In vivo Polycystin-1 interactome using a novel Pkd1 knock-in mouse model.

PKD1 is the most commonly mutated gene causing autosomal dominant polycystic kidney disease (ADPKD). It encodes Polycystin-1 (PC1), a putative membrane protein that undergoes a set of incompletely characterized post-transcriptional cleavage steps and has been reported to localize in multiple subcellular locations, including the primary cilium and mitochondria. However, direct visualization of PC1 and detailed characterization of its binding partners remain challenging. We now report a new mouse model with HA epitopes and eGFP knocked-in frame into the endogenous mouse Pkd1 gene by CRISPR/Cas9. Using this model, we sought to visualize endogenous PC1-eGFP and performed affinity-purification mass spectrometry (AP-MS) and network analyses. We show that the modified Pkd1 allele is fully functional but the eGFP-tagged protein cannot be detected without signal amplification by secondary antibodies. Using nanobody-coupled beads and large quantities of tissue, AP-MS identified an in vivo PC1 interactome, which is enriched for mitochondrial proteins and components of metabolic pathways. These studies suggest this mouse model and interactome data will be useful to understand PC1 function, but that new methods and brighter tags will be required to track endogenous PC1.

Changes in growth and reproductive phenology of Allagoptera arenaria (Arecaceae) under climate change scenarios.

Climate change has led to shifts in phenology in many species distributed widely across taxonomic groups. It is, however, unclear how we should interpret these shifts without some sort of a yardstick. We assessed climate change effects on Allagoptera arenaria, a acaulescent palm, using open top chambers (OTCs) and rain gutters in the field to mimic expected temperature and rainfall changes in this area. In a coastal environment (restinga), using open top chambers (OTCs) and rain gutters in the field to mimic expected temperature and rainfall changes in this area, 40 A. arenaria individuals were selected and randomly allocated to four treatments: control (C), 25% rainfall increase (P), 2 °C temperature increase (T), and 2 °C temperature plus 25% rainfall increase (TP). For 2 years, every two weeks, we measured changes in growth and reproduction phenology to assess whether this species altered allocation patterns in response to new environmental conditions. Increases in aboveground biomass were higher in the TP than in the T treatment, which in turn had more reproductive cycles throughout the experimental period. We conclude that temperature increases may shorten the reproductive cycle of A. arenaria.

Mechanisms of Cyst Development in Polycystic Kidney Disease.

Autosomal dominant polycystic kidney disease is the most common inherited cause of end-stage kidney disease worldwide. Most cases result from mutation of either of 2 genes, PKD1 and PKD2, which encode proteins that form a probable receptor/channel complex. Studies suggest that a loss of function of the complex below an indeterminate threshold triggers cyst initiation, which ultimately results in dysregulation of multiple metabolic processes and downstream pathways and subsequent cyst growth. Noncell autonomous factors may also promote cyst growth. In this report, we focus primarily on the process of early cyst formation and factors that contribute to its variability with brief consideration of how new studies suggest this process may be reversible.

Comparative Effectiveness of COVID-19 Vaccines in Preventing Infections and Disease Progression from SARS-CoV-2 Omicron BA.5 and BA.2, Portugal.

We estimated comparative primary and booster vaccine effectiveness (VE) of SARS-CoV-2 Omicron BA.5 and BA.2 lineages against infection and disease progression. During April-June 2022, we implemented a case-case and cohort study and classified lineages using whole-genome sequencing or spike gene target failure. For the case-case study, we estimated the adjusted odds ratios (aORs) of vaccination using a logistic regression. For the cohort study, we estimated VE against disease progression using a penalized logistic regression. We observed no reduced VE for primary (aOR 1.07 [95% CI 0.93-1.23]) or booster (aOR 0.96 [95% CI 0.84-1.09]) vaccination against BA.5 infection. Among BA.5 case-patients, booster VE against progression to hospitalization was lower than that among BA.2 case-patients (VE 77% [95% CI 49%-90%] vs. VE 93% [95% CI 86%-97%]). Although booster vaccination is less effective against BA.5 than against BA.2, it offers substantial protection against progression from BA.5 infection to severe disease.

Effect of the gain rate during growing phase on the performance and characteristics of carcass and meat of super precocious steers.

The aim was to evaluate the effect of different rates of weight gain during the backgrounding on animal performance and carcass and meat characteristics of steers finished in feedlots. Thirty-six Angus steers, 12 ± 2 months of age, were backgrounded during 91 days on Aruana grass pasture (Panicum maximum cv. Aruana) managed under different stocking rates to achieve three different weight gains: HIGH ADG (average daily gain)-no feed restriction (ADG = 0.846 kg); MEDIUM ADG-moderate feed restriction (ADG = 0.456 kg); and LOW ADG-high feed restriction (ADG = 0.154 kg). To ensure the difference in ADG, we offered 0.7% live weight of concentrate feed in the HIGH treatment and a better forage supply in the MEDIUM treatment. After the backgrounding, the animals were finished in feedlot. There was no effect of the previous performance on the animals' performance in the feedlot. The LOW presented higher weight (218.9 vs. 207.9 kg) and hot (54.0% vs. 51.3%) and cold (53.5% vs. 50.5%) carcass yield than the MEDIUM, besides presenting meat with less cooking losses (15.0% vs. 18.2%), marbling (7 vs. 4.0 points), palatability (7.1 vs. 6.3 points), juiciness (7.2 vs. 6.4 points), tenderness (7.1 vs. 6.3 points), and lower shear force (5.78 vs. 8.75 kg) than HIGH. LOW ADG steers stay longer in the finishing phase but presented in general better quality carcass and meat than those with MEDIUM or HIGH during the backgrounding.

Ts17, a Tityus serrulatus ß-toxin structurally related to α-scorpion toxins.

Ts17 was purified from the venom of the scorpion Tityus serrulatus, the most dangerous scorpion species in Brazil. The activity on Nav1.1-Nav1.7 channels was electrophysiologically characterized by patch-clamp technique. Ts17 amino acid sequence indicated high similarity to alpha-scorpion toxins; however, it presented beta-toxin activity, altering the kinetics of the Na+-channels. The most affected subtypes during activation (with and without prepulse) and inactivation phases were Nav1.2 and Nav1.5, respectively. For recovery from inactivation, the most affected voltage-gated sodium channel was Nav1.5. Circular dichroism spectra showed that Ts17 presents mainly ß-sheet and unordered structures at all analyzed pHs, and the maximum value of α-helix was found at pH 4.0 (13.3 %). Based on the results, Ts17 might be used as a template to develop a new cardiac drug. Key contribution Purification of Ts17 from Tityus serrulatus, electrophysiological characterization of Ts17 on voltage-gated sodium channel subtypes, ß-toxin classification.

Invasive alien acacias rapidly stock carbon, but threaten biodiversity recovery in young second-growth forests.

Under the UN-Decade of Ecosystem Restoration and Bonn Challenge, second-growth forest is promoted as a global solution to climate change, degradation and associated losses of biodiversity and ecosystem services. Second growth is often invaded by alien tree species and understanding how this impacts carbon stock and biodiversity recovery is key for restoration planning. We assessed carbon stock and tree diversity recovery in second growth invaded by two Acacia species and non-invaded second growth, with associated edge effects, in the Brazilian Atlantic Forest. Carbon stock recovery in non-invaded forests was threefold lower than in invaded forests. Increasingly isolated, fragmented and deforested areas had low carbon stocks when non-invaded, whereas the opposite was true when invaded. Non-invaded forests recovered threefold to sixfold higher taxonomic, phylogenetic and functional diversity than invaded forest. Higher species turnover and lower nestedness in non-invaded than invaded forests underpinned higher abundance of threatened and endemic species in non-invaded forest. Non-invaded forests presented positive relationships between carbon and biodiversity, whereas in the invaded forests we did not detect any relationship, indicating that more carbon does not equal more biodiversity in landscapes with high vulnerability to invasive acacias. To deliver on combined climate change and biodiversity goals, restoration planning and management must consider biological invasion risk. This article is part of the theme issue 'Understanding forest landscape restoration: reinforcing scientific foundations for the UN Decade on Ecosystem Restoration'.

Analysis of ESAFORM 2021 cup drawing benchmark of an Al alloy, critical factors for accuracy and efficiency of FE simulations.

This article details the ESAFORM Benchmark 2021. The deep drawing cup of a 1 mm thick, AA 6016-T4 sheet with a strong cube texture was simulated by 11 teams relying on phenomenological or crystal plasticity approaches, using commercial or self-developed Finite Element (FE) codes, with solid, continuum or classical shell elements and different contact models. The material characterization (tensile tests, biaxial tensile tests, monotonic and reverse shear tests, EBSD measurements) and the cup forming steps were performed with care (redundancy of measurements). The Benchmark organizers identified some constitutive laws but each team could perform its own identification. The methodology to reach material data is systematically described as well as the final data set. The ability of the constitutive law and of the FE model to predict Lankford and yield stress in different directions is verified. Then, the simulation results such as the earing (number and average height and amplitude), the punch force evolution and thickness in the cup wall are evaluated and analysed. The CPU time, the manpower for each step as well as the required tests versus the final prediction accuracy of more than 20 FE simulations are commented. The article aims to guide students and engineers in their choice of a constitutive law (yield locus, hardening law or plasticity approach) and data set used in the identification, without neglecting the other FE features, such as software, explicit or implicit strategy, element type and contact model.

Tracking SARS-CoV-2 lineage B.1.1.7 dissemination: insights from nationwide spike gene target failure (SGTF) and spike gene late detection (SGTL) data, Portugal, week 49 2020 to week 3 2021.

We show that the SARS-CoV-2 B.1.1.7 lineage is highly disseminated in Portugal, with the odds of B.1.1.7 proportion increasing at an estimated 89% (95% confidence interval: 83-95%) per week until week 3 2021. RT-PCR spike gene target late detection (SGTL) can constitute a useful surrogate to track B.1.1.7 spread, besides the spike gene target failure (SGTF) proxy. SGTL/SGTF samples were associated with statistically significant higher viral loads, but not with substantial shift in age distribution compared to non-SGTF/SGTL cases.

Pkd1 Mutation Has No Apparent Effects on Peroxisome Structure or Lipid Metabolism.

Background: Multiple studies of tissue and cell samples from patients and preclinical models of autosomal dominant polycystic kidney disease report abnormal mitochondrial function and morphology and suggest metabolic reprogramming is an intrinsic feature of this disease. Peroxisomes interact with mitochondria physically and functionally, and congenital peroxisome biogenesis disorders can cause various phenotypes, including mitochondrial defects, metabolic abnormalities, and renal cysts. We hypothesized that a peroxisomal defect might contribute to the metabolic and mitochondrial impairments observed in autosomal dominant polycystic kidney disease. Methods: Using control and Pkd1-/- kidney epithelial cells, we investigated peroxisome abundance, biogenesis, and morphology by immunoblotting, immunofluorescence, and live cell imaging of peroxisome-related proteins and assayed peroxisomal specific ß-oxidation. We further analyzed fatty acid composition by mass spectrometry in kidneys of Pkd1fl/fl;Ksp-Cre mice. We also evaluated peroxisome lipid metabolism in published metabolomics datasets of Pkd1 mutant cells and kidneys. Lastly, we investigated if the C terminus or full-length polycystin-1 colocalize with peroxisome markers by imaging studies. Results: Peroxisome abundance, morphology, and peroxisome-related protein expression in Pkd1-/- cells were normal, suggesting preserved peroxisome biogenesis. Peroxisomal ß-oxidation was not impaired in Pkd1-/- cells, and there was no obvious accumulation of very-long-chain fatty acids in kidneys of mutant mice. Reanalysis of published datasets provide little evidence of peroxisomal abnormalities in independent sets of Pkd1 mutant cells and cystic kidneys, and provide further evidence of mitochondrial fatty acid oxidation defects. Imaging studies with either full-length polycystin-1 or its C terminus, a fragment previously shown to go to the mitochondria, showed minimal colocalization with peroxisome markers restricted to putative mitochondrion-peroxisome contact sites. Conclusions: Our studies showed that loss of Pkd1 does not disrupt peroxisome biogenesis nor peroxisome-dependent fatty acid metabolism.

Leaching with mixed organic acids and sulfuric acid to recover cobalt and lithium from lithium ion batteries.

Li-ion batteries (LIBs) should be recycled because of the environmental reasons and this type of waste represents an important secondary source of metals. This work aimed to evaluate the recovery of Co and Li from LIBs by hydrometallurgy. The efficiency of different leachants was tested: H2SO4 (2 M), fermentation effluent with supplementation of organic acids (lactic, acetic, butyric and propionic acids) (3.4 M) and a combination of fermentation effluent (0.75 M) and H2SO4 (1.25 M). In addition, the effect of H2O2, glucose P.A., lactose P.A. and from milk whey permeate (MWP) as reducing agent was tested. The leaching solution composed of H2SO4 and fermentation effluent showed high potential of metals recovery in addition to being an alternative of reducing the volume of inorganic acid and the cost by using a fermentation effluent since its use may be integrated with a waste treatment process. Based on Central Composite Designs, optimum conditions of leaching were established, as temperature of 86°C, solid-liquid ratio of 18.5 g/L, leaching time 2.5 h, agitation of 300 rpm and concentration of 0.09 M of lactose from MWP and recovery level achieved was 93.35% of Co and 90.50% of Li. In order to evaluate the influence of each organic acid present on the fermentation effluent, testes were carried out using pure organic acid with H2SO4 (0.75 M:1.25 M) or isolated (3.4 M) and inferior recoveries were detected proving that mixture of organic acids and further compounds as phenolic groups characteristic of fermentation effluent improves the leaching process.

A força de atração a ser-na-massa: comentários ao artigo "O assintótico", de Pierre Fédida / The force of attraction to populism: comments on the article "The Asymptote" by Pierre Fédida / La fuerza de atracción a ser de la masa: comentarios sobre el artículo "Lo assintótico" de Pierre Fédida / La force d'attraction de l'être dans les masses: commentaires à l'article « L'asymptotique ¼ de Pierre Fédida

RESUMO Este texto é um breve comentário ao artigo "O assintótico", de Fédida, destacando aspectos da escuta psicanalítica e várias facetas que decorrem das peculiaridades de seu objeto, um inconsciente, fragmentário, apreendido apenas nas brechas de uma formação como o eu que tende à unificação totalizante, sintética. A análise, voltada para particularidades cruciais de cada um, se faz, pois, à contracorrente de aspirações conformistas e massificadoras inerentes ao narcisismo.

ABSTRACT This work is a brief comment to the article written by Fédida, The Asymptote, highlighting aspects of the psychoanalytic listening and the several facets derived from peculiarities of its object, the unconscious, fragmentary, captured as the I is formed, tending to total and synthetic unification. The analysis directed to essential particularities of each individual, is done contrary to conformists and populist desires associated to narcissism.

RESUMEN Ese texto es un breve comentario sobre el artículo de Fédida, "O assintótico", resaltando aspectos de la escucha psicoanalítica y varias facetas derivadas de las peculiaridades de su objeto, un inconsciente, fragmentario, capturado solamente en las brechas de una formación como el Yo que tiende a la unificación totalizadora, sintética. Por lo tanto, el análisis, orientado a las particularidades cruciales de cada uno, se vuelve en contra de las aspiraciones conformistas y masificadoras inherentes al narcisismo.

RÉSUMÉ Ce texte est un bref commentaire à l'article de Fédida, « L'asymptotique ¼, soulignant des aspects d'écoute psychanalytique et plusieurs facettes qui découlent des particularités de son objet, un inconscient fragmentaire, qui n'est appréhendé que dans les brèches d'une formation comme le Moi, qui tend à l'unification totalisante, synthétique. L'analyse, tournée vers les particularités cruciales de chacun, se fait donc à contrecourant d'aspirations conformistes et capables de massifier, inhérentes au narcissisme.

Epilepsy-Related Voltage-Gated Sodium Channelopathies: A Review.

Epilepsy is a disease characterized by abnormal brain activity and a predisposition to generate epileptic seizures, leading to neurobiological, cognitive, psychological, social, and economic impacts for the patient. There are several known causes for epilepsy; one of them is the malfunction of ion channels, resulting from mutations. Voltage-gated sodium channels (NaV) play an essential role in the generation and propagation of action potential, and malfunction caused by mutations can induce irregular neuronal activity. That said, several genetic variations in NaV channels have been described and associated with epilepsy. These mutations can affect channel kinetics, modifying channel activation, inactivation, recovery from inactivation, and/or the current window. Among the NaV subtypes related to epilepsy, NaV1.1 is doubtless the most relevant, with more than 1500 mutations described. Truncation and missense mutations are the most observed alterations. In addition, several studies have already related mutated NaV channels with the electrophysiological functioning of the channel, aiming to correlate with the epilepsy phenotype. The present review provides an overview of studies on epilepsy-associated mutated human NaV1.1, NaV1.2, NaV1.3, NaV1.6, and NaV1.7.

Pathway identification through transcriptome analysis.

Systems-based, agnostic approaches focusing on transcriptomics data have been employed to understand the pathogenesis of polycystic kidney diseases (PKD). While multiple signaling pathways, including Wnt, mTOR and G-protein-coupled receptors, have been implicated in late stages of disease, there were few insights into the transcriptional cascade immediately downstream of Pkd1 inactivation. One of the consistent findings has been transcriptional evidence of dysregulated metabolic and cytoskeleton remodeling pathways. Recent technical developments, including bulk and single-cell RNA sequencing technologies and spatial transcriptomics, offer new angles to investigate PKD. In this article, we review what has been learned based on transcriptional approaches and consider future opportunities.

Secondary forest fragments offer important carbon and biodiversity cobenefits.

Tropical forests store large amounts of carbon and high biodiversity, but are being degraded at alarming rates. The emerging global Forest and Landscape Restoration (FLR) agenda seeks to limit global climate change by removing carbon dioxide from the atmosphere through the growth of trees. In doing so, it may also protect biodiversity as a free cobenefit, which is vital given the massive shortfall in funding for biodiversity conservation. We investigated whether natural forest regeneration on abandoned pastureland offers such cobenefits, focusing for the first time on the recovery of taxonomic diversity (TD), phylogenetic diversity (PD) and functional diversity (FD) of trees, including the recovery of threatened and endemic species richness, within isolated secondary forest (SF) fragments. We focused on the globally threatened Brazilian Atlantic Forest, where commitments have been made to restore 1 million hectares under FLR. Three decades after land abandonment, regenerating forests had recovered ~20% (72 Mg/ha) of the above-ground carbon stocks of a primary forest (PF), with cattle pasture containing just 3% of stocks relative to PFs. Over this period, SF recovered ~76% of TD, 84% of PD and 96% of FD found within PFs. In addition, SFs had on average recovered 65% of threatened and ~30% of endemic species richness of primary Atlantic forest. Finally, we find positive relationships between carbon stock and tree diversity recovery. Our results emphasize that SF fragments offer cobenefits under FLR and other carbon-based payments for ecosystem service schemes (e.g. carbon enhancements under REDD+). They also indicate that even isolated patches of SF could help to mitigate climate change and the biodiversity extinction crisis by recovering species of high conservation concern and improving landscape connectivity.

Intramuscular fatty acid composition of steers finished on oat pastures mixture with vetch or cornmeal supplementation / Composição de ácidos graxos da gordura intramuscular de novilhos terminados em pastagem de aveia consorciada com ervilhaca ou suplementados com grão de milho moído

ABSTRACT: The effects of energetic supplementation and mixture vetch (Vica sativa L.) were studied to evaluate the intramuscular fatty acid profile of steers finished on oat (Avena sativa L.) pastures. Eighteen 21-month-old crossbred steers were evaluated. The experiment was carried out in three treatments and six repetitions: oat pasture (OA), oat pasture + vetch (OA + VET), and oat pasture + supplementation (OA + SUP). Supplementation comprised the addition of corn meal at a daily dose of 1% of the animals' body weight. Steers receiving supplementation had higher lipid content in Longissimus lumborum than did those fed with OA + VET (1.25 vs 1.02%), whereas those fed with OA (1.15%) showed intermediate values. Conjugated linoleic acid levels were higher in steers fed OA (0.57%) and OA + SUP (0.59%), whereas the highest amount of omega-3 fatty acids was observed in animals fed OA + VET (3.32%). Pasture-finishing without supplementation resulted in a lower ratio of n-6:n-3 (3.14). Steers finished in oat mixture with vetch showed healthier intramuscular fat than did those finished with cornmeal supplementation; importantly, the higher the amount of PUFAs, the better n-6:n-3 ratio.

RESUMO: Os efeitos da suplementação energética e da ervilhaca (Vica sativa L.) foram estudados para avaliar o perfil de ácidos graxos intramusculares de novilhos terminados em pastagens de aveia (Avena sativa L.). Foram avaliados 18 novilhos mestiços com 21 meses de idade. O experimento foi realizado em três tratamentos e seis repetições: pastagem de aveia (OA), pastagem de aveia + ervilhaca (OA + VET) e pastagem de aveia + suplementação (OA + SUP). A suplementação compreendeu a adição de fubá na dose diária de 1% do peso corporal dos animais. Os novilhos que receberam suplementação apresentaram maior teor lipídico no Longissimus lombar do que aqueles alimentados com OA + VET (1,25 contra 1,02%), enquanto aqueles alimentados com AO (1,15%) apresentaram valores intermediários. Os níveis de ácido linoléico conjugado foram maiores em novilhos alimentados com AO (0,57%) e OA + SUP (0,59%), enquanto a maior quantidade de ácidos graxos ômega-3 foi observada em animais alimentados com OA + VET (3,32%). Animais terminados em pastagens sem suplementação apresentaram menor proporção de n-6: n-3 (3,14). Novilhos terminados em pastagem de aveia com ervilhaca apresentaram gordura intramuscular mais saudável do que aqueles terminados com suplementação de milho moído, uma vez que apresentaram maior quantidade de PUFAs e melhor a relação n-6: n-3.

Natural Occurrence in Venomous Arthropods of Antimicrobial Peptides Active against Protozoan Parasites.

Arthropoda is a phylum of invertebrates that has undergone remarkable evolutionary radiation, with a wide range of venomous animals. Arthropod venom is a complex mixture of molecules and a source of new compounds, including antimicrobial peptides (AMPs). Most AMPs affect membrane integrity and produce lethal pores in microorganisms, including protozoan pathogens, whereas others act on internal targets or by modulation of the host immune system. Protozoan parasites cause some serious life-threatening diseases among millions of people worldwide, mostly affecting the poorest in developing tropical regions. Humans can be infected with protozoan parasites belonging to the genera Trypanosoma, Leishmania, Plasmodium, and Toxoplasma, responsible for Chagas disease, human African trypanosomiasis, leishmaniasis, malaria, and toxoplasmosis. There is not yet any cure or vaccine for these illnesses, and the current antiprotozoal chemotherapeutic compounds are inefficient and toxic and have been in clinical use for decades, which increases drug resistance. In this review, we will present an overview of AMPs, the diverse modes of action of AMPs on protozoan targets, and the prospection of novel AMPs isolated from venomous arthropods with the potential to become novel clinical agents to treat protozoan-borne diseases.

The pathobiology of polycystic kidney disease from a metabolic viewpoint.

Autosomal dominant polycystic kidney disease (ADPKD) affects an estimated 1 in 1,000 people and slowly progresses to end-stage renal disease (ESRD) in about half of these individuals. Tolvaptan, a vasopressin 2 receptor blocker, has been approved by regulatory authorities in many countries as a therapy to slow cyst growth, but additional treatments that target dysregulated signalling pathways in cystic kidney and liver are needed. Metabolic reprogramming is a prominent feature of cystic cells and a potentially important contributor to the pathophysiology of ADPKD. A number of pathways previously implicated in the pathogenesis of the disease, such as dysregulated mTOR and primary ciliary signalling, have roles in metabolic regulation and may exert their effects through this mechanism. Some of these pathways are amenable to manipulation through dietary modifications or drug therapies. Studies suggest that polycystin-1 and polycystin-2, which are encoded by PKD1 and PKD2, respectively (the genes that are mutated in >99% of patients with ADPKD), may in part affect cellular metabolism through direct effects on mitochondrial function. Mitochondrial dysfunction could alter the redox state and cellular levels of acetyl-CoA, resulting in altered histone acetylation, gene expression, cytoskeletal architecture and response to cellular stress, and in an immunological response that further promotes cyst growth and fibrosis.