bluuebunny/crossref_metadata_2025 huggingface dataset:
The Effects of Ischemia During Rest Intervals on Bar Velocity in the Bench Press Exercise With Different External Loads
Authors: Jarosz Jakub , Trybulski Robert , Krzysztofik Michał , Tsoukos Athanasios , Filip-Stachnik Aleksandra , Zajac Adam , Bogdanis Gregory C. , Wilk Michal
Published: August 2021

The main aim of the present study was to evaluate the acute effects of ischemia used during rest periods on bar velocity changes during the bench press exercise at progressive loads, from 20 to 90% of 1RM. Ten healthy resistance trained men volunteered for the study (age = 26.3 ± 4.7 years; body mass = 89.8 ± 6.3 kg; bench press 1RM = 142.5 ± 16.9 kg; training experience = 7.8 ± 2.7 years). During the experimental sessions the subjects performed the bench press exercise under two different conditions, in a randomized and counterbalanced order: (a) ischemia condition, with ischemia applied before the first set and during every rest periods between sets, and (b) control condition where no ischemia was applied. During each experimental session eight sets of the bench press exercise were performed, against loads starting from 20 to 90% 1RM, increased progressively by 10% in each subsequent set. A 3-min rest interval between sets was used. For ischemia condition the cuffs was applied 3 min before the first set and during every rest period between sets. Ischemia was released during exercise. The cuff pressure was set to ∼80% of full arterial occlusion pressure. The two-way repeated measures ANOVA showed a statistically significant interaction effect for peak bar velocity (p = 0.04) and for mean bar velocity (p = 0.01). There was also a statistically significant main effect of condition for peak bar velocity (p < 0.01) but not for mean bar velocity (p = 0.25). The post hoc analysis for interaction showed significantly higher peak bar velocity for the ischemia condition compared to control at a load of 20% 1RM (p = 0.007) and at a load of 50% 1RM (p = 0.006). The results of the present study indicate that ischemia used before each set even for a brief duration of <3 min, has positive effects on peak bar velocity at light loads, but it is insufficient to induce such effect on higher loads.
Improvement of the LbCas12a-crRNA System for Efficient Gene Targeting in Tomato
Authors: Vu Tien Van , Doan Duong Thi Hai , Tran Mil Thi , Sung Yeon Woo , Song Young Jong , Kim Jae-Yean
Published: August 2021

Plant gene targeting (GT) can be utilized to precisely replace up to several kilobases of a plant genome. Recent studies using the powerful clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) nucleases significantly improved plant GT efficiency. However, GT for loci without associated selection markers is still inefficient. We previously utilized Lachnospiraceae bacterium Cas12a (LbCas12a) in combination with a replicon for tomato GT and obtained high GT efficiency with some selection markers. In this study, we advance our GT system by inhibiting the cNHEJ pathway with small chemical molecules such as NU7441. Further optimization of the GT is also possible with the treatment of silver nitrate possibly via its pronounced actions in ethylene inhibition and polyamine production. Importantly, the GT efficiency is significantly enhanced with the use of a temperature-tolerant LbCas12a (ttLbCas12a) that is capable of performing target cleavage even at low temperatures. Targeted deep sequencing, as well as conventional methods, are used for the assessment of the editing efficiency at both cell and plant levels. Our work demonstrates the significance of the selection of gene scissors, the appropriate design and number of LbCas12a crRNAs, the use of chemical treatments, and the establishment of favorable experimental conditions for further enhancement of plant HDR to enable efficient GT in tomato.
Actions and the Self: I Give, Therefore I am?
Authors: Regner Tobias , Matthey Astrid
Published: August 2021

Self-signaling models predict less selfish behavior in a probabilistic giving setting as individuals are expected to invest in a pro-social identity. However, there is also substantial evidence that people tend to exploit situational excuses for selfish choices (for instance, uncertainty) and behave more selfishly. We contrast these two motivations (identity management and self-deception) experimentally in order to test which one is more prevalent in a reciprocal giving setting. Trustees' back transfer choices are elicited for five different transfer levels of the trustor. Moreover, we ask trustees to provide their back transfer schedule for different scenarios that vary the implementation probability of the back transfer. This design allows us to identify subjects who reciprocate and analyze how these reciprocators respond when self-image relevant factors are varied. Our results indicate that self-deception is prevalent when subjects make the back transfer choice. Twice as many subjects seem to exploit situational excuses than subjects who appear to invest in a pro-social identity.

JEL classifications: C72, C91, D80, D91
Public Opinion and Expectations: Development of Public Health Education in China After COVID-19 Pandemic
Authors: Shen Xin , Li Crystal Jingru , Dong Tianyi , Cao Hui , Feng Jing , Lei Zihui , Wang Zijian , Han Xiaotong , Lv Chuanzhu , Gan Yong
Published: August 2021

Background: Policymakers must promote the development of public health education and human resources. As a feature of the political environment, public opinion is essential for policy-making, but virtually the attitudes of Chinese citizens toward human resources development in public health is unknown.

Methods: This study conducted a crosssectional survey from February 4, 2021 to February 26, 2021 in China. We adopted a convenient sampling strategy to recruit participators. Participants filled out the questions, which assess the attitudes of the expanding public health professionals. A logistic regression analysis was given to identify the predictors associated with the attitudes of the subjects.

Results: There were 2,361 residents who have finished our questionnaire. Chinese residents who lived in urban (OR = 1.293, 95% CI = 1.051–1.591), “themselves or relatives and friends have participated in relevant epidemic prevention work” (OR = 1.553, 95% CI = 1.160–2.079), “themselves or family members engaged in medical-related work” (OR = 1.468, 95% CI = 1.048–2.056), and those who “were aware of public health before the outbreak of COVID-19” (OR = 1.428, 95% CI = 1.125–1.812) were more likely to support the promotion of public health education and training.

Conclusions: The present study found that 74.50% of Chinese citizens supported the promotion of public health education and training in China, in which economic status, personal perception, and comprehension are the crucial factors that influence public opinion. COVID-19 has aroused the attention of Chinese residents to public health education, with only 22.11% of residents being aware of public health before the outbreak of COVID-19. The COVID-19 pandemic has profound implications for human society. Literally, this impact will feed back into future public health policies based on public opinion. This innovative perspective will also help us better understand the potential social impact of COVID-19 on human resources and development for health in the modern world.
Universities as Partners in Primary Health Care Innovation
Authors: Majewski Henryk
Published: August 2021

Universities have a unique role in the health ecosystem as providers of trained staff and discoverers of health innovations. However, often they sit in silos waiting for their rare blockbuster discoveries to change clinical care or seeing health services simply as future employers of their graduates or clinical trial sites. It is a transactional and targetted relationship. This present case study is of a primary health service Access Health and Community (AccessHC) in Australia and its university partner Swinburne University of Technology. Together they established a Kickstart Program which was to provide seed funding for small joint innovation projects generated by both organisations. One project exemplifies the approach. Swinburne who has a Design School was encouraged through the Kickstart Program to design a clinical waiting room of the future. This project started with a needs analysis. The written report was to inform the design. University staff linked with their internal University animations expertise to better communicate the needs analysis. The “Access me Not” animation was created, unknown to the staff at AccessHC. At initial presentation, the way the animation communicated was not imaginable by AccessHC. “Access me not” was submitted for the 2018 International Design Awards and received an honourable mention. However, the AccessHC staff saw other uses for the approach and contacted Swinburne to design a client journey animation for the newly introduced National Disability Scheme (NDIS). The co design produced an animation of immense help to parents in navigating the scheme for complex and chronic disability care and for AccessHC the scripting served as a framework to develop it new internal NDIS care systems and processes. The Swinburne team is now producing health navigation animations for the State Department of Health and Human Services. The Kickstart Program was an engagement strategy that has produced a set of health communication tools that the health service could not have envisaged and which the University could not have imagined an application. Small low risk seed funding can indeed introduce innovations and create beneficial relationships between health services and universities.
Regenerative Almond Production Systems Improve Soil Health, Biodiversity, and Profit
Authors: Fenster Tommy L. D. , Oikawa Patricia Y. , Lundgren Jonathan G.
Published: August 2021

Regenerative agriculture aims to improve soil health and promote biodiversity while producing nutritious food profitably. Almonds are the dominant crop in California agriculture in terms of acreage and revenue generated. We examined the soil health, biodiversity, yield, and profit of regenerative and conventional almond production systems that represented farmer-derived best management practices. Regenerative practices included abandoning some or all synthetic agrichemicals, planting perennial ground covers, integrating livestock, maintaining non-crop habitat, and using composts and compost teas. Total soil carbon (TSC), soil organic matter (SOM), total soil nitrogen (TSN), total soil phosphorous, calcium, sulfur, and soil health test scores were all significantly greater in regenerative soils. Water infiltrated regenerative soils six-fold faster than conventional soils. Total microbial biomass, total bacterial biomass, Gram+ bacteria, and Actinobacteria were significantly greater in regenerative soils. There was more plant biomass, species diversity, and percent cover in regenerative orchards. Invertebrate richness and diversity, and earthworm abundance and biomass were significantly greater in regenerative orchards. Pest populations, yields, and nutrient density of the almonds were similar in the two systems. Profit was twice as high in the regenerative orchards relative to their conventional counterparts. No one practice was responsible for the success of regenerative farms; their success was the result of simultaneously combining multiple regenerative practices into a single, functional farm system. This style of farming may assist in combatting planetary scale problems (e.g., climate change, biodiversity loss, agricultural pollution, chronic human health problems, and declining rural communities) while making farms more profitable and resilient.
Nuclear Magnetic Resonance-Based Metabolomic Analysis Reveals Physiological Stage, Breed, and Diet Effects on the Intramuscular Metabolism of Amino Acids and Related Nutrients in Pigs
Authors: Liu Yingying , He Qinghua , Azad Md. Abul Kalam , Xiao Yi , Yin Yulong , Kong Xiangfeng
Published: August 2021

Skeletal muscle is a complex tissue that exhibits considerable plasticity in response to nutrients, animal, or its growth stage, but the underlying mechanisms are largely unknown. This study was conducted to evaluate the effects of physiological stage, breed, and diet on the metabolome of the skeletal muscle of pigs. Ninety-six barrows, including 48 purebred Bama mini-pigs, representing the fat type, and 48 Landrace pigs, representing the lean type, were randomly assigned to either a low- or adequate-protein diet (n = 24 per group). The experimental period commenced at 5 weeks of age and extended to the finishing period. Psoas major muscles (PMMs) were collected at the nursery, growing, and finishing stages; and the contents of amino acids (AAs), fatty acids (FAs), and metabolites were analyzed using a nuclear magnetic resonance-based approach. Results showed that most AAs and monounsaturated FAs (MUFAs; including C16:1 and C18:1) contents were increased (p < 0.05) gradually, while those of polyunsaturated FAs (including C18:2, C20:4n−6, C20:5n−3, and C22:6n−3) were decreased (p < 0.05) in the PMM with increasing age. Compared with Landrace pigs, Bama mini-pigs had higher (p < 0.05) contents of flavor-related AAs (including methionine, phenylalanine, tyrosine, leucine, and serine) in the nursery and growing stages and higher (p < 0.05) percentages of saturated FAs and MUFAs throughout the trial. Dietary protein levels affected the muscular profiles of AAs and FAs in an age-dependent manner. In addition, the adequate-protein diet increased (p < 0.05) the muscular contents of α-ketoglutarate in the two breeds. These findings indicate that the dynamic profiles of AAs, FAs, and metabolites in pig muscle tissues are regulated by breed, diet, and physiological stage.
Spatial Layout of Cotton Seed Production Based on Hierarchical Classification: A Case Study in Xinjiang, China
Authors: Niu Yingnan , Xie Gaodi , Xiao Yu , Qin Keyu , Liu Jingya , Wang Yangyang , Gan Shuang , Huang Mengdong , Liu Jia , Zhang Caixia , Zhang Changshun
Published: August 2021

Cotton seed production is the main form of agriculture in Xinjiang, China. Unreasonable distribution of cotton seed production results in a waste of water, land, and human resources. In this study, we established a hierarchical classification integrating method; investigated the spatial suitability of climate, land and water resources, and infrastructure; examined the production risk and planting history; and integrated spatial suitability and production risk and history to produce the spatial layout of seed production for early-maturing cotton (EMC), early–medium-maturing cotton (EMMC), and long staple cotton (LSC) in Xinjiang. The results indicated that the appropriate areas for EMC, EMMC, and LSC seed production are 6.4 × 105, 5.5 × 105, and 3.6 × 105 ha, respectively. By combining the suitable areas of seed production for the three cotton species, we concluded that the superior and most suitable area for producing cotton seed of EMC, EMMC, or LSC is located in the western Tarim Basin. The sub-suitable area for cotton seed production of EMC, EMMC, or LSC is mostly distributed in the western and northern Tarim Basin. This research provides a good solution to the selection of cotton seed production base; however, adoption will depend on the actual preference and market factors.
PPO2 Mutations in Amaranthus palmeri: Implications on Cross-Resistance
Authors: Carvalho-Moore Pâmela , Rangani Gulab , Heiser James , Findley Douglas , Bowe Steven J. , Roma-Burgos Nilda
Published: August 2021

In Arkansas, resistance to protoporphyrinogen IX oxidase (PPO)-inhibiting herbicides in Amaranthus palmeri S. Wats. is mainly due to target site mutations. Although A. palmeri PPO-mutations are well investigated, the cross-resistance that each ppo mutant endows to weed populations is not yet well understood. We aimed to evaluate the response of PPO-resistant A. palmeri accessions, harboring the ppo2 mutations ΔG210 and G399A, to multiple PPO-inhibiting herbicides. Six resistant and one susceptible field accessions were subjected to a dose–response assay with fomesafen, and selected survivors from different fomesafen doses were genotyped to characterize the mutation profile. The level of resistance to fomesafen was determined and a cross-resistance assay was conducted with 1 and 2 times the labeled doses of selected PPO herbicides. The accession with higher predicted dose to control 50% of the population (ED50) had a higher frequency of ΔG210-homozygous survivors. Survivors harboring both mutations, and those that were ΔG210-homozygous, incurred less injury at the highest fomesafen rate tested (1120 g ai ha−1). The populations with a high frequency of ΔG210-homozygous survivors, and those with individuals harboring ΔG210 + G399A mutations, exhibited high potential for cross-resistance to other PPO herbicides. The new PPO–herbicide chemistries (saflufenacil, trifludimoxazin) generally controlled the PPO-resistant populations.
Licorice, Doum, and Banana Peel Extracts Inhibit Aspergillus flavus Growth and Suppress Metabolic Pathway of Aflatoxin B1 Production
Authors: Youssef Nesrine H. , Qari Sameer H. , Matar Saleh , Hamad Najwa A. , Dessoky Eldessoky S. , Elshaer Moustafa M. , Sobhy Sherien , Abdelkhalek Ahmed , Zakaria Hossam M. , Heflish Ahmed A. , Elsamra Ibrahim A. , Behiry Said I.
Published: August 2021

Three different concentrations of four (ethanol, acetone, methanol, and diethyl ether) extracts of licorice, doum, and banana peel were evaluated for antifungal and antimycotoxigenic efficiency against a maize aflatoxigenic fungus, Aspergillus flavus. Among them, the licorice diethyl ether 75% extract was intensely active, showing the best wet and dry weight inhibition and exhibiting the highest efficacy ratio (91%). Regarding aflatoxin B1 (AFB1) production, all the plant extracts tested were effective against AFB1 production after one month of maize storage, with average efficacy ratios ranging from 74.1% to 97.5%. At the same time, Thiram fungicide exhibited an efficacy ratio of 20.14%. The relative expression levels of three structural genes (aflD, aflP, and aflQ) and two regulatory genes (aflR and aflS) were significantly downregulated when compared to untreated maize grains or Thiram-treated maize grains. The doum diethyl ether 75% peel extract showed the highest total phenolic content (60.48 mg GAE/g dry extract wt.) and antioxidant activity (84.71 μg/mL). GC–MS analysis revealed that dimethoxycinnamic acid, aspartic acid, valproic acid, and linoleic acid might imbue the extracts with antioxidant capacities in relation to fungal growth and aflatoxin biosynthesis. Finally, the results suggest that the three plant extracts can be considered a promising source for developing potentially effective and environmentally safer alternative ways to control aflatoxin formation, thus creating a potentially protective method for grain storage.
The Antimicrobial Properties of Nanotitania Extract and Its Role in Inhibiting the Growth of Klebsiella pneumonia and Haemophilus influenza
Authors: Harun Ahmad Mukifza , Noor Nor Farid Mohd , Zaid Awatief , Yusoff Mohamad Ezany , Shaari Ramizu , Affandi Nor Dalila Nor , Fadil Fatirah , Rahman Mohd Azizi Abdul , Alam Mohammad Khursheed
Published: August 2021

Titanium dioxide (TiO2) is an antimicrobial agent which is considered of potential value in inhibiting the growth of multiple bacteria. Klebsiella pneumonia and Haemophilus influenza are two of the most common respiratory infection pathogens, and are the most. Klebsiella pneumonia causes fatal meningitis, while Haemophilus influenza causes mortality even in younger patients. Both are associated with bacteremia and mortality. The purpose of this study was to test a new antibacterial material, namely nanotitania extract combined with 0.03% silver that was developed at Universiti Malaysia Sabah (UMS) and tested against K. pneumonia and H. influenza. The nanoparticles were synthesized through a modified hydrothermal process, combined with molten salt and proven to have excellent crystallinity, with the band-gap energy falling in the visible light spectrum. The nanoparticle extract was tested using a macro-dilutional method, which involved combining it with 0.03% silver solution during the process of nanoparticle synthesis and then introducing it to the bacteria. A positive control containing the bacteria minus the nanoparticles extract was also prepared. 25 mg/mL, 12.5 mg/mL, and 6.25 mg/mL concentrations of the samples were produced using the macro dilution method. After adding the bacteria to multiple concentrations of nanoparticle extract, the suspensions were incubated for 24 h at a temperature of 37 °C. The suspensions were then spread on Mueller-Hinton agar (K. pneumonia) and chocolate blood agar (H. influenza), where the growth of bacteria was observed after 24 h. Nanoparticle extract in combination with silver at 0.03% was proven to have potential as an antimicrobial agent as it was able to inhibit H. influenza at all concentrations. Furthermore, it was also shown to be capable of inhibiting K. pneumonia at concentrations of 25 mg/mL and 50 mg/mL. In conclusion, the nanoparticle extract, when tested using a macro-dilutional method, displayed antimicrobial properties which were proven effective against the growth of both K. pneumonia and H. influenza.
Effectiveness of Combined Stretching and Strengthening Exercise Using Rehabilitation Exercise System with a Linear Actuator and MR Damper on Static and Dynamic Sitting Postural Balance: A Feasibility Study
Authors: Jung Ji-Yong , Yang Chang-Min , Kim Jung-Ja
Published: August 2021

Postural imbalance induced by prolonged sitting can be improved by exercise therapy. The aim of study was to evaluate the influence of combined stretching and strengthening exercise using rehabilitation exercise system with a linear actuator and MR damper on static and dynamic sitting postural balance. Twelve subjects who sit almost 10 h a day participated in this study. The rehabilitation exercise system with a linear actuator and MR damper was manufactured to provide stretching and strengthening exercise. All subjects were asked to perform an exercise program that was designed to enhance postural balance by stretching the tight muscle and strengthening the weakened muscle. Body pressure distributions were analyzed for mean force and mean pressure using a seat sensor system. Trunk muscle activities were measured by attaching surface electrodes to the thoracic erector spinae, lumbar erector spinae, and lumbar multifidus muscle. All data were divided into two regions (dominant and non-dominant side) under four conditions: no pelvic tilt, lateral pelvic tilt, anterior pelvic tilt, and posterior pelvic tilt. Body pressure distributions and trunk muscle activities were compared between before and after performing exercise under static and dynamic sitting conditions. Both in static and dynamic sitting conditions, there were significant differences in body pressure distributions and trunk muscle activities between the DS and NDS before performing the exercise (p < 0.01). After performing exercise, the body pressure distributions increased on the dominant side while those decreased on the non-dominant side significantly (p < 0.01). In addition, the activities of all trunk muscles on the non-predominant side increased significantly (p < 0.01 and p < 0.05). These results showed that postural balance was improved by decreasing the differences in body pressure distribution and trunk muscle activity between the dominant and non-dominant side after performing exercise. From the results of this study, we concluded that the rehabilitation exercise system with a linear actuator and MR damper is suitable for providing combined stretching and strengthening exercise, and it could be helpful to maintain correct posture by enhancing postural balance during sitting.
Debris Removal by Activation of Endodontic Irrigants in Complex Root Canal Systems: A Standardized In-Vitro-Study
Authors: Widbiller Matthias , Keim Lukas , Schlichting Ralf , Striegl Birgit , Hiller Karl-Anton , Jungbauer Rebecca , Buchalla Wolfgang , Galler Kerstin M.
Published: August 2021

Aim of the study was to develop a standardized model system to investigate endodontic irrigation techniques and assess the efficiency of different activation methods on the removal of hard tissue debris in complex root canal systems. Mesial roots of mandibular molars were firstly scanned by micro-computed tomography (µCT) and allocated to three groups of irrigant activation: sonic activation (EDDY, VDW, Munich, Germany), laser activation (AutoSWEEPS, FOTONA, Ljubljana, Slovenia) and conventional needle irrigation (control). Roots were fixed in individual 3D-printed holders to facilitate root canal enlargement under constant irrigation with NaOCl (5%). To enable standardized quantification of remaining debris, BaSO4-enriched dentine powder was compacted into the canals, followed by another µCT-scan. The final irrigation was performed using 17% ethylenediaminetetraacetic acid (EDTA) and 5% sodium hypochlorite (NaOCl) with the respective activation method, and the volume of remaining artificial debris was quantified after a final µCT-scan. The newly developed model system allowed for reliable, reproducible and standardized assessment of irrigation methods. Activation of the irrigant proved to be significantly more effective than conventional needle irrigation regarding the removal of debris, which persisted particularly in the apical third of the root canal in the control group. The efficiency of irrigation was significantly enhanced with laser- and sonic-based activation, especially in the apical third.
Development Workflow for Manifolds and Fluid Components Based on Laser Powder Bed Fusion
Authors: Rolinck Nicolas , Schmitt Matthias , Schneck Matthias , Schlick Georg , Schilp Johannes
Published: August 2021

Through its unique characteristics, additive manufacturing yields great potential for designing fluid components with increased performance characteristics. These potentials in advanced design, functional structure, and manufacturing are not easily realized. Therefore, the present study proposes a holistic development methodology for fluid components with a specific focus on hydraulic manifolds. The methodology aims to lead the designer from the specification of the task, through a step-by-step embodied design, to a technical and economic evaluation of the optimized, first-time manufactured part. A case study applies the proposed methodology to a part of a rail-vehicle braking application. Through its application, a significant reduction in weight, size, as well as significant contributions to the company’s AM strategy can be assigned to the part. At the same time, increased direct manufacturing costs are identified. Based on the increased performance characteristics of the resulting design and the holistic foundation of the subsequent economic decisions, a satisfying efficiency can be allocated to the proposed methodology.
Modelling the Effect of Temperature on the Initial Decline during the Lag Phase of Geotrichum candidum
Authors: Valík Ľubomír , Šipošová Petra , Koňuchová Martina , Medveďová Alžbeta
Published: August 2021

The study of lag phase provides essential knowledge for food quality control. With respect to significance of Geotrichum candidum in the food context, the aim of this study was to quantitatively characterize the relationship between temperature (6–25 °C) and initial decline period during G. candidum lag phase. The decrease in G. candidum cells in the lag phase was primary modelled by Weibull’s model to define the first-decimal reduction time (δ). Subsequently, the lag death rate (LDR) values were recalculated from δ and further modelled by using Arrhenius equations, as well as a square root model, and the models’ suitability was proven by selected statistical indices. The square root model with the estimated parameters b = 0.016 °C−1 h−0.5 and Tmin = −0.72 °C showed better indices relating to goodness of fit based on a low root mean sum of square error (RMSE = 0.028 log CFU mL−1), a higher coefficient of determination (R2 = 0.978), and the lowest value of AIC (AIC = −38.65). The present study provides a solution to the possible application of secondary predictive models to the death rate dependence on temperature during the microbial lag phase. Despite limited practical importance, under specific conditions, it is possible to consider its use, for example, in exposure assessment.
Advances in Machine Fault Diagnosis
Authors: Vaimann Toomas
Published: August 2021

The growing need for intelligent machines, the outreach for more efficient use of the machines in industry, and the development of Industry 4 [...]
Extreme Weather Events Affect Agronomic Practices and Their Environmental Impact in Maize Cultivation
Authors: Marković Monika , Šoštarić Jasna , Josipović Marko , Atilgan Atilgan
Published: August 2021

Sustainable and profitable crop production has become a challenge due to frequent weather extremes, where unstable crop yields are often followed by the negative impacts of agronomic practices on the environment, i.e., nitrate leaching in irrigated and nitrogen (N)-fertilized crop production. To study this issue, a three-year field study was conducted during quite different growing seasons in terms of weather conditions, i.e., extremely wet, extremely dry, and average years. Over three consecutive years, the irrigation and N fertilizers rates were tested for their effect on grain yield and composition, i.e., protein, starch, and oil content of the maize hybrids; soil N level (%); and nitrate leaching. The results showed that the impact of the tested factors and their significance was year- or weather-condition-dependent. The grain yield result stood out during the extremely wet year, where the irrigation rate reduced the grain yield by 7.6% due to the stress caused by the excessive amount of water. In the remainder of the study, the irrigation rate expectedly increased the grain yield by 13.9% (a2) and 20.8% (a3) in the extremely dry year and 22.7% (a2) and 39.5% (a3) during the average year. Regardless of the weather conditions, the N fertilizer rate increased the grain yield and protein content. The soil N level showed a typical pattern, where the maximum levels were at the beginning of the study period and were higher as the N fertilizer rate was increased. Significant variations in the soil N level were found between weather conditions (r = −0.719) and N fertilizer rate (r = 0.401). Nitrate leaching losses were expectedly found for irrigation and N fertilizer treatments with the highest rates (a3b3 = 79.8 mg NO3− L).
Privacy Preserving Classification of EEG Data Using Machine Learning and Homomorphic Encryption
Authors: Popescu Andreea Bianca , Taca Ioana Antonia , Nita Cosmin Ioan , Vizitiu Anamaria , Demeter Robert , Suciu Constantin , Itu Lucian Mihai
Published: August 2021

Data privacy is a major concern when accessing and processing sensitive medical data. A promising approach among privacy-preserving techniques is homomorphic encryption (HE), which allows for computations to be performed on encrypted data. Currently, HE still faces practical limitations related to high computational complexity, noise accumulation, and sole applicability the at bit or small integer values level. We propose herein an encoding method that enables typical HE schemes to operate on real-valued numbers of arbitrary precision and size. The approach is evaluated on two real-world scenarios relying on EEG signals: seizure detection and prediction of predisposition to alcoholism. A supervised machine learning-based approach is formulated, and training is performed using a direct (non-iterative) fitting method that requires a fixed and deterministic number of steps. Experiments on synthetic data of varying size and complexity are performed to determine the impact on runtime and error accumulation. The computational time for training the models increases but remains manageable, while the inference time remains in the order of milliseconds. The prediction performance of the models operating on encoded and encrypted data is comparable to that of standard models operating on plaintext data.
Mitochondrial Uncoupling Proteins (UCP1-UCP3) and Adenine Nucleotide Translocase (ANT1) Enhance the Protonophoric Action of 2,4-Dinitrophenol in Mitochondria and Planar Bilayer Membranes
Authors: Žuna Kristina , Jovanović Olga , Khailova Ljudmila , Škulj Sanja , Brkljača Zlatko , Kreiter Jürgen , Kotova Elena , Vazdar Mario , Antonenko Yuri , Pohl Elena
Published: August 2021

2,4-Dinitrophenol (DNP) is a classic uncoupler of oxidative phosphorylation in mitochondria which is still used in “diet pills”, despite its high toxicity and lack of antidotes. DNP increases the proton current through pure lipid membranes, similar to other chemical uncouplers. However, the molecular mechanism of its action in the mitochondria is far from being understood. The sensitivity of DNP’s uncoupling action in mitochondria to carboxyatractyloside, a specific inhibitor of adenine nucleotide translocase (ANT), suggests the involvement of ANT and probably other mitochondrial proton-transporting proteins in the DNP’s protonophoric activity. To test this hypothesis, we investigated the contribution of recombinant ANT1 and the uncoupling proteins UCP1-UCP3 to DNP-mediated proton leakage using the well-defined model of planar bilayer lipid membranes. All four proteins significantly enhanced the protonophoric effect of DNP. Notably, only long-chain free fatty acids were previously shown to be co-factors of UCPs and ANT1. Using site-directed mutagenesis and molecular dynamics simulations, we showed that arginine 79 of ANT1 is crucial for the DNP-mediated increase of membrane conductance, implying that this amino acid participates in DNP binding to ANT1.
Lytic Polysaccharide Monooxygenases as Chitin-Specific Virulence Factors in Crayfish Plague
Authors: Sabbadin Federico , Henrissat Bernard , Bruce Neil C. , McQueen-Mason Simon J.
Published: August 2021

The oomycete pathogen Aphanomyces astaci, also known as “crayfish plague”, is an obligate fungal-like parasite of freshwater crustaceans and is considered responsible for the ongoing decline of native European crayfish populations. A. astaci is thought to secrete a wide array of effectors and enzymes that facilitate infection, however their molecular mechanisms have been poorly characterized. Here, we report the identification of AA15 lytic polysaccharide monooxygenases (LPMOs) as a new group of secreted virulence factors in A. astaci. We show that this enzyme family has greatly expanded in A. astaci compared to all other oomycetes, and that it may facilitate infection through oxidative degradation of crystalline chitin, the most abundant polysaccharide found in the crustacean exoskeleton. These findings reveal new roles for LPMOs in animal–pathogen interactions, and could help inform future strategies for the protection of farmed and endangered species.
A Review of Multiple Mitochondrial Dysfunction Syndromes, Syndromes Associated with Defective Fe-S Protein Maturation
Authors: Lebigot Elise , Schiff Manuel , Golinelli-Cohen Marie-Pierre
Published: August 2021

Mitochondrial proteins carrying iron-sulfur (Fe-S) clusters are involved in essential cellular pathways such as oxidative phosphorylation, lipoic acid synthesis, and iron metabolism. NFU1, BOLA3, IBA57, ISCA2, and ISCA1 are involved in the last steps of the maturation of mitochondrial [4Fe-4S]-containing proteins. Since 2011, mutations in their genes leading to five multiple mitochondrial dysfunction syndromes (MMDS types 1 to 5) were reported. The aim of this systematic review is to describe all reported MMDS-patients. Their clinical, biological, and radiological data and associated genotype will be compared to each other. Despite certain specific clinical elements such as pulmonary hypertension or dilated cardiomyopathy in MMDS type 1 or 2, respectively, nearly all of the patients with MMDS presented with severe and early onset leukoencephalopathy. Diagnosis could be suggested by high lactate, pyruvate, and glycine levels in body fluids. Genetic analysis including large gene panels (Next Generation Sequencing) or whole exome sequencing is needed to confirm diagnosis.
Metformin Is a Pyridoxal-5′-phosphate (PLP)-Competitive Inhibitor of SHMT2
Authors: Tramonti Angela , Cuyàs Elisabet , Encinar José , Pietzke Matthias , Paone Alessio , Verdura Sara , Arbusà Aina , Martin-Castillo Begoña , Giardina Giorgio , Joven Jorge , Vazquez Alexei , Contestabile Roberto , Cutruzzolà Francesca , Menendez Javier
Published: August 2021

The anticancer actions of the biguanide metformin involve the functioning of the serine/glycine one-carbon metabolic network. We report that metformin directly and specifically targets the enzymatic activity of mitochondrial serine hydroxymethyltransferase (SHMT2). In vitro competitive binding assays with human recombinant SHMT1 and SHMT2 isoforms revealed that metformin preferentially inhibits SHMT2 activity by a non-catalytic mechanism. Computational docking coupled with molecular dynamics simulation predicted that metformin could occupy the cofactor pyridoxal-5′-phosphate (PLP) cavity and destabilize the formation of catalytically active SHMT2 oligomers. Differential scanning fluorimetry-based biophysical screening confirmed that metformin diminishes the capacity of PLP to promote the conversion of SHMT2 from an inactive, open state to a highly ordered, catalytically competent closed state. CRISPR/Cas9-based disruption of SHMT2, but not of SHMT1, prevented metformin from inhibiting total SHMT activity in cancer cell lines. Isotope tracing studies in SHMT1 knock-out cells confirmed that metformin decreased the SHMT2-channeled serine-to-formate flux and restricted the formate utilization in thymidylate synthesis upon overexpression of the metformin-unresponsive yeast equivalent of mitochondrial complex I (mCI). While maintaining its capacity to inhibit mitochondrial oxidative phosphorylation, metformin lost its cytotoxic and antiproliferative activity in SHMT2-null cancer cells unable to produce energy-rich NADH or FADH2 molecules from tricarboxylic acid cycle (TCA) metabolites. As currently available SHMT2 inhibitors have not yet reached the clinic, our current data establishing the structural and mechanistic bases of metformin as a small-molecule, PLP-competitive inhibitor of the SHMT2 activating oligomerization should benefit future discovery of biguanide skeleton-based novel SHMT2 inhibitors in cancer prevention and treatment.
Prevalence of Germline Variants in a Large Cohort of Japanese Patients with Pheochromocytoma and/or Paraganglioma
Authors: Yonamine Masato , Wasano Koichiro , Aita Yuichi , Sugasawa Takehito , Takahashi Katsutoshi , Kawakami Yasushi , Shimano Hitoshi , Nishiyama Hiroyuki , Hara Hisato , Naruse Mitsuhide , Okamoto Takahiro , Matsuda Tadashi , Kosugi Shinji , Horiguchi Kazuhiko , Tanabe Akiyo , Watanabe Atsushi , Kimura Noriko , Nakamura Eijiro , Sakurai Akihiro , Shiga Kiyoto , Takekoshi Kazuhiro
Published: August 2021

The high incidence of germline variants in pheochromocytoma and paraganglioma (PPGL) has been reported mainly in Europe, but not among Japanese populations in Asia. We aimed to study the prevalence of germline variants in Japanese PPGL patients and the genotype–phenotype correlation. We examined 370 PPGL probands, including 43 patients with family history and/or syndromic presentation and 327 patients with apparently sporadic (AS) presentation. Clinical data and blood samples were collected, and the seven major susceptibility genes (MAX, SDHB, SDHC, SDHD, TMEM127, VHL, and RET) were tested using Sanger sequencing. Overall, 120/370 (32.4%) patients had pathogenic or likely pathogenic variants, with 81/327 (24.8%) in AS presentation. SDHB was the most frequently mutated gene (57, 15.4%), followed by SDHD (27, 7.3%), and VHL (18, 4.9%). The incidence of metastatic PPGL was high in SDHB carriers (21/57, 36.8%). A few unique recurrent variants (SDHB c.137G>A and SDHB c.470delT) were detected in this Japanese cohort, highlighting ethnic differences. In summary, almost a quarter of patients with apparently sporadic PPGL in Japan harboured germline variants of the targeted genes. This study reinforces the recommendation in Western guidelines to perform genetic testing for PPGL and genotype-based clinical decision-making in the Japanese population.
Salvage Autologous Stem Cell Transplantation in Daratumumab-Refractory Multiple Myeloma
Authors: Yarlagadda Lakshmi , Gundarlapalli Sravani , Parikh Richa , Landes Reid D. , Kottarathara Mathew , Ogunsesan Yetunde , Hoque Shadiqul , Mitma Angel A. , Bailey Clyde , Hill Kerri M. , Thanendrarajan Sharmilan , Graziutti Monica , Mohan Meera , Zangari Maurizio , van Rhee Frits , Tricot Guido , Schinke Carolina
Published: August 2021

Daratumumab, a CD38-targeting monoclonal antibody, has significantly improved survival rates in multiple myeloma (MM), yet patients who progress on Daratumumab have dismal clinical outcomes with an overall median of less than 10 months. While emerging novel modalities have shown promising results, the current study explores the use of high-dose chemotherapy followed by autologous stem cell transplantation (ASCT) in heavily pretreated Daratumumab-refractory MM patients. We retrospectively investigated the outcome of 69 consecutive patients who received upfront ASCT. The median progression-free survival (PFS) for the entire patient cohort was 7.2 months with a median overall survival (OS) of 19.3 months. For patients with ≥very good partial response (VGPR), median PFS and OS improved to 9 months and 34 months, respectively. Achievement of MRD negativity in ≥VGPR did not further improve the outcome. A better performance status, younger age, longer time interval from initial MM diagnosis/initial ASCT to salvage ASCT and low-risk GEP70 were all associated with improved PFS and OS after salvage ASCT. Our results suggest a role for salvage ASCT in selected heavily pretreated and Daratumumab-refractory patients.
MiRNAs Correlate with HLA Expression in Uveal Melanoma: Both Up- and Downregulation Are Related to Monosomy 3
Authors: Souri Zahra , Wierenga Annemijn P. A. , Kiliç Emine , Brosens Erwin , Böhringer Stefan , Kroes Wilma G. M. , Verdijk Robert M. , van der Velden Pieter A. , Luyten Gregorius P. M. , Jager Martine J.
Published: August 2021

MicroRNAs are known to play a role in the regulation of inflammation. As a high HLA Class I expression is associated with a bad prognosis in UM, we set out to determine whether any miRNAs were related to a high HLA Class I expression and inflammation. We also determined whether such miRNAs were related to the UM’s genetic status. The expression of 125 miRNAs was determined in 64 primary UM from Leiden. Similarly, the mRNA expression of HLA-A, HLA-B, TAP1, BAP1, and immune cell markers was obtained. Expression levels of 24 of the 125 miRNAs correlated with expression of at least three out of four HLA Class I probes. Four miRNAs showed a positive correlation with HLA expression and infiltration with leukocytes, 20 a negative pattern. In the first group, high miRNA levels correlated with chromosome 3 loss/reduced BAP1 mRNA expression, in the second group low miRNA levels. The positive associations between miRNA-22 and miRNA-155 with HLA Class I were confirmed in the TCGA study and Rotterdam cohort, and with TAP1 in the Rotterdam data set; the negative associations between miRNA-125b2 and miRNA-211 and HLA-A, TAP1, and CD4 were confirmed in the Rotterdam set. We demonstrate two patterns: miRNAs can either be related to a high or a low HLA Class I/TAP1 expression and the presence of infiltrating lymphocytes and macrophages. However, both patterns were associated with chromosome 3/BAP1 status, which suggests a role for BAP1 loss in the regulation of HLA expression and inflammation in UM through miRNAs.
Genetic Profiling of a Cohort of Italian Patients with ACTH-Secreting Pituitary Tumors and Characterization of a Novel USP8 Gene Variant
Authors: Treppiedi Donatella , Barbieri Anna Maria , Di Muro Genesio , Marra Giusy , Mangili Federica , Catalano Rosa , Esposito Emanuela , Ferrante Emanuele , Serban Andreea Liliana , Locatelli Marco , Lania Andrea Gerardo , Spada Anna , Arosio Maura , Peverelli Erika , Mantovani Giovanna
Published: August 2021

Cushing’s Disease (CD) is a rare condition characterized by an overproduction of ACTH by an ACTH-secreting pituitary tumor, resulting in an excess of cortisol release by the adrenal glands. Somatic mutations in the deubiquitinases USP8 and USP48, and in BRAF genes, have been reported in a subset of patients affected by CD. The aim of this study was to characterize the genetic profile of a cohort of 60 patients with ACTH-secreting tumors, searching for somatic mutations in USP8, USP48, and BRAF hotspot regions. Seven patients were found to carry USP8 somatic mutations in the well-characterized 14-3-3 protein binding motif (n = 5 P720R, n = 1 P720Q, n = 1 S718del); 2 patients were mutated in USP48 (M415I); no mutation was identified in BRAF. In addition, a novel USP8 variant, G664R, located in exon 14, upstream of the 14-3-3 protein binding motif, was identified in 1 patient. Functional characterization of USP8 G664R variant was performed in murine corticotroph tumor AtT-20 cells. Transient transfection with the USP8 G664R variant resulted in a significant increase of ACTH release and cell proliferation (+114.5 ± 53.6% and +28.3 ± 2.6% vs. empty vector transfected cells, p < 0.05, respectively). Notably, USP8 proteolytic cleavage was enhanced in AtT-20 cells transfected with G664R USP8 (1.86 ± 0.58–fold increase of N-terminal USP8 fragment, vs. WT USP8, p < 0.05). Surprisingly, in situ Proximity Ligation Assay (PLA) experiments showed a significant reduction of PLA positive spots, indicating USP8/14-3-3 proteins colocalization, in G664R USP8 transfected cells with respect to WT USP8 transfected cells (−47.9 ± 6.6%, vs. WT USP8, p < 0.001). No significant difference in terms of ACTH secretion, cell proliferation and USP8 proteolytic cleavage, and 14-3-3 proteins interaction was observed between G664R USP8 and S718del USP8 transfected cells. Immunofluorescence experiments showed that, contrary to S718del USP8 but similarly to WT USP8 and other USP8 mutants, G664R USP8 displays an exclusive cytoplasmic localization. In conclusion, somatic mutations were found in USP8 (13.3% vs. 36.5% incidence of all published mutations) and USP48 (3.3% vs. 13.3% incidence) hotspot regions. A novel USP8 variant was identified in a CD patient, and in vitro functional studies in AtT-20 cells suggested that this somatic variant might be clinically relevant in ACTH-secreting tumor pathogenesis, expanding the characterization of USP8 functional domains.
Correction: Weng et al. New International Association for the Study of Lung Cancer (IASLC) Pathology Committee Grading System for the Prognostic Outcome of Advanced Lung Adenocarcinoma. Cancers 2020, 12, 3426
Authors: Weng Ching-Fu , Huang Chi-Jung , Huang Shih-Hung , Wu Mei-Hsuan , Tseng Ailun Heather , Sung Yung-Chuan , Lee Henry Hsin-Chung , Ling Thai-Yen
Published: August 2021

The authors would like to make a correction to their published paper [...]