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To develop and identify the effectiveness of a hospital-to-home transitional intervention based on an interaction model of client health behavior in adult patients with stroke. A non-equivalent control group pretest-posttest. Thirty-eight patients participated (18=intervention, 20=control); the intervention group received 12 weeks of the intervention. The intervention influenced anxiety, disease severity, health behavior adherence, patient satisfaction, and quality of life in adult patients with stroke. Transitional programs have potential to improve the health behaviors of subjects, and community health nurses can assist in the implementation of these programs. Health behaviors and quality of life scores were significantly higher in the intervention group than in the control group; these findings support the need for continuity of nursing care during the transitional period for patients with stroke. Given the challenges faced by adult stroke patients after stroke, community nurses should pay attention to patients' transitional experiences.
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Acidente Vascular Cerebral , Cuidado Transicional , Humanos , Adulto , Qualidade de Vida , Acidente Vascular Cerebral/terapia , Comportamentos Relacionados com a Saúde , Satisfação do Paciente , HospitaisRESUMO
NAD(P)-dependent steroid dehydrogenase-like (NSDHL), an essential enzyme in human cholesterol synthesis and a regulator of epidermal growth factor receptor (EGFR) trafficking pathways, has attracted interest as a therapeutic target due to its crucial relevance to cholesterol-related diseases and carcinomas. However, the development of pharmacological agents for targeting NSDHL has been hindered by the absence of the atomic details of NSDHL. In this study, we reported two X-ray crystal structures of human NSDHL, which revealed a detailed description of the coenzyme-binding site and the unique conformational change upon the binding of a coenzyme. A structure-based virtual screening and biochemical evaluation were performed and identified a novel inhibitor for NSDHL harboring suppressive activity towards EGFR. In EGFR-driven human cancer cells, treatment with the potent NSDHL inhibitor enhanced the antitumor effect of an EGFR kinase inhibitor. Overall, these findings could serve as good platforms for the development of therapeutic agents against NSDHL-related diseases.
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3-Hidroxiesteroide Desidrogenases/metabolismo , Inibidores Enzimáticos/metabolismo , 3-Hidroxiesteroide Desidrogenases/antagonistas & inibidores , 3-Hidroxiesteroide Desidrogenases/química , 3-Hidroxiesteroide Desidrogenases/genética , Sítios de Ligação , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Colesterol/química , Cristalografia por Raios X , Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , Receptores ErbB/antagonistas & inibidores , Receptores ErbB/metabolismo , Cloridrato de Erlotinib/química , Cloridrato de Erlotinib/metabolismo , Cloridrato de Erlotinib/farmacologia , Humanos , Cinética , Simulação de Acoplamento Molecular , Mutagênese Sítio-Dirigida , NAD/química , NAD/metabolismo , Estrutura Terciária de Proteína , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/isolamento & purificação , Transdução de SinaisRESUMO
The rhLIF is widely used as an essential factor in stem cell cultures for cell therapies. However, all the recombinant LIFs commercially available are expensive, and no commercially available rhLIF meet the standards recommended by USP for use in cell therapies. The current study reports the efficient production of N-glycosylated and bioactive rhLIF in CHO cells. The production rate of established rhLIF-expressing rCHO cells was approximately 0.85 g/l in 12-day fed-batch cultures using a 7.5 l bioreactor. The rhLIF protein was purified via a four-step purification procedure with approximately 57% recovery rate and greater than 99% purity. The purified rhLIF was N-glycosylated and biologically active with an EC50 of 0.167 ng/ml and a specific activity of 0.86 × 103 IU/mg. The purification procedure controlled the levels of process-related impurities below critical levels recommended by USP for cytokines used in cell therapies. The current work is the first production process of N-glycosylated and bioactive rhLIF, which can be applied to large-scale manufacture of GMP-grade rhLIF for use as an ancillary material in cell therapy.
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Fator Inibidor de Leucemia , Animais , Células CHO , Cricetulus , Glicosilação , Humanos , Fator Inibidor de Leucemia/biossíntese , Fator Inibidor de Leucemia/química , Fator Inibidor de Leucemia/genética , Fator Inibidor de Leucemia/isolamento & purificação , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificaçãoRESUMO
This study was conducted to develop and test a prediction model that explains health-related quality of life (HRQoL) of patients with Parkinson's disease (PD). Participants were 248 patients with PD enrolled in the neurology clinic of a university hospital in Seoul, Korea. The data were collected through structured questionnaires from March 1 to July 5, 2013. Motor fluctuations, depression, sleep disturbances, fatigue, and activities of daily living had significant direct effects on the quality of life of PD patients. Disease severity, social support, pain, sleep disturbances, fatigue, and activities of daily living had significant indirect effects on patients' quality of life. These predictive variables explained 77.4% of the total variance. The assessment of HRQoL in PD should be considered along with the variables affecting it, including social support. In addition, strategies to enhance social support may be useful for improving the quality of life for patients with PD.
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Nível de Saúde , Doença de Parkinson/psicologia , Qualidade de Vida/psicologia , Apoio Social , Fadiga , Feminino , Humanos , Masculino , Inquéritos e QuestionáriosRESUMO
We compared the nuclear maturation status and gene-expression profiles of canine cumulus cells (CCs) derived from cumulus-oocyte complexes (COCs) that were spontaneously ovulated versus those that were matured in vitro. Cumulus-oocyte complexes were retrieved from uteri by surgical flushing (after spontaneous ovulation) or by ovariectomy follicle aspiration and in vitro maturation. The objective of Experiment 1 was to investigate the nuclear maturation status of in vivo- versus in vitro-matured oocytes. The objective of Experiment 2 was to compare gene-expression profiles of CCs derived from in vivo- versus in vitro-matured COCs. Genes analysed are related to cell maturation, development and apoptosis, including GDF9, MAPK1, PTX3, CX43, Bcl2 and BAX; mRNA expression for all of these genes, except for GDF9, differed (P<0.05) between in vivo- and in vitro-matured CCs. In conclusion, we found that gene-expression profiles are related to the quality of CCs and therefore posit that monitoring gene expression could be a useful strategy to guide attempts to improve in vitro culture systems.
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Células do Cúmulo/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Técnicas de Maturação in Vitro de Oócitos , Oócitos/metabolismo , Oogênese , Ovulação , Animais , Cães , Feminino , Perfilação da Expressão Gênica/métodos , RNA Mensageiro/genética , RNA Mensageiro/metabolismoRESUMO
PURPOSE: This study attempted to estimate the utility weights for hypothetical chronic obstructive pulmonary disease (COPD) health states, including the effect of exacerbation, and based on utilities elicited from a representative sample using the time trade-off (TTO). METHODS: A total of 200 study subjects were recruited using a quota sampling method in Seoul, Korea. Ten COPD health profiles were described reflecting the severity of COPD and the extent of exacerbation. Respondents evaluated each health state using a visual analogue scale and TTO during a personal interview. TTO values were estimated using a linear mixed model, and the model performance was evaluated in terms of its predictive ability and goodness of fit. RESULTS: The estimated TTO values were 0.824 in moderate, 0.646 in severe, and 0.305 in very severe COPD health states. The estimated utility decrements in TTO varied from 0.082 for a non-serious exacerbation to 0.228 for one non-serious plus one serious exacerbation per year. The mean absolute error of the TTO model was 0.008, and the generalized R(2) was 0.86. CONCLUSION: The social preference of various COPD health states and the utility decrement due to exacerbation can be useful for the economic evaluation of COPD intervention in Korea.
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Pessoal de Saúde/estatística & dados numéricos , Nível de Saúde , Vigilância da População/métodos , Doença Pulmonar Obstrutiva Crônica/diagnóstico , Doença Pulmonar Obstrutiva Crônica/epidemiologia , Adulto , Idoso , Feminino , Humanos , Modelos Lineares , Masculino , Pessoa de Meia-Idade , Qualidade de Vida , República da CoreiaRESUMO
Hypertension is one of the most serious health problems in Korea. The purpose of this paper is to identify factors associated with self-reported nonadherence to an antihypertensive regimen. The data were obtained from the Korean Medical Panel 2008 and 2009 database, which were surveyed by the Korea Institute for Health and Social Affairs and National Health Insurance. We analyzed 5324 patients using multivariate logistic regression models. Self-reported nonadherence was used as a dependent variable and demographic, socioeconomic, and health status were included as independent variables to investigate the associated factors. Among the patients who were diagnosed with hypertension, 13.2% did not take their medicine as prescribed. Age and education attainment level were statistically significant. Younger and lower educational attainment groups were less likely to adhere to medication regimens and showed a tendency not to take their medicine as prescribed. There were no statistically significant variables in terms of health status. Our findings suggest that nurses in clinical and community settings should pay more attention to hypertensive patients who are young and less educated.
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Anti-Hipertensivos/administração & dosagem , Hipertensão/tratamento farmacológico , Adesão à Medicação , Adulto , Fatores Etários , Idoso , Demografia , Escolaridade , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , República da Coreia , Fatores de RiscoRESUMO
This study proposes a titanium silicide (TiSi2) recombination layer for perovskite/tunnel oxide passivated contact (TOPCon) 2-T tandem solar cells as an alternative to conventional transparent conductive oxide (TCO)-based recombination layers. TiSi2 was formed while TiO2 was made by oxidizing a Ti film deposited on the p+-Si layer. The reaction formation mechanism was proposed based on the diffusion theory supported by experimental results. The optical and electrical properties of the TiSi2 layer were optimized by controlling the initial Ti thicknesses (5-100 nm). With the initial Ti of 50 nm, the lowest reflectance and highly ohmic contact between the TiO2 and p+-Si layers with a contact resistivity of 161.48 mΩ·cm2 were obtained. In contrast, the TCO interlayer shows Schottky behavior with much higher contact resistivities. As the recombination layer of TiSi2 and the electron transport layer of TiO2 are formed simultaneously, the process steps become simpler. Finally, the MAPbI3/TOPCon tandem device yielded an efficiency of 16.23%, marking the first reported efficiency for a device including a silicide-based interlayer.
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Organ-on-a-chip (OOC) is an emerging interdisciplinary technology that reconstitutes the structure, function, and physiology of human tissues as an alternative to conventional preclinical models for drug screening. Over the last decade, substantial progress has been made in mimicking tissue- and organ-level functions on chips through technical advances in biomaterials, stem cell engineering, microengineering, and microfluidic technologies. Structural and engineering constituents, as well as biological components, are critical factors to be considered to reconstitute the tissue function and microenvironment on chips. In this review, we highlight critical engineering technologies for reconstructing the tissue microarchitecture and dynamic spatiotemporal microenvironment in OOCs. We review the technological advances in the field of OOCs for a range of applications, including systemic analysis tools that can be integrated with OOCs, multiorgan-on-chips, and large-scale manufacturing. We then discuss the challenges and future directions for the development of advanced end-user-friendly OOC systems for a wide range of applications.
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This systematic review and meta-analysis aimed to evaluate the efficacy and safety of acupuncture in treating emotional problems in women with infertility. We searched for randomized controlled trials using acupuncture treatment for emotional problems in women with infertility using 11 databases from their inception to 30 June 2023. The control intervention included no treatment, sham acupuncture treatment, and conventional treatment. The primary outcome was emotion-related rating scales, and the secondary outcomes were total effectiveness rate, quality of life, clinical pregnancy rate, and adverse events. Twelve randomized controlled trials involving 1930 participants were included. A meta-analysis of these studies indicated that, as compared to the control treatment, acupuncture significantly improved the State-Trait Anxiety Inventory, Self-rating Anxiety Scale, Amsterdam Preoperative Anxiety and Information Scale, and Self-rating Depression Scale scores, which were the primary emotion-related outcomes. Furthermore, the meta-analysis demonstrated that acupuncture treatment had a significant effect on the clinical pregnancy rate, which was the secondary outcome. No adverse events were reported in any of the studies. Our findings demonstrate the potential of acupuncture for treating emotional problems in women with infertility. However, well-designed and high-quality randomized clinical trials are required to confirm the effectiveness and safety of acupuncture treatment. The protocol of the current study was registered in PROSPERO (registration number: CRD42020166119).
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Mothers in the postpartum period often experience musculoskeletal disorders and pain, impacting their ability to care for themselves and their infants. Conventional treatments have limitations, prompting interest in alternative options like herbal medicine. This systematic review aimed to confirm the effectiveness and safety of herbal medicine treatment to improve maternal health in patients with postpartum pain (puerperal wind syndrome). We searched eight electronic databases for randomized controlled trials (RCTs) to evaluate the effects of herbal medicines on puerperal wind syndrome. Nine RCTs, including 652 patients, were selected. Following a meta-analysis of RCTs, both herbal medicine and combination treatments improved the visual analog scale scores, total effective rate, scores of Traditional Chinese Medicine syndromes, Oswestry Disability Index, and quality of life in patients with role-emotional puerperal wind syndrome. All adverse events were minor, and the incidence rate was not high compared with that of the control group. In conclusion, herbal medicine supports the improvement in pain, other systemic symptoms, and the quality of life of patients with puerperal wind syndrome. Moreover, no serious side effects were observed; therefore, herbal medicines appear to be safe. It can be the preferred treatment option for puerperal wind syndrome, which is currently managed symptomatically.
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OBJECTIVES: The purpose of this study was to explore new ways of creating value in the medical field and to derive recommendations for the role of medical institutions and the government. METHODS: In this paper, based on expert discussion, we classified Internet of Things (IoT) technologies into four categories according to the type of information they collect (location, environmental parameters, energy consumption, and biometrics), and investigated examples of application. RESULTS: Biometric IoT diagnoses diseases accurately and offers appropriate and effective treatment. Environmental parameter measurement plays an important role in accurately identifying and controlling environmental factors that could be harmful to patients. The use of energy measurement and location tracking technology enabled optimal allocation of limited hospital resources and increased the efficiency of energy consumption. The resulting economic value has returned to patients, improving hospitals' cost-effectiveness. CONCLUSIONS: Introducing IoT-based technology to clinical sites, including medical institutions, will enhance the quality of medical services, increase patient safety, improve management efficiency, and promote patient-centered medical services. Moreover, the IoT is expected to play an active role in the five major tasks of facility hygiene in medical fields, which are all required to deal with the COVID-19 pandemic: social distancing, contact tracking, bed occupancy control, and air quality management. Ultimately, the IoT is expected to serve as a key element for hospitals to perform their original functions more effectively. Continuing investments, deregulation policies, information protection, and IT standardization activities should be carried out more actively for the IoT to fulfill its expectations.
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PURPOSE: This study evaluated the effectiveness of the Self-Efficacy Reinforcement Stretching Exercise Program to improve osteoarthritis (OA)-related symptoms in older women with OA. DESIGN: A quasi-experimental design was used in this study. METHODS: The experimental group participated in the Self-Efficacy Reinforcement Stretching Exercise Program, a 6-week program composed of stretching exercises and self-efficacy reinforcement strategies, whereas the control group did not. All participants were assessed on pain; joint stiffness; physical function disability; body mass index; depression; and quality of life at the beginning, end, and 4 weeks after the program ended. FINDINGS: After participation, the experimental group had significantly less joint stiffness, physical function disability, and depression scores and significantly higher self-efficacy and quality of life than the control group. CONCLUSIONS: This intervention program led to an improvement in OA-related symptoms, self-efficacy, and quality of life in older women with OA. CLINICAL RELEVANCE: The sustained effects of acquired exercise behaviors that persisted up to 4 weeks after the program ended could be of interest to rehabilitation nurses and other healthcare professionals.
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Exercícios de Alongamento Muscular/psicologia , Osteoartrite/terapia , Autoeficácia , Idoso , Idoso de 80 Anos ou mais , Índice de Massa Corporal , Depressão/complicações , Depressão/psicologia , Feminino , Humanos , Vida Independente/psicologia , Vida Independente/estatística & dados numéricos , Masculino , Osteoartrite/psicologia , Projetos Piloto , Qualidade de Vida/psicologia , República da Coreia , Inquéritos e QuestionáriosRESUMO
PURPOSE: This study analyzed the changes in the number of surgeries and surgical patterns due to the adoption and diffusion of new medical technology while focusing on radical prostatectomy. MATERIALS AND METHODS: Medical equipment status report data and the National Health Insurance claims data from 2007 to 2019 were used. A total of 62798 radical prostatectomies from 135 medical facilities were analyzed. Radical prostatectomy was classified into open radical prostatectomy (ORP), laparoscopic radical prostatectomy (LRP), and robot-assisted laparoscopic radical prostatectomy (RARP) using the fee schedule codes. A linear mixed model was used to determine whether the adoption of a robotic surgical system had an effect on the number of surgeries and surgical patterns after adjusting for medical characteristics. RESULTS: The number of radical prostatectomies performed in Korea increased from 1756 in 2007 to 8475 in 2019. During this period, the proportion of RARP in total surgery increased from 17.5% to 74.3%. The mean number of surgeries at medical facilities adopting the robotic surgical system was 128.3, which was higher compared to 18.5 cases in medical facilities that did not adopt it. The adoption of a robotic surgical system increased the number of radical prostatectomy surgeries by 12.1 cases and the RARP share by 47.2% in a linear mixed model. CONCLUSION: The adoption and diffusion of robotic surgical systems in Korea increased the number of surgeries as well as the share of robotic surgery. It is necessary to manage a technology that is widely used in a state where its clinical effectiveness is uncertain.
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Laparoscopia , Neoplasias da Próstata , Procedimentos Cirúrgicos Robóticos , Humanos , Seguro Saúde , Masculino , Prostatectomia , Neoplasias da Próstata/cirurgia , Resultado do TratamentoRESUMO
MSCs (mesenchymal stem cells) derived from the bone marrow have shown to be a promising source of stem cells in a therapeutic strategy of neurodegenerative disorder. Also, MSCs can enhance the TH (tyrosine hydroxylase) expression and DA (dopamine) content in catecholaminergic cells by in vitro co-culture system. In the present study, we investigated the effect of intrastriatal grafts of MSCs on TH protein and gene levels and DA content in adult intact rats. When MSCs were transplanted into the striatum of normal rats, the grafted striatum not only had significantly higher TH protein and mRNA levels, but also significantly higher DA content than the untransplanted striatum. Meanwhile, the grafted MSCs differentiated into neurons, astrocytes and oligodendrocytes; however, TH-positive cells could not be detected in our study. These experimental results offer further evidence that MSCs are a promising candidate for treating neurodegenerative diseases such as Parkinson's disease.
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Dopamina/metabolismo , Células-Tronco Mesenquimais/metabolismo , Tirosina 3-Mono-Oxigenase/metabolismo , Animais , Células da Medula Óssea/citologia , Diferenciação Celular , Corpo Estriado , Imuno-Histoquímica , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/citologia , RNA Mensageiro/metabolismo , Ratos , Tirosina 3-Mono-Oxigenase/genéticaRESUMO
Development of stable rCHO cell lines is still time consuming and labor intensive, although it is a critical step in the commercial development of recombinant antibodies. The current work demonstrates, for the first time, that electroporation of CHO cells with DMSO can enhance stable expression of recombinant antibodies in rCHO cells. Electroporation with DMSO resulted in an average 3.7-fold and 2.8-fold increases in expression levels of aflibercept and pembrolizumab, respectively, in pools of stable rCHO cells. It also resulted in an average of 2.2-fold and 2.6-fold increases in the expression of aflibercept and pembrolizumab, respectively, in single-cell derived rCHO clones. Simple batch cultures of rCHO cell clones with the highest expression produced 1.0 g/l for aflibercept and 1.4 g/l for pembrolizumab without a time-consuming gene amplification process. Electroporation with DMSO also shortened the development of rCHO cell lines to 2-3 months, allowing rapid establishment of stable rCHO cell lines with a desirable expression level antibodies.
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Anticorpos/metabolismo , Técnicas de Cultura Celular por Lotes/métodos , Dimetil Sulfóxido , Eletroporação , Animais , Anticorpos/genética , Células CHO , Técnicas de Cultura de Células , Cricetulus , Expressão Gênica , Técnicas de Transferência de Genes , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismoRESUMO
Embryonic stem (ES)-like cells were isolated from in vivo-produced cat embryos. Total of 101 blastocysts were collected from female cats. The inner cell mass (ICM) were mechanically isolated and cultured on mitomycin-C-treated cat embryonic fibroblast feeder layers in medium supplemented with knockouttrade mark Serum Replacement (KSR-medium) or fetal bovine serum (FBS-medium). Putative ES-like cell colonies developed in both KSR- and FBS-medium conditions, but formed domed and flat colonies, respectively. ICM cell attachment and ES-like cell colony formation were significantly higher in KSR-medium, but subsequent cell proliferation was significantly lower than in FBS-medium. For passaging, 32 and 18 colonies in KSR- and FBS-medium were separated by enzymatic dissociation or mechanical disaggregation. Enzymatic dissociation resulted in cell differentiation; however, mechanical disaggregation generated cells that remained undifferentiated over more than four passages and yielded two cat ES-like cell lines that continued to grow for up to eight passages in FBS-medium. These cells had typical stem cell morphology, expressed high levels of alkaline phosphatase activity, and were positive for the ES cell-markers Oct-4, stage-specific embryonic antigen-1 (SSEA-1), SSEA-3, and SSEA-4. These cells formed embryoid bodies (EBs) in suspension culture after extended suspension culture. When simple EBs were cultured on tissue culture plates, they differentiated into several cell types, including epithelium-like and neuron-like cells. In addition, EBs were positive for mesoderm marker, desmin. After prolonged in vitro culture, some colonies spontaneously differentiated into beating myocardiocytes, and were positive for alpha-actinin. These observations indicate that cat ES-like cells were successfully isolated and characterized from in vivo-produced blastocysts.
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Blastocisto/citologia , Gatos/fisiologia , Separação Celular , Células-Tronco Embrionárias/citologia , Células-Tronco Embrionárias/fisiologia , Animais , Gatos/embriologia , Técnicas de Cultura de Células , Diferenciação Celular , Separação Celular/métodos , Células Cultivadas , Embrião de Mamíferos , Feminino , Miócitos Cardíacos/fisiologiaRESUMO
Mesenchymal stem cells (MSCs) are pluripotent adult stem cells. It has been shown that MSCs secrete neurotrophic factors involving nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF). Also, these neurotrophic factors can upregulate tyrosine hydroxylase (TH) gene expression in PC12 cells and neural stem cells. Here, we investigated the effect of co-culturing rat E13.5 ventral mesencephalic cells (VMCs) with MSCs from rat bone marrow on TH expression and dopamine (DA) content. The study consisted of 3 groups: MSC, VMC and a combined MSC+VMC group. All groups were cultured in serum-free neuro-basal medium for 3 days. Thereafter, each group was analyzed by RT-PCR, western blotting, and HPLC. The co-culture group showed a higher expression at TH and DA than the VMC group. However, TH and DA were not present in the MSC group. These observations suggest that MSCs could be an alternative source for treating neurodegenerative diseases such as Parkinson's disease (PD).
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Comunicação Celular/fisiologia , Dopamina/metabolismo , Mesencéfalo/metabolismo , Células-Tronco Mesenquimais/metabolismo , Neurônios/metabolismo , Tirosina 3-Mono-Oxigenase/metabolismo , Animais , Células Cultivadas , Técnicas de Cocultura , Masculino , Mesencéfalo/citologia , Transplante de Células-Tronco Mesenquimais/métodos , Fatores de Crescimento Neural/metabolismo , Neurônios/citologia , Doença de Parkinson/cirurgia , Ratos , Ratos Sprague-Dawley , Substância Negra/citologia , Substância Negra/metabolismo , Regulação para Cima/fisiologia , Área Tegmentar Ventral/citologia , Área Tegmentar Ventral/metabolismoRESUMO
Umbilical cord blood (UCB)-derived mesenchymal stem cells (MSCs) are multipotent adult stem cells, which can differentiation into cells of connective tissue and neural lineages. This study investigated the potential for neuronal differentiation of red fluorescent protein (RFP)-transgenic cat UCB-derived MSCs. The cells were cultured in pre-induction medium for 24 hr and in neuronal-induction medium for 72 hr. Immunofluorescent staining showed that 6.85% of the total cells were beta III-tubulin-positive, 3.37% were neurofilament light (NF-L)-positive and 7.04% were neurofilament medium (NF-M)-positive. A beta III-tubulin band was detected by western blot analysis. Our results demonstrate that RFP-transgenic UCB-derived MSCs can be differentiated into neuronal cells in vitro. Thus, RFP-transgenic MSCs could provide alternative tracing material for stem cell transplantation.
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Gatos/sangue , Sangue Fetal/citologia , Células-Tronco Mesenquimais/citologia , Neurônios/citologia , Animais , Animais Geneticamente Modificados , Diferenciação Celular/fisiologia , Citometria de Fluxo/veterinária , Proteínas Luminescentes/química , Proteínas Luminescentes/genética , Microscopia de Fluorescência/veterinária , Proteína Vermelha FluorescenteRESUMO
We recently produced 11 cloned kittens by somatic cell nuclear transfer (SCNT) using fibroblasts from a feline fetus (donor A, three kittens), an adult domestic cat (donor B, one kitten), and a deaf adult Turkish Angora cat (donor C, seven kittens). Two kittens were stillborn and three died a month after birth. The donor C-derived kittens did not share their donor's eye color or deafness. To test whether this and the low cloning success rate are due to epigenetic modifications, we compared the methylation of somatic and placental cells from the cloned cats and domestic normal cats by bisulfite mutagenesis sequencing analysis. The DNA methylation of somatic cells from the cloned kittens ranged from 78.0% to 88.9%, and did not differ significantly depending on whether they were stillborn, died early after birth, or were healthy. Donors B and C showed similar levels of methylation (77.0% and 79.1%, respectively), as did somatic cells from normal domestic and Turkish Angora cats (range, 75.7-88.0%). However, donor A showed less methylation (70.6%) than the somatic cells from the kittens derived from it (range, 82.2-88.9%). Moreover, placental cells from three donor C-derived kittens showed significantly higher DNA methylation (range, 76.7-80.5%) than placental cells from normal domestic cats (range, 64.2-74.9%). Thus, methylation of satellite regions in somatic cells may not be responsible for the stillbirth, early death, or different eye and hearing attributes of cloned cats. However, hypermethylation in the placenta of cloned cats may be responsible for low success rates in cloning cats.