The Expression of AQP1 IS Modified in Lung of Patients With Idiopathic Pulmonary Fibrosis: Addressing a Possible New Target

Activation of the epithelial-mesenchymal transition process (EMT) by which alveolar cells in human lung tissue undergo differentiation giving rise to a mesenchymal phenotype (fibroblast/miofibroblasts) has been well recognized as a key element in the origin of idiopathic pulmonary fibrosis (IPF). Here we analyzed expression of AQP1 in lung biopsies of patients diagnosed with IPF, and compared it to biopsies derived from patients with diverse lung pneumonies, such as hypersensitivity pneumonitis, sarcoidosis or normal lungs. Immunostaining for AQP1 showed a clear increment of AQP1 localized in the alveolar epithelium in biopsies from IPF patients alone. Moreover, to examine the possible participation of AQP1 in the pathophysiology of IPF, we evaluated its role in the pro-fibrotic transformation induced by transforming growth factor (TGF-β) in vitro. Human alveolar epithelial cells (A549), and fibroblasts derived from an IPF patient (LL29), or fibroblasts from healthy normal lung tissue (MRC-5), were treated with TGF-β, and levels of expression of AQP1, as well as those of E-cadherin, vimentin, α-SMA and collagen were analyzed by RT-qPCR, western blot and immunohistochemistry. An increase of AQP1 mRNA and protein after TGF-β treatment (4-72h) was observed either in A549 or IPF fibroblast-LL29 but not in MRC-5 fibroblasts. A gradual reduction of E-cadherin, and increased expression of vimentin, with no changes in α-SMA levels were observed in A549. Whereas in LL29 and MRC-5, TGF-β1 elicited a large production of collagen and α-SMA that was significantly greater in IPF fibroblast-LL29. Changes observed are consistent with activation of EMT by TGF-β, but whether modifications in AQP1 expression are responsible or independent events occurring at the same time is still unknown. Our results suggest that AQP1 plays a role in the pro-fibrotic TGF-β action and contributes to the etiology and pathophysiology of IPF. Understanding AQP1's role will help us comprehend the fate of this disease.Fondos FEDER (UE)Fundación NeumosurFundación SEPARISCIII Instituto de Salud Carlos II

Follow-up of health-related quality of life and pain in a cohort of patients with rheumatoid arthritis before and after COVID-19

The foot is one of the anatomical structures of the body most affected in rheumatoid arthritis (RA), associated with the disability of patients, even more during COVID-19. The aim of this study was to analyse whether the period of physical inactivity during COVID-19 is an influential factor on health-related quality of life and foot pain in patients with RA. Methods: 162 patients with foot pain and RA, recruited from the Hospital Virgen de las Nieves, Granada (Spain) were included. Data was collected during two different periods: January - December 2018 in person and June - September 2021 by phone. Patients were asked to complete the Spanish adapted version of the 12-Item Short Form Survey (SF-12) and the Visual Analogue Scale (VAS). Results: The results from the SF-12 questionnaires were divided between its two subscales (i.e., mental, and physical component). The physical component shows an improvement between 2018 and 2021, from 32.05in 2018–35.18 in 2021 (p < 0.05). The opposite happened with the mental component, showing a dete- rioration, from 39.69 in 2018–34.48 in 2021 (p < 0.05). Regarding pain, VAS shows higher levels of pain with statistically significant differences, both in general pain (from 6 in 2018–7 in 2012) and in foot pain (from 5 to 7), (p < 0.05). Conclusion: Mental quality of life and pain, both general and foot pain, are influenced by the period of physical inactivity during COVID-19.Funding for open access charge: Universidad de Málaga / CBU

Deciphering multiple sclerosis disability with deep learning attention maps on clinical MRI

Deep learning; Disability; Structural MRIAprendizaje profundo; Discapacidad; Resonancia magnética estructuralAprenentatge profund; Discapacitat; Ressonància magnètica estructuralThe application of convolutional neural networks (CNNs) to MRI data has emerged as a promising approach to achieving unprecedented levels of accuracy when predicting the course of neurological conditions, including multiple sclerosis, by means of extracting image features not detectable through conventional methods. Additionally, the study of CNN-derived attention maps, which indicate the most relevant anatomical features for CNN-based decisions, has the potential to uncover key disease mechanisms leading to disability accumulation. From a cohort of patients prospectively followed up after a first demyelinating attack, we selected those with T1-weighted and T2-FLAIR brain MRI sequences available for image analysis and a clinical assessment performed within the following six months (N = 319). Patients were divided into two groups according to expanded disability status scale (EDSS) score: ≥3.0 and < 3.0. A 3D-CNN model predicted the class using whole-brain MRI scans as input. A comparison with a logistic regression (LR) model using volumetric measurements as explanatory variables and a validation of the CNN model on an independent dataset with similar characteristics (N = 440) were also performed. The layer-wise relevance propagation method was used to obtain individual attention maps. The CNN model achieved a mean accuracy of 79% and proved to be superior to the equivalent LR-model (77%). Additionally, the model was successfully validated in the independent external cohort without any re-training (accuracy = 71%). Attention-map analyses revealed the predominant role of frontotemporal cortex and cerebellum for CNN decisions, suggesting that the mechanisms leading to disability accrual exceed the mere presence of brain lesions or atrophy and probably involve how damage is distributed in the central nervous system.MS PATHS is funded by Biogen. This study has been possible thanks to a Junior Leader La Caixa Fellowship awarded to C. Tur (fellowship code is LCF/BQ/PI20/11760008) by “la Caixa” Foundation (ID 100010434). The salaries of C. Tur and Ll. Coll are covered by this award

Early Pleistocene faunivorous hominins were not kleptoparasitic, and this impacted the evolution of human anatomy and socio-ecology

Humans are unique in their diet, physiology and socio-reproductive behavior compared to other primates. They are also unique in the ubiquitous adaptation to all biomes and habitats. From an evolutionary perspective, these trends seem to have started about two million years ago, coinciding with the emergence of encephalization, the reduction of the dental apparatus, the adoption of a fully terrestrial lifestyle, resulting in the emergence of the modern anatomical bauplan, the focalization of certain activities in the landscape, the use of stone tools, and the exit from Africa. It is in this period that clear taphonomic evidence of a switch in diet with respect to Pliocene hominins occurred, with the adoption of carnivory. Until now, the degree of carnivorism in early humans remained controversial. A persistent hypothesis is that hominins acquired meat irregularly (potentially as fallback food) and opportunistically through klepto-foraging. Here, we test this hypothesis and show, in contrast, that the butchery practices of early Pleistocene hominins (unveiled through systematic study of the patterning and intensity of cut marks on their prey) could not have resulted from having frequent secondary access to carcasses. We provide evidence of hominin primary access to animal resources and emphasize the role that meat played in their diets, their ecology and their anatomical evolution, ultimately resulting in the ecologically unrestricted terrestrial adaptation of our species. This has major implications to the evolution of human physiology and potentially for the evolution of the human brain

Role of aquaporins in cell proliferation: What else beyond water permeability?

In addition to the extensive data demonstrating the importance of mammalian AQPs for the movement of water and some small solutes across the cell membrane, there is now a growing body of evidence indicating the involvement of these proteins in numerous cellular processes seemingly unrelated, at least some of them in a direct way, to their canonical function of water permeation. Here, we have presented a broad range of evidence demonstrating that these proteins have a role in cell proliferation by various different mechanisms, namely, by allowing fast cell volume regulation during cell division; by affecting progression of cell cycle and helping maintain the balance between proliferation and apoptosis, and by crosstalk with other cell membrane proteins or transcription factors that, in turn, modulate progression of the cell cycle or regulate biosynthesis pathways of cell structural components. In the end, however, after discussing all these data that strongly support a role for AQPs in the cell proliferation process, it remains impossible to conclude that all these other functions attributed to AQPs occur completely independently of their water permeability, and there is a need for new experiments designed specifically to address this interesting issue.Carlos III Health Institute (Ref. PI12/01882)Regional Department of Health (PI 0018-2014

Cellular overexpression of Aquaporins slows down the natural HIF-2α degradation during prolonged hypoxia

Overexpression of cell membrane aquaporins (AQPs) has recently been associated with tumor formation, particularly with angiogenesis, cell migration and proliferation. Additionally, the hypoxia inducible factor (HIF) family has been extensively implicated in tumor growth and recent studies evidence interplay between AQP expression and HIF stability. Therefore, we decided to explore the effect that AQP overexpression has on the long-term stability of HIF-2α in PC12 cells exposed to chronic hypoxia, characteristic of a growing tumor. HIF-2α levels were analyzed in five PC12 clones with stable overexpression of different proteins (AQP1, AQP3, AQP5, G6PD, and GDNF), in PC12 transiently expressing G6PD or Kv4.2, and in wild-type PC12 cells. Overexpression of AQP1, 3 or 5 in PC12 cells (o-AQP-c) prevented the HIF-2α down-expression otherwise observed, after 16h at 1% O2, in wt-PC12 and in PC12 overexpressing non-AQP proteins. Longer HIF-2α stability was also observed in o-AQP-c exposed to cobalt chloride or dimethyloxallyl glycine. Normal proteasome activity was confirmed in all clones analyzed. Levels of HIF target genes (PHD2 and 3, VEGF, and PGK1) were 2-4 fold higher in hypoxic o-AQP-c than in wt-PC12 cells, and morphological changes in colony shape together with higher cell proliferation rates were observed in all o-AQP-c. Interestingly, analysis of PHD levels under normoxia revealed lower (50%) PHD3 expression in o-AQP-c than in controls. Our results indicate that AQP overexpression in PC12 cells prolongs HIF-2α stability during chronic hypoxia, leading to higher level of induction of its target genes and likely conferring to these cells a more tumor-like phenotype. © 2013 Elsevier B.V.This work was funded by grants from the “Instituto de Salud Carlos III” PS09/00605, and by grants from “La Junta de Andalucía”, Consejería de Salud (PI0298-2010) and Consejería de Innovación Ciencia y Empresa (P08-CTS-03574). AGC was supported by a predoctoral fellowship from “Junta de Andalucía, Consejería de Innovación Ciencia y Empresa”, Spain and IAR was supported by a predoctoral fellowship from Spanish Ministry of Science and Innovation.Peer Reviewe

Induction of cell apoptosis by nocodazole treatment.

<p><b>(A)</b> Flow cytometry dot plot analysis of annexin V/PI-stained cells to evaluate the induction of apoptosis by nocodazole (7.5μM, 24 h). (B) Summary of the percentage of cells found in each stage: annexin V^-/PI^- (alive), annexin V⁺/PI^- (apoptotic), annexin V⁺/PI⁺ (late apoptotic or dead) and annexin V^-/PI⁺ (necrotic). (C) Analysis by Western blot of levels of intact PARP protein (full) and cleaved PARP (upper panel) and summary of quantification analysis (bottom panel). Error bars are mean ± SEM from N experiments where N = 3 in (B) and N = 4 in (C). Significant differences are indicated as follows: *p ≤ 0.05, ** p ≤ 0.01 and *** p ≤ 0.001.</p

Analysis by flow cytometry of cell cycle profiles.

<p>(A). Cell cycle profiles showing the distribution of cells in the different phases of the cell cycle are presented. (B) Summary of the % of cells in each phase of the cycle. Significant differences (*** p ≤ 0.001) are indicated. (C) Western blot analysis of levels for cyclin E and D1 in each cell type (upper panel) and summary of the quantification analysis (bottom panel). The dash line at ratio of 1 indicates levels of cyclins in the wt cells. Bars are mean ± SEM from N experiments where N = 9 in (B) and N = 4 in (C).</p

Primer sequences used for the validation by qPCR amplification of different genes.

<p>Primer sequences used for the validation by qPCR amplification of different genes.</p

Cell cycle analysis after nocodazole treatment.

<p><b>(A)</b> The distribution of cells in the different phases of the cell cycle was analyzed in absence (control) or presence of nocodazole in both cell types, and a representative example of the cell cycle pattern after treatment with 7.5 μM of nocodazole is shown. (B) Summary of percentage of cells in each phase of the cell cycle with different concentrations (5, 7.5 and 10 μM, 24 h) of the drug. Significant differences are indicated as follows: * p ≤ 0.05, ** p ≤ 0.01 and *** p ≤ 0.001. Bars are mean ± SEM from 4–6 experiments.</p