Analysis of learning curves in predictive modeling using exponential curve fitting with an asymptotic approach.

The existence of large volumes of data has considerably alleviated concerns regarding the availability of sufficient data instances for machine learning experiments. Nevertheless, in certain contexts, addressing limited data availability may demand distinct strategies and efforts. Analyzing COVID-19 predictions at pandemic beginning emerged a question: how much data is needed to make reliable predictions? When does the volume of data provide a better understanding of the disease's evolution and, in turn, offer reliable forecasts? Given these questions, the objective of this study is to analyze learning curves obtained from predicting the incidence of COVID-19 in Brazilian States using ARIMA models with limited available data. To fulfill the objective, a retrospective exploration of COVID-19 incidence across the Brazilian States was performed. After the data acquisition and modeling, the model errors were assessed by employing a learning curve analysis. The asymptotic exponential curve fitting enabled the evaluation of the errors in different points, reflecting the increased available data over time. For a comprehensive understanding of the results at distinct stages of the time evolution, the average derivative of the curves and the equilibrium points were calculated, aimed to identify the convergence of the ARIMA models to a stable pattern. We observed differences in average derivatives and equilibrium values among the multiple samples. While both metrics ultimately confirmed the convergence to stability, the equilibrium points were more sensitive to changes in the models' accuracy and provided a better indication of the learning progress. The proposed method for constructing learning curves enabled consistent monitoring of prediction results, providing evidence-based understandings required for informed decision-making.

The Cardiometabolic Impact of Rebaudioside A Exposure during the Reproductive Stage.

The consumption of non-sugar sweeteners (NSS) has increased during pregnancy. The European Food Safety Agency suggested that steviol glycosides, such as Rebaudioside A (RebA), the major sweetener component of stevia, are safe for humans up to a dose of 4 mg/kg body weight/day. However, the World Health Organization recommended in 2023 the restraint of using NSS, including stevia, at any life stage, highlighting the need to study NSS safety in early periods of development. We aimed to study the mitochondrial and cardiometabolic effects of long-term RebA consumption during the reproductive stage of the life cycle. Female rats were exposed to RebA (4 mg steviol equivalents/kg body weight/day) in the drinking water from 4 weeks before mating until weaning. Morphometry, food and water consumption, glucose and lipid homeostasis, heart structure, function, and mitochondrial function were assessed. RebA showed an atrophic effect in the heart, decreasing cardiomyocyte cross-sectional area and myocardial fibrosis without repercussions on cardiac function. Mitochondrial and myofilamentary functions were not altered. Glucose tolerance and insulin sensitivity were not affected, but fasting glycemia and total plasma cholesterol decreased. This work suggests that this RebA dose is safe for female consumption during the reproductive stage, from a cardiometabolic perspective. However, studies on the effects of RebA exposure on the offspring are mandatory.

Acute and chronic effects of levosimendan in the ZSF1 obese rat model of heart failure with preserved ejection fraction.

Heart failure with preserved ejection fraction (HFpEF) is a syndrome characterized by impaired cardiovascular reserve in which therapeutic options are scarce. Our aim was to evaluate the inodilator levosimendan in the ZSF1 obese rat model of HFpEF. Twenty-week-old male Wistar-Kyoto (WKY), ZSF1 lean (ZSF1 Ln) and ZSF1 obese rats chronically treated for 6-weeks with either levosimendan (1 mg/kg/day, ZSF1 Ob + Levo) or vehicle (ZSF1 Ob + Veh) underwent peak-effort testing, pressure-volume (PV) haemodynamic evaluation and echocardiography (n = 7 each). Samples were collected for histology and western blotting. In obese rats, skinned and intact left ventricular (LV) cardiomyocytes underwent in vitro functional evaluation. Seven additional ZSF1 obese rats underwent PV evaluation to assess acute levosimendan effects (10 µg/kg + 0.1 µg/kg/min). ZSF1 Ob + Veh presented all hallmarks of HFpEF, namely effort intolerance, elevated end-diastolic pressures and reduced diastolic compliance as well as increased LV mass and left atrial area, cardiomyocyte hypertrophy and increased interstitial fibrosis. Levosimendan decreased systemic arterial pressures, raised cardiac index, and enhanced LV relaxation and diastolic compliance in both acute and chronic experiments. ZSF1 Ob + Levo showed pronounced attenuation of hypertrophy and interstitial fibrosis alongside increased effort tolerance (endured workload raised 38 %) and maximum O2 consumption. Skinned cardiomyocytes from ZSF 1 Ob + Levo showed a downward shift in sarcomere length-passive tension relationship and intact cardiomyocytes showed decreased diastolic Ca2+ levels and enhanced Ca2+ sensitivity. On molecular grounds, levosimendan enhanced phosphorylation of phospholamban and mammalian target of rapamycin. The observed effects encourage future clinical trials with levosimendan in a broad population of HFpEF patients.

Non-News Websites Expose People to More Political Content Than News Websites: Evidence from Browsing Data in Three Countries.

Most scholars focus on the prevalence and democratic effects of (partisan) news exposure. This focus misses large parts of online activities of a majority of politically disinterested citizens. Although political content also appears outside of news outlets and may profoundly shape public opinion, its prevalence and effects are under-studied at scale. This project combines three-wave panel survey data from three countries (total N = 7,266) with online behavioral data from the same participants (over 106M visits). We create a multi-lingual classifier to identify political content both in news and outside (e.g. in shopping or entertainment sites). We find that news consumption is infrequent: just 3.4% of participants' online browsing comprised visits to news sites. Only between 14% (NL) and 36% (US) of these visits were to news about politics. The overwhelming majority of participants' visits were to non-news sites. Although only 1.6\% of those visits related to politics, in absolute terms, citizens encounter politics more frequently outside of news than within news. Out of every 10 visits to political content, 3.4 come from news and 6.6 from non-news sites. Furthermore, exposure to political content outside news domains had the same - and in some cases stronger - associations with key democratic attitudes and behaviors as news exposure. These findings offer a comprehensive analysis of the online political (not solely news) ecosystem and demonstrate the importance of assessing the prevalence and effects of political content in non-news sources.

Synthesis of nanoporous carbonaceous materials at lower temperatures.

Nanoporous carbonaceous materials are ideal ingredients in various industrial products due to their large specific surface area. They are typically prepared by post-synthesis activation and templating methods. Both methods require the input of large amounts of energy to sustain thermal treatment at high temperatures (typically >600°C), which is clearly in violation of the green-chemistry principles. To avoid this issue, other strategies have been developed for the synthesis of carbonaceous materials at lower temperatures (<600°C). This mini review is focused on three strategies suitable for processing carbons at lower temperatures, namely, hydrothermal carbonization, in situ hard templating method, and mechanically induced self-sustaining reaction. Typical procedures of these strategies are demonstrated by using recently reported examples. At the end, some problems associated with the strategies and potential solutions are discussed.

Effects of Triiodothyronine Treatment in an Animal Model of Heart Failure with Preserved Ejection Fraction.

Background: Low levels of triiodothyronine (T3) are common in patients with heart failure (HF). Our aim was to evaluate the effects of supplementation with low and replacement doses of T3 in an animal model of HF with preserved ejection fraction (HFpEF). Methods: We evaluated four groups: ZSF1 Lean (n = 8, Lean-Ctrl), ZSF1 Obese (rat model of metabolic-induced HFpEF, n = 13, HFpEF), ZSF1 Obese treated with a replacement dose of T3 (n = 8, HFpEF-T3high), and ZSF1 Obese treated with a low-dose of T3 (n = 8, HFpEF-T3low). T3 was administered in drinking water from weeks 13 to 24. The animals underwent anthropometric and metabolic assessments, echocardiography, and peak effort testing with maximum O2 consumption (VO2max) determination at 22 weeks, and a terminal hemodynamic evaluation at 24 weeks. Afterwhile myocardial samples were collected for single cardiomyocyte evaluation and molecular studies. Results: HFpEF animals showed lower serum and myocardial thyroid hormone levels than Lean-Ctrl. Treatment with T3 did not normalize serum T3 levels, but increased myocardial T3 levels to normal levels in the HFpEF-T3high group. Body weight was significantly decreased in both the T3-treated groups, comparing with HFpEF. An improvement in glucose metabolism was observed only in HFpEF-T3high. Both the treated groups had improved diastolic and systolic function in vivo, as well as improved Ca2+ transients and sarcomere shortening and relaxation in vitro. Comparing with HFpEF animals, HFpEF-T3high had increased heart rate and a higher rate of premature ventricular contractions. Animals treated with T3 had higher myocardial expression of calcium transporter ryanodine receptor 2 (RYR2) and α-myosin heavy chain (MHC), with a lower expression of ß-MHC. VO2max was not influenced by treatment with T3. Myocardial fibrosis was reduced in both the treated groups. Three animals died in the HFpEF-T3high group. Conclusions: Treatment with T3 was shown to improve metabolic profile, myocardial calcium handling, and cardiac function. While the low dose was well-tolerated and safe, the replacement dose was associated with increased heart rate, and increased risk of arrhythmias and sudden death. Modulation of thyroid hormones may be a potential therapeutic target in HFpEF; however, it is important to take into account the narrow therapeutic window of T3 in this condition.

Combining Zeolites with Early-Maturing Annual Legume Cover Crops in Rainfed Orchards: Effects on Yield, Fatty Acid Composition and Polyphenolic Profile of Olives and Olive Oil.

Under climate change threats, there is a growing need to adapt the conventional agronomic practices used in rainfed olive orchards by sustainable practices, in order to ensure adequate crop yield and olive oil quality and to preserve soil health. Therefore, for two years, the effects of conventional tillage practice (T) and two sustainable soil management strategies, a leguminous cover crop (LC) and its combination with natural zeolites (ZL), on the yield, fatty acid composition, polyphenolic profile and quality indices of olive fruits and oil were evaluated. Crop yield was significantly increased by LC and ZL in the first year. Although in the second year no significant differences were verified, the cumulative yield increased significantly by 31.6% and 35.5% in LC and ZL trees, respectively. LC enhanced the moisture and size of olives, while ZL increased, in general, the concentrations of oleuropein, verbascoside, caffeic acid and epicatechin, as well the oleic/linoleic ratio in fruits and the levels of 3,4-dihydroxyphenylglycol, tyrosol, verbascoside and caffeic acid in olive oil. Despite the higher concentration of total phenols in the fruits and oil from T trees in the warmer and dryer year, the quality of the oil decreased, mainly when compared with ZL, as evidenced by the peroxide value and K232 and K270 coefficients. In short, both sustainable soil management strategies appear to be promising practices to implement in olive orchards under rainfed conditions, but the innovative strategy of combining zeolites with legume cover crops, first reported in the present study, confers advantages from a nutritional and technological point of view. Nevertheless, studies subjected to the long-term use of these practices should be conducted to ensure the sustainability of the crop yield and olive oil quality.

Diet-Induced Microbiome's Impact on Heart Failure: A Double-Edged Sword.

Heart failure (HF) is a debilitating disease with a significant clinical and economic impact worldwide. Multiple factors seem to increase the risk of developing HF, such as hypertension, obesity and diabetes. Since chronic inflammation plays a significant role in HF pathophysiology and gut dysbiosis is associated with low-grade chronic inflammation, the risk of cardiovascular diseases is likely modulated by the gut microbiome (GM). Considerable progress has been made in HF management. However, there is a need to find new strategies to reduce mortality and increase the quality of life, mainly of HFpEF patients, since its prevalence continues to rise. Recent studies validate that lifestyle changes, such as diet modulation, represent a potential therapeutic approach to improve several cardiometabolic diseases, although their effects on the GM and its indirect cardiac impact still warrant further research. Hence, in this paper, we aim to clarify the link between HF and the human microbiome.

Olive Yield and Physicochemical Properties of Olives and Oil in Response to Nutrient Application under Rainfed Conditions.

The effects of mineral fertilizers on the physicochemical properties of olives and oil under rainfed conditions is scarce. In this three-year study, the results of a nitrogen (N), phosphorus (P), potassium (K) and boron (B) fertilization trial carried out in a young rainfed olive grove and arranged as a nutrient omission trial are reported. The control consisted of the application of N, P, K and B (NPKB) and four other treatments corresponded to the removal of one of them (N0, P0, K0 and B0). Olive yield and several variables associated with the physicochemical properties of olives and oil were evaluated. The NPKB treatment increased olive yield compared to the treatment that did not receive N (N0). Although dependent on the climate conditions of the crop season, the NPKB treatment increased fruit weight and the pulp/pit ratio and its fruits tended to accumulate more oil than K0. However, the phenolics concentrations on fruits and oil tended to be lower. All olive oil samples were classified in the "extra virgin" category and all showed a decrease in its stability between 3 and 15 months of storage, regardless of treatment, especially in N0, P0 and B0 treatments. The results of the sensorial analysis indicate that all the oils fell into the medium fruitiness and greenly-fruity category. Only in P0 and B0 were defects detected, namely muddy sediment. Thus, this study seems to indicate the importance of N application, but also a balanced nutrient application and that further studies are needed, given the difficulty in finding clear trends in the response of measured variables to fertilizer treatments.

Physical capacity, body composition and immune-inflammatory markers are associated with risk of cardiovascular disease in sedentary people living with HIV.

BACKGROUND: People living with HIV (PLHIV) are experiencing a high level of non-HIV-related disease mainly related to cardiovascular system however the factors associated with this scenario are widely discussed and indefinite so far. The purpose was to investigate the association of physical capacities, body composition, and immune-inflammatory markers with the Framingham risk score (FRS) in PLHIV. METHODS: Volunteers were recruited and data on age, the time of HIV infection diagnosis and the time of HAART use were collected. The volunteers performed upper limb strength (ULS), lower limb strength (LLS) and cardiorespiratory fitness (VO2peak) tests, body composition evaluation, and blood collection to immune-inflammatory marker markers and the FRS was calculated. Partial age- and sex-adjusted correlation was used to verify associations between factors and multiple linear regression analysis was performed to identify an independent association of FRS and the variables. RESULTS: 113 PLHIV met the eligible criteria and were observed a negative correlation between CD4+ (r=-0.615), time of HIV infection diagnosis (r=-0.237), time of HAART use (r=-0.239), ULS (r=-0.274), LLS (r=-0.213), and VO2peak (r=-0.207) with FRS. There was a positive correlation between fibrinogen and FRS (r=0.363; P<0.001). The TCD4+ count cells were significantly associated with FRS (P=0.001). CONCLUSIONS: Environmental and biological HIV-related factors are inversely associated with FRS while immune-inflammation markers have a positive association.

Boas práticas em saúde integral para travestis, transexuais e outras variabilidades de gênero/gêneros diversos: experiência do Ambulatório de Saúde Integral para Travestis e Transexuais (ASITT) do Centro de Referência e Treinamento DST/Aids-SP, Secretaria de Estado da Saúde de São Paulo / Good practices in integral health for travestis, transsexuals and other variabilities of gender/different genders: experience of the Health Outpatient Clinic for Travestis and Transsexuals (ASITT) of the STD/AIDS Reference and Training Center-SP, State Department of Health of Sao Paulo

O Centro de Referência e Treinamento DST/Aids-SP da Secretaria de Estado da Saúde de São Paulo, inaugurou, em junho de 2009, em suas dependências, o primeiro ambulatório voltado exclusivamente à saúde integral de travestis e transexuais do país, com ênfase no processo transexualizador do Sistema Único de Saúde (SUS) e nas necessidades dessa população. Este artigo tem por objetivo apresentar as modalidades assistenciais ofertadas no serviço, as demandas da população atendida no ambulatório, suas vulnerabilidades, fatores de risco e resiliência a que está submetida, assim como as considerações e os desafios no campo da política pública voltada à população trans para promover seu acesso à atenção em saúde de forma acolhedora, livre de preconceito, julgamentos morais e práticas discriminatórias.

A implementação da linha de cuidado para travestis e transexuais e outras variabilidades de gênero no estado de São Paulo / The implementation of the line of care for travestis and transsexuals and other gender variability in the state of Sao Paulo

Este texto tem por finalidade apresentar a linha de cuidado (LC) planejada e articulada com a implementação de novos serviços, integrada a um conjunto de práticas mais abrangentes de assistência em saúde e aos demais setores responsáveis por políticas públicas que tenham como objetivo atender travestis e transexuais e outras variabilidades de gênero. Consideramos que a implementação de atenção integral à população travesti e transexual envolve um conjunto de ações e procedimentos em saúde ofertados para melhor qualificação e proposta para demandas de experiências integrais de pessoas trans e com variabilidade de gênero no campo da saúde e do Sistema Único de Saúde (SUS), incluindo modificação das características corporais, com o objetivo de adquirir e expressar características do gênero com o qual essas pessoas se identificam e ao qual declaram pertencer.

Zeolites and Biochar Modulate Olive Fruit and Oil Polyphenolic Profile.

Soil degradation processes and climate change threaten the sustainability of Mediterranean rainfed olive orchards, with repercussions on crop yield and quality of olives, olive oil and olive by-products. Using soil amendments can enhance soil fertility for sustained environmental quality and plant performance. For two years, we evaluated, under rainfed conditions, the effects of a fertilizer compound (FC) and its combination with zeolites (ZL) and biochar (BC) amendments on soil moisture, yield, fruit and oil polyphenols and quality indices. The polyphenolic composition was strongly influenced by treatments, although no effects were observed on crop yield. ZL improved soil moisture (average increase of 26.3% compared to FC), fruit fatty acid composition (increase of 12.4% in oleic/linoleic ratio in 2018) and oil quality, BC enhanced the concentrations of polyphenols with high nutritional value (average annual increase of 25.6, 84.8 and 11.6% for 3,4-dihydroxyphenylglycol, oleuropein and rutin, respectively). In contrast, olive oil from FC fruits showed the poorest quality, with oxidation and hydrolytic breakdown signals. The applied soil amendments appear to be a promising sustainable strategy to implement in olive rainfed orchards.

Endoscopic analysis of alterations in sinus floor elevation approaching the summers technique and immediate implant: a clinical trial with 10 years follow-up / Análise endoscópica das alterações na elevação do assoalho do seio maxilar com abordagem da técnica de Summer e implante imediato: um ensaio clínico com 10 anos de acompanhamento

The goal of this study was to evaluate by endoscopy, possible intercurrences during the sinus floor lifting employing the Summers technique, besides verify the implants' survival rate after 10 years. Six patients (12 sinus) were included in this study. The same surgeon performed the procedures, under local anesthesia and venous sedation in the hospital. All participants underwent sinus lift and implant placement in only one procedure, as previously planned, using endoscopic analysis (Stortz®). All cases received bovine bone graft (Bio-Oss®) before the implant placement. After 10 years, the patients were recalled for follow-up. Two intercurrences (16.66%) were detected using the endoscope, one simple rupture, and another perforation with the leaking of the graft within the sinus. Both were reverted and corrected immediately. There was one implant loss (8.33%), therefore this patient did not undergo any intercurrence in transoperative, and the membrane was elevated lesser than 5 mm. The survival rate reached was 91.66%. The osteotome technique constitutes a reliable method with a long-term of 10 years presenting a high implant survival rate, suggesting an elevation up to 5.5 mm in healthy patients. The occurrences in transoperative were only detected by the endoscopic analysis which must be stimulated to guarantee more secure visibility. Otherwise, the association the atraumatic technique and endoscope was tough, increased the costs, limiting the use routinely.

O objetivo deste estudo foi avaliar através de endoscopia as possíveis intercorrências durante levantamento de seio maxilar fechado, além de verificar a taxa de sobrevivência dos implantes após 10 anos. Seis pacientes (12 seios maxilares) foram incluídos neste estudo com idades entre 26 e 74 anos. O mesmo cirurgião realizou os procedimentos, sob anestesia local e sedação venosa em ambiente hospitalar. Todos os participantes foram submetidos à elevação do seio nasal e colocação do implante em apenas um procedimento, conforme planejado anteriormente, por acompanhamento transcirúrgico da endoscopia (Stortz®). Todos os casos receberam enxerto ósseo bovino (Bio-Oss®) antes da colocação do implante. Após 10 anos, os pacientes foram chamados para acompanhamento. Foram detectadas duas intercorrências (16,66%) com o endoscópio, uma ruptura simples e outra perfuração com extravasamento do enxerto para dentro do seio. Ambos foram revertidos e corrigidos imediatamente. Houve perda de um implante (8,33%), portanto esse paciente não apresentou intercorrência no transoperatório e a membrana estava elevada menos de 5 mm. A taxa de sobrevivência alcançada foi de 91,66%. A técnica do osteótomo constitui um método confiável em longo prazo (10 anos) apresentando uma alta taxa de sobrevivência do implante, sugerindo que uma elevação de até 5,5 mm em pacientes saudáveis é possível. Assim, verificou-se que as ocorrências no transoperatório foram detectadas apenas pela análise endoscópica que deve ser estimulada para garantir uma visibilidade mais segura. Por outro lado, a associação da técnica fechada com o endoscópio foi difícil, aumentou os custos, limitando seu uso como rotina

Evaluation of myenteric neurons in the colon of rats exposed to 2,4 dichlorophenoxyacetic acid herbicide.

The assessment of the enteric nervous system provides a better understanding of the effects that contaminants can have on the health and well-being of organisms. It has been reported that 2,4-dichlorophenoxyacetic acid (2,4-D) is a highly persistent herbicide in the environment that is responsible for neurotoxic changes in different myenteric neuronal subpopulations. The current study aimed to evaluate the effects of 2,4-D on myenteric neurons in the colon of Rattus norvegicus for the first time. A dose of 2,4-D (5 mg/kg/day) was administered to the experimental group (2,4-D) for 15 days. Then, the proximal colon was collected and submitted to Giemsa and NADPH-d histochemical techniques for the disclosure of total and nitrergic neurons. The 2,4-D group presented a higher density of total neurons (p = 0.05, t-test), which together with the maintenance of nitrergic neuronal density, may be related to the increase in the expression of the neurotransmitter acetylcholine by colocalization, responsible for stimulating the intestinal smooth muscle and increasing the chances of the expulsion of the harmful content present in the lumen. Over 15 days, the neurotoxic effects of 2,4-D in the myenteric plexus influenced an increase in the general population of myenteric neurons in the colon.

Integrated analysis of fish intestine biomarkers: Complementary tools for pollution assessment.

The gastrointestinal tract and its enteric nervous system are the first routes of food and xenobiotics uptake. Considering the importance of this organ, this study evaluated intestinal biomarkers of Sphoeroides testudineus integrating the data to generate tools for pollution assessment. The fish were collected in three sites of São Paulo Coast and their intestines were analyzed for biochemical, histology, and neuronal density and morphometry biomarkers. To evaluate the differences among the data, a PERMANOVA was applied, followed by a FA/PCA. The PERMANOVA indicated differences (P < 0.001) between the regions (RA, A1, and A2). Four factors were extracted from the FA/PCA (62% cumulative), showing that the animals from A2 presented severe alterations, mainly in intestinal morphometry and neuronal density. A1 alterations refer mainly to the increase of neuronal metabolism. Our results also evidence a gradient of environmental quality related to the protection level (AR > A1 > A2).

Active Actions in the Extraction of Urban Objects for Information Quality and Knowledge Recommendation with Machine Learning.

Due to the increasing urban development, it has become important for municipalities to permanently understand land use and ecological processes, and make cities smart and sustainable by implementing technological tools for land monitoring. An important problem is the absence of technologies that certify the quality of information for the creation of strategies. In this context, expressive volumes of data are used, requiring great effort to understand their structures, and then access information with the desired quality. This study are designed to provide an initial response to the need for mapping zones in the city of Itajaí (SC), Brazil. The solution proposes to aid object recognition employing object-based classifiers OneR, NaiveBayes, J48, IBk, and Hoeffding Tree algorithms used together with GeoDMA, and a first approach in the use of Region-based Convolutional Neural Network (R-CNN) and the YOLO algorithm. All this is to characterize vegetation zones, exposed soil zones, asphalt, and buildings within an urban and rural area. Through the implemented model for active identification of geospatial objects with similarity levels, it was possible to apply the data crossover after detecting the best classifier with accuracy (85%) and the kappa agreement coefficient (76%). The case study presents the dynamics of urban and rural expansion, where expressive volumes of data are obtained and submitted to different methods of cataloging and preparation to subsidize rapid control actions. Finally, the research describes a practical and systematic approach, evaluating the extraction of information to the recommendation of knowledge with greater scientific relevance. Allowing the methods presented to apply the calibration of values for each object, to achieve results with greater accuracy, which is proposed to help improve conservation and management decisions related to the zones within the city, leaving as a legacy the construction of a minimum technological infrastructure to support the decision.

Cells Responding to Closely Related Cholesterol-Dependent Cytolysins Release Extracellular Vesicles with a Common Proteomic Content Including Membrane Repair Proteins.

The plasma membrane (PM) protects cells from extracellular threats and supports cellular homeostasis. Some pathogens produce pore-forming toxins (PFTs) that disrupt PM integrity by forming transmembrane pores. High PFT concentrations cause massive damage leading to cell death and facilitating infection. Sub-lytic PFT doses activate repair mechanisms to restore PM integrity, support cell survival and limit disease. Shedding of extracellular vesicles (EVs) has been proposed as a key mechanism to eliminate PFT pores and restore PM integrity. We show here that cholesterol-dependent cytolysins (CDCs), a specific family of PFTs, are at least partially eliminated through EVs release, and we hypothesize that proteins important for PM repair might be included in EVs shed by cells during repair. To identify new PM repair proteins, we collected EVs released by cells challenged with sub-lytic doses of two different bacterial CDCs, listeriolysin O and pneumolysin, and determined the EV proteomic repertoire by LC-MS/MS. Intoxicated cells release similar EVs irrespectively of the CDC used. Also, they release more and larger EVs than non-intoxicated cells. A cluster of 70 proteins including calcium-binding proteins, molecular chaperones, cytoskeletal, scaffold and membrane trafficking proteins, was detected enriched in EVs collected from intoxicated cells. While some of these proteins have well-characterized roles in repair, the involvement of others requires further study. As proof of concept, we show here that Copine-1 and Copine-3, proteins abundantly detected in EVs released by intoxicated cells, are required for efficient repair of CDC-induced PM damage. Additionally, we reveal here new proteins potentially involved in PM repair and give new insights into common mechanisms and machinery engaged by cells in response to PM damage.

Comparative effect of dense-and-disperse versus non-repetitive and non-sequential frequencies in electroacupuncture-induced analgesia in a rodent model of peripheral neuropathic pain.

BACKGROUND: Neuropathic pain (NP) is a complex disease that remains challenging to treat. Low-frequency dense-and-disperse (DD) electroacupuncture (EA) has been used as adjuvant therapy for neuropathic pain; however, its analgesic effect decreases as stimulation time increases, or when it is repeatedly used. We hypothesized that a new frequency parameter could improve the effectiveness of EA, and aimed to compare the efficacy and duration of the analgesic effect between classic DD-EA and non-repetitive and non-sequential frequency (random frequency (RF)-EA) in neuropathic rats. Furthermore, the effect of RF-EA at local traditional acupuncture point locations versus auricular vagus nerve stimulation (aVNS) was evaluated. METHODS: Male Wistar rats with peripheral neuropathy were subjected to a single session of DD-EA or RF-EA for 20 or 40 min at ST36 + GB34. An additional group of rats was treated with RF-EA for 20 min using aVNS at the appropriate ear point locations. Paw pressure test, von Frey filaments and spontaneous pain scores were evaluated. Sham-operated rats were used as controls. RESULTS: In all, 20 min of RF-EA reversed hyperalgesia (for 24 h) and allodynia (for 8 h), showing a longer analgesic effect than DD-EA. Both RF-EA and DD-EA induced partial inhibition of spontaneous pain for 8 h. Forty minutes of DD-EA did not interfere with the NP phenomena; however, RF-EA induced significant long-term analgesia. aVNS induced an analgesic effect similar to local stimulation. CONCLUSION: This pilot study shows that RF-EA at both local traditional acupuncture point and auriculotherapy point locations induces long-lasting analgesia in neuropathic rats, and more effectively so than classical DD-EA.