bluuebunny/crossref_metadata_2025 huggingface dataset:
Investigating the Synergistic Effects of Carvacrol and Citral-Edible Polysaccharide-Based Nanoemulgels on Shelf Life Extension of Chalkidiki Green Table Olives
Authors: Zaharioudakis Konstantinos , Salmas Constantinos E. , Andritsos Nikolaos D. , Leontiou Areti A. , Moschovas Dimitrios , Karydis-Messinis Andreas , Triantafyllou Eleni , Avgeropoulos Apostolos , Zafeiropoulos Nikolaos E. , Proestos Charalampos , Giannakas Aris E.
Published: November 2024

Modern bioeconomy and sustainability demands lead food technology in the development of novel biobased edible food preservatives. Herein, the development and characterization of novel polysaccharide (xanthan gum and kappa-carrageenan)-based nanoemulgels (NGs) enhanced with essential oil derivatives; pure citral (CT); pure carvacrol (CV); and various CT:CV ratios (25:75, 50:50, and 75:25) are presented. The obtained NGs are applied as active edible coatings for extending the shelf life of Protected Designation of Origin (PDO) green table olives of Chalkidiki. The zeta potential demonstrated the high stability of the treatments, while light scattering measurement and scanning electron microscopy images confirmed the <100 nm droplet size. EC50 indicated high antioxidant activity for all the tested samples. The fractional inhibitory concentration (FIC) confirmed the synergistic effect of NG with a CT:CV ratio at 50:50 against Staphylococcus aureus and at CT:CV ratios 25:75 and 75:25 against E. coli O157:H7. NG coatings with CT:CV ratios at 50:50 and at 25:75 effectively controlled the weight loss at 0.5%, maintained stable pH levels, and preserved the visual quality of green olives on day 21. The synergistic effect between CT and CV was confirmed as they reduced the spoilage microorganisms of yeasts and molds by 2-log [CFU/g] compared to the control and almost 1 log [CFU/g] difference from pure CT and CV-based NGs without affecting the growth of beneficial lactic acid bacteria crucial for fermentation. NGs with CT:CV ratios at 50:50 and at 25:75 demonstrated superior effectiveness in preventing discoloration and maintaining the main sensory attributes. Overall, shelf life extension was achieved in 21 compared to only 7 of the uncoated ones. Finally, this study demonstrates the potential of polysaccharide-based NGs in mixtures of CT and CV for the shelf life extension of fermented food products.
Improvement of Adsorption Capacity by Refined Encapsulating Method of Activated Carbon into the Hollow-Type Spherical Bacterial Cellulose Gels for Oral Absorbent
Authors: Hirai Aya , Sato Kaito , Hoshi Toru , Aoyagi Takao
Published: November 2024

To reduce the risk of adsorption of granular activated carbon (AC) in the gastrointestinal tract, we successfully prepared a hollow-type spherical bacterial cellulose gel encapsulated with AC (ACEG) and evaluated its pH tolerance and adsorption capacity. The bacterial cellulose gel membrane of ACEG features a three-dimensional mesh structure of cellulose fibers, allowing the selective permeation of substances based on their size. In this study, the preparation method of ACEGs was investigated, and the indole saturation adsorption capacity of the obtained gel was measured. We modified the gel culture nucleus gel from calcium alginate gel to agar gel, facilitating the encapsulation of previously challenging particles. The new preparation method used sodium hydroxide solution for sterilization and dissolution to remove the debris of Komagataeibacter xylinus, which was feared to remain in the bacterial cellulose membrane. This treatment was also confirmed to have no effect on the adsorption capacity of the AC powder. Therefore, this new preparation method is expected not only to improve the performance of ACEGs but also to be applied to a wide range of adsorbent-encapsulated hollow-type bacterial cellulose gels.
Characterization of HSP70 and HSP90 Gene Family in Takifugu fasciatus and Their Expression Profiles on Biotic and Abiotic Stresses Response
Authors: Zhang Wenwen , Qian Ziang , Ji Jie , Wang Tao , Yin Shaowu , Zhang Kai
Published: November 2024

Background: Heat shock proteins (HSPs) play crucial roles in response to temperature changes and biotic stresses. However, the HSP gene family in the pufferfish (Takifugu fasciatus) herring has not been comprehensively investigated. Methods and Results: This study presents a systematic analysis of the HSP70 and HSP90 gene families in T. fasciatus, focusing on gene characterization, conserved structural domains, molecular evolutionary history, and expression patterns of the HSP gene family under stress conditions. The findings reveal that 16 HSP genes are evolutionarily conserved, while hspa4 and hsp90aa appear specific to teleost fish. HSP genes exhibit widespread expression across 11 examined tissues, with most demonstrating high expression levels in the heart, brain, and liver. Furthermore, T. fasciatus was subjected to cryogenic and biotic stresses, revealing distinct expression patterns of HSPs under various stress conditions. The response of HSPs to cold stress and Aeromonas hydrophila infection was sustained. In contrast, gene expression of HSPs significantly changed only in the pre-infection period following Ichthyophthirius multifiliis infection, gradually returning to normal levels in the later stages. Conclusions: These experimental results provide a foundation for further in-depth investigations into the characteristics and functions of HSPs in T. fasciatus.
Efficiency of Primary Health Services in the Greek Public Sector: Evidence from Bootstrapped DEA/FDH Estimators
Authors: Flokou Angeliki , Aletras Vassilis H. , Miltiadis Chrysovalantis , Karaferis Dimitris Charalambos , Niakas Dimitris A.
Published: November 2024

Strengthening primary healthcare (PHC) is vital for enhancing efficiency and improving access, clinical outcomes, and population well-being. The World Health Organization emphasizes the role of effective PHC in reducing healthcare costs and boosting productivity. With growing healthcare demands and limited resources, efficient management is critical. Background/Objectives: Building on this point, this study aimed to evaluate the efficiency of PHC units across Greece, focusing on Health Centers (HCs) and Local Health Units (ToMYs). The objective was to assess their efficiency levels and identify factors contributing to observed inefficiencies. This study explores a novel research area by being the first to assess the efficiency of restructured primary healthcare facilities in Greece, utilizing 2019 data—the first year operational data became available for the newly established ToMY facilities following recent healthcare reforms. Methods: We applied a comprehensive suite of non-parametric methods, including Data Envelopment Analysis (DEA) under variable, constant, increasing, and decreasing returns to scale (VRS, CRS, IRS/NDRS, DRS/NIRS) assumptions, along with the Free Disposal Hull (FDH) model, all oriented toward output maximization. Efficiency scores were refined using bootstrapping to calculate 95% confidence intervals, and efficient units were ranked via the super-efficiency model. Outliers were identified and removed through the data cloud algorithm. For the first time at this scale, the final sample included the vast majority of PHC facilities in Greece—234 Health Centers and 94 Local Health Units—with inputs categorized into three human resource types: medical, nursing/paramedical, and administrative/other staff. Outputs encompassed scheduled visits, emergency visits, and pharmaceutical prescription visits. This diverse and comprehensive application of DEA methods represents a novel approach to evaluating PHC efficiency in Greece, with potential relevance to broader healthcare contexts. Results: The analysis revealed significant inefficiencies and differences in technical efficiency between HCs and ToMYs. HCs could nearly double their outputs (VRS score: 1.92), while ToMYs could increase theirs by 58% (VRS score: 1.58). Scale efficiency scores were closer, with HCs slightly more aligned with their optimal scale (1.17 vs. 1.20 for ToMYs). Conclusions: There is significant potential to improve efficiency in PHC, with variations depending on unit characteristics and regional differences. This evaluation provides a foundation for policymakers to identify areas for improvement and enhance the overall performance of healthcare services in Greece.
Factors Influencing Cucurbitacin-E-Glycoside Content in Bitter Hawkesbury Watermelon as Potential Synergist in Cucurbit Pest Management
Authors: Wallingford Anna , Hernandez Christopher , Halaweish Fathi , Ostlund Trevor , Short Brent , Weber Donald C.
Published: November 2024

Bitter Hawkesbury watermelon (BHW) Citrullus lanatus (Thunb.) Matsum. and Nakai (syn. Citrullus vulgaris Schad) contain high concentrations of cucurbitacin-E-glycoside (CEG), a compound that acts as an arrestant and feeding stimulant for diabroticine leaf beetles that are corn (maize) and cucurbit pests. Juice from BHW is used as feedstock to produce an insecticide synergist for improved chemical control of pests in cucurbit cropping systems. A positive linear relationship was observed between the CEG concentration of parent and offspring grown in open-pollinated field plots. However, subsequent experiments that explored the influence of parent and fruit maturity on CEG concentration did not confirm a relationship between accumulation patterns among offspring of half-sibling families. An effect of maturity was observed in that earlier harvested fruit had greater CEG concentrations than ripe or overripe fruit. In a field study, CIDETRAK L (active ingredient is BHW juice) was mixed with commonly used insecticides to enhance behavioral control of striped cucumber beetle Acalymma vittatum (F.) and squash vine borer Melittia cucurbitae (Harris). Equivalent control of A. vittatum and M. cucurbitae was observed on zucchini when treated with foliar applications of spinosad, acetamiprid, or lambda-cyhalothrin versus ground applications of the same products mixed with CIDETRAK L.
Nitrilase GiNIT from Gibberella intermedia Efficiently Degrades Nitriles Derived from Rapeseed Meal Glucosinolate
Authors: Li Han-Zhi , Liu Ming-Yu , Wang Yu-Yue , Luo Xue-Mei , Feng Jia-Xun , Zhao Shuai
Published: November 2024

Rapeseed meal is severely restricted in its utilization as unconventional animal feed due to anti-nutritive compounds, such as glucosinolate, that are degraded to toxic nitriles such as 3-butenenitrile and 4-pentenenitrile in animals. Few studies on nitrilases that can degrade glucosinolate-derived nitriles have been reported thus far. In the present study, a nitrilase gene GiNIT from Gibberella intermedia was over-expressed in Escherichia coli and the purified recombinant nitrilase rGiNIT showed specific activities of 134.48 U/mg and 122.16 U/mg when using 3-butenenitrile and 4-pentenenitrile as substrates at the optimal pH, 7.5, and temperature, 45 °C, which is the highest reported in the literature. The conversion of 3-butenenitrile and 4-pentenenitrile by rGiNIT reached 81.89% and 80.23% after hydrolysis for 15 min and 300 min, respectively. Site-directed mutagenesis and molecular docking analysis revealed that the catalytic ability of rGiNIT depended on the substrate binding pocket comprising 13 key amino acid residues. These results provide a potential enzyme resource for rapeseed meal detoxification and theoretical guidance for protein engineering.
Poly(Epsilon-Lysine) Dendrons Inhibit Proliferation in HER2-Overexpressing SKBR3 Breast Cancer Cells at Levels Higher than the Low-Expressing MDA-MB-231 Phenotype and Independently from the Presentation of HER2 Bioligands in Their Structure
Authors: Peregrino Giordana M. S. , Kudsiova Laila , Santin Matteo
Published: November 2024

Among the known breast cancers, the subtype with HER2 receptors-overexpressing cells is associated with a poor prognosis. The adopted monoclonal antibody Trastuzumab has improved clinical outcomes, but it is associated with drug resistance and relatively high costs. The present work adopted the peptide solid-phase synthesis method to synthesise branched poly(ε-lysine) peptide dendrons with 8 branching arms integrating, at their carboxy terminal molecular root, either an arginine or the HER2 receptor-binding sequence LSYCCK or the scramble sequence CSCLYK. These dendrons were synthesised in quantities higher than 100 mg/batch and with a purity exceeding 95%. When tested with two types of breast cancer cells, the dendrons led to levels of inhibition in the HER2 receptor-overexpressing breast cancer cells (SKBR3) comparable to Trastuzumab and higher than breast cancer cells with low receptor expression (MDA-MB-231) where inhibition was more moderate. Noticeably, the presence of the amino acid sequence LSYCCK at the dendron molecular root did not appear to produce any additional inhibitory effect. This was demonstrated also when the scramble sequence CSCLYK was integrated into the dendron and by the lack of any antiproliferative effect by the control linear target sequence. The specific inhibitory effect on proliferation was finally proven by the absence of cytotoxicity and normal expression of the cell migration marker N-Cadherin. Therefore, the present study shows the potential of poly(ε-lysine) dendrons as a cost-effective alternative to Trastuzumab in the treatment of HER2-positive breast cancer.
Observational Analyses of Ex Vivo Native American Platelet Responses
Authors: Goerger Krista , Caldwell Madison , Biermann Grace , Besh Fatima , Flickema Tanner , Patel Pramit , Abbott Karla , Holinstat Michael , Larson Mark K.
Published: November 2024

Platelet activation plays an essential role in clot formation to prevent blood loss following vascular damage. In pathologic conditions, platelet activation can lead to obstructive clots, disrupting blood flow and resulting in thrombosis. Native Americans suffer disproportionately from arterial disease and previous research has shown that Blacks are enriched in genetic polymorphisms that correlate with higher platelet reactivity contributing to an increased risk for thrombosis. Therefore, the current study sought to determine phenotypic variations in Native American platelet responses following stimulation with agonists, simulating vascular damage. Several donors from a small cohort of Native Americans showed atypical robust platelet aggregation when stimulated with submaximal concentrations of agonists. Further, when comparing α-granule secretion, a specific marker of platelet activation, Native Americans were more likely to have elevated responses to multiple agonist conditions of stimulation compared to Whites. Interestingly, there were no noticeable differences in integrin activation between Native Americans and Whites. Our study is the first to observe elevated Native American platelet responses compared to Whites, supporting further mechanistic studies and investigation of treatment approaches for the prevention of thrombosis.
In Vitro Evaluation of the Influence of Biosynthesized Calcium Oxide Nanoparticles on the Antibacterial Activity, pH, Microleakage and Cytotoxicity of Conventional Intracanal Medicaments
Authors: Kazi Fasiha Moin , Parvez Khurram , Asghar Asif , Akbar Shazia , Jawaed Noor-ul-Ain , Kumar Naresh , Palma Paulo J.
Published: November 2024

Intracanal medicaments are an important adjunct to the effective disinfection of the root canal system. However, conventional intracanal medicaments do not provide adequate protection against Enterococcus faecalis, which is the organism of interest in many cases of root canal failures. This study aimed to evaluate the influence of biosynthesized calcium oxide nanoparticles (CaO NPs) on the antibacterial activity, pH, microleakage and cytotoxicity of intracanal medicaments. CaO NPs were biosynthesized by the direct thermal decomposition of eggshells (EGS) and the reduction of calcium nitrate with papaya leaf extract (PLE). These nanoparticles were mixed with a proprietary calcium hydroxide powder in 10% and 25% (w/w) concentrations and blended in analytical-grade coconut oil to formulate the experimental medicaments. These were then evaluated for antibacterial activity, pH, microleakage and cytotoxicity at 1 day, 7 days and 15 days. A proprietary calcium hydroxide paste formulation (MX) was used as the control. Means and standard deviations were calculated and analyzed using repeated-measures ANOVA for pH and three-way ANOVA for the antibacterial effect, microleakage and cytotoxicity, followed by LSD post hoc analysis. Significant antibacterial activity was noted against Enterococcus faecalis at all times, with zones of inhibition (ZOI) up to 19.60 ± 2.30 mm. pH levels up to 13.13 ± 0.35 were observed for the experimental groups. Microleakage remained comparable to the control, while cytotoxicity was not observed in any of the groups at any time. Intracanal medicaments formulated with 10% and 25% (w/w) of biosynthesized CaO NPs could be promising candidates for the disinfection of the root canal system compared to conventional counterparts.
Multiple Aspects of Irritable Bowel Syndrome and the Role of the Immune System: An Overview of Systematic Reviews with a Focus on Polyphenols
Authors: Passacatini Lucia Carmela , Ilari Sara , Nucera Saverio , Scarano Federica , Macrì Roberta , Caminiti Rosamaria , Serra Maria , Oppedisano Francesca , Maiuolo Jessica , Palma Ernesto , Malafoglia Valentina , Tomino Carlo , Fini Massimo , Mollace Vincenzo , Muscoli Carolina
Published: November 2024

Irritable bowel syndrome (IBS) is a complex and often debilitating condition that significantly impacts the gastrointestinal system and the overall quality of life of those affected. IBS is characterized by a variety of distressing symptoms, including cramping, abdominal pain, and irregular bowel movements, underlined by an intricate interplay of immune system dysfunction in its pathology. Numerous studies highlight an increased cellular immune response, with elevated levels of proinflammatory cytokines, mucosal alterations due to immune imbalance, and visceral hypersensitivity. Notably, studies indicate increased levels of proinflammatory cytokines, immune imbalances that lead to mucosal changes, and heightened visceral sensitivity. The roles of effector and regulatory T cells are particularly intriguing, as their modification appears to amplify inflammation and may even contribute to autoimmune disorders. This overview of systematic reviews explores the connections between IBS and immune responses, with a focus on immune cell alterations and proliferation of lymphocytes and mast cells in affected individuals. Furthermore, we explore various aspects of IBS management, including its pharmacological approaches. A systematic search of PubMed and Web of Science yielded 676 articles, which were ultimately narrowed down to 9 key studies that met our inclusion criteria. These studies collectively underscore the activation of the immune system with the degranulation of the mast cells in patients with IBS, where the release of inflammatory mediators can compromise intestinal permeability, exacerbating symptoms further. Additionally, we examine the multifaceted management strategies for IBS, emphasizing the potential therapeutic benefits of dietary polyphenols as antioxidants. The present study aims to enhance our understanding of IBS and offer insights into more effective treatment strategies for this challenging condition.
Utilising Hyperspectral Autofluorescence Imaging in the Objective Assessment of Disease State and Pain in Patients with Rheumatoid Arthritis
Authors: Lees Florence , Mahbub Saabah B. , Gosnell Martin E. , Campbell Jared M. , Weedon Helen , Habibalahi Abbas , Goldys Ewa M. , Wechalekar Mihir D. , Hutchinson Mark R. , Crotti Tania N.
Published: November 2024

Rheumatoid Arthritis (RA) is a chronic inflammatory disease resulting in joint swelling and pain. Treatment options can be reliant on disease activity scores (DAS) incorporating patient global assessments, which are quantified via visual analogue scales (VAS). VAS can be subjective and not necessarily align with clinical symptoms, such as inflammation, resulting in a disconnect between the patient’s and practitioners’ experience. The development of more objective assessments of pain would enable a more targeted and personalised management of pain within individuals with RA and have the potential to improve the reliability of assessments in research. Using emerging light-based hyperspectral autofluorescence imaging (HAI) technology, we aimed to objectively differentiate disease and pain states based on the analysis of synovial tissue (ST) samples from RA patients. In total, 22 individuals with RA were dichotomised using the DAS in 28-joint counts (DAS-28) into an inactive (IA) or active disease (active-RA) group and then three sub-levels of pain (low, mid, high) based on VAS. HAI was performed on ST sections to identify and quantify the most prominent fluorophores. HAI fluorophore analysis revealed a distinct separation between the IA-RA and active-RA mid-VAS cohort, successfully determining disease state. Additionally, the separation between active-RA Mid-VAS and active RA High-VAS cohort suggests that HAI could be used to objectively separate individuals based on pain severity.
Sensitive Detection of Gynecological Cancer Recurrence Using Circulating Tumor DNA and Digital PCR: A Comparative Study with Serum Biochemical Markers
Authors: Balasan Nour , Kharrat Feras , Di Lorenzo Giovanni , Athanasakis Emmanouil , Bianco Anna Monica , Conti Andrea , Di Stazio Maria Teresa , Butera Giulia , Cicogna Stefania , Mangogna Alessandro , Romano Federico , Ricci Giuseppe , d’Adamo Adamo Pio
Published: November 2024

Early detection of recurrences in gynecological cancers is crucial for women’s health. Circulating tumor DNA (ctDNA) analysis through liquid biopsy offers a promising approach for monitoring disease progression and identifying relapses. This study investigated the utility of digital Polymerase Chain Reaction (dPCR) for ctDNA detection in three gynecological cancer patients with clinically confirmed relapses during a two-year post-surgical follow-up. Patient-specific tumor mutations were identified through whole-exome sequencing (WES) and confirmed via Sanger sequencing. dPCR probes targeting these mutations were used to quantify the ctDNA levels in plasma samples collected throughout the follow-up period, and the findings were compared with standard serum biochemical markers. In two patients, persistent positive dPCR signals for the selected mutations were detected after tumor removal, with ctDNA levels progressively increasing even after post-surgical chemotherapy. Notably, dPCR identified elevated ctDNA levels before an increase in the cancer antigen 125 (CA125) biochemical marker was observed. In the third patient, no ctDNA signals from the two selected mutations were detected despite clinical evidence of recurrence, suggesting the emergence of new mutations. While this study highlights the promise of dPCR for early recurrence detection in gynecological cancers, it also underscores the critical need for comprehensive mutation panels to overcome the inherent challenges posed by tumor heterogeneity and the emergence of new mutations during disease progression.
The Identification of a Novel Pathogenic Variant in the GATA6 Gene in a Child with Neonatal Diabetes
Authors: Sechko Elena A. , Koltakova Maria P. , Khusainova Rita I. , Minniakhmetov Ildar R. , Laptev Dmitry N.
Published: November 2024

GATA6 syndrome is a rare monogenic disorder caused by heterozygous variants in the gene GATA6, which controls the early embryonic differentiation of germ layers and the development of different organs. We present the results of the 7-year follow-up of a child with this syndrome as well as the following conditions: diabetes mellitus, exocrine pancreatic insufficiency, gallbladder atresia, and congenital heart disease (CHD). At birth, the patient was diagnosed with neonatal diabetes mellitus (NDM) associated with heart (mitral valve prolapse) and gastrointestinal abnormalities (gallbladder atresia). Diabetes remitted within weeks and relapsed at the age of 2. We identified a de novo variant of a 4-nucleotide deletion (c.1302+4_1302+7del), previously unreported in the literature, in the donor splicing site of exon 3 of the GATA6 gene in a heterozygous state. Screening for other possible components of GATA6 syndrome revealed exocrine pancreatic insufficiency, and pancreatic enzyme replacement therapy resulted in improved dyspeptic symptoms, and growth rates increased. In addition, the patient was diagnosed with autoimmune thyroiditis and progressive myopia.
Novel Allelic Mutations in Dw3 Gene That Affect the Height of Sorghum Plants
Authors: Wang Ping , Liang Bingbing , Li Zhengjun , Wang Chunyu , Zhang Lixia , Lu Xiaochun
Published: November 2024

Breeding for dwarfing traits in sorghum is crucial. However, only three genes (Dw1–Dw3) that control plant height have been mapped. In this study, 634 sorghum cultivars were collected to investigate plant height and genotypes. Four were genotyped Dw1DW2Dw3 (wild type) but with different plant heights, and they were selected to construct two populations and map new dwarf genes. Bulked segregant analysis with whole-genome resequencing of the two populations identified the candidate gene in one same genomic region—on chromosome 7. Then, it was narrowed down to a region containing nine genes. Amino acid and DNA sequence analysis of the parent and offspring plants revealed that two novel allelic mutations in the Dw3 gene play a role in reducing the plant height—8R262 or 8R417, including 1 bp substitution and 2 bp deletions. Furthermore, we sequenced 19 cultivars that primarily exhibited a “one-dwarf” hybrid or wild-type and presumed another allelic mutation via the amino acid alignment of 8R019, 8R100, and 8R402, which was another one-base substitution. These results indicate that multiple types of allelic mutations in the Dw3 gene should be considered when identified or applied.
Liver Fluke-Derived Molecules Accelerate Skin Repair Processes in a Mouse Model of Type 2 Diabetes Mellitus
Authors: Kovner Anna , Kapushchak Yaroslav , Zaparina Oxana , Ponomarev Dmitry , Pakharukova Maria
Published: November 2024

Chronic nonhealing wounds, such as diabetic ulcers, are among the most serious complications of diabetes mellitus. Consequently, the search for new therapeutic strategies remains highly relevant. Based on our previous data on acute wounds, bioactive molecules derived from the liver fluke Opisthorchis felineus hold promise as a novel approach to wound healing. The aim of this study was to investigate the wound-healing properties of excretory–secretory products (ESP) and inactivated eggs of O. felineus in a model of type 2 diabetes mellitus. Two-month-old mice of the BKS.Cg + Leprdb/+Leprdb/OlaHsd (db/db) strain were inflicted with superficial wounds of 5 mm in diameter. Mouse groups included several controls (methylcellulose as the vehicle and human recombinant PDGF as the positive control) and specific-treatment groups (ESP and inactivated O. felineus eggs). Histopathological, immunohistochemical, and RT-PCR studies using markers for M1/M2 polarization, angiogenesis, and extracellular matrix remodeling were carried out. Additionally, an image analysis of Masson’s trichrome-stained skin sections was performed. The proliferation of HaCaT cells under ESP and egg treatment was also assessed. The present study reveals a significant increase in the percentage of wound healing in ESP- and egg-treated groups, which significantly exceeded the control values after 14 days. Wound treatment with either ESP or worm eggs resulted in (i) a reduction in inflammation with a canonical M1-to-M2 polarization shift, (ii) the modulation of the vascular response, and (iii) dermal extracellular matrix remodeling. All results are comparable to those of the positive control group treated with PDGF. This study also reveals that ESP, but not O. felineus eggs, stimulated keratinocyte proliferation in vitro. The results indicate the high wound-healing potential of liver fluke bioactive molecules and open prospects for further research on these new promising therapeutic approaches.
Caffeine: A Multifunctional Efficacious Molecule with Diverse Health Implications and Emerging Delivery Systems
Authors: Song Xinjie , Singh Mahendra , Lee Kyung Eun , Vinayagam Ramachandran , Kang Sang Gu
Published: November 2024

Natural caffeine is found in many plants, including coffee beans, cacao beans, and tea leaves. Around the world, many beverages, including coffee, tea, energy drinks, and some soft drinks, have this natural caffeine compound. This paper reviewed the results of meta-studies on caffeine’s effects on chronic diseases. Of importance, many meta-studies have shown that regularly drinking caffeine or caffeinated coffee significantly reduces the risk of developing Alzheimer’s disease, epilepsy, and Parkinson’s disease. Based on the health supplements of caffeine, this review summarizes various aspects related to the application of caffeine, including its pharmacokinetics, and various functional health benefits of caffeine, such as its effects on the central nervous system. The importance of caffeine and its use in alleviating or treating cancer, diabetes, eye diseases, autoimmune diseases, and cardiovascular diseases is also discussed. Overall, consuming caffeine daily in drinks containing antioxidant and neuroprotective properties, such as coffee, prevents progressive neurodegenerative diseases, such as Alzheimer’s and Parkinson’s. Furthermore, to effectively deliver caffeine to the body, recently developed nanoformulations using caffeine, for instance, nanoparticles, liposomes, etc., are summarized along with regulatory and safety considerations for caffeine. The U.S. Department of Agriculture (USDA) and the Food and Drug Administration (FDA) recommended that healthy adults consume up to 400 mg of caffeine per day or 5~6 mg/kg body weight. Since a cup of coffee contains, on average, 100 to 150 mg of coffee, 1 to 3 cups of coffee may help prevent chronic diseases. Furthermore, this review summarizes various interesting and important areas of research on caffeine and its applications related to human health.
Sex Differences in the Neuroendocrine Stress Response: A View from a CRH-Reporting Mouse Line
Authors: Horváth Krisztina , Vági Pál , Juhász Balázs , Kuti Dániel , Ferenczi Szilamér , Kovács Krisztina J.
Published: November 2024

Corticotropin-releasing hormone (CRH) neurons within the paraventricular hypothalamic nucleus (PVH) play a crucial role in initiating the neuroendocrine response to stress and are also pivotal in coordination of autonomic, metabolic, and behavioral stress reactions. Although the role of parvocellular CRHPVH neurons in activation of the hypothalamic–pituitary–adrenal (HPA) axis is well established, the distribution and function of CRH-expressing neurons across the whole central nervous system are less understood. Stress responses activate complex neural networks, which differ depending on the type of stressor and on the sex of the individual. Because of the technical difficulties of localizing CRH neurons throughout the rodent brain, several CRH reporter mouse lines have recently been developed. In this study, we used Crh-IRES-Cre;Ai9 reporter mice to examine whether CRH neurons are recruited in a stressor- or sex-specific manner, both within and outside the hypothalamus. In contrast to the clear sexual dimorphism of CRH-mRNA-expressing neurons, quantification of CRH-reporting, tdTomato-positive neurons in different stress-related brain areas revealed only subtle differences between male and female subjects. These results strongly imply that sex differences in CRH mRNA expression occur later in development under the influence of sex steroids and reflects the limitations of using genetic reporter constructs to reveal the current physiological/transcriptional status of a specific neuron population. Next, we compared the recruitment of stress-related, tdTomato-expressing (putative CRH) neurons in male and female Crh-IRES-Cre;Ai9 reporter mice that had been exposed to predator odor. In male mice, fox odor triggered more c-Fos in the CRH neurons of the paraventricular hypothalamic nucleus, central amygdala, and anterolateral bed nucleus of the stria terminalis compared to females. These results indicate that male mice are more sensitive to predator exposure due to a combination of hormonal, environmental, and behavioral factors.
The IL-33/ST2 Axis Affects Adipogenesis Through Regulating the TRAF6/RelA Pathway
Authors: Cao Shujun , Qin Xuyong , Li Chengping , Zhang Lichun , Ren Shizhong , Zhou Wenhao , Zhao Meiman , Zhou Guoli
Published: November 2024

Understanding the regulatory mechanisms of adipogenesis is essential for preventing obesity. Interleukin-33 (IL-33) has recently attracted increasing attention for its role in adipogenesis. The purpose of this study was to explore the function and regulatory mechanism of IL-33 and its receptor suppression of tumorigenicity 2 (ST2) on adipogenesis. Here, Oil Red O staining was used to detect the accumulation of intracellular lipid droplets. Molecular techniques such as qRT-PCR and Western blotting were used to detect the expression of pivotal genes and adipogenic marker genes. Gains and losses of function experiments were used to explore the potential regulatory mechanism of the IL-33/ST2 axis in adipogenesis. Functionally, IL-33 is negatively associated with adipogenesis in 3T3-L1 preadipocytes, while ST2 is positively associated with it, encompassing both the trans-membrane receptor ST2 (ST2L) and the soluble ST2 (sST2). Mechanistically, the IL-33/ST2 axis affects adipogenesis by regulating the expression of the TRAF6/RelA pathway in 3T3-L1 preadipocytes. Downregulating the expression of ST2 suppressed the activation of the IL-33/ST2 axis, which subsequently inhibits the expression of TRAF6. This further attenuates the expression of RelA, ultimately resulting in the suppression of adipogenesis in 3T3-L1 preadipocytes. This study reveals a new mechanism by which the IL-33/ST2 axis regulates the differentiation of preadipocytes and provides a new idea for improving obesity prevention.
Effect of Diosgenin in Suppressing Viability and Promoting Apoptosis of Human Prostate Cancer Cells: An Interplay with the G Protein-Coupled Oestrogen Receptor?
Authors: Figueira Marília I. , Marques Ricardo , Cardoso Henrique J. , Fonseca Lara R. S. , Duarte Ana P. , Silvestre Samuel , Socorro Sílvia
Published: November 2024

Diosgenin is a phytosteroid sapogenin with reported antitumoral activity. Despite the evidence indicating a lower incidence of prostate cancer (PCa) associated with a higher consumption of phytosteroids and the beneficial role of these compounds, only a few studies have investigated the effects of diosgenin in PCa, and its mechanisms of action remain to be disclosed. The present study investigated the effect of diosgenin in modulating PCa cell fate and glycolytic metabolism and explored its potential interplay with G protein-coupled oestrogen receptor (GPER). Non-neoplastic (PNT1A) and neoplastic (LNCaP, DU145, and PC3) human prostate cell lines were stimulated with diosgenin in the presence or absence of the GPER agonist G1 and upon GPER knockdown. Diosgenin decreased the cell viability, as indicated by the MTT assay results, which also demonstrated that castrate-resistant PCa cells were the most sensitive to treatment (PC3 > DU145 > LNCaP > PNT1A; IC50 values of 14.02, 23.21, 56.12, and 66.10 µM, respectively). Apoptosis was enhanced in diosgenin-treated cells, based on the increased caspase-3-like activity, underpinned by the altered expression of apoptosis regulators evaluated by Western blot analysis, which indicated the activation of the extrinsic pathway. Exposure to diosgenin also altered glucose metabolism. Overall, the effects of diosgenin were potentiated in the presence of G1. Moreover, diosgenin treatment augmented GPER expression, and the knockdown of the GPER gene suppressed the proapoptotic effects of diosgenin in PC3 cells. Our results support the antitumorigenic role of diosgenin and its interest in PCa therapy, alone or in combination with G1, mainly targeting the more aggressive stages of the disease.
Role of Peptide Associations in Enhancing the Antimicrobial Activity of Adepantins: Comparative Molecular Dynamics Simulations and Design Assessments
Authors: Maleš Matko , Juretić Davor , Zoranić Larisa
Published: November 2024

Adepantins are peptides designed to optimize antimicrobial biological activity through the choice of specific amino acid residues, resulting in helical and amphipathic structures. This paper focuses on revealing the atomistic details of the mechanism of action of Adepantins and aligning design concepts with peptide behavior through simulation results. Notably, Adepantin-1a exhibits a broad spectrum of activity against both Gram-positive and Gram-negative bacteria, while Adepantin-1 has a narrow spectrum of activity against Gram-negative bacteria. The simulation results showed that one of the main differences is the extent of aggregation. Both peptides exhibit a strong tendency to cluster due to the amphipathicity embedded during design process. However, the more potent Adepantin-1a forms smaller aggregates than Adepantin-1, confirming the idea that the optimal aggregations, not the strongest aggregations, favor activity. Additionally, we show that incorporation of the cell penetration region affects the mechanisms of action of Adepantin-1a and promotes stronger binding to anionic and neutral membranes.
RETRACTED: Al-Wahaibi et al. Optimized Conjugation of Fluvastatin to HIV-1 TAT Displays Enhanced Pro-Apoptotic Activity in HepG2 Cells. Int. J. Mol. Sci. 2020, 21, 4138
Authors: Al-Wahaibi Lamya H. , Al-Saleem Muneera S. M. , Ahmed Osama A. A. , Fahmy Usama A. , Alhakamy Nabil A. , Eid Basma G. , Abdel-Naim Ashraf B. , Abdel-Mageed Wael M. , AlRasheed Maha M. , Shazly Gamal A.
Published: November 2024

The journal retracts the article titled “Optimized Conjugation of Fluvastatin to HIV-1 TAT Displays Enhanced Pro-Apoptotic Activity in HepG2 Cells” [...]
Encapsulating Proton Inside C60 Fullerene: A Density Functional Theory Study on the Electronic Properties of Cationic X+@C60 (X+ = H+, H3O+ and NH4+)
Authors: Zhao Lei , Wang Bo
Published: November 2024

Confining protons into an enclosed carbon cage is expected to give rise to unique electronic properties for both the inner proton and the outer cage. In this work, we systematically investigated the geometric and electronic structures of cationic X+@C60 (X+ = H+, H3O+, and NH4+), and their corresponding neutral species (X = H2O, NH3), by quantum chemical density functional theory calculations. We show that C60 can trap H2O, NH3, H3O+ and NH4+ at the cage center and only slightly influence their geometries. The single proton clings to the inner wall of C60, forming a C-H chemical bond. The encapsulated neutral species almost do not change the electronic structure of the C60, while the internal cations have obvious effects. The charge transfer effect from the inner species to the C60 cage was found for all X@C60 (X = H2O, NH3) (about 0.0 e), X+@C60 (X+ = H3O+, NH4+) (about 0.5 e) and H+@C60 (about 1.0 e) systems. Encapsulating different forms of protons also regulates the fundamental physico-chemical properties of the hollow C60, such as the HOMO-LUMO gaps, infrared spectra, and electrostatic potential, etc., which are discussed in detail. These findings provide a theoretical insight into protons’ applications, especially in energy.
Accumulation of Bioactive Lipid Species in LPS-Induced Neuroinflammation Models Analysed with Multi-Modal Mass Spectrometry Imaging
Authors: Razo Irma Berrueta , Shea Kerry , Allen Tiffany-Jayne , Boutin Hervé , McMahon Adam , Lockyer Nicholas , Hart Philippa J.
Published: November 2024

Neuroinflammation is a complex biological process related to a variety of pathologies, often requiring better understanding in order to develop new, targeted therapeutic interventions. Within this context, multimodal Mass Spectrometry Imaging (MSI) has been used to characterise molecular changes in neuroinflammation for biomarker discovery not possible to other techniques. In this study, molecules including bioactive lipids were detected across inflamed regions of the brain in rats treated with lipopolysaccharide (LPS). The detected lipids may be acting as inflammatory mediators of the immune response. We identified that N-acyl-phosphatidylethanolamine (NAPE) species accumulated in the inflamed area. The presence of these lipids could be related to the endocannabinoid (eCB) signalling system, mediating an anti-inflammatory response from microglial cells at the site of injury to balance pro-inflammation and support neuronal protection. In addition, polyunsaturated fatty acids (PUFAs), specifically n-3 and n-6 species, were observed to accumulate in the area where LPS was injected. PUFAs are directly linked to anti-inflammatory mediators resolving inflammation. Finally, acylcarnitine species accumulated around the inflammation region. Accumulation of these molecules could be due to a deficient β-oxidation cycle.
Associations of Left Atrial Volume Index to Left Ventricular Ejection Fraction Ratio with Clinical Outcomes in Transthyretin Cardiac Amyloidosis
Authors: Aleligne Yeabsra K. , Wilson Machelle D. , Cadeiras Martin , Gibson Michael , Jimenez Shirin , Yala Stella , Acevedo Pablo E. , Liem David A. , Bidwell Julie T. , Ebong Imo A.
Published: November 2024

Background: Transthyretin amyloid cardiomyopathy (ATTR-CM) affects all cardiac chambers to cause left ventricular (LV) deformation as well as left atrial (LA) remodeling and functional impairment. We investigated the associations of the LA volume index (LAVI):LV ejection fraction (LVEF) ratio with the increased risk of death, heart transplant, or LV assist device implantation (LVAD) in patients with ATTR-CM. Methods: This was a retrospective cohort study involving 69 heart failure (HF) patients with ATTR-CM at an academic medical center between 1 November 2008 and 31 March 2024. ATTR-CM was diagnosed using a technetium–diphosphonate/pyrophosphate scan or an endomyocardial biopsy. The LAVI and LVEF were measured by echocardiography. Cox proportional hazards models were used for the analysis. Results: The mean (SD) age of the participants was 77.5 (9.3) years. Over a median (IQR) follow-up period of 1.96 (0.67–2.82) years, we observed 24 composite events that included twenty-two deaths, two heart transplants, and two LVAD implantations (who subsequently died). In multivariable-adjusted analyses that accounted for age and the glomerular filtration rate, a one-unit increase in the LAVI:LVEF ratio was associated with a doubling of the risk (HR, 95% CI: 2.06, 1.11–3.82) of experiencing the composite outcome. Conclusions: A one-unit increase in the LAVI:LVEF ratio was associated with an increased risk of death, heart transplant, or LVAD implantation in patients with ATTR-CM.
Tricuspid Valve Reconstruction in Patients with Right Heart Decompensation Due to Severe Tricuspid Regurgitation on LVAD Support
Authors: Welp Henryk , Sindermann Jürgen , Scherer Mirela
Published: November 2024

Background/Objectives: Right ventricular (RV) dysfunction after left ventricular assist device (LVAD) implantation is associated with reduced survival and affects duration of hospitalization. Some patients with RV dysfunction on LVAD have significant tricuspid valve regurgitation (TR) with concomitant signs of RV failure. In these cases, tricuspid valve repair (TVR) may minimize clinical signs of RV failure. Methods: We report on two patients (one female, one male) developing significant TR receiving TVR through a right thoracotomy on LVAD support. Results: The time between LVAD implantation and TVR was 4 months and 50 months, respectively. The female patient could be discharged from hospital without TR and any signs of RV dysfunction. The male patient died 13 days later due to septic shock following mesenteric ischemia. At this time, echocardiography showed a competent tricuspid valve. Conclusions: In conclusion, these results provide insight into the clinical judgment of when TVR should be attempted. They suggest whether TVR may be a strategy to avoid hospitalization, minimize the clinical signs of RV insufficiency and improve quality of life in patients on LVAD support with severe TR and clinical signs of right heart dysfunction.
The Efficacy of Andexanet Alfa for the Reversal of Factor Xa Inhibitors Is Not Influenced by Hemodilution with Different Volume Expanders
Authors: Wienhold Jan , Rossaint Rolf , Vandeput Eline , Grottke Oliver
Published: November 2024

Background: Andexanet alfa is a specific antidote for factor Xa (FXa) inhibitors. It is licensed to treat patients under FXa inhibitor therapy with life-threatening bleeding. Concomitantly, volume expanders are used to compensate for blood loss and maintain circulation. The competitive binding of andexanet to FXa inhibitors may be disrupted due to hemodilution, as shown by laboratory assays with high sample dilution. This study investigated the efficacy of andexanet for the reversal of FXa inhibitors under hemodilution. Methods: Blood from 10 healthy volunteers was anticoagulated with rivaroxaban and subsequently treated with four different volume expanders (Ringer’s solution, 4% gelatine, 5% and 20% human albumin (HA)) at two dilution levels (20% and 50%). After anticoagulation and hemodilution, andexanet was added according to the high-dose protocol. Blood samples were analyzed using a Russell’s viper venom (RVV) test on a Clot Pro® device, a thrombin generation assay, a fully automated coagulation analyzer and a chromogenic anti-FXa activity assay. Results: After anticoagulation, the median rivaroxaban concentration was 272 ng/mL (IQR 254–353). Anticoagulation with rivaroxaban caused a significant impairment of all coagulation parameters, which was further aggravated by hemodilution. After the administration of andexanet, coagulation parameters in anticoagulated samples were reversed to near baseline in all groups. Andexanet administration decreased the rivaroxaban plasma concentration in all groups to a median of <10 ng/mL. In the anticoagulated, non-hemodiluted samples, anti-FXa activity was reduced by 98%. The anti-FXa activity in the anticoagulated, hemodiluted samples was reduced by approximately 96% in the 20% diluted samples and by about 93% in the 50% diluted samples. Conclusions: Our data indicate that FXa inhibitor reversal with andexanet is about 5% less effective with 50% hemodilution than in non-hemodiluted samples.