bluuebunny/crossref_metadata_2025 huggingface dataset:
Cyclic Tetra-Adenylate (cA4) Recognition by Csa3; Implications for an Integrated Class 1 CRISPR-Cas Immune Response in Saccharolobus solfataricus
Authors: Charbonneau Alexander A. , Eckert Debra M. , Gauvin Colin C. , Lintner Nathanael G. , Lawrence C. Martin
Published: December 2021

Csa3 family transcription factors are ancillary CRISPR-associated proteins composed of N-terminal CARF domains and C-terminal winged helix-turn-helix domains. The activity of Csa3 transcription factors is thought to be controlled by cyclic oligoadenyate (cOA) second messengers produced by type III CRISPR-Cas surveillance complexes. Here we show that Saccharolobus solfataricus Csa3a recognizes cyclic tetra-adenylate (cA4) and that Csa3a lacks self-regulating “ring nuclease” activity present in some other CARF domain proteins. The crystal structure of the Csa3a/cA4 complex was also determined and the structural and thermodynamic basis for cA4 recognition are described, as are conformational changes in Csa3a associated with cA4 binding. We also characterized the effect of cA4 on recognition of putative DNA binding sites. Csa3a binds to putative promoter sequences in a nonspecific, cooperative and cA4-independent manner, suggesting a more complex mode of transcriptional regulation. We conclude the Csa3a/cA4 interaction represents a nexus between the type I and type III CRISPR-Cas systems present in S. solfataricus, and discuss the role of the Csa3/cA4 interaction in coordinating different arms of this integrated class 1 immune system to mount a synergistic, highly orchestrated immune response.
Characterization of miR-34a-Induced Epithelial–Mesenchymal Transition in Non-Small Lung Cancer Cells Focusing on p53
Authors: Kawami Masashi , Takenaka Shinnosuke , Akai Mizuki , Yumoto Ryoko , Takano Mikihisa
Published: December 2021

Background: Epithelial–mesenchymal transition (EMT), a phenotypic conversion of the epithelial to mesenchymal state, contributes to cancer progression. Currently, several microRNAs (miRNAs) are associated with EMT-mediated cancer progression, but the contribution of miR-34a to EMT in cancer cells remains controversial. The present study aimed to clarify the role of miR-34a in the EMT-related phenotypes of human non-small cell lung cancer (NSCLC) cell lines, A549 (p53 wild-type) and H1299 (p53-deficient). Methods: The miR-34a mimic and p53 small interfering RNA (siRNA) were transfected into the cells using Lipofectamine, and the obtained total RNA and cell lysates were used for real-time polymerase chain reaction and Western blotting analysis, respectively. Results: The introduction of the miR-34a mimic led to an increase in the mRNA and protein expression levels of α-smooth muscle actin (α-SMA), a mesenchymal marker gene, in A549, but not in H1299 cells. Additionally, miR-34a-induced the upregulation of p53 activity and migration was observed in A549, but not in H1299 cells. However, under the p53-knockdown condition, only α-SMA upregulation by miR-34a was abolished. Conclusion: These findings indicate a close relationship between p53 and miR-34a-induced EMT in p53-wild type NSCLC cells, which provides novel insights about the role of miR-34a in EMT-like phenotypic changes in NSCLC.
The New Pharmacological Chaperones PBXs Increase α-Galactosidase A Activity in Fabry Disease Cellular Models
Authors: Besada Pedro , Gallardo-Gómez María , Pérez-Márquez Tania , Patiño-Álvarez Lucía , Pantano Sergio , Silva-López Carlos , Terán Carmen , Arévalo-Gómez Ana , Ruz-Zafra Aurora , Fernández-Martín Julián , Ortolano Saida
Published: December 2021

Fabry disease is an X-linked multisystemic disorder caused by the impairment of lysosomal α-Galactosidase A, which leads to the progressive accumulation of glycosphingolipids and to defective lysosomal metabolism. Currently, Fabry disease is treated by enzyme replacement therapy or the orally administrated pharmacological chaperone Migalastat. Both therapeutic strategies present limitations, since enzyme replacement therapy has shown low half-life and bioavailability, while Migalastat is only approved for patients with specific mutations. The aim of this work was to assess the efficacy of PBX galactose analogues to stabilize α-Galactosidase A and therefore evaluate their potential use in Fabry patients with mutations that are not amenable to the treatment with Migalastat. We demonstrated that PBX compounds are safe and effective concerning stabilization of α-Galactosidase A in relevant cellular models of the disease, as assessed by enzymatic activity measurements, molecular modelling, and cell viability assays. This experimental evidence suggests that PBX compounds are promising candidates for the treatment of Fabry disease caused by mutations which affect the folding of α-Galactosidase A, even for GLA variants that are not amenable to the treatment with Migalastat.
Permittivity-Inspired Microwave Resonator-Based Biosensor Based on Integrated Passive Device Technology for Glucose Identification
Authors: Yue Wei , Kim Eun-Seong , Zhu Bao-Hua , Chen Jian , Liang Jun-Ge , Kim Nam-Young
Published: December 2021

In this study, we propose a high-performance resonator-based biosensor for mediator-free glucose identification. The biosensor is characterized by an air-bridge capacitor and fabricated via integrated passive device technology on gallium arsenide (GaAs) substrate. The exterior design of the structure is a spiral inductor with the air-bridge providing a sensitive surface, whereas the internal capacitor improves indicator performance. The sensing relies on repolarization and rearrangement of surface molecules, which are excited by the dropped sample at the microcosmic level, and the resonance performance variation corresponds to the difference in glucose concentration at the macroscopic level. The air-bridge capacitor in the modeled RLC circuit serves as a bio-recognition element to glucose concentration (εglucoseC0), generating resonant frequency shifts at 0.874 GHz and 1.244 GHz for concentrations of 25 mg/dL and 300 mg/dL compared to DI water, respectively. The proposed biosensor exhibits excellent sensitivity at 1.38 MHz per mg/dL with a wide detection range for glucose concentrations of 25–300 mg/dL and a low detection limit of 24.59 mg/dL. Additionally, the frequency shift and concentration are highly linear with a coefficient of determination of 0.98823. The response time is less than 3 s. We performed multiple experiments to verify that the surface morphology reveals no deterioration and chemical binding, thus validating the reusability and reliability of the proposed biosensor.
Political Hearts of Darkness: The Dark Triad as Predictors of Political Orientations and Interest in Politics
Authors: Bell Edward , Kowalski Christopher Marcin , Vernon Philip Anthony , Schermer Julie Aitken
Published: December 2021

Background: This study investigated the relationships between the Dark Triad of personality (sub-clinical psychopathy, Machiavellianism, and narcissism) and four political variables: socio-religious conservatism, support for greater economic equality, overall liberal–conservative orientation, and interest in politics. A theoretical approach that focused on the influence of the Dark Triad in large groups was provided to interpret those relationships. Methodological issues found in previous research that related to the use of abbreviated scales to measure the dark traits and the use of unidimensional indicators of political orientations were addressed. Methods: A hierarchical regression analysis was conducted to determine whether any of the three dark traits could explain variance in the aforementioned political attributes over and above that accounted for by the Big Five, sex, age, and nationality, using the full personality scales and measures of political orientation that captured both social and economic liberalism–conservatism. Results: Machiavellianism uniquely predicted lower levels of socio-religious conservatism, and both Machiavellianism and narcissism uniquely predicted lower levels of overall conservatism. Conclusions: There were important links between the Dark Triad and politics.
A Dual-Process Model Applied to Two Health-Promoting Nutrition Behaviours
Authors: Brown Daniel J. , Charlesworth Jessica , Hagger Martin S. , Hamilton Kyra
Published: December 2021

We tested a dual process model incorporating constructs that reflect both performing the target behaviour (behaviour directed habit) and habits that run counter to the target behaviour (opposing behaviour habit) in accounting for variance in two health behaviours: eating the recommended serves of fruits and vegetables a day and restricting sugar-sweetened beverage consumption. A prospective correlational design with two waves of data collection separated by one week was adopted. Participants (N = 606) comprising middle school students (n = 266) and university students (n = 340) completed an initial survey comprising self-report measures of past behaviour, intention, and habit to perform the target behaviour and habits that run counter to the target behaviour. One week later, participants (N = 414) completed a self-reported measure of behaviour. Results revealed that behaviour directed habits predicted fruit and vegetable consumption in both samples, while opposing behaviour habits predicted restriction of sugar-sweetened beverages in the middle-school sample only, with a moderating effect also observed. Current findings indicate that habits specifying avoidance of the target behaviour did not predict future behaviour. However, the moderating effect observed provides preliminary evidence that strong habits to perform a behaviour may override habit to avoid the behaviour.
Communication Styles and Attention Performance in Primary School Children
Authors: de la Torre Gabriel G. , Ramallo Miguel A. , Gonzalez-Torre Sara , Mora Prat Alvaro , Rueda-Marroquin Andrea , Sallago-Marcos Amanda , Toro-Barrios Zoraida , Garcia Manuel A.
Published: December 2021

Communication styles are the three communication patterns that result from merging the verbal elements of communication, the non-verbal elements and the paraverbal elements. The objective of this study was to test what effect different communication styles have on attention performance in primary school children. We administered the assertive behavior scale for children (CABS), the symbol digit modalities test (SDMT) and the d2 attention test to a sample of 77 participants of primary school. A statistically significant and proportional correlation was found between the assertive communication style and the total number of correct answers of the SDMT. We also found positive correlations between the assertive communication style and d2 attention test performance. Children with an assertive communication style have better attention performance than children with passive or aggressive communication styles.
Project Data Categorization, Adoption Factors, and Non-Functional Requirements for Blockchain Based Digital Twins in the Construction Industry 4.0
Authors: Teisserenc Benjamin , Sepasgozar Samad
Published: December 2021

As key technologies of the fourth industrial revolution, blockchain and digital twins have great potential to enhance collaboration, data sharing, efficiency, and sustainability in the construction industry. Blockchain can improve data integrity and enhance trust in the data value chain throughout the entire lifecycle of projects. This paper aims to develop a novel theoretical framework for the adoption of environmentally sustainable blockchain-based digital twins (BCDT) for Construction Industry (CI) 4.0. The paper identifies which key data from construction projects lifecycle should be anchored in BCDTs to benefit CI 4.0 and the environment. The paper also identifies key factors and non-functional requirements necessary for the adoption of BCDTs in a decentralized and sustainable CI 4.0. At first, a content analysis of the literature allowed the identification of which data from projects lifecycle would benefit from blockchain technology (BCT) adoption and what the key factors and non-functional requirements necessary for the adoption of BCDT in the CI4.0 are. Furthermore, the analysis of structured interviews and online survey permitted to firstly validate the hypotheses raised from the literature and to offer a novel framework for BCDT of CI 4.0 in the context of the circular economy (CE). The findings are that (1) the key project lifecycle data relevant for BCDTs relate to the BIM dimensions (3D, 4D, 5D, 6D, 7D, and 8D) and a new dimension called the contractual dimension (cD) is also proposed. (2) Ecosystems of BCDTs should embrace a novel form of collaboration that is decentralized and presented as Level 4 maturity for BCDTs. This new level of maturity leverages distributed blockchain networks to enhance collaboration, processes automation with smart contracts, and data sharing within a decentralized data value chain. Finally (3), the main non-functional requirements for BCDTs are security, privacy, interoperability, data ownership, data integrity, and the decentralization and scalability of data storage. With the proposed framework including the BCDT dimensions, the Maturity Level 4, and the key non-functional requirements, this paper provides the building blocks for industry practitioners to adopt BCDTs. This is promising for CI 4.0 to embrace a paradigm shift towards decentralized ecosystems of united BCDTs where trust, collaboration, data sharing, information security, efficiency, and sustainability are improved throughout the lifecycle of projects and within a decentralized CE (DCE).
Research Progress of Urban Floods under Climate Change and Urbanization: A Scientometric Analysis
Authors: Yang Qiu , Zheng Xiazhong , Jin Lianghai , Lei Xiaohui , Shao Bo , Chen Yun
Published: December 2021

Urban floods research has been attracting extensive attention with the increasing threat of flood risk and environmental hazards due to global climate change and urbanization. However, there is rarely a comprehensive review of this field and it remains unclear how the research topics on urban floods have evolved. In this study, we analyzed the development of urban floods research and explored the hotspots and frontiers of this field by scientific knowledge mapping. In total, 3314 published articles from 2006 to 2021 were analyzed. The results suggest that the number of published articles in the field of urban floods generally has an upward trend year by year, and the research focus has shifted from exploring hydrological processes to adopting advanced management measures to solve urban flood problems. Moreover, urban stormwater management and low impact development in the context of climate change and urbanization have gradually become research hotspots. Future research directions based on the status and trends of the urban floods field were also discussed. This research can not only inspire other researchers and policymakers, but also demonstrates the effectiveness of scientific knowledge mapping analysis by the use of the software CiteSpace and VOSviewer.
A Scientometric Analysis and Overview of Research on Infrastructure Externalities
Authors: Zhu Lei , Ye Qianwen , Yuan Jingfeng , Hwang Bon-Gang , Cheng Yusi
Published: December 2021

There is a growing level of concern regarding infrastructure externalities globally. However, most of the previous relevant reviews were undertaken manually and few of them covered all infrastructure systems. This study conducted a scientometric analysis and overview of the research on externalities of all infrastructure systems. The analysis results of 743 articles that were rigorously selected first showed the increasing trend of research interest in infrastructure externalities. Moreover, the results demonstrated productive and influential journals, scholars, and institutions, and their collaboration networks. Furthermore, research on the spillover effects of the infrastructure on economic growth, airport-related externalities, road transport-related externalities, and externalities of ecosystem services and energy systems were identified as the four main research domains. The evolution of the research is reflected in the focus change from economic aspects to environment aspects, from government governance to assessing and pricing by the market, and from airport to other infrastructure systems. Additionally, this study identified the scientific knowledge base supporting each research domain. Finally, this study pointed out research gaps and future research directions in the aspects of knowledge base, multi-dimensional evaluation, and multiple governance strategies. The results could cultivate deeper and more carefully focused research into this field in the academic community, and assist policymakers and practitioners in research planning and funding efforts.
From Audits to Projects: Evaluating New York State Policy to Encourage Home Retrofit Projects
Authors: Hewitt Elizabeth , Boucher Jean
Published: December 2021

This research explores socio-spatial characteristics of home retrofit projects in New York State and their association with the state’s free home energy audit program. Prior work by the authors found that zip codes with more elder individuals and higher levels of education are more likely to undertake an energy audit; it was also found that higher incomes may be negatively correlated with audit decisions. Less understood is the follow-up decision after an audit to undertake a retrofit. From a policy and climate perspective, the actionable retrofit decision is far more impactful than the informational audit, making it an important area of further research. This work examines this understudied area using a combination of datasets, including census data, American Community Survey (ACS) data, and retrofits data provided by the New York State Energy Research and Development Authority (NYSERDA). Findings indicate that many of the same socioeconomic characteristics that predict audits are influential in retrofit projects as well (age, education, higher home values). A strong statistical relationship was found between audits per capita and subsequent retrofit projects, which is to be expected, as NYSERDA requires audits of residents desiring efficiency retrofits. However, this also indicates that the role of the audit in information transfers may be highly influential in encouraging home energy efficiency projects. This finding underscores the policy importance of offering low- or no-cost energy audit incentives to encourage greater participation in home retrofit programs.
Evaluation of the Thermal Comfort and Energy Demand in a Building with Rammed Earth Walls in Spain: Influence of the Use of In Situ Measured Thermal Conductivity and Estimated Values
Authors: Mellado Mascaraque Miguel Ángel , Castilla Pascual Francisco Javier , Pérez Andreu Víctor , Gosalbo Guenot Guillermo Adrián
Published: December 2021

This paper describes the influence of thermal parameters—conductivity, transmittance, and thermal mass—in the estimation of comfort and energy demand of a building with rammed earth walls, and consequently, the compliance with standards. It is known that nominal design data does not match in situ measured values, especially in traditionally constructed buildings. We have therefore monitored a room in a building with rammed earth walls, designed a computerised model, and compared four different alternatives where we have changed the value for the thermal conductivity (in situ vs. estimated) and the consideration of thermal mass. When we then analyse the compliance with the Spanish energy saving code, using measured values would result in lower differences with the standards’ limits and even comply with the global thermal transmittance (K-value) requirement. This would mean a more realistic approach to the restoration of traditional buildings leading to the use of thinner and more suitable insulation and retrofitting systems, encouraging the use of rammed earth in new buildings, and therefore reducing the carbon footprint due to materials used in construction. Results show that the building model that uses in situ values and considers thermal mass (S1) is closer to reality when assessing thermal comfort. Finally, using nominal data would result in requiring 43% more energy in the selected winter period and 102% more energy in the selected summer period to keep the same comfort conditions as in the alternative where measured values are used.
Advances in Biomarker-Driven Targeted Therapies in Thyroid Cancer
Authors: Mishra Prachi , Laha Dipranjan , Grant Robert , Nilubol Naris
Published: December 2021

Thyroid cancer is the most common type of endocrine malignancy comprising 2–3% of all cancers, with a constant rise in the incidence rate. The standard first-line treatments for thyroid cancer include surgery and radioactive iodine ablation, and a majority of patients show a good response to these therapies. Despite a better response and outcome, approximately twenty percent of patients develop disease recurrence and distant metastasis. With improved knowledge of molecular dysregulation and biological characteristics of thyroid cancer, the development of new treatment strategies comprising novel targets has accelerated. Biomarker-driven targeted therapies have now emerged as a trend for personalized treatments in patients with advanced cancers, and several multiple receptor kinase inhibitors have entered clinical trials (phase I/II/III) to evaluate their safety and efficacy. Most extensively investigated and clinically approved targeted therapies in thyroid cancer include the tyrosine receptor kinase inhibitors that target antiangiogenic markers, BRAF mutation, PI3K/AKT, and MAPK pathway components. In this review, we focus on the current advances in targeted mono- and combination therapies for various types of thyroid cancer.
Fine Crystalline Mg-Al Hydrotalcites as Catalysts for Baeyer-Villiger Oxidation of Cyclohexanone with H2O2
Authors: Karcz Robert , Napruszewska Bogna D. , Michalik Alicja , Kryściak-Czerwenka Joanna , Duraczyńska Dorota , Serwicka Ewa M.
Published: December 2021

The catalytic activity of Mg-Al hydrotalcite (HT) materials in base-catalyzed reactions is known to be promoted by the low crystallinity of the HT solid. In the present work, two routes enabling the preparation of finely crystalline Mg-Al HT materials were explored: (1) the inverse microemulsion technique, and (2) co-precipitation in the presence of starch. Carbonate, chloride and bromide forms of HT were prepared, examined with X-ray diffraction, scanning electron microscopy/energy dispersive X-ray spectroscopy and infrared spectroscopy, and used as catalysts in the Baeyer–Villiger oxidation of cyclohexanone to ε-caprolactone with a H2O2/acetonitrile system. The bromide forms proved significantly less active than the chlorides and carbonates, as they promoted nonselective consumption of H2O2. The fine crystalline materials were more active than the more crystalline HT references obtained by conventional co-precipitation. Catalysts prepared by inverse microemulsion were less crystalline and more active than the starch-templated ones, but suffered stronger deactivation by the acidic reaction environment. Alkalization of the reaction medium with NaHCO3 stabilized the HT materials and increased the ε-caprolactone yield, which became comparable for both types of fine crystalline catalysts—thus pointing to the synthesis involving a simple and cheap starch templating approach as being a particularly attractive one.
Synthesis of Indigo-Dyes from Indole Derivatives by Unspecific Peroxygenases and Their Application for In-Situ Dyeing
Authors: Ullrich René , Poraj-Kobielska Marzena , Herold-Majumdar Owik M. , Vind Jesper , Hofrichter Martin
Published: December 2021

Tyrian purple (also known as royal or imperial purple) is the oldest known commercial pigment and still one of the most expensive dyes, often associated with the wardrobes of clergy and royalty. It is a brominated derivative of indigo, a natural dye that has been used since 4000 BC. Moreover, just recently, the therapeutic value of indigoids for the treatment of several disorders was discovered. The manufacturing of indigo derivatives by the existing chemical routes has become increasingly uninteresting due to the use of aggressive reagents, expensive starting materials and high-energy costs. Thus, both dyestuff manufacturers and the pharmaceutical industry are interested in the development of gentle preparation methods of indigoids from simple precursors. Here, we describe a simple enzymatic method for the one-step synthesis of Tyrian purple and other indigo derivatives with fungal peroxygenases (UPO, EC 1.11.2.1). The reaction does not require complex co-substrates and works well in phosphate buffers with H2O2 (<0.1 wt%) and less than 5% (v/v) acetonitrile as co-solvent. We demonstrate the scaling up of the reaction to 10 Liters and established thereupon an environmentally friendly combined synthesis and in-situ dyeing process, further simplifying the manufacturing of vat-dyed fabrics. Eventually, we screened a number of halogen-substituted indoles in the search for novel indigo derivatives, which may be of interest for pharmaceutical and/or dyeing purposes.
Defective Grey TiO2 with Minuscule Anatase–Rutile Heterophase Junctions for Hydroxyl Radicals Formation in a Visible Light-Triggered Photocatalysis
Authors: Ullattil Sanjay Gopal , Zavašnik Janez , Maver Ksenija , Finšgar Matjaž , Novak Tušar Nataša , Pintar Albin
Published: December 2021

The novelty of this work was to prepare a series of defect-rich colored TiO2 nanostructures, using a peroxo solvothermal-assisted, high-pressure nitrogenation method. Among these solids, certain TiO2 materials possessed a trace quantity of anatase–rutile heterojunctions, which are beneficial in obtaining high reaction rates in photocatalytic reactions. In addition, high surface area (above 100 m2/g), even when utilizing a high calcination temperature (500 °C), and absorption of light at higher wavelengths, due to the grey color of the synthesized titania, were observed as an added advantage for photocatalytic hydroxyl radical formation. In this work, we adopted a photoluminescent probe method to monitor the temporal evolution of hydroxyl radicals. As a result, promising hydroxyl radical formations were observed for all the colored samples synthesized at 400 and 500 °C, irrespective of the duration of calcination.
Transaminase Catalysis for Enantiopure Saturated Heterocycles as Potential Drug Scaffolds
Authors: Malta-Lakó Ágnes , Zhang Fangyi , Mendonça Ricardo , Poppe László
Published: December 2021

As efforts in rational drug design are driving the pharmaceutical industry towards more complex molecules, the synthesis and production of these new drugs can benefit from new reaction routes. In addition to the introduction of new centers of asymmetry, complexity can be also increased by ring saturation, which also provides improved developability measures. Therefore, in this report, our aim was to develop transaminase (TA)-catalyzed asymmetric synthesis of a new group of potential chiral drug scaffolds comprising a saturated amine heterocycle backbone and an asymmetric primary amine sidechain (55a–g). We screened the Codex® Amine Transaminase Kit of 24 transaminases with the morpholine containing ketone 57a, resulting in one (R)-selective TA and three (S)-selective TAs operating at 100 mM substrate concentration and 25 v/v% isopropylamine (IPA) content. The optimized reaction conditions were than applied for asymmetric transamination of further six ketones (57b–g) containing various amine heterocycles, in which a strong effect of the substitution pattern of the γ-position relative to the substituted N-atom could be observed. Mediated by the most enantiotope selective (S)-TAs in scaled-up process, the (S)-amines [(S)-55a–g] were isolated with moderate-to-excellent yields (47–94%) in enantiopure form (>99% ee).
Engineered Stable 5-Hydroxymethylfurfural Oxidase (HMFO) from 8BxHMFO Variant of Methylovorus sp. MP688 through B-Factor Analysis
Authors: Wu Qiuyang , Lu Dong , Jin Shuming , Lu Jie , Wang Fang , Liu Luo , Nie Kaili
Published: December 2021

What is known as Furan-2,5-dicarboxylic acid (FDCA) is an attractive compound since it has similar properties to terephthalic acid. Further, 5-hydroxymethylfurfural oxidase (HMFO) is an enzyme, which could convert HMF to FDCA directly. Most wild types of HMFO have low activity on the oxidation of HMF to FDCA. The variant of 8BxHFMO from Methylovorus sp. MP688 was the only reported enzyme that was able to perform FDCA production. However, the stabilization of 8BxHMFO is still not that satisfactory, and further improvement is necessary for the industrial application of the enzyme. In this work, stability-enhanced HMFO from 8BxHFMO was engineered through employing B-factor analysis. The mutation libraries were created based on the NNK degeneracy of residues with the top ten highest B-factor value, and two of the effective mutants were screened out through the high throughput selection with the horseradish peroxidase (HRP)-Tyr assay. The mutants Q319K and N44G show a significantly increased yield of FDCA in the reaction temperature range of 30 to 40 °C. The mutant Q319K shows the best performance at 35 °C with a FDCA yield of 98% (the original 8BxHMFO was only 85%), and a half-life exceeding 72 h. Moreover, molecular dynamic simulation indicates that more hydrogen bonds are formed in the mutants, which improves the stability of the protein structure. The method could enhance the design of more stable biocatalysts; and provides potential for the further optimization and utilization of HMFO in biotechnological processes.
Synthesis and Performance of Photocatalysts for Photocatalytic Hydrogen Production: Future Perspectives
Authors: Escobedo Salvador , de Lasa Hugo
Published: December 2021

Photocatalysis for “green” hydrogen production is a technology of increasing importance that has been studied using both TiO2–based and heterojunction composite-based semiconductors. Different irradiation sources and reactor units can be considered for the enhancement of photocatalysis. Current approaches also consider the use of electron/hole scavengers, organic species, such as ethanol, that are “available” in agricultural waste, in communities around the world. Alternatively, organic pollutants present in wastewaters can be used as organic scavengers, reducing health and environmental concerns for plants, animals, and humans. Thus, photocatalysis may help reduce the carbon footprint of energy production by generating H2, a friendly energy carrier, and by minimizing water contamination. This review discusses the most up-to-date and important information on photocatalysis for hydrogen production, providing a critical evaluation of: (1) The synthesis and characterization of semiconductor materials; (2) The design of photocatalytic reactors; (3) The reaction engineering of photocatalysis; (4) Photocatalysis energy efficiencies; and (5) The future opportunities for photocatalysis using artificial intelligence. Overall, this review describes the state-of-the-art of TiO2–based and heterojunction composite-based semiconductors that produce H2 from aqueous systems, demonstrating the viability of photocatalysis for “green” hydrogen production.
White Adipose Tissue Depots Respond to Chronic Beta-3 Adrenergic Receptor Activation in a Sexually Dimorphic and Depot Divergent Manner
Authors: Queathem Eric D. , Welly Rebecca J. , Clart Laura M. , Rowles Candace C. , Timmons Hunter , Fitzgerald Maggie , Eichen Peggy A. , Lubahn Dennis B. , Vieira-Potter Victoria J.
Published: December 2021

Beta-3 adrenergic receptor activation via exercise or CL316,243 (CL) induces white adipose tissue (WAT) browning, improves glucose tolerance, and reduces visceral adiposity. Our aim was to determine if sex or adipose tissue depot differences exist in response to CL. Daily CL injections were administered to diet-induced obese male and female mice for two weeks, creating four groups: male control, male CL, female control, and female CL. These groups were compared to determine the main and interaction effects of sex (S), CL treatment (T), and WAT depot (D). Glucose tolerance, body composition, and energy intake and expenditure were assessed, along with perigonadal (PGAT) and subcutaneous (SQAT) WAT gene and protein expression. CL consistently improved glucose tolerance and body composition. Female PGAT had greater protein expression of the mitochondrial uncoupling protein 1 (UCP1), while SQAT (S, p < 0.001) was more responsive to CL in increasing UCP1 (S×T, p = 0.011) and the mitochondrial biogenesis induction protein, PPARγ coactivator 1α (PGC1α) (S×T, p = 0.026). Females also displayed greater mitochondrial OXPHOS (S, p < 0.05) and adiponectin protein content (S, p < 0.05). On the other hand, male SQAT was more responsive to CL in increasing protein levels of PGC1α (S×T, p = 0.046) and adiponectin (S, p < 0.05). In both depots and in both sexes, CL significantly increased estrogen receptor beta (ERβ) and glucose-related protein 75 (GRP75) protein content (T, p < 0.05). Thus, CL improves systemic and adipose tissue-specific metabolism in both sexes; however, sex differences exist in the WAT-specific effects of CL. Furthermore, across sexes and depots, CL affects estrogen signaling by upregulating ERβ.
Chromatin-Independent Interplay of NFATc1 and EZH2 in Pancreatic Cancer
Authors: Patil Shilpa , Forster Teresa , Reutlinger Kristina , Kopp Waltraut , Versemann Lennart , Spitalieri Jessica , Gaedcke Jochen , Ströbel Philipp , Singh Shiv K. , Ellenrieder Volker , Neesse Albrecht , Hessmann Elisabeth
Published: December 2021

Background: The Nuclear Factor of Activated T-cells 1 (NFATc1) transcription factor and the methyltransferase Enhancer of Zeste Homolog 2 (EZH2) significantly contribute to the aggressive phenotype of pancreatic ductal adenocarcinoma (PDAC). Herein, we aimed at dissecting the mechanistic background of their interplay in PDAC progression. Methods: NFATc1 and EZH2 mRNA and protein expression and complex formation were determined in transgenic PDAC models and human PDAC specimens. NFATc1 binding on the Ezh2 gene and the consequences of perturbed NFATc1 expression on Ezh2 transcription were explored by Chromatin Immunoprecipitation (ChIP) and upon transgenic or siRNA-mediated interference with NFATc1 expression, respectively. Integrative analyses of RNA- and ChIP-seq data was performed to explore NFATc1-/EZH2-dependent gene signatures. Results: NFATc1 targets the Ezh2 gene for transcriptional activation and biochemically interacts with the methyltransferase in murine and human PDAC. Surprisingly, our genome-wide binding and expression analyses do not link the protein complex to joint gene regulation. In contrast, our findings provide evidence for chromatin-independent functions of the NFATc1:EZH2 complex and reveal posttranslational EZH2 phosphorylation at serine 21 as a prerequisite for robust complex formation. Conclusion: Our findings disclose a previously unknown NFATc1-EZH2 axis operational in the pancreas and provide mechanistic insights into the conditions fostering NFATc1:EZH2 complex formation in PDAC.
Characterization of Extracellular Vesicles from Bronchoalveolar Lavage Fluid and Plasma of Patients with Lung Lesions Using Fluorescence Nanoparticle Tracking Analysis
Authors: Dlugolecka Magdalena , Szymanski Jacek , Zareba Lukasz , Homoncik Zuzanna , Domagala-Kulawik Joanna , Polubiec-Kownacka Malgorzata , Czystowska-Kuzmicz Malgorzata
Published: December 2021

The current lack of reliable methods for quantifying extracellular vesicles (EVs) isolated from complex biofluids significantly hinders translational applications in EV research. The recently developed fluorescence nanoparticle tracking analysis (FL-NTA) allows for the detection of EV-associated proteins, enabling EV content determination. In this study, we present the first comprehensive phenotyping of bronchopulmonary lavage fluid (BALF)-derived EVs from non-small cell lung cancer (NSCLC) patients using classical EV-characterization methods as well as the FL-NTA method. We found that EV immunolabeling for the specific EV marker combined with the use of the fluorescent mode NTA analysis can provide the concentration, size, distribution, and surface phenotype of EVs in a heterogeneous solution. However, by performing FL-NTA analysis of BALF-derived EVs in comparison to plasma-derived EVs, we reveal the limitations of this method, which is suitable only for relatively pure EV isolates. For more complex fluids such as plasma, this method appears to not be sensitive enough and the measurements can be compromised. Our parallel presentation of NTA-based phenotyping of plasma and BALF EVs emphasizes the great impact of sample composition and purity on FL-NTA analysis that has to be taken into account in the further development of FL-NTA toward the detection of EV-associated cancer biomarkers.
Phenotypical and Myopathological Consequences of Compound Heterozygous Missense and Nonsense Variants in SLC18A3
Authors: Della Marina Adela , Arlt Annabelle , Schara-Schmidt Ulrike , Depienne Christel , Gangfuß Andrea , Kölbel Heike , Sickmann Albert , Freier Erik , Kohlschmidt Nicolai , Hentschel Andreas , Weis Joachim , Czech Artur , Grüneboom Anika , Roos Andreas
Published: December 2021

Background: Presynaptic forms of congenital myasthenic syndromes (CMS) due to pathogenic variants in SLC18A3 impairing the synthesis and recycling of acetylcholine (ACh) have recently been described. SLC18A3 encodes the vesicular ACh transporter (VAChT), modulating the active transport of ACh at the neuromuscular junction, and homozygous loss of VAChT leads to lethality. Methods: Exome sequencing (ES) was carried out to identify the molecular genetic cause of the disease in a 5-year-old male patient and histological, immunofluorescence as well as electron- and CARS-microscopic studies were performed to delineate the muscle pathology, which has so far only been studied in VAChT-deficient animal models. Results: ES unraveled compound heterozygous missense and nonsense variants (c.315G>A, p.Trp105* and c.1192G>C, p.Asp398His) in SLC18A3. Comparison with already-published cases suggests a more severe phenotype including impaired motor and cognitive development, possibly related to a more severe effect of the nonsense variant. Therapy with pyridostigmine was only partially effective while 3,4 diaminopyridine showed no effect. Microscopic investigation of the muscle biopsy revealed reduced fibre size and a significant accumulation of lipid droplets. Conclusions: We suggest that nonsense variants have a more detrimental impact on the clinical manifestation of SLC18A3-associated CMS. The impact of pathogenic SLC18A3 variants on muscle fibre integrity beyond the effect of denervation is suggested by the build-up of lipid aggregates. This in turn implicates the importance of proper VAChT-mediated synthesis and recycling of ACh for lipid homeostasis in muscle cells. This hypothesis is further supported by the pathological observations obtained in previously published VAChT-animal models.
Human Induced Pluripotent Stem Cell as a Disease Modeling and Drug Development Platform—A Cardiac Perspective
Authors: Bekhite Mohamed M. , Schulze P. Christian
Published: December 2021

A comprehensive understanding of the pathophysiology and cellular responses to drugs in human heart disease is limited by species differences between humans and experimental animals. In addition, isolation of human cardiomyocytes (CMs) is complicated because cells obtained by biopsy do not proliferate to provide sufficient numbers of cells for preclinical studies in vitro. Interestingly, the discovery of human-induced pluripotent stem cell (hiPSC) has opened up the possibility of generating and studying heart disease in a culture dish. The combination of reprogramming and genome editing technologies to generate a broad spectrum of human heart diseases in vitro offers a great opportunity to elucidate gene function and mechanisms. However, to exploit the potential applications of hiPSC-derived-CMs for drug testing and studying adult-onset cardiac disease, a full functional characterization of maturation and metabolic traits is required. In this review, we focus on methods to reprogram somatic cells into hiPSC and the solutions for overcome immaturity of the hiPSC-derived-CMs to mimic the structure and physiological properties of the adult human CMs to accurately model disease and test drug safety. Finally, we discuss how to improve the culture, differentiation, and purification of CMs to obtain sufficient numbers of desired types of hiPSC-derived-CMs for disease modeling and drug development platform.
The Interleukins Orchestrate Mucosal Immune Responses to Salmonella Infection in the Intestine
Authors: Huang Fu-Chen
Published: December 2021

Salmonella infection remains one of the major public health problems in the world, with increasing resistance to antibiotics. The resolution is to explore the pathogenesis of the infection and search for alternative therapy other than antibiotics. Immune responses to Salmonella infection include innate and adaptive immunity. Flagellin or muramyl dipeptide from Salmonella, recognized by extracellular Toll-like receptors and intracellular nucleotide-binding oligomerization domain2, respectively, induce innate immunity involving intestinal epithelial cells, neutrophils, macrophages, dendric cells and lymphocytes, including natural killer (NK) and natural killer T (NKT) cells. The cytokines, mostly interleukins, produced by the cells involved in innate immunity, stimulate adaptive immunity involving T and B cells. The mucosal epithelium responds to intestinal pathogens through its secretion of inflammatory cytokines, chemokines, and antimicrobial peptides. Chemokines, such as IL-8 and IL-17, recruit neutrophils into the cecal mucosa to defend against the invasion of Salmonella, but induce excessive inflammation contributing to colitis. Some of the interleukins have anti-inflammatory effects, such as IL-10, while others have pro-inflammatory effects, such as IL-1β, IL-12/IL-23, IL-15, IL-18, and IL-22. Furthermore, some interleukins, such as IL-6 and IL-27, exhibit both pro- and anti-inflammatory functions and anti-microbial defenses. The majority of interleukins secreted by macrophages and lymphocytes contributes antimicrobial defense or protective effects, but IL-8 and IL-10 may promote systemic Salmonella infection. In this article, we review the interleukins involved in Salmonella infection in the literature.
Is Drought Increasing in Maine and Hurting Wild Blueberry Production?
Authors: Barai Kallol , Tasnim Rafa , Hall Bruce , Rahimzadeh-Bajgiran Parinaz , Zhang Yong-Jiang
Published: December 2021

A few severe drought events occurred in the Northeast (NE) USA in recent decades and caused significant economic losses, but the temporal pattern of drought incidents and their impacts on agricultural systems have not been well assessed. Here, we analyzed historical changes and patterns of drought using a drought index (standardized precipitation-evapotranspiration index (SPEI)), and assessed drought impacts on remotely sensed vegetation indices (enhanced vegetation index (EVI) and normalized difference vegetation index (NDVI)) and production (yield) of the wild blueberry fields in Maine, USA. We also analyzed the impact of short- and long-term water conditions of the growing season on the wild blueberry vegetation condition and production. No significant changes in the SPEI were found in the past 71 years, despite a significant warming pattern. There was also a significant relationship between the relatively long-term SPEI and the vegetation indices (EVI and NDVI), but not the short-term SPEI (one year). This suggests that the crop vigor of wild blueberries is probably determined by water conditions over a relatively long term. There were also significant relationships between 1-year water conditions (SPEI) and yield for a non-irrigated field, and between 4-year-average SPEI and the yield of all fields in Maine. The vegetation indices (EVI and NDVI) are not good predictors of wild blueberry yield, possibly because wild blueberry yield does not only depend on crop vigor, but also on other important variables such as pollination. We also compared an irrigated and a non-irrigated wild blueberry field at the same location (Deblois, Maine) where we found that irrigation decoupled the relationship between the SPEI and NDVI or EVI.
Design and Fabrication of Laser Protective Lenses Based on Multilayered Notch Filter with Low Residual Stress and Low Surface Roughness
Authors: Tien Chuen-Lin , Lin Hong-Yi , Cheng Kuan-Sheng , Chang Chih-Kai
Published: December 2021

We present a new laser protective lens based on a multilayered notch filter design with low residual stress and low surface roughness. An18-layer notch filter was prepared by electron beam evaporation with an ion-assisted deposition technique, which was composed of SiO2 and Nb2O5 with a center wavelength of 532 nm. The optical transmittance, residual stress, surface roughness, and surface morphology were measured by a UV/VIS/NIR spectrophotometer, Twyman–Green interferometer, scanning probe microscope, Linnik microscopic interferometer, and field-emission scanning electron microscopy (FE-SEM). The transmittance of the notch filters at center wavelength is 0.2%, and the average transmittance of the transmission band is about 70%. The residual stress of the notch filter is −0.298 GPa, and the root mean square surface roughness is 1.88 nm. The experimental results show that the optical transmittance meets the design requirements.
Microstructure of Ag Nano Paste Joint and Its Influence on Reliability
Authors: Yang Dongsheng , Huang Yilong , Tian Yanhong
Published: December 2021

In this paper, the microstructure of Ag nano paste joint was investigated in pressure-less sintering conditions, and the influence of the microstructure on the joint’s reliability was studied. Firstly, silver nanoparticles (Ag NPs) were synthesized using the redox reaction method. To tightly stack the Ag NPs in nano paste, Ag NPs with sizes of 30~50 nm and submicron-sized Ag particles were mixed. It was found that increasing the sintering temperature or sintering time can reduce the porosity of the bonding layer and the interfacial crack simultaneously, resulting in higher shear strength. When sintering at a temperature of 250 °C, a complete bonding interface was formed, with a 0.68 μm interdiffusion layer. At a higher temperature (300 °C), the bonding interface reached 1.5 μm, providing 35.9 ± 1.7 MPa of shear strength. The reliability of the die attachment was analyzed under thermal shocking from −65 °C to 150 °C for 50 cycles. As the crack could quickly grow through the interfacial defects, the separation ratio was 85% and 67% when sintered at 150 °C and 200 °C, respectively. Because of the reliable bonding interface between the die and the substrate, the Ag nano paste joint formed a slight crack on the edge of the die when sintering at 250 °C. When the joint was sintered at 300 °C, the small voids became large voids, which featured lower resistance to crack growth. Thus, instead of further improved reliability, the separation ratio increased to 37%.