bluuebunny/crossref_metadata_2025 huggingface dataset:
On Observer and Controller Design for Nonlinear Hadamard Fractional-Order One-Sided Lipschitz Systems
Authors: Jmal Assaad , Naifar Omar , Rhaima Mohamed , Ben Makhlouf Abdellatif , Mchiri Lassaad
Published: October 2024

This paper presents an extensive investigation into the state feedback stabilization, observer design, and observer-based controller design for a specific category of nonlinear Hadamard fractional-order systems. The research extends the conventional integer-order derivative to the Hadamard fractional-order derivative, offering a more universally applicable method for system analysis. Furthermore, the traditional Lipschitz condition is adapted to a one-sided Lipschitz condition, broadening the range of systems amenable to analysis using these techniques. The efficacy of the proposed theoretical findings is illustrated through several numerical examples. For instance, in Example 1, we select an order of derivative r = 0.8; in Example 2, r is set to 0.9; and in Example 3, r = 0.95. This study enhances the comprehension and regulation of nonlinear Hadamard fractional-order systems, setting the stage for future explorations in this domain.
Simulation-Based Optimization Workflow of CO2-EOR for Hydraulic Fractured Wells in Wolfcamp A Formation
Authors: Bui Dung , Pham Duc , Nguyen Son , Nguyen Kien
Published: October 2024

Hydraulic fracturing has enabled production from unconventional reservoirs in the U.S., but production rates often decline sharply, limiting recovery factors to under 10%. This study proposes an optimization workflow for the CO2 huff-n-puff process for multistage-fractured horizontal wells in the Wolfcamp A formation in the Delaware Basin. The potential for enhanced oil recovery and CO2 sequestration simultaneously was addressed using a coupled geomechanics–reservoir simulation. Geomechanical properties were derived from a 1D mechanical earth model and integrated into reservoir simulation to replicate hydraulic fracture geometries. The fracture model was validated using a robust production history matching. A fluid phase behavior analysis refined the equation of state, and 1D slim tube simulations determined a minimum miscibility pressure of 4300 psi for CO2 injection. After the primary production phase, various CO2 injection rates were tested from 1 to 25 MMSCFD/well, resulting in incremental oil recovery ranging from 6.3% to 69.3%. Different injection, soaking and production cycles were analyzed to determine the ideal operating condition. The optimal scenario improved cumulative oil recovery by 68.8% while keeping the highest CO2 storage efficiency. The simulation approach proposed by this study provides a comprehensive and systematic workflow for evaluating and optimizing CO2 huff-n-puff in hydraulically fractured wells, enhancing the recovery factor of unconventional reservoirs.
Chromosomal Localization and Diversity Analysis of 5S and 18S Ribosomal DNA in 13 Species from the Genus Ipomoea
Authors: Wu Jingyu , Lang Tao , Zhang Cong , Yang Fan , Yang Feiyang , Qu Huijuan , Pu Zhigang , Feng Junyan
Published: October 2024

Background: Sweet potato (Ipomoea batatas (L.) Lam.), a key global root crop, faces challenges due to its narrow genetic background. This issue can be addressed by utilizing the diverse genetic resources of sweet potato’s wild relatives, which are invaluable for its genetic improvement. Methods: The morphological differences in leaves, stems, and roots among 13 Ipomoea species were observed and compared. Chromosome numbers were determined by examining metaphase cells from root tips. Fluorescence in situ hybridization (FISH) was used to identify the number of 5S and 18S rDNA sites in these species. PCR amplification was performed for both 5S and 18S rDNA, and phylogenetic relationships among the species were analyzed based on the sequences of 18S rDNA. Results: Three species were found to have enlarged roots among the 13 Ipomoea species. Chromosome analysis revealed that I. batatas had 90 chromosomes, Ipomoea pes-tigridis had 28 chromosomes, while the remaining species possessed 30 chromosomes. Detection of rDNA sites in the 13 species showed two distinct 5S rDNA site patterns and six 18S rDNA site patterns in the 12 diploid species. These rDNA sites occurred in pairs, except for the seven 18S rDNA sites observed in Ipomoea digitata. PCR amplification of 5S rDNA identified four distinct patterns, while 18S rDNA showed only a single pattern across the species. Phylogenetic analysis divided the 13 species into two primary clades, with the closest relationships found between I. batatas and Ipomoea trifida, as well as between Ipomoea platensis and I. digitata. Conclusions: These results enhance our understanding of the diversity among Ipomoea species and provide valuable insights for breeders using these species to generate improved varieties.
GWAS for Drought Resilience Traits in Red Clover (Trifolium pratense L.)
Authors: Vleugels Tim , Ruttink Tom , Ariza-Suarez Daniel , Dubey Reena , Saleem Aamir , Roldán-Ruiz Isabel , Muylle Hilde
Published: October 2024

Red clover (Trifolium pratense L.) is a well-appreciated grassland crop in temperate climates but suffers from increasingly frequent and severe drought periods. Molecular markers for drought resilience (DR) would benefit breeding initiatives for red clover, as would a better understanding of the genes involved in DR. Two previous studies, as follows, have: (1) identified phenotypic DR traits in a diverse set of red clover accessions; and (2) produced genotypic data using a pooled genotyping-by-sequencing (GBS) approach in the same collection. In the present study, we performed genome-wide association studies (GWAS) for DR using the available phenotypic and genotypic data. Single nucleotide polymorphism (SNP) calling was performed using GBS data and the following two red clover genome assemblies: the recent HEN-17 assembly and the Milvus assembly. SNP positions with significant associations were used to delineate flanking regions in both genome assemblies, while functional annotations were retrieved from Medicago truncatula orthologs. GWAS revealed 19 significant SNPs in the HEN-17-derived SNP set, explaining between 5.3 and 23.2% of the phenotypic variation per SNP–trait combination for DR traits. Among the genes in the SNP-flanking regions, we identified candidate genes related to cell wall structuring, genes encoding sugar-modifying proteins, an ureide permease gene, and other genes linked to stress metabolism pathways. GWAS revealed 29 SNPs in the Milvus-derived SNP set that explained substantially more phenotypic variation for DR traits, between 5.3 and 42.3% per SNP–trait combination. Candidate genes included a DEAD-box ATP-dependent RNA helicase gene, a P-loop nucleoside triphosphate hydrolase gene, a Myb/SANT-like DNA-binding domain protein, and an ubiquitin–protein ligase gene. Most accessions in this study are genetically more closely related to the Milvus genotype than to HEN-17, possibly explaining how the Milvus-derived SNP set yielded more robust associations. The Milvus-derived SNP set pinpointed 10 genomic regions that explained more than 25% of the phenotypic variation for DR traits. A possible next step could be the implementation of these SNP markers in practical breeding programs, which would help to improve DR in red clover. Candidate genes could be further characterized in future research to unravel drought stress resilience in red clover in more detail.
Transcriptome Analysis Deciphers the Underlying Molecular Mechanism of Peanut Lateral Branch Angle Formation Using Erect Branching Mutant
Authors: He Liangqiong , Yu Conghui , Wang Guanghao , Su Lei , Xing Xin , Liu Tiantian , Huang Zhipeng , Xia Han , Zhao Shuzhen , Gao Zhongkui , Wang Xingjun , Zhao Chuanzhi , Han Zhuqiang , Pan Jiaowen
Published: October 2024

Background The growth habit (GH), also named the branching habit, is an important agronomic trait of peanut and mainly determined by the lateral branch angle (LBA). The branching habit is closely related to peanut mechanized farming, pegging, yield, and disease management. Objectives However, the molecular basis underlying peanut LBA needs to be uncovered. Methods In the present study, an erect branching peanut mutant, eg06g, was obtained via 60Co γ-ray-radiating mutagenesis of a spreading-type peanut cultivar, Georgia-06G (G06G). RNA-seq was performed to compare the transcriptome variation of the upper sides and lower sides of the lateral branch of eg06g and G06G. Results In total, 4908 differentially expressed genes (DEGs) and 5833 DEGs were identified between eg06g and G06G from the lower sides and upper sides of the lateral branch, respectively. GO, KEGG, and clustering enrichment analysis indicated that the carbohydrate metabolic process, cell wall organization or biogenesis, and plant hormone signal transduction were mainly enriched in eg06g. Conclusions Further analysis showed that the genes involved in starch biosynthesis were upregulated in eg06g, which contributed to amyloplast sedimentation and gravity perception. Auxin homeostasis and transport-related genes were found to be upregulated in eg06g, which altered the redistribution of auxin in eg06g and in turn triggered apoplastic acidification and activated cell wall modification-related enzymes, leading to tiller angle establishment through the promotion of cell elongation at the lower side of the lateral branch. In addition, cytokinin and GA also demonstrated synergistic action to finely regulate the formation of peanut lateral branch angles. Collectively, our findings provide new insights into the molecular regulation of peanut LBA and present genetic materials for breeding peanut cultivars with ideotypes.
Non-Invasive Prenatal Screening from a Genetic Counseling Prospective: Pre and Post-Genetic Counseling Regarding Rare Chromosomal Abnormalities and Incidental Finding
Authors: Matteo Della Monica , Lorenzo Cipriano , Raffaele Piscopo , Carmelo Piscopo
Published: October 2024

Background: Arising in the late 1990s, when a promising role in prenatal diagnostics was first delineated for circulating fetal DNA, non-invasive prenatal tests (NIPTs) have been increasingly used with more frequency and popularity. These exams have been used as a prenatal screening tests for genetic diseases. Initially, they were developed for the investigation of the main fetal chromosomal aneuploidies, but lately they have also been used to rule out genomic microrearrangements and monogenic conditions. However, along with great opportunities and potential, the tests can show inconclusive or unexpected results. Several studies have shown that the current pre-test counseling is often insufficient, and more oriented at providing pieces of information about the identifiable diseases rather than providing extensive information on all possible scenarios which may affect both the fetus and the pregnant mother, especially in the case of an invasive test for the pregnant mother. Methods and Results: We have gathered from the literature on NIPT the main pitfalls, imperfections, and particular cases associated with this innovative diagnostic procedure. Conclusions: In view of further improvements in the methods that can limit the inconclusive or unexpected results, this paper aims to reinforce the importance of more accurate pre-test counseling with comprehensive information about the above-mentioned questions, as well as ultrasound use and also the creation of an international consensus statement concerning these topics.
Capitalizing on Animality: Monstrosity and Multispecies Relations in Jordan Peele’s Nope (2022)
Authors: King Heather
Published: October 2024

One amongst many of the defining characteristics of so-called ‘late stage’ capitalism are human-animal relationships that have become acrimonious, hostile, or even monstrous in nature. A foundational premise of monster theory, and one that Jeffrey Jerome Cohen’s seminal 1996 edited collection of the same name suggests, is that the construction of the monster in popular culture is fraught with the boundaries that constitute the society that has spawned them; the monstrous body “exists only to be read” (p. 4). Bringing together the theoretical insights of the Marxist theory of reification, critical animal studies, and monster theory, this article examines the ways in which cinematic depictions of gigantic monstrosity can inform our theorizing of multispecies relationships under capitalism. Specifically, I explore how the tensions between capital and human-animal relationships serve to construct and constitute the multiform monster, Jean Jacket, in Jordan Peele’s 2022 film Nope. Through an examination of the multispecies relationalities that the film portrays, I argue that the figure of Jean Jacket is a monstrous culmination of the reified and therefore, necessarily deferred nature of human-animal relationships under capital. However, Nope’s conclusion alerts us to the radical dereifying potential of multispecies bonds of care and embodied knowledge; systems of resistance that can be forged even within our current capitalist ruins.
Volatile Constituents of Cymbopogon citratus (DC.) Stapf Grown in Greenhouse in Serbia: Chemical Analysis and Chemometrics
Authors: Aćimović Milica , Lončar Biljana , Todosijević Marina , Lekić Stefan , Erceg Tamara , Pezo Milada , Pezo Lato
Published: October 2024

The present study investigated the volatile constituents of Cymbopogon citratus (lemongrass) grown in a greenhouse environment in Serbia, marking the first commercial cultivation of the plant for essential oil production in the region. The essential oils and hydrolates obtained through steam distillation were analyzed via gas chromatography–mass spectrometry (GC-MS), and the resulting chemical data were further processed using chemometric methods. This study applied quantitative structure retention relationship (QSRR) analysis, employing molecular descriptors (MDs) and artificial neural networks (ANNs) to predict the retention indices (RIs) of the compounds. A genetic algorithm (GA) was used to select the most relevant MDs for this predictive modeling. A total of 29 compounds were annotated in the essential oils, with geranial and neral being the dominant components, while 37 compounds were detected in the hydrolates. The ANN models effectively predicted the RIs of both essential oils and hydrolates, demonstrating high statistical accuracy and low prediction errors. This research offers valuable insights into the chemical profile of lemongrass cultivated in temperate conditions and advances QSRR modeling for essential oil analysis.
Estimation of Cucumber Fruit Yield Cultivated Under Different Light Conditions in Greenhouses
Authors: Hong Inseo , Yu Jin , Hwang Seung Jae , Kwack Yurina
Published: October 2024

In recent years, an increase in the frequency of low-sunlight conditions due to climate change has resulted in a decline in the yield and quality of crops for greenhouse farmers, leading to significant challenges in maintaining optimal plant growth. The crop growth model can be used to predict changes in cucumber yield in response to variations in sunlight, which can help efficiently address sunlight shortages. The objective of this study was to improve and validate the model for predicting cucumber yield under different light environment conditions, including shading and supplemental lighting. The model comprises three steps: LAI prediction, daily assimilate yield prediction, and fruit yield prediction, each of which involves modifying the coefficients applied to suit the cucumber cultivar and environment condition. The improved model demonstrated a high degree of accuracy in predicting cucumber yields in the control and low-sunlight treatments (10, 20, and 30% shading), with a coefficient of determination (R2) > 0.98. When supplemental lighting was incorporated into the control and shading treatments, the accuracy of the improved model in predicting cucumber yield was also high, with a coefficient of determination (R2) > 0.99. The model also accurately predicted the decrease in cucumber fruit yield under low-sunlight conditions (shading treatments) and the increase in yield due to supplemental lighting. The findings of this study indicate that the improved cucumber yield prediction model can be applied to assess the efficacy of yield reduction in low-sunlight conditions and the potential for yield enhancement through supplemental lighting.
Application of Silicon Iron and Silver Nanoparticles Improve Vegetative Development and Physiological Characteristics of Boysenberry Plants Grown under Salinity Stress In Vitro Cultivation Conditions
Authors: Kurt Zehra , Ateş Sevinç
Published: October 2024

Salinity is one of the most important abiotic stress factors that affect plant growth and limit agricultural productivity. In this study, the effects of iron (FeNP), silver (AgNP), and silicon dioxide (SiNP) nanoparticles on the morphological and physiological parameters of in vitro boysenberry plants grown under salinity stress (NaCl) were investigated. According to our study results, higher values were obtained from SiNP application in terms of shoot development parameters; FeNP application was found to be more successful for root development; AgNP application was effective in terms of SPAD, leaf relative water content (LRWC), and relative growth rate (RGR); and FeNP application increased superoxide dismutase (SOD) and catalase (CAT) enzyme activities. Salt stress significantly affected root development, SPAD values, LRWC and RGR, and SOD and CAT enzyme activities. As a result, under salt stress conditions, SiNP, FeNP, and AgNP applications can significantly reduce the negative effects of stress and promote the vegetative development of the plant compared to control conditions.
Early Generation Selection of Potato Breeding Lines
Authors: Sapakhova Zagipa , Abilda Zhanar , Toishimanov Maxat , Daurov Dias , Daurova Ainash , Raissova Nurgul , Sidorik Alexander , Kanat Rakhim , Zhambakin Kabyl , Shamekova Malika
Published: October 2024

Potato is the third most important food crop in the world in terms of adaptability, yield potential, and nutritional advantages. This study aimed to conduct potato breeding work for cultivation in Kazakhstan; potato breeding for further processing into chips with cultivation in the northern regions; and the selection of potatoes for processing into frozen French fries with cultivation in the southern and south-eastern regions. Potato varieties (Fontane, Santana, and Punchy) were used as reference varieties to check molecular markers linked to maturity, tuber shape, and flesh color. A total of 42 potato breeding lines crossed from Yagodnyi 19 and CIP clone 397079-6 were used in this study to identify prospective breeding lines. The research was carried out between 2023 and 2024 and under greenhouse conditions. According to the results of molecular analysis and phenotypic data, 21 breeding lines were identified as prospective potato breeding lines. The majority of these potato breeding lines had a round tuber shape and were recommended for chip processing. Three breeding lines had a long oval tuber shape, making them suitable for French fries. Six breeding lines with short-oval and oval tuber shapes were found for consumer potato processing. It is recommended that the breeding process and studies of biochemical properties are continued in all of these identified potato breeding lines.
The Effects of Green Spaces and Noise Exposure on the Risk of Ischemic Stroke: A Case–Control Study in Lebanon
Authors: El Masri Jad , Finge Hani , Afyouni Ahmad , Baroud Tarek , Ajaj Najla , Ghazi Maya , El Masri Diala , Younes Mahmoud , Salameh Pascale , Hosseini Hassan
Published: October 2024

Background: Environmental surroundings reduce the rate of several diseases, especially those related to stressful events. Ischemic stroke can be affected by such events, either directly or through its risk factors. Therefore, the present study evaluates the effects of green spaces and noise exposure on the risk of ischemic stroke. Methods: A case–control study was carried out, including 200 ischemic stroke cases within the first 48 h of diagnosis and 200 controls, divided equally into hospitalized and non-hospitalized participants. Controls were matched to cases based on age and gender. Socio-demographic characteristics were assessed, in addition to environmental surroundings and noise exposure at home and at workplaces. Results: Living in a house, having a house garden, and taking care of the garden were associated with a lower risk of suffering an ischemic stroke (p < 0.001, p < 0.001, and p = 0.009, respectively). However, having buildings as the view from home led to a higher stroke rate (p < 0.001). Working in an urban area, the workplace being surrounded by buildings, and the workplace not being surrounded by green spaces were also associated with a higher risk of suffering an ischemic stroke (p = 0.002, p = 0.001, and p = 0.03, respectively). As for noise exposure, being exposed to traffic noise, human noise, and other types of noise was significantly associated with a higher risk of ischemic stroke, while being exposed to higher levels of natural noise was significantly associated with a lower risk of ischemic stroke. Higher levels of noise were also associated with higher risks of ischemic stroke in homes and workplaces (p < 0.001 and p = 0.008, respectively). Conclusions: Environmental surroundings and noise exposure were found to affect the risk of ischemic stroke. Greater green spaces and lower noise exposure play a protective role against ischemic stroke, suggesting a possible prevention strategy through environmental modifications at home and workplaces.
Online and Face-to-Face Mat Pilates Training for Long COVID-19 Patients: A Randomized Controlled Trial on Health Outcomes
Authors: Cunha Ana Clara Ribeiro , Silva Juliana Cristina , Garcês Caroline Pereira , Sisconeto Tássia Magnabosco , Nascimento João Luiz Rezende , Amaral Ana Luiza , Cunha Thulio Marquez , Mariano Igor Moraes , Puga Guilherme Morais
Published: October 2024

This study investigated the impacts of online and face-to-face Mat Pilates training in adults with persistent symptoms of long COVID on health outcomes. Forty-nine patients (52 ± 5.85 yr.) diagnosed with long COVID related to fatigue symptoms were randomly included in three groups: online Mat Pilates training (n = 16), face-to-face Mat Pilates training (n = 15), and a control group (n = 18) without training. Mat Pilates training was conducted three times a week for 12 weeks. Fatigue, functional capacity, anthropometrics, body composition, and cardiometabolic markers were assessed before and after the interventions. Two-factor Generalized Estimating Equation analyses identified significant differences with Bonferroni post hoc testing (p < 0.05). After the intervention, only the face-to-face Mat Pilates training group had an improved total, physical and mental fatigue, trunk isometric strength, upper limb muscle endurance strength, and aerobic capacity (p < 0.05). No changes were found in fat mass, muscle mass, free fat mass, % of fat, body mass, body mass index, or waist and hip circumferences. No significant changes were observed in blood glucose, glycated hemoglobin, triglycerides, total cholesterol, high-density lipoprotein, low-density lipoprotein, or blood pressure (p > 0.05). Our results highlight the potential of face-to-face Mat Pilates training as an effective intervention to mitigate persistent symptoms of long COVID related to fatigue and functional capacities.
Polypharmacy in Older Patients with Multimorbidity: The Agreement Between Patient and General Practitioner-Reported Drugs Observed in a Pilot cRCT
Authors: Schäfer Lena , Paulitsch Michael , Hanf Maria , Dinh Truc Sophia , Klein Astrid-Alexandra , Klasing Sophia , Seidling Hanna , Voigt Karen , van den Akker Marjan
Published: October 2024

Polypharmacy (≥5 drugs) increases the risk of discrepancies between patient- and general practitioner (GP)-reported drugs, leading to adverse outcomes. This explorative analysis assesses the agreement between patient- and GP-reported drugs under the influence of a paper-based patient portfolio in a pilot cluster randomized controlled trial (cRCT). Complete data were available for 68 patients aged 65 or older (26 were female), with multimorbidity, polypharmacy, and at least one hospitalization in the past year. Agreement was assessed for drug name and strength level. Differences between the intervention and control group (IG/CG) and comparisons between two time points (six-month interval) stratified according to gender were analyzed using Wilcoxon and Mann–Whitney U tests (α = 5%). To evaluate the reasons for discrepancies, the agreement of active pharmaceutical ingredients (APIs) and anatomical therapeutic chemical (ATC) groups was analyzed. At baseline, the agreement was 72.1% for the IG and 73.9% for the CG. Inclusion of the reported drug strength reduced the agreement in both groups (IG 66.7%, CG 60.0%). Agreement for the IG decreased statistically significantly after six months (−5.4%). ATC groups B, C, and H had the highest agreement, while N, R, and Z had the lowest. Large discrepancies in the drugs reported, due to the APIs and the corresponding ATC group, were observed.
Role of Exogenous Pyruvate in Maintaining Adenosine Triphosphate Production under High-Glucose Conditions through PARP-Dependent Glycolysis and PARP-Independent Tricarboxylic Acid Cycle
Authors: Yako Hideji , Niimi Naoko , Takaku Shizuka , Kato Ayako , Kato Koichi , Sango Kazunori
Published: October 2024

Pyruvate serves as a key metabolite in energy production and as an anti-oxidant. In our previous study, exogenous pyruvate starvation under high-glucose conditions induced IMS32 Schwann cell death because of the reduced glycolysis–tricarboxylic acid (TCA) cycle flux and adenosine triphosphate (ATP) production. Thus, this study focused on poly-(ADP-ribose) polymerase (PARP) to investigate the detailed molecular mechanism of cell death. Rucaparib, a PARP inhibitor, protected Schwann cells against cell death and decreased glycolysis but not against an impaired TCA cycle under high-glucose conditions in the absence of pyruvate. Under such conditions, reduced pyruvate dehydrogenase (PDH) activity and glycolytic and mitochondrial ATP production were observed but not oxidative phosphorylation or the electric transfer chain. In addition, rucaparib supplementation restored glycolytic ATP production but not PDH activity and mitochondrial ATP production. No differences in the increased activity of caspase 3/7 and the localization of apoptosis-inducing factor were found among the experimental conditions. These results indicate that Schwann cells undergo necrosis rather than apoptosis or parthanatos under the aforementioned conditions. Exogenous pyruvate plays a pivotal role in maintaining the flux in PARP-dependent glycolysis and the PARP-independent TCA cycle in Schwann cells under high-glucose conditions.
Human iPSCs from Aged Donors Retain Their Mitochondrial Aging Signature
Authors: Lejri Imane , Cader Zameel , Grimm Amandine , Eckert Anne
Published: October 2024

Aging represents the leading risk factor for developing neurodegenerative disorders. One of the nine hallmarks of aging is mitochondrial dysfunction. Age-related mitochondrial alterations have been shown to affect mitochondrial energy metabolism, reduction-oxidation homeostasis, and mitochondrial dynamics. Previous reports have shown that induced pluripotent stem cells (iPSCs) from aged donors do not keep the aging signature at the transcriptomic level. However, not all aspects of aging have been investigated, and especially not the mitochondria-related aging signature. Therefore, the present study compared the mitochondrial function in iPSCs from healthy aged donors compared to those of young donors. We addressed whether aged iPSCs may be used as drug-screening models of “aging in a dish” to identify therapies alleviating mitochondria aging. Compared to iPSCs from young donors, we demonstrate that iPSCs from aged donors show impaired mitochondrial bioenergetics and exhibit a rise in reactive oxygen species generation. Furthermore, aged iPSCs present a lower mitochondrial mass and alterations in the morphology of the mitochondrial network when compared to iPSCs from young donors. This study provides the first evidence that the aging phenotype is present at the mitochondrial level in iPSCs from aged donors, ranging from bioenergetics to mitochondrial network morphology. This model might be used to screen mitochondria-targeting drugs to promote healthy aging at the mitochondrial level.
Effect of Benzyl Alcohol on Main Defense System Components of Galleria mellonella (Lepidoptera)
Authors: Kazek Michalina , Kaczmarek Agata , Wrońska Anna K. , Boguś Mieczysława I.
Published: October 2024

Benzyl alcohol (E1519) is an aromatic alcohol used in the pharmaceutical and food industry. It is used to protect food products against microorganisms during storage, as a flavoring in the production of chocolate and confectionery products, as an important ingredient in fragrance, and as a preservative in medical products. However, little is known of its effect on insects. The main aim of this study was to determine the influence of benzyl alcohol on the defense systems of the wax moth Galleria mellonella, i.e., its cuticular lipid composition and critical elements of its immune system. A gas chromatography/mass spectrometry (GC/MS) analysis found benzyl alcohol treatment to elicit significant quantitative and qualitative differences in cuticular free fatty acid (FFA) profiles. Our findings indicate that benzyl alcohol treatment increased the levels of HSP70 and HSP90 and decreased those of HSF1, histamine, and cysteinyl leukotriene. Benzyl alcohol application also increased dismutase level in the hemolymph and lowered those of catalase and 8-OHdG. The treatment also had negative effects on G. mellonella hemocytes and a Sf9 cell line in vitro: 48-h treatment resulted in morphological changes, with the remaining cells being clearly spindle-shaped with numerous granules. The high insecticidal activity of compound and its lack of toxicity towards vertebrates suggest it could be an effective insecticide.
High-Fat Diet-Induced Blood–Brain Barrier Dysfunction: Impact on Allodynia and Motor Coordination in Rats
Authors: Ubaldo-Reyes Laura M. , Espitia-Bautista Estefania , Barajas-Martínez Antonio , Martínez-Tapia Ricardo , Rodríguez-Mata Verónica , Noriega-Navarro Roxana , Escalona Rene , Castillo-Hernández Jesús , Pérez-Torres Armando , Navarro Luz
Published: October 2024

The associations among increased pain sensitivity, obesity, and systemic inflammation have not been described as related to BBB dysfunctions. To analyze the metabolic, behavioral, and inflammatory effects of a high-fat diet (HFD) and ultrastructural modifications in brain regions, we used an in vivo experimental model. Adult male Wistar rats were randomly assigned to one of two conditions, an ad libitum control group or an HFD (60%)-fed group, for eight weeks. At the end of the protocol, glucose and insulin tolerance tests were performed. Additionally, we analyzed the response to a normally innocuous mechanical stimulus and changes in motor coordination. At the end of the protocol, HFD-fed rats presented increased HOMA–IR and metabolic syndrome (MetS) prevalence. HFD-fed rats also developed an increased nociceptive response to mechanical stimuli and neurological injury, resulting in impaired motor function. Hypothalamus and cerebellum neurons from HFD-fed rats presented with nuclear swelling, an absence of nucleoli, and karyolysis. These results reveal that HFD consumption affects vital brain structures such as the cerebellum, hippocampus, and hypothalamus. This, in turn, could be producing neuronal damage, impairing cellular communication, and consequently altering motricity and pain sensitivity. Although direct evidence of a causal link between BBB dysfunction and sensory-motor changes was not observed, understanding the association uncovered in this study could lead to targeted therapeutic strategies.
Analysis of Azathioprine Metabolites in Autoimmune Hepatitis Patient Blood—Method Development and Validation
Authors: Guba Andrea , Kováts Patrícia , Mezei Zoltán A. , Papp Mária , Csősz Éva , Kalló Gergő
Published: October 2024

Autoimmune hepatitis (AIH) is a chronic inflammatory liver disease treated by steroids and immunomodulator thiopurine drugs such as azathioprine (AZA). AZA is metabolized in the human body into bioactive forms such as 6-thioguanine (6-TG) and 6-methyl-mercaptopurine (6-MMP). Monitoring the levels of bioactive AZA metabolites is very important for proper treatment of patients. In this study, our aim was to develop and validate a fast and sensitive ultra-high performance liquid chromatography–mass spectrometry (UHPLC–MS) method for the analysis of 6-TG and 6-MMP from blood samples of patients with AIH to monitor the level of these bioactive metabolites. The detection and quantification of the analytes was carried out by Selected Reaction Monitoring (SRM)-based targeted mass spectrometry. The method was validated according to the EMA guidelines. Blood samples from patients with AIH treated with AZA were analysed with the developed method. The method was successfully validated with appropriate accuracy and precision for the target biomolecules and their concentration in the samples from patients with AIH was determined. The developed and validated UHPLC–MS method enables the fast and precise analysis of AZA metabolites.
The Large GTPase Guanylate-Binding Protein-1 (GBP-1) Promotes Mitochondrial Fission in Glioblastoma
Authors: Kalb Ryan C. , Nyabuto Geoffrey O. , Morran Michael P. , Maity Swagata , Justinger Jacob S. , Nestor-Kalinoski Andrea L. , Vestal Deborah J.
Published: October 2024

Glioblastomas (aka Glioblastoma multiformes (GBMs)) are the most deadly of the adult brain tumors. Even with aggressive treatment, the prognosis is extremely poor. The large GTPase Guanylate-Binding Protein-1 (GBP-1) contributes to the poor prognosis of GBM by promoting migration and invasion. GBP-1 is substantially localized to the cytosolic side of the outer membrane of mitochondria in GBM cells. Because mitochondrial dynamics, particularly mitochondrial fission, can drive cell migration and invasion, the potential interactions between GBP-1 and mitochondrial dynamin-related protein 1 (Drp1) were explored. Drp1 is the major driver of mitochondrial fission. While GBP-1 and Drp1 both had punctate distributions within the cytoplasm and localized to regions of the cytoplasmic side of the plasma membrane of GBM cells, the proteins were only molecularly co-localized at the mitochondria. Subcellular fractionation showed that the presence of elevated GBP-1 promoted the movement of Drp1 from the cytosol to the mitochondria. The migration of U251 cells treated with the Drp1 inhibitor, Mdivi-1, was less inhibited in the cells with elevated GBP-1. Elevated GBP-1 in GBM cells resulted in shorter and wider mitochondria, most likely from mitochondrial fission. Mitochondrial fission can drive several important cellular processes, including cell migration, invasion, and metastasis.
Downregulation of the GhROD1 Gene Improves Cotton Fiber Fineness by Decreasing Acyl Pool Saturation, Stimulating Small Heat Shock Proteins (sHSPs), and Reducing H2O2 Production
Authors: Ding Bo , Liu Bi , Zhu Xi , Zhang Huiming , Hu Rongyu , Li Silu , Zhang Liuqin , Jiang Linzhu , Yang Yang , Zhang Mi , Zhao Juan , Pei Yan , Hou Lei
Published: October 2024

Cotton fiber is one of the most important natural fiber sources in the world, and lipid metabolism plays a critical role in its development. However, the specific role of lipid molecules in fiber development and the impact of fatty acid alterations on fiber quality remain largely unknown. In this study, we demonstrate that the downregulation of GhROD1, a gene encoding phosphatidylcholine diacylglycerol cholinephosphotransferase (PDCT), results in an improvement of fiber fineness. We found that GhROD1 downregulation significantly increases the proportion of linoleic acid (18:2) in cotton fibers, which subsequently upregulates genes encoding small heat shock proteins (sHSPs). This, in turn, reduces H2O2 production, thus delaying secondary wall deposition and leading to finer fibers. Our findings reveal how alterations in linoleic acid influence cellulose synthesis and suggest a potential strategy to improve cotton fiber quality by regulating lipid metabolism pathways.
Monitoring Myelin Lipid Composition and the Structure of Myelinated Fibers Reveals a Maturation Delay in CMT1A
Authors: Capodivento Giovanna , Camera Mattia , Liessi Nara , Trada Anna , Debellis Doriana , Schenone Angelo , Armirotti Andrea , Visigalli Davide , Nobbio Lucilla
Published: October 2024

Findings accumulated over time show that neurophysiological, neuropathological, and molecular alterations are present in CMT1A and support the dysmyelinating rather than demyelinating nature of this neuropathy. Moreover, uniform slowing of nerve conduction velocity is already manifest in CMT1A children and does not improve throughout their life. This evidence and our previous studies displaying aberrant myelin composition and structure in adult CMT1A rats prompt us to hypothesize a myelin and axon developmental defect in the CMT1A peripheral nervous system. Peripheral myelination begins during the early stages of development in mammals and, during this process, chemical and structural features of myelinated fibers (MFs) evolve towards a mature phenotype; deficiencies within this self-modulating circuit can cause its blockage. Therefore, to shed light on pathophysiological mechanisms that occur during development, and to investigate the relationship among axonal, myelin, and lipidome deficiencies in CMT1A, we extensively analyzed the evolution of both myelin lipid profile and MF structure in WT and CMT1A rats. Lipidomic analysis revealed a delayed maturation of CMT1A myelin already detectable at P10 characterized by a deprivation of sphingolipid species such as hexosylceramides and long-chain sphingomyelins, whose concentration physiologically increases in WT, and an increase in lipids typical of unspecialized plasma membranes, including phosphatidylcholines and phosphatidylethanolamines. Consistently, advanced morphometric analysis on more than 130,000 MFs revealed a delay in the evolution of CMT1A axon and myelin geometric parameters, appearing concomitantly with lipid impairment. We here demonstrate that, during normal development, MFs undergo a continuous maturation process in both chemical composition and physical structure, but these processes are delayed in CMT1A.
Erythropoiesis and Gene Expression Analysis in Erythroid Progenitor Cells Derived from Patients with Hemoglobin H/Constant Spring Disease
Authors: Wongkhammul Narawich , Khamphikham Pinyaphat , Tongjai Siripong , Tantiworawit Adisak , Fanhchaksai Kanda , Wongpalee Somsakul Pop , Tubsuwan Alisa , Maneekesorn Supawadee , Charoenkwan Pimlak
Published: October 2024

Hemoglobin H/Constant Spring (Hb H/CS) disease represents a form of non-deletional Hb H disease characterized by chronic hemolytic anemia that ranges from moderate to severe and may lead to transfusion-dependent thalassemia. To study the underlying mechanisms of this disease, we conducted an analysis of erythropoiesis and gene expression in erythroid progenitor cells derived from CD34+ hematopoietic stem/progenitor cells from patients with Hb H/CS disease and normal controls. Twelve patients with Hb H/CS disease and five normal controls were enrolled. Peripheral blood samples were collected to isolate CD34+ hematopoietic stem/progenitor cells for the analysis of cell proliferation and differentiation. Six samples from patients with Hb H/CS disease and three controls were subsequently studied for gene expression by next generation sequencing analysis. Erythroid progenitor cells derived from patients with Hb H/CS disease exhibited a trend towards increased rates of erythroid proliferation and decreased cell viability compared to those from controls. Moreover, erythroid progenitor cells derived from patients with Hb H/CS disease demonstrated delayed terminal differentiation. Gene expression profiling revealed elevated levels of genes encoding molecular chaperones, including the heat shock protein genes (HSPs) and the chaperonin containing TCP-1 subunit genes (CCTs) in the Hb H/CS disease group. In summary, erythroid progenitor cells derived from patients with Hb H/CS disease exhibit a trend towards heightened erythroid proliferation, diminished cell viability, and delayed terminal differentiation. Additionally, the increased expression of genes encoding molecular chaperones was observed, providing information on potential underlying pathophysiological mechanisms.
Encapsulation of Dexamethasone into mRNA–Lipid Nanoparticles Is a Promising Approach for the Development of Liver-Targeted Anti-Inflammatory Therapies
Authors: Rivero Berti Ignacio , Gambaro Rocío Celeste , Limeres María José , Huck-Iriart Cristián , Svensson Malin , Fraude-El Ghazi Silvia , Pretsch Leah , Si Shutian , Lieberwirth Ingo , Landfester Katharina , Cacicedo Maximiliano Luis , Islan Germán Abel , Gehring Stephan
Published: October 2024

The objective of this study was to develop two lipid nanoparticle (LNP) formulations capable of efficiently expressing a reporter mRNA while co-delivering the anti-inflammatory drug dexamethasone (DX) to reduce inflammatory side effects in protein replacement therapies. Two types of LNPs were developed, in which 25% of cholesterol was replaced by DX. These LNPs contained either 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) or 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) as a helper lipid. The resulting LNPs exhibited high stability, homogeneity, and near-neutral Zeta potentials. SAXS experiments confirmed DX incorporation into the LNP core, with slow in vitro DX release observed over 48 h. The LNPs achieved high mRNA encapsulation efficiency (95–100%) and effectively transfected HepG2 cells, dendritic cells, and hPBMCs. While LNPs increased cytokine release (IL-1β, TNF-α, MCP-1), LNPs-DX significantly reduced cytokine levels, demonstrating enhanced anti-inflammatory properties while maintaining mRNA expression levels. In vivo biodistribution showed predominant liver localization post-intramuscular injection, regardless of the DSPC or DOPE composition. LNPs co-loaded with mRNA and DX are promising candidates for continuous protein replacement. Due to their ability to reduce treatment-related inflammation while maintaining significant mRNA expression levels, these LNPs are perfectly suited for the treatment of liver-related metabolic diseases.
Unraveling the Pathogenesis of Calcinosis in Systemic Sclerosis: A Molecular and Clinical Insight
Authors: Avanoglu Guler Aslihan , De Luca Giacomo , Dagna Lorenzo , Matucci-Cerinic Marco , Campochiaro Corrado
Published: October 2024

Dystrophic calcinosis, which is the accumulation of insoluble calcified crystalline materials within tissues with normal circulating calcium and phosphorus levels, is a frequent finding in systemic sclerosis (SSc) and represents a major burden for patients. In SSc, calcinosis poses significant challenges in management due to the associated risk of severe complications such as infection, ulceration, pain, reduction in functional capacity and quality of life, and lack of standardized treatment choices. The exact pathogenesis of calcinosis is still unknown. There are multifaceted factors contributing to calcinosis development, including osteogenic differentiation of cells, imbalance between promoter and inhibitors of mineralization, local disturbance in calcium and phosphate levels, and extracellular matrix as a template for mineralization. Several pathophysiological changes observed in SSc such as ischemia, exacerbated production of excessive reactive oxygen species, inflammation, production of inflammatory cytokines, acroosteolysis, and increased extracellular matrix production may promote the development of calcinosis in SSc. Furthermore, mitochondrial dynamics, particularly fission function through the activity of dynamin-related protein-1, may have an effect on the dystrophic calcinosis process. In-depth investigations of cellular mechanisms and microenvironmental influences can offer valuable insights into the complex pathogenesis of calcinosis in SSc, providing potential targeting pathways for calcinosis treatment.
Masks of Albinism: Clinical Spectrum of Hermansky–Pudlak Syndrome
Authors: Bobreshova Anastasia M. , Ionova Sofya A. , Kadyshev Vitaly V. , Sukhanova Natella V. , Viakhireva Iuliia V. , Filatova Alexandra Yu. , Zhurkova Natalia V. , Sparber Peter A. , Marakhonov Andrey V. , Vasilyeva Tatyana A. , Shchagina Olga A. , Kutsev Sergey I. , Zinchenko Rena A.
Published: October 2024

Hermansky–Pudlak syndrome (HPS) is a rare disease inherited in the autosomal recessive mode, including 11 clinical genetic subtypes. They are associated with impaired function of the BLOC protein complex (Biogenesis of Lysosome-related Organelles Complexes), and the subunits of the AP-3 complex (adaptor protein complex). Each has its own clinical features, but they are all characterized by albinism, bleeding disorder, and visual abnormalities. Eleven patients from eight unrelated families with an incoming diagnosis of albinism were examined and novel and previously described genetic variants in HPS1, HPS6, and BLOC1S6 genes (types HPS1, HPS6, and HPS9) were found. To determine the optimal therapy and recommendations for further follow up, it is necessary to consider the entire clinical spectrum and genetic polymorphism of the disease. An interdisciplinary approach, combined with the use of non-routine diagnostic techniques such as RNA analysis, is essential for achieving accurate diagnoses in certain complex cases.
Pulmonary Congestion and Anemia in Hemodialysis: The Potential Link to Inflammation
Authors: Kaysi Saleh , Pacha Bakhtar , Antoine Marie-Hélène , De Prez Eric , Nortier Joëlle
Published: October 2024

Pulmonary congestion (PC) is common in hemodialysis (HD) patients. We explored the association of anemia and pulmonary congestion in HD patients. A prospective pilot observational study included 18 patients on maintenance HD. Individual B-lines scores (BLS; 8-sites method) were obtained by lung ultrasound, before and after the first two consecutive HD sessions of the week (HD1-HD2), with different inter-dialytic intervals (68 vs. 44 h). Bioimpedance spectroscopy body composition (BIS) was performed before each HD session. Hemoglobin (Hb) levels, in addition to circulating markers of chronic inflammation (soluble urokinase Plasminogen Activator Receptor [suPAR], soluble Suppression of Tumorigenicity 2 [sST2]) were obtained. Mean (±SD) BLS values were quite elevated at all time points: Pre-HD1 (16 ± 5.53), post-HD1 (15.3 ± 6.63), pre-HD2 (16.3 ± 5.26) and post-HD2 (13.6 ± 5.83), respectively. No direct significant correlation was found between inflammation markers levels and BLS. However, mean levels (±SD, ng/mL) of suPAR pre-HD1 (7.88 ± 3.07) and pre-HD2 (7.78 ± 3.02) remained significantly above the normal range (<4 ng/mL), and sST2 levels reached 2-fold the upper normal value in most patients (27.4 ± 17.8). Pulmonary congestion reflected by BLS was negatively correlated to Hb levels pre-HD1 (R² = 0.439, p = 0.003), and pre-HD2 (R² = 0.301, p = 0.018). In addition, Hb levels were negatively correlated to global volume status estimated by BIS (R² = 0.351, p = 0.009). Hemoglobin levels were negatively correlated to pulmonary congestion and to the global volume status evaluated by BIS. Chronic inflammation markers were increased in HD patients, suggesting a complex volume- and non-volume-dependent pathophysiology of pulmonary congestion in HD patients.