Disruption of Interleukin-1β Autocrine Signaling Rescues Complex I Activity and Improves ROS Levels in Immortalized Epithelial Cells with Impaired Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Function

Patients with cystic fibrosis (CF) have elevated concentration of cytokines in sputum and a general inflammatory condition. In addition, CF cells in culture produce diverse cytokines in excess, including IL-1B. We have previously shown that IL-1B, at low doses (~30 pM), can stimulate the expression of CFTR in T84 colon carcinoma cells, through NF-KB signaling. However, at higher doses (>2.5 ng/ml, ~150 pM), IL-1B inhibit CFTR mRNA expression. On the other hand, by using differential display, we found two genes with reduced expression in CF cells, corresponding to the mitochondrial proteins CISD1 and MTND4. The last is a key subunit for the activity of mitochondrial Complex I (mCx-I); accordingly, we later found a reduced mCx-I activity in CF cells. Here we found that IB3-1 cells (CF cells), cultured in serum-free media, secrete 323±5 pg/ml of IL-1B in 24 h vs 127±3 pg/ml for S9 cells (CFTR-corrected IB3-1 cells). Externally added IL-1B (5 ng/ml) reduces the mCx-I activity and increases the mitochondrial (MitoSOX probe) and cellular (DCFH-DA probe) ROS levels of S9 (CFTR-corrected IB3-1 CF cells) or Caco-2/pRSctrl cells (shRNA control cells) to values comparable to those of IB3-1 or Caco-2/pRS26 cells (shRNA specific for CFTR). Treatments of IB3-1 or Caco-2/pRS26 cells with either IL-1β blocking antibody, IL-1 receptor antagonist, IKK inhibitor III (NF-KB pathway) or SB203580 (p38 MAPK pathway), restored the mCx-I activity. In addition, in IB3-1 or Caco-2/pRS26 cells, IL-1B blocking antibody, IKK inhibitor III or SB203580 reduced the mitochondrial ROS levels by ~50% and the cellular ROS levels near to basal values. The AP-1 inhibitors U0126 (MEK1/2) or SP600125 (JNK1/2/3 inhibitor) had no effects. The results suggest that in these cells IL-1B, through an autocrine effect, acts as a bridge connecting the CFTR with the mCx-I activity and the ROS levels.Fil: Clauzure, Mariangeles. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; ArgentinaFil: Valdivieso, Ángel Gabriel. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; ArgentinaFil: Massip Copiz, María Macarena. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; ArgentinaFil: Schulman, Gustavo. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; ArgentinaFil: Teiber, Maria Luz. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; ArgentinaFil: Tomás A. Santa-Coloma. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; Argentin

Response of mycorrhizal grapevine to Armillaria mellea inoculation: disease development and polyamines.

A study was conducted with the vine rootstock Richter 110 (Vitis berlandieri Planch. x Vitis rupestris L.) in order to assess whether the colonisation by the arbuscular mycorrhizal fungus (AMF) Glomus intraradices (BEG 72) can delay the disease development in plants inoculated with the root-rot fungus Armillaria mellea (Vahl:Fr) Kummer, and to elucidate if the levels of polyamines (PAs) are modified in response to G. intraradices, A. mellea or by the dual infection. Four treatments were considered: control and G. intraradices-inoculated plants infected or not with A. mellea. Plant growth, mycorrhizal colonisation and disease development were monitored throughout the experiment. High performance liquid chromatography (HPLC) in combination with fluorescence spectrophotometry was used to separate and quantify free root and leaf polyamines. The slower development of pathogenic symptoms and the higher plant biomass of mycorrhizal plants inoculated with A. mellea indicate an increase of tolerance due to the AMF inoculation. The variations in free PA levels detected at the beginning of the pathogenic infection suggest that PAs may have a potential role in the signalling mechanisms of the tolerance of mycorrhizal plants against A. mellea

The Role of Neurophysiology in Managing Patients with Chiari Malformations

Chiari type 1 malformation; Electromyography; SyringomyeliaMalformación de Chiari tipo 1; Electromiografía; SiringomieliaMalformació de Chiari tipus 1; Electromiografia; SiringomieliaChiari malformation type 1 (CM1) includes various congenital anomalies that share ectopia of the cerebellar tonsils lower than the foramen magnum, in some cases associated with syringomyelia or hydrocephalus. CM1 can cause dysfunction of the brainstem, spinal cord, and cranial nerves. This functional alteration of the nervous system can be detected by various modalities of neurophysiological tests, such as brainstem auditory evoked potentials, somatosensory evoked potentials, motor evoked potentials, electromyography and nerve conduction studies of the cranial nerves and spinal roots, as well as brainstem reflexes. The main goal of this study is to review the findings of multimodal neurophysiological examinations in published studies of patients with CM1 and their indication in the diagnosis, treatment, and follow-up of these patients, as well as their utility in intraoperative monitoring.This study was partially supported by grants FIS PI22/01082, which was co-financed by the European Regional Development Fund (ERDF), awarded to M.A. Poca and Agència de Gestió d’Ajuts Universitaris i de Recerca (AGAUR), Spain, grant 2021SGR/00810. The following nongovernmental associations have generously donated funding to support this research: (1) Asociación Nacional de Amigos de Arnold-Chiari (ANAC, http://www.arnoldchiari.es (accessed on 5 October 2023)), (2) Asociación Chiari y Siringomielia del Principado de Asturias (Ch.y.S.P.A., https://chyspa.org (accessed on 5 October 2023)), (3) Federación Española de Malformación de Chiari y Patologías Asociadas (FEMACPA), and (4) Mariana Dañobeitia https://vhir.vallhebron.com/es/sociedad/noticias/la-hija-de-una-paciente-con-malformacion-de-chiari-dona-21840-euros-para-impulsar-la-investigacion (accessed on 5 October 2023))

Nursing workload and staff allocation in an Italian hospital: a quality improvement initiative based on nursing care score

Aim: To develop, implement, and evaluate a Nursing Care Score (NCS) system, built into the electronic health record, to optimize nursing workload and staff allocation. Design: A quality improvement (QI) initiative with a pre- and post-implementation design was conducted by an interprofessional team in the 33-bed cardio-thoracic unit of a 72-bed hospital in Palermo, Italy. Methods: A seven-phase process was used to develop, implement, and evaluate the NCS, which lists 53 nursing work tasks, each assigned a score from 1.5 to 5.0. The nurse-to-patient ratio on all shifts was determined by the NCS. Nurse satisfaction with both the existing system and the NCS workload system was assessed. Descriptive statistics and McNemar's test were used to analyze the data. Results: At pre-implementation, 92.5% of nurses reported that the existing system was not effective, 87.5% reported it did not enable them to provide adequate nursing care, and 20.0% believed that workload was fairly distributed. At post-implementation, 75.0% of nurses reported that the NCS system was effective (p = 0.0348), 85.0% reported that the NCS system enabled them to provide adequate care, and 85.0% believed that workload was fairly distributed. An NCS score of 65 ± 5 was found to distribute workload most fairly. Conclusion: An automatic electronic operating system to generate a daily workload report based on the NCS was successfully implemented and evaluated. The NCS provided relevant information to guide nurse managers in defining nurse-to-patient ratio and determining staff allocation. Nurses were satisfied with the NCS system. The steps used to develop, implement, and evaluate the NCS system may be transferable to other units and other hospitals

El régimen natural de caudales: una diversidad imprescindible, una diversidad predecible

During last years the scientific community has realized flow regime importance as a connexion and explaining element of the fluvial ecosystem. Related to this paradigm, flow regime alteration analysis is necessary for the evaluation of environmental integrity. The objective is to show a new tool for characterizing stream flow regimes, quantifiying hydrologic impacts caused by dam construction. To reach this objective, this paper proposes: 1) to establish reference conditions and 2) evaluation of hydrological alteration: with 24 alteration indexes of alteration to measure the altered regime distortion against the natural. They have been applied to Vado reservoir situated in the upper reach of Jarama river which is integrated in Madrid water supply system.En los últimos años la comunidad científica ha puesto de manifiesto la importancia del régimen de caudales como elemento vertebrador del ecosistema fluvial, estableciendo el denominado «paradigma del régimen natural de caudales». A la luz de este paradigma, cualquier valoración de la integridad de un ecosistema fluvial debe incluir necesariamente un análisis objetivo de la alteración de su régimen de caudales. De hecho, la Directiva Marco del Agua establece el régimen hidrológico como uno de los elementos a considerar en la caracterización del estado ecológico de nuestros ríos. La metodología que se presenta en este trabajo permite: 1) definir el estatus hidrológico de referencia y 2) valorar la alteración del régimen de caudales, mediante 24 índices que evalúan la distorsión que en términos hidrológicos supone un determinado régimen frente al natural. Por último, se exponen los resultados de la aplicación de esta metodología al embalse de El Vado, ubicado en la cabecera del río Jarama e integrante del sistema de abastecimiento de agua a Madrid

Deep amplicon sequencing highlights low intra-host genetic variability of Echinococcus multilocularis and high prevalence of the European-type haplotypes in coyotes and red foxes in Alberta, Canada

Echinococcus multilocularis (Em) is a zoonotic parasite considered a global emergent pathogen. Recent findings indicate that the parasite is expanding its range in North America (NA) and that European-type (EU) haplotypes are circulating in western Canada. However, genetic analyses are usually conducted only on a few parasites out of thousands of individuals within each definitive host, likely underestimating the prevalence of less common haplotypes. Moreover, mixed infections with several mtDNA haplotypes in the same host have been reported, but their relative abundance within the host was never estimated. We aimed to 1) estimate the frequency of co-infections of different Em haplotypes in coyotes (Canis latrans) and red foxes (Vulpes vulpes) of western Canada and their relative abundance within the definitive hosts, 2) detect less prevalent haplotypes by sampling a larger proportion of the parasite subpopulation per host, and 3) investigate differences in the distribution of Em haplotypes in these main definitive hosts; foxes and coyotes. We extracted DNA from ~10% of the worm subpopulation per host (20 foxes and 47 coyotes) and used deep amplicon sequencing (NGS technology) on four loci, targeting the most polymorphic regions from the mitochondrial genes cox1 (814 bp), nad1 (344 bp), and cob (387 bp). We detected the presence of mixed infections with multiple Em haplotypes and with different Echinococcus species including E. granulosus s.l. genotypes G8/G10, low intraspecific diversity of Em, and a higher abundance of the EU-type haplotypes in both hosts. Our results suggest a population expansion of the European over the North American strain in Alberta and a limited distribution of some European-type haplotypes. Our findings indicate that deep amplicon sequencing represents a valuable tool to characterize Em in multiple hosts, to assess the current distribution and possible origins of the European strain in North America. The potential use of next-generation sequencing technologies is particularly important to understand the patterns of geographic expansion of this parasite

It’s a small world for parasites: evidence supporting the North American invasion of European Echinococcus multilocularis

Echinococcus multilocularis (Em), the causative agent of human alveolar echinococcosis (AE), is present in the Holarctic region as several genetic variants deemed to have differential infectivity and pathogenicity. An unprecedented outbreak of human AE cases in Western Canada infected with a European-like strain circulating in wild hosts warranted assessment of whether this strain was derived from a recent invasion or was endemic but undetected. Using nuclear and mitochondrial markers, we investigated the genetic diversity of Em in wild coyotes and red foxes from Western Canada, compared the genetic variants identified to global isolates, and assessed spatial distribution to infer possible invasion dynamics. Genetic variants from Western Canada were closely related to the original European clade, with lesser genetic diversity than that expected for a long-established strain and spatial genetic discontinuities within the study area, supporting the hypothesis of a relatively recent invasion with various founder events

Retromer deficiency in Tauopathy models enhances the truncation and toxicity of Tau.

Alteration of the levels, localization or post-translational processing of the microtubule associated protein Tau is associated with many neurodegenerative disorders. Here we develop adult-onset models for human Tau (hTau) toxicity in Drosophila that enable age-dependent quantitative measurement of central nervous system synapse loss and axonal degeneration, in addition to effects upon lifespan, to facilitate evaluation of factors that may contribute to Tau-dependent neurodegeneration. Using these models, we interrogate the interaction of hTau with the retromer complex, an evolutionarily conserved cargo-sorting protein assembly, whose reduced activity has been associated with both Parkinson’s and late onset Alzheimer’s disease. We reveal that reduction of retromer activity induces a potent enhancement of hTau toxicity upon synapse loss, axon retraction and lifespan through a specific increase in the production of a C-terminal truncated isoform of hTau. Our data establish a molecular and subcellular mechanism necessary and sufficient for the depletion of retromer activity to exacerbate Tau-dependent neurodegeneration.post-print2287 K

Linking Oxidative Events to Inflammatory and Adaptive Gene Expression Induced by Exposure to an Organic Particulate Matter Component

Background: Toxicological studies have correlated inflammatory effects of diesel exhaust particles (DEP) with its organic constituents, such as the organic electrophile 1,2-naphthoquinone (1,2-NQ)