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Neuropsychiatric and neurodegenerative disorders are frequently associated with gastrointestinal (GI) co-pathologies. Although the central and enteric nervous systems (CNS and ENS, respectively) have been studied separately, there is increasing interest in factors that may contribute to conditions affecting both systems. There is compelling evidence that serotonin (5-HT) may play an important role in several gut-brain disorders. It is well known that 5-HT is essential for the development and functioning of the CNS. However, most of the body's 5-HT is produced in the GI tract. A deeper understanding of the specific effects of enteric 5-HT on gut-brain disorders may provide the basis for the development of new therapeutic targets. This review summarizes current data focusing on the important role of 5-HT in ENS development and motility, with particular emphasis on novel aspects of 5-HT signaling in conditions where CNS and ENS comorbidities are common, such as Parkinson's disease and depressive disorders.
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Altered development and function of the prefrontal cortex (PFC) during adolescence is implicated in the origin of mental disorders. Deficits in the GABAergic system prominently contribute to these alterations. Nav1.1 is a voltage-gated Na+ channel critical for normal GABAergic activity. Here, we studied the role of Nav1.1 in PFC function and its potential relationship with the aetiology of mental disorders. Dysfunction of Nav1.1 activity in the medial PFC (mPFC) of adolescent mice enhanced the local excitation/inhibition ratio, resulting in epileptic activity, cognitive deficits and depressive-like behaviour in adulthood, along with a gene expression profile linked to major depressive disorder (MDD). Additionally, it reduced extracellular serotonin concentration in the dorsal raphe nucleus and brain-derived neurotrophic factor expression in the hippocampus, two MDD-related brain areas beyond the PFC. We also observed alterations in oscillatory activity and impaired hippocampal-mPFC coherence during sleep. Finally, we found reduced expression levels of SCN1A, the gene encoding Nav1.1, in post-mortem PFC samples from human MDD subjects. Collectively, our results provide a novel mechanistic framework linking adolescence-specific alterations in Nav1.1 function in the PFC to the pathogenesis of epilepsy and comorbidities such as cognitive impairment and depressive disorders.
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Depression is a devastating mood disorder that causes significant disability worldwide. Current knowledge of its pathophysiology remains modest and clear biological markers are lacking. Emerging evidence from human and animal models reveals persistent alterations in endoplasmic reticulum (ER) homeostasis, suggesting that ER stress-related signaling pathways may be targets for prevention and treatment. However, the neurobiological basis linking the pathways involved in depression-related ER stress remains unknown. Here, we report that an induced model of ER stress in mouse serotonin (5-HT) neurons is associated with reduced Egr1-dependent 5-HT cellular activity and 5-HT neurotransmission, resulting in neuroplasticity deficits in forebrain regions and a depressive-like phenotype. Ketamine administration engages downstream eIF2α signaling to trigger rapid neuroplasticity events that rescue the depressive-like effects. Collectively, these data identify ER stress in 5-HT neurons as a cellular pathway involved in the pathophysiology of depression and show that eIF2α is critical in eliciting ketamine's fast antidepressant effects.
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Antipsychotic drugs of different chemical/pharmacological families show preferential dopamine (DA) D2 receptor (D2-R) vs. D1 receptor (D1-R) affinity, with the exception of clozapine, the gold standard of schizophrenia treatment, which shows a comparable affinity for both DA receptors. Here, we examined the ability of Lu AF35700 (preferential D1-R>D2-R antagonist), to reverse the alterations in thalamo-cortical activity induced by phencyclidine (PCP), used as a pharmacological model of schizophrenia. Lu AF35700 reversed the PCP-induced alteration of neuronal discharge and low frequency oscillation (LFO, 0.15-4 Hz) in thalamo-cortical networks. Likewise, Lu AF35700 prevented the increased c-fos mRNA expression induced by PCP in thalamo-cortical regions of awake rats. We next examined the contribution of D1-R and D2-R to the antipsychotic reversal of PCP effects. The D2-R antagonist haloperidol reversed PCP effects on thalamic discharge rate and LFO. Remarkably, the combination of sub-effective doses of haloperidol and SCH-23390 (DA D1-R antagonist) fully reversed the PCP-induced fall in thalamo-cortical LFO. However, unlike with haloperidol, SCH-23390 elicited different degrees of potentiation of the effects of low clozapine and Lu AF35700 doses. Overall, the present data support a synergistic interaction between both DA receptors to reverse the PCP-induced alterations of oscillatory activity in thalamo-cortical networks, possibly due to their simultaneous blockade in direct and indirect pathways of basal ganglia. The mild potentiation induced by SCH-23390 in the case of clozapine and Lu AF35700 suggests that, at effective doses, these agents reverse PCP effects through the simultaneous blockade of both DA receptors.
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Antipsicóticos , Clozapina , Ratas , Animales , Fenciclidina/farmacología , Clozapina/farmacología , Haloperidol/farmacología , Dopamina , Antipsicóticos/farmacología , Antagonistas de Dopamina/farmacología , Receptores de Dopamina D1RESUMEN
The reciprocal connectivity between the medial prefrontal cortex (mPFC) and the dorsal raphe nucleus (DR) is involved in mood control and resilience to stress. The infralimbic subdivision (IL) of the mPFC is the rodent equivalent of the ventral anterior cingulate cortex, which is intimately related to the pathophysiology/treatment of major depressive disorder (MDD). Boosting excitatory neurotransmission in the IL-but not in the prelimbic cortex, PrL-evokes depressive-like or antidepressant-like behaviors in rodents, which are associated with changes in serotonergic (5-HT) neurotransmission. We therefore examined the control of 5-HT activity by both of the mPFC subdivisions in anesthetized rats. The electrical stimulation of IL and PrL at 0.9 Hz comparably inhibited 5-HT neurons (53% vs. 48%, respectively). However, stimulation at higher frequencies (10-20 Hz) revealed a greater proportion of 5-HT neurons sensitive to IL than to PrL stimulation (86% vs. 59%, at 20 Hz, respectively), together with a differential involvement of GABAA (but not 5-HT1A) receptors. Likewise, electrical and optogenetic stimulation of IL and PrL enhanced 5-HT release in DR in a frequency-dependent manner, with greater elevations after IL stimulation at 20 Hz. Hence, IL and PrL differentially control serotonergic activity, with an apparent superior role of IL, an observation that may help to clarify the brain circuits involved in MDD.
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Trastorno Depresivo Mayor , Núcleo Dorsal del Rafe , Ratas , Animales , Corteza Cerebral , Neuronas , Corteza Prefrontal/fisiologíaRESUMEN
A colloidal synthesis' proof-of-concept based on the Bligh-Dyer emulsion inversion method was designed for integrating into lipid nanoparticles (LNPs) cell-permeating DNA antisense oligonucleotides (ASOs), also known as GapmeRs (GRs), for mRNA interference. The GR@LNPs were formulated to target brain border-associated macrophages (BAMs) as a central nervous system (CNS) therapy platform for silencing neuroinflammation-related genes. We specifically aim at inhibiting the expression of the gene encoding for lipocalin-type prostaglandin D synthase (L-PGDS), an anti-inflammatory enzyme expressed in BAMs, whose level of expression is altered in neuropsychopathologies such as depression and schizophrenia. The GR@LNPs are expected to demonstrate a bio-orthogonal genetic activity reacting with L-PGDS gene transcripts inside the living system without interfering with other genetic or biochemical circuitries. To facilitate selective BAM phagocytosis and avoid subsidiary absorption by other cells, they were functionalized with a mannosylated lipid as a specific MAN ligand for the mannose receptor presented by the macrophage surface. The GR@LNPs showed a high GR-packing density in a compact multilamellar configuration as structurally characterized by light scattering, zeta potential, and transmission electronic microscopy. As a preliminary biological evaluation of the mannosylated GR@LNP nanovectors into specifically targeted BAMs, we detected in vivo gene interference after brain delivery by intracerebroventricular injection (ICV) in Wistar rats subjected to gene therapy protocol. The results pave the way towards novel gene therapy platforms for advanced treatment of neuroinflammation-related pathologies with ASO@LNP nanovectors.
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Stopping the spread of coronavirus disease 2019 (COVID-19) is critical and can be achieved through rapid and effective detection techniques. Our objective was to compare the diagnostic accuracy of rapid antigen tests (RAgT) and reverse transcriptionquantitative polymerase chain reaction (RT-qPCR) and to describe amplification cycle thresholds (Cts). Participants were children aged 1 month to 11 years with symptoms for less than 7 days, who did not have a detectable result in the past 90 days, and were immunocompetent. A total of 1855 patients were included; the prevalence of COVID-19 was 4.7%. For the RAgT, overall sensitivity was 60.2% and specificity, 99.8%; in children older than 5 years, values were 69.8% and 99.8%, respectively. Ct values for discordant samples were higher. To conclude, the diagnostic accuracy indicated that the specificity of RAgT is similar to that of RT-qPCR, but its sensitivity is notably lower, especially in children younger than 5 years.
Frenar la propagación de la enfermedad por el coronavirus 2019 (COVID-19, por su sigla en inglés) es fundamental, y se puede realizar mediante técnicas de detección rápidas y efectivas. El objetivo fue comparar la precisión diagnóstica de un test rápido de antígeno (TRAg,) con la reacción en cadena de polimerasa con retrotranscripción (RT-qPCR, por su sigla en inglés) y describir los umbrales de amplificación (Ct, por su sigla en inglés). Participaron niños de 1 mes a 11 años que tuvieran menos de 7 días de síntomas, sin resultado detectable en los últimos 90 días, e inmunocompetentes. Se incluyeron 1855 pacientes con una prevalencia de COVID-19 del 4,7 %. La sensibilidad global del TRAg fue del 60,2 % y su especificidad, del 99,8 %; en niños mayores de 5 años los valores fueron de 69,8 % y 99,8 %, respectivamente. Los valores de Ct de las muestras discordantes fueron más altos. En conclusión, la precisión diagnóstica muestra que TRAg tiene una especificidad similar a la RT-qPCR, pero una sensibilidad considerablemente menor, sobre todo en niños de menos de 5 años.
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COVID-19 , SARS-CoV-2 , COVID-19/diagnóstico , Prueba de COVID-19 , Niño , Preescolar , Técnicas de Laboratorio Clínico/métodos , Humanos , Reacción en Cadena en Tiempo Real de la Polimerasa , Sensibilidad y EspecificidadRESUMEN
Frenar la propagación de la enfermedad por el coronavirus 2019 (COVID-19, por su sigla en inglés) es fundamental, y se puede realizar mediante técnicas de detección rápidas y efectivas. El objetivo fue comparar la precisión diagnóstica de un test rápido de antígeno (TRAg,) con la reacción en cadena de polimerasa con retrotranscripción (RT-qPCR, por su sigla en inglés) y describir los umbrales de amplificación (Ct, por su sigla en inglés). Participaron niños de 1 mes a 11 años que tuvieran menos de 7 días de síntomas, sin resultado detectable en los últimos 90 días, e inmunocompetentes. Se incluyeron 1855 pacientes con una prevalencia de COVID-19 del 4,7 %. La sensibilidad global del TRAg fue del 60,2 % y su especificidad, del 99,8 %; en niños mayores de 5 años los valores fueron de 69,8 % y 99,8 %, respectivamente. Los valores de Ct de las muestras discordantes fueron más altos. En conclusión, la precisión diagnóstica muestra que TRAg tiene una especificidad similar a la RT-qPCR, pero una sensibilidad considerablemente menor, sobre todo en niños de menos de 5 años.
Stopping the spread of coronavirus disease 2019 (COVID-19) is critical and can be achieved through rapid and effective detection techniques. Our objective was to compare the diagnostic accuracy of rapid antigen tests (RAgT) and reverse transcription quantitative polymerase chain reaction (RT-qPCR) and to describe amplification cycle thresholds (Cts). Participants were children aged 1 month to 11 years with symptoms for less than 7 days, who did not have a detectable result in the past 90 days, and were immunocompetent. A total of 1855 patients were included; the prevalence of COVID-19 was 4.7%. For the RAgT, overall sensitivity was 60.2% and specificity, 99.8%; in children older than 5 years, values were 69.8% and 99.8%, respectively. Ct values for discordant samples were higher. To conclude, the diagnostic accuracy indicated that the specificity of RAgT is similar to that of RT-qPCR, but its sensitivity is notably lower,especially in children younger than 5 years.
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Humanos , Lactante , Preescolar , Niño , SARS-CoV-2 , COVID-19/diagnóstico , Estudios Transversales , Sensibilidad y Especificidad , Técnicas de Laboratorio Clínico/métodos , Reacción en Cadena en Tiempo Real de la Polimerasa , Prueba de COVID-19RESUMEN
Remote detection and monitoring of the vegetation responses to stress became relevant for sustainable agriculture. Ongoing developments in optical remote sensing technologies have provided tools to increase our understanding of stress-related physiological processes. Therefore, this study aimed to provide an overview of the main spectral technologies and retrieval approaches for detecting crop stress in agriculture. Firstly, we present integrated views on: i) biotic and abiotic stress factors, the phases of stress, and respective plant responses, and ii) the affected traits, appropriate spectral domains and corresponding methods for measuring traits remotely. Secondly, representative results of a systematic literature analysis are highlighted, identifying the current status and possible future trends in stress detection and monitoring. Distinct plant responses occurring under shortterm, medium-term or severe chronic stress exposure can be captured with remote sensing due to specific light interaction processes, such as absorption and scattering manifested in the reflected radiance, i.e. visible (VIS), near infrared (NIR), shortwave infrared, and emitted radiance, i.e. solar-induced fluorescence and thermal infrared (TIR). From the analysis of 96 research papers, the following trends can be observed: increasing usage of satellite and unmanned aerial vehicle data in parallel with a shift in methods from simpler parametric approaches towards more advanced physically-based and hybrid models. Most study designs were largely driven by sensor availability and practical economic reasons, leading to the common usage of VIS-NIR-TIR sensor combinations. The majority of reviewed studies compared stress proxies calculated from single-source sensor domains rather than using data in a synergistic way. We identified new ways forward as guidance for improved synergistic usage of spectral domains for stress detection: (1) combined acquisition of data from multiple sensors for analysing multiple stress responses simultaneously (holistic view); (2) simultaneous retrieval of plant traits combining multi-domain radiative transfer models and machine learning methods; (3) assimilation of estimated plant traits from distinct spectral domains into integrated crop growth models. As a future outlook, we recommend combining multiple remote sensing data streams into crop model assimilation schemes to build up Digital Twins of agroecosystems, which may provide the most efficient way to detect the diversity of environmental and biotic stresses and thus enable respective management decisions.
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OBJECTIVE: Recently, we reported on a new MDD-like mouse model based on a regionally selective knockdown of astroglial glutamate transporters, GLAST/GLT-1, in infralimbic cortex (IL) which evokes widespread changes in mouse brain associated with the typical alterations found in MDD patients. To further characterize this new MDD-like mouse model, here we examine some transcriptional elements of glutamatergic/GABAergic neurotransmission and neuroplasticity in forebrain regions in the GLT-1 knockdown mice. Furthermore, we assess the acute ketamine effects on these transcriptional processes. MATERIAL AND METHODS: We used a small interfering RNA (siRNA) pool targeting GLT-1 mRNA to disrupt the GLT-1 transcription in mouse IL. Histological assays were performed to examine postsynaptic density protein-95 (PSD95), neuritin (NRN), glutamine acid descarboxilase-65 (GAD65), and GLT-1 mRNA expression in IL and hippocampus. RESULTS: Knockdown of GLT-1 in mouse IL leads to decreased expression of PSD95 and NRN neuroplasticity mRNAs in IL and hippocampus, which was reversed by an acute dose of ketamine antidepressant. Likewise, a single dose of ketamine also increased the mRNA levels of GAD65 and GLT-1 in IL of GLT-1 knockdown mice, reaching the basal values of control mice. CONCLUSIONS: The glutamatergic neuronal hyperactivity and deficits in the GABA system resulting from siRNA-induced astroglial glutamate transporter knockdown in IL can compromise the integrity/plasticity of neurocircuits affected in MDD. Suitable depressive-like animal models to address the neurobiological changes in MDD are an unmet need and the development of the GLAST/GLT-1 knockdown mouse model may represent a better option to understand the rapid-acting antidepressant effects of ketamine.
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Astrocitos , Ketamina , Plasticidad Neuronal , Sistema de Transporte de Aminoácidos X-AG/metabolismo , Animales , Antidepresivos/farmacología , Antidepresivos/uso terapéutico , Astrocitos/metabolismo , Depresión/genética , Depresión/metabolismo , Transportador 2 de Aminoácidos Excitadores/efectos de los fármacos , Transportador 2 de Aminoácidos Excitadores/genética , Humanos , Ketamina/metabolismo , Ketamina/farmacología , Ketamina/uso terapéutico , Ratones , Plasticidad Neuronal/efectos de los fármacos , Plasticidad Neuronal/genética , ARN Mensajero/metabolismo , ARN Interferente Pequeño/metabolismoRESUMEN
Abstract Human parechovirus (HPeV) is one of the members of the family Picornaviridae that has been associated with fever of unknown origin, gastroenteritis, clinical sepsis, meningitis, orencephalitis in very young infants. HPeV detection is not routinely performed in most clinical microbiology laboratories in Argentina and, therefore, its real prevalence is unknown. We here report three cases of HPeV CNS infection that presented to our hospital with different clinical features after the implementation of a multiplex PCR meningitis/encephalitis panel. Molecular diagnostic techniques could help improve patient care and understand the real prevalence of this infection in Argentina.
Resumen Los parechovirus humanos (HPeV) son virus de la familia Picornaviridae, que se han asociado a diferentes cuadros clínicos, como fiebre de origen desconocido, gastroenteritis, sepsis, meningitis o encefalitis en ninos pequeños. Su detección no está disponible de rutina en la mayoría de los laboratorios de nuestro país, por lo que su prevalencia es desconocida. Reportamos 3 casos de infección del sistema nervioso central por HPeV con diferentes características clínicas, que se presentaron luego de la implementación de un panel molecular para el diagnóstico sindrómico de meningitis/encefalitis. Las técnicas de diagnóstico molecular podrían ayudar a mejorar el abordaje y el cuidado de estos pacientes, así como también a conocer la prevalencia de esta infección en Argentina.
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Anxiety and depression affect 35-50% of patients with Parkinson's disease (PD), often precede the onset of motor symptoms, and have a negative impact on their quality of life. Dysfunction of the serotonergic (5-HT) system, which regulates mood and emotional pathways, occurs during the premotor phase of PD and contributes to a variety of non-motor symptoms. Furthermore, α-synuclein (α-Syn) aggregates were identified in raphe nuclei in the early stages of the disease. However, there are very few animal models of PD-related neuropsychiatric disorders. Here, we develop a new mouse model of α-synucleinopathy in the 5-HT system that mimics prominent histopathological and neuropsychiatric features of human PD. We showed that adeno-associated virus (AAV5)-induced overexpression of wild-type human α-Syn (h-α-Syn) in raphe 5-HT neurons triggers progressive accumulation, phosphorylation, and aggregation of h-α-Syn protein in the 5-HT system. Specifically, AAV5-injected mice displayed axonal impairment in the output brain regions of raphe neurons, and deficits in brain-derived neurotrophic factor (BDNF) expression and 5-HT neurotransmission, resulting in a depressive-like phenotype. Intracerebroventricular treatment with an indatraline-conjugated antisense oligonucleotide (IND-ASO) for four weeks induced an effective and safe reduction of h-α-Syn synthesis in 5-HT neurons and its accumulation in the forebrain, alleviating early deficits of 5-HT function and improving the behavioural phenotype. Altogether, our findings show that α-synucleinopathy in 5-HT neurons negatively affects brain circuits that control mood and emotions, resembling the expression of neuropsychiatric symptoms occurring at the onset of PD. Early preservation of 5-HT function by reducing α-Syn synthesis/accumulation may alleviate PD-related depressive symptoms.
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Serotonina , alfa-Sinucleína , Animales , Modelos Animales de Enfermedad , Humanos , Ratones , Neuronas/metabolismo , Oligonucleótidos/metabolismo , Oligonucleótidos/farmacología , Fenotipo , Prosencéfalo/metabolismo , Calidad de Vida , Serotonina/metabolismo , alfa-Sinucleína/genética , alfa-Sinucleína/metabolismo , alfa-Sinucleína/farmacologíaRESUMEN
The synuclein family consists of α-, ß-, and γ-Synuclein (α-Syn, ß-Syn, and γ-Syn) expressed in the neurons and concentrated in synaptic terminals. While α-Syn is at the center of interest due to its implication in the pathogenesis of Parkinson's disease (PD) and other synucleinopathies, limited information exists on the other members. The current study aimed at investigating the biological role of γ-Syn controlling the midbrain dopamine (DA) function. We generated two different mouse models with: (i) γ-Syn overexpression induced by an adeno-associated viral vector and (ii) γ-Syn knockdown induced by a ligand-conjugated antisense oligonucleotide, in order to modify the endogenous γ-Syn transcription levels in midbrain DA neurons. The progressive overexpression of γ-Syn decreased DA neurotransmission in the nigrostriatal and mesocortical pathways. In parallel, mice evoked motor deficits in the rotarod and impaired cognitive performance as assessed by novel object recognition, passive avoidance, and Morris water maze tests. Conversely, acute γ-Syn knockdown selectively in DA neurons facilitated forebrain DA neurotransmission. Importantly, modifications in γ-Syn expression did not induce the loss of DA neurons or changes in α-Syn expression. Collectively, our data strongly suggest that DA release/re-uptake processes in the nigrostriatal and mesocortical pathways are partially dependent on substantia nigra pars compacta /ventral tegmental area (SNc/VTA) γ-Syn transcription levels, and are linked to modulation of DA transporter function, similar to α-Syn.
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Dopamina , Neuronas Dopaminérgicas , gamma-Sinucleína , Animales , Dopamina/metabolismo , Neuronas Dopaminérgicas/metabolismo , Ratones , Sustancia Negra/metabolismo , Transmisión Sináptica/genética , alfa-Sinucleína/genética , alfa-Sinucleína/metabolismo , gamma-Sinucleína/genética , gamma-Sinucleína/metabolismoRESUMEN
Human parechovirus (HPeV) is one of the members of the family Picornaviridae that has been associated with fever of unknown origin, gastroenteritis, clinical sepsis, meningitis, or encephalitis in very young infants. HPeV detection is not routinely performed in most clinical microbiology laboratories in Argentina and, therefore, its real prevalence is unknown. We here report three cases of HPeV CNS infection that presented to our hospital with different clinical features after the implementation of a multiplex PCR meningitis/encephalitis panel. Molecular diagnostic techniques could help improve patient care and understand the real prevalence of this infection in Argentina.
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Parechovirus , Infecciones por Picornaviridae , Sepsis , Argentina , Niño , Humanos , Lactante , Técnicas de Diagnóstico Molecular , Parechovirus/genética , Infecciones por Picornaviridae/diagnóstico , Infecciones por Picornaviridae/epidemiología , Sepsis/diagnóstico , Sepsis/epidemiologíaRESUMEN
Fast and sustained antidepressant effects of ketamine identified the mammalian target of rapamycin (mTOR) signaling pathway as the main modulator of its antidepressive effects. Thus, mTOR signaling has become integral for the preclinical evaluation of novel compounds to treat depression. However, causality between mTOR and depression has yet to be determined. To address this, we knocked down mTOR expression in mice using an acute intracerebral infusion of small interfering RNAs (siRNA) in the infralimbic (IL) or prelimbic (PrL) cortices of the medial prefrontal cortex (mPFC), and evaluated depressive- and anxious-like behaviors. mTOR knockdown in IL, but not PrL, cortex produced a robust depressive-like phenotype in mice, as assessed in the forced swimming test (FST) and the tail suspension test (TST). This phenotype was associated with significant reductions of mTOR mRNA and protein levels 48 h post-infusion. In parallel, decreased brain-derived neurotrophic factor (BDNF) expression was found bilaterally in both IL and PrL cortices along with a dysregulation of serotonin (5-HT) and glutamate (Glu) release in the dorsal raphe nucleus (DRN). Overall, our results demonstrate causality between mTOR expression in the IL cortex and depressive-like behaviors, but not in anxiety.
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Depresión/psicología , Corteza Prefrontal/metabolismo , Serina-Treonina Quinasas TOR/genética , Serina-Treonina Quinasas TOR/metabolismo , Animales , Factor Neurotrófico Derivado del Encéfalo/genética , Factor Neurotrófico Derivado del Encéfalo/metabolismo , Depresión/genética , Depresión/metabolismo , Modelos Animales de Enfermedad , Núcleo Dorsal del Rafe/metabolismo , Técnicas de Silenciamiento del Gen , Ácido Glutámico/metabolismo , Suspensión Trasera , Masculino , Ratones , Serotonina/metabolismo , NataciónRESUMEN
Resumen Las infecciones de las vías respiratorias inferiores se encuentran entre aquellas en las que el uso inadecuado de antimicrobianos es frecuente, por lo que es fundamental contar con una prueba diagnóstica rápida, sensible y específica. El sistema de FilmArray es un análisis de PCR múltiple con un panel de neumonía que incluye 26 microorganismos y 7 marcadores de resistencia antimicrobiana. Los objetivos de este estudio fueron: a) establecer la correlación entre los cultivos cuantitativos para agentes bacterianos de muestras de vías respiratorias inferiores (MRVB) y la detección fenotípica de mecanismos de resistencia con los correspondientes resultados de FilmArray; b) determinar el cambio terapéutico generado con el informe del resultado inmediato. Se incluyó un total de 194 MRVB correspondientes a 191 pacientes con neumonía y se documentaron 277 bacterias. FilmArray identificó 253/277 (91%) bacterias y 161/277 (58%) se aislaron del cultivo, 58 (23%) coincidieron con el mismo recuento, 116 (46,7%) dieron mayores recuentos con FilmArray y 72 (28,9%) fueron detectadas por este método pero el cultivo fue negativo. Se detectaron marcadores de resistencia antimicrobiana en 63 aislados, pero solo 28 fueron confirmados por métodos fenotípicos. Estos resultados podrían haber provocado cambios en el tratamiento antibiótico en el 74,6% (174/194). FilmArray es una herramienta útil para optimizar el tratamiento antimicrobiano en pacientes con neumonía.
Abstract Lower respiratory tract infections are among those in which the inappropriate use of antimicrobials is common, so it is essential to have a rapid, sensitive and specific diagnostic test. The FilmArray system is a multiplex PCR assay with a pneumonia panel that includes 26 microorganisms and 7 antibiotic resistance markers. The objectives of this study were: a) to establish the correlation between quantitative cultures for bacterial agents from lower respiratory tract samples (MRVB) and the phenotypic detection of resistance mechanisms with the corresponding results of FilmArray b) to determine the therapeutic change generated with the immediate result report. A total of 194 MRVB corresponding to 191 patients with pneumonia were included and 277 bacterial strains were documented. FilmArray identified 253/277 (91%) bacteria and 161/277 (58%) were isolated from culture, 58 (23%) matched the same count, 116 (46.7%) yielded higher counts with FilmArray, and 72 (28.9%) with negative culture were detected by this method. Antibiotic resistance markers were detected in 63 strains, but only 28 were confirmed by phenotypic methods. These results may cause changes in the antimicrobial treatment in 74.6% (174/194). FilmArray is a useful tool to optimize antimicrobial therapy in patients with pneumonia.
Resumo As infecções do trato respiratório inferior estão entre aquelas em que o uso inadequado de antimicrobianos é comum, por isso é essencial um teste diagnóstico rápido, sensível e específico. O sistema FilmArray é um ensaio de PCR multiplo com um painel de pneumonia que inclui 26 microrganismos e 7 marcadores de resistência antimicrobiana. Os objetivos deste estudo foram: a) estabelecer a correlação entre as culturas quantitativas de agentes bacterianos de amostras do trato respiratório inferior (MRVB) e a detecção fenotípica de mecanismos de resistência com os resultados correspondentes do FilmArray b) determinar a alteração terapêutica gerada com o relatório de resultado imediato. Um total de 194 MRVB correspondendo a 191 pacientes com pneumonia foram incluídos e 277 cepas bacterianas foram documentadas. FilmArray identificou 253/277 (91%) bactérias e 161/277 (58%) foram isoladas da cultura, 58 (23%) coincidiram com mesma contagem, 116 (46,7%) deram contagens mais altas com FilmArray e 72 (28,9%) foram detectados por este método, mas a cultura foi negativa. Marcadores de resistência antimicrobiana foram detectados em 63 cepas, mas apenas 28 foram confirmados por métodos fenotípicos. Esses resultados puderam causar alterações no tratamento antibiótico em 74,6% (174/194). FilmArray é uma ferramenta útil para otimizar a terapia antimicrobiana em pacientes com pneumonia..
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Neumonía/diagnóstico , Infecciones/diagnóstico , Antiinfecciosos/administración & dosificación , Resistencia de las Vías RespiratoriasRESUMEN
α-Synuclein (α-Syn) protein is involved in the pathogenesis of Parkinson's disease (PD). Point mutations and multiplications of the α-Syn, which encodes the SNCA gene, are correlated with early-onset PD, therefore the reduction in a-Syn synthesis could be a potential therapy for PD if delivered to the key affected neurons. Several experimental strategies for PD have been developed in recent years using oligonucleotide therapeutics. However, some of them have failed or even caused neuronal toxicity. One limiting step in the success of oligonucleotide-based therapeutics is their delivery to the brain compartment, and once there, to selected neuronal populations. Previously, we developed an indatraline-conjugated antisense oligonucleotide (IND-1233-ASO), that selectively reduces α-Syn synthesis in midbrain monoamine neurons of mice, and nonhuman primates. Here, we extended these observations using a transgenic male mouse strain carrying both A30P and A53T mutant human α-Syn (A30P*A53T*α-Syn). We found that A30P*A53T*α-Syn mice at 4-5 months of age showed 3.5-fold increases in human α-Syn expression in dopamine (DA) and norepinephrine (NE) neurons of the substantia nigra pars compacta (SNc) and locus coeruleus (LC), respectively, compared with mouse α-Syn levels. In parallel, transgenic mice exhibited altered nigrostriatal DA neurotransmission, motor alterations, and an anxiety-like phenotype. Intracerebroventricular IND-1233-ASO administration (100 µg/day, 28 days) prevented the α-Syn synthesis and accumulation in the SNc and LC, and recovered DA neurotransmission, although it did not reverse the behavioral phenotype. Therefore, the present therapeutic strategy based on a conjugated ASO could be used for the selective inhibition of α-Syn expression in PD-vulnerable monoamine neurons, showing the benefit of the optimization of ASO molecules as a disease modifying therapy for PD and related α-synucleinopathies.