The re-emerging disease histomonosis is caused by the protozoan parasite Histomonas meleagridis that affects chickens and turkeys. Previously, protection by vaccination with in vitro attenuated H. meleagridis has been demonstrated and an involvement of T cells, potentially by IFN-γ production, was hypothesized. However, comparative studies between chickens and turkeys on H. meleagridis-specific T cells were not conducted yet. This work investigated IFN-γ production within CD4+, CD8α+ and TCRγδ+ (chicken) or CD3ε+CD4-CD8α- (turkey) T cells of spleen and liver from vaccinated and/or infected birds using clonal cultures of a monoxenic H. meleagridis strain. In infected chickens, re-stimulated splenocytes showed a significant increase of IFN-γ+CD4+ T cells. Contrariwise, significant increments of IFN-γ-producing cells within all major T-cell subsets of the spleen and liver were found for vaccinated/infected turkeys. This indicates that the vaccine in turkeys causes more intense systemic immune responses whereas in chickens protection might be mainly driven by local immunity.The four-and-a-half LIM-only protein family of transcription co-factors participates in various cellular processes, such as cell proliferation, cell differentiation, apoptosis, cell adhesion, migration, transcription and signal transduction. However, the knowledge of the structural characteristics and immune functions of its ancestor Lmpt, which contains six LIM domains at the C-terminus and a PET domain at the N-terminus, is limited in invertebrates, especially in crustaceans. In the present study, a novel Lmpt from oriental river prawn (Macrobrachium nipponense) was identified, and its role in the immune response was investigated. Its full-length cDNA sequence was 6407 bp, which contained a 2595 bp ORF encoding 865 amino acids, exhibiting high similarity to the structure of Lmpt derived from other invertebrates. Tissue distribution analysis revealed that MnLmpt was widely expressed in all examined tissues, and high expression levels were observed in muscle, heart and intestine in M. https://www.selleckchem.com/ nipponense. After experiese results showed that MnLmpt might play a crucial role in the innate immune response in M. nipponense, and these findings paved the way for a better understanding of the immune system in crustacean species.TNK1 (thirty-eight-negative kinase 1) belongs to the ACK (Activated Cdc42 Kinases) family of intracellular non-receptor tyrosine kinases that usually acts as an important regulator in cytokine receptor-mediated intracellular signal transduction pathways. JAK-STAT signal pathway acts as a key point in cellular proliferation, differentiation and immunomodulatory. Mammalian TNK1 is involved in antiviral immunity and activation of growth factors. However, TNK1 has rarely been studied in fish. To evaluate the role of fish TNK1 in JAK-STAT pathway, we cloned the full-length cDNA sequence of grass carp (Ctenopharyngodon idella) TNK1 (CiTNK1). CiTNK1 protein consists of N-terminal Tyrkc (tyrosine kinase) domain, C-terminal SH3 (Src homology 3) domain and Pro-rich domain. Phylogenetic analysis showed that CiTNK1 has a closer relationship with Danio rerio TNK1. The expression and phosphorylation of CiTNK1 in grass carp tissues and cells was increased under poly(IC) stimulation. Subcellular localization and co-immunoprecipitation indicated that CiTNK1 is targeted in the cytoplasm and interacts with grass carp STAT1 (CiSTAT1). Co-transfection of CiTNK1 and CiSTAT1 into cells facilitated the expression of IFN I. This is because that the presence of CiTNK1 enhanced the phosphorylation of CiSTAT1 and causes activation of CiSTAT1. Our results revealed that TNK1 can potentiate the phosphorylation of STAT1 and then regulates JAK-STAT pathway to trigger IFN I expression in fish.BELAVY, D. L., J. Van Oosterwijck, M. Clarkson, E. Dhondt, N. L. Mundell, C. Miller and P. J. Owen. NEUROSCI BIOBEHAV REV 21(1) XXX-XXX, 2020. Exercise training is capable of reducing pain in chronic pain syndromes, yet its mechanisms are less well established. One mechanism may be via the impact of exercise on increasing a person's pain threshold. Here we show, via meta-analysis of fifteen exercise training studies in pain syndromes that exercise training leads to increased pressure pain thresholds (low to moderate quality evidence). We also find low to moderate quality evidence exists that exercise training was more effective than non-exercise interventions, such as pain education, massage and stress management for improving pain sensitivity. Further, the effect of exercise was greater locally at the site of pain and less so at remote regions. These finding suggest that adaptations in central inhibition occur over time with exercise training and, more widely, add to the mechanistic understanding of how effective interventions can improve pain in chronic pain syndromes.The multitude of clinical trials using mesenchymal stromal cells (****) has underscored their significance as a promising cell source for regenerative therapies. Most studies have however shown that **** get entrapped into the microvasculature of lungs, liver and spleen. In addition to intercellular communication, **** exert their effects in a paracrine manner by secretion of extracellular vesicles (EVs). The therapeutic effects of ****derived EVs have been examined in several diseases such as hepatic failure, liver injury, hematopoiesis etc. Therefore, optimization of cryopreservation strategies for the long-term storage of functional EVs could help in the development of off-the-shelf biologics. The aim of this study was to develop an optimal cryopreservation strategy for the efficient storage of both types of EVs - Microvesicles (MVs) and exosomes, independently, and to further examine the effect of the cryopreserved EVs on the ex vivo expansion of HSCs. MVs and exosomes were separately cryopreserved at different temperatures using PBS or PBS supplemented with trehalose (pTRE), and these cryopreserved EVs were then assessed for their functionality after revival. We found that addition of trehalose during cryopreservation helped in maintaining the morphology and functionality of the EVs, as assessed by their HSC-supportive potential, ability to expand phenotypically defined HSCs and ability to maintain the chemotactic migration potential of the HSCs co-cultured with them. This strategy could prove to be beneficial for facilitating the use of EVs as cell-free ready-to-use biologics for the ex vivo expansion of HSCs and in regenerative medicine.
The re-emerging disease histomonosis is caused by the protozoan parasite Histomonas meleagridis that affects chickens and turkeys. Previously, protection by vaccination with in vitro attenuated H. meleagridis has been demonstrated and an involvement of T cells, potentially by IFN-γ production, was hypothesized. However, comparative studies between chickens and turkeys on H. meleagridis-specific T cells were not conducted yet. This work investigated IFN-γ production within CD4+, CD8α+ and TCRγδ+ (chicken) or CD3ε+CD4-CD8α- (turkey) T cells of spleen and liver from vaccinated and/or infected birds using clonal cultures of a monoxenic H. meleagridis strain. In infected chickens, re-stimulated splenocytes showed a significant increase of IFN-γ+CD4+ T cells. Contrariwise, significant increments of IFN-γ-producing cells within all major T-cell subsets of the spleen and liver were found for vaccinated/infected turkeys. This indicates that the vaccine in turkeys causes more intense systemic immune responses whereas in chickens protection might be mainly driven by local immunity.The four-and-a-half LIM-only protein family of transcription co-factors participates in various cellular processes, such as cell proliferation, cell differentiation, apoptosis, cell adhesion, migration, transcription and signal transduction. However, the knowledge of the structural characteristics and immune functions of its ancestor Lmpt, which contains six LIM domains at the C-terminus and a PET domain at the N-terminus, is limited in invertebrates, especially in crustaceans. In the present study, a novel Lmpt from oriental river prawn (Macrobrachium nipponense) was identified, and its role in the immune response was investigated. Its full-length cDNA sequence was 6407 bp, which contained a 2595 bp ORF encoding 865 amino acids, exhibiting high similarity to the structure of Lmpt derived from other invertebrates. Tissue distribution analysis revealed that MnLmpt was widely expressed in all examined tissues, and high expression levels were observed in muscle, heart and intestine in M. https://www.selleckchem.com/ nipponense. After experiese results showed that MnLmpt might play a crucial role in the innate immune response in M. nipponense, and these findings paved the way for a better understanding of the immune system in crustacean species.TNK1 (thirty-eight-negative kinase 1) belongs to the ACK (Activated Cdc42 Kinases) family of intracellular non-receptor tyrosine kinases that usually acts as an important regulator in cytokine receptor-mediated intracellular signal transduction pathways. JAK-STAT signal pathway acts as a key point in cellular proliferation, differentiation and immunomodulatory. Mammalian TNK1 is involved in antiviral immunity and activation of growth factors. However, TNK1 has rarely been studied in fish. To evaluate the role of fish TNK1 in JAK-STAT pathway, we cloned the full-length cDNA sequence of grass carp (Ctenopharyngodon idella) TNK1 (CiTNK1). CiTNK1 protein consists of N-terminal Tyrkc (tyrosine kinase) domain, C-terminal SH3 (Src homology 3) domain and Pro-rich domain. Phylogenetic analysis showed that CiTNK1 has a closer relationship with Danio rerio TNK1. The expression and phosphorylation of CiTNK1 in grass carp tissues and cells was increased under poly(IC) stimulation. Subcellular localization and co-immunoprecipitation indicated that CiTNK1 is targeted in the cytoplasm and interacts with grass carp STAT1 (CiSTAT1). Co-transfection of CiTNK1 and CiSTAT1 into cells facilitated the expression of IFN I. This is because that the presence of CiTNK1 enhanced the phosphorylation of CiSTAT1 and causes activation of CiSTAT1. Our results revealed that TNK1 can potentiate the phosphorylation of STAT1 and then regulates JAK-STAT pathway to trigger IFN I expression in fish.BELAVY, D. L., J. Van Oosterwijck, M. Clarkson, E. Dhondt, N. L. Mundell, C. Miller and P. J. Owen. NEUROSCI BIOBEHAV REV 21(1) XXX-XXX, 2020. Exercise training is capable of reducing pain in chronic pain syndromes, yet its mechanisms are less well established. One mechanism may be via the impact of exercise on increasing a person's pain threshold. Here we show, via meta-analysis of fifteen exercise training studies in pain syndromes that exercise training leads to increased pressure pain thresholds (low to moderate quality evidence). We also find low to moderate quality evidence exists that exercise training was more effective than non-exercise interventions, such as pain education, massage and stress management for improving pain sensitivity. Further, the effect of exercise was greater locally at the site of pain and less so at remote regions. These finding suggest that adaptations in central inhibition occur over time with exercise training and, more widely, add to the mechanistic understanding of how effective interventions can improve pain in chronic pain syndromes.The multitude of clinical trials using mesenchymal stromal cells (MSCs) has underscored their significance as a promising cell source for regenerative therapies. Most studies have however shown that MSCs get entrapped into the microvasculature of lungs, liver and spleen. In addition to intercellular communication, MSCs exert their effects in a paracrine manner by secretion of extracellular vesicles (EVs). The therapeutic effects of MSC-derived EVs have been examined in several diseases such as hepatic failure, liver injury, hematopoiesis etc. Therefore, optimization of cryopreservation strategies for the long-term storage of functional EVs could help in the development of off-the-shelf biologics. The aim of this study was to develop an optimal cryopreservation strategy for the efficient storage of both types of EVs - Microvesicles (MVs) and exosomes, independently, and to further examine the effect of the cryopreserved EVs on the ex vivo expansion of HSCs. MVs and exosomes were separately cryopreserved at different temperatures using PBS or PBS supplemented with trehalose (pTRE), and these cryopreserved EVs were then assessed for their functionality after revival. We found that addition of trehalose during cryopreservation helped in maintaining the morphology and functionality of the EVs, as assessed by their HSC-supportive potential, ability to expand phenotypically defined HSCs and ability to maintain the chemotactic migration potential of the HSCs co-cultured with them. This strategy could prove to be beneficial for facilitating the use of EVs as cell-free ready-to-use biologics for the ex vivo expansion of HSCs and in regenerative medicine.
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