bluuebunny/crossref_metadata_2025 huggingface dataset:
Highlighting the Importance of Correct Sex Identification in Chondrichthyan Genomic Studies, Using the White Shark as an Example
Authors: Lee Szu-Hsuan , Yang Lei , Naylor Gavin J. P.
Published: December 2024

The increased availability of reference genome assemblies of sharks and rays has contributed greatly to our understanding of their biology, including their sex-determination mechanisms. However, several publicly available genome assemblies of sharks and rays appear to be missing information about the sex of the source individuals. This can confound the investigation into genetic sex-determining elements and hinder the discovery of sex-specific patterns. Herein, we highlight the importance of clear and accurate sex identification in sharks and rays for future genome assemblies, using an example of a white shark (Carcharodon carcharias) genome, in which the phenotypically assigned sex conflicts with the genetic information. This genome assembly was reported to be sourced from a juvenile female (BioSample: SAMN01915239). We analyzed the assembly by mapping its available genome sequences to the current white shark reference genome assembly and compared the read coverage to sequences collected from other samples. Evidence suggests that this specimen is genetically male, which contradicts its assignment based on phenotype. Therefore, we urge researchers to provide as much accurate information (e.g., sex, sampling localities, and life history) as possible when publishing genome assemblies for sharks and rays (or for any other organism).
Nutrient Stability in NASA Spaceflight Experiment Rodent Food Bars
Authors: Iyer Janani , Marsh Tyler S , Fisher Ryan J , Verma Vandana
Published: December 2024

The Nutrient-upgraded Rodent Food Bar (NuRFB) is the standard diet for mice in NASA’s Rodent Research Project aboard the International Space Station (ISS). Given the nature of spaceflight and the lengthy production process of the food bars, a shelf-life assessment was conducted to evaluate nutritional stability over time (ranging from 0 to 27 months) and under different storage conditions (refrigerated, ambient, and refrigerated + ambient), where ambient is 22–23 °C. Lipid oxidation markers and fat- and water-soluble vitamins were assessed under various time and temperature conditions using AOAC International methods. Vitamin D levels showed a minor decrease, and riboflavin fluctuated slightly over time, but all vitamin levels remained above National Research Council (NRC) minimum requirements. Food bars stored at 4 °C showed significantly higher thiamine levels than the bars that underwent some degree of ambient temperature storage, but all met the NRC guidelines. Minimal lipid oxidation was observed for up to 18 months, and no mold or yeast growth occurred despite the high moisture content of the bars. This study confirms that NuRFBs maintain stable vitamin and lipid oxidation indices, ensuring adequate nutrition for rodents during spaceflight.
In Vitro Evaluation of Probiotic Activities and Anti-Obesity Effects of Enterococcus faecalis EF-1 in Mice Fed a High-Fat Diet
Authors: Cai Hongying , Wang Qingya , Han Xiling , Zhang Haiou , Wang Na , Huang Yuyin , Yang Peilong , Zhang Rui , Meng Kun
Published: December 2024

This research sought to assess the anti-obesity potential of Enterococcus faecalis EF-1. An extensive and robust in vitro methodology confirmed EF-1’s significant potential in combating obesity, probably due to its excellent gastrointestinal tract adaptability, cholesterol-lowering property, bile salt hydrolase activity, α-glucosidase inhibition, and fatty acid absorption ability. Moreover, EF-1 exhibited antimicrobial activity against several pathogenic strains, lacked hemolytic activity, and was sensitive to all antibiotics tested. To further investigate EF-1’s anti-obesity properties in vivo, a high-fat diet (HFD) was used to induce obesity in C57BL/6J mice. Treatment with EF-1 (2 × 109 CFU/day) mitigated HFD-induced body weight gain, reduced adipose tissue weight, and preserved liver function. EF-1 also ameliorated obesity-associated microbiota imbalances, such as decreasing the Firmicutes/Bacteroidetes ratio and boosting the levels of bacteria (Faecalibacterium, Mucispirillum, Desulfovibrio, Bacteroides, and Lachnospiraceae_NK4A136_group), which are responsible for the generation of short-chain fatty acids (SCFAs). Concurrently, the levels of total SCFAs were elevated. Thus, following comprehensive safety and efficacy assessments in vitro and in vivo, our results demonstrate that E. faecalis EF-1 inhibits HFD-induced obesity through the regulation of gut microbiota and enhancing SCFA production. This strain appears to be a highly promising candidate for anti-obesity therapeutics or functional foods.
Effect of Different Formulations and Storage on the Physicochemical, Microbiological, and Organoleptic Characteristics of Dovyalis caffra Fruit Yogurt
Authors: Waweru Daniel Mwangi , Arimi Joshua Mbaabu , Marete Eunice , Harbourne Niamh , Jacquier Jean-Christophe
Published: December 2024

This study investigated the effect of incorporating up to 15% (w/w) Dovyalis caffra fruit pulp into cow milk yogurt. Monitoring the physico-chemical, microbiological, and organoleptic properties of these formulations was performed weekly during refrigerated (4 °C) storage for 21 days. Compared to the control, formulations with added pulp recorded enhanced contents of ascorbic acid, total polyphenols, fiber, total titratable acidity (TTA), and yellowness, which is in line with increasing fruit pulp. This, however, corresponded to significantly lower contents in fat, protein, viscosity, pH, and overall sensory acceptability. Pulp incorporation had no significant effect on moisture, ash, and water-holding capacity (WHC) of formulations. Storage for up to 21 days indicated a significant increase in TTA and a corresponding decrease in pH but no significant change in WHC and viscosity of formulations. There was no detection of coliforms, yeasts, and molds in all samples throughout the storage period. This study demonstrates that Dovyalis caffra fruit has significant potential in the formulation of nutritious, desirable, and shelf-stable fruit-based yogurts. Further product optimization is, however, recommended to maximize the organoleptic quality of the formulations.
Influence of Seed Disinfection Treatments on the Germination Rate and Histamine-Degrading Activity of Legume Sprouts
Authors: Costa-Catala Judit , Bori Jaume , Veciana-Nogués M. Teresa , Latorre-Moratalla M. Luz , Vidal-Carou M. Carmen , Comas-Basté Oriol
Published: December 2024

Edible legume sprouts have been proposed as a promising plant-based source of the enzyme diamine oxidase (DAO), which plays a key role in degrading histamine at an intestinal level and preventing the development of histamine intolerance symptoms. However, the temperature and humidity conditions required for seed germination can also favor the rapid growth of yeast and mold, potentially compromising sprout yield and quality. The aim of this study was to evaluate the influence of different seed disinfection treatments on both the germination rate and DAO enzymatic activity in sprouts of four Leguminosae species. Seed disinfection with 70% ethanol for either 5 or 15 min slightly increased the germination rates of chickpea and soybean sprouts without affecting DAO activity, regardless of treatment duration. However, in lentil and green pea sprouts, ethanol disinfection caused a statistically significant reduction in histamine-degrading capacity. In contrast, treating seeds with sodium hypochlorite for 15 min increased germination rates by up to 14% and preserved DAO activity in all legume sprouts tested. These results indicate that incorporating a seed disinfection step during legume sprouting may affect both the DAO enzymatic activity and germination rate.
The Complete Mitochondrial Genome of the Luciocyprinus langsoni (Cypriniformes: Cyprinidae): Characterization, Phylogeny, and Genetic Diversity Analysis
Authors: Yang Tiezhu , Tan Chenxi , Zhao Liangjie , Hu Zhiguo , Su Chaoqun , Li Fan , Ma Yuanye , Zhang Wenchao , Hao Xiaoyu , Zou Wenxu , Kang Jiayin , He Qingqing
Published: December 2024

Background: Luciocyprinus langsoni is a species belonging to the Cyprinidae family. The objective of this study is to gain a comprehensive understanding of its evolutionary history and genetic characteristics. Methods: The complete mitochondrial genome of L. langsoni was determined using overlapping PCR. A phylogenetic analysis was conducted based on 13 protein-coding genes from 48 species. A population genetic diversity analysis using the COI gene and a selection analysis of 13 protein-coding genes were also performed. Results: The mitogenome is 16,586 base pairs long and consists of 13 protein-coding genes, two ribosomal RNAs, 22 transfer RNAs, and two control regions. It has a high adenine-thymine (A + T) content. The phylogenetic analysis confirms the placement of L. langsoni within the subfamily Cyprininae. The population genetic diversity analysis reveals low variability in the Hechi Longjiang population. The selection analysis shows that all 13 protein-coding genes have evolved under purifying selection with Ka/Ks ratios below 1. Conclusions: These results enhance our understanding of L. langsoni’s evolutionary history and lay a genetic foundation for future studies in population genetics and phylogenetics.
Evaluating the Impact of Geoeducation Programs on Student Learning and Geoheritage Awareness in Greece
Authors: Zafeiropoulos George , Drinia Hara
Published: December 2024

In recent years, multiple efforts to promote geoscience and geoenvironmental awareness have highlighted the need for geoeducation. This article explores the importance of integrating geoeducation into the Greek curriculum for promoting a deeper understanding of geosciences. The method used is primarily based on a mixed-methods approach, covering both qualitative and quantitative data. A preliminary assessment revealed a lack of information on the conceptual framework of geosites and geoparks. Following this, a geoeducation tool was developed to introduce the basic principles of geoeducation. The evaluation showed a significant increase in understanding and a heightened desire for further knowledge on geoeducational topics. To address current deficiencies, effective integration of this geoeducational tool requires comprehensive teacher training and curriculum reform. Promoting geoeducation through school programs is critical for creating a more informed, responsible, and sustainable society. This paper advocates policy changes and educational initiatives to firmly integrate geoeducation into the Greek educational system.
Characterisation of Pelletal Lapilli in Explosive Melilitite–Carbonatite Eruptions: An Example from Mt. Vulture Volcano (Southern Italy)
Authors: Carnevale Gabriele , Zanon Vittorio
Published: December 2024

Among the volcaniclastic products of melilitite–carbonatite eruptions, pelletal lapilli are often found, resulting in them being particularly useful for characterising the interface between the erupting magma and its volatile component. Pelletal lapilli, which were erupted during the most recent melilitite–carbonatite volcanic activity of the Mt. Vulture volcano, are characterised by a predominantly wehrlitic core with CO2-rich fluid inclusions and a Ca-rich outer portion composed of fine-grained xenocrystic debris of olivine and clinopyroxene, with microcrysts of haüyne and melilite laths (± calcite). The chemical composition of the olivine reflects the interaction with a proto-melilitite–carbonatite melt, which is the main metasomatic agent. The whole-rock analyses of the external portion of pelletal lapilli show values that are comparable with those of extrusive carbonatites. This evidence supports the hypothesis that the primary carbonatite melt was a significant contributor to the CO2-rich magma source that transported the lapilli to the surface. The modelling of the geometric data of the pelletal lapilli structure, together with inferences regarding the role of the CO2 gas phase, the main propellant in an ascending gas-dominated medium, allowed for the reconstruction of a possible scenario where the CO2 expansion and the fluidised spray granulation process are crucial during the volcanic conduit dynamics.
The Effect of Immediate Kangaroo Mother Care During Third Stage of Labor on Postpartum Blood Loss and Uterine Involution: A Quasi-Experimental Comparative Study
Authors: Almutairi Wedad M. , Raidi Dareen K.
Published: December 2024

Background: Obstetric hemorrhage is the leading cause of maternal death worldwide. Obstetric hemorrhage accounts for 27.1% of all maternal death worldwide. Of all obstetric hemorrhages, postpartum hemorrhage (PPH) accounts for 72%. The physiological management of the third stage of labor is a growing area as a preventive measure to control postpartum blood loss. Immediate kangaroo mother care (KMC) is suggested as one of the physiological management methods of the third stage of labor to reduce postpartum blood loss. The duration of the third stage of labor, uterine involution, and amount of postpartum blood loss are the physiological parameters of effective management of the third stage of labor. Examining the absolute effects of immediate KMC on maternal physiological parameters is needed in different populations with different settings. Thus, this study aimed to examine the effects of immediate KMC on uterine involution and postpartum blood loss. Methods: A quasi-experimental comparative design was conducted in the labor and delivery room at Maternity and Children Hospital, Makkah, Saudi Arabia. A sample of 80 women was divided into two equal groups: a treatment group that underwent immediate KMC and a control group that received routine care. Instrument: A questionnaire developed by the researchers was used to collect the data. Results: The effects of immediate KMC were significant concerning uterine involution and regarding the uterine position immediately after placenta separation (70% at umbilicus, χ2 = 8.5, p < 0.01), postpartum blood loss (χ2 = 76.098, p < 0.00), the heaviness of lochia (χ2 = 44.679, p = 0.00), and the number of pads used in the first 24 h (p < 0.001).
User-Centred Design and Development of a Smartphone Application (OverSight) for Digital Phenotyping in Ophthalmology
Authors: Devraj Kishan , Jones Lee , Higgins Bethany , Thomas Peter B. M. , Moosajee Mariya
Published: December 2024

Background: Visual impairment can significantly impact an individual’s daily activities. Patients require regular monitoring, typically occurring within hospital eye services. Capacity constraints have necessitated innovative solutions to improve patient care. Existing digital solutions rely on task-based digital home monitoring such as visual acuity testing. These require active involvement from patients and do not typically offer an indication of quality of life. Digital phenotyping refers to the use of personal digital devices to quantify passive behaviour for detecting clinically significant changes in vision and act as biomarkers for disease. Its uniqueness lies in the ability to detect changes passively. The objective was to co-design an accessible smartphone app (OverSight) for the purposes of digital phenotyping in people with sight impairment. Methods: Development of OverSight included stakeholder consultations following principles of user-centred design. Apple iOS software frameworks (HealthKit, ResearchKit, and SensorKit) and a SwiftUI developer toolkit were used to enable the collection of active and passive data streams. Accessibility and usability were assessed using the System Usability Scale (SUS) and feedback following a 3-month pilot study. Consultations with patients informed the design of OverSight, including preferred survey scheduling and the relevancy of patient support resources. Results: Twenty visually impaired participants (mean age 42 ± 19 years) were recruited to the pilot study. The average score on the SUS was 76.8 (±8.9), indicating good usability. There was a statistically significant moderate negative correlation between SUS scores and visual acuity in both the better (r = −0.494; p ≤ 0.001) and worse eye (r = −0.421; p ≤ 0.001). Conclusions: OverSight offers promising potential for collecting patient-generated health data for the purposes of digital phenotyping in patients with eye disease. Through further testing and validation, this novel approach to patient care may ultimately provide opportunities for remote monitoring in ophthalmology.
Patient Engagement in Providing Telehealth SUD IOP Treatment: A Retrospective Cohort Study
Authors: Contreras-Schwartz Joanna , O’Neill Conor , Threlkeld Annelise , O’Callaghan Erin , Winsberg Mirene
Published: December 2024

Background: Substance use disorders (SUDs) remain a growing public health issue, with drug- and alcohol-related deaths continuing to increase. A myriad of barriers prevent many with SUDs from seeking care. Telehealth interventions are well-positioned to reduce barriers and increase engagement in SUD treatment. The SUD intensive outpatient program (IOP) is specifically designed for telehealth and offers evidenced-based care delivered by SUD professionals as well as asynchronous assignments to enhance treatment. This study explores the feasibility of providing a telehealth IOP. Methods: participant engagement, reasons for disengagement, and days of abstinence were examined using existing records from a cohort of participants between 2021 and 2023 (n = 4724). Results: Nearly 80% of participants remained engaged in the program for 30 days, and 91% attained at least 30 consecutive days of abstinence over the course of treatment. Nearly 45% demonstrated a successful response to care and no longer required IOP treatment. Those who finished the IOP completed over 70% of the asynchronous assignments. Conclusions: Results support the feasibility and effectiveness of delivering a telehealth IOP for SUDs.
Comprehensive Identification and Expression Profiling of Lipoxygenase Genes in Sweet Cherry During Fruit Development
Authors: Fu Quanjuan , Xu Di , Hou Sen , Gao Rui , Zhou Jie , Chen Chen , Zhu Shengnan , Wei Guoqin , Sun Yugang
Published: December 2024

Lipoxygenase (LOX) is involved in the oxidation of fatty acids in plants and is a ubiquitous oxygenase that plays an important role in the process of plant resistance to adversity. In this study, the LOX gene family in the sweet cherry genome was identified by bioinformatics methods, the chromosomal mapping of different LOX genes was carried out, and the homology alignment and functional domain analysis of the encoded proteins were performed. The results showed that there were nine LOX gene sequences in the sweet cherry LOX gene family, and the subcellular localization was mainly located in the cytoplasm, chloroplast, or plasma membrane, and was concentrated on chromosomes 1, 2, 3, 4, 6, and 8. During the ripening process of sweet cherry fruits, the LOX gene family showed five different expression patterns, the expression peak of different LOX genes reached the peak of expression at a specific development period, all LOX genes jointly promoted the growth and development of fruits, the enzyme activities of LOX in different varieties of early and late ripening cherries exhibited great differences during the development process, and the results of volatile content in the later stages also showed that different varieties of cherries had their specificity. The results of this study provide a theoretical basis for further revealing the specific functions of LOX gene family members in sweet cherry.
Main Barriers in Reducing Microbial Load in Raw Vegetables Served on Brazilian School Menus
Authors: Batista Sueny Andrade , dos Santos Emanuele Batistela , Câmara Gabriel Teles , Rose Ester Cardoso Paes , Dantas Pedro Tourinho , Botelho Raquel Braz Assunção , Ginani Verônica Cortez
Published: December 2024

This study assessed raw vegetable sanitizing in Brazilian schools and identified barriers to standards. This experimental and quantitative study was conducted in 12 school food services in the Federal District (Brazil) public primary education institutions. Microbiological analyses were conducted with vegetable samples (before and after sanitizing) and water used in the sanitization process, collected before the process. The Petrifilm®E. coli/Coliform Count Plates and COLIlert methods were used to evaluate vegetables and water samples, and a checklist of good practices was applied in each school food service to identify barriers to proper sanitization. Thirty-five samples of raw vegetables were offered to students, 32 samples of water, and 17 hygiene processes were evaluated. The results indicate that 76.5% (n = 13) of hygiene processes were considered unsatisfactory, with an average increase of 5.8 log CFU g−1 (DV = 7.4) in the initial microbial load in 47.1% (n = 8) of the evaluated processes; moreover, 33.3% (n = 6) of the samples exceeded the tolerable limit, with an average value above 1.5 × 103 CFU/g. Attention to food handler training and necessary organizational changes is essential to ensure safe food and promote healthy student eating habits, highlighting the importance of strengthening basic hygiene practices and following the parameters for sanitizing vegetables.
Cost Analysis Related to Diagnosis, Treatment and Management of Cervical Cancer in Antigua and Barbuda: A Prevalence-Based Cost-of-Illness Study
Authors: Bovell Andre A. N. , Ngcamphalala Cebisile , Abbott Dane , Ncayiyana Jabulani , Ginindza Themba G.
Published: December 2024

Cervical cancer remains a significant public health issue globally. In Antigua and Barbuda, cervical cancer is ranked among the top five most common cancers in terms of incidence and mortality among females. There is no evidence that the costs of diagnosing, treating, and managing this cancer have been studied before in Antigua and Barbuda. From the providers’ perspective, this study aimed to estimate costs associated with cervical cancer in Antigua and Barbuda. The prevalence-based cost-of-illness methodology was used to assess patient data abstracted from four study sites for the period 2017–2021, and to derive the annual prevalence. Top-down and bottom-up costing approaches were used to estimate direct medical costs. Costs were computed using the 2021 price level and converted to United States Dollars (USD). Total annual direct medical costs of cervical cancer were estimated at USD 0.24 million (ranging between USD 0.19 million and USD 0.30 million). Major cost drivers were treatment (USD 112,863.76), post-treatment side-effects care (USD 67,406.57), and the diagnostic process (USD 26,238.58). The overall direct medical unit costs for managing a case were estimated at USD 115,822.09. Our study reflects the current estimates for managing cervical cancer and provides evidence to complement cervical cancer prevention and cost containment measures in Antigua and Barbuda.
But at What Cost? Healthcare Utilization of Canadian Carer-Employees
Authors: Ding Regina , Duxbury Linda
Published: December 2024

Caregiving plays a crucial role in aging societies by supporting individuals with chronic illnesses, disabilities, or aging-related needs. The unpaid labour provided by caregivers diverts healthcare resources from the formal healthcare system; however, this incurs costs to the caregivers themselves in terms of declines in personal wellbeing. This study explores the relationship between caregiving and healthcare spending for two groups of caregivers: eldercare only and sandwiched. We found that physician visits were the most common resource used by caregivers, at a mean of 3.69 (SD = 4.01) visits over a 6-month period, excluding non-users of this service. This was followed by mental health services (M = 5.86, SD = 7.02), emergency room visits (M = 1.77, SD = 1.38), and hospital admissions (M = 3.61, SD = 8.53). There were no significant differences in healthcare utilization between eldercare and sandwiched caregivers. There were mixed results regarding characteristics associated with greater resource use; however, the hours of weekly caregiving were most consistently associated with greater healthcare utilization, indicating that healthcare use may increase with care burden.
Transcriptome and Gene Expression Analysis Revealed CeNA1: A Potential New Marker for Somatic Embryogenesis in Common Centaury (Centaurium erythraea Rafn.)
Authors: Ćuković Katarina B. , Todorović Slađana I. , Savić Jelena M. , Bogdanović Milica D.
Published: December 2024

Centaurium erythraea Rafn. is a medicinal plant used as a model for studying plant developmental processes due to its developmental plasticity and ease of manipulation in vitro. Identifying the genes involved in its organogenesis and somatic embryogenesis (SE) is the first step toward unraveling the molecular mechanisms underlying its morphogenic plasticity. Although SE is the most common method of centaury regeneration, the genes involved in this have not yet been identified. The aim of this study was to identify the differentially expressed genes (DEGs) during key stages of SE and organogenesis using transcriptome data, with a focus on novel SE-related genes. The transcriptomic analysis revealed a total of 4040 DEGs during SE and 12,708 during organogenesis. Gene Ontology (GO) annotation showed that the highest number of SE-related genes was involved in defense responses. The expression of fifteen selected SE-related candidate genes was assessed by RT-qPCR across nine centaury developmental stages, including embryogenic tissues. Notably, a newly reported transcript, named CeNA1, was specifically activated during embryogenic callus (ec) induction, making it a potential novel marker for early SE. These findings provide, for the first time, insight into SE-related transcriptional patterns, representing a step closer to uncovering the molecular basis of centaury’s developmental plasticity.
The Placenta as the Main Source of Serotonin in Ontogenetic Dynamics: Inflammation-Induced Modulation of Placental Serotonin Can Be Prevented by Immunoglobulin Administration
Authors: Bondarenko Nadezhda , Lifantseva Nadezhda , Voronova Svetlana , Melnikova Victoria
Published: December 2024

Placental serotonin is recognized as a key component of feto-placental physiology and can be influenced by environmental factors such as maternal diet, drugs, stress, and immune activation. In this study, we compared the contribution of placental and fetal sources to the maintenance of serotonin levels required for normal fetal development during ontogenetic dynamics. Our results demonstrated the leading role of the placenta at almost all stages of development. We investigated the modulatory effect of inflammation on placental serotonin levels. The data obtained showed that the susceptibility to prenatal inflammation depends on its severity and varies considerably at different stages of development. According to our results, inflammation-induced modulation of placental serotonin levels can be prevented by immunoglobulin administration at both early and late stages of development. Disturbances in placental serotonin signaling during critical developmental periods may have long-lasting consequences for the health and behavior of the offspring. Therefore, the ability to prevent environmental modulation of placental serotonin, and hence negative effects on the developing fetus, is of great importance.
Molecular Profiles of Sensitization to Non-Specific Lipid Transfer Proteins in Lithuania: Single Center Experience
Authors: Sakalauskaite Sandra , Pilkyte Ligita , Gasiuniene Edita , Gradauskiene Brigita
Published: December 2024

Non-specific Lipid Transfer proteins (nsLTPs) are relevant allergens of several pollens and plant foods. Sensitization to nsLTPs is not typical in our region. Still, it has become an increasingly common cause of IgE-mediated food allergies and food-induced anaphylaxis in Northern Europe in recent decades. No in-depth studies describe the prevalence of sensitization of molecular components to nsLTPs in Lithuania. This study aimed to determine the sensitization profile of atopic patients at the Immunology and Allergy Department of Kauno Klinikos to the components of nsLTPs, using molecular allergen component analysis. Sixty Lithuanian adults with symptoms of allergic rhinitis and/or allergic asthma and/or food allergies were included into the study. Specific immunoglobulin E (IgE) levels were measured using two in vitro techniques: allergen extract and molecular component analysis. Results showed that 25% of subjects were sensitized to nsLTP-containing allergen sources, mostly to Zea m 14, Mal d 3, Vit v 1, and Art v 3. The median amount of total IgE was higher in nsLTP-sensitized patients than in nsLTP-nonsensitized patients. Based on Cohen’s Kappa and McNemar tests, the results of allergen extract and component analysis tests do not always agree, especially when we determine the sensitization to allergen sources containing nsLTPs. Molecular allergen component analysis could be the first choice in determining detailed sensitization to nsLTPs in patients who experienced anaphylaxis of unknown origin.
Bombyx mori Metal Carboxypeptidases12 (BmMCP12) Is Involved in Host Protection Against Viral Infection
Authors: Tang Liang , Wei Qiong-Qiong , Xiao Yu , Tang Ming-Yan , Zhu Yan , Jiang Man-Gui , Chen Peng , Pan Zhi-Xin
Published: December 2024

Baculoviruses, the largest studied insect viruses, are highly pathogenic to host insects. Bombyx mori nucleopolyhedrovirus (BmNPV) is the main cause of nuclear polyhedrosis of silkworm, a viral disease that causes significant economic losses to the sericulture industry. The anti-BmNPV mechanism of the silkworm has not yet been characterized. Carboxypeptidase is an enzyme that is involved in virtually all life activities of animals and plants. Studies have shown that the carboxypeptidase family is related to insect immunity. There are few reports on the role of carboxypeptidase in the defense of silkworms against pathogen invasion. In this study, we identified the homologous gene Bombyx mori metal carboxypeptidases12 (BmMCP12) related to mammalian carboxypeptidase A2 (CPA2) and found that BmMCP12 had a Zn-pept domain. The BmMCP12 gene was primarily located in the cytoplasm and was highly expressed in the midgut of silkworms, and the expression level in BmN-SWU1 cells was upregulated after infection with BmNPV. After overexpression of the BmMCP12 gene, quantitative real-time (qRT)-PCR and Western blots showed that BmMCP12 could inhibit BmNPV replication, whereas knockout of the gene had the opposite effect. In addition, we constructed transgenic silkworm strains with a knockout of BmMCP12, and the transgenic strains had reduced resistance to BmNPV. These findings deepen the functional study of silkworm carboxypeptidase and provide a new target for BmNPV disease prevention in silkworms.
Angiopoietin-2: A Therapeutic Target for Vascular Protection in Hutchinson–Gilford Progeria Syndrome
Authors: Vakili Sahar , Cao Kan
Published: December 2024

Hutchinson–Gilford progeria syndrome (HGPS) is a pediatric condition characterized by clinical features that resemble accelerated aging. The abnormal accumulation of a toxic form of the lamin A protein known as progerin disrupts cellular functions, leading to various complications, including growth retardation, loss of subcutaneous fat, abnormal skin, alopecia, osteoporosis, and progressive joint contractures. Death primarily occurs as the result of complications from progressive atherosclerosis, especially from cardiac disease, such as myocardial infarction or heart failure, or cerebrovascular disease like stroke. Despite the availability of lonafarnib, the only US Food and Drug Administration-approved treatment for HGPS, cardiovascular complications remain the leading cause of morbidity and mortality in affected patients. Defective angiogenesis—the process of forming new blood vessels from existing ones—plays a crucial role in the development of cardiovascular disease. A recent study suggests that Angiopoietin-2 (Ang2), a pro-angiogenic growth factor that regulates angiogenesis and vascular stability, may offer therapeutic potential for the treatment of HGPS. In this review, we describe the clinical features and key cellular processes impacted by progerin and discuss the therapeutic potential of Ang2 in addressing these challenges.
Boosting Lipofection Efficiency Through Enhanced Membrane Fusion Mechanisms
Authors: Pavlov Rais V. , Akimov Sergey A. , Dashinimaev Erdem B. , Bashkirov Pavel V.
Published: December 2024

Gene transfection is a fundamental technique in the fields of biological research and therapeutic innovation. Due to their biocompatibility and membrane-mimetic properties, lipid vectors serve as essential tools in transfection. The successful delivery of genetic material into the cytoplasm is contingent upon the fusion of the vector and cellular membranes, which enables hydrophilic polynucleic acids to traverse the hydrophobic barriers of two intervening membranes. This review examines the critical role of membrane fusion in lipofection efficiency, with a particular focus on the molecular mechanisms that govern lipoplex–membrane interactions. This analysis will examine the key challenges inherent to the fusion process, from achieving initial membrane proximity to facilitating final content release through membrane remodeling. In contrast to viral vectors, which utilize specialized fusion proteins, lipid vectors necessitate a strategic formulation and environmental optimization to enhance their fusogenicity. This review discusses recent advances in vector design and fusion-promoting strategies, emphasizing their potential to improve gene delivery yield. It highlights the importance of understanding lipoplex–membrane fusion mechanisms for developing next-generation delivery systems and emphasizes the need for continued fundamental research to advance lipid-mediated transfection technology.
Protective Action of Cannabidiol on Tiamulin Toxicity in Humans—In Vitro Study
Authors: Pankowska Eryka , Kończak Oliwia , Żakowicz Paula , Wojciechowicz Tatiana , Gogulski Maciej , Radko Lidia
Published: December 2024

The growing awareness and need to protect public health, including food safety, require a thorough study of the mechanism of action of veterinary drugs in consumers to reduce their negative impact on humans. Inappropriate use of veterinary drugs in animal husbandry, such as tiamulin, leads to the appearance of residues in edible animal tissues. The use of natural substances of plant origin, extracted from hemp (Cannabis sativa L.), such as cannabidiol (CBD), is one of the solutions to minimize the negative effects of tiamulin. This study aimed to determine the effect of CBD on the cytotoxicity of tiamulin in humans. The cytotoxic activity of tiamulin and the effect of its mixtures with CBD were tested after 72 h exposure to three human cell lines: SH-SY5Y, HepG2 and HEK-293. Cytotoxic concentrations (IC50) of the tested drug and in combination with CBD were assessed using five biochemical endpoints: mitochondrial and lysosomal activity, proliferation, cell membrane integrity and effects on DNA synthesis. Oxidative stress, cell death and cellular morphology were also assessed. The nature of the interaction between the veterinary drug and CBD was assessed using the combination index. The long-term effect of tiamulin inhibited lysosomal (SH-SY5SY) and mitochondrial (HepG2) activity and DNA synthesis (HEK-293). IC50 values for tiamulin ranged from 2.1 to >200 µg/mL (SH-SY5SY), 13.9 to 39.5 µg/mL (HepG2) and 8.5 to 76.9 µg/mL (HEK-293). IC50 values for the drug/CBD mixtures were higher. Reduced levels of oxidative stress, apoptosis and changes in cell morphology were demonstrated after exposure to the mixtures. Interactions between the veterinary drug and CBD showed a concentration-dependent nature of tiamulin in cell culture, ranging from antagonistic (low concentrations) to synergistic effects at high drug concentrations. The increased risk to human health associated with the presence of the veterinary drug in food products and the protective nature of CBD use underline the importance of these studies in food toxicology and require further investigation.
Indocyanine Blue (ICB) as a Functional Alternative to Indocyanine Green (ICG) for Enhanced 700 nm NIR Imaging
Authors: Yamashita Atsushi , Jang Paul , Bao Kai , Kashiwagi Satoshi , Frangioni John V. , Choi Hak Soo
Published: December 2024

Despite significant advancements in bioimaging technology, only a limited number of fluorophores are currently approved for clinical applications. Indocyanine green (ICG) is the first FDA-approved near-infrared (NIR) fluorophore and has significantly advanced clinical interventions over the past three decades. However, its single-channel imaging at 800 nm emission is often insufficient for capturing comprehensive diagnostic information during surgery. In this study, we evaluate indocyanine blue (ICB), an ICG analog with a shorter polymethine bridge, as a promising candidate for multi-channel NIR imaging. ICB demonstrated peak absorption and emission approximately 100 nm shorter than ICG in aqueous solutions, placing it within the 700 nm range of the NIR window. Furthermore, ICB exhibited favorable solubility and optical properties in aqueous environments, supporting its potential for in vivo imaging applications. Notably, ICB shows rapid systemic clearance, likely due to its lower molecular weight, which facilitates clear visualization in angiography, cholangiography, and lymph node mapping with minimal background interference. Additionally, dual-channel imaging of tumors and lymph nodes was achieved using a tumor-targeting fluorophore in conjunction with ICB, illustrating the potential for enhanced intraoperative guidance. ICB emitting at 700 nm, therefore, can be useful in NIR imaging, broadening the possibilities for improved diagnostic accuracy and therapeutic outcomes in clinical settings.
Investigation of Host–Guest Interactions in 2-Ureido-4-ferrocenylpyrimidine Derivatives
Authors: Váradi Márk , Keszei Soma J. , Gömöry Ágnes , Kovács Margit , Kégl Tamás , Fodor Lajos , Skoda-Földes Rita
Published: December 2024

In the present study, synthesis, conformational behavior, host–guest complex formation, and electrochemical properties of novel 6-substituted-2-ureido-4-ferrocenylpyrimidines were explored. A comprehensive NMR spectroscopic investigation was carried out to confirm the structure and conformational equilibrium of the ureidopyrimidines through studying the temperature- and concentration dependence of NMR spectra. Low-temperature NMR measurements were used to clarify structural changes inflicted by a 2,6-diaminopyridine guest. Association constant (Kassoc) values of host–guest complexes were calculated based on low-temperature titrations. It was shown that the introduction of a pyridin-2-yl substituent in the pyrimidine ring in host 10 induced a considerable change not only in the conformational equilibrium of the host itself but also in that of the host–guest complex. Geometries and relative stabilities of the conformers of host 10 as well as its host–guest complexes were determined by quantum chemical calculations. Electrochemical behavior of ureidopyrimidine hosts and host–guest complexes was investigated by cyclic voltammetry (CV) and linear sweep voltammetry (LSV) measurements. Two ureidopyrimidine derivatives were immobilized on the surface of spectral graphite electrodes, and their electrochemical response on the addition of 2,6-diaminopyridine was compared. These results also supported the importance of the pyridin-2-yl substituent in the efficient sensing of the guest.
LncRNA-MSTRG.19083.1 Targets NTRK2 as a miR-429-y Sponge to Regulate Circadian Rhythm via the cAMP Pathway in Yak Testis and Cryptorchidism
Authors: Li Tianan , Yan Qiu , Nan Jinghong , Huang Xue , Wang Ruiqing , Zhang Yong , Zhao Xingxu , Wang Qi
Published: December 2024

Long noncoding RNAs (LncRNAs) play essential roles in numerous biological processes in mammals, such as reproductive physiology and endocrinology. Cryptorchidism is a common male reproductive disease. Circadian rhythms are actively expressed in the reproductive system. In this study, a total of 191 LncRNAs were obtained from yak testes and cryptorchids. Then, we identified NTRK2’s relationship to circadian rhythm and behavioral processes. Meanwhile, the ceRNA (LncRNA-MSTRG.19083.1/miR-429-y/NTRK2) network was constructed, and its influence on circadian rhythm was revealed. The results showed that NTRK2 and LncRNA-MSTRG.19083.1 were significantly upregulated, and miR-429-y was obviously decreased in cryptorchid tissue; NTRK2 protein was mainly distributed in the Leydig cells of the testis. In addition, the upregulation of the expression level of miR-429-y resulted in the significant downregulation of LncRNA and NTRK2 levels, while the mRNA and protein levels of CREB, CLOCK, and BMAL1 were significantly upregulated; the knockdown of miR-429-y resulted in the opposite changes. Our findings suggested that LncRNA-MSTRG.19083.1 competitively binds to miR-429-y to target NTRK2 to regulate circadian rhythm through the cAMP pathway. Taken together, the results of our study provide a comprehensive understanding of how the LncRNA-miRNA-mRNA networks operate when yak cryptorchidism occurs. Knowledge of circadian-rhythm-associated mRNAs and LncRNAs could be useful for better understanding the relationship between circadian rhythm and reproduction.
Functional Roles of the Charged Residues of the C- and M-Gates in the Yeast Mitochondrial NAD+ Transporter Ndt1p
Authors: Miniero Daniela Valeria , Palmieri Ferdinando , Quadrotta Virginia , Polticelli Fabio , Palmieri Luigi , Monné Magnus
Published: December 2024

Mitochondrial carriers transport organic acids, amino acids, nucleotides and cofactors across the mitochondrial inner membrane. These transporters consist of a three-fold symmetric bundle of six transmembrane α-helices that encircle a pore with a central substrate binding site, whose alternating access is controlled by a cytoplasmic and a matrix gate (C- and M-gates). The C- and M-gates close by forming two different salt-bridge networks involving the conserved motifs [YF][DE]XX[KR] on the even-numbered and PX[DE]XX[KR] on the odd-numbered transmembrane α-helices, respectively. We have investigated the effects on transport of mutating the C-gate charged residues of the yeast NAD+ transporter Ndt1p and performed molecular docking with NAD+ and other substrates into structural models of Ndt1p. Double-cysteine substitutions and swapping the positions of the C-gate charged-pair residues showed that all of them contribute to the high transport rate of wild-type Ndt1p, although no single salt bridge is essential for activity. The in silico docking results strongly suggest that both the C-gate motif mutations and our previously reported M-gate mutations affect gate closing, whereas those of the M-gate also affect substrate binding, which is further supported by molecular dynamics. In particular, NAD+ most likely interferes with the cation-π interaction between R303-W198, which has been proposed to exist in the Ndt1p M-gate in the place of one of the salt bridges. These findings contribute to understanding the roles of the charged C- and M-gate residues in the transport mechanism of Ndt1p.
Immortalization of Mesenchymal Stem Cells for Application in Regenerative Medicine and Their Potential Risks of Tumorigenesis
Authors: Yamaguchi Natsuki , Horio Eri , Sonoda Jukito , Yamagishi Miu , Miyakawa Satomi , Murakami Fumihiro , Hasegawa Hideaki , Katahira Yasuhiro , Mizoguchi Izuru , Fujii Yasuyuki , Chikazu Daichi , Yoshimoto Takayuki
Published: December 2024

Regenerative medicine utilizes stem cells to repair damaged tissues by replacing them with their differentiated cells and activating the body’s inherent regenerative abilities. Mesenchymal stem cells (MSCs) are adult stem cells that possess tissue repair and regenerative capabilities and immunomodulatory properties with a much lower risk of tumorigenicity, making them a focus of numerous clinical trials worldwide. MSCs primarily exert their therapeutic effects through paracrine effects via secreted factors, such as cytokines and exosomes. This has led to increasing interest in cell-free therapy, where only the conditioned medium (also called secretome) from MSC cultures is used for regenerative applications. However, MSCs face certain limitations, including cellular senescence, scarcity, donor heterogeneity, complexity, short survival post-implantation, and regulatory and ethics hurdles. To address these challenges, various types of immortalized MSCs (ImMSCs) capable of indefinite expansion have been developed. These cells offer significant promise and essential tools as a reliable source for both cell-based and cell-free therapies with the aim of translating them into practical medicine. However, the process of immortalization, often involving the transduction of immortalizing genes, poses potential risks of genetic instability and resultant malignant transformation. Cell-free therapy is particularly attractive, as it circumvents the risks of tumorigenicity and ethical concerns associated with live cell therapies. Rigorous safety tests, such as monitoring chromosomal abnormalities, are critical to ensure safety. Technologies like inducible or suicide genes may allow for the controlled proliferation of MSCs and induce apoptosis after their therapeutic task is completed. This review highlights recent advancements in the immortalization of MSCs and the associated risks of tumorigenesis.
Essential Role of Cortactin in Myogenic Differentiation: Regulating Actin Dynamics and Myocardin-Related Transcription Factor A-Serum Response Factor (MRTFA-SRF) Signaling
Authors: Ly Quoc Kiet , Nguyen Mai Thi , Ngo Thanh Huu Phan , Lee Wan
Published: December 2024

Cortactin (CTTN) is an actin-binding protein regulating actin polymerization and stabilization, which are vital processes for maintaining skeletal muscle homeostasis. Despite the established function of CTTN in actin cytoskeletal dynamics, its role in the myogenic differentiation of progenitor cells remains largely unexplored. In this study, we investigated the role of CTTN in the myogenic differentiation of C2C12 myoblasts by analyzing its effects on actin cytoskeletal remodeling, myocardin-related transcription factor A (MRTFA) nuclear translocation, serum response factor (SRF) activation, expression of myogenic transcription factors, and myotube formation. CTTN expression declined during myogenic differentiation, paralleling the reduction in MyoD, suggesting a potential role in the early stages of myogenesis. We also found that CTTN knockdown in C2C12 myoblasts reduced filamentous actin, enhanced globular actin levels, and inhibited the nuclear translocation of MRTFA, resulting in suppressed SRF activity. This led to the subsequent downregulation of myogenic regulatory factors, such as MyoD and MyoG. Furthermore, CTTN knockdown reduced the nuclear localization of YAP1, a mechanosensitive transcription factor, further supporting its regulatory roles in cell cycle and proliferation. Consequently, CTTN depletion impeded proliferation, differentiation, and myotube formation in C2C12 myoblasts, highlighting its dual role in the coordination of cell cycle regulation and myogenic differentiation of progenitor cells during myogenesis. This study identifies CTTN as an essential regulator of myogenic differentiation via affecting the actin remodeling-MRTFA-SRF signaling axis and cell proliferation.