bluuebunny/crossref_metadata_2025 huggingface dataset:
Design of Feedback Control Strategies in a Plant-Wide Wastewater Treatment Plant for Simultaneous Evaluation of Economics, Energy Usage, and Removal of Nutrients
Authors: Sheik Abdul , Tejaswini Eagalapati , Seepana Murali , Ambati Seshagiri , Meneses Montse , Vilanova Ramon
Published: October 2021

Simultaneous removal of nitrogen and phosphorous is a recommended practice while treating wastewater. In the present study, control strategies based on proportional-integral (PI), model predictive control (MPC), and fuzzy logic are developed and implemented on a plant-wide wastewater treatment plant. Four combinations of control frameworks are developed in order to reduce the operational cost and improve the effluent quality. As a working platform, a Benchmark simulation model (BSM2-P) is used. A default control framework with PI controllers is used to control nitrate and dissolved oxygen (DO) by manipulating the internal recycle and oxygen mass transfer coefficient (KLa). Hierarchical control topology is proposed in which a lower-level control framework with PI controllers is implemented to DO in the sixth reactor by regulating the KLa of the fifth, sixth, and seventh reactors, and fuzzy and MPC are used at the supervisory level. This supervisory level considers the ammonia in the last aerobic reactor as a feedback signal to alter the DO set-points. PI-fuzzy showed improved effluent quality by 21.1%, total phosphorus removal rate by 33.3% with an increase of operational cost, and a slight increase in the production rates of greenhouse gases. In all the control design frameworks, a trade-off is observed between operational cost and effluent quality.
Commercial Energy Demand Forecasting in Bangladesh
Authors: Anik Asif Reza , Rahman Sanzidur
Published: October 2021

Although both aggregate and per capita energy consumption in Bangladesh is increasing rapidly, its per capita consumption is still one of the lowest in the world. Bangladesh gradually shifted from petroleum-based energy to domestically sourced natural-gas-based energy sources, which are predicted to run out within next two decades. The present study first identified the determinants of aggregate commercial energy and its three major components of oil, natural gas, and coal demand for Bangladesh using a simultaneous equations framework on an annual database covering a period of 47 years (1972–2018). Next, the study forecast future demand for aggregate commercial energy and its three major components for the period of 2019–2038 under the business-as-usual and ongoing COVID-19 pandemic scenarios with some assumptions. As part of a sensitivity analysis, based on past trends, we also hypothesized four alternative GDP and population growth scenarios and forecast corresponding changes in total energy demand forecast. The results revealed that while GDP and lagged energy demand are the major drivers of energy demand in the country, we did not see strong effects of own- and cross-price elasticities of energy sources, which we attributed to three reasons: subsidized low energy prices, time and cost required to switch between different energy-mix technologies, and suppressed energy demand. The aggregate energy demand is expected to increase by 400% by the end of the forecasting period in 2038 from its existing level in 2018 under the business-as-usual scenario, whereas the effect of COVID-19 could suppress it down to 300%. Under the business-as-usual scenario, the highest increase will occur for coal (3.94-fold), followed by gas (2.64-fold) and oil (2.37-fold). The COVID-19 pandemic will suppress the future demand of all energy sources at variable rates. The ex ante forecasting errors were small, varying within the range of 3.6–3.7% of forecast values. Sensitivity analysis of changes in GDP and population growth rates showed that forecast total energy demand will increase gradually from 3.58% in 2019 to 8.79% by 2038 from original forecast values. Policy recommendations include capacity building of commercial energy sources while ensuring the safety and sustainability of newly proposed coal and nuclear power installations, removing inefficiency of production and distribution of energy and its services, shifting towards renewable and green energy sources (e.g., solar power), and redesigning subsidy policies with market-based approaches.
The Importance of Extraction Protocol on the Analysis of Novel Waste Sources of Lignocellulosic Biomass
Authors: Hickey Daniel T. , Hayes Daniel J. , Pembroke J. Tony , Ryan Michael P. , Leahy James J.
Published: October 2021

As the utilization and consumption of lignocellulosic biomass increases, so too will the need for an adequate supply of feedstock. To meet these needs, novel waste feedstock materials will need to be utilized. Exploitation of these novel feedstocks will require information both on the effects of solvent extraction on the succeeding analysis of potential novel feedstocks and how accurate current methodologies are in determining the composition of novel lignocellulosic feedstocks, particularly the carbohydrate and lignin fractions. In this study, the effects of solvent extraction on novel feedstocks, including tree foliage, tree bark and spent mushroom compost, with 95% ethanol, water and both sequentially were examined. Chemical analyses were carried out to determine the moisture content, ash, extractives, post-hydrolysis sugars, Klason lignin (KL) and acid-soluble lignin (ASL) within the selected feedstocks. The result of extraction could be seen most strongly for Klason lignin, with a strong association between higher levels of Klason lignin levels and greater amounts of non-removed extractives (tree foliage and bark). Higher Klason lignin levels are reported to be due the condensation of non-removed extractives during hydrolysis, hence the lower Klason lignin determinations following extraction are more exact. In addition, total sugar determinations were lower following extractions. This is because of the solubility of non-cell-wall carbohydrates; thus, the determinations following extraction are more accurate representations of structural cell-wall polysaccharides such as cellulose. Such determinations will assist in determining the best way to utilize novel feedstocks such as those analyzed in this work.
Maternal Psychological Distress and Children’s Internalizing/Externalizing Problems during the COVID-19 Pandemic: The Moderating Role Played by Hypermentalization
Authors: Bianco Federica , Levante Annalisa , Petrocchi Serena , Lecciso Flavia , Castelli Ilaria
Published: October 2021

In order to explore the psychological impact of the COVID-19 outbreak on the caregiver–child relationship, we investigated the interplay among COVID-19 exposure and children’s internalizing/externalizing problems during the Italian lockdown, hypothesizing a mediation effect played by maternal distress. Additionally, we included maternal reflective functioning (i.e., hypermentalization) as a moderator factor among this interplay. A total of 305 Italian mothers of children aged 6–13 years (M = 10.3; SD = 2.4) filled in an online survey. Findings revealed an indirect effect of maternal COVID-19 exposure on children’s anxious/depressed (k2 = 0.46) and attention problems (k2 = 0.32) via maternal distress. Hypermentalization moderated the impact of maternal COVID-19 exposure on children’s anxious/depressed problems (β = −1.08, p = 0.04). Hypermentalization moderated both the relation between maternal distress and children’s aggressive behaviors (β = 12.226; p < 0.001) and between maternal distress and children’s attention problems (β = 5.617, p < 0.001). We found pivotal significant effects of maternal hypermentalization on children’s anxious/depressed and attention problems, indicating that the higher the mother’s hypermentalization was, the higher the children’s problems were. Our results broaden what we knew on the role of maternal reflective and emotional functioning on children’s emotional/behavioral adjustment during stressful situations.
Ergonomic Comparison of Four Dental Workplace Concepts Using Inertial Motion Capture for Dentists and Dental Assistants
Authors: Ohlendorf Daniela , Fraeulin Laura , Haenel Jasmin , Betz Werner , Erbe Christina , Holzgreve Fabian , Wanke Eileen M. , Brueggmann Doerthe , Nienhaus Albert , Maurer-Grubinger Christian , Groneberg David A.
Published: October 2021

When the inventory is arranged in a dental practice, a distinction can be made between four different dental workplace concepts (DWCs). Since the prevalence of musculoskeletal diseases in dental professionals is very high, preventive solution need to be investigated. As the conventionally used DWCs have, to date, never been studied in terms of their ergonomics, this study aims to investigate the ergonomic risk when working at the four different DWCs. In total, 75 dentists (37 m/38 f) and 75 dental assistants (16 m/59 f) volunteered to take part in this study. Standardized cooperative working procedures were carried out in a laboratory setting and kinematic data were recorded using an inertial motion capture system. The data were applied to an automated version of the Rapid Upper Limb Assessment (RULA). Comparisons between the DWCs and between the dentists and dental assistants were calculated. In all four DWCs, both dentists and dental assistants spent 95–97% of their working time in the worst possible RULA score. In the trunk, DWCs 1 and 2 were slightly favorable for both dentists and dental assistants, while for the neck, DWC 4 showed a lower risk score for dentists. The ergonomic risk was extremely high in all four DWCs, while only slight advantages for distinct body parts were found. The working posture seemed to be determined by the task itself rather than by the different inventory arrangements.
Facing the National Recovery and Resilience Plan: Sources of Data, Indicators, and Participatory Strategies in Healthcare and Social Fields
Authors: Franchini Michela , Molinaro Sabrina , Caiolfa Michelangelo , Salvatori Massimiliano , Pieroni Stefania
Published: October 2021

Innovation in governance and services should be the target of the Italian National Recovery and Resilience Plan. Monitoring processes, impacts, and outcomes requires a system of new indicators that are practical to collect. Secondary data sources, their availability, and their information potential should be evaluated, and primary sources should be implemented to supplement traditional disease surveillance. This work highlights the most relevant aspects for bridging the mismatching between complex community needs and current health/social supply and how those aspects could be faced. As a result, we propose a structured multi-phases process for setting the design and functionalities of a cooperative information system, built on the integration between secondary and primary data for informing policies about chronic low back pain (CLBP), a widely recognized determinant of disability and significant economic burden. In particular, we propose the Dress-KINESIS, a tool for improving community capacity development and participation that allows one to freely collect big health and social data and link it to existing secondary data. The system also may be able to monitor how the resources are distributed across different care sectors and suggest how to improve efficiency based on the patient’s CLBP risk stratification. Moreover, it is potentially customizable in other fields of health.
An Exploratory Survey of Mental Health Social Work in Europe
Authors: Stone Kevin , McCusker Pearse , Davidson Gavin , Vicary Sarah
Published: October 2021

This article reports on an exploratory study comparing mental health social work (MHSW) in Europe. There has been very limited previous research comparing approaches to MHSW in Europe and so the aim of the study was to develop a better understanding of the similarities and differences between and, where relevant, within countries (referred to as jurisdictions). An online survey was distributed mainly through existing European networks and social media to seek information on the role, nature, extent and context of MHSW in a range of European jurisdictions. Conducted during the COVID-19 pandemic, there were 158 responses from 10 jurisdictions. Data were analyzed using thematic analysis. From this analysis, four main themes were identified, relating to: role; law, policy and education; the distinctive contribution made by MHSW; and the key challenges for MHSW. The study demonstrates that MHSW, although it is described and provided in different ways and is confined by a range of factors, plays an important role in mental health services across jurisdictions. There are also interesting differences between contexts, especially in the balance of therapeutic, legal and specialist/generic approaches, some of which reflect the World Health Organisation’s vision and objectives for mental health. Lastly, the study illustrates a need and provides a valuable basis for further comparative and collaborative work to define MHSW and enhance the contributions it makes.
Predicting Fitness Centre Dropout
Authors: Sobreiro Pedro , Guedes-Carvalho Pedro , Santos Abel , Pinheiro Paulo , Gonçalves Celina
Published: October 2021

The phenomenon of dropout is often found among customers of sports services. In this study we intend to evaluate the performance of machine learning algorithms in predicting dropout using available data about their historic use of facilities. The data relating to a sample of 5209 members was taken from a Portuguese fitness centre and included the variables registration data, payments and frequency, age, sex, non-attendance days, amount billed, average weekly visits, total number of visits, visits hired per week, number of registration renewals, number of members referrals, total monthly registrations, and total member enrolment time, which may be indicative of members’ commitment. Whilst the Gradient Boosting Classifier had the best performance in predicting dropout (sensitivity = 0.986), the Random Forest Classifier was the best at predicting non-dropout (specificity = 0.790); the overall performance of the Gradient Boosting Classifier was superior to the Random Forest Classifier (accuracy 0.955 against 0.920). The most relevant variables predicting dropout were “non-attendance days”, “total length of stay”, and “total amount billed”. The use of decision trees provides information that can be readily acted upon to identify member profiles of those at risk of dropout, giving also guidelines for measures and policies to reduce it.
Associations between Longitudinal Patterns of Substance Use and Anxiety and Depression Symptoms among a Sample of Canadian Secondary School Students
Authors: Williams Gillian C. , Patte Karen A. , Ferro Mark A. , Leatherdale Scott T.
Published: October 2021

The objective of this study is to examine the longitudinal associations between latent classes of substance use and anxiety and depression scores among youth who use substances. This study uses data from three waves (Wave 1: 2017/18, Wave 2: 2018/19, and Wave 3: 2019/20) of the COMPASS study. Students in grades 9 and 10 who reported substance use at baseline (n = 738) report their substance use (alcohol, cannabis, cigarettes, and e-cigarettes) and anxiety and depression symptoms at each wave. A Repeated Measures Latent Class Analysis (RMLCA) is used to determine substance use classes, and mixed models are used to examine the associations between substance use classes and anxiety and depression. We identify three classes of substance use: (1) occasional alcohol and e-cigarette use, (2) escalating poly-substance use, and (3) consistent poly-substance use. After controlling for relevant covariates, consistent poly-substance use is associated with depression (Female OR: 1.24 [95%CI: 0.46, 2.02]; Male OR 1.13 [95%CI: 0.38, 1.87]) but not anxiety. Escalating poly-substance use is associated with depression among males (OR 0.72 [95%CI: 0.10, 1.33]). These findings should be taken into consideration when creating prevention programming and treatment strategies for adolescents. Substance use programming should be comprehensive, consider multiple substances, and be cognizant of symptoms of mental illness, particularly depression.
Pathway of Green Development of Yangtze River Economics Belt from the Perspective of Green Technological Innovation and Environmental Regulation
Authors: Ye Tifang , Zheng Hao , Ge Xiangyu , Yang Keling
Published: October 2021

The eco-efficiency of the Yangtze River Economic Belt from 2005 to 2019 has been evaluated by the super-efficiency SBM window model, the results of which are taken as the measurement standard for green development. Next, more attempts have been done to figure out the impacts of green technological innovation on the green development in urban clusters of the Yangtze River Economic Belt by a systematic GMM model, further confirming the moderation effect of dual environmental regulations on the relationship between green technological innovation and green development and the heterogeneity in different urban clusters of the Yangtze River Economic Belt. Finally, it is a fact that a cascade has been pointed out in green development of the Yangtze River Economic Zone. With an empirical analysis, it has been found that green technological innovation has a positive moderating effect on green development in the downstream regions, and the relationship between green technological innovation and green development is positively affected by the formal environmental regulations in the overall Yangtze River Economic Zone and the midstream region. Meanwhile, informal environmental regulations play a positive role in moderating the links between green technological innovation and green development in the overall Yangtze River Economic Zone, as well as the upstream, midstream and downstream regions. Based on the conclusions of the research, some policy suggestions of a multi-environmental regulation governance system and regional differentiated environmental regulation are given at last.
Public Perceptions of the Role of Lifestyle Factors in Cancer Development: Results from the Spanish Onco-Barometer 2020
Authors: Petrova Dafina , Borrás Josep Maria , Pollán Marina , Bayo Lozano Eloísa , Vicente David , Jiménez Moleón José Juan , Sánchez Maria José
Published: October 2021

The European Code against Cancer recommends not to smoke, to avoid alcohol consumption, to eat a healthy diet, and maintain a healthy weight to prevent cancer. To what extent is the public aware of the influence of these lifestyle factors on cancer development? The goal of the current study was to describe the perceived influence of four lifestyle factors (tobacco, alcohol, diet, and weight) on cancer development in the general population and identify factors related to low perceptions of influence. We analyzed data from the 2020 Onco-barometer (n = 4769), a representative population-based survey conducted in Spain. With the exception of smoking, lifestyle factors were among those with the least perceived influence, more so among the demographic groups at higher risk from cancer including men and older individuals (65+ years). Individuals from lower socio-economic groups were more likely to report not knowing what influence lifestyle factors have on cancer. Lower perceived influence was also consistently related to perceiving very low risk from cancer. Overall, although there is variation in perceptions regarding the different lifestyle factors, low perceived influence clusters among those at higher risk for cancer. These results signal the need for public health campaigns and messages informing the public about the preventive potential of lifestyle factors beyond avoiding tobacco consumption.
Effect of Phenol Application Time in the Treatment of Onychocryptosis: A Randomized Double-Blind Clinical Trial
Authors: Muriel-Sánchez Juan Manuel , Coheña-Jiménez Manuel , Montaño-Jiménez Pedro
Published: October 2021

Background: In the treatment of Onychocryptosis, chemical matricectomy with 88% phenol solution is one of the most common surgical procedures due to a recurrence rate of less than 5%, but it may produce a delay in healing time. The objective was to compare the healing time between phenol applications of 30 or 60 s. Methods: A comparative, prospective, parallel, randomized, and blinded clinical trial was registered with the European Clinical Trials Database. Twenty-seven patients (54 feet) with 108 affected nail folds were randomized and treated with chemical matricectomy with phenol. Each hallux was randomly assigned to one of two groups (60 vs. 30 s phenolization). Each patient and one investigator were blinded to the phenol application time in each foot. The outcome measurements were healing time, recurrence, pain, post-surgical bleeding, inflammation, and infection rate. Results: The 30 s application presents a shorter healing time (14.93 ± 2.81 days vs. 22.07 ± 3.16 days; p < 0.001) with a similar recurrence rate (p = 0.99). Post-operatory bleeding, pain, inflammation, and the infection rate did not show significant differences (p > 0.05). Conclusions: The 30 s phenol application time offers a shorter healing time than 60 s without affecting the effectiveness of the procedure, showing the same rate of complications.
The Mediation Effects of Aluminum in Plasma and Dipeptidyl Peptidase Like Protein 6 (DPP6) Polymorphism on Renal Function via Genome-Wide Typing Association
Authors: Chen Ting-Hao , Yang Chen-Cheng , Luo Kuei-Hau , Dai Chia-Yen , Chuang Yao-Chung , Chuang Hung-Yi
Published: October 2021

Aluminum (Al) toxicity is related to renal failure and the failure of other systems. Although there were some genome-wide association studies (GWAS) in Australia and England, there were no GWAS about Han Chinese to our knowledge. Thus, this research focused on using whole genomic genotypes from the Taiwan Biobank for exploring the association between Al concentrations in plasma and renal function. Participants, who underwent questionnaire interviews, biomarkers, and genotyping, were from the Taiwan Biobank database. Then, we measured their plasma Al concentrations with ICP-MS in the laboratory at Kaohsiung Medical University. We used this data to link genome-wide association (GWA) tests while looking for candidate genes and associated plasma Al concentration to renal function. Furthermore, we examined the path relationship between Single Nucleotide Polymorphisms (SNPs), Al concentrations, and estimated glomerular filtration rates (eGFR) through the mediation analysis with 3000 replication bootstraps. Following the principles of GWAS, we focused on three SNPs within the dipeptidyl peptidase-like protein 6 (DPP6) gene in chromosome 7, rs10224371, rs2316242, and rs10268004, respectively. The results of the mediation analysis showed that all of the selected SNPs have indirectly affected eGFR through a mediation of Al concentrations. Our analysis revealed the association between DPP6 SNPs, plasma Al concentrations, and eGFR. However, further longitudinal studies and research on mechanism are in need. Our analysis was still be the first study that explored the association between the DPP6, SNPs, and Al in plasma affecting eGFR.
Perceived Consequences of Extended Social Isolation on Mental Well-Being: Narratives from Indonesian University Students during the COVID-19 Pandemic
Authors: Rahiem Maila D. H. , Krauss Steven Eric , Ersing Robin
Published: October 2021

Despite several recent studies reporting on young people’s well-being during COVID-19, few large-scale qualitative studies have been carried out that capture the experiences of young people from low- and middle-income countries (LMICs) undergoing extended social restrictions. The challenges faced by young people from LMICs during COVID-19 are likely to be amplified by their countries’ large populations, resource constraints, lack of access to health care, living conditions, socio-spatial contexts, and the pandemic’s ramifications for communities. This study explored how youths perceived their well-being after being isolated for one-and-a-half years during the COVID-19 pandemic. Qualitative narrative research was employed as a method of inquiry. One-hundred and sixty-six university students in Jakarta, Indonesia, between the ages of 17 and 22 wrote reflective online essays on the consequences of extended pandemic isolation on their mental health. This data collection strategy offered an in-depth understanding of the phenomenon through the narratives of those who experienced it. Seven themes expressing the youths’ perceived well-being were identified through inductive reflective thematic analysis: (1) the anguish of loneliness and estrangement; (2) a state of “brokenness” resulting from emotional agony and distress; (3) frustration, confusion, and anger; (4) the experience of conflicting emotions; (5) uncertainty about both the present and future; (6) a sense of purpose and fulfillment; and (7) turning to faith. The findings provide important insights into Indonesian youths’ well-being following extended social restrictions following the outbreak. Their collective experiences can be used to inform policy and practice regarding the nature of support mechanisms required both during and following the pandemic, and in the future if such a situation were to occur again.
Utility of Self-Reported Heat Stress Symptoms and NGAL Biomarker to Screen for Chronic Kidney Disease of Unknown Origin (CKDu) in Sri Lanka
Authors: Kulasooriya Pavithra N. , Jayasekara Kithsiri B. , Nisansala Thilini , Kannangara Sajani , Karunarathna Ranawaka , Karunarathne Chaminda , Wikramarathne Mahinda , Albert Steven M.
Published: October 2021

Objective. We examined heat stress symptoms and urine markers of chronic kidney disease (CKDu) in Sri Lanka to assess differences between endemic vs. non-endemic regions and by occupation. Sample and Methods. We assessed a total of 475 villagers. In the endemic region, 293 were agricultural workers and 67 were not working primarily in agriculture. In the non-endemic region, 76 were agricultural workers. Of the residents, 218 were assessed for neutrophil gelatinase-associated lipocalin (NGAL), an early predictor of acute kidney injury, along with urine markers of chronic kidney disease. Results. The mean (sd) age of the sample was 45.2 (12.6), with males comprising 52.7%; 7.2% reported kidney disease (n = 34), and 5.7% reported diabetes (n = 27). The heat stress index (mean (sd)) was highest among agricultural workers in the endemic region (8.05 (5.9)), intermediate in non-agricultural workers in the endemic region (4.61 (4.5)), and lowest among agricultural workers in the non-endemic region (3.85 (3.3)); p < 0.0001. Correlations were higher between NGAL and serum microalbumin in the endemic agricultural worker sample than in the other two samples (Spearman’s r = 0.34 vs. 0.15 and 0.20). Conclusions. Both heat stress symptoms and NGAL values were higher among agricultural workers in endemic CKDu regions. Correlations between NGAL and microalbumin suggested a link between acute kidney injury and chronic kidney disease in the more-exposed sample.
Maternal Mortality from Induced Abortion in Malawi: What Does the Latest Evidence Suggest?
Authors: Miller Calum
Published: October 2021

It is commonly claimed that thousands of women die every year from unsafe abortion in Malawi. This commentary critically assesses those claims, demonstrating that these estimates are not supported by the evidence. On the contrary, the latest evidence—itself from 15 to 20 years ago—suggests that 6–7% of maternal deaths in Malawi are attributable to induced and spontaneous abortion combined, totalling approximately 70–150 deaths per year. I then offer some evidence suggesting that a substantial proportion of these are attributable to spontaneous abortion. To reduce maternal mortality by large margins, emergency obstetric care should be prioritised, which will also save women from complications of induced and spontaneous abortion.
The Demand–Control Model and Pupils’ Aggressive Behaviour towards Teachers: A Follow-Up Study
Authors: Andersen Lars Peter , Aust Birgit , Winding Trine Nøhr
Published: October 2021

Purpose: Pupils’ aggressive behaviour towards teachers is a common phenomenon in schools across different countries. The purpose of this study is to test hypotheses that are central to the Job Demand–Control model as risk factors for pupils’ aggressive behaviour towards teachers. Method: Questionnaire data were collected in 2018 and 2019 from teachers at 94 public schools in Denmark. In total, 1198 teachers participated in both rounds. Demands and social support at work were measured in 2018, and pupils’ aggressive behaviour was measured in 2019. The analyses were performed using multivariate logistic regression analysis. Results: Teachers were often exposed to pupils’ aggressive behaviour during their work. High emotional work demands and low control were associated with increased risk of pupils’ aggressive behaviour. No mitigating effect of high control was found on the association between emotional demands and risk for pupils’ aggressive behaviour towards teachers. Conclusion: High emotional demands were strongly associated with the aggressive behaviour of pupils towards teachers. Job control over own work situation was not enough to lower the risk of aggressive behaviour under conditions in which teachers experience high emotional demands. Based on these results, we recommend that supervisors carefully balance teachers’ emotional demands to their resources.
Overview of Human HtrA Family Proteases and Their Distinctive Physiological Roles and Unique Involvement in Diseases, Especially Cancer and Pregnancy Complications
Authors: Wang Yao , Nie Guiying
Published: October 2021

The mammalian high temperature requirement A (HtrA) proteins are a family of evolutionarily conserved serine proteases, consisting of four homologs (HtrA1-4) that are involved in many cellular processes such as growth, unfolded protein stress response and programmed cell death. In humans, while HtrA1, 2 and 3 are widely expressed in multiple tissues with variable levels, HtrA4 expression is largely restricted to the placenta with the protein released into maternal circulation during pregnancy. This limited expression sets HtrA4 apart from the rest of the family. All four HtrAs are active proteases, and their specific cellular and physiological roles depend on tissue type. The dysregulation of HtrAs has been implicated in many human diseases such as cancer, arthritis, neurogenerative ailments and reproductive disorders. This review first discusses HtrAs broadly and then focuses on the current knowledge of key molecular characteristics of individual human HtrAs, their similarities and differences and their reported physiological functions. HtrAs in other species are also briefly mentioned in the context of understanding the human HtrAs. It then reviews the distinctive involvement of each HtrA in various human diseases, especially cancer and pregnancy complications. It is noteworthy that HtrA4 expression has not yet been reported in any primary tumour samples, suggesting an unlikely involvement of this HtrA in cancer. Collectively, we accentuate that a better understanding of tissue-specific regulation and distinctive physiological and pathological roles of each HtrA will improve our knowledge of many processes that are critical for human health.
Auxin Treatment Enhances Anthocyanin Production in the Non-Climacteric Sweet Cherry (Prunus avium L.)
Authors: Clayton-Cuch Daniel , Yu Long , Shirley Neil , Bradley David , Bulone Vincent , Böttcher Christine
Published: October 2021

Abscisic acid (ABA) is a key signaling molecule promoting ripening of non-climacteric fruits such as sweet cherry (Prunus avium L.). To shed light on the role of other hormones on fruit development, ripening and anthocyanin production, the synthetic auxin 1-naphthaleneacetic acid (NAA) was applied to sweet cherry trees during the straw-color stage of fruit development. NAA-treated fruits exhibited higher concentrations of 1-aminocyclopropane-1-carboxylic acid (ACC) and ABA-glucose ester (ABA-GE), which are a precursor of ethylene and a primary storage form of ABA, respectively. Consistent with these observations, transcript levels of genes encoding ACC synthase and ACC oxidase, both involved in ethylene biosynthesis, were increased after 6 days of NAA treatment, and both ABA concentration and expression of the regulator gene of ABA biosynthesis (NCED1 encoding 9-cis-epoxycarotenoid dioxygenase) were highest during early fruit ripening. In addition, transcript levels of key anthocyanin regulatory, biosynthetic and transport genes were significantly upregulated upon fruit exposure to NAA. This was accompanied by an increased anthocyanin concentration and fruit weight whilst fruit firmness and cracking index decreased. Altogether our data suggest that NAA treatment alters ethylene production, which in turn induces ripening in sweet cherry and enhanced anthocyanin production, possibly through ABA metabolism. The results from our study highlight the potential to use a single NAA treatment for manipulation of cherry ripening.
Dye-Mediated Photo-Oxidation Biomaterial Fixation: Analysis of Bioinductivity and Mechanical Properties of Bovine Pericardium for Use in Cardiac Surgery
Authors: Pattar Simranjit S. , Vasanthan Vishnu , Teng Guoqi , Wagner Karl T. , Jeon Kristina , Kang Sean , Fatehi Hassanabad Ali , Fedak Paul W. M.
Published: October 2021

Extracellular matrix bioscaffolds can influence the cardiac microenvironment and modulate endogenous cellular mechanisms. These materials can optimize cardiac surgery for repair and reconstruction. We investigated the biocompatibility and bioinductivity of bovine pericardium fixed via dye-mediated photo-oxidation on human cardiac fibroblast activity. We compared a dye-mediated photo-oxidation fixed bioscaffold to glutaraldehyde-fixed and non-fixed bioscaffolds reported in contemporary literature in cardiac surgery. Human cardiac fibroblasts from consenting patients were seeded on to bioscaffold materials to assess the biocompatibility and bioinductivity. Human cardiac fibroblast gene expression, secretome, morphology and viability were studied. Dye-mediated photo-oxidation fixed acellular bovine pericardium preserves human cardiac fibroblast phenotype and viability; and potentiates a pro-vasculogenic paracrine response. Material tensile properties were compared with biomechanical testing. Dye-mediated photo-oxidation fixed acellular bovine pericardium had higher compliance compared to glutaraldehyde-fixed bioscaffold in response to tensile force. The biocompatibility, bioinductivity, and biomechanical properties of dye-mediated photo-oxidation fixed bovine pericardium demonstrate its feasibility as a bioscaffold for use in cardiac surgery. As a fixed yet bioinductive solution, this bioscaffold demonstrates enhanced compliance and retains bioinductive properties that may leverage endogenous reparative pathways. Dye-mediated photo-oxidation fixed bioscaffold warrants further investigation as a viable tool for cardiac repair and reconstruction.
Impact of the Pd2Spm (Spermine) Complex on the Metabolism of Triple-Negative Breast Cancer Tumors of a Xenograft Mouse Model
Authors: Carneiro Tatiana J. , Araújo Rita , Vojtek Martin , Gonçalves-Monteiro Salomé , de Carvalho Ana L. M. Batista , Marques Maria Paula M. , Diniz Carmen , Gil Ana M.
Published: October 2021

The interest in palladium(II) compounds as potential new anticancer drugs has increased in recent years, due to their high toxicity and acquired resistance to platinum(II)-derived agents, namely cisplatin. In fact, palladium complexes with biogenic polyamines (e.g., spermine, Pd2Spm) have been known to display favorable antineoplastic properties against distinct human breast cancer cell lines. This study describes the in vivo response of triple-negative breast cancer (TNBC) tumors to the Pd2Spm complex or to cisplatin (reference drug), compared to tumors in vehicle-treated mice. Both polar and lipophilic extracts of tumors, excised from a MDA-MB-231 cell-derived xenograft mouse model, were characterized through nuclear magnetic resonance (NMR) metabolomics. Interestingly, the results show that polar and lipophilic metabolomes clearly exhibit distinct responses for each drug, with polar metabolites showing a stronger impact of the Pd(II)-complex compared to cisplatin, whereas neither drug was observed to significantly affect tumor lipophilic metabolism. Compared to cisplatin, exposure to Pd2Spm triggered a higher number of, and more marked, variations in some amino acids, nucleotides and derivatives, membrane precursors (choline and phosphoethanolamine), dimethylamine, fumarate and guanidine acetate, a signature that may be relatable to the cytotoxicity and/or mechanism of action of the palladium complex. Putative explanatory biochemical hypotheses are advanced on the role of the new Pd2Spm complex in TNBC metabolism.
PARN-like Proteins Regulate Gene Expression in Land Plant Mitochondria by Modulating mRNA Polyadenylation
Authors: Hirayama Takashi
Published: October 2021

Mitochondria have their own double-stranded DNA genomes and systems to regulate transcription, mRNA processing, and translation. These systems differ from those operating in the host cell, and among eukaryotes. In recent decades, studies have revealed several plant-specific features of mitochondrial gene regulation. The polyadenylation status of mRNA is critical for its stability and translation in mitochondria. In this short review, I focus on recent advances in understanding the mechanisms regulating mRNA polyadenylation in plant mitochondria, including the role of poly(A)-specific ribonuclease-like proteins (PARNs). Accumulating evidence suggests that plant mitochondria have unique regulatory systems for mRNA poly(A) status and that PARNs play pivotal roles in these systems.
Human Liver Stem Cell Derived Extracellular Vesicles Alleviate Kidney Fibrosis by Interfering with the β-Catenin Pathway through miR29b
Authors: Kholia Sharad , Herrera Sanchez Maria Beatriz , Deregibus Maria Chiara , Sassoè-Pognetto Marco , Camussi Giovanni , Brizzi Maria Felice
Published: October 2021

Human liver stem-cell-derived extracellular vesicles (HLSC-EVs) exhibit therapeutic properties in various pre-clinical models of kidney injury. We previously reported an overall improvement in kidney function following treatment with HLSC-EVs in a model of aristolochic acid nephropathy (AAN). Here, we provide evidence that HLSC-EVs exert anti-fibrotic effects by interfering with β-catenin signalling. A mouse model of AAN and an in vitro pro-fibrotic model were used. The β-catenin mRNA and protein expression, together with the pro-fibrotic markers α-SMA and collagen 1, were evaluated in vivo and in vitro following treatment with HLSC-EVs. Expression and functional analysis of miR29b was performed in vitro following HLSC-EV treatments through loss-of-function experiments. Results showed that expression of β-catenin was amplified both in vivo and in vitro, and β-catenin gene silencing in fibroblasts prevented AA-induced up-regulation of pro-fibrotic genes, revealing that β-catenin is an important factor in fibroblast activation. Treatment with HLSC-EVs caused increased expression of miR29b, which was significantly inhibited in the presence of α-amanitin. The suppression of the miR29b function with a selective inhibitor abolished the anti-fibrotic effects of HLSC-EVs, resulting in the up-regulation of β-catenin and pro-fibrotic α-Sma and collagen type 1 genes. Together, these data suggest a novel HLSC-EV-dependent regulatory mechanism in which β-catenin is down regulated by HLSC-EVs-induced miR29b expression.
Cellular and Molecular Targets of Nucleotide-Tagged Trithiolato-Bridged Arene Ruthenium Complexes in the Protozoan Parasites Toxoplasma gondii and Trypanosoma brucei
Authors: Anghel Nicoleta , Müller Joachim , Serricchio Mauro , Jelk Jennifer , Bütikofer Peter , Boubaker Ghalia , Imhof Dennis , Ramseier Jessica , Desiatkina Oksana , Păunescu Emilia , Braga-Lagache Sophie , Heller Manfred , Furrer Julien , Hemphill Andrew
Published: October 2021

Toxoplasma gondii is an apicomplexan parasite that infects and proliferates within many different types of host cells and infects virtually all warm-blooded animals and humans. Trypanosoma brucei is an extracellular kinetoplastid that causes human African trypanosomiasis and Nagana disease in cattle, primarily in rural sub-Saharan Africa. Current treatments against both parasites have limitations, e.g., suboptimal efficacy and adverse side effects. Here, we investigate the potential cellular and molecular targets of a trithiolato-bridged arene ruthenium complex conjugated to 9-(2-hydroxyethyl)-adenine (1), which inhibits both parasites with IC50s below 10−7 M. Proteins that bind to 1 were identified using differential affinity chromatography (DAC) followed by shotgun-mass spectrometry. A trithiolato-bridged ruthenium complex decorated with hypoxanthine (2) and 2-hydroxyethyl-adenine (3) were included as controls. Transmission electron microscopy (TEM) revealed distinct ultrastructural modifications in the mitochondrion induced by (1) but not by (2) and (3) in both species. DAC revealed 128 proteins in T. gondii and 46 proteins in T. brucei specifically binding to 1 but not 2 or 3. In T. gondii, the most abundant was a protein with unknown function annotated as YOU2. This protein is a homolog to the human mitochondrial inner membrane translocase subunit Tim10. In T. brucei, the most abundant proteins binding specifically to 1 were mitochondrial ATP-synthase subunits. Exposure of T. brucei bloodstream forms to 1 resulted in rapid breakdown of the ATP-synthase complex. Moreover, both datasets contained proteins involved in key steps of metabolism and nucleic acid binding proteins.
Exosite Binding in Thrombin: A Global Structural/Dynamic Overview of Complexes with Aptamers and Other Ligands
Authors: Troisi Romualdo , Balasco Nicole , Autiero Ida , Vitagliano Luigi , Sica Filomena
Published: October 2021

Thrombin is the key enzyme of the entire hemostatic process since it is able to exert both procoagulant and anticoagulant functions; therefore, it represents an attractive target for the developments of biomolecules with therapeutic potential. Thrombin can perform its many functional activities because of its ability to recognize a wide variety of substrates, inhibitors, and cofactors. These molecules frequently are bound to positively charged regions on the surface of protein called exosites. In this review, we carried out extensive analyses of the structural determinants of thrombin partnerships by surveying literature data as well as the structural content of the Protein Data Bank (PDB). In particular, we used the information collected on functional, natural, and synthetic molecular ligands to define the anatomy of the exosites and to quantify the interface area between thrombin and exosite ligands. In this framework, we reviewed in detail the specificity of thrombin binding to aptamers, a class of compounds with intriguing pharmaceutical properties. Although these compounds anchor to protein using conservative patterns on its surface, the present analysis highlights some interesting peculiarities. Moreover, the impact of thrombin binding aptamers in the elucidation of the cross-talk between the two distant exosites is illustrated. Collectively, the data and the work here reviewed may provide insights into the design of novel thrombin inhibitors.
Residual Helicity at the Active Site of the Histidine Phosphocarrier, HPr, Modulates Binding Affinity to Its Natural Partners
Authors: Neira José L. , Ortega-Alarcón David , Rizzuti Bruno , Palomino-Schätzlein Martina , Velázquez-Campoy Adrián , Falcó Alberto
Published: October 2021

The phosphoenolpyruvate-dependent phosphotransferase system (PTS) modulates the preferential use of sugars in bacteria. The first proteins in the cascade are common to all organisms (EI and HPr). The active site of HPr involves a histidine (His15) located immediately before the beginning of the first α-helix. The regulator of sigma D (Rsd) protein also binds to HPr. The region of HPr comprising residues Gly9-Ala30 (HPr9–30), involving the first α-helix (Ala16-Thr27) and the preceding active site loop, binds to both the N-terminal region of EI and intact Rsd. HPr9–30 is mainly disordered. We attempted to improve the affinity of HPr9–30 to both proteins by mutating its sequence to increase its helicity. We designed peptides that led to a marginally larger population in solution of the helical structure of HPr9–30. Molecular simulations also suggested a modest increment in the helical population of mutants, when compared to the wild-type. The mutants, however, were bound with a less favorable affinity than the wild-type to both the N-terminal of EI (EIN) or Rsd, as tested by isothermal titration calorimetry and fluorescence. Furthermore, mutants showed lower antibacterial properties against Staphylococcus aureus than the wild-type peptide. Therefore, we concluded that in HPr, a compromise between binding to its partners and residual structure at the active site must exist to carry out its function.
CNPase, a 2′,3′-Cyclic-nucleotide 3′-phosphodiesterase, as a Therapeutic Target to Attenuate Cardiac Hypertrophy by Enhancing Mitochondrial Energy Production
Authors: Tan Keai Sinn , Wang Dongfang , Lu Ziqiang , Zhang Yihan , Li Sixu , Lin Yue , Tan Wen
Published: October 2021

Heart failure is the end-stage of all cardiovascular diseases with a ~25% 5-year survival rate, and insufficient mitochondrial energy production to meet myocardial demand is the hallmark of heart failure. Mitochondrial components involved in the regulation of ATP production remain to be fully elucidated. Recently, roles of 2′,3′-cyclic nucleotide-3′-phosphodiesterase (CNPase) in the pathophysiological processes of heart diseases have emerged, implicated by evidence that mitochondrial CNPase proteins are associated with mitochondrial integrity under metabolic stress. In this study, a zebrafish heart failure model was established, by employing antisense morpholino oligonucleotides and the CRISPR-Cas9 gene-editing system, which recapitulates heart failure phenotypes including heart dysfunction, pericardial edema, ventricular enlargement, bradycardia, and premature death. The translational implications of CNPase in the pathophysiological process of heart failure were tested in a pressure overload-induced heart hypertrophy model, which was carried out in rats through transverse abdominal aorta constriction (TAAC). AAV9-mediated myocardial delivery of CNPase mitigated the hypertrophic response through the specific hydrolysis of 2′-3′-cyclic nucleotides, supported by the decrease of cardiac hypertrophy and fibrosis, the integrity of mitochondrial ultrastructure, and indicators of heart contractility in the AAV9-TAAC group. Finally, the biometrics of a mitochondrial respiration assay carried out on a Seahorse cellular energy analyzer demonstrated that CNPase protects mitochondrial respiration and ATP production from AngII-induced metabolic stress. In summary, this study provides mechanistic insights into CNPase-2′,3′-cyclic nucleotide metabolism that protects the heart from energy starvation and suggests novel therapeutic approaches to treat heart failure by targeting CNPase activity.