bluuebunny/crossref_metadata_2025 huggingface dataset:
The Impact of Expenditure on Research and Development on Selected Energy Factors in the European Union
Authors: Chmielewski Wojciech , Postuła Marta , Dubel Przemysław
Published: April 2023

Due to the fact that the European Union is striving to achieve its sustainable development goals, in particular goal No. 7, which is to provide users with low-emission, and cheap access to, energy, this article’s aim is to verify whether there is a relationship between R&D expenditure and key energy variables in the EU countries in 2010–2020. Data on R&D expenditures incurred by the EU Member States in the period 2010–2020 were used for the research and were tested using the Autoregressive Distributed Lags Model (ARDL). The study identified a strong positive relationship between total R&D expenditure and the increase in energy consumption from renewable energy sources, and a smaller impact of total R&D expenditure as well as enterprise R&D expenditure on the increase in fossil energy consumption. Also noted was a weak relationship between R&D expenditure and electricity and gas prices for both household and non-household customers. The obtained results prove that in the context of the level of implementation of SDG No. 7 in the EU countries, R&D expenditure results in greater access to low-emission energy from renewable sources, but the achievement of the aforementioned sustainable development goal in other aspects (reduction in energy consumption from fossil fuels and ensuring lower price energy) is not possible with the current level of R&D expenditure.
Endocrine-Disrupting Chemicals and Persistent Organic Pollutants in Infant Formulas and Baby Food: Legislation and Risk Assessments
Authors: Hatzidaki Eleftheria , Pagkalou Marina , Katsikantami Ioanna , Vakonaki Elena , Kavvalakis Matthaios , Tsatsakis Aristidis M. , Tzatzarakis Manolis N.
Published: April 2023

Human milk is the healthiest option for newborns, although, under specific circumstances, infant formula is a precious alternative for feeding the baby. Except for the nutritional content, infant formulas and baby food must be pollutant-free. Thus, their composition is controlled by continuous monitoring and regulated by establishing upper limits and guideline values for safe exposure. Legislation differs worldwide, although there are standard policies and strategies for protecting vulnerable infants. This work presents current regulations and directives for restricting endocrine-disrupting chemicals and persistent organic pollutants in infant formulas. Risk assessment studies, which are limited, are necessary to depict exposure variations and assess the health risks for infants from dietary exposure to pollutants.
COPD and Depression Analysis in Regard to Obstructive Pulmonary Levels
Authors: Jelić Ivana , Mihajlović Goran , Mihajlović Filip , Minić Nataša , Ratinac Miloš , Pantović-Stefanović Maja
Published: April 2023

Depression symptoms take place recurrently in patients suffering from COPD. This study aims to assess the effects of antidepressant therapy in patients with COPD and a depressive disorder in relation to COPD levels. The study population consisted of N = 87 patients diagnosed with COPD, according to the GOLD criteria, and a depressive disorder. All of the patients were subjected to clinical and psychiatric exploration according to psychiatric assessment instruments, which was followed by SSRI therapy for the duration of 8 weeks. The main methods used were descriptive statistics and analysis of variance. The results showed a different distribution of depressive symptoms at a different stage of COPD by FEV1 (χ2 = 30.47, df = 6, p < 0.01) and by mMRC (χ2 = 34.6, df = 6, p < 0.01). After the application of SSRIs, there was a significant improvement in HDRS scores in all stages of COPD by FEV1 (χ2 = 251.62, df = 9, p < 0.01) and by mMRC (χ2 = 919.17, df = 9, p < 0.01). This study contributes to the improvement in the quality of life of patients by the targeted application of SSRI therapy and, therefore, more precise and better overall treatment results.
Addressing Tobacco Use in Underserved Communities Outside of Primary Care: The Need to Tailor Tobacco Cessation Training for Community Health Workers
Authors: Tan Marcia M. , Oke Shariwa , Ellison Daryn , Huard Clarissa , Veluz-Wilkins Anna
Published: April 2023

Individuals from communities with a low socioeconomic status have the highest rates of tobacco use but are less likely to receive assistance with quitting. Community health workers (CHWs) are well-positioned to engage these communities; however, CHWs face barriers in receiving relevant tobacco cessation training. The objective of this study was to conduct a mixed methods needs assessment to describe tobacco practices and the desire for training among CHWs. After incorporating CHW feedback, we developed a needs assessment survey to understand knowledge, practices, and attitudes about tobacco cessation in Chicago, IL. CHWs (N = 23) recruited from local community-based organizations completed the survey online or in-person. We then conducted a focus group with CHWs (N = 6) to expand upon the survey and used the Framework Method to analyze the qualitative data. CHWs reported that their clients had low incomes, low literacy levels, and high smoking rates (e.g., “99%” of patients). About 73.3% reported discussing tobacco use during visits, but fewer reported that they had provided cessation advice (43%) or intervened directly (9%). CHWs described high variability in their work environments (e.g., location, duration, content of visits, etc.) and greater continuity of care. CHWs discussed that existing training on how to conduct tobacco interventions is ineffective, because of its stand-alone design. Our findings illustrate how CHWs adapt to their clients’ needs, and that the currently available “gold-standard” cessation curricula are incompatible with the training needs and flexible care delivery model of CHWs. A curriculum tailored to the CHW experience is needed to maximize the strengths of the CHW care model by training CHWs to adaptively intervene regarding tobacco use in their highly burdened patients.
What Do We Know about Suicide Bereavement, and What We Can Do to Help Suicide-Loss Survivors?
Authors: Levi-Belz Yossi , Krysinska Karolina , Andriessen Karl
Published: April 2023

“Suicide is not only the end of life for the deceased but also the beginning of a highly challenging life for those left behind” [...]
Changes in Physical Performance among Community-Dwelling Older Adults in Six Years
Authors: Gomes Darlise Rodrigues dos Passos , Santos Leonardo Pozza , Gonzalez Maria Cristina , Vieira Edgar Ramos , Bielemann Renata Moraes
Published: April 2023

Changes in physical performance (PP) occur with aging, and understanding the magnitude of these changes over time is important. This study evaluated changes in Gait Speed (GS) and Timed Up and Go (TUG) performance and their association with related factors among community-dwelling older adults over a period of five to six years. A cohort study of 476 older adults with baseline assessment conducted in 2014 and reassessment in 2019–2020 was carried out. Associations between PP changes over time and sociodemographic, behavioral, and health variables were evaluated using mixed linear models. Approximately 68% of the participants declined PP; 20% had no relevant change in GS and 9% in TUG time (sustained PP); 12% increased GS, and 23% decreased TUG time (improved PP). Being male (p = 0.023), living without a partner/separated (p = 0.035), higher education (p = 0.019), and alcohol consumption in the prior month (p = 0.045) were associated with decreased GS, while older age (p < 0.001), having lower socioeconomic status (p < 0.004), physical inactivity (p = 0.017), and being overweight (p = 0.007) were associated with increased TUG time. PP declined for most participants. Factors most strongly associated with PP decline are non-modifiable. The high prevalence of PP decline over time signals the importance of including physical tests in yearly health assessments.
Sustaining and Expanding Internet-Delivered Cognitive Behavioral Therapy (ICBT) for Public Safety Personnel across Canada: A Survey of Stakeholder Perspectives
Authors: Landry Caeleigh A. , Beahm Janine D. , McCall Hugh C. , Hadjistavropoulos Heather D.
Published: April 2023

Public safety personnel (PSP) experience an elevated risk of mental health problems and face barriers to treatment. Internet-delivered cognitive behavioral therapy (ICBT) has been tailored to PSP to improve access to mental health care. In this study, we sought to investigate perceptions of ICBT, particularly among those with and without prior knowledge of ICBT and between PSP leaders and non-leaders. A survey was administered to 524 PSP from across Canada to identify (a) how PSP perceive ICBT, (b) the extent of organizational support for tailored ICBT in PSP organizations, particularly leadership’s support, and (c) perceived facilitators and barriers to funding tailored ICBT. The results indicated that PSP perceive ICBT to have more advantages than disadvantages. PSP who had previously heard of tailored ICBT had more positive perceptions. PSP indicated that there is a need for ICBT, and PSP leaders indicated their support for the implementation of tailored ICBT. The study identified that there is a need for increasing awareness of the effectiveness of and need for ICBT in order to facilitate funding of services. Overall, the current study indicates that PSP support ICBT as a valued form of therapy and that policy makers and service providers seeking to provide ICBT to PSP may increase support for ICBT services through more education and awareness.
Knowledge of Obstetric Danger Signs among Pregnant Women in the Eastern Democratic Republic of the Congo
Authors: Imani Ramazani Bin-Eradi , Mabakutuvangilanga Ntala Simon-Decap , Katuashi Ishoso Daniel , Rothan-Tondeur Monique
Published: April 2023

A lack of awareness regarding obstetric danger signs (ODS) is one of the factors that delay a pregnant woman’s decision to seek emergency obstetric care. In developing countries, this delay can lead to high morbidity and mortality among pregnant women. In eastern Democratic Republic of Congo (DRC), very few studies have been conducted to assess the level of knowledge of pregnant women about ODS. Therefore, this study aimed to assess the knowledge of pregnant women about ODS in health facilities in eastern DRC. This quantitative cross-sectional, descriptive, and analytical study was conducted in 19 health facilities in the Kasongo health zone in the south Maniema Province of eastern DRC. A total of 624 pregnant women aged 12–49 years were interviewed in this study. Of these, 60.6% were secondary school graduates, >99% were married, 85.5% were cultivators, and 67.9% were Muslims. The knowledge of ODS among pregnant women was low (21.9%). The most cited danger signs during pregnancy, labor/delivery, and postpartum included severe abdominal pain and severe vaginal bleeding. Additionally, pregnant women aged 30–39 years (p = 0.015) and those who had given birth once (p = 0.049), twice (p = 0.003), 3–5 times (p = 0.004), and >5 times (p = 0.009) were more likely to be aware of ODS than others. Our findings indicated that pregnant women have little knowledge of ODS, which makes it difficult for them to take prompt decisions to seek emergency obstetric care. Thus, strategies to increase the knowledge of pregnant women about obstetrical danger signs by healthcare providers during prenatal consultations (antenatal care) must be developed to improve their rapid decision-making skills during pregnancy, labor, and postpartum.
Insights into the Molecular Basis of Huanglongbing Tolerance in Persian Lime (Citrus latifolia Tan.) through a Transcriptomic Approach
Authors: Estrella-Maldonado Humberto , González-Cruz Carlos , Matilde-Hernández Cristian , Adame-García Jacel , Santamaría Jorge M. , Santillán-Mendoza Ricardo , Flores-de la Rosa Felipe Roberto
Published: April 2023

Huanglongbing (HLB) is a vascular disease of Citrus caused by three species of the α-proteobacteria “Candidatus Liberibacter”, with “Candidatus Liberibacter asiaticus” (CLas) being the most widespread and the one causing significant economic losses in citrus-producing regions worldwide. However, Persian lime (Citrus latifolia Tanaka) has shown tolerance to the disease. To understand the molecular mechanisms of this tolerance, transcriptomic analysis of HLB was performed using asymptomatic and symptomatic leaves. RNA-Seq analysis revealed 652 differentially expressed genes (DEGs) in response to CLas infection, of which 457 were upregulated and 195 were downregulated. KEGG analysis revealed that after CLas infection, some DEGs were present in the plant–pathogen interaction and in the starch and sucrose metabolism pathways. DEGs present in the plant–pathogen interaction pathway suggests that tolerance against HLB in Persian lime could be mediated, at least partly, by the ClRSP2 and ClHSP90 genes. Previous reports documented that RSP2 and HSP90 showed low expression in susceptible citrus genotypes. Regarding the starch and sucrose metabolism pathways, some genes were identified as being related to the imbalance of starch accumulation. On the other hand, eight biotic stress-related genes were selected for further RT-qPCR analysis to validate our results. RT-qPCR results confirmed that symptomatic HLB leaves had high relative expression levels of the ClPR1, ClNFP, ClDR27, and ClSRK genes, whereas the ClHSL1, ClRPP13, ClPDR1, and ClNAC genes were expressed at lower levels than those from HLB asymptomatic leaves. Taken together, the present transcriptomic analysis contributes to the understanding of the CLas-Persian lime interaction in its natural environment and may set the basis for developing strategies for the integrated management of this important Citrus disease through the identification of blanks for genetic improvement.
Lipopolysaccharides: Regulated Biosynthesis and Structural Diversity
Authors: Raina Satish
Published: April 2023

The cell envelope of Gram-negative bacteria contains two distinct membranes, an inner (IM) and an outer (OM) membrane, separated by the periplasm, a hydrophilic compartment that includes a thin layer of peptidoglycan [...]
Cannabis Use and Endocannabinoid Receptor Genes: A Pilot Study on Their Interaction on Brain Activity in First-Episode Psychosis
Authors: Oscoz-Irurozqui Maitane , Almodóvar-Payá Carmen , Guardiola-Ripoll Maria , Guerrero-Pedraza Amalia , Hostalet Noemí , Salvador Raymond , Carrión Maria Isabel , Maristany Teresa , Pomarol-Clotet Edith , Fatjó-Vilas Mar
Published: April 2023

The role of both cannabis use and genetic background has been shown in the risk for psychosis. However, the effect of the interplay between cannabis and variability at the endocannabinoid receptor genes on the neurobiological underpinnings of psychosis remains inconclusive. Through a case-only design, including patients with a first-episode of psychosis (n = 40) classified as cannabis users (50%) and non-users (50%), we aimed to evaluate the interaction between cannabis use and common genetic variants at the endocannabinoid receptor genes on brain activity. Genetic variability was assessed by genotyping two Single Nucleotide Polymorphisms (SNP) at the cannabinoid receptor type 1 gene (CNR1; rs1049353) and cannabinoid receptor type 2 gene (CNR2; rs2501431). Functional Magnetic Resonance Imaging (fMRI) data were obtained while performing the n-back task. Gene × cannabis interaction models evidenced a combined effect of CNR1 and CNR2 genotypes and cannabis use on brain activity in different brain areas, such as the caudate nucleus, the cingulate cortex and the orbitofrontal cortex. These findings suggest a joint role of cannabis use and cannabinoid receptor genetic background on brain function in first-episode psychosis, possibly through the impact on brain areas relevant to the reward circuit.
Impact of Adipose Tissue Depot Harvesting Site on the Multilineage Induction Capacity of Male Rat Adipose-Derived Mesenchymal Stem Cells: An In Vitro Study
Authors: El-Husseiny Hussein M. , Kaneda Masahiro , Mady Eman A. , Yoshida Tadashi , Doghish Ahmed S. , Tanaka Ryou
Published: April 2023

Recently, substantial attention has been paid toward adipose-derived mesenchymal stem cells (AdMSCs) as a potential therapy in tissue engineering and regenerative medicine applications. Rat AdMSCs (r-AdMSCs) are frequently utilized. However, the influence of the adipose depot site on the multilineage differentiation potential of the r-AdMSCs is still ambiguous. Hence, the main objective of this study was to explore the influence of the adipose tissue harvesting location on the ability of r-AdMSCs to express the stem-cell-related markers and pluripotency genes, as well as their differentiation capacity, for the first time. Herein, we have isolated r-AdMSCs from the inguinal, epididymal, peri-renal, and back subcutaneous fats. Cells were compared in terms of their phenotype, immunophenotype, and expression of pluripotency genes using RT-PCR. Additionally, we investigated their potential for multilineage (adipogenic, osteogenic, and chondrogenic) induction using special stains confirmed by the expression of the related genes using RT-qPCR. All cells could positively express stem cell marker CD 90 and CD 105 with no significant in-between differences. However, they did not express the hematopoietic markers as CD 34 and CD 45. All cells could be induced successfully. However, epididymal and inguinal cells presented the highest capacity for adipogenic and osteogenic differentiation (21.36-fold and 11.63-fold for OPN, 29.69-fold and 26.68-fold for BMP2, and 37.67-fold and 22.35-fold for BSP, respectively, in epididymal and inguinal cells (p < 0.0001)). On the contrary, the subcutaneous cells exhibited a superior potential for chondrogenesis over the other sites (8.9-fold for CHM1 and 5.93-fold for ACAN, (p < 0.0001)). In conclusion, the adipose tissue harvesting site could influence the differentiation capacity of the isolated AdMSCs. To enhance the results of their employment in various regenerative cell-based therapies, it is thus vital to take the collection site selection into consideration.
Redox and Nucleophilic Reactions of Naphthoquinones with Small Thiols and Their Effects on Oxidization of H2S to Inorganic and Organic Hydropolysulfides and Thiosulfate
Authors: Olson Kenneth R. , Clear Kasey J. , Gao Yan , Ma Zhilin , Cieplik Nathaniel M. , Fiume Alyssa R. , Gaziano Dominic J. , Kasko Stephen M. , Luu Jennifer , Pfaff Ella , Travlos Anthony , Velander Cecilia , Wilson Katherine J. , Edwards Elizabeth D. , Straub Karl D. , Wu Gang
Published: April 2023

Naphthoquinone (1,4-NQ) and its derivatives (NQs, juglone, plumbagin, 2-methoxy-1,4-NQ, and menadione) have a variety of therapeutic applications, many of which are attributed to redox cycling and the production of reactive oxygen species (ROS). We previously demonstrated that NQs also oxidize hydrogen sulfide (H2S) to reactive sulfur species (RSS), potentially conveying identical benefits. Here we use RSS-specific fluorophores, mass spectroscopy, EPR and UV-Vis spectrometry, and oxygen-sensitive optodes to examine the effects of thiols and thiol-NQ adducts on H2S-NQ reactions. In the presence of glutathione (GSH) and cysteine (Cys), 1,4-NQ oxidizes H2S to both inorganic and organic hydroper-/hydropolysulfides (R2Sn, R=H, Cys, GSH; n = 2–4) and organic sulfoxides (GSnOH, n = 1, 2). These reactions reduce NQs and consume oxygen via a semiquinone intermediate. NQs are also reduced as they form adducts with GSH, Cys, protein thiols, and amines. Thiol, but not amine, adducts may increase or decrease H2S oxidation in reactions that are both NQ- and thiol-specific. Amine adducts also inhibit the formation of thiol adducts. These results suggest that NQs may react with endogenous thiols, including GSH, Cys, and protein Cys, and that these adducts may affect both thiol reactions as well as RSS production from H2S.
Targeted Large-Volume Lymphocyte Removal Using Magnetic Nanoparticles in Blood Samples of Patients with Chronic Lymphocytic Leukemia: A Proof-of-Concept Study
Authors: Janker Stefanie , Doswald Simon , Schimmer Roman R. , Schanz Urs , Stark Wendelin J. , Schläpfer Martin , Beck-Schimmer Beatrice
Published: April 2023

In the past, our research group was able to successfully remove circulating tumor cells with magnetic nanoparticles. While these cancer cells are typically present in low numbers, we hypothesized that magnetic nanoparticles, besides catching single cells, are also capable of eliminating a large number of tumor cells from the blood ex vivo. This approach was tested in a small pilot study in blood samples of patients suffering from chronic lymphocytic leukemia (CLL), a mature B-cell neoplasm. Cluster of differentiation (CD) 52 is a ubiquitously expressed surface antigen on mature lymphocytes. Alemtuzumab (MabCampath®) is a humanized, IgG1κ, monoclonal antibody directed against CD52, which was formerly clinically approved for treating chronic lymphocytic leukemia (CLL) and therefore regarded as an ideal candidate for further tests to develop new treatment options. Alemtuzumab was bound onto carbon-coated cobalt nanoparticles. The particles were added to blood samples of CLL patients and finally removed, ideally with bound B lymphocytes, using a magnetic column. Flow cytometry quantified lymphocyte counts before, after the first, and after the second flow across the column. A mixed effects analysis was performed to evaluate removal efficiency. p < 0.05 was defined as significant. In the first patient cohort (n = 10), using a fixed nanoparticle concentration, CD19-positive B lymphocytes were reduced by 38% and by 53% after the first and the second purification steps (p = 0.002 and p = 0.005), respectively. In a second patient cohort (n = 11), the nanoparticle concentration was increased, and CD19-positive B lymphocytes were reduced by 44% (p < 0.001) with no further removal after the second purification step. In patients with a high lymphocyte count (>20 G/L), an improved efficiency of approximately 20% was observed using higher nanoparticle concentrations. A 40 to 50% reduction of B lymphocyte count using alemtuzumab-coupled carbon-coated cobalt nanoparticles is feasible, also in patients with a high lymphocyte count. A second purification step did not further increase removal. This proof-of-concept study demonstrates that such particles allow for the targeted extraction of larger amounts of cellular blood components and might offer new treatment options in the far future.
Interleukin 26 Induces Macrophage IL-9 Expression in Rheumatoid Arthritis
Authors: Wang Yi-Hsun , Peng Yi-Jen , Liu Feng-Cheng , Lin Gu-Jiun , Huang Shing-Hwa , Sytwu Huey-Kang , Cheng Chia-Pi
Published: April 2023

Rheumatoid arthritis (RA) is an autoimmune disease with chronic inflammation, bone erosion, and joint deformation. Synovial tissue in RA patients is full of proinflammatory cytokines and infiltrated immune cells, such as T help (Th) 9, Th17, macrophages, and osteoclasts. Recent reports emphasized a new member of the interleukin (IL)-10 family, IL-26, an inducer of IL-17A that is overexpressed in RA patients. Our previous works found that IL-26 inhibits osteoclastogenesis and conducts monocyte differentiation toward M1 macrophages. In this study, we aimed to clarify the effect of IL-26 on macrophages linking to Th9 and Th17 in IL-9 and IL-17 expression and downstream signal transduction. Murine and human macrophage cell lines and primary culture cells were used and stimulated by IL26. Cytokines expressions were evaluated by flow cytometry. Signal transduction and transcription factors expression were detected by Western blot and real time-PCR. Our results show that IL-26 and IL-9 colocalized in macrophage in RA synovium. IL-26 directly induces macrophage inflammatory cytokines IL-9 and IL-17A expression. IL-26 increases the IL-9 and IL-17A upstream mechanisms IRF4 and RelB expression. Moreover, the AKT-FoxO1 pathway is also activated by IL-26 in IL-9 and IL-17A expressing macrophage. Blockage of AKT phosphorylation enhances IL-26 stimulating IL-9-producing macrophage cells. In conclusion, our results support that IL-26 promotes IL-9- and IL-17-expressing macrophage and might initiate IL-9- and IL-17-related adaptive immunity in rheumatoid arthritis. Targeting IL-26 may a potential therapeutic strategy for rheumatoid arthritis or other IL-9 plus IL-17 dominant diseases.
Intimal Sarcoma with MDM2/CDK4 Amplification and p16 Overexpression: A Review of Histological Features in Primary Tumor and Xenograft, with Immunophenotype and Molecular Profiling
Authors: Giner Francisco , Machado Isidro , Rubio-Martínez Luis Alberto , López-Guerrero José Antonio , Claramunt-Alonso Reyes , Navarro Samuel , Ferrández Antonio , Mayordomo-Aranda Empar , Llombart-Bosch Antonio
Published: April 2023

Intimal sarcomas (IS) are rare malignant mesenchymal tumors arising in large blood vessels of the systemic and pulmonary circulation and also in the heart. They are morphologically similar to other spindle cell, poorly differentiated sarcomas. The prognosis is poor and depends mainly on surgical options. Three cases of IS were collected from two institutions. Clinical data were retrieved and histological study was performed. A wide immunohistochemical panel was analyzed. FISH of MDM2 gene was performed, and a molecular study with NGS was implemented in all cases. The mean age of our cases was 54 years. Histologically, the tumors presented a diffuse growth pattern with heterogeneous atypical epithelioid or spindle cells and extensive thrombosed areas. All cases presented intense immunoexpression for MDM2, CDK4, CD117, c-myc, PDGFRA, and p16. PDGFRA, HTERT, and pan-TRK gained expression, while p16 lost intensity, being weaker in both the local recurrences and xenografts. The three cases showed amplification of MDM2 by FISH. NGS analysis revealed amplifications in the CDK4, PDGFRA, and KIT genes, together with BRAF mutation and KRAS amplification. P16 was expressed in all cases, losing intensity in local recurrence and xenografts. Two new alterations, a BRAF mutation and a KRAS amplification, were detected by NGS in different tumors, opening up new therapeutic options for these patients.
Characterization of Porcine Ocular Surface Epithelial Microenvironment
Authors: Polisetti Naresh , Martin Gottfried , Cristina Schmitz Heidi R. , Schlötzer-Schrehardt Ursula , Schlunck Günther , Reinhard Thomas
Published: April 2023

The porcine ocular surface is used as a model of the human ocular surface; however, a detailed characterization of the porcine ocular surface has not been documented. This is due, in part, to the scarcity of antibodies produced specifically against the porcine ocular surface cell types or structures. We performed a histological and immunohistochemical investigation on frozen and formalin-fixed, paraffin-embedded ocular surface tissue from domestic pigs using a panel of 41 different antibodies related to epithelial progenitor/differentiation phenotypes, extracellular matrix and associated molecules, and various niche cell types. Our observations suggested that the Bowman’s layer is not evident in the cornea; the deep invaginations of the limbal epithelium in the limbal zone are analogous to the limbal interpalisade crypts of human limbal tissue; and the presence of goblet cells in the bulbar conjunctiva. Immunohistochemistry analysis revealed that the epithelial progenitor markers cytokeratin (CK)15, CK14, p63α, and P-cadherin were expressed in both the limbal and conjunctival basal epithelium, whereas the basal cells of the limbal and conjunctival epithelium did not stain for CK3, CK12, E-cadherin, and CK13. Antibodies detecting marker proteins related to the extracellular matrix (collagen IV, Tenascin-C), cell–matrix adhesion (β-dystroglycan, integrin α3 and α6), mesenchymal cells (vimentin, CD90, CD44), neurons (neurofilament), immune cells (HLA-ABC; HLA-DR, CD1, CD4, CD14), vasculature (von Willebrand factor), and melanocytes (SRY-homeobox-10, human melanoma black-45, Tyrosinase) on the normal human ocular surface demonstrated similar immunoreactivity on the normal porcine ocular surface. Only a few antibodies (directed against N-cadherin, fibronectin, agrin, laminin α3 and α5, melan-A) appeared unreactive on porcine tissues. Our findings characterize the main immunohistochemical properties of the porcine ocular surface and provide a morphological and immunohistochemical basis useful to research using porcine models. Furthermore, the analyzed porcine ocular structures are similar to those of humans, confirming the potential usefulness of pig eyes to study ocular surface physiology and pathophysiology.
Zonulin as a Potential Therapeutic Target in Microbiota-Gut-Brain Axis Disorders: Encouraging Results and Emerging Questions
Authors: Veres-Székely Apor , Szász Csenge , Pap Domonkos , Szebeni Beáta , Bokrossy Péter , Vannay Ádám
Published: April 2023

The relationship between dysbiosis and central nervous diseases has been proved in the last 10 years. Microbial alterations cause increased intestinal permeability, and the penetration of bacterial fragment and toxins induces local and systemic inflammatory processes, affecting distant organs, including the brain. Therefore, the integrity of the intestinal epithelial barrier plays a central role in the microbiota–gut–brain axis. In this review, we discuss recent findings on zonulin, an important tight junction regulator of intestinal epithelial cells, which is assumed to play a key role in maintaining of the blood–brain barrier function. In addition to focusing on the effect of microbiome on intestinal zonulin release, we also summarize potential pharmaceutical approaches to modulate zonulin-associated pathways with larazotide acetate and other zonulin receptor agonists or antagonists. The present review also addresses the emerging issues, including the use of misleading nomenclature or the unsolved questions about the exact protein sequence of zonulin.
DoMYB5 and DobHLH24, Transcription Factors Involved in Regulating Anthocyanin Accumulation in Dendrobium officinale
Authors: Yang Kun , Hou Yibin , Wu Mei , Pan Qiuyu , Xie Yilong , Zhang Yusen , Sun Fenghang , Zhang Zhizhong , Wu Jinghua
Published: April 2023

As a kind of orchid plant with both medicinal and ornamental value, Dendrobium officinale has garnered increasing research attention in recent years. The MYB and bHLH transcription factors play important roles in the synthesis and accumulation of anthocyanin. However, how MYB and bHLH transcription factors work in the synthesis and accumulation of anthocyanin in D. officinale is still unclear. In this study, we cloned and characterized one MYB and one bHLH transcription factor, namely, D. officinale MYB5 (DoMYB5) and D. officinaleb bHLH24 (DobHLH24), respectively. Their expression levels were positively correlated with the anthocyanin content in the flowers, stems, and leaves of D. officinale varieties with different colors. The transient expression of DoMYB5 and DobHLH24 in D. officinale leaf and their stable expression in tobacco significantly promoted the accumulation of anthocyanin. Both DoMYB5 and DobHLH24 could directly bind to the promoters of D. officinale CHS (DoCHS) and D. officinale DFR (DoDFR) and regulate DoCHS and DoDFR expression. The co-transformation of the two transcription factors significantly enhanced the expression levels of DoCHS and DoDFR. DoMYB5 and DobHLH24 may enhance the regulatory effect by forming heterodimers. Drawing on the results of our experiments, we propose that DobHLH24 may function as a regulatory partner by interacting directly with DoMYB5 to stimulate anthocyanin accumulation in D. officinale.
Quantifying Functional Impairment of ABCA3 Variants Associated with Interstitial Lung Disease
Authors: Yang Xiaohua , Rapp Christina K. , Li Yang , Forstner Maria , Griese Matthias
Published: April 2023

ATP-binding cassette subfamily A member 3 (ABCA3) is a lipid transporter within alveolar type II cells. Patients with bi-allelic variants in ABCA3 may suffer from a variable severity of interstitial lung disease. We characterized and quantified ABCA3 variants’ overall lipid transport function by assessing the in vitro impairment of its intracellular trafficking and pumping activity. We expressed the results relative to the wild type, integrated the quantitative readouts from eight different assays and used newly generated data combined with previous results to correlate the variants’ function and clinical phenotype. We differentiated normal (within 1 normalized standard deviation (nSD) of the wild-type mean), impaired (within 1 to 3 nSD) and defective (beyond 3 nSD) variants. The transport of phosphatidylcholine from the recycling pathway into ABCA3+ vesicles proved sensitive to the variants’ dysfunction. The sum of the quantitated trafficking and pumping predicted a clinical outcome. More than an approximately 50% loss of function was associated with considerable morbidity and mortality. The in vitro quantification of ABCA3 function enables detailed variant characterization, substantially improves the phenotype prediction of genetic variants and possibly supports future treatment decisions.
Magnetic Field Effect in Bimolecular Rate Constant of Radical Recombination
Authors: Doktorov Alexander B. , Lukzen Nikita N.
Published: April 2023

The influence of magnetic fields on chemical reactions, including biological ones, has been and still is a topical subject in the field of scientific research. Experimentally discovered and theoretically substantiated magnetic and spin effects in chemical radical reactions form the basis of research in the field of spin chemistry. In the present work, the effect of a magnetic field on the rate constant of the bimolecular spin-selective recombination of radicals in the bulk of a solution is considered theoretically for the first time, taking into account the hyperfine interaction of radical spins with their magnetic nuclei. In addition, the paramagnetic relaxation of unpaired spins of the radicals and the non-equality of their g-factors that also influence the recombination process are taken into account. It is found that the reaction rate constant can vary in magnetic field from a few to half a dozen percent, depending on the relative diffusion coefficient of radicals, which is determined by the solution viscosity. It is shown that the consideration of hyperfine interactions gives rise to the presence of resonances in the dependence of the rate constant on the magnetic field. The magnitudes of the magnetic fields of these resonances are determined by the hyperfine coupling constants and difference in the g-factors of the recombining radicals. Analytical expressions for the reaction rate constant of the bulk recombination for magnetic fields larger than hfi (hyperfine interaction) constants are obtained. In general, it is shown for the first time that accounting for hyperfine interactions of radical spins with magnetic nuclei significantly affects the dependence of the reaction rate constant of the bulk radical recombination on the magnetic field.
Valorisation of Whey Permeate in Sequential Bioprocesses towards Value-Added Products–Optimisation of Biphasic and Classical Batch Cultures of Kluyveromyces marxianus
Authors: Drężek Karolina , Sobczyk Maria Krystyna , Kállai Zoltán , Detman Anna , Bardadyn Paula , Mierzejewska Jolanta
Published: April 2023

Whey permeate is categorised as hazardous wastewater for aquatic environments, mainly due to its high lactose content. Therefore, it must be valorised before being released into the environment. One pathway for whey permeate management is its use in biotechnological processes. Herein, we present roads for whey permeate valorisation with the K. marxianus WUT240 strain. The established technology is based on two bioprocesses. During first, 2.5 g/L 2-phenylethanol and fermented plant oils enriched with different flavourings are obtained after 48 h biphasic cultures at 30 °C. The second process leads to a maximum of 75 g ethanol/L (YP/S = 0.53 g/g) after 96 h at 30 °C. Moreover, established whey permeate valorisation pathways reduced its biochemical oxygen demand and chemical oxygen demand values by 12- to 3-fold, respectively. Together, the present study reports a complete, effective, and environmentally friendly whey permeate management strategy while simultaneously enabling the acquisition of valuable compounds with substantial application potential.
Genome-Wide Identification and Expression Analysis of NCED Gene Family in Pear and Its Response to Exogenous Gibberellin and Paclobutrazol
Authors: Liu Jinming , Yuan Xing , Quan Shaowen , Zhang Meng , Kang Chao , Guo Caihua , Zhang Zhongrong , Niu Jianxin
Published: April 2023

The 9-cis-epoxycarotenoid dioxygenase (NCED) is a key enzyme for the process of ABA synthesis that plays key roles in a variety of biological processes. In the current investigation, genome-wide identification and comprehensive analysis of the NCED gene family in ‘Kuerle Xiangli’ (Pyrus sinkiangensis Yu) were conducted using the pear genomic sequence. In total, nineteen members of PbNCED genes were identified from the whole genome of pear, which are not evenly distributed over the scaffolds, and most of which were focussed in the chloroplasts. Sequence analysis of promoters showed many cis-regulatory elements, which presumably responded to phytohormones such as abscisic acid, auxin, etc. Synteny block indicated that the PbNCED genes have experienced strong purifying selection. Multiple sequence alignment demonstrated that these members are highly similar and conserved. In addition, we found that PbNCED genes were differentially expressed in various tissues, and three PbNCED genes (PbNCED1, PbNCED2, and PbNCED13) were differentially expressed in response to exogenous Gibberellin (GA3) and Paclobutrazol (PP333). PbNCED1 and PbNCED13 positively promote ABA synthesis in sepals after GA3 and PP333 treatment, whereas PbNCED2 positively regulated ABA synthesis in ovaries after GA3 treatment, and PbNCED13 positively regulated ABA synthesis in the ovaries after PP333 treatment. This study was the first genome-wide report of the pear NCED gene family, which could improve our understanding of pear NCED proteins and provide a solid foundation for future cloning and functional analyses of this gene family. Meanwhile, our results also give a better understanding of the important genes and regulation pathways related to calyx abscission in ‘Kuerle Xiangli’.
Linderone Isolated from Lindera erythrocarpa Exerts Antioxidant and Anti-Neuroinflammatory Effects via NF-κB and Nrf2 Pathways in BV2 and HT22 Cells
Authors: Liu Zhiming , Yoon Chi-Su , Lee Hwan , Lee Hyeong-Kyu , Lee Dong-Sung
Published: April 2023

Linderone is a major compound in Lindera erythrocarpa and exhibits anti-inflammatory effects in BV2 cells. This study investigated the neuroprotective effects and mechanisms of linderone action in BV2 and HT22 cells. Linderone suppressed lipopolysaccharide (LPS)-induced inducible nitric oxide synthase, cyclooxygenase-2, and pro-inflammatory cytokines (e.g., tumor necrosis factor alpha, interleukin-6, and prostaglandin E-2) in BV2 cells. Linderone treatment also inhibited the LPS-induced activation of p65 nuclear factor-kappa B, protecting against oxidative stress in glutamate-stimulated HT22 cells. Furthermore, linderone activated the translocation of nuclear factor E2-related factor 2 and induces the expression of heme oxygenase-1. These findings provided a mechanistic explanation of the antioxidant and anti-neuroinflammatory effects of linderone. In conclusion, our study demonstrated the therapeutic potential of linderone in neuronal diseases.
Photochemically Aided Arteriovenous Fistula Creation to Accelerate Fistula Maturation
Authors: He Yong , Anderson Blake , Hu Qiongyao , Hayes RB , Huff Kenji , Isaacson Jim , Warner Kevin S. , Hauser Hank , Greenberg Myles , Chandra Venita , Kauser Katalin , Berceli Scott A.
Published: April 2023

Rates of arteriovenous fistula maturation failure are still high, especially when suboptimal size veins are used. During successful maturation, the vein undergoes lumen dilatation and medial thickening, adapting to the increased hemodynamic forces. The vascular extracellular matrix plays an important role in regulating these adaptive changes and may be a target for promoting fistula maturation. In this study, we tested whether a device-enabled photochemical treatment of the vein prior to fistula creation facilitates maturation. Sheep cephalic veins were treated using a balloon catheter coated by a photoactivatable molecule (10-8-10 Dimer) and carrying an internal light fiber. As a result of the photochemical reaction, new covalent bonds were created during light activation among oxidizable amino acids of the vein wall matrix proteins. The treated vein lumen diameter and media area became significantly larger than the contralateral control fistula vein at 1 week (p = 0.035 and p = 0.034, respectively). There was also a higher percentage of proliferating smooth muscle cells in the treated veins than in the control veins (p = 0.029), without noticeable intimal hyperplasia. To prepare for the clinical testing of this treatment, we performed balloon over-dilatation of isolated human veins and found that veins can tolerate up to 66% overstretch without notable histological damage.
Copper Accumulation Efficiency in Different Recombinant Microorganism Strains Available for Bioremediation of Heavy Metal-Polluted Waters
Authors: Vulpe Constantina Bianca , Matica Mariana Adina , Kovačević Renata , Dascalu Daniela , Stevanovic Zoran , Isvoran Adriana , Ostafe Vasile , Menghiu Gheorghița
Published: April 2023

The aim of this research was to investigate the bioremediation conditions of copper in synthetic water. In the present study, copper ions accumulation efficiency was determined using various genetically modified strains of Saccharomyces cerevisiae (EBY100, INVSc1, BJ5465, and GRF18), Pichia pastoris (X-33, KM71H), Escherichia coli (XL10 Gold, DH5α, and six types of BL21 (DE3)), and Escherichia coli BL21 (DE3) OverExpress expressing two different peroxidases. Viability tests of yeast and bacterial strains showed that bacteria are viable at copper concentrations up to 2.5 mM and yeasts up to 10 mM. Optical emission spectrometry with inductively coupled plasma analysis showed that the tolerance of bacterial strains on media containing 1 mM copper was lower than the tolerance of yeast strains at the same copper concentration. The E. coli BL21 RIL strain had the best copper accumulation efficiency (4.79 mg/L of culture normalized at an optical density of 1.00), which was 1250 times more efficient than the control strain. The yeast strain S. cerevisiae BJ5465 was the most efficient in copper accumulation out of a total of six yeast strains used, accumulating over 400 times more than the negative control strain. In addition, E. coli cells that internally expressed recombinant peroxidase from Thermobifida fusca were able to accumulate 400-fold more copper than cells that produced periplasmic recombinant peroxidases.
Prognostic and Predictive Biomarkers in the Era of Immunotherapy for Lung Cancer
Authors: Pabst Lucile , Lopes Sébastien , Bertrand Basil , Creusot Quentin , Kotovskaya Maria , Pencreach Erwan , Beau-Faller Michèle , Mascaux Céline
Published: April 2023

The therapeutic algorithm of lung cancer has recently been revolutionized by the emergence of immune checkpoint inhibitors. However, an objective and durable response rate remains low with those recent therapies and some patients even experience severe adverse events. Prognostic and predictive biomarkers are therefore needed in order to select patients who will respond. Nowadays, the only validated biomarker is the PD-L1 expression, but its predictive value remains imperfect, and it does not offer any certainty of a sustained response to treatment. With recent progresses in molecular biology, genome sequencing techniques, and the understanding of the immune microenvironment of the tumor and its host, new molecular features have been highlighted. There are evidence in favor of the positive predictive value of the tumor mutational burden, as an example. From the expression of molecular interactions within tumor cells to biomarkers circulating in peripheral blood, many markers have been identified as associated with the response to immunotherapy. In this review, we would like to summarize the latest knowledge about predictive and prognostic biomarkers of immune checkpoint inhibitors efficacy in order to go further in the field of precision immuno-oncology.
Disruption of Irisin Dimerization by FDA-Approved Drugs: A Computational Repurposing Approach for the Potential Treatment of Lipodystrophy Syndromes
Authors: Flori Lorenzo , Brogi Simone , Sirous Hajar , Calderone Vincenzo
Published: April 2023

In this paper, we present the development of a computer-based repurposing approach to identify FDA-approved drugs that are potentially able to interfere with irisin dimerization. It has been established that altered levels of irisin dimers are a pure hallmark of lipodystrophy (LD) syndromes. Accordingly, the identification of compounds capable of slowing down or precluding the irisin dimers’ formation could represent a valuable therapeutic strategy in LD. Combining several computational techniques, we identified five FDA-approved drugs with satisfactory computational scores (iohexol, XP score = −7.70 kcal/mol, SP score = −5.5 kcal/mol, ΔGbind = −61.47 kcal/mol, ΔGbind (average) = −60.71 kcal/mol; paromomycin, XP score = −7.23 kcal/mol, SP score = −6.18 kcal/mol, ΔGbind = −50.14 kcal/mol, ΔGbind (average) = −49.13 kcal/mol; zoledronate, XP score = −6.33 kcal/mol, SP score = −5.53 kcal/mol, ΔGbind = −32.38 kcal/mol, ΔGbind (average) = −29.42 kcal/mol; setmelanotide, XP score = −6.10 kcal/mol, SP score = −7.24 kcal/mol, ΔGbind = −56.87 kcal/mol, ΔGbind (average) = −62.41 kcal/mol; and theophylline, XP score = −5.17 kcal/mol, SP score = −5.55 kcal/mol, ΔGbind = −33.25 kcal/mol, ΔGbind (average) = −35.29 kcal/mol) that are potentially able to disrupt the dimerization of irisin. For this reason, they deserve further investigation to characterize them as irisin disruptors. Remarkably, the identification of drugs targeting this process can offer novel therapeutic opportunities for the treatment of LD. Furthermore, the identified drugs could provide a starting point for a repositioning approach, synthesizing novel analogs with improved efficacy and selectivity against the irisin dimerization process.
Immobilization on Polyethylenimine and Chitosan Sorbents Modulates the Production of Valuable Fatty Acids by the Chlorophyte Lobosphaera sp. IPPAS C-2047
Authors: Vasilieva Svetlana , Shibzukhova Karina , Solovchenko Alexei , Chivkunova Olga , Antipova Christina , Morozov Alexey , Lobakova Elena
Published: April 2023

Green microalgae, including those from the genus Lobosphaera, are exploited in various fields of biotechnology to obtain valuable fatty acids (e.g., arachidonic acid (C20:4, ARA)) for the production of infant formulae, food and feed additives. In nature, microalgae frequently exist in naturally immobilized state (as biofilms) with a limited cell division rate and increased stress resilience. In the fields of biotechnology, immobilization of microalgae on artificial cell carriers simplifies biomass harvesting and increases culture robustness and productivity. The choice of a suitable cell carrier is central to biotechnology involving immobilized cultures. Cell carriers based on the natural amine-containing polymer chitosan and synthetic polyethylenimine (PEI) are promising candidates for immobilization of phototrophic microorganisms. This is the first report on the effects of immobilization on PEI and chitosan on the accumulation and composition of polyunsaturated fatty acids, including ARA, in Lobosphaera sp. IPPAS C-2047. Immobilization on PEI increased the ARA percentage in the total fatty acids and ARA accumulation by 72% and 81% compared to the suspended cells cultured in complete or nitrogen-deprived medium 14 days, respectively. Immobilization of Lobosphaera sp. on the chitosan-based carrier reduced the ARA percentage but increased oleic and α-linoleic acid percentages. The mechanisms of the effects of immobilization on the fatty acid profiles of the microalgae are discussed.