bluuebunny/crossref_metadata_2025 huggingface dataset:
Carbon Inequality Embodied in Inter-Provincial Trade of China’s Yangtze River Economic Belt
Authors: Ban Qingqing , Li Yiwen , Tian Guiliang , Wu Zheng , Xia Qing
Published: June 2023

Regional trade leads to carbon transfer, which in turn raises the issue of carbon inequality. However, less existing research has focused on carbon inequality within megaregions. Combining multi-regional input-output analysis, carbon Gini coefficients and carbon deviation coefficients, this paper presents a comprehensive analysis of carbon equity in inter-provincial trade in the Yangtze River Economic Belt (YEB) from the perspective of economic benefits and environmental pressure. The results show that: (1) Carbon emissions from the production and consumption sides of the 11 provinces in the YEB vary considerably. (2) Significant carbon inequality exists in the YEB region. This is manifested in the unequal relationship between the transfer of embodied carbon emissions and economic benefits, as well as the difference in carbon deviation coefficients. Based on the results of our research, this paper can help provide theoretical support and decision-making reference for implementing a differentiated carbon emission reduction responsibility mechanism and establishing a coordinated carbon emission reduction responsibility system.
Analyzing Independent LFMC Empirical Models in the Mid-Mediterranean Region of Spain Attending to Vegetation Types and Bioclimatic Zones
Authors: Arcos María Alicia , Edo-Botella Roberto , Balaguer-Beser Ángel , Ruiz Luis Ángel
Published: June 2023

This paper presents empirical models developed through stepwise multiple linear regression to estimate the live fuel moisture content (LFMC) in a Mediterranean area. The models are based on LFMC data measured in 50 field plots, considering four groups with similar bioclimatic characteristics and vegetation types (trees and shrubs). We also applied a species-specific LFMC model for Rosmarinus officinalis in plots with this dominant species. Spectral indices extracted from Sentinel-2 images and their averages over the study time period in each plot with a spatial resolution of 10 m were used as predictors, together with interpolated meteorological, topographic, and seasonal variables. The models achieved adjusted R2 values ranging between 52.1% and 74.4%. Spatial and temporal variations of LFMC in shrub areas were represented on a map. The results highlight the feasibility of developing satellite-derived LFMC operational empirical models in areas with various vegetation types and taking into account bioclimatic zones. The adjustment of data through GAM (generalized additive models) is also addressed in this study. The different error metrics obtained reflect that these models provided a better fit (most adjusted R2 values ranged between 65% and 74.1%) than the linear models, due to GAMs being more versatile and suitable for addressing complex problems such as LFMC behavior.
Digital Banks in Brazil: Struggling to Reach the Breakeven Point or a New Evolution Wave?
Authors: Bueno Luiz Antonio , Sigahi Tiago F. A. C. , Anholon Rosley
Published: June 2023

Digital banks have profoundly changed the financial industry’s operations. In this scenario, the study of digital banks has gained increasing attention in the academic community. However, there is still a lot of room to understand how this type of organization functions and impacts different contexts. Considering the information collected, partial findings, and the professional experience of those involved in a larger research project, the main objective of this study is to present the Brazilian scenario related to digital banks from the analytical perspective of the research group. The methodological approach included analysis of partial results of a larger research project, bibliographic research, analysis of public data about digital banks in Brazil, and multidisciplinary discursive approach to conduct debates with the support of academic literature and experience from top managers working in major Brazilian financial institutions. Data on key performance indicators (KPIs), including cost breakdown, net revenue, return on equity (ROE), and cost-to-income ratio, are presented and analyzed for both traditional and digital banks. Furthermore, this study puts forward potential avenues for future research within three main research domains: digital operational efficiency for banks, customer attraction strategies employed by digital banks, and the utilization of digital financial services in the retail industry.
Optimization of Nutrient-Rich Ice Plant (Mesembryanthemum crystallinum L.) Paste Fresh Noodle Pasta Using Response Surface Methodology
Authors: Kang Yeo-Wool , Joo Na-Mi
Published: June 2023

The ice plant is a species that is grown mainly in the dry regions of the American West and contains various minerals and ingredients beneficial for human health, such as inositol and beta-carotene. With the growing trend towards healthy foods, pasta consumption has also increased. Pasta is a convenient and low-glycemic-index food that is composed mainly of carbohydrates, proteins, lipids, dietary fiber, and trace amounts of minerals. The optimal mixing ratio was evaluated to produce pasta of the highest quality in terms of blood sugar elevation and antioxidant efficacy. The components and minerals of the ice plant, including D-pinitol and inositol, were analyzed, and 20 essential amino acids were identified. In this study, we also investigated the quality and characteristics of ice plant paste and eggs, as well as the quality, antioxidant activity, and formulation of raw materials mixed with ice plant at different ratios. Optimal conditions were found to be 46.73 g of ice plant paste in 100 g of durum wheat flour, 20.23 g of egg, and 2 g of salt, providing a way to develop fresh pasta that enhances the health benefits of ice plant paste without excessive moisture and other ingredients.
Discrete-Time Fractional Difference Calculus: Origins, Evolutions, and New Formalisms
Authors: Ortigueira Manuel Duarte
Published: June 2023

Differences are introduced as outputs of linear systems called differencers, being considered two classes: shift and scale-invariant. Several types are presented, namely: nabla and delta, bilateral, tempered, bilinear, stretching, and shrinking. Both continuous and discrete-time differences are described. ARMA-type systems based on differencers are introduced and exemplified. In passing, the incorrectness of the usual delta difference is shown.
Phylogenetic Analysis of Two New Mitochondrial Genomes of Singapora shinshana and Seriana bacilla from the Karst Region of Southwest China
Authors: Zhang Ni , Pu Tianyi , Wang Jinqiu , Tan Weiwen , Yuan Zhouwei , Li Can , Song Yuehua
Published: June 2023

Leafhoppers have been identified as a serious threat to different plants. To explore the characteristics of mitogenomes and reveal the phylogenetic positions of two species in the Typhlocybinae, complete mitogenomes of Singapora shinshana and Seriana bacilla were sequenced and annotated for the first time with lengths of 15,402 bp and 15,383 bp, respectively. The two mitogenomes contained 13 PCGs, 22 tRNA genes and 2 rRNA genes. The genome content, gene order, nucleotide composition, codon usage and amino acid composition are similar to those of other typical mitogenomes of Typhlocybinae. All 13 PCGs started with ATN codons, except for atp8 (TTA) and nad5 (TTG). All tRNAs were folded into a typical cloverleaf secondary structure, except for tRNA-Ser1 and tRNA-Val. Moreover, phylogenetic trees were constructed and analyzed based on all the PCGs from 42 mitogenomes using maximum likelihood (ML) and Bayesian inference (BI) methods. The results supported that eleven subfamilies are all monophyletic groups, S. shinshana and S. bacilla are members of Erythroneurini, but S. shinshana and the genus Empoascanara have a very close relationship with ((((Empoascanara sipra+ Empoascanara wengangensis) + Empoascanara dwalata) + Empoascanara gracilis) + S. shinshana), and S. bacilla is closely related to the genus Mitjaevia ((Mitjaevia dworakowskae + Mitjaevia shibingensis) + S. bacilla). These results provide valuable information for future study of evolutionary relationships in Typhlocybinae.
Revisiting the Role of Autophagy in Cardiac Differentiation: A Comprehensive Review of Interplay with Other Signaling Pathways
Authors: Kolahdouzmohammadi Mina , Kolahdouz-Mohammadi Roya , Tabatabaei Seyed Abdolhossein , Franco Brunella , Totonchi Mehdi
Published: June 2023

Autophagy is a critical biological process in which cytoplasmic components are sequestered in autophagosomes and degraded in lysosomes. This highly conserved pathway controls intracellular recycling and is required for cellular homeostasis, as well as the correct functioning of a variety of cellular differentiation programs, including cardiomyocyte differentiation. By decreasing oxidative stress and promoting energy balance, autophagy is triggered during differentiation to carry out essential cellular remodeling, such as protein turnover and lysosomal degradation of organelles. When it comes to controlling cardiac differentiation, the crosstalk between autophagy and other signaling networks such as fibroblast growth factor (FGF), Wnt, Notch, and bone morphogenetic proteins (BMPs) is essential, yet the interaction between autophagy and epigenetic controls remains poorly understood. Numerous studies have shown that modulating autophagy and precisely regulating it can improve cardiac differentiation, which can serve as a viable strategy for generating mature cardiac cells. These findings suggest that autophagy should be studied further during cardiac differentiation. The purpose of this review article is not only to discuss the relationship between autophagy and other signaling pathways that are active during the differentiation of cardiomyocytes but also to highlight the importance of manipulating autophagy to produce fully mature cardiomyocytes, which is a tough challenge.
Single-Cell Analysis in the Omics Era: Technologies and Applications in Cancer
Authors: Massimino Michele , Martorana Federica , Stella Stefania , Vitale Silvia Rita , Tomarchio Cristina , Manzella Livia , Vigneri Paolo
Published: June 2023

Cancer molecular profiling obtained with conventional bulk sequencing describes average alterations obtained from the entire cellular population analyzed. In the era of precision medicine, this approach is unable to track tumor heterogeneity and cannot be exploited to unravel the biological processes behind clonal evolution. In the last few years, functional single-cell omics has improved our understanding of cancer heterogeneity. This approach requires isolation and identification of single cells starting from an entire population. A cell suspension obtained by tumor tissue dissociation or hematological material can be manipulated using different techniques to separate individual cells, employed for single-cell downstream analysis. Single-cell data can then be used to analyze cell–cell diversity, thus mapping evolving cancer biological processes. Despite its unquestionable advantages, single-cell analysis produces massive amounts of data with several potential biases, stemming from cell manipulation and pre-amplification steps. To overcome these limitations, several bioinformatic approaches have been developed and explored. In this work, we provide an overview of this entire process while discussing the most recent advances in the field of functional omics at single-cell resolution.
The “FEEDS (FEeding Eating Deglutition Skills)” over Time Study in Cardiofaciocutaneous Syndrome
Authors: Onesimo Roberta , Sforza Elisabetta , Giorgio Valentina , Viscogliosi Germana , Kuczynska Eliza Maria , Margiotta Gaia , Perri Lucrezia , Limongelli Domenico , Proli Francesco , De Rose Cristina , Rigante Donato , Cerchiari Antonella , Tartaglia Marco , Leoni Chiara , Zampino Giuseppe
Published: June 2023

Feeding, eating and deglutition difficulties are key concerns in patients with cardiofaciocutaneous syndrome (CFCS). This study intends to quantify the development of feeding skills from birth to adulthood in patients with CFCS. Twenty-seven patients (eight males; mean age: 16.7 ± 8.3 years; median age: 15 years, age range: 1.5–38 years) with molecularly confirmed clinical diagnosis of CFCS were prospectively recruited from the Rare Disease Unit, Paediatrics Department, Fondazione Policlinico Agostino Gemelli-IRCCS, Rome, Italy, over a one-year period. Pathogenic variants along with key information regarding oro-motor features were collected. Sialorrhea was quantified using the Drooling Quotient 5. Feeding abilities were screened using the Italian version of the Montreal Children’s Hospital Feeding Scale (I-MCH-FS). The oral sensory processing section of the Sensory Profile completed the assessment. Mild-to-profuse drooling was experienced by 25% of patients, and food taste selectivity was a constant during infancy (65%), with persistence even beyond adolescence. Nineteen percent of participants with long-term enteral feeding dependency had BRAF, KRAS and MAP2K1 mutations. These findings document that mealtime challenges in CFCS do not remain restricted only to the paediatric age, and that supportive care until adulthood plays a key role.
Citizens’ Perspective on Coastal Erosion in Greece
Authors: Karkani Anna , Saitis Giannis , Komi Apostolia , Evelpidou Niki
Published: June 2023

Coastal erosion is a major coastal hazard in Greece. This work aimed to assess the perceptions of citizens about coastal erosion. We developed a questionnaire containing 25 questions that was distributed online and filled out by 1636 respondents. A surprising find was that 33% of the respondents were not aware of the phenomenon of coastal erosion. On the other hand, among those respondents who are aware of coastal erosion, there is a basic understanding of the major factors promoting erosion and its impacts. Responses also highlighted a lack of information and awareness from the media and public authorities, while the vast majority considered that protecting the coast from erosion should be important or a priority. Our findings stress the need for awareness-raising activities about this important natural hazard.
Staff Members’ Experience of Italian Shelters for LGBTQIA+ Homeless and Runaway People: An Exploratory Study
Authors: Tubertini Elena , Carbone Agostino , Santinello Massimo
Published: June 2023

Background: Some LGBTQIA+ people, after coming out, experience marginalization and homelessness due to rejection and discrimination from their family and community. The increase in support requests led to the creation of LGBTQIA+ temporary shelter homes worldwide. This study aims to explore the functioning and effectiveness of shelters, analyzing the experiences of staff members in Italy. Methods: Focus groups were held with a total of 15 staff members (age range: 32–53) working in three shelters for LGBTQIA+ people. Data were analyzed qualitatively through the grounded theory methodology. Results: Data coding showed five final core categories: (1) user characteristics; (2) staff characteristics; (3) community relations; (4) activities carried out by services; (5) criteria for intervention assessment and staff satisfaction. Results revealed some criticalities in the effectiveness of these services, particularly the difficulty in achieving autonomy for users, a weakness attributable to the non-exhaustive training of staff members and the funding discontinuity. Conclusion: To improve the efficacy of shelters, this study emphasizes the necessity to (a) carry out an analysis of the vulnerability of the local LGBTQIA+ community, (b) establish a stable network with local services (NHS system), and (c) implement staff members’ psychological training.
The Role of Selenium Nanoparticles in the Treatment of Liver Pathologies of Various Natures
Authors: Goltyaev Michael V. , Varlamova Elena G.
Published: June 2023

The liver is the body’s largest gland, and regulates a wide variety of physiological processes. The work of the liver can be disrupted in a variety of pathologies, the number of which is several hundred. It is extremely important to monitor the health of the liver and develop approaches to combat liver diseases. In recent decades, nanomedicine has become increasingly popular in the treatment of various liver pathologies, in which nanosized biomaterials, which are inorganic, polymeric, liposomal, albumin, and other nanoparticles, play an important role. Given the need to develop environmentally safe, inexpensive, simple, and high-performance biomedical agents for theragnostic purposes and showing few side effects, special attention is being paid to nanoparticles based on the important trace element selenium (Se). It is known that the metabolism of the microelement Se occurs in the liver, and its deficiency leads to the development of several serious diseases in this organ. In addition, the liver is the depot for most selenoproteins, which can reduce oxidative stress, inhibit tumor growth, and prevent other liver damage. This review is devoted to the description of the results of recent years, revealing the important role of selenium nanoparticles in the therapy and diagnosis of several liver pathologies, depending on the dose and physicochemical properties. The possibilities of selenium nanoparticles in the treatment of liver diseases, disclosed in the review, will not only reveal the advantages of their hepatoprotective properties but also significantly supplement the data on the role of the trace element selenium in the regulation of these diseases.
Unveiling Angiotensin II and Losartan-Induced Gene Regulatory Networks Using Human Urine-Derived Podocytes
Authors: Thimm Chantelle , Erichsen Lars , Wruck Wasco , Adjaye James
Published: June 2023

Podocytes are highly specialized cells that play a pivotal role in the blood filtration process in the glomeruli of the kidney, and their dysfunction leads to renal diseases. For this reason, the study and application of this cell type is of great importance in the field of regenerative medicine. Hypertension is mainly regulated by the renin–angiotensin–aldosterone system (RAAS), with its main mediator being angiotensin II (ANG II). Elevated ANG II levels lead to a pro-fibrotic, inflammatory, and hypertrophic milieu that induces apoptosis in podocytes. The activation of RAAS is critical for the pathogenesis of podocyte injury; as such, to prevent podocyte damage, patients with hypertension are administered drugs that modulate RAAS signaling. A prime example is the orally active, non-peptide, selective angiotensin-II-type I receptor (AGTR1) blocker losartan. Here, we demonstrate that SIX2-positive urine-derived renal progenitor cells (UdRPCs) and their immortalized counterpart (UM51-hTERT) can be directly differentiated into mature podocytes. These podocytes show activation of RAAS after stimulation with ANG II, resulting in ANG II-dependent upregulation of the expression of the angiotensin-II-type I receptor, AGTR1, and the downregulated expression of the angiotensin-II-type II receptor 2 (AGTR2). The stimulation of podocytes with losartan counteracts ANG II-dependent changes, resulting in a dependent favoring of the specific receptor from AGTR1 to AGTR2. Transcriptome analysis revealed 94 losartan-induced genes associated with diverse biological processes and pathways such as vascular smooth muscle contraction, the oxytocin signaling pathway, renin secretion, and ECM-receptor interaction. Co-stimulation with losartan and ANG II induced the exclusive expression of 106 genes associated with DNA methylation or demethylation, cell differentiation, the developmental process, response to muscle stretch, and calcium ion transmembrane transport. These findings highlight the usefulness of UdRPC-derived podocytes in studying the RAAS pathway and nephrotoxicity in various kidney diseases.
Methylammonium Tetrel Halide Perovskite Ion Pairs and Their Dimers: The Interplay between the Hydrogen-, Pnictogen- and Tetrel-Bonding Interactions
Authors: Varadwaj Pradeep R. , Varadwaj Arpita , Marques Helder M. , Yamashita Koichi
Published: June 2023

The structural stability of the extensively studied organic–inorganic hybrid methylammonium tetrel halide perovskite semiconductors, MATtX3 (MA = CH3NH3+; Tt = Ge, Sn, Pb; X = Cl, Br, I), arises as a result of non-covalent interactions between an organic cation (CH3NH3+) and an inorganic anion (TtX3−). However, the basic understanding of the underlying chemical bonding interactions in these systems that link the ionic moieties together in complex configurations is still limited. In this study, ion pair models constituting the organic and inorganic ions were regarded as the repeating units of periodic crystal systems and density functional theory simulations were performed to elucidate the nature of the non-covalent interactions between them. It is demonstrated that not only the charge-assisted N–H···X and C–H···X hydrogen bonds but also the C–N···X pnictogen bonds interact to stabilize the ion pairs and to define their geometries in the gas phase. Similar interactions are also responsible for the formation of crystalline MATtX3 in the low-temperature phase, some of which have been delineated in previous studies. In contrast, the Tt···X tetrel bonding interactions, which are hidden as coordinate bonds in the crystals, play a vital role in holding the inorganic anionic moieties (TtX3−) together. We have demonstrated that each Tt in each [CH3NH3+•TtX3−] ion pair has the capacity to donate three tetrel (σ-hole) bonds to the halides of three nearest neighbor TtX3− units, thus causing the emergence of an infinite array of 3D TtX64− octahedra in the crystalline phase. The TtX44− octahedra are corner-shared to form cage-like inorganic frameworks that host the organic cation, leading to the formation of functional tetrel halide perovskite materials that have outstanding optoelectronic properties in the solid state. We harnessed the results using the quantum theory of atoms in molecules, natural bond orbital, molecular electrostatic surface potential and independent gradient models to validate these conclusions.
Expression-Based Diagnosis, Treatment Selection, and Drug Development for Breast Cancer
Authors: Ye Qing , Wang Jiajia , Ducatman Barbara , Raese Rebecca A. , Rogers Jillian L. , Wan Ying-Wooi , Dong Chunlin , Padden Lindsay , Pugacheva Elena N. , Qian Yong , Guo Nancy Lan
Published: June 2023

There is currently no gene expression assay that can assess if premalignant lesions will develop into invasive breast cancer. This study sought to identify biomarkers for selecting patients with a high potential for developing invasive carcinoma in the breast with normal histology, benign lesions, or premalignant lesions. A set of 26-gene mRNA expression profiles were used to identify invasive ductal carcinomas from histologically normal tissue and benign lesions and to select those with a higher potential for future cancer development (ADHC) in the breast associated with atypical ductal hyperplasia (ADH). The expression-defined model achieved an overall accuracy of 94.05% (AUC = 0.96) in classifying invasive ductal carcinomas from histologically normal tissue and benign lesions (n = 185). This gene signature classified cancer development in ADH tissues with an overall accuracy of 100% (n = 8). The mRNA expression patterns of these 26 genes were validated using RT-PCR analyses of independent tissue samples (n = 77) and blood samples (n = 48). The protein expression of PBX2 and RAD52 assessed with immunohistochemistry were prognostic of breast cancer survival outcomes. This signature provided significant prognostic stratification in The Cancer Genome Atlas breast cancer patients (n = 1100), as well as basal-like and luminal A subtypes, and was associated with distinct immune infiltration and activities. The mRNA and protein expression of the 26 genes was associated with sensitivity or resistance to 18 NCCN-recommended drugs for treating breast cancer. Eleven genes had significant proliferative potential in CRISPR-Cas9/RNAi screening. Based on this gene expression signature, the VEGFR inhibitor ZM-306416 was discovered as a new drug for treating breast cancer.
Comparative Transcriptome Profiling Unfolds a Complex Defense and Secondary Metabolite Networks Imparting Corynespora cassiicola Resistance in Soybean (Glycine max (L.) Merrill)
Authors: Patel Sejal , Patel Jinesh , Silliman Katherine , Hall Nathan , Bowen Kira , Koebernick Jenny
Published: June 2023

Target spot is caused by Corynespora cassiicola, which heavily affects soybean production areas that are hot and humid. Resistant soybean genotypes have been identified; however, the molecular mechanisms governing resistance to infection are unknown. Comparative transcriptomic profiling using two known resistant genotypes and two susceptible genotypes was performed under infected and control conditions to understand the regulatory network operating between soybean and C. cassiicola. RNA-Seq analysis identified a total of 2571 differentially expressed genes (DEGs) which were shared by all four genotypes. These DEGs are related to secondary metabolites, immune response, defense response, phenylpropanoid, and flavonoid/isoflavonoid pathways in all four genotypes after C. cassiicola infection. In the two resistant genotypes, additional upregulated DEGs were identified affiliated with the defense network: flavonoids, jasmonic acid, salicylic acid, and brassinosteroids. Further analysis led to the identification of differentially expressed transcription factors, immune receptors, and defense genes with a leucine-rich repeat domain, dirigent proteins, and cysteine (C)-rich receptor-like kinases. These results will provide insight into molecular mechanisms of soybean resistance to C. cassiicola infection and valuable resources to potentially pyramid quantitative resistance loci for improving soybean germplasm.
Oyster Mushroom Spherical Virus Crosses the Species Barrier and Is Pathogenic to a New Host Pleurotus pulmonarius
Authors: Zhang Xiaoyan , Hu Haijing , Zhao Yanxiang , Wang Yifan , Zhang Wenjing , You Lunhe , Wang Jianrui , Liu Yu , Cheng Xianhao
Published: June 2023

Oyster mushroom spherical virus (OMSV) is a mycovirus with a positive-sense single-stranded RNA genome that infects the edible mushroom Pleurotus ostreatus. OMSV is horizontally transferred from an infected strain to a cured strain via mycelia. The infection results in significant inhibition of mycelial growth, malformation of fruiting bodies, and yield loss in oyster mushrooms. This study successfully transferred OMSV from P. ostreatus to Pleurotus pulmonarius. However, transmission was not successful in other Pleurotus species including P. citrinopileatus, P. eryngii, P. nebrodensis, and P. salmoneostramineus. The successful OMSV infection in P. pulmonarius was further verified with Western blot analysis using a newly prepared polyclonal antiserum against the OMSV coat protein. Furthermore, OMSV infection reduced the mycelial growth rate of P. pulmonarius. The OMSV-infected strain demonstrated abnormal performance including twisted mushrooms or irregular edge of the cap as well as reduced yield of fruiting bodies in P. pulmonarius, compared to the OMSV-free strain. This study is the first report on the infection and pathogenicity of OMSV to the new host P. pulmonarius. The data from this study therefore suggest that OMSV is a potential threat to P. pulmonarius.
Double-Labeling Method for Visualization and Quantification of Membrane-Associated Proteins in Lactococcus lactis
Authors: Hoang Mai Ngoc , Peterbauer Clemens
Published: June 2023

Lactococcus lactis displaying recombinant proteins on its surface can be used as a potential drug delivery vector in prophylactic medication and therapeutic treatments for many diseases. These applications enable live-cell mucosal and oral administration, providing painless, needle-free solutions and triggering robust immune response at the site of pathogen entry. Immunization requires quantitative control of antigens and, ideally, a complete understanding of the bacterial processing mechanism applied to the target proteins. In this study, we propose a double-labeling method based on a conjugated dye specific for a recombinantly introduced polyhistidine tag (to visualize surface-exposed proteins) and a membrane-permeable dye specific for a tetra-cysteine tag (to visualize cytoplasmic proteins), combined with a method to block the labeling of surface-exposed tetra-cysteine tags, to clearly obtain location-specific signals of the two dyes. This allows simultaneous detection and quantification of targeted proteins on the cell surface and in the cytoplasm. Using this method, we were able to detect full-length peptide chains for the model proteins HtrA and BmpA in L. lactis, which are associated with the cell membrane by two different attachment modes, and thus confirm that membrane-associated proteins in L. lactis are secreted using the Sec-dependent post-translational pathway. We were able to quantitatively follow cytoplasmic protein production and accumulation and subsequent export and surface attachment, which provides a convenient tool for monitoring these processes for cell surface display applications.
Green Synthesis and the Evaluation of a Functional Amphiphilic Block Copolymer as a Micellar Curcumin Delivery System
Authors: Kalinova Radostina , Grancharov Georgy , Doumanov Jordan , Mladenova Kirilka , Petrova Svetla , Dimitrov Ivaylo
Published: June 2023

Polymer micelles represent one of the most attractive drug delivery systems due to their design flexibility based on a variety of macromolecular synthetic methods. The environmentally safe chemistry in which the use or generation of hazardous materials is minimized has an increasing impact on polymer-based drug delivery nanosystems. In this work, a solvent-free green synthetic procedure was applied for the preparation of an amphiphilic diblock copolymer consisting of biodegradable hydrophobic poly(acetylene-functional carbonate) and biocompatible hydrophilic polyethylene glycol (PEG) blocks. The cyclic functional carbonate monomer 5-methyl-5-propargyloxycarbonyl-1,3-dioxane-2-one (MPC) was polymerized in bulk using methoxy PEG-5K as a macroinitiator by applying the metal-free organocatalyzed controlled ring-opening polymerization at a relatively low temperature of 60 °C. The functional amphiphilic block copolymer self-associated in aqueous media into stable micelles with an average diameter of 44 nm. The copolymer micelles were physico-chemically characterized and loaded with the plant-derived anticancer drug curcumin. Preliminary in vitro evaluations indicate that the functional copolymer micelles are non-toxic and promising candidates for further investigation as nanocarriers for biomedical applications.
Protein Extract of a Probiotic Strain of Hafnia alvei and Bacterial ClpB Protein Improve Glucose Tolerance in Mice
Authors: Zolotarev Vasiliy A. , Murovets Vladimir O. , Sepp Anastasiya L. , Sozontov Egor A. , Lukina Ekaterina A. , Khropycheva Raisa P. , Pestereva Nina S. , Ivleva Irina S. , El Mehdi Mouna , Lahaye Emilie , Chartrel Nicolas , Fetissov Sergueï O.
Published: June 2023

A commercial strain of Hafnia alvei (H. alvei) 4597 bacteria was shown to reduce food intake and promote weight loss, effects possibly induced by the bacterial protein ClpB, an antigen-mimetic of the anorexigenic α-melanocyte-stimulating hormone. A decrease in the basal plasma glucose levels was also observed in overweight fasted humans and mice receiving H. alvei. However, it is not known whether H. alvei influences sweet taste preference and whether its protein extract or ClpB are sufficient to increase glucose tolerance; these are the objectives tested in the present study. C57BL/6J male mice were kept under standard diet and were gavaged daily for 17 days with a suspension of H. alvei (4.5 × 107 CFU/animal) or with H. alvei total protein extract (5 μg/animal) or saline as a control. Sweet taste preference was analyzed via a brief-access licking test with sucrose solution. Glucose tolerance tests (GTT) were performed after the intraperitoneal (IP) or intragastric (IG) glucose administration at the 9th and 15th days of gavage, respectively. The expression of regulatory peptides’ mRNA levels was assayed in the hypothalamus. In another experiment performed in non-treated C57BL/6J male mice, effects of acute IP administration of recombinant ClpB protein on glucose tolerance were studied by both IP- and IG-GTT. Mice treated with the H. alvei protein extract showed an improved glucose tolerance in IP-GTT but not in IG-GTT. Both groups treated with H. alvei bacteria or protein extract showed a reduction of pancreatic tissue weight but without significant changes to basal plasma insulin. No significant effects of H. alvei bacteria or its total protein extract administration were observed on the sweet taste preference, insulin tolerance and expression of regulatory peptides’ mRNA in the hypothalamus. Acute administration of ClpB in non-treated mice increased glucose tolerance during the IP-GTT but not the IG-GTT, and reduced basal plasma glucose levels. We conclude that both the H. alvei protein extract introduced orally and the ClpB protein administered via IP improve glucose tolerance probably by acting at the glucose postabsorptive level. Moreover, H. alvei probiotic does not seem to influence the sweet taste preference. These results justify future testing of both the H. alvei protein extract and ClpB protein in animal models of diabetes.
Repetitive Low-Level Blast Exposure via Akt/NF-κB Signaling Pathway Mediates the M1 Polarization of Mouse Alveolar Macrophage MH-S Cells
Authors: Geng Chenhao , Wang Xinyue , Chen Jiale , Sun Na , Wang Yuru , Li Zizheng , Han Lu , Hou Shike , Fan Haojun , Li Ning , Gong Yanhua
Published: June 2023

Repetitive low-level blast (rLLB) exposure is a potential risk factor for the health of soldiers or workers who are exposed to it as an occupational characteristic. Alveolar macrophages (AMs) are susceptible to external blast waves and produce pro-inflammatory or anti-inflammatory effects. However, the effect of rLLB exposure on AMs is still unclear. Here, we generated rLLB waves through a miniature manual Reddy-tube and explored their effects on MH-S cell morphology, phenotype transformation, oxidative stress status, and apoptosis by immunofluorescence, real-time quantitative PCR (qPCR), western blotting (WB) and flow cytometry. Ipatasertib (GDC-0068) or PDTC was used to verify the role of the Akt/NF-κB signaling pathway in these processes. Results showed that rLLB treatment could cause morphological irregularities and cytoskeletal disorders in MH-S cells and promote their polarization to the M1 phenotype by increasing iNOS, CD86 and IL-6 expression. The molecular mechanism is through the Akt/NF-κB signaling pathway. Moreover, we found reactive oxygen species (ROS) burst, Ca2+ accumulation, mitochondrial membrane potential reduction, and early apoptosis of MH-S cells. Taken together, our findings suggest rLLB exposure may cause M1 polarization and early apoptosis of AMs. Fortunately, it is blocked by specific inhibitors GDC-0068 or PDTC. This study provides a new treatment strategy for preventing and alleviating health damage in the occupational population caused by rLLB exposure.
Ion-Powered Rotary Motors: Where Did They Come from and Where They Are Going?
Authors: Nandel Vibhuti , Scadden Jacob , Baker Matthew A. B.
Published: June 2023

Molecular motors are found in many living organisms. One such molecular machine, the ion-powered rotary motor (IRM), requires the movement of ions across a membrane against a concentration gradient to drive rotational movement. The bacterial flagellar motor (BFM) is an example of an IRM which relies on ion movement through the stator proteins to generate the rotation of the flagella. There are many ions which can be used by the BFM stators to power motility and different ions can be used by a single bacterium expressing multiple stator variants. The use of ancestral sequence reconstruction (ASR) and functional analysis of reconstructed stators shows promise for understanding how these proteins evolved and when the divergence in ion use may have occurred. In this review, we discuss extant BFM stators and the ions that power them as well as recent examples of the use of ASR to study ion-channel selectivity and how this might be applied to further study of the BFM stator complex.
Microbial Metabolites of 3-n-butylphthalide as Monoamine Oxidase A Inhibitors
Authors: Gach Joanna , Grzelczyk Joanna , Strzała Tomasz , Boratyński Filip , Olejniczak Teresa
Published: June 2023

Novel compounds with antidepressant activity via monoamine oxidase inhibition are being sought. Among these, derivatives of 3-n-butylphthalide, a neuroprotective lactone from Apiaceae plants, may be prominent candidates. This study aimed to obtain the oxidation products of 3-n-butylphthalide and screen them regarding their activity against the monoamine oxidase A (MAO-A) isoform. Such activity of these compounds has not been previously tested. To obtain the metabolites, we used fungi as biocatalysts because of their high oxidative capacity. Overall, 37 strains were used, among which Penicillium and Botrytis spp. were the most efficient, leading to the obtaining of three main products: 3-n-butyl-10-hydroxyphthalide, 3-n-butylphthalide-11-oic acid, and 3-n-butyl-11-hydroxyphthalide, with a total yield of 0.38–0.82 g per g of the substrate, depending on the biocatalyst used. The precursor–3-n-butylphthalide and abovementioned metabolites inhibited the MAO-A enzyme; the most active was the carboxylic acid derivative of the lactone with inhibitory constant (Ki) < 0.001 µmol/L. The in silico prediction of the drug-likeness of the metabolites matches the assumptions of Lipinski, Ghose, Veber, Egan, and Muegge. All the compounds are within the optimal range for the lipophilicity value, which is connected to adequate permeability and solubility.
Genetic Ablation of Inositol 1,4,5-Trisphosphate Receptor Type 2 (IP3R2) Fails to Modify Disease Progression in a Mouse Model of Spinocerebellar Ataxia Type 3
Authors: Cunha-Garcia Daniela , Monteiro-Fernandes Daniela , Correia Joana Sofia , Neves-Carvalho Andreia , Vilaça-Ferreira Ana Catarina , Guerra-Gomes Sónia , Viana João Filipe , Oliveira João Filipe , Teixeira-Castro Andreia , Maciel Patrícia , Duarte-Silva Sara
Published: June 2023

Spinocerebellar ataxia type 3 (SCA3) is a rare neurodegenerative disease caused by an abnormal polyglutamine expansion within the ataxin-3 protein (ATXN3). This leads to neurodegeneration of specific brain and spinal cord regions, resulting in a progressive loss of motor function. Despite neuronal death, non-neuronal cells, including astrocytes, are also involved in SCA3 pathogenesis. Astrogliosis is a common pathological feature in SCA3 patients and animal models of the disease. However, the contribution of astrocytes to SCA3 is not clearly defined. Inositol 1,4,5-trisphosphate receptor type 2 (IP3R2) is the predominant IP3R in mediating astrocyte somatic calcium signals, and genetically ablation of IP3R2 has been widely used to study astrocyte function. Here, we aimed to investigate the relevance of IP3R2 in the onset and progression of SCA3. For this, we tested whether IP3R2 depletion and the consecutive suppression of global astrocytic calcium signalling would lead to marked changes in the behavioral phenotype of a SCA3 mouse model, the CMVMJD135 transgenic line. This was achieved by crossing IP3R2 null mice with the CMVMJD135 mouse model and performing a longitudinal behavioral characterization of these mice using well-established motor-related function tests. Our results demonstrate that IP3R2 deletion in astrocytes does not modify SCA3 progression.
Association of Low Serum Bilirubin Concentrations and Promoter Variations in the UGT1A1 and HMOX1 Genes with Type 2 Diabetes Mellitus in the Czech Population
Authors: Jirásková Alena , Škrha Jan , Vítek Libor
Published: June 2023

Bilirubin has potent biological beneficial effects, protecting against atherosclerosis, obesity, and metabolic syndrome. The aim of this study was to assess serum bilirubin concentrations and (TA)n and (GT)n microsatellite variations in the promoter regions of the UGT1A1 and HMOX1 genes, respectively, in patients with type 2 diabetes mellitus (T2DM). The study was carried out in 220 patients with T2DM and 231 healthy control subjects, in whom standard biochemical tests were performed. The (TA)n and (GT)n dinucleotide variations were determined by means of fragment (size-based) analysis using an automated capillary DNA sequencer. Compared to controls, both male and female patients with T2DM had lower serum bilirubin concentrations (9.9 vs. 12.9 μmol/L, and 9.0 vs. 10.6 μmol/L, in men and women, respectively, p < 0.001). Phenotypic Gilbert syndrome was much less prevalent in T2DM patients, as was the frequency of the (TA)7/7UGT1A1 genotype in male T2DM patients. (GT)nHMOX1 genetic variations did not differ between diabetic patients and controls. Our results demonstrate that the manifestation of T2DM is associated with lower serum bilirubin concentrations. Consumption of bilirubin due to increased oxidative stress associated with T2DM seems to be the main explanation, although (TA)n repeat variations in UGT1A1 partially contribute to this phenomenon.
Quantitative Proteomic Analysis on the Slightly Acidic Electrolyzed Water Triggered Viable but Non-Culturable Listeria monocytogenes
Authors: Chang Hsin-Yi , Gui Chin-Ying , Huang Tsui-Chin , Hung Yen-Con , Chen Tai-Yuan
Published: June 2023

This study undertakes a comprehensive exploration of the impact of slightly acidic electrolyzed water (SAEW) on Listeria monocytogenes, a common foodborne pathogen, with a particular focus on understanding the molecular mechanisms leading to the viable but nonculturable (VBNC) state. Given the widespread application of SAEW as an effective disinfectant in the food industry, uncovering these molecular pathways is crucial for improving food safety measures. We employed tandem mass tags (TMT), labeling proteomic techniques and LC-MS/MS to identify differentially expressed proteins under two doses of SAEW conditions. We indicated 203 differential expressed proteins (DEPs), including 78 up-regulated and 125 down-regulated DEPs. The functional enrichment analysis of these proteins indicated that ribosomes, biosynthesis of secondary metabolites, and aminoacyl-tRNA biosynthesis were enriched functions affected by SAEW. Further, we delved into the role of protein chlorination, a potential consequence of reactive chlorine species generated during the SAEW production process, by identifying 31 chlorinated peptides from 22 proteins, with a dominant sequence motif of Rxxxxx[cY] and functionally enriched in translation. Our findings suggest that SAEW might prompt alterations in the protein translation process and trigger compensatory ribosome biosynthesis. However, an imbalance in the levels of elongation factors and AARSs could hinder recovery, leading to the VBNC state. This research carries substantial implications for food safety and sanitation, as it adds to our understanding of the SAEW-induced VBNC state in L. monocytogenes and offers potential strategies for its control.
Exosome–Autophagy Crosstalk in Enveloped Virus Infection
Authors: Wang Yuqi , Ren Linzhu , Bai Haocheng , Jin Qing , Zhang Liying
Published: June 2023

Exosomes, which are extracellular vesicles (EVs) predominantly present in bodily fluids, participate in various physiological processes. Autophagy, an intracellular degradation mechanism, eliminates proteins and damaged organelles by forming double-membrane autophagosomes. These autophagosomes subsequently merge with lysosomes for target degradation. The interaction between autophagy and endosomal/exosomal pathways can occur at different stages, exerting significant influences on normal physiology and human diseases. The interplay between exosomes and the autophagy pathway is intricate. Exosomes exhibit a cytoprotective role by inducing intracellular autophagy, while autophagy modulates the biogenesis and degradation of exosomes. Research indicates that exosomes and autophagy contribute to the infection process of numerous enveloped viruses. Enveloped viruses, comprising viral nucleic acid, proteins, or virions, can be encapsulated within exosomes and transferred between cells via exosomal transport. Consequently, exosomes play a crucial role in the infection of certain viral diseases. This review presents recent findings on the interplay between exosomes and autophagy, as well as their implications in the infection of enveloped viruses, thereby offering valuable insights into the pathogenesis and vaccine research of enveloped virus infection.
Chronic Liver Disease: Latest Research in Pathogenesis, Detection and Treatment
Authors: De Siervi Silvia , Cannito Stefania , Turato Cristian
Published: June 2023

Chronic liver disease (CLD) is a major global health threat and has emerged as a leading cause of human death [...]
Fads2b Plays a Dominant Role in ∆6/∆5 Desaturation Activities Compared with Fads2a in Common Carp (Cyprinus carpio)
Authors: Zhao Ran , Yang Chen-Ru , Wang Ya-Xin , Xu Zi-Ming , Li Shang-Qi , Li Jin-Cheng , Sun Xiao-Qing , Wang Hong-Wei , Wang Qi , Zhang Yan , Li Jiong-Tang
Published: June 2023

Highly unsaturated fatty acids (HUFAs) are essential for mammalian health, development and growth. However, most mammals, including humans, are incapable of synthesizing n-6 and n-3 HUFAs. Fish can convert C18 unsaturated fatty acids into n-6 and n-3 HUFAs via fatty acid desaturase (Fads), in which Fads2 is a key enzyme in HUFA biosynthesis. The allo-tetraploid common carp theoretically encode two duplicated fads2 genes. The expression patterns and desaturase functions of these two homologous genes are still unknown. In this study, the full length of the fads2a and fads2b were identified in common carp (Cyprinus carpio). Expression analyses indicate that both genes were mainly expressed in the liver and the expression of fads2b is higher than fads2a at different developmental stages in carp embryos. Heterogenous expression and 3D docking analyses suggested that Fads2b demonstrated stronger ∆6 and ∆5 desaturase activities than Fads2a. The core promotor regions of fads2a and fads2b were characterized and found to have different potential transcriptional binding sites. These results revealed the same desaturase functions, but different activities of two homologues of fasd2 genes in common carp. The data showed that fads2b played a more important role in HUFA synthesis through both expression and functional analyses.
Small RNA Profiling in an HTLV-1-Infected Patient with Acute Adult T-Cell Leukemia-Lymphoma at Diagnosis and after Maintenance Therapy: A Case Study
Authors: Pessôa Rodrigo , de Souza Daniela Raguer Valadão , Nukui Youko , Pereira Juliana , Fernandes Lorena Abreu , Marcusso Rosa Nascimento , de Oliveira Augusto César Penalva , Casseb Jorge , da Silva Duarte Alberto José , Sanabani Sabri Saeed
Published: June 2023

Small RNAs (sRNAs) are epigenetic regulators of essential biological processes associated with the development and progression of leukemias, including adult T-cell leukemia/lymphoma (ATLL) caused by human T-cell lymphotropic virus type 1 (HTLV-1), an oncogenic human retrovirus originally discovered in a patient with adult T-cell leukemia/lymphoma. Here, we describe the sRNA profile of a 30-year-old woman with ATLL at the time of diagnosis and after maintenance therapy with the aim of correlating expression levels with response to therapy.