ,discoidin domain receptor ,DNA ligase ,DNA ligase IV ,gamma H2AX ,histone H2AX ,nibrin ,Rad51 protein ,tumor suppressor p53 binding protein 1 ,ubiquitin ,unclassified drug ,anaphase ,animal cell ,Article ,cell cycle G1 phase ,cell damage ,controlled study ,DNA damage ,DNA repair ,DNA synthesis ,double stranded DNA break ,homologous recombination ,human ,human cell ,metaphase ,mitosis ,nonhomologous end joining repair ,nonhuman ,potoroo ,Potorous tridactylus ,protein function ,protein localization ,regulatory mechanism ,adverse device effect ,adverse event ,animal ,biosynthesis ,cell line ,DNA strand breakage ,genetics ,infrared radiation ,laser ,radiation response ,rat kangaroo ,General Science & Technology - Abstract
Understanding the mitotic DNA damage response (DDR) is critical to our comprehension of cancer, premature aging and developmental disorders which are marked by DNA repair deficiencies. In this study we use a micro-focused laser to induce DNA damage in selected mitotic chromosomes to study the subsequent repair response. Our findings demonstrate that (1) mitotic cells are capable of DNA repair as evidenced by DNA synthesis at damage sites, (2) Repair is attenuated when DNA-PKcs and ATM are simultaneously compromised, (3) Laser damage may permit the observation of previously undetected DDR proteins when damage is elicited by other methods in mitosis, and (4) Twenty five percent of mitotic DNA-damaged cells undergo a subsequent mitosis. Together these findings suggest that mitotic DDR is more complex than previously thought and may involve factors from multiple repair pathways that are better understood in interphase.
- Published
- 2020
3. DNA damage induced during mitosis undergoes DNA repair synthesis.
- Author
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Gomez Godinez, Veronica, Kabbara, Sami, Sherman, Adria, Wu, Tao, Cohen, Shirli, Kong, Xiangduo, Maravillas-Montero, Jose Luis, Shi, Zhixia, Preece, Daryl, Yokomori, Kyoko, and Berns, Michael W
- Subjects
Cell Line ,Animals ,Potoroidae ,Humans ,DNA ,Lasers ,Infrared Rays ,Mitosis ,G1 Phase ,DNA Repair ,DNA Breaks ,General Science & Technology ,ATM protein ,BRCA1 protein ,discoidin domain receptor ,DNA ligase ,DNA ligase IV ,gamma H2AX ,histone H2AX ,nibrin ,Rad51 protein ,tumor suppressor p53 binding protein 1 ,ubiquitin ,unclassified drug ,anaphase ,animal cell ,Article ,cell cycle G1 phase ,cell damage ,controlled study ,DNA damage ,DNA repair ,DNA synthesis ,double stranded DNA break ,homologous recombination ,human ,human cell ,metaphase ,mitosis ,nonhomologous end joining repair ,nonhuman ,potoroo ,Potorous tridactylus ,protein function ,protein localization ,regulatory mechanism ,adverse device effect ,adverse event ,animal ,biosynthesis ,cell line ,DNA strand breakage ,genetics ,infrared radiation ,laser ,radiation response ,rat kangaroo - Abstract
Understanding the mitotic DNA damage response (DDR) is critical to our comprehension of cancer, premature aging and developmental disorders which are marked by DNA repair deficiencies. In this study we use a micro-focused laser to induce DNA damage in selected mitotic chromosomes to study the subsequent repair response. Our findings demonstrate that (1) mitotic cells are capable of DNA repair as evidenced by DNA synthesis at damage sites, (2) Repair is attenuated when DNA-PKcs and ATM are simultaneously compromised, (3) Laser damage may permit the observation of previously undetected DDR proteins when damage is elicited by other methods in mitosis, and (4) Twenty five percent of mitotic DNA-damaged cells undergo a subsequent mitosis. Together these findings suggest that mitotic DDR is more complex than previously thought and may involve factors from multiple repair pathways that are better understood in interphase.
- Published
- 2020
4. Introduction to Cell Biosensors Through 55 Years of Scientific Production
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Thouand, Gérald, Thouand, Gérald, editor, Belkin, Shimshon, Section Editor, Daunert, Sylvia, Section Editor, Freemont, Paul, Section Editor, Hermans, Julie, Section Editor, Karube, Isao, Section Editor, Martel, Sylvain, Section Editor, Michelini, Elisa, Section Editor, and Roda, Aldo, Section Editor
- Published
- 2022
- Full Text
- View/download PDF
5. Cell Manipulations by Optical Tweezers and Laser Ablation
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Hosokawa, Yoichiroh, Yasukuni, Ryohei, Yamada, Sohei, Sugiura, Tadao, Lu, Yongfeng, Section editor, and Sugioka, Koji, editor
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- 2021
- Full Text
- View/download PDF
6. Brain Endothelial Erythrophagocytosis and Hemoglobin Transmigration Across Brain Endothelium: Implications for Pathogenesis of Cerebral Microbleeds
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Chang, Rudy, Castillo, Juan, Zambon, Alexander C, Krasieva, Tatiana B, Fisher, Mark J, and Sumbria, Rachita K
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Biomedical and Clinical Sciences ,Neurosciences ,Brain Disorders ,Hematology ,Clinical Research ,Aetiology ,2.1 Biological and endogenous factors ,Neurological ,erythrophagocytosis ,cerebral microbleeds ,brain endothelial cells ,hemoglobin ,red blood cells ,transmigration ,Brain endothelial cells ,Cerebral microbleeds ,Erythrophagocytosis ,Hemoglobin ,Red blood cells ,Transmigration ,eosin ,hematoxylin ,lipocortin 5 ,phosphatidylserine ,sialidase ,adult ,animal cell ,animal experiment ,Article ,autofluorescence ,bEnd.3 cell line ,blood brain barrier ,brain blood vessel ,brain hemorrhage ,cell activation ,cell adhesion ,cell culture ,cell labeling ,cell migration ,confocal microscopy ,controlled study ,endothelium cell ,enzyme activity ,flow cytometry ,fluorescence microscopy ,image analysis ,immunofluorescence ,in vitro study ,internalization ,male ,microscopy ,mouse ,nonhuman ,nuclear magnetic resonance imaging ,oxidative stress ,pathogenesis ,transendothelial and transepithelial migration ,trypan blue assay ,Biochemistry and Cell Biology ,Biochemistry and cell biology ,Biological psychology - Abstract
Peripheral endothelial cells are capable of erythrophagocytosis, but data on brain endothelial erythrophagocytosis are limited. We studied the relationship between brain endothelial erythrophagocytosis and cerebral microhemorrhage, the pathological substrate of MRI-demonstrable cerebral microbleeds. To demonstrate the erythrophagocytic capability of the brain endothelium, we studied the interactions between brain endothelial cells and red blood cells exposed to oxidative stress in vitro, and developed a new in vitro cerebral microbleeds model to study the subsequent passage of hemoglobin across the brain endothelial monolayer. Using multiple approaches, our results show marked brain endothelial erythrophagocytosis of red blood cells exposed to oxidative stress compared with control red blood cells in vitro. This brain endothelial erythrophagocytosis was accompanied by passage of hemoglobin across the brain endothelial monolayer with unaltered monolayer integrity. In vivo and confocal fluorescence microscopy studies confirmed the extravasation of RBC exposed to oxidative stress across brain endothelium. These findings, demonstrating erythrophagocytosis mediated by the brain endothelial monolayer and the subsequent passage of iron-rich hemoglobin in vitro and RBC in vivo, may have implications for elucidating mechanisms involved in the development of cerebral microbleeds that are not dependent on disruption of the microvasculature.
- Published
- 2018
7. Techniques, challenges and future prospects for cell-based meat.
- Author
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Benny, Anmariya, Pandi, Kathiresan, and Upadhyay, Rituja
- Abstract
In response to a growing population and rising food demand, the food industry has come up with a wide array of alterations, innovations, and possibilities for making meat in vitro. In addition to revolutionizing the meat industry, this advancement also has profound effects on the environment, health, and welfare of animals. Thus, rather than using slaughtered animals, animal cells are employed to generate cell-based meat, with the cells' proliferation and differentiation taking place in the culture environment. The primary goal of this paper is to examine the overall mechanism and numerous approaches involved in the creation of cell-based meat. It also covers upcoming issues like technical, consumer, and regulatory issues, environmental concerns, the economy, cost of the product, health and safety concerns, and ethical, religious, and societal taboos. Finally, it assesses the future prospects of cell-based meat production. [ABSTRACT FROM AUTHOR]
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- 2022
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8. The cerium oxide nanoparticles toxicity induced physiological, histological and biochemical alterations in freshwater mussels, Unio crassus
- Author
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Arslan, P., Türkmen, E.U., Erkoç, F., Günal, A.Ç., Duran, H., Arslan, P., Türkmen, E.U., Erkoç, F., Günal, A.Ç., and Duran, H.
- Abstract
Introduction: Releasing of cerium oxide nanoparticles (nano-CeO2) to the nature has increased due to the widespread use in many fields ranging from cosmetics to the food industry. Therefore, nano-CeO2 has been included in the Organization for Economic Co-operation and Development's (OECD) priority list for engineering nanomaterials. In this study, the effects of nano-CeO2 on the freshwater mussels were investigated to reveal the impact on the freshwater systems on model organism. Methods: First, the chemical and structural properties of nano-CeO2 were characterized in details. Second, the freshwater mussels were exposed to environmentally relevant concentrations of nano-CeO2 as 10 mg, 25 mg and 50 mg/L during 48-h and 7-d. Third, after the exposure periods, hemolymph and tissue samples were taken to analyse the Total Hemocyte Counts (THCs) histology and oxidative stress parameters (total antioxidant status, glutathione, glutathione-S-transferase, and advanced oxidative protein products). Results: Significant decrease of the THCs was observed in the nano-CeO2 exposed mussels compared to the control group (P ;lt; 0.05). The histological results showed a positive association between nano-CeO2 exposure concentration in the water and level of tissue damage and histopathological alterations were detected in the gill and the digestive gland tissues. Oxidative stress parameters were slightly affected after exposure to nano-CeO2 (P ;gt; 0.05). In conclusion, this study showed that acute exposure of freshwater mussels to nano-CeO2 did not pose significant biological risk. However, it has been proven that mussels are able to accumulate nano-CeO2 significantly in their bodies. Conclusion: This suggests that nano-CeO2 may be a potential risk to other organisms in the ecosystem through trophic transfer in the food-web based on their habitat and niche in the ecosystem. © 2024 Elsevier GmbH
- Published
- 2024
9. Elastic ‘tethers’ connect separating anaphase chromosomes in a broad range of animal cells
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Forer, Arthur, Duquette, Michelle L, Paliulis, Leocadia V, Fegaras, E, Ono, M, Preece, D, and Berns, Michael W
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Genetics ,Rare Diseases ,Anaphase ,Animals ,Chromosome Segregation ,Diptera ,Kinetochores ,Male ,Mitosis ,Spermatocytes ,Spindle Apparatus ,Spindle structure ,Chromosome tethers ,Laser microbeam ,Chromosomes ,anaphase ,animal cell ,Article ,chromosome arm ,Ensifera ,mitosis ,nonhuman ,priority journal ,Pt K2 cell line ,spermatocyte ,spider ,animal ,chromosome segregation ,cytology ,genetics ,kinetochore ,male ,metabolism ,spindle apparatus ,Biochemistry and Cell Biology ,Plant Biology ,Developmental Biology ,Biochemistry and cell biology ,Plant biology - Abstract
We describe the general occurrence in animal cells of elastic components ("tethers") that connect individual chromosomes moving to opposite poles during anaphase. Tethers, originally described in crane-fly spermatocytes, exert force on chromosome arms opposite to the direction the anaphase chromosomes move. We show that they exist in a broad range of animal cells. Thus tethers are previously unrecognised components of general mitotic mechanisms that exert force on chromosomes and they need to be accounted for in general models of mitosis in terms of forces on chromosomes and in terms of what their roles might be.
- Published
- 2017
10. Rat embryonic hippocampus and induced pluripotent stem cell derived cultured neurons recover from laser-induced subaxotomy
- Author
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Selfridge, Aaron, Hyun, Nicholas, Chiang, Chai-Chun, Reyna, Sol M, Weissmiller, April M, Shi, Linda Z, Preece, Daryl, Mobley, William C, and Berns, Michael W
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Engineering ,Biomedical and Clinical Sciences ,Neurosciences ,Biomedical Engineering ,Stem Cell Research - Induced Pluripotent Stem Cell - Human ,Regenerative Medicine ,Stem Cell Research ,Stem Cell Research - Induced Pluripotent Stem Cell ,1.1 Normal biological development and functioning ,Neurological ,growth cone ,neurons ,induced pluripotent stem cell ,hippocampus ,subaxotomy ,regeneration ,Brain ,Cells ,Cytology ,Laser damage ,Mammals ,Neurodegenerative diseases ,Proteins ,Pulsed lasers ,Rats ,Repair ,Stem cells ,Growth cones ,Induced pluripotent stem cells ,Neurons ,actin ,tubulin ,animal cell ,animal experiment ,animal model ,Article ,axonal injury ,controlled study ,cytoskeletal remodeling ,cytoskeleton ,embryo ,human ,human cell ,immunofluorescence ,nerve cell culture ,nerve fiber regeneration ,nerve fiber transection ,neuronal growth cone ,nonhuman ,pluripotent stem cell ,quantitative analysis ,rat ,time series analysis ,Medical Biotechnology ,Biomedical engineering - Abstract
Axonal injury and stress have long been thought to play a pathogenic role in a variety of neurodegenerative diseases. However, a model for studying single-cell axonal injury in mammalian cells and the processes of repair has not been established. The purpose of this study was to examine the response of neuronal growth cones to laser-induced axonal damage in cultures of embryonic rat hippocampal neurons and induced pluripotent stem cell (iPSC) derived human neurons. A 532-nm pulsed [Formula: see text] picosecond laser was focused to a diffraction limited spot at a precise location on an axon using a laser energy/power that did not rupture the cell membrane (subaxotomy). Subsequent time series images were taken to follow axonal recovery and growth cone dynamics. After laser subaxotomy, axons thinned at the damage site and initiated a dynamic cytoskeletal remodeling process to restore axonal thickness. The growth cone was observed to play a role in the repair process in both hippocampal and iPSC-derived neurons. Immunofluorescence staining confirmed structural tubulin damage and revealed initial phases of actin-based cytoskeletal remodeling at the damage site. The results of this study indicate that there is a repeatable and cross-species repair response of axons and growth cones after laser-induced damage.
- Published
- 2015
11. In vitro quantification of melanoma tumor cell invasion.
- Author
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Hendrix, M J, Gehlsen, K R, Wagner, H N, Jr, Rodney, S R, Misiorowski, R L, and Meyskens, F L, Jr
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Amnion: ultrastructure ,Animals ,Basement Membrane: ultrastructure ,Cell Line ,Female ,Humans ,In Vitro Techniques ,Melanoma: pathology ,Methods ,Mice ,Mice ,Inbred C57BL ,Neoplasm Invasiveness: diagnosis ,Pregnancy ,Thymidine: metabolism ,radioisotope ,thymidine c 14 ,unclassified drug ,amnion ,animal cell ,article ,biological model ,cell culture ,cell invasion ,in vitro study ,melanoma ,mouse ,nonhuman ,pregnancy ,Amnion ,Animals ,Basement Membrane ,Cell Line ,Female ,Humans ,Melanoma ,Methods ,Mice ,Mice ,Inbred C57BL ,Neoplasm Invasiveness ,Pregnancy ,Thymidine - Abstract
In order to quantify the invasiveness of melanoma tumor cells in vitro, a modification of the amniotic basement membrane (BM) model, described by Liotta et al. (Cancer Letters, 11, 141, 1980), was used in combination with radiolabeled tumor cells. B16-F10 metastatic murine melanoma cells and a derived clone (B16-F10L) were prelabeled with 0.1 muCi/ml of [14C]thymidine for 20-24 h in serum-free medium at 37 degrees C. Following incubation, fetal bovine serum was added to a concentration of 5 per cent, and the cells were allowed to grow to confluency for the next 24-28 h. The labeled cells were seeded onto amniotic membranes situated in Membrane Invasion Culture System (MICS) chambers at a density of 2.5 X 10(4) per well. At various times points, radioactivity of tumor cells that completely traversed the membrane was determined using an under-the-membrane sampling method. The average percent invasion demonstrated by the B16-F10 line was 2.75 per cent, and 3.65 per cent exhibited by the B16-F10L cell line after 48-53 h in vitro. Since it was apparent that some variability in thickness existed among membrane samples, a morphological analysis was performed on five sectors of a three-inch-diameter sample from four different placentae. Differences and similarities in BM thickness within the same sector were noted by this technique and could possibly contribute to some variability observed in tumor cell invasion in this model. Another parameter examined was the proliferation of tumor cells in the upper and lower wells of the MICS chambers. By 48 h, approximately 32.1 per cent of the B16-F10 cell line as well as the clone had replicated in the upper wells associated with the BMs compared with a 32.9 per cent replication in the lower wells, which reaffirmed the viability of the tumor cells under experimental conditions and insured similarly replicating populations of cells. In order to quantify the invasiveness of radiolabeled tumor cells accurately through a biological membranous barrier, the proper concentration of cells must be used, tumor cell heterogeneity should be taken into consideration, the technique of sampling radiolabeled invasive cells should be critically analysed, and thickness of the membranous barrier should all be considered as possible important factors in the quantitative analyses.
- Published
- 2014
12. A Biophysical Analysis of Mitochondrial Movement: Differences Between Transport in Neuronal Cell Bodies Versus Processes
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Narayanareddy, Babu Reddy Janakaloti, Vartiainen, Suvi, Hariri, Neema, O'Dowd, Diane K, and Gross, Steven P
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Animals ,Lethal Dose 50 ,Poisoning: diagnosis ,Poisons: toxicity ,Rats ,Time Factors ,Axonal transport ,Mitochondrial motionfat droplet ,kinesin 1 ,molecular motor ,animal cell ,article ,biophysics ,brain ,central nervous system ,Drosophila ,intracellular transport ,microscopy ,mitochondrial permeability ,mitochondrion ,motion ,nerve cell ,nerve cell culture ,nerve fiber transport ,nonhuman ,priority journal ,velocity - Published
- 2014
13. Animal Cell Culture and Cryopreservation
- Author
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Gupta, Varsha, Sengupta, Manjistha, Prakash, Jaya, Tripathy, Baishnab Charan, Gupta, Varsha, Sengupta, Manjistha, Prakash, Jaya, and Tripathy, Baishnab Charan
- Published
- 2017
- Full Text
- View/download PDF
14. A dynamic model linking cell growth to intracellular metabolism and extracellular by‐product accumulation.
- Author
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Ramos, João R. C., Rath, Alexander G., Genzel, Yvonne, Sandig, Volker, and Reichl, Udo
- Abstract
Mathematical modeling of animal cell growth and metabolism is essential for the understanding and improvement of the production of biopharmaceuticals. Models can explain the dynamic behavior of cell growth and product formation, support the identification of the most relevant parameters for process design, and significantly reduce the number of experiments to be performed for process optimization. Few dynamic models have been established that describe both extracellular and intracellular dynamics of growth and metabolism of animal cells. In this study, a model was developed, which comprises a set of 33 ordinary differential equations to describe batch cultivations of suspension AGE1.HN.AAT cells considered for the production of α1‐antitrypsin. This model combines a segregated cell growth model with a structured model of intracellular metabolism. Overall, it considers the viable cell concentration, mean cell diameter, viable cell volume, concentration of extracellular substrates, and intracellular concentrations of key metabolites from the central carbon metabolism. Furthermore, the release of metabolic by‐products such as lactate and ammonium was estimated directly from the intracellular reactions. Based on the same set of parameters, this model simulates well the dynamics of four independent batch cultivations. Analysis of the simulated intracellular rates revealed at least two distinct cellular physiological states. The first physiological state was characterized by a high glycolytic rate and high lactate production. Whereas the second state was characterized by efficient adenosine triphosphate production, a low glycolytic rate, and reactions of the TCA cycle running in the reverse direction from α‐ketoglutarate to citrate. Finally, we show possible applications of the model for cell line engineering and media optimization with two case studies. [ABSTRACT FROM AUTHOR]
- Published
- 2020
- Full Text
- View/download PDF
15. Ultrastructural analysis of chemical synapses and gap junctions between Drosophila brain neurons in culture.
- Author
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Oh, Hyun-Woo, Campusano, Jorge M, Hilgenberg, Lutz G W, Sun, Xicui, Smith, Martin A, and O'Dowd, Diane K
- Subjects
Animals ,Animals ,Genetically Modified ,Animals ,Newborn ,Brain: cytology ,Cells ,Cultured ,Drosophila ,Drosophila Proteins: genetics ,metabolism ,Gap Junctions: metabolism ,ultrastructure ,Green Fluorescent Proteins: genetics ,metabolism ,Microscopy ,Electron: methods ,Neurons: ultrastructure ,Synapses: metabolism ,ultrasonography ,Cholinergic ,Electrical synapse ,Electron microscopy ,GABAergic ,Synapsin4 aminobutyric acid ,antibody ,protein bruchpilot ,synapsin ,unclassified drug ,animal cell ,animal tissue ,article ,brain electrophysiology ,brain nerve cell ,cell ultrastructure ,controlled study ,Drosophila ,electron microscopy ,GABAergic system ,gap junction ,immunohistochemistry ,interneuron ,nerve cell culture ,nerve regeneration ,neurite ,neurophysiology ,nonhuman ,presynaptic nerve ,priority journal ,synapse ,Animals ,Animals ,Genetically Modified ,Animals ,Newborn ,Brain ,Cells ,Cultured ,Drosophila ,Drosophila Proteins ,Gap Junctions ,Green Fluorescent Proteins ,Microscopy ,Electron ,Neurons ,Synapses - Abstract
Dissociated cultures from many species have been important tools for exploring factors that regulate structure and function of central neuronal synapses. We have previously shown that cells harvested from brains of late stage Drosophila pupae can regenerate their processes in vitro. Electrophysiological recordings demonstrate the formation of functional synaptic connections as early as 3 days in vitro (DIV), but no information about synapse structure is available. Here, we report that antibodies against pre-synaptic proteins Synapsin and Bruchpilot result in punctate staining of regenerating neurites. Puncta density increases as neuritic plexuses develop over the first 4 DIV. Electron microscopy reveals that closely apposed neurites can form chemical synapses with both pre- and postsynaptic specializations characteristic of many inter-neuronal synapses in the adult brain. Chemical synapses in culture are restricted to neuritic processes and some neurite pairs form reciprocal synapses. GABAergic synapses have a significantly higher percentage of clear core versus granular vesicles than non-GABA synapses. Gap junction profiles, some adjacent to chemical synapses, suggest that neurons in culture can form purely electrical as well as mixed synapses, as they do in the brain. However, unlike adult brain, gap junctions in culture form between neuronal somata as well as neurites, suggesting soma ensheathing glia, largely absent in culture, regulate gap junction location in vivo. Thus pupal brain cultures, which support formation of interneuronal synapses with structural features similar to synapses in adult brain, are a useful model system for identifying intrinsic and extrinsic regulators of central synapse structure as well as function.
- Published
- 2008
16. nAChR-mediated calcium responses and plasticity in Drosophila Kenyon cells.
- Author
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Campusano, Jorge M, Su, Hailing, Jiang, Shaojuan A, Sicaeros, Beatriz, and O'Dowd, Diane K
- Subjects
Animals ,Bungarotoxins: pharmacology ,Calcium: metabolism ,Calcium Channels: drug effects ,metabolism ,Calcium Signaling: drug effects ,physiology ,Cells ,Cultured ,Drosophila: metabolism ,Enzyme Inhibitors: pharmacology ,Female ,Male ,Mushroom Bodies: cytology ,drug effects ,metabolism ,Neural Pathways: drug effects ,metabolism ,Neuronal Plasticity: drug effects ,physiology ,Neurons: drug effects ,metabolism ,Nicotinic Agonists: pharmacology ,Receptors ,Nicotinic: drug effects ,metabolism ,Synaptic Transmission: drug effects ,physiology ,Thapsigargin: pharmacology ,Calcium imaging ,Kenyon cells ,Mushroom bodies ,nAChR ,Nicotinealpha bungarotoxin ,calcium ,cyclic AMP ,green fluorescent protein ,nicotine ,nicotinic receptor ,thapsigargin ,voltage gated calcium channel ,animal cell ,animal tissue ,article ,calcium cell level ,calcium transport ,cell population ,controlled study ,Drosophila ,evoked response ,female ,male ,mushroom body ,nerve cell plasticity ,nonhuman ,priority journal ,protein expression ,protein synthesis ,receptor upregulation ,signal transduction ,synaptic transmission ,Animals ,Bungarotoxins ,Calcium ,Calcium Channels ,Calcium Signaling ,Cells ,Cultured ,Drosophila ,Enzyme Inhibitors ,Female ,Male ,Mushroom Bodies ,Neural Pathways ,Neuronal Plasticity ,Neurons ,Nicotinic Agonists ,Receptors ,Nicotinic ,Synaptic Transmission ,Thapsigargin - Abstract
In Drosophila, nicotinic acetylcholine receptors (nAChRs) mediate fast excitatory synaptic transmission in mushroom body Kenyon cells, a neuronal population involved in generation of complex behaviors, including responses to drugs of abuse. To determine whether activation of nAChRs can induce cellular changes that contribute to functional plasticity in these neurons, we examined nicotine-evoked responses in cells cultured from brains of late stage OK107-GAL4 pupae. Kenyon cells can be identified by expression of green fluorescent protein (GFP+). Nicotine activates alpha-bungarotoxin-sensitive nAChRs, causing a rapid increase in intracellular calcium levels in over 95% of the Kenyon cells. The nicotine-evoked calcium increase has a voltage-gated calcium channel (VGCC) dependent component and a VGCC-independent component that involves calcium influx directly through nAChRs. Thapsigargin treatment reduces the nicotine response consistent with amplification by calcium release from intracellular stores. The response to nicotine is experience-dependent: a short conditioning pulse of nicotine causes a transient 50% reduction in the magnitude of the response to a test pulse of nicotine when the interpulse interval is 4 h. This cellular plasticity is dependent on activation of the VGCC-component of the nicotine response and on cAMP-signaling, but not on protein synthesis. These data demonstrate that activation of nAChRs induces a calcium-dependent plasticity in Kenyon cells that could contribute to adult behaviors involving information processing in the mushroom bodies including responses to nicotine.
- Published
- 2007
17. Stereological examination of the effect on the hippocampal dentate gyrus granule cells number of rats subjected to wireless internet during prenatal period: a preliminary study
- Author
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Murat Çetin RAGBETLİ, Büşranur ÖZALPER, Tahir CAKİR, and Rağbetli, Murat Çetin
- Subjects
brain weight ,xylene ,ketamine ,hippocampus ,Stereology ,animal experiment ,animal cell ,heparin ,Article ,General Biochemistry, Genetics and Molecular Biology ,animal tissue ,male ,Pregnancy ,dentate gyrus granular layer ,follow up ,controlled study ,rat ,Wi-Fi ,cell count ,Internet ,nonhuman ,hematoxylin ,General Medicine ,wireless communication ,Prenatal Rat ,Number of Granule Cells ,albino rat ,prenatal period ,preliminary data ,female ,eosin ,brain stem - Abstract
As a result of industrialisation and the many recent advances in technology, we are being intensely subjected to non-ionised radiation sources. As such, the effect of non-ionised radiation on human tissue is now being researched. In this study, the effects of Wi-Fi waves found in some third-generation mobile phones to facilitate internet connection on the brain hippocampal dentate gyrus granule cell quantity of rats during the prenatal period was stereologically examined. During the study, mated albino rats were subjected to Wi-Fi throughout pregnancy. A month after giving birth, six rats from each group, for a total of 12 rats, was sacrificed under perfusion and anaesthesia. Their skulls were opened and brains were removed to undergo routine checks in which the granule cells were counted. Strategic sections were defined, and every 45th section couple (with a thickness of 5 µm) was taken and dyed with haematoxylin and eosin (H&E) and cresyl violet stain. The granule cells were counted using a combination of the stereological disector method and Cavalieri’s principle. Then, the results were analysed using the Mann–Whitney U test. No statistically significant difference has been observed between the groups (p > 0.05). The findings were discussed in light of the relevant literature, and it was determined that throughout pregnancy, Wi-Fi modem device did not result in any changes in the dentate gyrus granule cell count of the hippocampus of postnatal rats’ brains. © 2022 Ondokuz Mayis Universitesi. All rights reserved. 2009-TFU-U075 The authors are grateful to Yuzuncu Yil University Department of Scientific Research Projects (2009-TFU-U075).
- Published
- 2022
18. Adoptive immunotherapy of BCR-ABL-induced chronic myeloid leukemia-like myeloproliferative disease in a murine model.
- Author
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Krause, Daniela S and Van Etten, Richard A
- Subjects
Animals ,Bone Marrow Transplantation ,Disease Models ,Animal ,Fusion Proteins ,bcr-abl ,Histocompatibility Testing ,Immunotherapy ,Adoptive ,Leukemia ,Myelogenous ,Chronic ,BCR-ABL Positive: immunology ,therapy ,Major Histocompatibility Complex ,Mice ,Mice ,Inbred BALB C ,Mice ,Inbred Strains ,Myeloproliferative Disorders: immunology ,therapy ,Transplantation Chimera ,BCR ABL protein ,allogenic bone marrow transplantation ,animal cell ,animal experiment ,animal model ,animal tissue ,article ,chimera ,chronic myeloid leukemia ,controlled study ,graft versus host reaction ,immunotherapy ,major histocompatibility complex ,male ,mouse ,myeloproliferative disorder ,nonhuman ,priority journal ,Retrovirus ,spleen cell ,Animals ,Bone Marrow Transplantation ,Disease Models ,Animal ,Fusion Proteins ,bcr-abl ,Histocompatibility Testing ,Immunotherapy ,Adoptive ,Leukemia ,Myeloid ,Chronic ,Major Histocompatibility Complex ,Mice ,Mice ,Inbred BALB C ,Mice ,Inbred Strains ,Myeloproliferative Disorders ,Transplantation Chimera - Abstract
Donor leukocyte infusion (DLI) can induce graft-versus-leukemia (GvL) reactions in patients with chronic myeloid leukemia (CML) relapsing after allogeneic bone marrow transplantation (BMT), but the mechanisms of the antileukemic effect of DLI are unknown, and the procedure is complicated by graft-versus-host disease (GvHD) and graft failure. Here, we adapted a murine retroviral BMT model of Philadelphia(+) leukemia by combining allogeneic bone marrow (BM) from C57Bl/6 (H-2(b)) mice with BCR-ABL-transduced Balb/c (H-2(d)) BM, inducing mixed chimerism and myeloproliferative disease in recipients resembling relapse of CML following allogeneic BMT. Infusions of allogeneic splenocytes eliminated BCR-ABL-induced CML-like disease in the majority of mixed chimeras, with significant GvL effects mediated by both CD4(+) and CD4(-) cells. BCR-ABL-induced acute B-lymphoblastic leukemia was also eradicated by DLI in major histocompatibility complex (MHC)-mismatched chimeras. Most DLI-treated mice converted to full allogeneic chimerism but succumbed frequently to GvHD or graft failure. When MHC-matched B10.D2 (H-2(d)) mice were the allogeneic donors, CML-like disease was more resistant to DLI. These results suggest that depletion of CD8(+) cells from DLI could impair GvL against CML, while increased MHC disparity between donor and recipient may improve the responsiveness of Philadelphia(+) B-lymphoblastic leukemia to DLI.
- Published
- 2004
19. Melanin as a target for melanoma chemotherapy: pro-oxidant effect of oxygen and metals on melanoma viability.
- Author
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Farmer, Patrick J, Gidanian, Shirley, Shahandeh, Babbak, Di Bilio, Angel J, Tohidian, Nilou, and Meyskens, Frank L, Jr
- Subjects
Cell Line ,Tumor ,Cell Survival ,Cells ,Cultured ,Copper: chemistry ,Cyclic N-Oxides: chemistry ,Dose-Response Relationship ,Drug ,Electrochemistry ,Electron Spin Resonance Spectroscopy ,Enzyme Inhibitors: chemistry ,Escherichia coli: metabolism ,Humans ,Hydrogen-Ion Concentration ,Indoles: chemistry ,Infant ,Newborn ,Male ,Melanins: metabolism ,Melanoma: drug therapy ,metabolism ,Metals: metabolism ,Oxidants: metabolism ,Oxidation-Reduction ,Oxidative Stress ,Oxygen: metabolism ,Plasmids: metabolism ,Reactive Oxygen Species ,Time Factors ,Zinc: chemistry ,Melanins ,Melanocytes ,Melanoma ,Metal uptake ,Oxygen ,Pro-oxidanthydroxyl radical ,melanin ,metal ,oxidizing agent ,animal cell ,cancer chemotherapy ,cell compartmentalization ,cell viability ,complex formation ,conference paper ,drug effect ,electron spin resonance ,gene targeting ,genetic susceptibility ,melanoma ,nonhuman ,oxidation reduction reaction ,oxidative stress ,oxygen consumption ,receptor affinity ,Cell Line ,Tumor ,Cell Survival ,Cells ,Cultured ,Copper ,Cyclic N-Oxides ,Dose-Response Relationship ,Drug ,Electrochemistry ,Electron Spin Resonance Spectroscopy ,Enzyme Inhibitors ,Escherichia coli ,Humans ,Hydrogen-Ion Concentration ,Indoles ,Infant ,Newborn ,Male ,Melanins ,Melanoma ,Metals ,Oxidants ,Oxidation-Reduction ,Oxidative Stress ,Oxygen ,Plasmids ,Reactive Oxygen Species ,Time Factors ,Zinc ,Animalia - Abstract
Melanoma cells have a poor ability to mediate oxidative stress, which may be attributed to constitutive abnormalities in their melanosomes. We hypothesize that disorganization of the melanosomes will allow chemical targeting of the melanin within. Chemical studies show that under oxidative conditions, synthetic melanins demonstrate increased metal affinity and a susceptibility to redox cycling with oxygen to form reactive oxygen species. The electron paramagnetic resonance (EPR)-active 5,5'-dimethyl-pyrollidine N-oxide spin adduct was used to show that binding of divalent Zn or Cu to melanin induces a pro-oxidant response under oxygen, generating superoxide and hydroxyl radicals. A similar pro-oxidant behaviour is seen in melanoma cell lines under external peroxide stress. Melanoma cultures grown under 95% O2/5% CO2 atmospheres show markedly reduced viability as compared with normal melanocytes. Cu- and Zn-dithiocarbamate complexes, which induce passive uptake of the metal ions into cells, show significant antimelanoma activity. The antimelanoma effect of metal- and oxygen-induced stress appears additive rather than synergistic; both treatments are shown to be significantly less toxic to melanocytes.
- Published
- 2003
20. Scenariusz lekcji zdalnej. Temat: Komórka roślinna i zwierzęca.
- Author
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WESOŁOWSKA-TURLEJ, ANNA
- Subjects
- *
YOUNG adults , *COVID-19 pandemic , *EDUCATIONAL change , *DIGITAL technology , *PLANT anatomy , *EDUCATIONAL technology , *ELECTRONIC textbooks - Abstract
In the spring of 2020, the COVID-19 pandemic resulted in the closure of schools all across the globe. In response to the spreading worldwide pandemic, education has changed dramatically, with the distinctive rise of e-learning, whereby teaching is undertaken remotely and on digital platforms. In Poland, this form of education is still in its infancy, even though its elements are being slowly introduced to the didactic process. In this article, the author presents a ready-to-use remote learning biology syllabus for fifth grade students using educational tools and platforms. The main intent is to promote and encourage the use of modern technologies as much as possible while working with students. The author proposes the following websites to familiarize young people with the structure of plant and animal cells: e-textbooks, learningapps.org, mindmeister.com and the Quiver 3 D Coloring App. [ABSTRACT FROM AUTHOR]
- Published
- 2019
21. Data-driven soft sensor for animal cell suspension culture process based on DRVM.
- Author
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Huang, Yonghong, Zang, Huan, Cheng, Xiaodong, Wu, Hongsheng, and Li, Jueyou
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CELL suspensions ,CELL culture ,LACTIC acid ,GLUCOSE analysis ,WEIGHT training ,MAXIMUM likelihood statistics ,DETECTORS - Abstract
Abstract In order to solve the problem that key state variables (such as glucose concentration, lactic acid concentration and cell density) in the dynamic process of animal cell suspension culture are difficult to be measured in real time, a data-driven soft sensor based on dynamic relevance vector machine (DRVM) algorithm is proposed. The dominant variables of the soft sensor model are selected according to the mechanisms process. The c o r r c o e f () function (belongs to the correlation coefficient command in MATLAB) is used to analyze the correlation among environmental variables, and the auxiliary variables of the soft sensor model are further determined. An improved method on the three edge location algorithm is used to optimize the dynamic weights of the DRVM model. Considering the influence of dynamic transition on soft sensor, the maximum likelihood distribution method under the Bayesian framework is used to train DRVM weight and super parameters, and the dynamic soft sensor model of animal cell suspension culture is established. The proposed method is applied to predict the key state variables in BHK-21 cell suspension culture Process. The experimental results show that compared with the traditional static soft sensing based on RVM, the data-driven soft sensor based on DRVM has higher accuracy, and the rationality and superiority of the method are verified. In order to further realize the real-time online prediction of key state variables, the monitoring interface of the suspension culture process is developed on the LabVIEW virtual instrument platform through its MATLAB Script node, and the data exchange of the DRVM soft sensor for the key state variables of the cell suspension culture process based on MATLAB and monitoring interface is realized. Highlights • A dynamic soft sensor model of animal cell suspension culture is proposed. • A novel location algorithm is designed to optimize the proposed model. • The proposed method is applied to real-time online prediction in BHK-21 suspension culture process. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
22. Glucocorticoids modulate the biosynthesis and processing of proThyrotropin releasing-hormone (proTRH)
- Author
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Bruhn, Thomas O, Huang, Susan S, Vaslet, Charles, and Nillni, Eduardo A
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Biomedical and Clinical Sciences ,Clinical Sciences ,Underpinning research ,1.1 Normal biological development and functioning ,Animals ,Cell Line ,Dexamethasone ,Gene Expression ,Glucocorticoids ,Peptide Fragments ,Pituitary Gland ,Anterior ,Protein Biosynthesis ,Protein Precursors ,Rats ,Thyrotropin-Releasing Hormone ,Transcription ,Genetic ,Transfection ,AtT20 cells ,glucocorticoids ,peptide biosynthesis ,proTRH ,TRH ,AtT 20 cells ,Peptide biosynthesis ,complementary dna ,dexamethasone ,glucocorticoid ,hormone receptor ,peptide fragment ,polyclonal antibody ,protirelin ,adenohypophysis ,amino terminal sequence ,animal cell ,article ,carboxy terminal sequence ,cell line ,gene expression ,genetic transfection ,hormonal regulation ,hormone synthesis ,nonhuman ,peptide synthesis ,priority journal ,protein degradation ,protein processing ,rna translation ,Paediatrics and Reproductive Medicine ,Endocrinology & Metabolism ,Clinical sciences - Abstract
The thyrotropin- (TRH) releasing hormone precursor (26 kDa) undergoes proteolytic cleavage at either of two sites, generating N-terminal 15 kDa/9.5 kDa or C-terminal 16.5/10 kDa intermediate forms that are processed further to yield five copies of TRH-Gly and seven non-TRH peptides. Glucocorticoids (Gcc) have been shown to enhance TRH gene expression in three different cell systems in vitro, an effect that occurs, at least in part, through transcriptional activation. Although this implies that an increase of TRH prohormone biosynthesis would take place, this had not been demonstrated as yet. We report here that the synthetic glucocorticoid dexamethasone (Dex) substantially elevated the de novo biosynthesis of the intact 26-kDa TRH prohormone and its intermediate products of processing in cultured anterior pituitary cells, an observation that is consistent with an overall upregulation of both the biosynthesis and degradation of the TRH precursor. We reasoned that Gcc may act not only at the transcriptional, but also at the translational/posttranslational level. To address this question we chose a different cell system, AtT20 cells transfected with a cDNA encoding preproTRH. Since TRH gene expression in these cells is driven by the CMV-IE promoter and not by an endogenous "physiological" promoter, these cells provide an ideal model to study selectively the effects of Gcc on the translation and posttranslational processing of proTRH without interference from a direct transcriptional activation of the TRH gene. Dex caused a significant 75.7% increase in newly synthesized 26-kDa TRH prohormone, suggesting that the glucocorticoid raised the translation rate. We then demonstrated that Dex treatment accelerated TRH precursor processing. Of interest, processing of the N- vs the C-terminal intermediate was influenced differentially by the glucocorticoid. Although the N-terminal intermediate product of processing accumulated, the C-terminal intermediate was degraded more rapidly. Consistent with these observations was the finding that the intracellular accumulation of the N-terminally derived peptide preproTRH 25-50 was enhanced, but levels of the C-terminally derived peptide preproTRH208-255 were reduced. Accumulation of TRH itself, whose five copies are N- and C-terminally derived, was also enhanced. We conclude that Gcc induce changes in the biosynthesis and processing of proTRH by increasing the translation rate and by differentially influencing the processing of N- vs C-terminal intermediates of the precursor molecule. These effects of Gcc at the translational and posttranslational levels result in an increase in TRH production accompanied by differential effects on the accumulation of N- and C-terminal non-TRH peptides.
- Published
- 1998
23. REPORT on the Fifth International Workshop on Chromosome 9 held at Eynsham, Oxfordshire, UK, September 4–6, 1996
- Author
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POVEY, S, ATTWOOD, J, CHADWICK, B, FREZAL, J, HAINES, JL, KNOWLES, M, KWIATKOWSKI, DJ, OLOPADE, OI, SLAUGENHAUPT, S, SPURR, NK, SMITH, M, STEEL, K, WHITE, JA, and PERICAK‐VANCE, MA
- Subjects
Genetics ,Animals ,Chromosome Aberrations ,Chromosome Disorders ,Chromosome Mapping ,Chromosomes ,Human ,Pair 9 ,Computational Biology ,Humans ,Mice ,Rats ,Species Specificity ,algorithm ,animal cell ,basal cell carcinoma ,bladder cancer ,cancer genetics ,cardiomyopathy ,chromosome 9 ,chromosome analysis ,chromosome map ,conference paper ,consensus sequence ,data analysis ,ehlers danlos syndrome ,friedreich ataxia ,gene location ,gene locus ,gene mapping ,hearing loss ,human ,human cell ,hypoplasia ,lymphatic leukemia ,marker gene ,meiosis ,mouse ,muscular dystrophy ,neuropathy ,nomenclature ,nonhuman ,priority journal ,sex determination ,tuberous sclerosis ,tumor suppressor gene ,workshop ,Clinical Sciences ,Genetics & Heredity - Abstract
The Fifth International workshop on chromosome 9 comprised a gathering of 36 scientists from seven countries and included a fairly even distribution of interests along chromosome 9 as well as a strong input from more global activities and from comparative mapping. At least eight groups had participated in the goal set at the previous workshop which was to improve the fine genetic mapping in different regions of chromosome 9 by meiotic breakpoint mapping in allocated regions and this has resulted in some greatly improved order information. Excellent computing facilities were available and all contributed maps were entered not only into SIGMA (and thence submitted to GDB) but also into a dedicated version of ACEDB which can be accessed on the Web in the form of one of 28 slices into which the chromosome has been arbitrarily divided. It was generally agreed that the amount of data is now overwhelming and that the integration and validation of all data is not only unrealistic in a short meeting but probably impossible until the whole chromosome has been sequenced and fully annotated. Sequence-ready contigs presented at the meeting totalled about 3 MB which is about one fiftieth of the estimated length. The single biggest barrier to integration of maps is the problem of non-standard nomenclature of loci. In the past 2 workshops efforts have been made to compare traditional 'consensus' maps made by human insight (still probably best for small specific regions) with those generated with some computer assistance (such as SIGMA) and those generated objectively by defined computer algorithms such as ldb. Since no single form of map or representation is entirely satisfactory for all purposes the maps reproduced in the published version of the report are confined to one of the genetic maps, in which Genethon and older markers have been incorporated, a Sigma map of the genes as symbols together with a listing of known 'disease' genes on chromosome 9, and a revised assessment of the mouse map together with a list of mouse loci predicted to be on human chromosome 9. One of the 25 ACEDB slices is also shown to illustrate strengths and weaknesses of this approach. Workshop files include not only all maps available at the time but also details of loci and details of the meiotic breakpoints in the CEPH families.
- Published
- 1997
24. Report on the 1996 International chromosome 9 workshop
- Author
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Povey, S, Attvvood, J, Chadwick, B, Frezal, J, Haines, JL, Knowles, M, Kwiatkowski, DJ, Olopade, OI, Slaugenhaupt, S, Spurr, NK, Smith, M, Steel, K, White, JA, and Pericak-Vance, MA
- Subjects
Genetics ,algorithm ,animal cell ,basal cell carcinoma ,bladder cancer ,cancer genetics ,cardiomyopathy ,chromosome 9 ,chromosome analysis ,chromosome map ,conference paper ,consensus sequence ,data analysis ,ehlers danlos syndrome ,friedreich ataxia ,gene location ,gene locus ,gene mapping ,hearing loss ,human ,human cell ,hypoplasia ,lymphatic leukemia ,marker gene ,meiosis ,mouse ,muscular dystrophy ,neuropathy ,nomenclature ,nonhuman ,priority journal ,sex determination ,tuberous sclerosis ,tumor suppressor gene ,workshop ,Genetics & Heredity ,Clinical Sciences - Abstract
The Fifth International workshop on chromosome 9 comprised a gathering of 36 scientists from seven countries and included a fairly even distribution of interests along chromosome 9 as well as a strong input from more global activities and from comparative mapping. At least eight groups had participated in the goal set at the previous workshop which was to improve the fine genetic mapping in different regions of chromosome 9 by meiotic breakpoint mapping in allocated regions and this has resulted in some greatly improved order information. Excellent computing facilities were available and all contributed maps were entered not only into SIGMA (and thence submitted to GDB) but also into a dedicated version of ACEDB which can be accessed on the Web in the form of one of 28 slices into which the chromosome has been arbitrarily divided. It was generally agreed that the amount of data is now overwhelming and that the integration and validation of all data is not only unrealistic in a short meeting but probably impossible until the whole chromosome has been sequenced and fully annotated. Sequence-ready contigs presented at the meeting totalled about 3 MB which is about one fiftieth of the estimated length. The single biggest barrier to integration of maps is the problem of non-standard nomenclature of loci. In the past 2 workshops efforts have been made to compare traditional 'consensus' maps made by human insight (still probably best for small specific regions) with those generated with some computer assistance (such as SIGMA) and those generated objectively by defined computer algorithms such as ldb. Since no single form of map or representation is entirely satisfactory for all purposes the maps reproduced in the published version of the report are confined to one of the genetic maps, in which Genethon and older markers have been incorporated, a Sigma map of the genes as symbols together with a listing of known 'disease' genes on chromosome 9, and a revised assessment of the mouse map together with a list of mouse loci predicted to be on human chromosome 9. One of the 25 ACEDB slices is also shown to illustrate strengths and weaknesses of this approach. Workshop files include not only all maps available at the time but also details of loci and details of the meiotic breakpoints in the CEPH families.
- Published
- 1997
25. Immunohistochemical distribution of Bcl-2 and p53 apoptotic markers in acetamiprid-induced nephrotoxicity
- Author
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Gokhan, Nur, Emrah, Caylak, Pinar Aksu, Kilicle, Safak, Sandayuk, Ozlem Onen, Celebi, Mühendislik ve Doğa Bilimleri Fakültesi -- Biyomedikal Mühendisliği Bölümü, and Nur, Gökhan
- Subjects
Protein bcl 2 ,Mouse ,Imidacloprid ,animal experiment ,Histopathology ,glomerulus ,animal cell ,Eosin ,Kidney ,Acetamiprid ,animal tissue ,Kidney distal tubule ,Neonicotinoids ,Immunoreactivity ,Toxic dose ,General & Internal Medicine ,Formaldehyde ,Bcl-2 ,Nicotinic Receptors ,Hematoxylin ,Nephrotoxicity ,Protein p53 ,Kidney hemorrhage ,P53 ,kidney capsule ,apoptosis ,Kidney tissue ,Kidney proximal tubule ,General Medicine ,Nonhuman ,Bioaccumulation ,immunohistochemistry ,Proximal tubule cell ,Degeneration ,Distal tubule cell ,Protein expression ,Kidney epithelium ,Atrophy ,Controlled study - Abstract
Pesticides, which adversely affect the critical metabolic processes of organisms, disrupt the physiological balance by specifically targeting enzymes and may lead to such consequences that may lead to death. It provides benefits in agricultural activities. The p53 protein antagonizes bcl-2, an anti-apoptotic protein character, and induces apoptosis by causing mitochondrial membrane permeability. This study aims to show the effect of acetamiprid, which is an insecticide from the neonicotinoid class, on bcl-2 and p53 immunoreactivity, which has an important place in the apoptotic mechanism in kidney tissue. A total of four groups including control and three experimental groups (the acetamiprid was administered 5, 10, and 15 mg kg−1) were formed in the study. After acetamiprid was administered via gavage for 14 days, the kidney tissues taken from the mice, which were sacrificed by cervical dislocation, were fixed in 10% formaldehyde solution for histological and immunohistochemical analyses, and as a result of routine tissue follow-up, the sections were blocked in paraffin and stained with haematoxylin–eosin and immunostaining. The histopathological examinations revealed that while the kidney tissue had a normal structure in the control group, degeneration in the distal and proximal tubules, glomerular degeneration, increase in the capsular area, glomerular atrophy, and haemorrhage were determined in the acetamiprid groups at increasing severity and frequency depending on the dose of the applied substance. In the kidney tissue, Bcl-2 and p53 immunoreactivity was observed in glomerular cells, sinusoidal epithelium, and proximal and distal tubule cells. The acetamiprid caused pathological changes in the kidneys in the dose range used. This effect also affects the expression of bcl-2 and p53 genes, which are biomarkers in the apoptotic mechanism. As acetamiprid accumulates in tissues, it increases the expression of p53 from cell death receptors, while suppressing the anti-apoptotic bcl-2 expression.
- Published
- 2022
26. Cell-specific regulation of agrin RNA splicing in the chick ciliary ganglion.
- Author
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Smith, M A and O'Dowd, D K
- Subjects
Agrin: genetics ,Alternative Splicing ,Animals ,Base Sequence ,Chick Embryo ,DNA ,Complementary ,Electrophysiology ,Ganglia ,Parasympathetic: embryology ,metabolism ,Gene Expression ,Molecular Sequence Data ,Polymerase Chain Reaction ,RNA ,Messenger: metabolism ,Receptors ,Cholinergic: chemistry ,agrin ,cholinergic receptor ,alternative rna splicing ,animal cell ,article ,cell activity ,cell aggregation ,cell specificity ,chicken ,ciliary ganglion ,gene expression ,genetic transcription ,nerve cell ,nonhuman ,priority journal ,rna splicing ,Agrin ,Alternative Splicing ,Animal ,Base Sequence ,Chick Embryo ,DNA ,Complementary ,Electrophysiology ,Ganglia ,Parasympathetic ,Gene Expression ,Molecular Sequence Data ,Polymerase Chain Reaction ,Receptors ,Cholinergic ,RNA ,Messenger ,Support ,U.S. Gov't ,P.H.S. - Abstract
Alternative splicing results in production of four agrin proteins (agrin0, agrin8, agrin11, and agrin19) with different AChR aggregating activities. However, the cellular origin of mRNAs encoding each agrin isoform remains unknown. Using single-cell PCR, we demonstrate that in the chick ciliary ganglion, nonneuronal cells express only mRNA encoding agrin0, whereas neurons express one or any combination of agrin mRNAs. Moreover, significant differences were observed between the agrin mRNA profiles of ciliary and choroid neurons in the ganglion. The abundance of each agrin mRNA, the fraction of neurons expressing each transcript, and the combinations of transcripts expressed by neurons also change during development. Our results demonstrate that transcripts encoding agrin proteins with high AChR aggregating activity are expressed exclusively by neurons in the ciliary ganglion and that alternative splicing of agrin mRNA is regulated during development and in a cell-specific manner.
- Published
- 1994
27. Potent Virucidal Activity In Vitro of Photodynamic Therapy with Hypericum Extract as Photosensitizer and White Light against Human Coronavirus HCoV-229E
- Author
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Beatriz Praena, Marta Mascaraque, Sabina Andreu, Raquel Bello-Morales, Edgar Abarca-Lachen, Valentina Rapozzi, Yolanda Gilaberte, Salvador González, José Antonio López-Guerrero, Ángeles Juarranz, Fundación de la Universidad Autónoma de Madrid, Instituto de Salud Carlos III, Praena, Beatriz, Mascaraque, Marta, Andreu, Sabina, Bello-Morales, Raquel, Gilaberte, Yolanda, González, Salvador, López-Guerrero, José Antonio, Juarranz, Ángeles, UAM. Departamento de Biología, and UAM. Departamento de Biología Molecular
- Subjects
Coronavirus ,Virucidal Activity ,coronavirus ,HCoV-229E ,Hypericum extract ,photosensitization ,virucide ,Huh-7 Cell Line ,Pharmaceutical Science ,White Light ,Virucide ,Photosensitization ,Biología y Biomedicina / Biología ,Animal Cell - Abstract
The emergent human coronavirus SARS-CoV-2 and its high infectivity rate has highlighted the strong need for new virucidal treatments. In this sense, the use of photodynamic therapy (PDT) with white light, to take advantage of the sunlight, is a potent strategy for decreasing the virulence and pathogenicity of the virus. Here, we report the virucidal effect of PDT based on Hypericum extract (HE) in combination with white light, which exhibits an inhibitory activity of the human coronavirus HCoV-229E on hepatocarcinoma Huh-7 cells. Moreover, despite continuous exposure to white light, HE has long durability, being able to maintain the prevention of viral infection. Given its potent in vitro virucidal capacity, we propose HE in combination with white light as a promising candidate to fight against SARS-CoV-2 as a virucidal compound., This research was funded by Fundación Universidad Autónoma de Madrid, grant number PI21/00315 and by Instituto de Salud Carlos iii, grant number PI21/00953.
- Published
- 2022
- Full Text
- View/download PDF
28. Functional complementation of ataxia-telangiectasia group D (AT-D) cells by microcell-mediated chromosome transfer and mapping of the AT-D locus to the region 11q22-23.
- Author
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Lambert, C, Schultz, RA, Smith, M, Wagner-McPherson, C, McDaniel, LD, Donlon, T, Stanbridge, EJ, and Friedberg, EC
- Subjects
Hybrid Cells ,Chromosomes ,Human ,Pair 11 ,Humans ,Ataxia Telangiectasia ,DNA Damage ,Chromosome Mapping ,Genetic Complementation Test ,Karyotyping ,Nucleic Acid Hybridization ,DNA Repair ,Phenotype ,In Vitro Techniques ,GENE TRANSFER ,HEREDITARY DISEASES ,IONIZING RADIATION ,CELL CYCLE ,DNA SYNTHESIS ,Chromosomes ,Human ,Pair 11 ,Cell cycle ,DNA synthesis ,Gene transfer ,Hereditary diseases ,Ionizing radiation ,animal cell ,article ,ataxia telangiectasia ,chromosome 11q ,chromosome 18 ,gene mapping ,gene transfer ,human ,human cell ,hybrid cell ,mouse ,nonhuman ,priority journal ,In Vitro ,Support ,Non-U.S. Gov't ,U.S. Gov't ,Non-P.H.S. ,P.H.S. ,Animalia ,Ataxia ,Ataxia telangiectasia ,Support ,Non-U.S. Gov't ,Support ,U.S. Gov't ,Non-P.H.S. ,Support ,U.S. Gov't ,P.H.S. - Abstract
The hereditary human disease ataxia-telangiectasia (AT) is characterized by phenotypic complexity at the cellular level. We show that multiple mutant phenotypes of immortalized AT cells from genetic complementation group D (AT-D) are corrected after the introduction of a single human chromosome from a human-mouse hybrid line by microcell-mediated chromosome transfer. This chromosome is cytogenetically abnormal. It consists primarily of human chromosome 18, but it carries translocated material from the region 11q22-23, where one or more AT genes have been previously mapped by linkage analysis. A cytogenetically normal human chromosome 18 does not complement AT-D cells after microcell-mediated transfer, whereas a normal human chromosome 11 does. We conclude that the AT-D gene is located on chromosome 11q22-23.
- Published
- 1991
29. Effects of fish oil on methotrexate-induced reproductive damage in rats
- Author
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Volkan Ipek, Kursat Kaya, Cigdem Cebi, Ali Gurel, and Leyla Elif Ozgu Ayozger
- Subjects
reproductive toxicity ,Male ,antioxidant ,sperm quality ,Urology ,thiobarbituric acid reactive substance ,animal experiment ,animal cell ,sperm count ,fish oil ,sperm viability ,sperm ,Thiobarbituric Acid Reactive Substances ,Article ,Antioxidants ,animal tissue ,histology ,Endocrinology ,Fish Oils ,biochemical analysis ,Semen ,caspase 3 ,Animals ,controlled study ,rat ,animal ,glutathione peroxidase ,semen parameters ,nonhuman ,Superoxide Dismutase ,animal model ,spermatozoon motility ,catalase ,apoptosis ,germ cell ,General Medicine ,general anesthesia ,Glutathione ,enzyme activity ,Rats ,Oxidative Stress ,Methotrexate ,immunohistochemistry ,histopathology ,Sperm Motility ,metabolism - Abstract
Methotrexate (MTX) is a folic acid antagonist that is commonly used in paediatric and adult oncology to treat a variety of malignancies. Internal organs, including the testis, are severely cytotoxic and genotoxic to MTX. Omega-3, as an antioxidant, has been shown to protect rat testis tissue from injury. The effect of fish oil (FO) on MTX-induced reproductive damage in rats was investigated in this work. The 28 animals were divided into four groups for this purpose (control, FO, MTX, and MTX-FO). On the third day, the MTX group received a single intraperitoneal injection of 20 mg/kg MTX. Furthermore, in the FO and MTX-FO groups, FO was delivered through gavage once daily for 14 days. All animals euthanized under general anaesthesia on the 15th day. TBARS, catalase, glutathione peroxidase, glutathione (GSH), superoxide dismutase levels were measured biochemically. The Cosentino grading system was utilized for histology. Germ cell thickness and caspase-3 activity were also evaluated. In addition, sperm motility rate, epididymal sperm count, aberrant sperm rate, and sperm vitality were measured to assess sperm quality. Some TBARS levels have increased, but GSH levels decreased significantly in the MTX group. FO reduced TBARS levels while considerably increasing GSH levels. All sperm quality measures were significantly lowered in the MTX group, while FO had a recovery effect. There were no notable variations in histopathology across groups except for germ cell thickness, which reduced considerably in the MTX group and recovered with FO treatment. As a result, FO has been shown to reduce testicular toxicity following MTX treatment in rats. © 2022 Wiley-VCH GmbH.
- Published
- 2022
30. Semi-Continuous Cultures as a Tool for Cell Line Characterization During Process Development
- Author
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Victores, S., Castillo, A., Faife, E., Rabasa, Y., Alvarez, Y., Rojas, L., Palacio, J., Figueredo, A., and Noll, Thomas, editor
- Published
- 2010
- Full Text
- View/download PDF
31. Comparison of Cell Culture Methods for Obtaining of rHU-EPO to Large Scale
- Author
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Ojito, E., Castro, A., Chea, M., Lugo, R., Suárez, E., Medina, A., Arias, M., Chico, E., and Noll, Thomas, editor
- Published
- 2010
- Full Text
- View/download PDF
32. Continuous cell flocculation for recombinant antibody harvesting.
- Author
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Burgstaller, Daniel, Krepper, Walpurga, Haas, Josselyn, Maszelin, Marine, Mohoric, Jure, Pajnic, Katja, Jungbauer, Alois, and Satzer, Peter
- Subjects
FLOCCULATION ,RECOMBINANT antibodies ,CONTINUOUS processing ,CELL separation ,AMMONIUM chloride - Abstract
Abstract: BACKGROUND: Integrated continuous production technology is of great interest in biopharmaceutical industry. Efficient, flexible and cost effective methods for continuous cell removal have to be developed, before a fully continuous and integrated product train can be realized. The paper describes the development and testing of such an integrated continuous and disposable set‐up for cell separation by flocculation combined with depth filtration. RESULTS: Screening of multiple flocculation agents, depth filters, and conditions demonstrated that the best performance was obtained with 0.0375% polydiallyldimethylammonium chloride (pDADMAC; a polycationic flocculation agent) in combination with Clarisolve® depth filters. Using this set‐up, a 4‐fold decrease of filtration area was achieved relative to standard filtration without flocculation, with yields of ≥97% and DNA depletion of up to 99%. Continuous operation was accomplished using a simple tubular reactor design with parallelization of the filtration. The reactor length was selected to allow a 13.2‐min residence time, which was sufficient to complete flocculation in batch experiments. Continuous flocculation performance was monitored on‐line using focused beam reflectance measurement. Filter switch cycles based on upstream pressure were controlled by in‐line pressure sensors, and were stable from one filter to the next. CONCLUSION: It was demonstrated that stable and efficient continuous flocculation associated with depth filtration can be easily accomplished using tubular reactors and parallelization. Continuous cell separation is essential for the development of fully continuous integrated process trains. This cost‐efficient disposable design run in continuous mode significantly reduces facility foot print, process costs and enables great flexbility. © 2017 The Authors. Journal of Chemical Technology & Biotechnology published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. [ABSTRACT FROM AUTHOR]
- Published
- 2018
- Full Text
- View/download PDF
33. The preventive role of Spirulina platensis (Arthrospira platensis) in immune and oxidative insults in a stress-induced rat model
- Author
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Nilay Seyidoglu, Cenk Aydin, Eda Köseli, and Rovshan Gurbanli
- Subjects
antioxidant ,Antioxidant ,medicine.medical_treatment ,Veterinary medicine ,antioxidant activity ,animal cell ,duodenum ,medicine.disease_cause ,Spirulina (Arthrospira) platensis ,0403 veterinary science ,chemistry.chemical_compound ,stress ,Corticosterone ,corticosterone blood level ,SF600-1100 ,oxidative stress ,oxidizing agent ,rat ,glutathione peroxidase ,physiological stress ,immune function ,Spirulina (genus) ,0303 health sciences ,Kidney ,biology ,Chemistry ,catalase ,malonaldehyde ,04 agricultural and veterinary sciences ,superoxide dismutase ,medicine.anatomical_structure ,oxidant–antioxidant status ,diet supplementation ,endoplasmic reticulum stress ,ileum ,gamma interferon ,stomach ,Research Article ,kidney ,medicine.medical_specialty ,oxidation ,040301 veterinary sciences ,brain ,animal experiment ,Arthrospira platensis ,blood vessel reactivity ,testis tissue ,interleukin 6 ,Ileum ,Immune function ,heart ,Oxidative phosphorylation ,Stress ,interleukin 2 ,immunization ,liver ,aryldialkylphosphatase ,interleukin 4 ,Article ,animal tissue ,body weight ,03 medical and health sciences ,Immune system ,male ,nitric oxide ,Internal medicine ,medicine ,spectrophotometry ,controlled study ,defense mechanism ,aryldialkylphosphatase 1 ,dianisidine ,030304 developmental biology ,spirulina (arthrospira) platensis ,Oxidant-antioxidant status ,nonhuman ,colon ,General Veterinary ,animal model ,cost effectiveness analysis ,corticosterone ,antioxidant assay ,biology.organism_classification ,microalga ,enzyme linked immunosorbent assay ,Endocrinology ,organ weight ,testosterone ,colorimetry ,spleen ,diet ,Oxidative stress - Abstract
Introduction There is a balance between oxidative stress, antioxidant capacity and immune response. Their roles in physiological and behavioural mechanisms are important for the maintenance of the organism’s internal equilibrium. This study aimed to evaluate the antioxidant effects of the exogenous alga Spirulina platensis (Arthrospira platensis) in a stress-induced rat model, and to describe its possible mechanism of action. Material and Methods Thirty-six adult male Sprague Dawley rats were separated into four groups: control (C), stress (S), S. platensis (Sp), and S. platensis + stress (SpS). The rats in groups Sp and SpS were fed with 1,500 mg/kg b.w./day Spirulina platensis for 28 days. All rats were exposed to prolonged light phase conditions (18 h light : 6 h dark) for 14 days. The SpS and S groups were exposed to stress by being kept isolated and in a crowded environment. Blood samples were obtained by puncturing the heart on the 28th day. The effect of stress on serum corticosterone, oxidative stress markers (TOS, TAC, PON1, OSI) and immunological parameters (IL-2, IL-4, IFN-ɣ) were tested. Also, the brain, heart, intestines (duodenum, ileum, and colon), kidney, liver, spleen, and stomach of the rats were weighed. Results Serum corticosterone levels were higher in the S group than in the C group, and significantly lower in the SpS group than in the S group. Mean total antioxidant capacity were lower in the S group than in the C group, and Spirulina reversed this change. Although not significantly different, IL-2 was lower in the S group than in the C group. However, in the SpS group, IL-2 increased due to Spirulina platensis mitigating effects of stress. Conclusion Male rats fed a diet with Spirulina platensis could experience significantly milder physiological changes during stress, although stress patterns may be different. Exogenous antioxidant supplements merit further investigation in animals and humans where the endogenous defence mechanism against stress may not be sufficient.
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- 2021
34. Antibody capture process based on magnetic beads from very high cell density suspension
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Kristofer Eriksson, Veronique Chotteau, Hubert Schwarz, Nils Arnold Brechmann, Per-Olov Eriksson, and Atefeh Shokri
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host cell ,Medical Biotechnology ,Cell ,animal cell ,Applied Microbiology and Biotechnology ,Biopharmaceuticals ,Adsorption efficiency ,Medicinsk bioteknologi ,magnetic separation ,Clarifiers ,Suspension (vehicle) ,Chemistry ,Chinese hamster ovary cell ,antibody isolation ,Bioprocessteknik ,Magnetism ,fed batch culture ,Biochemistry and Molecular Biology ,Antibodies, Monoclonal ,mAb capture ,High cell ,Monoclonal antibodies (mAbs) ,medicine.anatomical_structure ,Batch Cell Culture Techniques ,Process intensification ,Scientific method ,high cell density ,Antibody capture ,Biotechnology ,medicine.drug_class ,Cells ,Suspensions (components) ,Magnetic separation ,Bioengineering ,CHO Cells ,Monoclonal antibody ,magnetic beads ,Article ,Cost reduction ,Cricetulus ,Adsorption ,medicine ,Animals ,Batch cell culture ,process integration ,cell protein ,Host cell protein ,Bioprocess Technology ,Fed-batch cultures ,nonhuman ,cell suspension ,Clarification ,Magnetic Fields ,monoclonal antibody ,adsorption ,Biophysics ,Monoclonal antibodies ,Biokemi och molekylärbiologi ,cell density - Abstract
Cell clarification represents a major challenge for the intensification through very high cell density in the production of biopharmaceuticals such as monoclonal antibodies (mAbs). The present report proposes a solution to this challenge in a streamlined process where cell clarification and mAb capture are performed in a single step using magnetic beads coupled with protein A. Capture of mAb from non-clarified CHO cell suspension showed promising results; however, it has not been demonstrated that it can handle the challenge of very high cell density as observed in intensified fed-batch cultures. The performances of magnetic bead-based mAb capture on non-clarified cell suspension from intensified fed-batch culture were studied. Capture from a culture at density larger than 100 × 106 cells/ml provided an adsorption efficiency of 99% and an overall yield of 93% with a logarithmic host cell protein (HCP) clearance of ≈2–3 and a resulting HCP concentration ≤≈5 ppm. These results show that direct capture from very high cell density cell suspension is possible without prior processing. This technology, which brings significant benefits in terms of operational cost reduction and performance improvements such as low HCP, can be a powerful tool alleviating the challenge of process intensification. QC 20221102
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- 2021
35. A Novel Laser-Based Zebrafish Model for Studying Traumatic Brain Injury and Its Molecular Targets
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Tikhonova, M. A., Maslov, N. A., Bashirzade, A. A., Nehoroshev, E. V., Babchenko, V. Y., Chizhova, N. D., Tsibulskaya, E. O., Akopyan, A. A., Markova, E. V., Yang, Y. -L., Lu, K. -T., Kalueff, A. V., Aftanas, L. I., Amstislavskaya, T. G., Tikhonova, M. A., Maslov, N. A., Bashirzade, A. A., Nehoroshev, E. V., Babchenko, V. Y., Chizhova, N. D., Tsibulskaya, E. O., Akopyan, A. A., Markova, E. V., Yang, Y. -L., Lu, K. -T., Kalueff, A. V., Aftanas, L. I., and Amstislavskaya, T. G.
- Abstract
Traumatic brain injury (TBI) is a major public health problem. Here, we developed a novel model of non-invasive TBI induced by laser irradiation in the telencephalon of adult zebrafish (Danio rerio) and assessed their behavior and neuromorphology to validate the model and evaluate potential targets for neuroreparative treatment. Overall, TBI induced hypolocomotion and anxiety-like behavior in the novel tank test, strikingly recapitulating responses in mammalian TBI models, hence supporting the face validity of our model. NeuN-positive cell staining was markedly reduced one day, but not seven days, after TBI, suggesting increased neuronal damage immediately after the injury, and its fast recovery. The brain-derived neurotrophic factor (Bdnf) level in the brain dropped immediately after the trauma, but fully recovered seven days later. A marker of microglial activation, Iba1, was elevated in the TBI brain, albeit decreasing from Day 3. The levels of hypoxia-inducible factor 1-alpha (Hif1a) increased 30 min after the injury, and recovered by Day 7, further supporting the construct validity of the model. Collectively, these findings suggest that our model of laser-induced brain injury in zebrafish reproduces mild TBI and can be a useful tool for TBI research and preclinical neuroprotective drug screening. © 2022 by the authors.
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- 2022
36. Discovery of Nitro-azolo[1,5-a]pyrimidines with Anti-Inflammatory and Protective Activity against LPS-Induced Acute Lung Injury
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Spasov, A., Kosolapov, V., Babkov, D., Klochkov, V., Sokolova, E., Miroshnikov, M., Borisov, A., Velikorodnaya, Y., Smirnov, A., Savateev, K., Fedotov, V., Kotovskaya, S., Rusinov, V., Spasov, A., Kosolapov, V., Babkov, D., Klochkov, V., Sokolova, E., Miroshnikov, M., Borisov, A., Velikorodnaya, Y., Smirnov, A., Savateev, K., Fedotov, V., Kotovskaya, S., and Rusinov, V.
- Abstract
Acute lung injury remains a challenging clinical condition, necessitating the development of novel, safe and efficient treatments. The prevention of macrophage M1-polarization is a viable venue to tackle excessive inflammation. We performed a phenotypic screening campaign to identify azolopyrimidine compounds that effectively inhibit LPS-induced NO synthesis and interleukin 6 (IL-6) secretion. We identified lead compound 9g that inhibits IL-6 secretion with IC50 of 3.72 µM without apparent cytotoxicity and with minimal suppression of macrophage phagocytosis in contrast to dexamethasone. In a mouse model of LPS-induced acute lung injury, 30 mg/kg i.p. 9g ameliorated anxiety-like behavior, inhibited IL-6 release, and limited neutrophil infiltration and pulmonary edema. A histological study confirmed the protective activity of 9g. Treatment with compound 9g prevented the migration of CD68+ macrophages and the incidence of hemorrhage. Hence, we have identified a promising pharmacological approach for the treatment of acute lung injury that may hold promise for the development of novel drugs against cytokine-mediated complications of bacterial and viral infections. © 2022 by the authors. Licensee MDPI, Basel, Switzerland.
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- 2022
37. Age-dependent aneuploidy in mammalian oocytes instigated at the second meiotic division
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Kouznetsova, A., Liu, J. G., Valentiniene, S., Brismar, Hjalmar, Höög, C., Kouznetsova, A., Liu, J. G., Valentiniene, S., Brismar, Hjalmar, and Höög, C.
- Abstract
Ageing severely affects the chromosome segregation process in human oocytes resulting in aneuploidy, infertility and developmental disorders. A considerable amount of segregation errors in humans are introduced at the second meiotic division. We have here compared the chromosome segregation process in young adult and aged female mice during the second meiotic division. More than half of the oocytes in aged mice displayed chromosome segregation irregularities at anaphase II, resulting in dramatically increased level of aneuploidy in haploid gametes, from 4% in young adult mice to 30% in aged mice. We find that the post-metaphase II process that efficiently corrects aberrant kinetochore-microtubule attachments in oocytes in young adult mice is approximately 10-fold less efficient in aged mice, in particular affecting chromosomes that show small inter-centromere distances at the metaphase II stage in aged mice. Our results reveal that post-metaphase II processes have critical impact on age-dependent aneuploidy in mammalian eggs., QC 20230228
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- 2022
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38. An Adaptable Antibody-Based Platform for Flexible Synthetic Peptide Delivery Built on Agonistic CD40 Antibodies
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Eltahir, M., Laurén, I., Lord, M., Chourlia, A., Dahllund, Leif, Olsson, Anders, Saleh, A., Ytterberg, A. J., Lindqvist, A., Andersson, Oskar, Persson, Helena, Mangsbo, S. M., Eltahir, M., Laurén, I., Lord, M., Chourlia, A., Dahllund, Leif, Olsson, Anders, Saleh, A., Ytterberg, A. J., Lindqvist, A., Andersson, Oskar, Persson, Helena, and Mangsbo, S. M.
- Abstract
The agonistic potentials of therapeutic anti-CD40 antibodies have been profiled in relation to antibody isotype and epitope specificity. Still, clinical impact relies on a well-balanced clinical efficacy versus target-mediated toxicity. As CD40-mediated immune activation must rely on a combination of stimulation of antigen-presenting cells (APCs) alongside antigen presentation, for efficient T cell priming, alternative approaches to improve the therapeutic outcome of CD40-targeting strategies should focus on providing optimal antigen presentation together with CD40 stimulation. Herein, a bispecific antibody targeting CD40 as a means to deliver cargo (i.e., synthetic peptides) into APCs through a non-covalent, high-affinity interaction between the antibody and the cargo peptide, further referred to as the Adaptable Drug Affinity Conjugate (ADAC) technology, has been developed. The ADAC platform demonstrated a target-specific CD4+ and CD8+ T cell expansion in vitro and significantly improved peptide-specific CD8+ T cell proliferation in vivo. In addition, the strategy dramatically improved the in vitro and in vivo half-life of the synthetic peptides. Future applications of ADAC involve pandemic preparedness to viral genetic drift as well as neoepitope vaccination strategies where the bispecific antibody is an off-the-shelf product, and the peptide antigen is synthesized based on next-generation sequencing data mining., QC 20230308
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- 2022
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39. Cell transcriptomic atlas of the non-human primate Macaca fascicularis
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Han, L., Wei, X., Liu, C., Volpe, G., Zhuang, Z., Zou, X., Wang, Z., Pan, T., Yuan, Y., Zhang, X., Fan, P., Guo, P., Lai, Y., Lei, Y., Liu, X., Yu, F., Shangguan, S., Lai, G., Deng, Q., Liu, Y., Wu, L., Shi, Q., Yu, H., Huang, Y., Cheng, M., Xu, J., Wang, M., Wang, C., Zhang, Y., Xie, D., Yang, Y., Yu, Y., Zheng, H., Wei, Y., Huang, F., Lei, J., Huang, W., Zhu, Z., Lu, H., Wang, B., Chen, F., Yang, T., Du, W., Chen, J., Xu, S., An, J., Ward, C., Pei, Z., Wong, C. -W, Zhang, H., Liu, M., Qin, B., Schambach, A., Isern, J., Feng, L., Guo, X., Liu, Z., Sun, Q., Maxwell, P. H., Barker, N., Muñoz-Cánoves, P., Gu, Y., Mulder, Jan, Uhlén, Mathias, Tan, T., Liu, S., Yang, H., Wang, J., Hou, Y., Xu, X., Esteban, M. A., Liu, L., Han, L., Wei, X., Liu, C., Volpe, G., Zhuang, Z., Zou, X., Wang, Z., Pan, T., Yuan, Y., Zhang, X., Fan, P., Guo, P., Lai, Y., Lei, Y., Liu, X., Yu, F., Shangguan, S., Lai, G., Deng, Q., Liu, Y., Wu, L., Shi, Q., Yu, H., Huang, Y., Cheng, M., Xu, J., Wang, M., Wang, C., Zhang, Y., Xie, D., Yang, Y., Yu, Y., Zheng, H., Wei, Y., Huang, F., Lei, J., Huang, W., Zhu, Z., Lu, H., Wang, B., Chen, F., Yang, T., Du, W., Chen, J., Xu, S., An, J., Ward, C., Pei, Z., Wong, C. -W, Zhang, H., Liu, M., Qin, B., Schambach, A., Isern, J., Feng, L., Guo, X., Liu, Z., Sun, Q., Maxwell, P. H., Barker, N., Muñoz-Cánoves, P., Gu, Y., Mulder, Jan, Uhlén, Mathias, Tan, T., Liu, S., Yang, H., Wang, J., Hou, Y., Xu, X., Esteban, M. A., and Liu, L.
- Abstract
Studying tissue composition and function in non-human primates (NHPs) is crucial to understand the nature of our own species. Here we present a large-scale cell transcriptomic atlas that encompasses over 1 million cells from 45 tissues of the adult NHP Macaca fascicularis. This dataset provides a vast annotated resource to study a species phylogenetically close to humans. To demonstrate the utility of the atlas, we have reconstructed the cell–cell interaction networks that drive Wnt signalling across the body, mapped the distribution of receptors and co-receptors for viruses causing human infectious diseases, and intersected our data with human genetic disease orthologues to establish potential clinical associations. Our M. fascicularis cell atlas constitutes an essential reference for future studies in humans and NHPs., QC 20230116
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- 2022
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40. A novel nano delivery system targeting different stages of osteoclasts
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Zhang, B., Zhao, J., Yan, Hongji, Zhao, Y., Tian, H., Wang, C., Wang, R., Jin, J., Chen, Y., Yang, C., LI, C., Jiao, Y., Zheng, K., Zhu, F., Tian, W., Zhang, B., Zhao, J., Yan, Hongji, Zhao, Y., Tian, H., Wang, C., Wang, R., Jin, J., Chen, Y., Yang, C., LI, C., Jiao, Y., Zheng, K., Zhu, F., and Tian, W.
- Abstract
Osteoclast (OC) abnormalities represent osteoporosis's critical mechanism (OP). OCs undergo multiple processes that range from monocytic to functional. Different drugs target OCs at different developmental stages; however, almost no Suitable drug-targeted delivery systems exist. Therefore, we designed two dual-targeting nanoparticles to target OCs at different functional stages. Using the calcitonin gene-related peptide receptor (CGRPR), which OC precursors highly express, and specific TRAPpeptides screened in the bone resorption lacuna, where mature OCs function, respectively, two types of dual-targeted nanoparticles were constructed. Afterwards, nanoparticles were grafted with hyaluronic acid (HA), which specifically binds to CD44 on the surface of the OCs. In vivo and in vitro experiments show that both nanoparticles have noticeable targeting effects on OCs. This suggests that dual-targeting nanoparticles designed for different functional periods of OC can be well targeted to the corresponding OC, and further promote the more precise delivery of drugs used to treat OP., QC 20221214
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- 2022
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41. Clonal relations in the mouse brain revealed by single-cell and spatial transcriptomics
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Ratz, M., von Berlin, L., Larsson, Ludvig, Martin, M., Westholm, J. O., La Manno, G., Lundeberg, Joakim, Frisén, J., Ratz, M., von Berlin, L., Larsson, Ludvig, Martin, M., Westholm, J. O., La Manno, G., Lundeberg, Joakim, and Frisén, J.
- Abstract
The mammalian brain contains many specialized cells that develop from a thin sheet of neuroepithelial progenitor cells. Single-cell transcriptomics revealed hundreds of molecularly diverse cell types in the nervous system, but the lineage relationships between mature cell types and progenitor cells are not well understood. Here we show in vivo barcoding of early progenitors to simultaneously profile cell phenotypes and clonal relations in the mouse brain using single-cell and spatial transcriptomics. By reconstructing thousands of clones, we discovered fate-restricted progenitor cells in the mouse hippocampal neuroepithelium and show that microglia are derived from few primitive myeloid precursors that massively expand to generate widely dispersed progeny. We combined spatial transcriptomics with clonal barcoding and disentangled migration patterns of clonally related cells in densely labeled tissue sections. Our approach enables high-throughput dense reconstruction of cell phenotypes and clonal relations at the single-cell and tissue level in individual animals and provides an integrated approach for understanding tissue architecture., QC 20221109
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- 2022
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42. A Uniform and Isotropic Cytoskeletal Tiling Fills Dendritic Spines
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Eberhardt, F., Bushong, E. A., Phan, S., Peltier, S., Monteagudo-Mesas, P., Weinkauf, Tino, Herz, A. V. M., Stemmler, M., Ellisman, M., Eberhardt, F., Bushong, E. A., Phan, S., Peltier, S., Monteagudo-Mesas, P., Weinkauf, Tino, Herz, A. V. M., Stemmler, M., and Ellisman, M.
- Abstract
Dendritic spines are submicron, subcellular compartments whose shape is defined by actin filaments and associated proteins. Accurately mapping the cytoskeleton is a challenge, given the small size of its components. It remains unclear whether the actin-associated structures analyzed in dendritic spines of neurons in vitro apply to dendritic spines of intact, mature neurons in situ. Here, we combined advanced preparative methods with multitilt serial section electron microscopy (EM) tomography and computational analysis to reveal the full three-dimensional (3D) internal architecture of spines in the intact brains of male mice at nanometer resolution. We compared hippocampal (CA1) pyramidal cells and cerebellar Purkinje cells in terms of the length distribution and connectivity of filaments, their branching-angles and absolute orientations, and the elementary loops formed by the network. Despite differences in shape and size across spines and between spine heads and necks, the internal organization was remarkably similar in both neuron types and largely homogeneous throughout the spine volume. In the tortuous mesh of highly branched and interconnected filaments, branches exhibited no preferred orientation except in the immediate vicinity of the cell membrane. We found that new filaments preferentially split off from the convex side of a bending filament, consistent with the behavior of Arp2/3-mediated branching of actin under mechanical deformation. Based on the quantitative analysis, the spine cytoskeleton is likely subject to considerable mechanical force in situ., QC 20230613
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- 2022
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43. Development of L-Dopa-containing diketopiperazines as blood-brain barrier shuttle
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Cornacchia, C., Marinelli, L., Di Rienzo, A., Dimmito, M. P., Serra, F., Di Biase, G., De Filippis, B., Turkez, H., Mardinoglu, Adil, Bellezza, I., Di Stefano, A., Cacciatore, I., Cornacchia, C., Marinelli, L., Di Rienzo, A., Dimmito, M. P., Serra, F., Di Biase, G., De Filippis, B., Turkez, H., Mardinoglu, Adil, Bellezza, I., Di Stefano, A., and Cacciatore, I.
- Abstract
In our overall goal to develop anti-Parkinson drugs, we designed novel diketopiperazines (DKP1-6) aiming to both reach the blood-brain barrier and counteract the oxidative stress related to Parkinson's Disease (PD). The anti-Parkinson properties of DKP 1–6 were evaluated using neurotoxin-treated PC12 cells, as in vitro model of PD, while their cytotoxicity and genotoxicity potentials were investigated in newborn rat cerebral cortex (RCC) and primary human whole blood (PHWB) cell cultures. The response against free radicals was evaluated by the total antioxidant capacity (TAC) assay. Comet assay was used to detect DNA damage while the content of 8-hydroxyl-2′-deoxyguanosine (8-OH-dG) was determined as a marker of oxidative DNA damage. PAMPA-BBB and Caco-2 assays were employed to evaluate the capability of DKP1-6 to cross the membranes. Stability studies were conducted in simulated gastric and intestinal fluids and human plasma. Results showed that DKP5-6 attenuate the MPP + -induced cell death on a nanomolar scale, but a remarkable effect was observed for DKP6 on Nrf2 activation that leads to the expression of genes involved in oxidative stress response thus increasing glutathione biosynthesis and ROS buffering. DKP5-6 resulted in no toxicity for RCC neurons and PHWB cells exposed to 10–500 nM concentrations during 24 h as determined by MTT and LDH assays and TAC levels were not altered in both cultured cell types. No significant difference in the induction of DNA damage was observed for DKP5-6. Both DKPs resulted stable in simulated gastric fluids (t1/2 > 22h). In simulated intestinal fluids, DKP5 underwent immediate hydrolysis while DKP6 showed a half-life higher than 3 h. In human plasma, DKP6 resulted quite stable. DKP6 displayed both high BBB and Caco-2 permeability confirming that the DKP scaffold represents a useful tool to improve the crossing of drugs through the biological membranes., QC 20230613
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- 2022
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44. A universal SARS-CoV DNA vaccine inducing highly cross-reactive neutralizing antibodies and T cells
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Appelberg, S., Ahlén, G., Yan, J., Nikouyan, N., Weber, S., Larsson, O., Höglund, U., Aleman, S., Weber, F., Perlhamre, E., Apro, J., Gidlund, E. -K, Tuvesson, O., Salati, S., Cadossi, M., Tegel, Hanna, Hober, Sophia, Frelin, L., Mirazimi, A., Sällberg, M., Appelberg, S., Ahlén, G., Yan, J., Nikouyan, N., Weber, S., Larsson, O., Höglund, U., Aleman, S., Weber, F., Perlhamre, E., Apro, J., Gidlund, E. -K, Tuvesson, O., Salati, S., Cadossi, M., Tegel, Hanna, Hober, Sophia, Frelin, L., Mirazimi, A., and Sällberg, M.
- Abstract
New variants in the SARS-CoV-2 pandemic are more contagious (Alpha/Delta), evade neutralizing antibodies (Beta), or both (Omicron). This poses a challenge in vaccine development according to WHO. We designed a more universal SARS-CoV-2 DNA vaccine containing receptor-binding domain loops from the huCoV-19/WH01, the Alpha, and the Beta variants, combined with the membrane and nucleoproteins. The vaccine induced spike antibodies crossreactive between huCoV-19/WH01, Beta, and Delta spike proteins that neutralized huCoV-19/WH01, Beta, Delta, and Omicron virus in vitro. The vaccine primed nucleoprotein-specific T cells, unlike spike-specific T cells, recognized Bat-CoV sequences. The vaccine protected mice carrying the human ACE2 receptor against lethal infection with the SARS-CoV-2 Beta variant. Interestingly, priming of cross-reactive nucleoprotein-specific T cells alone was 60% protective, verifying observations from humans that T cells protect against lethal disease. This SARS-CoV vaccine induces a uniquely broad and functional immunity that adds to currently used vaccines., QC 20230523
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- 2022
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45. NucleoCounter – An Efficient Technique for the Determination of Cell Number and Viability in Animal Cell Culture Processes
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Shah, Dimpalkumar, Clee, Paul, Boisen, Sthen, Al-Rubeai, Mohammed, and Smith, Rodney, editor
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- 2007
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46. Bigger Cells : Eukaryotic cells and their contents
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Agutter, Paul S., Wheatley, Denys N., Agutter, Paul S., and Wheatley, Denys N.
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- 2007
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47. Cell-Cell Transport of Homeoproteins : With or Without Channels?
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Joliot, Alain, Prochiantz, Alain, Baluska, Frantisek, Volkmann, Dieter, and Barlow, Peter W.
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- 2006
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48. The effect of aromatase inhibitors against possible testis toxicity in pembrolizumab treated rats
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Neşe Başak Türkmen, Osman Çiftçi, Aslı Taşlıdere, Muhterem Aydın, and Binay Can Eke
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reproductive toxicity ,Male ,antioxidant ,sperm quality ,Programmed Cell Death 1 Receptor ,animal cell ,Antioxidants ,Rats, Sprague-Dawley ,Endocrinology ,Testis ,rat ,animal ,Testosterone ,Sprague Dawley rat ,Aromatase Inhibitors ,adult ,General Medicine ,testosterone blood level ,Catalase ,Glutathione ,histopathology ,Sperm Motility ,pembrolizumab ,testis weight ,epididymis ,seminiferous tubule ,Urology ,thiobarbituric acid reactive substance ,animal experiment ,testis tissue ,Anastrozole ,Antibodies, Monoclonal, Humanized ,sperm ,Thiobarbituric Acid Reactive Substances ,Article ,semen analysis ,animal tissue ,histology ,Aromatase ,Semen ,Animals ,controlled study ,spermatozoon density ,Glutathione Peroxidase ,nonhuman ,Superoxide Dismutase ,animal model ,spermatozoon motility ,Polyphenols ,programmed death 1 receptor ,therapy effect ,Rats ,polyphenol ,monoclonal antibody ,Resveratrol ,testis disease ,aromatase inhibitor - Abstract
Pembrolizumab is a monoclonal antibody. Anastrozole is an infertility inhibitor of aromatase. Resveratrol is an antioxidant polyphenol in the reproductive system. This study was planned to demonstrate the protective effects of anastrozole and resveratrol against pembrolizumab-induced reproductive damage. Forty-two Sprague–Dawley rats were used in the study. Groups: The control, Pembrolizumab (PEMB), PEMB + Anastrazol (ANAST), PEMB + Resveratrol (RES), RES, and ANAST groups. At the end of the experiment, rats were euthanased under anaesthesia. Tissue samples were taken from rats for biochemical, histological, and ELISA evaluations. Tissues were subjected to routine tissue follow-up for histological analysis. Biochemically, thiobarbituric acid reactive substance (TBARS), glutathione (GSH), catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) levels were measured. Sperm motility, abnormal sperm rate, and epididymal sperm concentration were examined spermatologically. Serum testosterone and programmed cell death-1 (PD-1) levels were measured using the ELISA. TBARS levels were significantly increased and GSH, SOD, GPx, and CAT levels were mitigated in PEMB-treated rats. Histologically; Control, ANAST, and RES groups testis samples were observed with normal histological appearance. Histological damage was detected in seminiferous tubule structures in testicular tissue in the PEMB group. In treatment groups, this damage was decreased. In addition, PD-1 and testosterone levels were evaluated by the ELISA method. ANAST and RES have therapeutic effects against reproductive damage caused by PEMB. © 2022 Wiley-VCH GmbH.
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- 2022
49. Target-Driven Design of a Coumarinyl Chalcone Scaffold Based Novel EF2 Kinase Inhibitor Suppresses Breast Cancer Growth In Vivo
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Serdar Durdagi, Esen Bellur Atici, Tugba Taskin Tok, Mehmet Ay, Bulent Ozpolat, Bekir Karliga, Goknur Kara, Gizem Tatar, Hakan Kandemir, Ferah Cömert Önder, Nermin Kahraman, and Ali Cagir
- Subjects
BT-20 cell line ,protein p53 ,medicine.medical_treatment ,blood brain barrier ,animal cell ,MCF-7 cell line ,coumarin ,Western blotting ,Targeted therapy ,lipophilicity ,Pharmacology (medical) ,enzyme phosphorylation ,cancer survival ,liposomal delivery ,education.field_of_study ,Chemistry ,Kinase ,nanoparticle ,quantum mechanics ,apoptosis ,MDA-MB-231 cell line ,EF2K ,female ,priority journal ,molecularly targeted therapy ,enzyme active site ,crystal structure ,in vitro study ,pharmacokinetic parameters ,drug design ,animal experiment ,cardiotoxicity ,chalcone ,phosphotransferase inhibitor ,densitometry ,colony formation ,Article ,cancer growth ,in vivo study ,breast cancer ,Breast cancer ,In vivo ,medicine ,controlled study ,time-dependent inhibition ,education ,mouse ,Pharmacology ,MDA-MB-436 cell line ,nonhuman ,molecular modeling ,Cell growth ,animal model ,molecular docking ,medicine.disease ,elongation factor 2 kinase ,molecular dynamics ,tumor xenograft ,In vitro ,Elongation factor ,cell proliferation ,Cancer research ,protein kinase B ,drug synthesis ,imidazole derivative ,Elongation Factor-2 Kinase - Abstract
Eukaryotic elongation factor 2 kinase (eEF-2K) is an unusual alpha kinase involved in protein synthesis through phosphorylation of elongation factor 2 (EF2). eEF-2K is highly overexpressed in breast cancer, and its activity is associated with significantly shortened patient survival and proven to be a potential molecular target in breast cancer. The crystal structure of eEF-2K remains unknown, and there is no potent, safe, and effective inhibitor available for clinical applications. We designed and synthesized several generations of potential inhibitors. The effect of the inhibitors at the binding pocket of eEF-2K was analyzed after developing a 3D target model by using a domain of another ?-kinase called myosin heavy-chain kinase A (MHCKA) that closely resembles eEF-2K. In silico studies showed that compounds with a coumarin-chalcone core have high predicted binding affinities for eEF-2K. Using in vitro studies in highly aggressive and invasive (MDA-MB-436, MDA-MB-231, and BT20) and noninvazive (MCF-7) breast cancer cells, we identified a lead compound that was highly effective in inhibiting eEF-2K activity at submicromolar concentrations and at inhibiting cell proliferation by induction of apoptosis with no toxicity in normal breast epithelial cells. In vivo systemic administration of the lead compound encapsulated in single lipid-based liposomal nanoparticles twice a week significantly suppressed growth of MDA-MB-231 tumors in orthotopic breast cancer models in nude mice with no observed toxicity. In conclusion, our study provides a highly potent and in vivo effective novel small-molecule eEF-2K inhibitor that may be used as a molecularly targeted therapy breast cancer or other eEF-2K-dependent tumors. © 2021 American Chemical Society. 1R01CA244344; University of Texas MD Anderson Cancer Center; Türkiye Bilimsel ve Teknolojik Araştirma Kurumu, TÜBITAK: 215S008, TUBITAK-BIDEB 2214A This study was funded by The Scientific and Technological Research Council of Turkey (TUBITAK) (grant number 215S008 and TUBITAK-BIDEB 2214A program, F.C.O.) and The University of Texas-MD Anderson Cancer Center Bridge fund (B.O. and N.K.) and NIH-NCI 1R01CA244344 grants (B.O. and N.K.).
- Published
- 2021
50. The Viable Cell Monitor: A Dielectric spectroscope for Growth and Metabolic Studies of Animal Cells on Macroporous Beads
- Author
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Guan, Y., Kemp, R. B., Merten, Otto-Wilhelm, editor, Perrin, Pierre, editor, and Griffiths, Bryan, editor
- Published
- 2002
- Full Text
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