bluuebunny/crossref_metadata_2025 huggingface dataset:
Identification and Prediction of Differentially Expressed MicroRNAs Associated with Detoxification Pathways in Larvae of Spodoptera frugiperda
Authors: Wang Yan-Ping , Chen Xing-Yu , Pu De-Qiang , Yi Chun-Yan , Liu Chang-Hua , Zhang Cui-Cui , Wei Zhen-Zhen , Guo Jing-Wei , Yu Wen-Juan , Chen Song , Liu Hong-Ling
Published: August 2024

Spodoptera frugiperda poses a severe threat to crops, causing substantial economic losses. The increased use of chemical pesticides has led to resistance in S. frugiperda populations. Micro ribonucleic acids (MicroRNAs or miRNAs) are pivotal in insect growth and development. This study aims to identify miRNAs across different developmental stages of S. frugiperda to explore differential expression and predict target gene functions. High-throughput sequencing of miRNAs was conducted on eggs, 3rd instar larvae, pupae, and adults. Bioinformatics analyses identified differentially expressed miRNAs specifically in larvae, with candidate miRNAs screened to predict target genes, particularly those involved in detoxification pathways. A total of 184 known miRNAs and 209 novel miRNAs were identified across stages. Comparative analysis revealed 54, 15, and 18 miRNAs differentially expressed in larvae, compared to egg, pupa, and adult stages, respectively. Eight miRNAs showed significant differential expression across stages, validated by quantitative reverse transcription PCR (qRT-PCR). Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses predicted target genes’ functions, identifying eight differentially expressed miRNAs targeting 10 gene families associated with detoxification metabolism, including P450s, glutathione S-transferase (GSTs), ATP-binding cassette (ABC) transporters, and sodium channels. These findings elucidate the species-specific miRNA profiles and regulatory mechanisms of detoxification-related genes in S. frugiperda larvae, offering insights and strategies for effectively managing this pest.
Aberrantly Expressed tRNA-Val Fragments Can Distinguish Canine Hepatocellular Carcinoma from Canine Hepatocellular Adenoma
Authors: Hashimoto Saki , Hasan MD , Arif Mohammad , Nozaki Nobuhiro , Husna Al , Furusawa Yu , Sogawa Takeshi , Takahashi Kaori , Kuramoto Tomohide , Noguchi Aki , Takahashi Masashi , Yamato Osamu , Rahman Md , Miura Naoki
Published: August 2024

Hepatocellular adenoma (HCA) and hepatocellular carcinoma (HCC) can be difficult to differentiate but must be diagnosed correctly as treatment and prognosis for these tumors differ markedly. Relevant diagnostic biomarkers are thus needed, and those identified in dogs may have utility in human medicine because of the similarities between human and canine HCA and HCC. A tRNA-derived fragment (tRF), tRNA-Val, is a promising potential biomarker for canine mammary gland tumors but has not previously been investigated in hepatic tumors. Accordingly, we aimed to elucidate the potential utility of tRNA-Val as a biomarker for canine HCA and HCC using clinical samples (tumor tissue and plasma extracellular vesicles [EVs]) and tumor cell lines with qRT-PCR assays. We also investigated relevant functions and signaling pathways with bioinformatic analyses (Gene Ontology and Kyoto Encyclopedia of Genes and Genomes). tRNA-Val was markedly downregulated in HCC tumor tissue versus HCA tumor tissue and normal liver tissue, and a similar trend was shown in plasma EVs and HCC cell lines versus healthy controls. Based on areas under the receiver operating characteristic curves (AUCs), tRNA-Val significantly distinguished HCC (AUC = 1.00, p = 0.001) from healthy controls in plasma EVs and HCC from HCA (AUC = 0.950, p = 0.01). Bioinformatics analysis revealed that tRNA-Val may be primarily involved in DNA repair, mRNA processing, and splicing and may be linked to the N-glycan and ubiquitin-mediated proteasome pathways. This is the first report on the expression of tRNA-Val in canine HCC and HCA and its possible functions and signaling pathways. We suggest that tRNA-Val could be a promising novel biomarker to distinguish canine HCC from HCA. This study provides evidence for a greater understanding of the role played by tRNA-Val in the development of canine HCC.
The First FISH-Confirmed Non-Canonical Telomeric Motif in Heteroptera: Cimex lectularius Linnaeus, 1758 and C. hemipterus (Fabricius, 1803) (Hemiptera, Cimicidae) Have a 10 bp Motif (TTAGGGATGG)n
Authors: Stoianova Desislava , Grozeva Snejana , Golub Natalia V. , Anokhin Boris A. , Kuznetsova Valentina G.
Published: August 2024

Fluorescence in situ hybridization (FISH) with two different probes, the canonical insect telomeric sequence (TTAGG)n and the sequence (TTAGGGATGG)n, was performed on meiotic chromosomes of two members of the true bug family Cimicidae (Cimicomorpha), the common bed bug Cimex lectularius Linnaeus, 1758 and the tropical bed bug C. hemipterus (Fabricius, 1803), whose telomeric motifs were not known. In both species, there were no hybridization signals with the first probe, but strong signals at chromosomal ends were observed with the second probe, indicating the presence of a telomeric motif (TTAGGGATGG)n. This study represents the first FISH confirmation of the presence of a non-canonical telomeric motif not only for the infraorder Cimicomorpha but also for the suborder Heteroptera (Hemiptera) as a whole. The present finding is of key significance for unraveling the evolutionary shifts in the telomeric sequences in this suborder.
Knowledge, Acceptance, and Uptake of Family Planning: A Cluster Randomized Controlled Trial of Group Antenatal Care in Ghana
Authors: Zielinski Ruth , Abdelnabi Samia , Amankwah Georgina , Kukula Vida A. , Apetorgbor Veronica , Awini Elizabeth , Williams John , Moyer Cheryl , Ghosh Bidisha , Lori Jody R.
Published: August 2024

The use of family planning (FP) methods significantly contributes to improved outcomes for mothers and their offspring. However, the use of FP remains low, particularly in low- and middle-income countries. A cluster randomized controlled clinical trial was implemented in Ghana, comparing group antenatal care (ANC) with routine care. The group ANC intervention included eight meetings where the seventh group meeting incorporated information and discussion regarding methods of FP. Data collection occurred at five time points: baseline (T0), 34 weeks’ gestation (T1), 6–12 weeks post birth (T2), 5–8 months post birth, and 11–14 months post birth (T4). At T1, there was a significantly greater increase in the knowledge of FP methods as well as the intention to use FP after the birth among the intervention group. The uptake of FP was significantly higher in the intervention group for all post-birth timepoints except for T4 where the control group had significantly higher rates. The reasons for the diminishing effect are unclear. An increasing uptake of FP methods requires a multifaceted approach that includes increasing accessibility, knowledge, and acceptability as well as addressing societal and cultural norms.
Bi-Directional and Time-Lagged Associations between Engagement and Mental Health Symptoms in a Group Mindfulness-Based Mental Health Intervention
Authors: Rigsby Brock A. , Miller Reagan L. , Moran Megan J. , Rzonca Addie J. , Najman Jonathan I. , Adams Melanie S. , Prince Mark A. , Lucas-Thompson Rachel G.
Published: August 2024

There is a high need for accessible avenues for improving mental health among emerging adults, particularly on college campuses. Mindfulness-based intervention (MBI) is a promising avenue for reducing mental health symptoms, but initial discomforts associated with MBI may cause symptoms to fluctuate before decreasing, which presents a barrier to engagement with mindfulness on a daily basis. Consistent mindfulness practice is key for forming habits related to MBI, and engagement with mindfulness at home, including between intervention sessions, is an important predictor of mental health outcomes. Research suggests that mental health symptoms may serve as barriers to their own treatment. Thus, it is important to understand how mental health symptom levels impact adherence to treatment protocols. To improve understanding of symptom-specific barriers to treatment and engagement with mindfulness, the present study collected daily diary surveys about engagement with mindfulness and mental health symptoms from a sample of 62 adults recruited to participate in a six-week mindfulness intervention. We explored mental health symptoms as a predictor of engagement with MBI at the mean level and whether within-person variability in symptoms predicted same-day or time-lagged changes in engagement via mixed-effects associations. Using heterogeneous location scale models, we further explored whether erraticism in either mental health symptoms or engagement with mindfulness predicted the other and if outcomes of the mindfulness intervention were homogeneous among subjects. Results showed that bi-directional and time-lagged associations exist between symptoms and engagement, indicating that there is a nuanced temporal and reciprocal relationship between engagement with mindfulness and mental health symptoms. Daily within-person elevations in engagement with mindfulness were associated with concurrent improvements in mental health but prospective increases in mental health symptoms. We also found that higher engagement (over personal averages) was not consistently associated with improvements in mental health across the sample but was instead associated with greater heterogeneity in outcomes. We also found that increases in mental health symptoms (over personal averages), as well as higher average levels of mental health symptoms, were both associated with lower levels of engagement in the mindfulness treatment protocol.
Patients with Thyroid Dyshormonogenesis and DUOX2 Variants: Molecular and Clinical Description and Genotype–Phenotype Correlation
Authors: Baz-Redón Noelia , Antolín María , Clemente María , Campos Ariadna , Mogas Eduard , Fernández-Cancio Mónica , Zafon Elisenda , García-Arumí Elena , Soler Laura , González-Llorens Núria , Aguilar-Riera Cristina , Camats-Tarruella Núria , Yeste Diego
Published: August 2024

Thyroid dyshormonogenesis (THD) is a heterogeneous group of genetic diseases caused by the total or partial defect in the synthesis or secretion of thyroid hormones. Genetic variants in DUOX2 can cause partial to total iodination organification defects and clinical heterogeneity, from transient to permanent congenital hypothyroidism. The aim of this study was to undertake a molecular characterization and genotype–phenotype correlation in patients with THD and candidate variants in DUOX2. A total of 31 (19.38%) patients from the Catalan Neonatal Screening Program presented with variants in DUOX2 that could explain their phenotype. Fifteen (48.39%) patients were compound heterozygous, 10 (32.26%) heterozygous, and 4 (12.90%) homozygous. In addition, 8 (26.67%) of these patients presented variants in other genes. A total of 35 variants were described, 10 (28.57%) of these variants have not been previously reported in literature. The most frequent variant in our cohort was c.2895_2898del/p.(Phe966SerfsTer29), classified as pathogenic according to reported functional studies. The final diagnosis of this cohort was permanent THD in 21 patients and transient THD in 10, according to reevaluation and/or need for treatment with levothyroxine. A clear genotype–phenotype correlation could not be identified; therefore, functional studies are necessary to confirm the pathogenicity of the variants.
The Interrelationships between Cytokines and Schizophrenia: A Systematic Review
Authors: Lv Haibing , Guo Meng , Guo Chuang , He Kuanjun
Published: August 2024

Schizophrenia (SCZ) imposes a significant burden on patients and their families because of its high prevalence rate and disabling nature. Given the lack of definitive conclusions regarding its pathogenesis, physicians heavily rely on patients’ subjective symptom descriptions for diagnosis because reliable diagnostic biomarkers are currently unavailable. The role of the inflammatory response in the pathogenesis of SCZ has been supported by some studies. The findings of these studies showed abnormal changes in the levels of inflammatory factors, such as cytokines (CKs), in both peripheral blood and cerebrospinal fluid (CSF) among individuals affected by SCZ. The findings imply that inflammatory factors could potentially function as risk indicators for the onset of SCZ. Consequently, researchers have directed their attention towards investigating the potential utility of CKs as viable biomarkers for diagnosing SCZ. Extracellular vesicles (EVs) containing disease-specific components exhibit remarkable stability and abundance, making them promising candidates for biomarker discovery across various diseases. CKs encapsulated within EVs secreted by immune cells offer valuable insights into disease progression. This review presents a comprehensive analysis summarizing the relationship between CKs and SCZ and emphasizes the vital role of CKs encapsulated within EVs in the pathogenesis and development of SCZ.
COVID-19-Associated Sepsis: Potential Role of Phytochemicals as Functional Foods and Nutraceuticals
Authors: de Souza Goncalves Bruno , Sangani Darshan , Nayyar Aleen , Puri Raghav , Irtiza Mahir , Nayyar Asma , Khalyfa Abdelnaby , Sodhi Komal , Pillai Sneha S.
Published: August 2024

The acute manifestations of coronavirus disease 2019 (COVID-19) exhibit the hallmarks of sepsis-associated complications that reflect multiple organ failure. The inflammatory cytokine storm accompanied by an imbalance in the pro-inflammatory and anti-inflammatory host response to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection leads to severe and critical septic shock. The sepsis signature in severely afflicted COVID-19 patients includes cellular reprogramming and organ dysfunction that leads to high mortality rates, emphasizing the importance of improved clinical care and advanced therapeutic interventions for sepsis associated with COVID-19. Phytochemicals of functional foods and nutraceutical importance have an incredible impact on the healthcare system, which includes the prevention and/or treatment of chronic diseases. Hence, in the present review, we aim to explore the pathogenesis of sepsis associated with COVID-19 that disrupts the physiological homeostasis of the body, resulting in severe organ damage. Furthermore, we have summarized the diverse pharmacological properties of some potent phytochemicals, which can be used as functional foods as well as nutraceuticals against sepsis-associated complications of SARS-CoV-2 infection. The phytochemicals explored in this article include quercetin, curcumin, luteolin, apigenin, resveratrol, and naringenin, which are the major phytoconstituents of our daily food intake. We have compiled the findings from various studies, including clinical trials in humans, to explore more into the therapeutic potential of each phytochemical against sepsis and COVID-19, which highlights their possible importance in sepsis-associated COVID-19 pathogenesis. We conclude that our review will open a new research avenue for exploring phytochemical-derived therapeutic agents for preventing or treating the life-threatening complications of sepsis associated with COVID-19.
Molecular Pro-Apoptotic Activities of Flavanone Derivatives in Cyclodextrin Complexes: New Implications for Anticancer Therapy
Authors: Adamus-Grabicka Angelika A. , Hikisz Pawel , Stepniak Artur , Malecka Magdalena , Paneth Piotr , Sikora Joanna , Budzisz Elzbieta
Published: August 2024

This study evaluates the antiproliferative potential of flavanones, chromanones and their spiro-1-pyrazoline derivatives as well as their inclusion complexes. The main goal was to determine the biological basis of molecular pro-apoptotic activities and the participation of reactive oxygen species (ROS) in shaping the cytotoxic properties of the tested conjugates. For this purpose, changes in mitochondrial potential and the necrotic/apoptotic cell fraction were analyzed. Testing with specific fluorescent probes found that ROS generation had a significant contribution to the biological anticancer activity of complexes of flavanone analogues. TT (thrombin time), PT (prothrombin time) and APTT (activated partial tromboplastin time) were used to evaluate the influence of the compounds on the extrinsic and intrinsic coagulation pathway. Hemolysis assays and microscopy studies were conducted to determine the effect of the compounds on RBCs.
Origins and Genetic Characteristics of Egyptian Peach
Authors: Ezzat Mohamed , Zhang Weihan , Amar Mohamed , Nishawy Elsayed , Zhao Lei , Belal Mohammad , Han Yuepeng , Liao Liao
Published: August 2024

Peach (Prunus persica), a significant economic fruit tree in the Rosaceae family, is extensively cultivated in temperate and subtropical regions due to its abundant genetic diversity, robust adaptability, and high nutritional value. Originating from China over 4000 years ago, peaches were introduced to Persia through the Silk Road during the Han Dynasty and gradually spread to India, Greece, Rome, Egypt, Europe, and America. Currently grown in more than 80 countries worldwide, the expansion of peach cultivation in Egypt is mainly due to the development and utilization of peach varieties with low chilling requirements. These varieties exhibit unique phenotypic characteristics such as early maturity, reduced need for winter cold temperatures, low water requirements, and high economic value. In this study, a systematic analysis was conducted on the genetic characteristics and kinship relationships of peaches with low chilling requirements in Egypt. We conducted a comprehensive evolutionary and Identity-by-Descent (IBD) analysis on over 300 peach core germplasm resources, including Egyptian cultivars with low chilling requirements, to investigate their origin and genetic characteristics. The evolutionary analysis revealed that ‘Bitter almond’ is closely related to China’s wild relative species Prunus tangutica Batal, while ‘Early grand’ shares one branch with Chinese ornamental peach cultivars, and ‘Nemaguard’ clusters with some ancient local varieties from China. The IBD analysis also indicated similar genetic backgrounds, suggesting a plausible origin from China. Similarly, the analysis suggested that ‘Swelling’ may have originated from the Czech Republic while ‘Met ghamr’ has connections to South Africa. ‘Desert red’, ‘Early swelling’, and ‘Florida prince’ are likely derived from Brazil. These findings provide valuable insights into the genetic characteristics of Egyptian peach cultivars. They offer a significant foundation for investigating the origin and spread of cultivated peaches worldwide and serve as a valuable genetic resource for breeding low chilling requirement cultivars, which is of considerable significance for the advancement of peach cultivation in Egypt.
Molecular and Pathological Features of Paediatric High-Grade Gliomas
Authors: Blasco-Santana Luis , Colmenero Isabel
Published: August 2024

Paediatric high-grade gliomas are among the most common malignancies found in children. Despite morphological similarities to their adult counterparts, there are profound biological and molecular differences. Furthermore, and thanks to molecular biology, the diagnostic pathology of paediatric high-grade gliomas has experimented a dramatic shift towards molecular classification, with important prognostic implications, as is appropriately reflected in both the current WHO Classification of Tumours of the Central Nervous System and the WHO Classification of Paediatric Tumours. Emphasis is placed on histone 3, IDH1, and IDH2 alterations, and on Receptor of Tyrosine Kinase fusions. In this review we present the current diagnostic categories from the diagnostic pathology perspective including molecular features.
Genome-Wide Identification of the Brassinosteroid Signal Kinase Gene Family and Its Profiling under Salinity Stress
Authors: Shi Biao , Wang Youwu , Wang Liang , Zhu Shengwei
Published: August 2024

Alfalfa (Medicago L.) is a high-quality perennial leguminous forage with the advantages of salt tolerance, mowing tolerance, high protein content, and other economically valuable characteristics. As the sixth class of plant hormones, brassinosteroids (BRs) play indispensable roles in modulating a variety of plant growth, maturation, and environmental adaptation processes, thereby influencing vegetal expansion and development. Brassinosteroid signal kinases (BSKs) are key cytoplasmic receptor kinases downstream of the BR signaling transduction pathway, participating in plant growth, development, and stress regulation. However, the phylogenetic and expression pattern analyses of the BSK gene family among the five alfalfa species have rarely been reported; in this study, 52 BSK family members were found in the genomes of the five subspecies, and phylogenetic trees were constructed according to protein sequences, allowing us to categorize all BSKs into seven distinct groups. Domain, conserved motif, and exon–intron structural analyses showed that most BSK members were relatively conserved, except for MtBSK3-2, MtBSK7-1, and MtBSK7-2, which may be truncated members. Intra-species collinearity and Ka/Ks analyses showed that purifying selection influenced BSK genes during evolution; most of the cis-acting elements in the promoter region were associated with responses, such as light, defense, and stress, anaerobic induction, MeJA, and abscisic acid. Expression pattern analysis indicated that the majority of alfalfa genes exhibited downregulation after reaching a peak at 0.5 h after treatment with 250 mM NaCl, especially for MsBSK14, MsBSK15, MsBSK17, MsBSK19, and MsBSK21; meanwhile, MsBSK4, MsBSK7, and MsBSK9 increased and were highly expressed at 12 h, demonstrating significantly altered expression patterns under salt stress; furthermore, MsBSK4, MsBSK7, and MsBSK9 exhibited expression specifically in the leaves. qRT-PCR analysis confirmed the expression trends for MsBSK4, MsBSK7, MsBSK9, MsBSK14, MsBSK15, and MsBSK16 matched the transcriptome data. However, the trends for MsBSK17, MsBSK19, and MsBSK21 diverged from the transcriptome data. Our study may provide a foundation for further functional analyses of BSK genes in growth, development, and salt stress tolerance in alfalfa.
Epigenetic Regulation and Molecular Mechanisms of Burn Injury-Induced Nociception in the Spinal Cord of Mice
Authors: Mészár Zoltán , Erdei Virág , Szücs Péter , Varga Angelika
Published: August 2024

Epigenetic mechanisms, including histone post-translational modifications (PTMs), play a critical role in regulating pain perception and the pathophysiology of burn injury. However, the epigenetic regulation and molecular mechanisms underlying burn injury-induced pain remain insufficiently explored. Spinal dynorphinergic (Pdyn) neurons contribute to heat hyperalgesia induced by severe scalding-type burn injury through p-S10H3-dependent signaling. Beyond p-S10H3, burn injury may impact various other histone H3 PTMs. Double immunofluorescent staining and histone H3 protein analyses demonstrated significant hypermethylation at H3K4me1 and H3K4me3 sites and hyperphosphorylation at S10H3 within the spinal cord. By analyzing Pdyn neurons in the spinal dorsal horn, we found evidence of chromatin activation with a significant elevation in p-S10H3 immunoreactivity. We used RNA-seq analysis to compare the effects of burn injury and formalin-induced inflammatory pain on spinal cord transcriptomic profiles. We identified 98 DEGs for burn injury and 86 DEGs for formalin-induced inflammatory pain. A limited number of shared differentially expressed genes (DEGs) suggest distinct central pain processing mechanisms between burn injury and formalin models. KEGG pathway analysis supported this divergence, with burn injury activating Wnt signaling. This study enhances our understanding of burn injury mechanisms and uncovers converging and diverging pathways in pain models with different origins.
Optimization of the Method for Isolating Bacterial DNA from the Aboveground Part of Lettuce
Authors: Krupka Magdalena , Piotrowicz-Cieślak Agnieszka I.
Published: August 2024

Developing an effective method for isolating bacterial genetic material from plants is a relatively challenging task and often does not yield adequately prepared material for further analyses. Previous studies often overlook connections, primarily focusing on laboratory investigations. With advancements in high-throughput sequencing techniques, we can now revisit and delve deeper into these interactions. Our study focuses on the initial phase of these investigations: genetic material isolation. Extracting bacterial DNA from aboveground plant parts, known as the phyllosphere, poses a significant challenge due to plant-derived contaminants. Existing isolation protocols frequently yield inconsistent results, necessitating continuous refinement and optimization. In our study, we developed an effective isolation protocol employing mechanical–chemical lysis, sonication, and membrane filtration. This approach yielded high-quality DNA at a concentration of 38.08 ng/µL, suitable for advanced sequencing applications. Our results underscore the effectiveness and necessity of these methods for conducting comprehensive microbiological analyses. Furthermore, our research not only lays the groundwork for further studies on lettuce microbiota, but also highlights the potential for utilizing our developed protocol in investigating other plants and their microbiomes.
Multi-Level Protocol for Mechanistic Reaction Studies Using Semi-Local Fitted Potential Energy Surfaces
Authors: Piskor Tomislav , Pinski Peter , Mast Thilo , Rybkin Vladimir
Published: August 2024

In this work, we propose a multi-level protocol for routine theoretical studies of chemical reaction mechanisms. The initial reaction paths of our investigated systems are sampled using the Nudged Elastic Band (NEB) method driven by a cheap electronic structure method. Forces recalculated at the more accurate electronic structure theory for a set of points on the path are fitted with a machine learning technique (in our case symmetric gradient domain machine learning or sGDML) to produce a semi-local reactive potential energy surface (PES), embracing reactants, products and transition state (TS) regions. This approach has been successfully applied to a unimolecular (Bergman cyclization of enediyne) and a bimolecular (SN2 substitution) reaction. In particular, we demonstrate that with only 50 to 150 energy-force evaluations with the accurate reference methods (here complete-active-space self-consistent field, CASSCF, and coupled-cluster singles and doubles, CCSD) it is possible to construct a semi-local PES giving qualitative agreement for stationary-point geometries, intrinsic reaction coordinates and barriers. Furthermore, we find a qualitative agreement in vibrational frequencies and reaction rate coefficients. The key aspect of the method’s performance is its multi-level nature, which not only saves computational effort but also allows extracting meaningful information along the reaction path, characterized by zero gradients in all but one direction. Agnostic to the nature of the TS and computationally economic, the protocol can be readily automated and routinely used for mechanistic reaction studies.
Accurate and Automated Genotyping of the CFTR Poly-T/TG Tract with CFTR-TIPS
Authors: Ding Qiliang , Hofich Christopher D. , Kellogg Tifani B. , Kuennen Rhonda K. , Paxton Kaitlin N. , Thieke Sarah M. , Rumilla Kandelaria M. , Hasadsri Linda
Published: August 2024

Cystic fibrosis is caused by biallelic pathogenic variants in the CFTR gene, which contains a polymorphic (TG)mTn sequence (the “poly-T/TG tract”) in intron 9. While T9 and T7 alleles are benign, T5 alleles with longer TG repeats, e.g., (TG)12T5 and (TG)13T5, are clinically significant. Thus, professional medical societies currently recommend reporting the TG repeat size when T5 is detected. Sanger sequencing is a cost-effective method of genotyping the (TG)mTn tract; however, its polymorphic length substantially complicates data analysis. We developed CFTR-TIPS, a freely available web-based software tool that infers the (TG)mTn genotype from Sanger sequencing data. This tool detects the (TG)mTn tract in the chromatograms, quantifies goodness of fit with expected patterns, and visualizes the results in a graphical user interface. It is broadly compatible with any Sanger chromatogram that contains the (TG)mTn tract ± 15 bp. We evaluated CFTR-TIPS using 835 clinical samples previously analyzed in a CLIA-certified, CAP-accredited laboratory. When operated fully automatically, CFTR-TIPS achieved 99.8% concordance with our clinically validated manual workflow, while generally taking less than 10 s per sample. There were two discordant samples: one due to a co-occurring heterozygous duplication that confounded the tool and the other due to incomplete (TG)mTn tract detection in the reverse chromatogram. No clinically significant misclassifications were observed. CFTR-TIPS is a free, accurate, and rapid tool for CFTR (TG)mTn tract genotyping using cost-effective Sanger sequencing. This tool is suitable both for automated use and as an aid to manual review to enhance accuracy and reduce analysis time.
Analysis of Amyloid Fibrillation of Two Family 1 Glycoside Hydrolases
Authors: Rodríguez-López Miguel A. , Coll-Marqués José María , Talens-Perales David , Marín-Navarro Julia , Polaina Julio , Vázquez-Contreras Edgar
Published: August 2024

The formation and analysis of amyloid fibers by two β-glucosidases, BglA and BglB, belonging to the GH1 enzyme family, are reported. Both proteins have the (β/α)8 TIM-barrel fold, which is characteristic of this family and is also the most common protein structure. BglA is an octamer, whereas BglB is a monomer. Amyloid fibrillation using pH and temperature as perturbing agents was investigated using fluorescence spectroscopy as a preliminary approach and corroborated using wide-field optical microscopy, confocal microscopy, and field-emission scanning electron microscopy. These analyses showed that both enzymes fibrillate at a wide range of acidic and alkaline conditions and at several temperature conditions, particularly at acidic pH (3–4) and at temperatures between 45 and 65 °C. Circular dichroism spectroscopy corroborated the transition from an α-helix to a β-sheet secondary structure of both proteins in conditions where fibrillation was observed. Overall, our results suggest that fibrillation is a rather common phenomenon caused by protein misfolding, driven by a transition from an α-helix to a β-sheet secondary structure, that many proteins can undergo if subjected to conditions that disturb their native conformation.
Application and Development of Cell Membrane Functionalized Biomimetic Nanoparticles in the Treatment of Acute Ischemic Stroke
Authors: Li Ying , Wu Chuang , Yang Rui , Tang Jiannan , Li Zhanqing , Yi Xue , Fan Zhongxiong
Published: August 2024

Ischemic stroke is a serious neurological disease involving multiple complex physiological processes, including vascular obstruction, brain tissue ischemia, impaired energy metabolism, cell death, impaired ion pump function, and inflammatory response. In recent years, there has been significant interest in cell membrane-functionalized biomimetic nanoparticles as a novel therapeutic approach. This review comprehensively explores the mechanisms and importance of using these nanoparticles to treat acute ischemic stroke with a special emphasis on their potential for actively targeting therapies through cell membranes. We provide an overview of the pathophysiology of ischemic stroke and present advances in the study of biomimetic nanoparticles, emphasizing their potential for drug delivery and precision-targeted therapy. This paper focuses on bio-nanoparticles encapsulated in bionic cell membranes to target ischemic stroke treatment. It highlights the mechanism of action and research progress regarding different types of cell membrane-functionalized bi-onic nanoparticles such as erythrocytes, neutrophils, platelets, exosomes, macrophages, and neural stem cells in treating ischemic stroke while emphasizing their potential to improve brain tissue’s ischemic state and attenuate neurological damage and dysfunction. Through an in-depth exploration of the potential benefits provided by cell membrane-functionalized biomimetic nanoparticles to improve brain tissue’s ischemic state while reducing neurological injury and dysfunction, this study also provides comprehensive research on neural stem cells’ potential along with that of cell membrane-functionalized biomimetic nanoparticles to ameliorate neurological injury and dysfunction. However, it is undeniable that there are still some challenges and limitations in terms of biocompatibility, safety, and practical applications for clinical translation.
Therapeutic Transplantation of Human Central Nervous System Organoids for Neural Reconstruction
Authors: Hong Sung Jun , Bock Minsung , Zhang Songzi , An Seong Bae , Han Inbo
Published: August 2024

Damage to the central nervous system (CNS) often leads to irreversible neurological deficits, and there are currently few effective treatments available. However, recent advancements in regenerative medicine have identified CNS organoids as promising therapeutic options for addressing CNS injuries. These organoids, composed of various neurons and supporting cells, have shown potential for direct repair at injury sites. CNS organoids resemble the structure and function of actual brain tissue, which allows them to adapt and function well within the physiological environment when transplanted into injury sites. Research findings suggest that CNS organoids can replace damaged neurons, form new neural connections, and promote neural recovery. This review highlights the emerging benefits, evaluates preclinical transplantation outcomes, and explores future strategies for optimizing neuroregeneration using CNS organoids. With continued research and technological advancements, these organoids could provide new hope for patients suffering from neurological deficits.
Evaluating the Efficiency of zk-SNARK, zk-STARK, and Bulletproof in Real-World Scenarios: A Benchmark Study
Authors: El-Hajj Mohammed , Oude Roelink Bjorn
Published: August 2024

This study builds on our previous systematic literature review (SLR) that assessed the applications and performance of zk-SNARK, zk-STARK, and Bulletproof non-interactive zero-knowledge proof (NIZKP) protocols. To address the identified research gaps, we designed and implemented a benchmark comparing these three protocols using a dynamic minimized multiplicative complexity (MiMC) hash application. We evaluated performance across four general-purpose programming libraries and two programming languages. Our results show that zk-SNARK produced the smallest proofs, while zk-STARK generated the largest. In terms of proof generation and verification times, zk-STARK was the fastest, and Bulletproof was the slowest. Interestingly, zk-SNARK proofs verified marginally faster than zk-STARK, contrary to other findings. These insights enhance our understanding of the functionality, security, and performance of NIZKP protocols, providing valuable guidance for selecting the most suitable protocol for specific applications.
Structure and Evolution of Ribosomal Genes of Insect Chromosomes
Authors: Gokhman Vladimir E. , Kuznetsova Valentina G.
Published: August 2024

Currently, clusters of 45S and 5S ribosomal DNA (rDNA) have been studied in about 1000 and 100 species of the class Insecta, respectively. Although the number of insect species with known 45S rDNA clusters (also referred to as nucleolus-organizing regions, or NORs) constitutes less than 0.1 percent of the described members of this enormous group, certain conclusions can already be drawn. Since haploid karyotypes with single 45S and 5S rDNA clusters predominate in both basal and derived insect groups, this character state is apparently ancestral for the class Insecta in general. Nevertheless, the number, chromosomal location, and other characteristics of both 45S and 5S rDNA sites substantially vary across different species, and sometimes even within the same species. There are several main factors and molecular mechanisms that either maintain these parameters or alter them on the short-term and/or long-term scale. Chromosome structure (i.e., monocentric vs. holokinetic chromosomes), excessive numbers of rRNA gene copies per cluster, interactions with transposable elements, pseudogenization, and meiotic recombination are perhaps the most important among them.
Noninvasive Mapping System for the Stereotactic Radioablation Treatment of Ventricular Tachycardia: A Case Description
Authors: Romanazzi Imma , Di Monaco Antonio , Bonaparte Ilaria , Valenti Noemi , Surgo Alessia , Di Guglielmo Fiorella , Fiorentino Alba , Grimaldi Massimo
Published: August 2024

Objectives: Sustained monomorphic ventricular tachycardia (SMVT) is a life-threatening condition that is often observed in patients with structural heart disease. Catheter ablation (CA) ablation is an effective and well-established treatment for the scar-related ventricular tachycardias (VTs). Sometimes, due to patient fragility or contraindications to CA, a noninvasive procedure is required. In these cases, VT ablation with stereotactic arrhythmia radioablation (STAR) for SMVTs supported by the CardioInsight mapping system seems to be a promising and effective noninvasive approach. Methods and results: We report a case of a 55-year-old male smoker and heavy alcohol consumer who developed ischemic heart disease and frequent refractory SMVT relative to antiarrhythmic drugs. Catheter ablation was not practicable due to the presence of an apical thrombosis in the left ventricle. The CardioInsightTM system (Cardioinsight Technologies Inc., Cleveland, OH, USA) was useful for noninvasively mapping the VTs, identifying two target areas on the septum and anterior wall of the left ventricle. A personalized STAR treatment plan was carefully designed, and it was delivered in a few minutes. During follow-up, a significant reduction in the arrhythmia burden was documented. Conclusions: Stereotactic arrhythmia radioablation supported by the CardioInsight system could be an alternative treatment for VTs when catheter ablation is not possible. Larger studies are needed to investigate this technique.
Monoclonal Anti-PCSK9 Antibodies: Real-World Data
Authors: Guidotti Giulia , Liberati Viola , Sorrentino Andrea , Lotti Elena , Crudele Felice , Rogolino Angela , Sammartino Aniello , Slanzi Margherita , Gori Anna Maria , Marcucci Rossella , Berteotti Martina
Published: August 2024

Background: Real-world data on the use of lipid-lowering therapy (LLT) in clinical practice show that about 80% of (very) high-cardiovascular (CV)-risk patients disregard the 2019 European Society of Cardiology (ESC) Guidelines’ recommendations on dyslipidemias. The availability of proprotein convertase subtilisin/kexin type 9 monoclonal antibodies (PCSK9mAb) should reduce this gap. Our aim was to provide data on PCSK9mAb use in clinical practice, investigating the achievement of the ESC Guidelines’ recommendations in the real world. Methods: Between April 2018 and December 2022, patients who started on PCSK9mAb therapy (140 mg of evolocumab or 75 mg or 150 mg of alirocumab, subcutaneous injection every 2 weeks) were included in a prospective registry. Our cohort consisted of 256 patients: 95 (37.1%) were women (mean age: 65.43 ± 11.12 yrs), 53 (20.7%) were at high CV risk, and 203 (79.3%) were at very high CV risk. Results: After one year of PCSK9mAb treatment, nearly 60% of patients demonstrated full adherence to the ESC Guidelines’ recommendations, defined as achieving at least a 50% reduction in low-density lipoprotein cholesterol (LDL-C) levels along with reaching LDL-C target levels (≤55 and ≤70 mg/dL for very high and high risk, respectively). Concomitant high-dose statin therapy emerged as the primary predictor of LDL-C target attainment. Heterozygous familial hypercholesterolemia (HeFH), statin intolerance, and female gender were associated with a significant lower probability of achieving LDL-C target levels. Conclusions: Our analysis confirms that PCSK9mAb treatment is safe and effective, enabling 60% of our cohort to fully achieve the LDL-C guideline recommendations. The use of high-intensity statins emerged as a significant predictor of efficacy. Conversely, familial hypercholesterolemia and female gender were identified as predictors of therapeutic failure. Hence, it is crucial to address disparities in cardiovascular disease prevention between genders and to enhance strategies for managing elevated LDL-C in HeFH patients.
Probiotics in Traumatic Brain Injury: New Insights into Mechanisms and Future Perspectives
Authors: Pagkou Diamantoula , Kogias Evangelos , Foroglou Nikolaos , Kotzampassi Katerina
Published: August 2024

Traumatic brain injury (TBI) is a serious global public health issue, recognized as a chronic and progressive disease that can affect multiple organs, including the gastrointestinal (GI) tract. Research shows that there is a specific link between the GI tract and the central nervous system, termed the gut–brain axis, which consists of bidirectional exchange between these two. Several preclinical and clinical studies have demonstrated intestinal barrier dysfunction, intestinal inflammation and gut dysbiosis in patients with TBI. It is proven that probiotics can modulate the inflammatory process and modify gut microbiota. Numerous animal studies and human clinical trials have proven the effectiveness of selected bacterial strains as an adjuvant treatment in reducing inflammation, infection rates and time spent in intensive care of hospitalized patients suffering from brain injury. Thus, this review summarizes the current evidence regarding the beneficial effects of probiotic administration in patients suffering from TBI-related complications. This review will help identify novel therapeutic strategies in the future as probiotics have an extensive history of apparently safe use.
Chronic Right Ventricular Pacing Post-Transcatheter Aortic Valve Replacement Attenuates the Benefit on Left Ventricular Function
Authors: Chao Chieh-Ju , Mandale Deepa , Farina Juan M. , Abdou Merna , Rattanawong Pattara , Girardo Marlene , Agasthi Pradyumma , Ayoub Chadi , Alkhouli Mohammad , Eleid Mackram , Fortuin F. David , Sweeney John P. , Pollak Peter , Sabbagh Abdallah El , Holmes David R. , Arsanjani Reza , Naqvi Tasneem Z.
Published: August 2024

Background: Conduction abnormality post-transcatheter aortic valve implantation (TAVI) remains clinically significant and usually requires chronic pacing. The effect of right ventricular (RV) pacing post-TAVI on clinical outcomes warrants further studies. Methods: We identified 147 consecutive patients who required chronic RV pacing after a successful TAVI procedure and propensity-matched these patients according to the Society of Thoracic Surgeons (STS) risk score to a control group of patients that did not require RV pacing post-TAVI. We evaluated routine echocardiographic measurements and performed offline speckle-tracking strain analysis for the purpose of this study on transthoracic echocardiographic (TTE) images performed at 9 to 18 months post-TAVI. Results: The final study population comprised 294 patients (pacing group n = 147 and non-pacing group n = 147), with a mean age of 81 ± 7 years, 59% male; median follow-up was 354 days. There were more baseline conduction abnormalities in the pacing group compared to the non-pacing group (56.5% vs. 41.5%. p = 0.01). Eighty-eight patients (61.6%) in the pacing group required RV pacing due to atrioventricular (AV) conduction block post-TAVI. The mean RV pacing burden was 44% in the pacing group. Left ventricular ejection fraction (LVEF) was similar at follow-up in the pacing vs. non-pacing groups (57 ± 13.0%, 59 ± 11% p = 0.31); however, LV global longitudinal strain (−12.7 ± 3.5% vs. −18.8 ± 2.7%, p < 0.0001), LV apical strain (−12.9 ± 5.5% vs. 23.2 ± 9.2%, p < 0.0001), and mid-LV strain (−12.7 ± 4.6% vs. −18.7 ± 3.4%, p < 0.0001) were significantly worse in the pacing vs. non-pacing groups. Conclusions: Chronic RV pacing after the TAVI procedure is associated with subclinical LV systolic dysfunction within 1.5 years of follow-up.
Effects of SGLT2 Inhibitors on Cardiac Mechanics in Hispanic and Black Diabetic Patients
Authors: Moras Errol , Shrivastav Rishi , Gandhi Kruti D. , Bandyopadhyay Dhrubajyoti , Isath Ameesh , Goel Akshay , Bella Jonathan N. , Contreras Johanna
Published: August 2024

Background: Clinical trials demonstrating improved cardiovascular outcomes with SGLT2 inhibitors have often had limited representation from Black and Hispanic populations. While the mechanisms of action are not well known, ethnicity- or gender-based receptor physiology may render SGLT2 inhibitors a better agent in certain populations over others. Methods: A medical records query yielded diabetic patients initiated on SGLT2 inhibitors between 2013 and 2020. Patients with coronary artery disease, cardiac arrhythmias, and heart failure were excluded. Transthoracic echocardiographic studies (TTE) before and after starting SGLT2 inhibitors were analyzed, and post-processing left ventricular global longitudinal strain (LV GLS) analysis was also performed on each echocardiographic study. Univariate outliers and patients with missing data were excluded. Results: Among 94 patients with TTE (mean age 60.7 years; 68% Hispanics, 22.3% Blacks; median follow up of 7 months), there were significant improvements in the mean LV GLS (−15.3 vs. −16.5; p = 0.01), LV mass (LVM) (198.4 ± 59.6 g vs. 187.05 ± 50.6 g; p = 0.04), and LV mass index (LVMI) (100.6 ± 26.6 g/m2 vs. 94.3 ± 25.4 g/m2; p = 0.03) before and after initiating SGLT2 inhibitors but no significant change in the ratio (MV E/E’) of peak early diastolic mitral flow velocity (E) and spectral pulsed-wave Doppler-derived early diastolic velocity from the septal mitral annulus (E’) (12.5 ± 5.7 vs. 12.7 ± 4.8; p = 0.38). Changes in HbA1c (r2 = 0.82; p = 0.026), LVM (r2 = 0.20; p = 0.04), and LVMI (r2 = 0.20; p = 0.04) were found to be independently associated with changes in values of LV GLS on follow-up echocardiograms, when compared to the pre-medication LV GLS number. Conclusion: Non-White diabetic patients receiving SGLT2 inhibitors against a backdrop of other cardioprotective medications demonstrate significant improvements in LV remodeling and LV GLS, driven in part by an improvement in glycemic control. Large, prospective studies are needed to explore the differences in the therapeutic actions of SGLT2 inhibitors among different populations.