bluuebunny/crossref_metadata_2025 huggingface dataset:
Numerical Analysis of Flow Boiling Characteristics of a Single Channel Heat Sink Subjected to Multiple Heat Sources
Authors: Qian Jiyu , Wang Rui , Wei Tao , Tang Hao , Hu Dinghua
Published: March 2023

High-power electronic devices with multiple heat sources often require temperature uniformity and to operate within their functional temperature range for optimal performance. Micro-channel cooling could satisfy the heat dissipation requirements, but it may cause temperature non-uniformity. In this paper, simulations are performed for different geometric parameters of the channel and the position of the heat source. The results show that a flattened channel can effectively reduce the heat source temperature, broadening the straight channel can reduce the flow resistance and enhance heat transfer, while widening the channel at the bend may lead to local dryness. Meanwhile, a thermal model is established to analyze the influence of the position of the heat source. The results also show that with the increase in the curved channel radius, the phenomenon of vapor–liquid separation becomes more obvious, the pressure drop decreases, but the heat transfer effect worsens.
Influence of Temperature in the Thermo-Chemical Decomposition of Below-Stoichiometric RDF Char—A Macro TGA Study
Authors: Castro Carlos , Gonçalves Margarida , Longo Andrei , Vilarinho Cândida , Ferreira Manuel , Ribeiro André , Pacheco Nuno , Teixeira José C.
Published: March 2023

Due to the energy crisis that some countries are facing nowadays, the gasification process appears to be a good alternative to produce some energy from solid materials. Increasingly, gasification involves using wastes as a solid fuel, making the process green and reusing some materials that otherwise could end up in a landfill. However, the process of finding the best gasification parameters of a sample can be very expensive and time-consuming. In this sense, a refuse-derived fuel (RDF) char produced from an original RDF under 30 min at 400 °C was tested on a small-scale reactor using macro thermogravimetric analysis (TGA), as presented in this paper. The goal was to study and evaluate the devolatilization and residual carbon rate of the sample under several conditions and, at the same time, quantify and analyze the released gas. In the first round of tests, 5, 10, and 20 g of samples were tested at 750 °C with an excess of air coefficient (λ) = 0 and 0.2. It was possible to conclude that the lower the mass, the higher the devolatilization rate. The λ only had an influence on the devolatilization rate with a 20 g sample. Regarding the gas, CO, CO2, and H2 had no variation in the sample mass in contrast to CH4, which increased with the increase in the sample mass. The second round of tests was performed with samples of 10 g of mass at temperatures of 700, 800, and 900 °C and λ values of 0.15, 0.2, and 0.25. The tests indicated that the temperature influenced the devolatilization rate but not the residual carbon combustion rate. Regarding the gas composition, CH4, CO2, and CO followed the same trend, decreasing the concentration with the increase in temperature; in contrast, H2 increased in concentration with an increase in temperature. The heating value of the gas followed the same behavior as CH4.
Towards an Inclusive Approach to Forest Management: Highlight of the Perception and Participation of Local Communities in the Management of miombo Woodlands around Lubumbashi (Haut-Katanga, D.R. Congo)
Authors: N’tambwe Nghonda Dieu-donné , Muteya Héritier Khoji , Kashiki Bill Kasongo Wa Ngoy , Sambiéni Kouagou Raoul , Malaisse François , Sikuzani Yannick Useni , Kalenga Wilfried Masengo , Bogaert Jan
Published: March 2023

The misappropriation of sustainable forest programs by local communities and the under-utilization of their knowledge are major impediments to the mitigation of deforestation. Within this context, participation has become a principle used in almost all interventions. It is important to evaluate the practices in this area to ensure better involvement of local communities. This survey examined the perception and participation of local communities in the management of miombo woodlands, based on semi-structured questionnaire surveys involving 945 households in 5 villages in the Lubumbashi rural area. The results reveal that local communities perceive soil fertility loss and deforestation as major environmental challenges in their area. This perception remains largely influenced by their socio-demographic factors such as respondents’ age, seniority in the villages, and level of education. To mitigate deforestation, the rare actions of provincial public services and non-governmental organizations are focused on the sustainable exploitation of miombo woodlands through the development of simple management plans, reforestation, and forest control. These activities are sparse and poorly inclusive of scientific findings and the priorities of local communities. These justify poor community participation, particularly in the actions of provincial public services. For a better appropriation of sustainable forest management plans and to reinforce miombo woodlands’ resilience to anthropogenic pressures, based on these findings, we recommend a concerted and inclusive approach to forest planning.
Unlocking Digitalization in Forest Operations with Viewshed Analysis to Improve GNSS Positioning Accuracy
Authors: Lopatin Eugene , Väätäinen Kari , Kukko Antero , Kaartinen Harri , Hyyppä Juha , Holmström Eero , Sikanen Lauri , Nuutinen Yrjö , Routa Johanna
Published: March 2023

This study evaluated the positioning accuracy of moving forest harvesters using global navigation satellite system (GNSS) signals under a forest canopy, and developed approaches for forecasting accuracy under a mature spruce canopy cover. Real-time kinematic positioning with a Trimble R12 receiver on top of a harvester achieved high positioning accuracy, with 86% of observations meeting a maximum precision of 8 mm. However, the presence of a canopy cover hampered the GNSS’s performance as there were fewer satellites available, leading to an increased number of inaccurate positions and larger values of the dilution of precision in geometry (GDOP), position (PDOP), vertical (VDOP) and horizontal directions (HDOP). The canopy cover estimated from the viewshed analysis of the digital surface model (DSM) was found to be a significant predictor of the dilution of precision and maximum deviation from the true position. These findings suggest that viewshed analysis provides more precise results than a mere canopy cover percentage for evaluating the impact of canopy cover on the GNSS’s positioning of a harvester, despite its computational demands. Developing intelligent algorithms for precise positioning under the canopy can facilitate autonomous harvesting and forwarding, allowing for the implementation of digitalization in forest operations.
Niche Dynamics Below the Species Level: Evidence from Evaluating Niche Shifts within Quercus aquifolioides
Authors: Feng Li , Zhou Lipan , Zhang Tianyi , Wang Xumei
Published: March 2023

The role of ecological niches in lineage diversification has been the subject of long-standing interest of ecologists and evolutionary biologists. Specific responses to climate change can arise below the species level, resulting in differentiated adaptation or movement patterns within a given species. Thus, the urgent need to model potential responses to ongoing climate changes among genetically distinct populations within a species is increasingly recognized. In this study, we utilize the information of intraspecific variation within Quercus aquifolioides as a priori, and then focus on the potential distribution ranges and niche dynamics of its three intraspecific lineages (WSP, HDM, and Tibet) under current environmental conditions via ecological niche models (ENMs) and PCA-env ordination method, respectively. Our results indicated that the three lineages have occupied differentiated climatic niches. Although the three lineages have distinctly adaptive strategies for homogeneous environmental conditions, some lineages had sympatric projecting areas. The PCA-env demonstrated that the lineage pair WSP vs. HDM had the largest niche overlap while Tibet vs. HDM showed the smallest one. Moreover, the hypothesis of the niche was indistinguishable within the three lineages rejected, indicating the presence of niche divergence rather than niche conservatism below the species level. Our findings highlight the potential of modeling intraspecific responses to climate change and provide insights into lineage diversification within Q. aquifolioides, permitting the exploration of the information determined by niche evaluations and comparisons to understand plant diversification processes below the species level in biodiversity hotspots.
Effect of Elevated Temperature on the Behavior of Amorphous Metallic Fibre-Reinforced Cement and Geopolymer Composites
Authors: Shaikh Faiz Uddin Ahmed , Kahlon Narwinder Singh , Dogar Attiq Ur Rahman
Published: March 2023

To improve the tensile, flexural, and ductility properties of geopolymer composites, amorphous metallic fibres (AMF) are used to reinforce these composites, and the behavior of these composites at elevated temperatures has been assessed in this study. Four types of composites, i.e., cement, reinforced cement, geopolymer, and reinforced geopolymer composites have been prepared. The composites have been reinforced using AMF with a fibre volume fraction of 0.75%. The composites have been assessed for change in mass loss, cracking, compressive strength, and flexural strength at four elevated temperatures of 200 °C, 400 °C, 600 °C, and 800 °C, and conclusions have been drawn concerning these composites. The results have shown that an increase in temperature has an adverse effect on these composites, and geopolymer composites exhibit higher performance than their counterpart cement composites at elevated temperatures. The mass loss and surface cracking were significantly lower in geopolymer composites, and the fibre reinforcement had a negligible effect on mass loss. Also, the residual compressive and flexural strength of reinforced geopolymer composites was significantly higher than that of the reinforced cement composites. In addition, scanning electron microscopic images also showed that even at higher temperatures, the geopolymer matrix is present on the AMF fibre, which results in higher residual strength than the cement composites in which a negligible amount of matrix is present on the fibres.
Did Household Income Loss Have an Immediate Impact on Animal-Source Foods Consumption during the Early Stage of the COVID-19 Pandemic?
Authors: Shen Qi , Zhong Taiyang
Published: March 2023

The outbreak of COVID-19 in 2020 caused extensive impact on household income and foods consumption. However, little attention has been paid to the immediate impact of income loss on animal-source foods consumption in the early stage of the COVID-19 pandemic. This paper aims to narrow this gap, and a total of 1301 valid samples of household food consumption surveys in Wuhan and Nanjing were obtained through specially designed online questionnaires. The surveys show that there were 69.6% (Wuhan) and 42.2% (Nanjing) of surveyed households whose animal-source foods consumption were affected, and there were 47.4% (Wuhan) and 18.9% (Nanjing) of surveyed households who suffered income loss. Furthermore, this paper makes an empirical study on the linkage between income loss and animal-source foods consumption. The results show that the pandemic affected household income, resulting in an immediate impact on animal-source foods consumption. This immediate impact may have been due to the combination of price increases, income loss and insufficient savings, which led to a “perfect storm” for animal-source foods consumption. Moreover, household income loss affected various animal-source foods consumption differently. For households suffering income losses, the odds of pork, beef and mutton, poultry, aquatic products, eggs and dairy products consumption being affected were increased by a factor of 1.894, 2.140, 2.773, 2.345, 1.802, 2.835, respectively, holding other variables constant. The results may be related to residents’ consumption habits and food prices. During the COVID-19 pandemic, the reduction of animal-source foods consumption may have led to a state of tension concerning an increase in the development of nutrition intake and health, which may have led to increased food security risks.
Physicochemical Attributes, Volatile Profile and Sensory Quality of Organic Crimson Crisp Apples during On-Tree Maturation
Authors: Pontesegger Niklas , Rühmer Thomas , Siegmund Barbara
Published: March 2023

When new apple cultivars are planted, knowledge of their maturation and ripening behavior and, as a consequence, the best time for their harvest is of utmost importance for providing fruits of the highest quality to consumers. In this study, we followed the on-tree maturation and ripening behavior of organic Crimson Crisp apples over a period of almost 3 months. With the weekly analyses of basic fruit quality attributes (weight, fruit firmness, total soluble solids, titratable acids, starch degradation) in combination with the fruit volatilome (primary and secondary volatiles) and sensory analysis, we obtained a holistic picture of the maturation and ripening properties of this new variety. We could show that at the recommended harvest window, which is based on the degree of starch degradation alone, the fruit development is not finished. Synthesis of aroma volatiles—which is strongly related to the expression of pronounced fruity, apple-like flavor—requires two additional weeks of on-tree ripening. Results indicate an upregulation of 13-LOX at very early maturation stages, while upregulation of 9-LOX and enzyme systems involved in the β-oxidation pathway requires a prolonged on-tree maturation period. The results of this study demonstrate that the analysis of basic fruit quality attributes is not sufficient for understanding the properties of apples. However, we demonstrate that the analysis of the fruit volatilome is a valuable and necessary tool for optimizing the quality of new apple varieties.
Japanese Translation and Validation of Genomic Knowledge Measure in the International Genetics Literacy and Attitudes Survey (iGLAS-GK)
Authors: Yoshida Akiko , Tokutomi Tomoharu , Fukushima Akimune , Chapman Robert , Selita Fatos , Kovas Yulia , Sasaki Makoto
Published: March 2023

Knowledge of genetics is essential for understanding the results of genetic testing and its implications. Recent advances in genomic research have allowed us to predict the risk of onset of common diseases based on individual genomic information. It is anticipated that more people will receive such estimates of risks based on their genomic data. However, currently, there is no measure for genetic knowledge that includes post-genome sequencing advancements in Japan. In this study, we translated the genomic knowledge measure in the International Genetics Literacy and Attitudes Survey (iGLAS-GK) into Japanese and validated it in a general Japanese adult population (n = 463). The mean score was 8.41 (SD 2.56, range 3–17). The skewness and kurtosis were 0.534 and 0.088, respectively, and the distribution showed a slightly positive skewness. Exploratory factor analysis proposed a six-factor model. Results for 16 of the 20 items of the Japanese version of the iGLAS-GK were comparable to those from previous studies in other populations. These results indicate that the Japanese version is reliable and can be used to measure the genomic knowledge of adults in the general population, and this version of the knowledge measure maintains the multidimensional structure for assessing genomic knowledge.
Neurological Implications of Vitamin B12 Deficiency in Diet: A Systematic Review and Meta-Analysis
Authors: Alruwaili Mubarak , Basri Rehana , AlRuwaili Raed , Albarrak Anas Mohammad , Ali Naif H.
Published: March 2023

Background: Vitamin B12 is one of the most important B-Vitamins that the human body needs on a daily basis, the lack of which can precipitate several neurological issues. Objectives: This systematic aimed to investigate the neurological implications of Vitamin B12 deficiency and the effects when B12 levels were corrected in susceptible individuals. Methods: The databases PubMed-MEDLINE, Web of Science, Cochrane, and Scopus were all searched using pertinent keywords, reference searches, and citation searches. The terms used to access the database were “Cognition”, “Dietary patterns”, “Neurology”, “Nutritional profile”, and “Vitamin B12”. Results: Vitamin B12 was shown to noticeably improve cognition and other neurological parameters in the short term in older adults and the short-to-medium term in children; however, there was no perceived increase/improvement when the Vitamin was administered in the longer term, either alone or in conjunction with other similar nutritional interventions. Conclusion: Vitamin B12’s role in the improvement of neurological functions over a long-term period remains somewhat inconclusive to date, as the majority of our selected control trials did not display much correlation between the two factors. However, Vitamin B12 did improve cognition levels in both children and older adults over a short course of administration.
Fibromyalgia Rapid Screening Tool (FiRST): Arabic Translation and Cross-Cultural Adaptation and Validation
Authors: AlAujan Shiekha S. , Almalag Haya M. , Assiri Ghadah A. , Alodaibi Faris A. , Omair Mohammed A.
Published: March 2023

Background: Fibromyalgia (FM), a complex neurological disorder, has multiple consequences for the patient. To diagnose patients, healthcare practitioners use multiple diagnostic questionnaires. However, Arabic translated or validated tools are lacking. This study aimed to translate and validate the Fibromyalgia Rapid Screening Tool (FiRST) into the Arabic language. Methods: Forward and backward translations of the FiRST were conducted by two Arabic translators and two English-certified translators. The survey was piloted (n = 5) and subjected to cognitive interviews and psychometric analysis. Patients were recruited from a university hospital in Riyadh and an FM support group in Saudi Arabia. The internal consistency, factor analysis, and test–retest correlations were evaluated. Results: This study included 46 patients. The stepwise translation process resulted in minor edits related to the use of synonyms to the survey items. The translated survey had a good internal consistency and test–retest correlation, with a Cronbach’s alpha of 0.7 and Pearson’s correlation coefficient of 0.79 (p-value < 0.001), respectively. The survey was factorable into two themes: generalized symptoms and more specific sensations. Conclusions: The Arabic FiRST is a simple, valid, and reliable tool to diagnose patients with FM in different settings.
The Association between the Atherogenic Index of Plasma and Cardiometabolic Risk Factors: A Review
Authors: Lioy Beatrice , Webb Richard James , Amirabdollahian Farzad
Published: March 2023

Background: Metabolic syndrome (MetS) is a condition caused by a combination of cardiometabolic risk factors (CMR). MetS leads to type 2 diabetes mellitus (T2DM) and cardiovascular disease (CVD), both of which place a burden on not only the patients but also the healthcare system. Diagnostic criteria for MetS vary, and there is no universal tool to detect it. Recently, many studies have found positive associations between the atherogenic index of plasma (AIP) and some CMR factors. Therefore, a comprehensive review was needed to recapitulate these studies and qualitatively estimate the likelihood of AIP being associated with CMR. We aimed to review and summarise observational data on AIP and CMR factors and verify their association. Materials and Methods: A review of observational studies was conducted by searching “atherogenic index of plasma” in PubMed, One Search, and the Cochrane library. A total of 2068 articles were screened, and 32 were included after excluding paediatric, non-human and interventional studies, and those carried out on cohorts with conditions unrelated to MetS or on lipid-lowering medication. The Newcastle-Ottawa scale was used to assess their quality. Results: Most studies that reported high waist circumference (WC), triglycerides (TG), insulin resistance (IR) and low high-density lipoprotein cholesterol (HDL-C) concentration, also reported high AIP. Few studies investigated blood pressure (BP) and some discrepancies existed between their results. Conclusion: AIP may be associated with WC, TG, IR, and HDL-C. It is unclear if AIP is associated with BP. The current study’s results should be used to inform futureward a meta-analysis to be seen quantitatively. It is also recommended that more cohort studies stratified by gender and ethnicity be performed to ascertain if AIP can predict MetS before it manifests.
A Meta-Analysis and Systematic Review of Community-Based Intimate Partner Violence Interventions in India
Authors: Mittal Mona , Paden McCormick Anna , Palit Manjushree , Trabold Nicole , Spencer Chelsea
Published: March 2023

Intimate partner violence (IPV) in India remains an entrenched and prevalent public health issue. Despite ample evidence of the widespread problem of IPV in India and associated mental and physical morbidities, far less is known about intervention models to reduce IPV in India. The aims of this meta-analysis and systematic review are to assess the effectiveness of community-based interventions to reduce IPV in India and to provide a narrative synthesis of these intervention approaches. A total of 9 databases were searched to identify peer-reviewed, English-language articles published between January 2000 and September 2022. The search identified 10 studies that met study inclusion criteria, including 3 randomized control trials, 4 quasi-experimental, 2 pre/post, and 1 time-series evaluation. Eight studies were included in the meta-analysis. There was notable variation in the interventions and approaches employed to reduce IPV and varying measurement of IPV outcomes. The results of the meta-analysis show that participating in community-based IPV interventions produced a significant reduction in IPV among women. When considering different types of IPV, study participants were less likely to report physical and psychological IPV victimization. In addition, participants were also less likely to report approving of IPV after participating in community-based IPV interventions. Community-based interventions and research addressing IPV are still evolving in India. Missing descriptions of theoretical frameworks, sampling, intervention design, and inadequately reported effectiveness of intervention (both quantitative and qualitative reporting) need to be addressed. Moreover, long-term evaluations of the pilot interventions are needed to provide a clear picture of the long-term effectiveness, sustainability, and replicability of the community-based IPV interventions. The findings have implications for researchers, practitioners (community health workers, clinicians, and social workers), and policymakers keen on IPV reduction in India and globally.
Effects of Cognitive Appraisals on Perceived Self-Efficacy and Distress during the COVID-19 Lockdown: An Empirical Analysis Based on Structural Equation Modeling
Authors: Diotaiuti Pierluigi , Valente Giuseppe , Mancone Stefania , Corrado Stefano , Bellizzi Fernando , Falese Lavinia , Langiano Elisa , Vilarino Guilherme Torres , Andrade Alexandro
Published: March 2023

During the COVID-19 lockdown, individuals and households had to responsibly manage the difficulties and problems caused by the restrictions on their mobility, such as the interruptions to work schedules, insecure food supplies, and the procurement of services and health care. The perceptions of risk as well as the fear of disease were strongly linked to worry, defined as a sequence of thoughts that evoke negative emotions and elevated levels of anxiety and distress. This study evaluated how different cognitive appraisals of an individual’s sources of worry could influence their perceived self-efficacy and directly or indirectly moderate their perceived general distress. A sample of 544 participants completed a survey that included questions based on the generalized self-efficacy scale, the sources of concern during the epidemic, the cognitive appraisal scale in emergency, and perceived discomfort. Subsequently, a structural-equation-modeling (SEM) analysis was performed to test the moderating role of cognitive appraisals and perceived self-efficacy on participants’ overall distress during a lockdown. Overall, the model reported acceptable fit values and confirmed the hypotheses of the study. An individual’s worries activated either a dysfunctional threat appraisal, which prompted a fear-and-closure response that then increased their overall state of distress; or two other functional appraisals (i.e., adaptive or supportive) that contributed to significantly improving the individual’s self-efficacy. Higher self-efficacy was shown to be associated with lower levels of perceived distress. For the purposes of prevention and distress containment, it would be appropriate to promote psycho-educational interventions that promote the adoption of appraisal strategies that are functional and beneficial for perceived self-efficacy.
CsCIPK11-Regulated Metalloprotease CsFtsH5 Mediates the Cold Response of Tea Plants
Authors: Di Taimei , Wu Yedie , Peng Jing , Wang Jie , Wang Haoqian , He Mingming , Li Nana , Hao Xinyuan , Yang Yajun , Ni Dejiang , Wang Lu , Wang Xinchao
Published: March 2023

Photosystem II repair in chloroplasts is a critical process involved in maintaining a plant’s photosynthetic activity under cold stress. FtsH (filamentation temperature-sensitive H) is an essential metalloprotease that is required for chloroplast photosystem II repair. However, the role of FtsH in tea plants and its regulatory mechanism under cold stress remains elusive. In this study, we cloned a FtsH homolog gene in tea plants, named CsFtsH5, and found that CsFtsH5 was located in the chloroplast and cytomembrane. RT-qPCR showed that the expression of CsFtsH5 was increased with leaf maturity and was significantly induced by light and cold stress. Transient knockdown CsFtsH5 expression in tea leaves using antisense oligonucleotides resulted in hypersensitivity to cold stress, along with higher relative electrolyte leakage and lower Fv/Fm values. To investigate the molecular mechanism underlying CsFtsH5 involvement in the cold stress, we focused on the calcineurin B-like-interacting protein kinase 11 (CsCIPK11), which had a tissue expression pattern similar to that of CsFtsH5 and was also upregulated by light and cold stress. Yeast two-hybrid and dual luciferase (Luc) complementation assays revealed that CsFtsH5 interacted with CsCIPK11. Furthermore, the Dual-Luc assay showed that CsCIPK11-CsFtsH5 interaction might enhance CsFtsH5 stability. Altogether, our study demonstrates that CsFtsH5 is associated with CsCIPK11 and plays a positive role in maintaining the photosynthetic activity of tea plants in response to low temperatures.
Airway and Lung Organoids from Human-Induced Pluripotent Stem Cells Can Be Used to Assess CFTR Conductance
Authors: Demchenko Anna , Kondrateva Ekaterina , Tabakov Vyacheslav , Efremova Anna , Salikhova Diana , Bukharova Tatiana , Goldshtein Dmitry , Balyasin Maxim , Bulatenko Natalia , Amelina Elena , Lavrov Alexander , Smirnikhina Svetlana
Published: March 2023

Airway and lung organoids derived from human-induced pluripotent stem cells (hiPSCs) are current models for personalized drug screening, cell–cell interaction studies, and lung disease research. We analyzed the existing differentiation protocols and identified the optimal conditions for obtaining organoids. In this article, we describe a step-by-step protocol for differentiating hiPSCs into airway and lung organoids. We obtained airway and lung organoids from a healthy donor and from five donors with cystic fibrosis. Analysis of the cellular composition of airway and lung organoids showed that airway organoids contain proximal lung epithelial cells, while lung organoids contain both proximal and distal lung epithelial cells. Forskolin-induced swelling of organoids derived from a healthy donor showed that lung organoids, as well as airway organoids, contain functional epithelial cells and swell after 24 h exposure to forskolin, which makes it a suitable model for analyzing the cystic fibrosis transmembrane conductance regulator (CFTR) channel conductance in vitro. Thus, our results demonstrate the feasibility of generating and characterizing airway and lung organoids from hiPSCs, which can be used for a variety of future applications.
The Role of Inositols in the Hyperandrogenic Phenotypes of PCOS: A Re-Reading of Larner’s Results
Authors: Fedeli Valeria , Catizone Angela , Querqui Alessandro , Unfer Vittorio , Bizzarri Mariano
Published: March 2023

Polycystic ovarian syndrome (PCOS) is the most common endocrinological disorder in women, in which, besides chronic anovulation/oligomenorrhea and ovarian cysts, hyperandrogenism plays a critical role in a large fraction of subjects. Inositol isomers—myo-Inositol and D-Chiro-Inositol—have recently been pharmacologically effective in managing many PCOS symptoms while rescuing ovarian fertility. However, some disappointing clinical results prompted the reconsideration of their specific biological functions. Surprisingly, D-Chiro-Ins stimulates androgen synthesis and decreases the ovarian estrogen pathway; on the contrary, myo-Ins activates FSH response and aromatase activity, finally mitigating ovarian hyperandrogenism. However, when the two isomers are given in association—according to the physiological ratio of 40:1—patients could benefit from myo-Ins enhanced FSH and estrogen responsiveness, while taking advantage of the insulin-sensitizing effects displayed mostly by D-Chiro-Ins. We need not postulate insulin resistance to explain PCOS pathogenesis, given that insulin hypersensitivity is likely a shared feature of PCOS ovaries. Indeed, even in the presence of physiological insulin stimulation, the PCOS ovary synthesizes D-Chiro-Ins four times more than that measured in control theca cells. The increased D-Chiro-Ins within the ovary is detrimental in preserving steroidogenic control, and this failure can easily explain why treatment strategies based upon high D-Chiro-Ins have been recognized as poorly effective. Within this perspective, two factors emerge as major determinants in PCOS: hyperandrogenism and reduced aromatase expression. Therefore, PCOS could no longer be considered a disease only due to increased androgen synthesis without considering the contemporary downregulation of aromatase and FSH receptors. Furthermore, these findings suggest that inositols can be specifically effective only for those PCOS phenotypes featured by hyperandrogenism.
Cheminformatics-Based Study Identifies Potential Ebola VP40 Inhibitors
Authors: Broni Emmanuel , Ashley Carolyn , Adams Joseph , Manu Hammond , Aikins Ebenezer , Okom Mary , Miller Whelton A. , Wilson Michael D. , Kwofie Samuel K.
Published: March 2023

The Ebola virus (EBOV) is still highly infectious and causes severe hemorrhagic fevers in primates. However, there are no regulatorily approved drugs against the Ebola virus disease (EVD). The highly virulent and lethal nature of EVD highlights the need to develop therapeutic agents. Viral protein 40 kDa (VP40), the most abundantly expressed protein during infection, coordinates the assembly, budding, and release of viral particles into the host cell. It also regulates viral transcription and RNA replication. This study sought to identify small molecules that could potentially inhibit the VP40 protein by targeting the N-terminal domain using an in silico approach. The statistical quality of AutoDock Vina’s capacity to discriminate between inhibitors and decoys was determined, and an area under the curve of the receiver operating characteristic (AUC-ROC) curve of 0.791 was obtained. A total of 29,519 natural-product-derived compounds from Chinese and African sources as well as 2738 approved drugs were successfully screened against VP40. Using a threshold of −8 kcal/mol, a total of 7, 11, 163, and 30 compounds from the AfroDb, Northern African Natural Products Database (NANPDB), traditional Chinese medicine (TCM), and approved drugs libraries, respectively, were obtained after molecular docking. A biological activity prediction of the lead compounds suggested their potential antiviral properties. In addition, random-forest- and support-vector-machine-based algorithms predicted the compounds to be anti-Ebola with IC50 values in the micromolar range (less than 25 μM). A total of 42 natural-product-derived compounds were identified as potential EBOV inhibitors with desirable ADMET profiles, comprising 1, 2, and 39 compounds from NANPDB (2-hydroxyseneganolide), AfroDb (ZINC000034518176 and ZINC000095485942), and TCM, respectively. A total of 23 approved drugs, including doramectin, glecaprevir, velpatasvir, ledipasvir, avermectin B1, nafarelin acetate, danoprevir, eltrombopag, lanatoside C, and glycyrrhizin, among others, were also predicted to have potential anti-EBOV activity and can be further explored so that they may be repurposed for EVD treatment. Molecular dynamics simulations coupled with molecular mechanics Poisson–Boltzmann surface area calculations corroborated the stability and good binding affinities of the complexes (−46.97 to −118.9 kJ/mol). The potential lead compounds may have the potential to be developed as anti-EBOV drugs after experimental testing.
Evaluation of the Ion Channel Assembly in a Eukaryotic Cell-Free System Focusing on Two-Pore Domain Potassium Channels K2P
Authors: Ullrich Jessica , Ohlhoff Carsten , Dondapati Srujan Kumar , Zemella Anne , Kubick Stefan
Published: March 2023

Oligomeric ion channels are abundant in nature. However, the recombinant expression in cell culture-based systems remains tedious and challenging due to negative side effects, limiting the understanding of their role in health and disease. Accordingly, in this work, we demonstrate the cell-free synthesis (CFS) as an alternative platform to study the assembly of two-pore domain potassium channels (K2P) within endogenous endoplasmic reticulum-derived microsomes. Exploiting the open nature of CFS, we investigate the cotranslational translocation of TREK-2 into the microsomes and suggest a cotranslational assembly with typical single-channel behavior in planar lipid-bilayer electrophysiology. The heteromeric assembly of K2P channels is a contentious matter, accordingly we prove the successful assembly of TREK-2 with TWIK-1 using a biomolecular fluorescence complementation assay, Western blot analysis and autoradiography. The results demonstrate that TREK-2 homodimer assembly is the initial step, followed by heterodimer formation with the nascent TWIK-1, providing evidence of the intergroup heterodimerization of TREK-2 and TWIK-1 in eukaryotic CFS. Since K2P channels are involved in various pathophysiological conditions, including pain and nociception, CFS paves the way for in-depth functional studies and related pharmacological interventions. This study highlights the versatility of the eukaryotic CFS platform for investigating ion channel assembly in a native-like environment.
CAR T Cell Therapy: A Versatile Living Drug
Authors: De Marco Rodrigo C. , Monzo Hector J. , Ojala Päivi M.
Published: March 2023

After seeing a dramatic increase in the development and use of immunotherapy and precision medicine over the past few decades, oncological care now embraces the start of the adoptive cell therapy (ACT) era. This impulse towards a new treatment paradigm has been led by chimeric antigen receptor (CAR) T cells, the only type of ACT medicinal product to be commercialized so far. Brought about by an ever-growing understanding of cellular engineering, CAR T cells are T lymphocytes genetically modified with an appropriate DNA construct, which endows them with expression of a CAR, a fusion protein between a ligand-specific recognition domain, often an antibody-like structure, and the activating signaling domain of the T cell receptor. Through this genetic enhancement, CAR T cells are engineered from a cancer patient’s own lymphocytes to better target and kill their cancer cells, and the current amassed data on clinical outcomes point to a stream of bright developments in the near future. Herein, from concept design and present-day manufacturing techniques to pressing hurdles and bright discoveries around the corner, we review and thoroughly describe the state of the art in CAR T cell therapy.
Stochastic Dynamic Mass Spectrometric Quantitative and Structural Analyses of Pharmaceutics and Biocides in Biota and Sewage Sludge
Authors: Ivanova Bojidarka
Published: March 2023

Mass spectrometric innovations in analytical instrumentation tend to be accompanied by the development of a data-processing methodology, expecting to gain molecular-level insights into real-life objects. Qualitative and semi-quantitative methods have been replaced routinely by precise, accurate, selective, and sensitive quantitative ones. Currently, mass spectrometric 3D molecular structural methods are attractive. As an attempt to establish a reliable link between quantitative and 3D structural analyses, there has been developed an innovative formula [DSD″,tot=∑inDSD″,i=∑in2.6388.10−17×Ii2¯−Ii¯2] capable of the exact determination of the analyte amount and its 3D structure. It processed, herein, ultra-high resolution mass spectrometric variables of paracetamol, atenolol, propranolol, and benzalkonium chlorides in biota, using mussel tissue and sewage sludge. Quantum chemistry and chemometrics were also used. Results: Data on mixtures of antibiotics and surfactants in biota and the linear dynamic range of concentrations 2–80 ng.(mL)−1 and collision energy CE = 5–60 V are provided. Quantitative analysis of surfactants in biota via calibration equation ln[D″SD] = f(conc.) yields the exact parameter |r| = 0.99991, examining the peaks of BAC-C12 at m/z 212.209 ± 0.1 and 211.75 ± 0.15 for tautomers of fragmentation ions. Exact parameter |r| = 1 has been obtained, correlating the theory and experiments in determining the 3D molecular structures of ions of paracetamol at m/z 152, 158, 174, 301, and 325 in biota.
The Correlation of Apolipoprotein B with Alterations in Specific Fat Depots Content in Adults
Authors: Chen Junye , Li Kang , Shao Jiang , Lai Zhichao , Feng Yuyao , Liu Bao
Published: March 2023

Body mass index (BMI) and blood biomarkers are not enough to predict cardiovascular disease risk. Apolipoprotein B was identified to be associated with cardiovascular disease (CVD) progression. The Dual-energy X-ray Absorption (DXA) results could be considered as a predictor for cardiovascular disease in a more refined way based on fat distribution. The prediction of CVD risk by simple indicators still cannot meet clinical needs. The association of ApoB with specific fat depot features remains to be explored to better co-predict cardiovascular disease risk. An amount of 5997 adults from National Health and Nutrition Examination Survey (NHANES) were enrolled. Their demographic information, baseline clinical condition, blood examination, and DXA physical examination data were collected. Multivariate regression was used to assess the correlation between ApoB and site-specific fat characteristics through different adjusted models. Smooth curve fittings and threshold analysis were used to discover the turning points with 95% confidence intervals. ApoB is positively correlated with arms percent fat, legs percent fat, trunk percent fat, android percent fat, gynoid percent fat, arm circumference and waist circumference after adjustment with covariates for age, gender, race, hypertension, diabetes, hyperlipidemia, coronary heart disease, smoking status and vigorous work activity. The smooth curve fitting and threshold analysis also showed that depot-specific fat had lower turning points of ApoB in both males and females within the normal reference range of ApoB. Meanwhile, females have a lower increase in ApoB per 1% total percent fat and android percent fat than males before the turning points, while females have a higher growth of ApoB per 1% gynoid percent fat than males. The combined specific fat-depot DXA and ApoB analysis could indicate the risk of CVD in advance of lipid biomarkers or DXA alone.
How Single Amino Acid Substitutions Can Disrupt a Protein Hetero-Dimer Interface: Computational and Experimental Studies of the LigAB Dioxygenase from Sphingobium sp. Strain SYK-6
Authors: Rafalowski Angelika , Hassan Bakar A. , Lou Kate , Nguyen Minh Chau , Taylor Erika A.
Published: March 2023

Protocatechuate 4,5-dioxygenase (LigAB) is a heterodimeric enzyme that catalyzes the dioxygenation of multiple lignin derived aromatic compounds. The active site of LigAB is at the heterodimeric interface, with specificity conferred by the alpha subunit and catalytic residues contributed by the beta subunit. Previous research has indicated that the phenylalanine at the 103 position of the alpha subunit (F103α) controls selectivity for the C5 position of the aromatic substrates, and mutations of this residue can enhance the rate of catalysis for substrates with larger functional groups at this position. While several of the mutations to this position (Valine, V; Threonine, T; Leucine, L; and Histidine, H) were catalytically active, other mutations (Alanine, A; and Serine, S) were found to have reduced dimer interface affinity, leading to challenges in copurifing the catalytically active enzyme complex under high salt conditions. In this study, we aimed to experimentally and computationally interrogate residues at the dimer interface to discern the importance of position 103α for maintaining the integrity of the heterodimer. Molecular dynamic simulations and electrophoretic mobility assays revealed a preference for nonpolar/aromatic amino acids in this position, suggesting that while substitutions to polar amino acids may produce a dioxygenase with a useful substrate utilization profile, those considerations may be off-set by potential destabilization of the catalytically active oligomer. Understanding the dimerization of LigAB provides insight into the multimeric proteins within the largely uncharacterized superfamily and characteristics to consider when engineering proteins that can degrade lignin efficiently. These results shed light on the challenges associated with engineering proteins for broader substrate specificity.
Glyphosate and Aminomethylphosphonic Acid (AMPA) Modulate Glutathione S-Transferase in Non-Tumorigenic Prostate Cells
Authors: Borges Dayanne Silva , Vecchi Lara , Barros Deysse Carla Tolentino , Arruda Vinícius Marques , Ferreira Helen Soares Valença , da Silva Matheus Fernandes , Guerra Joyce Ferreira da Costa , Siqueira Raoni Pais , Araújo Thaise Gonçalves
Published: March 2023

Glyphosate (GLY) was developed in the early 1970s and has become the most used broad-spectrum herbicide in the world so far. Its main metabolite is aminomethylphosphonic acid (AMPA), and the accumulation of GLY and its derivative compounds raises some concerns regarding possible health outcomes. In this study, we aimed to evaluate the effects of GLY and AMPA on prostate cell lines by evaluating cell viability, proliferation, gene and protein expression, and cellular pathways involved in the response to oxidative stress. Our results indicated that GLY and AMPA reduced the cell viability of tumorigenic and non-tumorigenic prostate cell lines only at higher concentrations (10 mM GLY and 20 mM AMPA). In contrast, both compounds increased the clonogenicity of non-tumorigenic PNT2 cells, mainly at concentrations below the IC50 (5 mM GLY and 10 mM AMPA). Moreover, treatment of non-tumorigenic cells with low concentrations of GLY or AMPA for 48 h increased GSTM3 expression at both mRNA and protein levels. In contrast, the treatments decrease the GST activity and induced an increase in oxidative stress, mainly at lower concentrations. Therefore, both compounds can cause cellular damage even at lower concentrations in non-tumorigenic PNT2 cells, mainly affecting cell proliferation and oxidative stress.
WHIRLY1 Acts Upstream of ABA-Related Reprogramming of Drought-Induced Gene Expression in Barley and Affects Stress-Related Histone Modifications
Authors: Manh Minh Bui , Ost Charlotte , Peiter Edgar , Hause Bettina , Krupinska Karin , Humbeck Klaus
Published: March 2023

WHIRLY1, a small plant-specific ssDNA-binding protein, dually located in chloroplasts and the nucleus, is discussed to act as a retrograde signal transmitting a stress signal from the chloroplast to the nucleus and triggering there a stress-related gene expression. In this work, we investigated the function of WHIRLY1 in the drought stress response of barley, employing two overexpression lines (oeW1-2 and oeW1-15). The overexpression of WHIRLY1 delayed the drought-stress-related onset of senescence in primary leaves. Two abscisic acid (ABA)-dependent marker genes of drought stress, HvNCED1 and HvS40, whose expression in the wild type was induced during drought treatment, were not induced in overexpression lines. In addition, a drought-related increase in ABA concentration in the leaves was suppressed in WHIRLY1 overexpression lines. To analyze the impact of the gain-of-function of WHIRLY1 on the drought-related reprogramming of nuclear gene expression, RNAseq was performed comparing the wild type and an overexpression line. Cluster analyses revealed a set of genes highly up-regulated in response to drought in the wild type but not in the WHIRLY1 overexpression lines. Among these genes were many stress- and abscisic acid (ABA)-related ones. Another cluster comprised genes up-regulated in the oeW1 lines compared to the wild type. These were related to primary metabolism, chloroplast function and growth. Our results indicate that WHIRLY1 acts as a hub, balancing trade-off between stress-related and developmental pathways. To test whether the gain-of-function of WHIRLY1 affects the epigenetic control of stress-related gene expression, we analyzed drought-related histone modifications in different regions of the promoter and at the transcriptional start sites of HvNCED1 and HvS40. Interestingly, the level of euchromatic marks (H3K4me3 and H3K9ac) was clearly decreased in both genes in a WHIRLY1 overexpression line. Our results indicate that WHIRLY1, which is discussed to act as a retrograde signal, affects the ABA-related reprogramming of nuclear gene expression during drought via differential histone modifications.