Investigation on lysosomal accumulation by a quantitative analysis of 2D phase-maps in digital holography microscopy.

Lysosomes are the terminal end of catabolic pathways in the cell, as well as signaling centers performing important functions such as the recycling of macromolecules, organelles, and nutrient adaptation. The importance of lysosomes in human health is supported by the fact that the deficiency of most lysosomal genes causes monogenic diseases called as a group Lysosomal Storage Diseases (LSDs). A common phenotypic hallmark of LSDs is the expansion of the lysosomal compartment that can be detected by using conventional imaging methods based on immunofluorescence protocols or overexpression of tagged lysosomal proteins. These methods require the alteration of the cellular architecture (i.e., due to fixation methods), can alter the behavior of cells (i.e., by the overexpression of proteins), and require sample preparation and the accurate selection of compatible fluorescent markers in relation to the type of analysis, therefore limiting the possibility of characterizing cellular status with simplicity. Therefore, a quantitative and label-free methodology, such as Quantitative Phase Imaging through Digital Holographic (QPI-DH), for the microscopic imaging of lysosomes in health and disease conditions may represent an important advance to study and effectively diagnose the presence of lysosomal storage in human disease. Here we proof the effectiveness of the QPI-DH method in accomplishing the detection of the lysosomal compartment using mouse embryonic fibroblasts (MEFs) derived from a Mucopolysaccharidosis type III-A (MSP-IIIA) mouse model, and comparing them with wild-type (WT) MEFs. We found that it is possible to identify label-free biomarkers able to supply a first pre-screening of the two populations, thus showing that QPI-DH can be a suitable candidate to surpass fluorescent drawbacks in the detection of lysosomes dysfunction. An appropriate numerical procedure was developed for detecting and evaluate such cellular substructures from in vitro cells cultures. Results reported in this study are encouraging about the further development of the proposed QPI-DH approach for such type of investigations about LSDs.

Activation of Ca2+ phosphatase Calcineurin regulates Parkin translocation to mitochondria and mitophagy in flies.

Selective removal of dysfunctional mitochondria via autophagy is crucial for the maintenance of cellular homeostasis. This event is initiated by the translocation of the E3 ubiquitin ligase Parkin to damaged mitochondria, and it requires the Serine/Threonine-protein kinase PINK1. In a coordinated set of events, PINK1 operates upstream of Parkin in a linear pathway that leads to the phosphorylation of Parkin, Ubiquitin, and Parkin mitochondrial substrates, to promote ubiquitination of outer mitochondrial membrane proteins. Ubiquitin-decorated mitochondria are selectively recruiting autophagy receptors, which are required to terminate the organelle via autophagy. In this work, we show a previously uncharacterized molecular pathway that correlates the activation of the Ca2+-dependent phosphatase Calcineurin to Parkin translocation and Parkin-dependent mitophagy. Calcineurin downregulation or genetic inhibition prevents Parkin translocation to CCCP-treated mitochondria and impairs stress-induced mitophagy, whereas Calcineurin activation promotes Parkin mitochondrial recruitment and basal mitophagy. Calcineurin interacts with Parkin, and promotes Parkin translocation in the absence of PINK1, but requires PINK1 expression to execute mitophagy in MEF cells. Genetic activation of Calcineurin in vivo boosts basal mitophagy in neurons and corrects locomotor dysfunction and mitochondrial respiratory defects of a Drosophila model of impaired mitochondrial functions. Our study identifies Calcineurin as a novel key player in the regulation of Parkin translocation and mitophagy.

Enfermedades pulmonares intersticiales difusas asociadas al tabaquismo / Diffuse interstitial lung diseases associated with smoking

El interrogatorio sobre hábito tabáquico no debe obviarse jamás, pues el abanico de posibilidades diagnósticas inherentes es amplio. Es un gesto clínico fundamental frente a individuos que consultan por síntomas y/o hallazgos radiológicos casuales. Presentamos tres casos clínicos: síndrome de combinación de fibrosis pulmonar y enfisema, neumonitis intersticial descamativa e histiocitosis de células de Langerhans, como parte del abanico de las enfermedades pulmonares intersticiales difusas asociadas a tabaco (EPIDAT), donde la tomografía de alta resolución de tórax tiene un rol destacado. Palabras clave: tabaquismo; enfisema; enfermedades pulmonares intersticiales; histiocitosis de células de Langerhans

Questioning about smoking habits should never be ignored, since the range of inherent diagnostic possibilities is wide. It is a fundamental clinical step facing individuals who consult for symptoms and/or casual radiological findings. We present three clinical cases: the combined syndrome of pulmonary fibrosis and emphysema, desquamative interstitial pneumonitis, and Langerhans cell histiocytosis, as part of the range of tobacco-associated diffuse interstitial lung diseases (EPIDAT), where high-resolution chest tomography has a prominent role. Key words: smoking; emphysema; interstitial lung diseases; Langerhans cell histiocytosis

Eutrombicula cochinocaensis infestation in llamas (Lama glama) in the Puna region of Argentina.

This paper aims to describe a case of trombiculosis in llamas from the Puna region of Argentina caused by Eutrombicula cochinocaensis. Trombiculosis is a parasitic infestation caused by the larval form of trombiculid mites, commonly known as chigger mites. Six adult llamas out of eighteen (33%) were infested and showed dermatitis, hyperemia, and crusts together with an orange-dotted pattern. The chigger mites were found mainly in the hind limbs and the abdomen. Microscopic identification of E. cochinocaensis was made by phase-contrast microscopy. Then, a histopathological study of the affected animals' skin was performed. Histological findings included dermatitis withmainly lymphocytic infiltrate, pustules, crusts, hyperkeratosis, and the mites' stylostomes in the dermis and epidermis. This is the first report of trombiculosis caused by E. cochinocaensis in llamas from Argentina. Mites of the family Trombiculidae are vectors of certain diseases to humans and animals and are therefore of sanitary and productive importance.

miR-218 Promotes Dopaminergic Differentiation and Controls Neuron Excitability and Neurotransmitter Release through the Regulation of a Synaptic-Related Genes Network.

In the brain, microRNAs (miRNAs) are believed to play a role in orchestrating synaptic plasticity at a higher level by acting as an additional mechanism of translational regulation, alongside the mRNA/polysome system. Despite extensive research, our understanding of the specific contribution of individual miRNA to the function of dopaminergic neurons (DAn) remains limited. By performing a dopaminergic-specific miRNA screening, we have identified miR-218 as a critical regulator of DAn activity in male and female mice. We have found that miR-218 is specifically expressed in mesencephalic DAn and is able to promote dopaminergic differentiation of embryonic stem cells and functional maturation of transdifferentiated induced DA neurons. Midbrain-specific deletion of both genes encoding for miR-218 (referred to as miR-218-1 and mir218-2) affects the expression of a cluster of synaptic-related mRNAs and alters the intrinsic excitability of DAn, as it increases instantaneous frequencies of evoked action potentials, reduces rheobase current, affects the ionic current underlying the action potential after hyperpolarization phase, and reduces dopamine efflux in response to a single electrical stimulus. Our findings provide a comprehensive understanding of the involvement of miR-218 in the dopaminergic system and highlight its role as a modulator of dopaminergic transmission.SIGNIFICANCE STATEMENT In the past decade, several miRNAs have emerged as potential regulators of synapse activity through the modulation of specific gene expression. Among these, we have identified a dopaminergic-specific miRNA, miR-218, which is able to promote dopaminergic differentiation and regulates the translation of an entire cluster of synapse related mRNAs. Deletion of miR-218 has notable effects on dopamine release and alters the intrinsic excitability of dopaminergic neurons, indicating a direct control of dopaminergic activity by miR-218.

[Post-colonoscopy colorectal cancer: Evaluation of a cohort in its clinical and colonoscopic characteristics, survival and its causes according to the World Endoscopy Organization]. / Cáncer colorrectal post-colonoscopia: Evaluación de una cohorte en sus características clínicas, colonoscópicas, sobrevida y sus causas según la Organización Mundial de Endoscopia.

Post-colonoscopy colorectal cancer (PCCRC) is a tumor that appears after a normal colonoscopy before the established time for the endoscopic follow up. Its origin reflects the quality of the colonoscopy and the different tumoral biologics between the CRC and the CRCPC. Our aim is to describe the characteristics of the PCCRC in our region, to identify risk factors, to discriminate the potential causes according to the World Endoscopý Organization (WEO) and to determine its impact in the patient's survival. We studied patients with colorectal cancer (CRC) attended at the gastro-oncology clinic of two institutions of Medellin-Colombia, between January 2012 and December 2021 that had been submitted to a colonoscopy between 6-36 months before the colonoscopy in which the CRC was diagnosed. 919 patients during 10 years for CRC, 68 cases of PCCRC (6.9%); It was more frequent in older patients (74 vs. 66 years; p=0.03), with background of adenomatous polyps (36.8% vs. 20.1%; p=0.01) and in right colon (57.4% vs. 40.6%; p=0.006), with a tendency in patients with diverticulosis (41.2% vs. 31.3%; p=0.05) and diabetes (25% vs. 14%; p=0.06); less survival at 5 and 10 years (58% and 55.2% vs. 67% and 63%; p < 0.001). According to the WEO, the PCCRC presents in 61.3% because of abnormal findings omitted in inadequate colonoscopies, 29% in a suitable colonoscopy and 9.7% incomplete resections of adenomas. In conclusion, the rate of PCCRC was 6.9% with more propension in older patients, a background of polyp resection, and proximal colon. According to the WEO, the abnormal findings omitted more frequently were related with inadequate colonoscopies. The patients with PCCRC had less survival.

Loss of the batten disease protein CLN3 leads to mis-trafficking of M6PR and defective autophagic-lysosomal reformation.

Batten disease, one of the most devastating types of neurodegenerative lysosomal storage disorders, is caused by mutations in CLN3. Here, we show that CLN3 is a vesicular trafficking hub connecting the Golgi and lysosome compartments. Proteomic analysis reveals that CLN3 interacts with several endo-lysosomal trafficking proteins, including the cation-independent mannose 6 phosphate receptor (CI-M6PR), which coordinates the targeting of lysosomal enzymes to lysosomes. CLN3 depletion results in mis-trafficking of CI-M6PR, mis-sorting of lysosomal enzymes, and defective autophagic lysosomal reformation. Conversely, CLN3 overexpression promotes the formation of multiple lysosomal tubules, which are autophagy and CI-M6PR-dependent, generating newly formed proto-lysosomes. Together, our findings reveal that CLN3 functions as a link between the M6P-dependent trafficking of lysosomal enzymes and lysosomal reformation pathway, explaining the global impairment of lysosomal function in Batten disease.

Cáncer colorrectal post-colonoscopia: Evaluación de una cohorte en sus características clínicas, colonoscópicas, sobrevida y sus causas según la Organización Mundial de Endoscopia

Cáncer colorrectal post-colonoscopia (CCRP) es el tumor que aparece posterior a una colonoscopia normal antes de cumplirse el tiempo establecido para seguimiento endoscópico. Origen multifactorial, refleja la calidad de la colonoscopia y las diferentes biologías tumorales entre los cánceres colorrectales detectados (CCRD) y el CCRP. Nuestro objetivo es describir las características del CCRP en nuestro medio, identificar factores de riesgo, discriminar sus causas según la Organización Mundial de Endoscopia (OME) y determinar el efecto en la sobrevida del paciente. El estudio se realizó en pacientes con cáncer-colorrectal (CCR) atendidos en consulta de gastro-oncología de dos instituciones en Medellín-Colombia, entre enero de 2012 y diciembre de 2021 que se habían sometido a una colonoscopia en los 6 a 36 meses anteriores a la colonoscopia en la que se diagnosticó el CCR. 919 pacientes durante 10 años por CCR, 68 casos de CCRP (6,9%), se encontró que se presenta con más frecuencia en pacientes mayores (74 vs. 66 años; p=0,03), con antecedentes de pólipos adenomatosos (36,8% vs. 20,1%; p=0,01) y en colon derecho (57,4% vs. 40,6%; p=0,006), con una tendencia en pacientes con diverticulosis (41,2% vs. 31,3%; p=0,05) y diabetes (25% vs. 14%; p=0,06); menor sobrevida a 5 y 10 años (58%-55,2% vs. 67%-63%; p<0,001). Según la OME, los CCRP se presentaron en 61,3% por lesiones omitidas en colonoscopias inadecuadas, 29% colonoscopias adecuadas y 9,7% resecciones incompletas de adenomas. En conclusión, la tasa de CCRP fue de 6,9%, con mayor propensión en pacientes de mayores, antecedente de resección de pólipos, y en colon derecho. Acorde a la OME, las lesiones omitidas más frecuentemente se relacionaron con colonoscopias inadecuadas. Los pacientes con CCRP tienen menor sobrevida.

Post-colonoscopy colorectal cancer (PCCRC) is a tumor that appears after a normal colonoscopy before the established time for the endoscopic follow up. Its origin reflects the quality of the colonoscopy and the different tumoral biologics between the CRC and the CRCPC. Our aim is to describe the characteristics of the PCCRC in our region, to identify risk factors, to discriminate the potential causes according to the World Endoscopý Organization (WEO) and to determine its impact in the patient's survival. We studied patients with colorectal cancer (CRC) attended at the gastro-oncology clinic of two institutions of Medellin-Colombia, between January 2012 and December 2021 that had been submitted to a colonoscopy between 6-36 months before the colonoscopy in which the CRC was diagnosed. 919 patients during 10 years for CRC, 68 cases of PCCRC (6.9%); It was more frequent in older patients (74 vs. 66 years; p=0.03), with background of adenomatous polyps (36.8% vs. 20.1%; p=0.01) and in right colon (57.4% vs. 40.6%; p=0.006), with a tendency in patients with diverticulosis (41.2% vs. 31.3%; p=0.05) and diabetes (25% vs. 14%; p=0.06); less survival at 5 and 10 years (58% and 55.2% vs. 67% and 63%; p<0.001). According to the WEO, the PCCRC presents in 61.3% because of abnormal findings omitted in inadequate colonoscopies, 29% in a suitable colonoscopy and 9.7% incomplete resections of adenomas. In conclusion, the rate of PCCRC was 6.9% with more propension in older patients, a background of polyp resection, and proximal colon. According to the WEO, the abnormal findings omitted more frequently were related with inadequate colonoscopies. The patients with PCCRC had less survival.

EGR1 drives cell proliferation by directly stimulating TFEB transcription in response to starvation.

The stress-responsive transcription factor EB (TFEB) is a master controller of lysosomal biogenesis and autophagy and plays a major role in several cancer-associated diseases. TFEB is regulated at the posttranslational level by the nutrient-sensitive kinase complex mTORC1. However, little is known about the regulation of TFEB transcription. Here, through integrative genomic approaches, we identify the immediate-early gene EGR1 as a positive transcriptional regulator of TFEB expression in human cells and demonstrate that, in the absence of EGR1, TFEB-mediated transcriptional response to starvation is impaired. Remarkably, both genetic and pharmacological inhibition of EGR1, using the MEK1/2 inhibitor Trametinib, significantly reduced the proliferation of 2D and 3D cultures of cells displaying constitutive activation of TFEB, including those from a patient with Birt-Hogg-Dubé (BHD) syndrome, a TFEB-driven inherited cancer condition. Overall, we uncover an additional layer of TFEB regulation consisting in modulating its transcription via EGR1 and propose that interfering with the EGR1-TFEB axis may represent a therapeutic strategy to counteract constitutive TFEB activation in cancer-associated conditions.

Preventable risk factors for type 2 diabetes can be detected using noninvasive spontaneous electroretinogram signals.

Given the ever-increasing prevalence of type 2 diabetes and obesity, the pressure on global healthcare is expected to be colossal, especially in terms of blindness. Electroretinogram (ERG) has long been perceived as a first-use technique for diagnosing eye diseases, and some studies suggested its use for preventable risk factors of type 2 diabetes and thereby diabetic retinopathy (DR). Here, we show that in a non-evoked mode, ERG signals contain spontaneous oscillations that predict disease cases in rodent models of obesity and in people with overweight, obesity, and metabolic syndrome but not yet diabetes, using one single random forest-based model. Classification performance was both internally and externally validated, and correlation analysis showed that the spontaneous oscillations of the non-evoked ERG are altered before oscillatory potentials, which are the current gold-standard for early DR. Principal component and discriminant analysis suggested that the slow frequency (0.4-0.7 Hz) components are the main discriminators for our predictive model. In addition, we established that the optimal conditions to record these informative signals, are 5-minute duration recordings under daylight conditions, using any ERG sensors, including ones working with portative, non-mydriatic devices. Our study provides an early warning system with promising applications for prevention, monitoring and even the development of new therapies against type 2 diabetes.

Antagonistic effect of cyclin-dependent kinases and a calcium-dependent phosphatase on polyglutamine-expanded androgen receptor toxic gain of function.

Spinal and bulbar muscular atrophy is caused by polyglutamine (polyQ) expansions in androgen receptor (AR), generating gain-of-function toxicity that may involve phosphorylation. Using cellular and animal models, we investigated what kinases and phosphatases target polyQ-expanded AR, whether polyQ expansions modify AR phosphorylation, and how this contributes to neurodegeneration. Mass spectrometry showed that polyQ expansions preserve native phosphorylation and increase phosphorylation at conserved sites controlling AR stability and transactivation. In small-molecule screening, we identified that CDC25/CDK2 signaling could enhance AR phosphorylation, and the calcium-sensitive phosphatase calcineurin had opposite effects. Pharmacologic and genetic manipulation of these kinases and phosphatases modified polyQ-expanded AR function and toxicity in cells, flies, and mice. Ablation of CDK2 reduced AR phosphorylation in the brainstem and restored expression of Myc and other genes involved in DNA damage, senescence, and apoptosis, indicating that the cell cycle-regulated kinase plays more than a bystander role in SBMA-vulnerable postmitotic cells.

Soberanía alimentaria y salud: perspectivas de tres pueblos indígenas de Colombia.

RESUMEN: La soberanía alimentaria es el derecho colectivo a decidir sobre la producción, distribución y consumo de alimentos y promueve la generación de suministros alimenticios para el consumo local de tal forma que los consumidores queden resguardados de la volatilidad de los precios de mercados internacionales. En los pueblos indígenas esta tendencia resulta trascendental para hacer frente a condiciones de inequidad histórica que han impactado negativamente su salud pública. No obstante, es ignorada en algunos casos y en otros reemplazada por nociones centradas en la seguridad alimentaria, término alineado con políticas transnacionales derivadas del modelo económico dominante. OBJETIVO: analizar con tres comunidades indígenas de Colombia las perspectivas de soberanía alimentaria y su influencia en la salud. MÉTODO: investigación participativa basada en la comunidad, para recolectar datos mediante grupos de discusión, entrevistas y observación. La población de estudio fueron tres comunidades indígenas del sur de Colombia. La selección de participantes se realizó según su trayectoria en la comunidad. RESULTADOS: las comunidades entienden la soberanía alimentaria como la conservación de semillas nativas y alimentos propios, vista como oportunidad para el cuidado de la salud. Su debilitamiento se relaciona con el desarrollo de enfermedades de las personas y de la madre tierra. Para su fortalecimiento identificaron el tul, el yatul y la chagra (huerta) que reafirman la unión familiar, contribuyen a la recuperación de modos de producción desde la sabiduría ancestral y se posicionan como alternativa para la sostenibilidad económica. CONCLUSIÓN: la soberanía alimentaria conserva los saberes y prácticas tradicionales para una alimentación propia, debilitada por los sistemas agroindustriales. Es asumida como iniciativa local suscrita en un proyecto global de resistencia política y económica para la salud colectiva de los pueblos.

TRPML1 and TFEB, an Intimate Affair.

Ca2+ is a universal second messenger that plays a wide variety of fundamental roles in cellular physiology. Thus, to warrant selective responses and to allow rapid mobilization upon specific stimuli, Ca2+ is accumulated in organelles to keep it at very low levels in the cytoplasm during resting conditions. Major Ca2+ storage organelles include the endoplasmic reticulum (ER), the mitochondria, and as recently demonstrated, the lysosome (Xu and Ren, Annu Rev Physiol 77:57-80, 2015). The importance of Ca2+ signaling deregulation in human physiology is underscored by its involvement in several human diseases, including lysosomal storage disorders, neurodegenerative disease and cancer (Shen et al., Nat Commun 3:731, 2012; Bae et al., J Neurosci 34:11485-11503, 2014). Recent evidence strongly suggests that lysosomal Ca2+ plays a major role in the regulation of lysosomal adaptation to nutrient availability through a lysosomal signaling pathway involving the lysosomal Ca2+ channel TRPML1 and the transcription factor TFEB, a master regulator for lysosomal function and autophagy (Sardiello et al., Science 325:473-477, 2009; Settembre et al., Science 332:1429-1433, 2011; Medina et al., Nat Cell Biol 17:288-299, 2015; Di Paola et al., Cell Calcium 69:112-121, 2018). Due to the tight relationship of this lysosomal Ca2+ channel and TFEB, in this chapter, we will focus on the role of the TRPML1/TFEB pathway in the regulation of lysosomal function and autophagy.

COVID-19: una oportunidad para revitalizar la autonomía alimentaria en comunidades indígenas / COVID-19: an opportunity to revitalize food autonomy in indigenous communities / Covid-19: uma oportunidade para revitalizar a autonomia alimentar nas comunidades indígenas

Resumen La autonomía alimentaria permite a las comunidades determinar sus prácticas alimentarias, comenzando con la producción agrícola en sus territorios garantizando la economía propia y la armonía con la madre tierra, en este contexto, la pandemia por COVID-19 generó cambios en las practicas sociales en las comunidades indígenas, incluidas sus prácticas alimentarias. Objetivo: Describir prácticas de autonomía alimentaria revitalizadas a partir del confinamiento por COVID-19 en una comunidad indígena Nasa del sur de Colombia. Método: Estudio cualitativo orientado en investigación participativa basada en comunidad (CBPR) realizado en una comunidad indígena Nasa del sur de Colombia. La recolección e interpretación de información se efectuó con técnicas de la investigación cualitativa: codificación abierta y axial y hasta llegar a la descripción de categorías emergentes. Resultados: Se fortalecieron prácticas ancestrales del sistema alimentario para mejorar el acceso a los alimentos sanos producidos en sus territorios, dentro de las que se destacan: tul (huerta familiar), trueque, mano-cambio, mercado Nasa y recetas tradicionales. Conclusiones: El confinamiento representó una oportunidad para retomar y revitalizar las prácticas alimentarias ancestrales de la comunidad que respondieron a necesidades concretas de salud y de alimentación, fortaleciendo el tejido social y la identidad indígena, acciones que pueden trascender a políticas públicas, planes de vida y aspiraciones de buen vivir.

Abstract Food autonomy allows communities to determine their food practices, starting with agricultural production in their territories, guaranteeing their own economy and harmony with Mother Earth. In this context, the COVID-19 pandemic generated changes in social practices in the communities. indigenous communities, including their food practices. Objective: Describe food autonomy practices revitalized following the COVID-19 confinement in a Nasa indigenous community in southern Colombia. Method: Qualitative study oriented on community-based participatory research (CBPR) carried out in a Nasa indigenous community in southern Colombia. The collection and interpretation of information was carried out with qualitative research techniques: open and axial coding and until reaching the description of emerging categories. Results: Ancestral practices of the food system were strengthened to improve access to healthy foods produced in their territories, among which the following stand out: tul (family garden), barter, hand-exchange, Nasa market and traditional recipes. Conclusions: Confinement represented an opportunity to resume and revitalize the community's ancestral food practices that responded to specific health and food needs, strengthening the social fabric and indigenous identity, actions that can transcend public policies, life plans and aspirations for a good life.

Abstract A autonomia alimentar permite que as comunidades determinem suas práticas alimentares, começando pela produção agrícola em seus territórios, garantindo sua própria economia e harmonia com a Mãe Terra. Nesse contexto, a pandemia da Covid-19 gerou mudanças nas práticas sociais nas comunidades indígenas, incluindo suas práticas alimentares. Objetivo: Descrever as práticas de autonomia alimentar revitalizadas após o confinamento da COVID-19 em uma comunidade indígena Nasa no sul da Colômbia. Método: Estudo qualitativo orientado à pesquisa participativa de base comunitária (CBPR) realizada em uma comunidade indígena Nasa no sul da Colômbia. A coleta e interpretação das informações foram realizadas com técnicas de pesquisa qualitativa: codificação aberta e axial e até chegar à descrição das categorias emergentes. Resultados: As práticas ancestrais do sistema alimentar foram fortalecidas para melhorar o acesso aos alimentos saudáveis produzidos em seus territórios, entre os quais se destacam: tul (horta familiar), escambo, troca de mãos, mercado Nasa e receitas tradicionais. Conclusões: O confinamento representou uma oportunidade para retomar e revitalizar as práticas alimentares ancestrais da comunidade que respondiam às necessidades específicas de saúde e alimentação, fortalecendo o tecido social e a identidade indígena, ações que podem transcender as políticas públicas, os planos de vida e as aspirações por uma vida boa.

Lysosomal fusion and SNARE function are impaired by cholesterol accumulation in lysosomal storage disorders.

Partial Retraction of: The EMBO Journal (2010) 29: 3607-3620. DOI: 10.1038/emboj.2010.237 | Published online 24 September 2010 Journal statement The journal contacted the authors in February 2022 about potential image insertions and duplications in Fig 4A and 4E. In the absence of source data, the authors are retracting Fig 4A, the lower panel of Fig 4E (LAMP1 immunoblot), and the following statements in the text that rely on these data: "Quantitative analysis showed that the percentage of Flotillin-1 associated with DRMs was increased in LSD endolysosomal membranes (Figure 4A), indicating an increased amount of cholesterol-enriched regions in these membrane samples." "LAMP1 also displayed a similar distribution profile in WT and LSD cells (Figure 4E)". Author statement The authors could not verify the aberrations in panel A of Fig 4 and the lower immunoblot (LAMP1) of 4E because the original source data are no longer available (12 years after publication, which is beyond the institute's 10-year data retention policy). The authors wish to clarify that the main conclusions of the paper are not affected by the retraction of Figure panels 4A and 4E for the following reasons: Figure panel 4A supports the observation that there are increased cholesterol-enhanced regions in LSD samples. This finding is also supported by data provided in figs 4B, 4C and 4D. Figure panel 4E: The LAMP1 blot in Fig 4E shows that the distribution of protein normally excluded from DRMs is not altered between Wt and LSD samples. This result is also supported by the upper blot in this panel (Transferrin receptor). The authors apologize for these errors and agree with this corrigendum; no response could be obtained from AL.

Recomendaciones de la Sociedad Española de Cirugía Torácica para el manejo del derrame pleural maligno / Recommendations of the Spanish Society of Thoracic Surgery for the management of malignant pleural effusion

Este artículo resume la guía clínica de diagnóstico y tratamiento del derrame pleural maligno (DPM) auspiciada por la Sociedad Española de Cirugía Torácica (SECT). Se elaboraron 10 controversias clínicas bajo la metodología de preguntas PICO (Patient, Intervention, Comparison, Outcome) y la calidad de la evidencia y graduación de la fuerza de las recomendaciones se basó en el sistema Grading of Recommendations, Assessment, Development and Evaluations (GRADE). El análisis inmunocitoquímico y molecular del líquido pleural puede evitar procedimientos invasivos ulteriores con finalidad diagnóstica. Actualmente, el control definitivo del DPM se puede realizar indistintamente a través de una pleurodesis (talco poudrage o slurry) o de la inserción de un catéter pleural tunelizado (CPT). Es probable que la combinación de ambas técnicas (p.ej. toracoscopia con talco poudrage e inserción de un CPT, o instilación de talco slurry a través de un CPT) ocupe un lugar predominante en el manejo terapéutico futuro. (AU)

This article summarizes the clinical guidelines for the diagnosis and treatment of malignant pleural effusion (MPE) sponsored by the Spanish Society of Thoracic Surgery (SECT). Ten clinical controversies were elaborated under the methodology of PICO (Patient, Intervention, Comparison, Outcome) questions and the quality of the evidence and grading of the strength of the recommendations was based on the GRADE system. Immunocytochemical and molecular analyses of pleural fluid may avoid further invasive diagnostic procedures. Currently, the definitive control of MPE can be achieved either by pleurodesis (talc poudrage or slurry) or the insertion of a indwelling pleural catheter (IPC). It is likely that the combination of both techniques (i.e., thoracoscopy with talc poudrage and insertion of a IPC, or instillation of talc slurry through a IPC) will have a predominant role in the future therapeutic management. (AU)

TPC2 rescues lysosomal storage in mucolipidosis type IV, Niemann-Pick type C1, and Batten disease.

Lysosomes are cell organelles that degrade macromolecules to recycle their components. If lysosomal degradative function is impaired, e.g., due to mutations in lysosomal enzymes or membrane proteins, lysosomal storage diseases (LSDs) can develop. LSDs manifest often with neurodegenerative symptoms, typically starting in early childhood, and going along with a strongly reduced life expectancy and quality of life. We show here that small molecule activation of the Ca2+ -permeable endolysosomal two-pore channel 2 (TPC2) results in an amelioration of cellular phenotypes associated with LSDs such as cholesterol or lipofuscin accumulation, or the formation of abnormal vacuoles seen by electron microscopy. Rescue effects by TPC2 activation, which promotes lysosomal exocytosis and autophagy, were assessed in mucolipidosis type IV (MLIV), Niemann-Pick type C1, and Batten disease patient fibroblasts, and in neurons derived from newly generated isogenic human iPSC models for MLIV and Batten disease. For in vivo proof of concept, we tested TPC2 activation in the MLIV mouse model. In sum, our data suggest that TPC2 is a promising target for the treatment of different types of LSDs, both in vitro and in-vivo.

The Regulation of MiTF/TFE Transcription Factors Across Model Organisms: from Brain Physiology to Implication for Neurodegeneration.

The microphthalmia/transcription factor E (MiTF/TFE) transcription factors are responsible for the regulation of various key processes for the maintenance of brain function, including autophagy-lysosomal pathway, lipid catabolism, and mitochondrial homeostasis. Among them, autophagy is one of the most relevant pathways in this frame; it is evolutionary conserved and crucial for cellular homeostasis. The dysregulation of MiTF/TFE proteins was shown to be involved in the development and progression of neurodegenerative diseases. Thus, the characterization of their function is key in the understanding of the etiology of these diseases, with the potential to develop novel therapeutics targeted to MiTF/TFE proteins and to the autophagic process. The fact that these proteins are evolutionary conserved suggests that their function and dysfunction can be investigated in model organisms with a simpler nervous system than the mammalian one. Building not only on studies in mammalian models but also in complementary model organisms, in this review we discuss (1) the mechanistic regulation of MiTF/TFE transcription factors; (2) their roles in different regions of the central nervous system, in different cell types, and their involvement in the development of neurodegenerative diseases, including lysosomal storage disorders; (3) the overlap and the compensation that occur among the different members of the family; (4) the importance of the evolutionary conservation of these protein and the process they regulate, which allows their study in different model organisms; and (5) their possible role as therapeutic targets in neurodegeneration.

Risk factors and incidence of cytomegalovirus viremia and disease in pediatric patients with allogeneic hematopoietic stem cell transplantation: An 8-year single-center experience in Latin America.

BACKGROUND: Cytomegalovirus infection represents a significant cause of morbidity and mortality after hematopoietic stem cell transplantation. This study aimed to evaluate the incidence of viremia and disease due to cytomegalovirus and the risk factors in pediatric patients with hematopoietic stem cell transplantation in our institution. METHODS: This was a retrospective cohort of patients under 19 years of age who underwent allogeneic hematopoietic stem cell transplantation due to any indication between 2012 and 2019. The analysis included the diagnosis of cytomegalovirus viremia or disease during post-transplant follow-up, evaluation of risk factors, and outcomes. The statistical analysis included univariate and multivariate analyses, and the cumulative incidence of cytomegalovirus viremia was determined by the Kaplan-Meier method using STATA 14 statistical software. RESULTS: A total of 182 transplants were included. At 100 days, the cumulative incidence of cytomegalovirus viremia was 70.5%, and that of cytomegalovirus disease was 4.7%. Overall survival at 2 years was 74%, and event-free survival was 64%. The remaining demographic characteristics were not predictors of infection. There was no association between viremia and relapse or survival of the patients. Higher mortality was noted in cytomegalovirus disease. CONCLUSIONS: During the study period, the incidence of cytomegalovirus disease was similar to that of other pediatric reports, but the incidence of viremia was higher. Pre-emptive therapy has diminished disease rates and death due to infection. Viral load cutoff points should be standardized to guide treatment and avoid myelotoxicity.