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Mutations in the X-linked cyclin-dependent kinase-like 5 (CDKL5) cause CDKL5 deficiency disorder (CDD), a neurodevelopmental disease characterized by severe infantile seizures and intellectual disability. The absence of CDKL5 in mice causes defective spine maturation that can at least partially explain the cognitive impairment in CDKL5 patients and CDD mouse models. The molecular basis for such defect may depend on the capacity of CDKL5 to regulate microtubule (MT) dynamics through its association with the MT-plus end tracking protein CLIP170 (cytoplasmic linker protein 170). Indeed, we here demonstrate that the absence of CDKL5 causes CLIP170 to be mainly in a closed inactive conformation that impedes its binding to MTs. Previously, the synthetic pregnenolone analogue, pregnenolone-methyl-ether (PME), was found to have a positive effect on CDKL5-related cellular and neuronal defects in vitro. Here, we show that PME induces the open active conformation of CLIP170 and promotes the entry of MTs into dendritic spines in vitro. Furthermore, the administration of PME to symptomatic Cdkl5-knock-out mice improved hippocampal-dependent behavior and restored spine maturation and the localization of MT-related proteins in the synaptic compartment. The positive effect on cognitive deficits persisted for 1 week after treatment withdrawal. Altogether, our results suggest that CDKL5 regulates spine maturation and cognitive processes through its control of CLIP170 and MT dynamics, which may represent a novel target for the development of disease-modifying therapies.
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Síndromes Epilépticos , Proteínas Asociadas a Microtúbulos , Proteínas de Neoplasias , Pregnenolona , Animales , Síndromes Epilépticos/genética , Éteres/metabolismo , Hipocampo/metabolismo , Ratones , Proteínas Asociadas a Microtúbulos/genética , Microtúbulos/metabolismo , Proteínas de Neoplasias/genética , Pregnenolona/farmacología , Proteínas Serina-Treonina Quinasas/genéticaRESUMEN
AIM(S): To discuss the methodological aspects of participatory design, arguing for a three-phase approach and the suitability of situating participatory design within a phenomenological-hermeneutical tradition in health science. DESIGN AND METHODS: Methodological discussion based on participatory design theory, epistemology and research studies. RESULTS: The epistemological and methodological discussions show how the core values and key elements of participatory design align with the phenomenological-hermeneutical approach. In addition, examples of participatory design studies are provided to illustrate how it can be conducted in health science. CONCLUSION: Participatory design is a flexible framework based on genuine participation, defined by three core values: having a say, mutual learning and democratization. The iterative processes allow for adjustments in alignment with the core values and the scientific stance that defines the choice of methods, tools and techniques. A phenomenological-hermeneutic approach in participatory design studies is relevant and aligned with the core values of participatory design. Thus, this paper argues for a close integration between the participatory design methodology and the phenomenological-hermeneutic scientific approach within health science. IMPLICATIONS FOR THE PROFESSION: Participatory design is a powerful methodology with core values that can co-design sustainable health technologies with potential to impact patient care and the clinical practice of nurses. When combined with qualitative research methods, patients' lived experiences serve as the foundation for improving clinical nursing practice. Discussing the epistemological aspects of participatory design provides nurse researchers with a coherent methodological understanding, essential for the continual development of nursing research. IMPACT: This paper discusses the research methodology of participatory design within health sciences. It aims to address the lack of understanding of the methodology, particularly within a specific scientific stance. The main finding is the elaboration on participatory design and the relevance of a phenomenological-hermeneutical approach. The paper has the potential to impact researchers, master's and PhD students, as well as others engaged in participatory design or other methodologies related to user involvement within health science. REPORTING METHOD: No available EQUATOR guidelines were applicable to this methodological paper, as no new data were created or analysed. PATIENT OR PUBLIC CONTRIBUTION: There was no direct patient or public contribution, as this is a methodological paper.
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BACKGROUND: Pressure ulcers (PUs) are frequently reported in people with spinal cord injuries (SCI). Wound management in people with SCI involves relieving pressure on the affected area by means of immobilisation and bed rest. The healing time of a PU can vary, but often takes several months or even years, causing people to stay in bed for prolonged periods of time. OBJECTIVE: This study aims to explore the perspectives and lived experiences of people with SCI who are affected by PUs. DESIGN: and method: This study is a qualitative explorative study that employs individual semi-structured in-depth interviews to obtain the narratives of people with SCI and a pressure ulcer. We used a phenomenological-hermeneutic approach that was inspired by Ricoeur's theory of interpretation. The analysis was performed in three levels: Naïve reading, structural analysis and critical interpretation and discussion. PARTICIPANTS: and setting: Ten people with SCI who were being treated in the Danish healthcare system for their PU participated in this study: six participants had experienced a complete traumatic SCI, three had an incomplete traumatic SCI, and one had a non-traumatic complete SCI. The study included nine men and one woman, aged 49-81 years (mean 64). Nine had a PU in the seating area, while one had the ulcer on the leg. RESULTS: The analysis revealed three themes: 1. Struggling to balance prevention with an active, meaningful life, 2. Challenges and consequences of pressure relief protocols and bed rest, 3. Experiencing prolonged and incoherent treatment with varying levels of staff engagement and competencies. CONCLUSIONS: People with SCI and a PU have difficulty balancing their active, redefined lives when subjected to a strict pressure relief protocol. The consequences of immobility caused by pressure relief include reduced social and community participation and decreased quality of life. PU treatment is experienced as incoherent and unnecessarily lengthy, leading to a deterioration in the wounds. Improving PU treatment for people with SCI is of utmost importance and has the potential to benefit not only the people with SCI but also the healthcare system and the economy.
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Úlcera por Presión , Traumatismos de la Médula Espinal , Masculino , Femenino , Humanos , Úlcera por Presión/etiología , Úlcera por Presión/prevención & control , Calidad de Vida , Traumatismos de la Médula Espinal/complicaciones , Investigación Cualitativa , Supuración/complicacionesRESUMEN
AIM: To explore patients' and healthcare professionals' (HCPs) experiences of oral care during hospitalisation to identify needs and challenges. BACKGROUND: Daily oral care is important to patients' health and well-being, to prevent diseases in the oral cavity, systemic infections and increased morbidity, which subsequently can lead to prolonged hospitalisation and, at worst, increased mortality. Despite this knowledge, oral care is a neglected part of nursing practice. Studies do not clearly identify barriers regarding oral care, as the existing knowledge is inadequate. DESIGN: A qualitative study exploring participants' experiences to gain new in-depth knowledge of oral care among hospitalised patients. METHODS: A phenomenological-hermeneutic approach was applied. Participant observations were conducted on five hospital wards, combined with individual semi-structured interviews with 16 patients and 15 HCP. Data analysis was based on Ricoeur's theory of narrative and interpretation. RESULTS: Four themes describing the challenges regarding oral care emerged: Oral care as a gut feeling; oral care fades into the background; even self-reliant patients need help with oral care; and the mouth reflects the life lived. CONCLUSIONS: The identified challenges show there is a need for improvement in the health professional approach to oral care in nursing practice. Focus on increasing HCPs' knowledge, skills and competences can increase their nursing agency and support patients' self-care capacity. IMPACT: Investigation of oral care during hospitalisation revealed four main challenges concerning both patients' and HCPs' lack of knowledge and awareness of oral care. Thus, patients and HCPs should be included in developing solutions to improve oral care in nursing practice. REPORTING METHODS: The COREQ criteria for reporting qualitative research were adhered to. PATIENT CONTRIBUTION: A patient representative was involved in the discussion of the proposal, conduct and results of the study.
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Emociones , Personal de Salud , Humanos , Investigación Cualitativa , Hermenéutica , Atención a la SaludRESUMEN
An extensive multi-country outbreak of multidrug-resistant monophasic Salmonella Typhimurium infection in 10 countries with 150 reported cases, predominantly affecting young children, has been linked to chocolate products produced by a large multinational company. Extensive withdrawals and recalls of multiple product lines have been undertaken. With Easter approaching, widespread product distribution and the vulnerability of the affected population, early and effective real-time sharing of microbiological and epidemiological information has been of critical importance in effectively managing this serious food-borne incident.
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Chocolate , Salmonella typhimurium , Niño , Preescolar , Brotes de Enfermedades , Humanos , Salmonella typhimurium/genética , Reino Unido/epidemiologíaRESUMEN
CDKL5 deficiency disorder (CDD) is an X-linked neurodevelopmental disorder characterised by early-onset drug-resistant epilepsy and impaired cognitive and motor skills. CDD is caused by mutations in cyclin-dependent kinase-like 5 (CDKL5), which plays a well-known role in regulating excitatory neurotransmission, while its effect on neuronal inhibition has been poorly investigated. We explored the potential role of CDKL5 in the inhibitory compartment in Cdkl5-KO male mice and primary hippocampal neurons and found that CDKL5 interacts with gephyrin and collybistin, two crucial organisers of the inhibitory postsynaptic sites. Through molecular and electrophysiological approaches, we demonstrated that CDKL5 loss causes a reduced number of gephyrin puncta and surface exposed γ2 subunit-containing GABAA receptors, impacting the frequency of miniature inhibitory postsynaptic currents, which we ascribe to a postsynaptic function of CDKL5. In line with previous data showing that CDKL5 loss impacts microtubule (MT) dynamics, we showed that treatment with pregnenolone-methyl-ether (PME), which promotes MT dynamics, rescues the above defects. The impact of CDKL5 deficiency on inhibitory neurotransmission might explain the presence of drug-resistant epilepsy and cognitive defects in CDD patients. Moreover, our results may pave the way for drug-based therapies that could bypass the need for CDKL5 and provide effective therapeutic strategies for CDD patients.
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Neuroesteroides , Espasmos Infantiles , Masculino , Ratones , Animales , Neuroesteroides/uso terapéutico , Pregnenolona/farmacología , Espasmos Infantiles/genética , Éteres , Ratones Noqueados , Proteínas Serina-Treonina Quinasas/genéticaRESUMEN
Mutations in the X-linked cyclin-dependent kinase-like 5 (CDKL5) gene cause a complex neurological disorder, characterized by infantile seizures, impairment of cognitive and motor skills and autistic features. Loss of Cdkl5 in mice affects dendritic spine maturation and dynamics but the underlying molecular mechanisms are still far from fully understood. Here we show that Cdkl5 deficiency in primary hippocampal neurons leads to deranged expression of the alpha-amino-3-hydroxy-5-methyl-4-iso-xazole propionic acid receptors (AMPA-R). In particular, a dramatic reduction of expression of the GluA2 subunit occurs concomitantly with its hyper-phosphorylation on Serine 880 and increased ubiquitination. Consequently, Cdkl5 silencing skews the composition of membrane-inserted AMPA-Rs towards the GluA2-lacking calcium-permeable form. Such derangement is likely to contribute, at least in part, to the altered synaptic functions and cognitive impairment linked to loss of Cdkl5. Importantly, we find that tianeptine, a cognitive enhancer and antidepressant drug, known to recruit and stabilise AMPA-Rs at the synaptic sites, can normalise the expression of membrane inserted AMPA-Rs as well as the number of PSD-95 clusters, suggesting its therapeutic potential for patients with mutations in CDKL5.
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Síndromes Epilépticos/tratamiento farmacológico , Proteínas Serina-Treonina Quinasas/genética , Receptores AMPA/genética , Espasmos Infantiles/tratamiento farmacológico , Tiazepinas/administración & dosificación , Animales , Antidepresivos/administración & dosificación , Homólogo 4 de la Proteína Discs Large/genética , Síndromes Epilépticos/genética , Síndromes Epilépticos/patología , Regulación de la Expresión Génica/efectos de los fármacos , Hipocampo/efectos de los fármacos , Hipocampo/fisiopatología , Humanos , Ratones , Mutación , Neurogénesis/efectos de los fármacos , Neuronas/efectos de los fármacos , Neuronas/patología , Fosforilación , Cultivo Primario de Células , Proteínas Serina-Treonina Quinasas/deficiencia , Espasmos Infantiles/genética , Espasmos Infantiles/patología , Sinapsis/efectos de los fármacos , Sinapsis/genéticaRESUMEN
BACKGROUND: Extrauterine growth restriction (EUGR) in preterm infants is associated with higher morbidity and impaired neurodevelopment. Early nutrition support may prevent EUGR in preterm infants, but it is not known if this improves organ development and brain function in the short and long term. OBJECTIVE: Using pigs as models for infants, we hypothesized that diet-induced EUGR impairs gut, immunity, and brain development in preterm neonates during the first weeks after birth. METHODS: Forty-four preterm caesarean-delivered pigs (Danish Landrace × Large White × Duroc, birth weight 975 ± 235 g, male:female ratio 23:21) from 2 sows were fed increasing volumes [32-180 mL/(kg·d)] of dilute bovine milk (EUGR group) or the same diet fortified with powdered bovine colostrum for 19 d (CONT group, 50-100% higher protein and energy intake than the EUGR group). RESULTS: The EUGR pigs showed reduced body growth (-39%, P < 0.01), lower plasma albumin, phosphate, and creatine kinase concentrations (-35 to 14%, P < 0.05), increased cortisol and free iron concentrations (+130 to 700%, P < 0.05), and reduced relative weights of the intestine, liver, and spleen (-38 to 19%, all P < 0.05). The effects of EUGR on gut structure, function, microbiota, and systemic immunity were marginal, although EUGR temporarily increased type 1 helper T cell (Th1) activity (e.g. more blood T cells and higher Th1-related cytokine concentrations on day 8) and reduced colon nutrient fermentation (lower SCFA concentration; -45%, P < 0.01). Further, EUGR pigs showed increased relative brain weights (+19%, P < 0.01), however, memory and learning, as tested in a spatial T-maze, were not affected. CONCLUSION: Most of the measured organ growth, and digestive, immune, and brain functions showed limited effects of diet-induced EUGR in preterm pigs during the first weeks after birth. Likewise, preterm infants may show remarkable physiological adaptation to deficient nutrient supply during the first weeks of life although early life malnutrition may exert negative consequences later.
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Animales Recién Nacidos/crecimiento & desarrollo , Encéfalo/crecimiento & desarrollo , Tracto Gastrointestinal/crecimiento & desarrollo , Inmunidad/fisiología , Necesidades Nutricionales , Sus scrofa/crecimiento & desarrollo , Animales , Calostro , Femenino , Microbioma Gastrointestinal , Tracto Gastrointestinal/anatomía & histología , Edad Gestacional , Humanos , Fenómenos Fisiológicos Nutricionales del Lactante , Recién Nacido , Recien Nacido Prematuro/crecimiento & desarrollo , Masculino , Leche , Modelos Animales , Apoyo Nutricional , Valor NutritivoRESUMEN
AIM: To explore patients' and healthcare professionals' experiences of using a telehealth solution developed to improve follow-up after kidney transplantation. BACKGROUND: Transplantation is the treatment of choice whenever feasible for patients with end-stage kidney disease. However, it implies lifelong adherence of self-monitoring, medicine and other restrictions to ensure successful outcomes. Based on user involvement, a telehealth solution was developed to support patients and healthcare professionals post-transplantation. DESIGN: An explorative qualitative study with a phenomenological-hermeneutic approach. METHODS: The developed app and workflow for follow-up were tested by patients and healthcare professionals and evaluated with interviews. In total, 16 patients and 20 healthcare professionals participated. Individual interviews were conducted with the patients, four nurses participated in two sets of interviews, and 16 doctors participated in a focus group. Data were analysed with inspiration from Ricoeur's theory of interpretation, on three levels: Naïve reading, structural analysis and critical interpretation and discussion. The COREQ checklist was applied in reporting the study. RESULTS: Three themes emerged: Challenging conditions for training sessions, telehealth improves patient reflection and collaboration, and telehealth gives patients a voice in consultations. In a challenging time, post-transplantation patients found the app easy to use; it facilitated support and reflection on how to manage. It also supported both patients and healthcare professionals at follow-up consultations in terms of enhanced preparation, improved dialogue and enabling consultations by phone. CONCLUSION: The study showed that patients and healthcare professionals found the app and workflow valuable and easy to use. The Patient Data feature in the app has potential as a communication tool. However, adjustments and further investigations are needed to develop the solution. RELEVANCE TO CLINICAL PRACTICE: The potential of telehealth brings new opportunities to provide treatment and care to newly transplanted patients. Telehealth can support both patients and health professionals by improving dialogue and collaboration.
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Trasplante de Riñón/enfermería , Telemedicina/métodos , Receptores de Trasplantes , Adulto , Femenino , Grupos Focales , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Investigación CualitativaRESUMEN
CDKL5 is a protein kinase that plays a key role for neuronal functions as testified by the onset of complex neuronal dysfunctions in patients with genetic lesions in CDKL5. Here we identify a novel interactor of CDKL5, IQGAP1, a fundamental regulator of cell migration and polarity. In accordance with a functional role of this interaction, depletion of CDKL5 impairs cell migration and impedes the localization of IQGAP1 at the leading edge. Moreover, we demonstrate that CDKL5 is required for IQGAP1 to form a functional complex with its effectors, Rac1 and the microtubule plus end tracking protein CLIP170. These defects eventually impact on the microtubule association of CLIP170, thus deranging their dynamics. CLIP170 is a cellular target of the neurosteroid pregnenolone; by blocking CLIP170 in its active conformation, pregnenolone is capable of restoring the microtubule association of CLIP170 in CDKL5 deficient cells and rescuing morphological defects in neurons devoid of CDKL5. These findings provide novel insights into CDKL5 functions and pave the way for target-specific therapeutic strategies for individuals affected with CDKL5-disorder.
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Pregnenolona/metabolismo , Proteínas Serina-Treonina Quinasas/genética , Proteínas Serina-Treonina Quinasas/metabolismo , Actinas/metabolismo , Animales , Células COS , Movimiento Celular/fisiología , Chlorocebus aethiops , Células HeLa , Humanos , Ratones , Proteínas Asociadas a Microtúbulos/metabolismo , Microtúbulos/metabolismo , Microtúbulos/patología , Proteínas de Neoplasias/metabolismo , Neuronas/metabolismo , Neurotransmisores/metabolismo , Unión Proteica , Proteínas Activadoras de ras GTPasa/genética , Proteínas Activadoras de ras GTPasa/metabolismoRESUMEN
AIM: The aim of this study is to compare the effect of exercise training on physical capacity and alcohol consumption in alcohol use disorder (AUD) patients. METHODS: One hundred and five AUD patients were randomly assigned to treatment as usual combined with running and brisk walking for 30-45 min twice a week, either in small supervised groups (GR) or individually (IND), or to a control group with no running (C). Assessments were made after 6 and 12 months of training. RESULTS: Training volume was estimated as 36 min per training bout at an intensity of 78% of HRmax with no differences between GR and IND ( p>.05). A highly significant reduction in training frequency was seen in both training groups after the first month ( p<.0001). Only IND increased VO2max, by 5.7% ( p<.05), while no differences were seen between GR, IND and C. Alcohol intake decreased from 219 to 41 units per 30 days as the average for the entire sample with no significant difference of drinking outcomes between groups ( p<.0001). CONCLUSIONS: We saw an effect on drinking habits after running in both groups. However, no additional effect was seen when compared with the control group. A drop in the training frequency during the intervention might have resulted in an insignificant training stimulus.
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Consumo de Bebidas Alcohólicas/psicología , Alcoholismo/terapia , Ejercicio Físico/psicología , Adulto , Femenino , Humanos , Masculino , Persona de Mediana Edad , Resultado del TratamientoRESUMEN
CDKL5 deficiency disorder (CDD) is a severe neurodevelopmental encephalopathy caused by mutations in the X-linked CDKL5 gene that encodes a serine/threonine kinase. CDD is characterised by the early onset of seizures and impaired cognitive and motor skills. Loss of CDKL5 in vitro and in vivo affects neuronal morphology at early and late stages of maturation, suggesting a link between CDKL5 and the neuronal cytoskeleton. Recently, various microtubule (MT)-binding proteins have been identified as interactors of CDKL5, indicating that its roles converge on regulating MT functioning. MTs are dynamic structures that are important for neuronal morphology, migration and polarity. The delicate control of MT dynamics is fundamental for proper neuronal functions, as evidenced by the fact that aberrant MT dynamics are involved in various neurological disorders. In this review, we highlight the link between CDKL5 and MTs, discussing how CDKL5 deficiency may lead to deranged neuronal functions through aberrant MT dynamics. Finally, we discuss whether the regulation of MT dynamics through microtubule-targeting agents may represent a novel strategy for future pharmacological approaches in the CDD field.
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Síndromes Epilépticos/metabolismo , Microtúbulos/metabolismo , Neuronas/metabolismo , Espasmos Infantiles/metabolismo , Animales , Humanos , Microtúbulos/efectos de los fármacos , Neuronas/efectos de los fármacos , Pregnenolona/farmacologíaRESUMEN
Preterm birth interrupts intrauterine brain growth and maturation and may induce a delay in postnatal neurodevelopment. Such developmental delays can result from the reduced fetal age at birth, together with the clinical compli-cations of preterm birth (e.g., hypoxia, ischemia, and inflammation). We hypothesized that late preterm birth, inducing only mild clinical complications, has minimal effects on brain-related outcomes such as motor function and behavior. Using the pig as a model for late preterm infants, piglets were cesarean delivered preterm (90%, 106 days gestation) or at full term, reared by identical procedures, and euthanized for tissue collection at birth or after 11 days (e.g., term-corrected age for preterm pigs). Clinical variables and both structural and functional brain endpoints were assessed. The preterm pigs were slow to get on their feet, gained less weight (-30%), and had a higher cerebral hydration level and blood-to-cerebrospinal fluid permeability than the term pigs. At term-corrected age (11 days), the absolute weight of the brain and the weights of its regions were similar between 11-day-old preterm and newborn term pigs, and both were lower than in 11-day-old term pigs. Postnatally, physical activity and movements in an open field were similar, except that preterm pigs showed a reduced normalized stride length and increased normalized maximum stride height. Perinatal brain growth is closely associated with advancing postconceptional age in pigs, and late preterm birth is initially associated with impaired brain growth and physical activity. Postnatally, neuromuscular functions mature rapidly and become similar to those in term pigs, even before term-corrected age. Neuromuscular functions and behavior may show rapid postnatal adaptation to late preterm birth in both pigs and infants.
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OBJECTIVE: Pericytes surround the endothelial cells of the microvasculature where they serve as active participants in crucial vascular functions such as angiogenesis, stability, and permeability. However, pericyte loss or dysfunction has been described in a number of pathologies. Targeting pericytes could therefore prove instrumental in the further development of vascular therapeutics. METHODS: To target the pericyte, a proteomic-based approach using antibody phage display was conducted. We present a novel single-cell selection strategy, with a modified selection step to drive the selection of antibodies toward relevant pericyte epitopes. RESULTS: Characterization of the selected antibodies revealed two antibodies with binding specificity for pericytes. The cognate antigen of one of the antibodies was identified as pericyte-expressed fibronectin. This antibody was shown to be a potent inhibitor of pericyte migration and to induce a pro-angiogenic response when included in a pericyte-endothelial cell co-culture angiogenesis assay. CONCLUSIONS: The selection method provides an efficient platform for the selection of functional antibodies which target pericytes. We obtain an antibody that interacts with a fibronectin epitope important for pericyte mobility and functionality. Targeting of this epitope in pathologies where pericytes are implicated could potentially be of therapeutic benefit.
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Anticuerpos/farmacología , Pericitos/efectos de los fármacos , Proteómica/métodos , Anticuerpos/uso terapéutico , Movimiento Celular/efectos de los fármacos , Técnicas de Cocultivo , Fibronectinas/inmunología , Humanos , Neovascularización Fisiológica/efectos de los fármacos , Análisis de la Célula IndividualRESUMEN
MeCP2 is associated with several neurological disorders; of which, Rett syndrome undoubtedly represents the most frequent. Its molecular roles, however, are still unclear, and data from animal models often describe adult, symptomatic stages, while MeCP2 functions during embryonic development remain elusive. We describe the pattern and timing of Mecp2 expression in the embryonic neocortex highlighting its low but consistent expression in virtually all cells and show the unexpected occurrence of transcriptional defects in the Mecp2 null samples at a stage largely preceding the onset of overt symptoms. Through the deregulated expression of ionic channels and glutamatergic receptors, the lack of Mecp2 during early neuronal maturation leads to the reduction in the neuronal responsiveness to stimuli. We suggest that such features concur to morphological alterations that begin affecting Mecp2 null neurons around the perinatal age and become evident later in adulthood. We indicate MeCP2 as a key modulator of the transcriptional mechanisms regulating cerebral cortex development. Neurological phenotypes of MECP2 patients could thus be the cumulative result of different adverse events that are already present at stages when no obvious signs of the pathology are evident and are worsened by later impairments affecting the central nervous system during maturation and maintenance of its functionality.
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Corteza Cerebral/embriología , Corteza Cerebral/metabolismo , Proteína 2 de Unión a Metil-CpG/metabolismo , Neuronas/metabolismo , Animales , Calcio/metabolismo , Movimiento Celular/fisiología , Células Cultivadas , Corteza Cerebral/patología , Progresión de la Enfermedad , Estimulación Eléctrica , Técnica del Anticuerpo Fluorescente , Ácido Glutámico/metabolismo , Proteína 2 de Unión a Metil-CpG/genética , Ratones Noqueados , Neuronas/patología , Síndrome de Rett , Factores de Tiempo , Transcripción Genética , Imagen de Colorante Sensible al VoltajeRESUMEN
Mutations in the X-linked CDKL5 (cyclin-dependent kinase-like 5) gene have been associated with several forms of neurodevelopmental disorders, including atypical Rett syndrome, autism spectrum disorders, and early infantile epileptic encephalopathy. Accordingly, loss of CDKL5 in mice results in autistic-like features and impaired neuronal communication. Although the biological functions of CDKL5 remain largely unknown, recent pieces of evidence suggest that CDKL5 is involved in neuronal plasticity. Herein, we show that, at all stages of development, neuronal depolarization induces a rapid increase in CDKL5 levels, mostly mediated by extrasomatic synthesis. In young neurons, this induction is prolonged, whereas in more mature neurons, NMDA receptor stimulation induces a protein phosphatase 1-dependent dephosphorylation of CDKL5 that is mandatory for its proteasome-dependent degradation. As a corollary, neuronal activity leads to a prolonged induction of CDKL5 levels in immature neurons but to a short lasting increase of the kinase in mature neurons. Recent results demonstrate that many genes associated with autism spectrum disorders are crucial components of the activity-dependent signaling networks regulating the composition, shape, and strength of the synapse. Thus, we speculate that CDKL5 deficiency disrupts activity-dependent signaling and the consequent synapse development, maturation, and refinement.
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Neuronas/metabolismo , Complejo de la Endopetidasa Proteasomal/metabolismo , Proteína Fosfatasa 1/metabolismo , Proteínas Serina-Treonina Quinasas/metabolismo , Receptores de N-Metil-D-Aspartato/metabolismo , Sinapsis/fisiología , Animales , Apoptosis , Western Blotting , Células Cultivadas , Corteza Cerebral/citología , Corteza Cerebral/metabolismo , Electrofisiología , Técnica del Anticuerpo Fluorescente , Hipocampo/citología , Hipocampo/metabolismo , Ratones , Neuronas/citología , Fosforilación , Biosíntesis de Proteínas , Proteína Fosfatasa 1/genética , Proteínas Serina-Treonina Quinasas/genética , Proteolisis , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Receptores de N-Metil-D-Aspartato/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transducción de SeñalRESUMEN
Mutations in MECP2 cause a broad spectrum of neuropsychiatric disorders of which Rett syndrome represents the best defined condition. Both neuronal and non-neuronal functions of the methyl-binding protein underlie the related pathologies. Nowadays MeCP2 is recognized as a multifunctional protein that modulates its activity depending on its protein partners and posttranslational modifications. However, we are still missing a comprehensive understanding of all MeCP2 functions and their involvement in the related pathologies. The study of human mutations often offers the possibility of clarifying the functions of a protein. Therefore, we decided to characterize a novel MeCP2 phospho-isoform (Tyr-120) whose relevance was suggested by a Rett syndrome patient carrying a Y120D substitution possibly mimicking a constitutively phosphorylated state. Unexpectedly, we found MeCP2 and its Tyr-120 phospho-isoform enriched at the centrosome both in dividing and postmitotic cells. The molecular and functional connection of MeCP2 to the centrosome was further reinforced through cellular and biochemical approaches. We show that, similar to many centrosomal proteins, MeCP2 deficiency causes aberrant spindle geometry, prolonged mitosis, and defects in microtubule nucleation. Collectively, our data indicate a novel function of MeCP2 that might reconcile previous data regarding the role of MeCP2 in cell growth and cytoskeleton stability and that might be relevant to understand some aspects of MeCP2-related conditions. Furthermore, they link the Tyr-120 residue and its phosphorylation to cell division, prompting future studies on the relevance of Tyr-120 for cortical development.
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Centrosoma/metabolismo , Proteína 2 de Unión a Metil-CpG/metabolismo , Animales , Células COS , Células Cultivadas , Chlorocebus aethiops , Células HEK293 , Células HeLa , Humanos , Proteína 2 de Unión a Metil-CpG/genética , Ratones , Microtúbulos/metabolismo , Mitosis , Mutación Missense , Fosforilación , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Transporte de Proteínas , Síndrome de Rett/genéticaRESUMEN
BACKGROUND: Chlamydia remains prevalent worldwide and is considered a global public health problem. However, testing rates among young sexually active people remain low. Effective clinical management relies on screening asymptomatic patients. However, attending face-to-face consultations of testing for sexually transmitted infections is associated with stigmatization and anxiety. Self-testing technology (STT) allows patients to test themselves for chlamydia and gonorrhea without the presence of health care professionals. This may result in wider access to testing and increase testing uptake. Therefore, the sexual health clinic at Odense University Hospital has designed and developed a technology that allows patients to get tested at the clinic through self-collected sampling without a face-to-face consultation. OBJECTIVE: This study aimed to (1) pilot-test STT used in clinical practice and (2) investigate the experiences of patients who have completed a self-test for chlamydia and gonorrhea. METHODS: The study was conducted as a qualitative study inspired by the methodology of participatory design. Ethnographic methods were applied in the feasibility study and the data analyzed were inspired by the action research spiral in iterative processes using steps, such as plan, act, observe, and reflect. The qualitative evaluation study used semistructured interviews and data were analyzed using a qualitative 3-level analytical model. RESULTS: The findings from the feasibility study, such as lack of signposting and adequate information, led to the final modifications of the self-test technology and made it possible to implement it in clinical practice. The qualitative evaluation study found that self-testing was seen as more appealing than testing at a face-to-face consultation because it was an easy solution that both saved time and allowed for the freedom to plan the visit independently. Security was experienced when the instructions balanced between being detail-oriented while also being simple and illustrative. The anonymity and discretion contributed to preserving privacy and removed the fear of an awkward conversation or being judged by health care professionals thus leading to the reduction of intrusive feelings. CONCLUSIONS: Accessible health care services are crucial in preventing and reducing the impact of sexually transmitted infections and STT may have the potential to increase testing uptake as it takes into account some of the barriers that exist. The pilot test and evaluation have resulted in a fully functioning implementation of STT in clinical practice.
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Objectives: This article describes how a home visit solution was developed in a co-design process between patients in treatment for severe spasticity, their caregivers and hospital nurses. The solution was developed using a participatory design approach and was based on the identified needs of the participants. Methods: We developed a home visit solution through an iterative process and a collective 'reflection-in-action' approach with patients, caregivers and healthcare professionals. Results: The study revealed the complexities of establishing new routines around home visits. The solution included a new workflow for the nurses and a new route and appointment planning tool. Conclusion: Through a participatory design approach, the users developed a home visit solution that minimised disruption to patients' daily lives and facilitated a dialogue between the nurses and the caregivers about the treatment and the patients' spasticity, which helped to adjust the treatment in line with the patient´s needs.
Asunto(s)
Citas y Horarios , Visita Domiciliaria , Espasticidad Muscular , Humanos , Femenino , Masculino , Persona de Mediana Edad , Cuidadores/psicología , AdultoRESUMEN
AIM: This qualitative study explored the experiences of patients, caregivers and healthcare professionals after replacing hospital consultations with a home service solution for patients treated for severe spasticity with intrathecal baclofen. METHOD: Semi-structured interviews were conducted with six patients, six caregivers and six healthcare professionals. The data were analysed using interpretative phenomenological analysis. RESULTS: Four themes were identified: 1) Caregivers' new role and contribution to treatment, 2) Hospital consultations in home settings changed the balance of power, 3) A compassionate relationship, and 4) Contextual care for patients. CONCLUSION: The home-based consultations improved caregivers' understanding of the treatment and helped them to become active spokespersons for the patient by contributing important information about the patient's perspectives and the effects of the treatment. The balance of power shifted towards a more equitable dynamic between patients and nurses during home consultations, facilitated by greater involvement of the patient perspective. The nurses' working conditions changed, but patients and caregivers appreciated the shift in treatment setting.
Collaboration between nurses and caregivers is important to maintain optimal treatment as caregivers are often spokespersons for the patient and can explain patients' preferences for treatmentHome-based treatment clarifies more of the patient's everyday life, which can then be better incorporated into the nurses' decisions and perspectivesHome-based treatment changes the balance of power between patients, caregivers, and nursesWhen hospital nurse consultations is replaced with consultations at home, there are both practical and organisational implications for nurses' working conditions.