Dynamics of ecosystem services and nonlinear responses to increased anthropogenic pressure.

Escalating global human activities elicit diverse ecosystem service responses, yet understanding remains limited. This study establishes a framework to clarify these responses, focusing on the Yangtze River Economic Belt in China. Analyzing 2000-2020 data, it calculates ecosystem service economic value and human footprint index. It introduces the ecosystem services response index and comprehensive responsiveness index to assess response characteristics and intensity to anthropogenic pressures. Results show a fluctuating decline in ecosystem services and an increase in anthropogenic pressures. There is a nonlinear relationship: ecosystem services decline with rising pressures, following a U-shaped trend. Notably, nonurban agglomerations experience more significant ecosystem service evolution than urban agglomerations due to differing environmental conditions. This highlights regional disparities in human activity impacts on ecosystems, crucial for planning.

Single-agent metronomic versus weekly oral vinorelbine as first-line chemotherapy in patients with HR-positive/HER2-negative advanced breast cancer: The randomized Tempo Breast study.

INTRODUCTION: Single-agent oral vinorelbine is a standard of care for hormone receptor (HR)-positive/human epidermal growth factor receptor 2 (HER2)-negative advanced breast cancer (ABC) that has progressed on endocrine therapy. Metronomic administration may offer a better balance of efficacy and safety than standard regimens, but data from previous trials are scarce. METHODS: In this open-label, multicenter, phase II trial, patients were randomized to oral vinorelbine administered on a metronomic (50 mg three times weekly) or weekly (60 mg/m2 in cycle 1, increasing to 80 mg/m2 if well tolerated) schedule. Treatment was continued until disease progression or intolerance. The primary endpoint was disease control rate (DCR, the proportion of patients with a best overall confirmed response of CR, PR, or stable disease lasting 6 months or more). RESULTS: One-hundred sixty-three patients were randomized and treated. The DCR was 63.4% (95% confidence interval [CI]: 52.0-73.8) with metronomic vinorelbine and 72.8% (95% CI: 61.8-82.1) with weekly vinorelbine. Weekly vinorelbine was also associated with longer progression-free survival (5.6 vs 4.0 months) and overall survival (26.7 vs 22.3 months) than metronomic vinorelbine, but was associated with more adverse events. CONCLUSIONS: In this randomized phase II trial, single-agent metronomic oral vinorelbine was effective and well tolerated as first-line chemotherapy for patients with HR-positive/HER2-negative ABC. Formal comparisons are not done in this phase II study and one can simply observe that confidence intervals of all endpoints overlap. When deciding for a chemotherapy after failure of endocrine therapy and CDK 4/6 inhibitors, oral vinorelbine might be an option to be given with either schedule. CLINICAL TRIAL REGISTRATION NUMBER: EudraCT 2014-003860-19.

Country-specific net-zero strategies of the pulp and paper industry.

The pulp and paper industry is an important contributor to global greenhouse gas emissions1,2. Country-specific strategies are essential for the industry to achieve net-zero emissions by 2050, given its vast heterogeneities across countries3,4. Here we develop a comprehensive bottom-up assessment of net greenhouse gas emissions of the domestic paper-related sectors for 30 major countries from 1961 to 2019-about 3.2% of global anthropogenic greenhouse gas emissions from the same period5-and explore mitigation strategies through 2,160 scenarios covering key factors. Our results show substantial differences across countries in terms of historical emissions evolution trends and structure. All countries can achieve net-zero emissions for their pulp and paper industry by 2050, with a single measure for most developed countries and several measures for most developing countries. Except for energy-efficiency improvement and energy-system decarbonization, tropical developing countries with abundant forest resources should give priority to sustainable forest management, whereas other developing countries should pay more attention to enhancing methane capture rate and reducing recycling. These insights are crucial for developing net-zero strategies tailored to each country and achieving net-zero emissions by 2050 for the pulp and paper industry.

Demographic Diversity of Clinical Trials for Therapeutic Drug Products: A Systematic Review of Recently Published Articles, 2017-2022.

The US Food and Drug Administration (FDA) encourages the use of enrollment practices that will lead to clinical trials that reflect the population most likely to use the therapeutic product (drug or biologic), if approved. In doing so, the benefit-risk profile of the product may be assessed more completely and offer patients and their health care providers a better understanding of the drug profile and greater confidence in clinical trial results. The objective of this systematic review was to assess recent literature on the demographic diversity of clinical trial participants, describe the methods used in defining clinical trial diversity, and address knowledge gaps to enhance clinical trial diversity. Our literature search initially yielded 246 articles. After applying our eligibility criteria, we conducted a full-text review and analyzed the contents of the 28 remaining articles in our systematic review. Eleven (39%) of the 28 articles used a reference standard to compare the participation of populations in clinical trials to assess diversity. The majority of the 28 articles reported on adult participants; only 5 included pediatric populations. Most articles found that women and minority populations were underrepresented in clinical trials. Some articles proposed solutions to improve clinical trial diversity; however, several did not comment on clinical trial diversity. Despite a growing emphasis on demographic diversity in research, certain populations continue to be underrepresented in clinical trials. There is a need to standardize the definition of diversity in clinical trials. Future research into effective enrollment approaches and appropriate reference standards could improve demographic diversity.

Lipid compartments and lipid metabolism as therapeutic targets against coronavirus.

Lipids perform a series of cellular functions, establishing cell and organelles' boundaries, organizing signaling platforms, and creating compartments where specific reactions occur. Moreover, lipids store energy and act as secondary messengers whose distribution is tightly regulated. Disruption of lipid metabolism is associated with many diseases, including those caused by viruses. In this scenario, lipids can favor virus replication and are not solely used as pathogens' energy source. In contrast, cells can counteract viruses using lipids as weapons. In this review, we discuss the available data on how coronaviruses profit from cellular lipid compartments and why targeting lipid metabolism may be a powerful strategy to fight these cellular parasites. We also provide a formidable collection of data on the pharmacological approaches targeting lipid metabolism to impair and treat coronavirus infection.

High-frequency sectoral carbon and environmental analysis based on monthly input-output tables compilation during 2018-2021.

This paper proposes a process for updating monthly input-output tables with monthly macroeconomic statistics and published input-output tables. Reasonable assumptions are set up and 48 monthly input-output tables are prepared from 2018 to 2021 with the combination of the row range series method and nonlinear mathematical planning. The Weaver-Thomas composite index is used to analyze the role of the sector in the economic network, and the sectoral correlation indicators are used to analyze the correlation change of the sector's monthly electricity emissions to show an environmental application effect of the monthly input-output table. The results show that the monthly input-output tables can be prepared with acceptable accuracy, and they can reveal the sectoral network structure changes and sectoral carbon emissions changes in continuous monthly time series. The proposed approach contributes for the compilation of high-time-frequency input-output tables, so as to support high-frequency industrial environmental impact analysis.

Lipid droplets in Zika neuroinfection: Potential targets for intervention?

Lipid droplets (LD) are evolutionarily conserved lipid-enriched organelles with a diverse array of cell- and stimulus-regulated proteins. Accumulating evidence demonstrates that intracellular pathogens exploit LD as energy sources, replication sites, and part of the mechanisms of immune evasion. Nevertheless, LD can also favor the host as part of the immune and inflammatory response to pathogens. The functions of LD in the central nervous system have gained great interest due to their presence in various cell types in the brain and for their suggested involvement in neurodevelopment and neurodegenerative diseases. Only recently have the roles of LD in neuroinfections begun to be explored. Recent findings reveal that lipid remodelling and increased LD biogenesis play important roles for Zika virus (ZIKV) replication and pathogenesis in neural cells. Moreover, blocking LD formation by targeting DGAT-1 in vivo inhibited virus replication and inflammation in the brain. Therefore, targeting lipid metabolism and LD biogenesis may represent potential strategies for anti-ZIKV treatment development. Here, we review the progress in understanding LD functions in the central nervous system in the context of the host response to Zika infection.

How robust are current narratives to deal with the urban energy-water-land nexus?

Current energy, water, and land (EWL) nexus research treats all resources equally, causing bias in complicated nexus studies. To make the analysis robust, we consider resource endowment and significance. Here, we provide a methodological framework where the urban industrial resource nexus strength is constructed and assign weights to resources according to policies, describing resource efficiency and representing it in ternary diagrams to assess the urban industrial nexus innovatively. Results showed that energy drives urban development under all weights, with energy resource efficiency exceeding 60%. From consumption-based accounting, energy continues to dominate most industries under physical weightings but emphasizes the significance of water and land. While, under economic weightings, land supplants energy's dominance in specific sectors. Setting weights helps understand resource interaction, establish synergy based on urban development objectives, and minimize robustness. Our findings provide quantitative evidence for assessing urban resource efficiency to highlight priority sectors for intervention in urban decision-making.

Patients´ satisfaction concerning direct anterior dental restoration.

The objective of this study was to observe patients' satisfaction with their in-service direct anterior dental restorations and to compare it with clinical evaluation using FDI (Federation Dental International) criteria. Patients scored their own anterior dental restorations regarding satisfaction (satisfactory /dissatisfactory). If dissatisfaction was mentioned, then, they would be interviewed about the complaint. In the same session, the dental restorations were clinically evaluated by two dentists using FDI criteria (1-5 score) concerning esthetic, functional, and biological domains. Descriptive statistics were used for frequencies of scores attributed by patients and clinicians. In order to compare patients' to clinicians' frequencies, the Chi-square test was applied (p ≤ 0.05). A total of 106 restorations were evaluated by patients and clinicians. Patients reported 52.8% of restorations satisfactory and 47.8% dissatisfactory. Overall, clinicians reported the same restorations as 82,3% satisfactory and 17,6% dissatisfactory. Patients' most frequent complaints referred to color, followed by anatomical form, fracture of material and retention, and approximal anatomical form. Comparing patients' satisfaction and dissatisfaction rates to clinicians' evaluation per criteria, there was no difference regarding esthetics. The frequency of dissatisfactory restorations by clinicians was significantly lower when functional and biological properties were compared with patients' opinions. Direct anterior dental restorations were more frequently reported as satisfactory by patients and clinicians, being the main complaints related to esthetic issues. When clinicians and patients' evaluations were compared, it was observed that the frequencies of satisfactory restoration by patients and clinicians were similar regarding esthetic properties, and significantly different regarding functional and biological properties.

A multi-criteria approach to assess interconnections among the environmental, economic, and social dimensions of circular economy.

The debate about the negative impacts that production and consumption cause on the environment is in vogue. Strategies that point to a sustainable, healthy, and resilient path are being sought. One of these paths is the Circular Economy, which emerges as an alternative to reduce the socio-environmental impacts caused by the linear model of production-use-disposal, presenting opportunities to generate revenue, income, and wealth with circular processes. However, despite the circular economy being considered an essential strategy to improve overall performance toward sustainability in its three dimensions, recent research has shown that the predominant focus of circular approaches is on the economic and environmental dimensions. At the same time, the social aspects still need to be explored. This article addresses this problem, aiming to explore circular economy environmental, economic, and social elements in three economic blocs from 2000 to 2020 using the Five Sector Sustainability Model, establishing a baseline to co-create an equitable and regenerative future. The results showed that ASEAN in 2000 occupied the first position in the general ranking. The European Union had the best classification in the economic sector, and Mercosur was the best regarding social benefits. In 2020, while the European bloc was better positioned in the general ranking, the South American bloc occupied the last post in almost all sectors. Comparatively, the highest-ranked bloc in 2020 in overall sustainability is more in line with the UN SDG due to circular actions oriented towards the three fundamental pillars of sustainability.

Patients´ satisfaction concerning direct anterior dental restoration

Abstract The objective of this study was to observe patients' satisfaction with their in-service direct anterior dental restorations and to compare it with clinical evaluation using FDI (Federation Dental International) criteria. Patients scored their own anterior dental restorations regarding satisfaction (satisfactory /dissatisfactory). If dissatisfaction was mentioned, then, they would be interviewed about the complaint. In the same session, the dental restorations were clinically evaluated by two dentists using FDI criteria (1-5 score) concerning esthetic, functional, and biological domains. Descriptive statistics were used for frequencies of scores attributed by patients and clinicians. In order to compare patients' to clinicians' frequencies, the Chi-square test was applied (p ≤ 0.05). A total of 106 restorations were evaluated by patients and clinicians. Patients reported 52.8% of restorations satisfactory and 47.8% dissatisfactory. Overall, clinicians reported the same restorations as 82,3% satisfactory and 17,6% dissatisfactory. Patients' most frequent complaints referred to color, followed by anatomical form, fracture of material and retention, and approximal anatomical form. Comparing patients' satisfaction and dissatisfaction rates to clinicians' evaluation per criteria, there was no difference regarding esthetics. The frequency of dissatisfactory restorations by clinicians was significantly lower when functional and biological properties were compared with patients' opinions. Direct anterior dental restorations were more frequently reported as satisfactory by patients and clinicians, being the main complaints related to esthetic issues. When clinicians and patients' evaluations were compared, it was observed that the frequencies of satisfactory restoration by patients and clinicians were similar regarding esthetic properties, and significantly different regarding functional and biological properties.

Resumo O objetivo deste estudo foi observar a satisfação dos pacientes com suas restaurações dentárias anteriores diretas e compará-las com a avaliação clínica do dentista usando os critérios FDI (Federation Dental International). Os pacientes pontuaram suas restaurações dentárias (n=106) anteriores em relação à satisfação (satisfatória / insatisfatória). Quando insatisfatória, ele foi entrevistado sobre a queixa. Na mesma sessão, as restaurações dentárias foram avaliadas clinicamente por dois dentistas utilizando os critérios FDI (escore 1-5) quanto aos aspectos estéticos, funcionais e biológicos. Estatística descritiva foi usada para frequências de escores atribuídos por pacientes e clínicos. Para comparar as frequências dos pacientes e dos clínicos, foi aplicado o teste Qui-quadrado (p ≤ 0,05). Os pacientes relataram suas restaurações como 52,8% satisfatórias e 47,8% insatisfatórias. Os clínicos reportaram as mesmas restaurações, 82,3% satisfatória e 17,6% insatisfatória. As queixas mais frequentes dos pacientes referiam-se à cor, seguida da forma anatômica, fratura e retenção do material e forma anatômica proximal. Comparando os índices de satisfação e insatisfação dos pacientes com os clínicos, não houve diferença em relação à estética. A frequência de restaurações insatisfatórias por dentistas foi significativamente menor quando as propriedades funcionais e biológicas foram comparadas com as opiniões dos pacientes. As restaurações foram mais frequentemente relatadas como satisfatórias pelos pacientes, sendo as principais queixas relacionadas a questões estéticas. Quando as avaliações dos clínicos e dos pacientes foram comparadas, observou-se que as frequências de restaurações satisfatórias por pacientes e clínicos foram semelhantes em relação às propriedades estéticas e significativamente diferentes em relação às propriedades funcionais e biológicas.

Burnout in Nurses of an Intensive Care Unit during COVID-19: A Pilot Study in Portugal.

BACKGROUND: This study aimed to evaluate burnout in intensive care unit nurses and describe the relationship between burnout dimensions and sociodemographic and professional variables. METHODS: A pilot study was carried out with 29 intensive care nurses during the COVID-19 pandemic. The data were collected using an online questionnaire consisting of a sociodemographic characterization and the Maslach Burnout Inventory Human Services Survey. In the comparative statistical analysis, the nonparametric Mann-Whitney and Kruskal-Wallis tests were used (p < 0.05). RESULTS: The Emotional Exhaustion dimension represented an average of 27.9 ± 10.4, and the Personal Fulfillment of 30.8 ± 7.0 was considered high. Regarding Depersonalization, medium-level results (8.1 ± 6.0) were found. There was a prevalence of 41.3% of high levels of burnout. There was no statistically significant difference in the comparison between the three dimensions and sociodemographic and professional variables (p > 0.05). CONCLUSIONS: The global results point to high levels of burnout in nurses, with greater expression in the Emotional Exhaustion and Professional Fulfillment domains. There were no differences between groups, demonstrating homogeneity in the team's level of burnout. The incidence of burnout in this study was higher than that identified in other studies carried out in Portugal with health professionals. This pilot study will allow the defining of strategies for reducing burnout and, consequently, a proposed extension of the study to other organization wards.

Metabolic reprogramming and lipid droplets are involved in Zika virus replication in neural cells.

Zika virus (ZIKV) infection is a global public health concern linked to adult neurological disorders and congenital diseases in newborns. Host lipid metabolism, including lipid droplet (LD) biogenesis, has been associated with viral replication and pathogenesis of different viruses. However, the mechanisms of LD formation and their roles in ZIKV infection in neural cells are still unclear. Here, we demonstrate that ZIKV regulates the expression of pathways associated with lipid metabolism, including the upregulation and activation of lipogenesis-associated transcription factors and decreased expression of lipolysis-associated proteins, leading to significant LD accumulation in human neuroblastoma SH-SY5Y cells and in neural stem cells (NSCs). Pharmacological inhibition of DGAT-1 decreased LD accumulation and ZIKV replication in vitro in human cells and in an in vivo mouse model of infection. In accordance with the role of LDs in the regulation of inflammation and innate immunity, we show that blocking LD formation has major roles in inflammatory cytokine production in the brain. Moreover, we observed that inhibition of DGAT-1 inhibited the weight loss and mortality induced by ZIKV infection in vivo. Our results reveal that LD biogenesis triggered by ZIKV infection is a crucial step for ZIKV replication and pathogenesis in neural cells. Therefore, targeting lipid metabolism and LD biogenesis may represent potential strategies for anti-ZIKV treatment development.

Lipid droplets in Zika neuroinfection: Potential targets for intervention?

Lipid droplets (LD) are evolutionarily conserved lipid-enriched organelles with a diverse array of cell- and stimulus-regulated proteins. Accumulating evidence demonstrates that intracellular pathogens exploit LD as energy sources, replication sites, and part of the mechanisms of immune evasion. Nevertheless, LD can also favor the host as part of the immune and inflammatory response to pathogens. The functions of LD in the central nervous system have gained great interest due to their presence in various cell types in the brain and for their suggested involvement in neurodevelopment and neurodegenerative diseases. Only recently have the roles of LD in neuroinfections begun to be explored. Recent findings reveal that lipid remodelling and increased LD biogenesis play important roles for Zika virus (ZIKV) replication and pathogenesis in neural cells. Moreover, blocking LD formation by targeting DGAT-1 in vivo inhibited virus replication and inflammation in the brain. Therefore, targeting lipid metabolism and LD biogenesis may represent potential strategies for anti-ZIKV treatment development. Here, we review the progress in understanding LD functions in the central nervous system in the context of the host response to Zika infection.

The Endolysosomal System: The Acid Test for SARS-CoV-2.

This review aims to describe and discuss the different functions of the endolysosomal system, from homeostasis to its vital role during viral infections. We will initially describe endolysosomal system's main functions, presenting recent data on how its compartments are essential for host defense to explore later how SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2) and other coronaviruses subvert these organelles for their benefit. It is clear that to succeed, pathogens' evolution favored the establishment of ways to avoid, escape, or manipulate lysosomal function. The unavoidable coexistence with such an unfriendly milieu imposed on viruses the establishment of a vast array of strategies to make the most out of the invaded cell's machinery to produce new viruses and maneuvers to escape the host's defense system.

Simvastatin Downregulates the SARS-CoV-2-Induced Inflammatory Response and Impairs Viral Infection Through Disruption of Lipid Rafts.

Coronavirus disease 2019 (COVID-19) is currently a worldwide emergency caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). In observational clinical studies, statins have been identified as beneficial to hospitalized patients with COVID-19. However, experimental evidence of underlying statins protection against SARS-CoV-2 remains elusive. Here we reported for the first-time experimental evidence of the protective effects of simvastatin treatment both in vitro and in vivo. We found that treatment with simvastatin significantly reduced the viral replication and lung damage in vivo, delaying SARS-CoV-2-associated physiopathology and mortality in the K18-hACE2-transgenic mice model. Moreover, simvastatin also downregulated the inflammation triggered by SARS-CoV-2 infection in pulmonary tissue and in human neutrophils, peripheral blood monocytes, and lung epithelial Calu-3 cells in vitro, showing its potential to modulate the inflammatory response both at the site of infection and systemically. Additionally, we also observed that simvastatin affected the course of SARS-CoV-2 infection through displacing ACE2 on cell membrane lipid rafts. In conclusion, our results show that simvastatin exhibits early protective effects on SARS-CoV-2 infection by inhibiting virus cell entry and inflammatory cytokine production, through mechanisms at least in part dependent on lipid rafts disruption.

The effects of early rehabilitation on functional exercise tolerance in decompensated heart failure patients: Results of a multicenter randomized controlled trial (ERIC-HF study).

OBJECTIVE: To analyze (1) the effect of an aerobic training program on functional exercise tolerance in decompensated heart failure (DHF) patients; (2) to assess the effects of an aerobic training program on functional independence; and (3) dyspnea during activities of daily living. DESIGN: A randomized controlled clinical trial with follow-up at discharge. SETTINGS: Eight hospitals. Recruitment took place between 9/ 2017 and 3/2019. GROUP ASSIGNMENTS: Patients with DHF who were admitted to the hospital, were randomly assigned to usual rehabilitation care guideline recommended (control group) or aerobic training program (exercise group). MAIN OUTCOME: Functional exercise tolerance was measured with a 6-min walking test at discharge. RESULTS: In total 257 patients with DHF were included, with a mean age of 67 ± 11 years, 84% (n = 205) had a reduced ejection fraction and the hospital stay was 16 ± 10 days. At discharge, patients in the intervention group walked further compared to the control group (278 ± 117m vs 219 ± 115m, p < 0.01) and this difference stayed significant after correcting for confounders (p < 0.01). A significant difference was found favoring the exercise group in functional independence (96 ± 7 vs 93 ± 12, p = 0.02) and dyspnea associated to ADL (13 ± 5 vs 17 ± 7, p < 0.01) and these differences persisted after correcting for baseline values and confounders (functional independence p < 0.01; dyspnea associated with ADL p = 0.02). CONCLUSION: The ERIC-HF program is safe, feasible, and effective in increasing functional exercise tolerance and functional independence in hospitalized patients admitted due to DHF.

Energy constrains to increasing complexity in the biosphere.

Thirty years ago, the systems ecologist Howard T. Odum introduced the concept of transformity, which is a thermodynamic measure of quality within the trial and error evolutionary dynamics of ecosystems, namely an indicator of rank in the hierarchical system structure of the biosphere. Based on a global database of individual processes and whole economies, this paper extends, refines, and updates Odum's idea, demonstrating the strength of the postulated relation. In particular, an inverse linear logarithmic relationship is shown to hold between resource quantity (exergy) and quality (emergy), which is the result of an overall energetic efficiency characteristic of energy transformation processes of the biosphere. This relation extends from natural renewable energy sources to human information (including global internet data flows) and know-how embedded in national economies, thus identifying a consistent theory of hierarchical organization of the biosphere grounded in energetics and ultimately setting constraints to illusions of unlimited growth.

Clinical decision-making in anterior resin composite restorations: a multicenter evaluation.

OBJECTIVE: This study investigated whether a sample of anterior resin composite restorations could be differently evaluated in different centers of evaluation by clinical and lay evaluators. METHODS: Anterior resin composite restorations on high-quality intraoral digital photography were evaluated using FDI criteria (1-5 score) by pairs of clinical and lay evaluators in Brazil (BR), France (FR), Peru (PE), and the United States of America (USA). Scores were allocated as maintenance (1, 2, 3), repair (4) and replacement (5) when comparing clinical evaluators and, as acceptable (1, 2, 3) and unacceptable (4, 5) when comparing clinical vs. lay evaluators and lay vs. lay evaluators. The Chi-square test compared the frequencies of scores among the centers. RESULTS: The frequencies of maintaining, repairing, or replacing anterior resin composite restorations given by clinical evaluators varied depending on the evaluation center. BR and PE showed the highest frequencies for repair and replacement, while FR and USA showed the highest frequencies for maintenance. The comparison of frequencies of anterior resin composite restorations accepted or unaccepted by the clinical vs lay evaluators in the same centers showed a significantly higher frequency of acceptable dental restorations coming from clinical evaluators. Comparison between lay evaluators from different centers showed significant higher frequency of unacceptable dental restorations by BR, compared to other centers. CONCLUSIONS: In the evaluation of anterior resin composite restorations, the maintenance, repair, or replacement trends can vary among different centers. The unacceptable rate came more frequently from lay than from clinical evaluators. Lay evaluators from different centers differed significantly. CLINICAL SIGNIFICANCE: Clinical and lay evaluators in distant evaluation centers can present different trends when assessing anterior resin composite restoration. Multicenter evaluations can help to understand such differences and it is important because clinical decision-making based on scientific evidence comes from clinical studies done in different research centers.