Short-term effectiveness and side effects of ketogenic diet for drug-resistant epilepsy in children with genetic epilepsy syndromes.

Background: Drug-resistant epilepsy (DRE) impacts a significant portion, one-third, of individuals diagnosed with epilepsy. In such cases, exploring non-pharmacological interventions are crucial, with the ketogenic diet (KD) standing out as a valuable option. KD, a high-fat and low-carb dietary approach with roots dating back to the 1920s for managing DRE, triggers the formation of ketone bodies and modifies biochemistry to aid in seizure control. Recent studies have increasingly supported the efficacy of KD in addressing DRE, showcasing positive outcomes. Furthermore, while more research is needed, limited data suggests that KD May also be beneficial for specific genetic epilepsy syndromes (GESs). Objective: This study aimed to assess the short-term efficacy of KD among pediatric patients diagnosed with GESs. Materials and methods: This is a multi-center retrospective analysis of pediatric patients with GESs diagnosed using next-generation sequencing. The enrolled patients followed the keto-clinic protocol, and the KD efficacy was evaluated at 3, 6, and 12-month intervals based on seizure control and compliance. The collection instrument included demographic, baseline, and prognostic data. The collected data was coded and analyzed promptly. Results: We enrolled a cohort of 77 patients with a mean current age of 7.94 ± 3.83 years. The mean age of seizure onset was 15.5 months. Notably, patients experienced seizures at a younger age tended to have less positive response to diet. Overall, 55 patients responded favorably to the diet (71.4%) while 22 patients (28.6%) showed no improvement. Patients with genetic etiology showed a significantly more favorable responses to the dietary intervention. Patients with Lennox-Gastaut syndrome showed the most significant improvement (14/15) followed by patients with Dravet syndrome (6/8), and West syndrome (3/4). The number of used anti-seizure medications also played a significant role in determining their response to the diet. While some patients experienced mild adverse events, the most common being constipation, these occurrences were not serious enough to necessitate discontinuation of the diet. Conclusion: The study revealed a high improvement rate in seizure control, especially among younger patients and those with later seizure onset. The success of dietary treatment hinges greatly on early intervention and the patient's age. Certain genetic mutations responded favorably to the KD, while efficacy varied among various genetic profiles.

Corrigendum: Antibiotic resistance: One Health One World outlook.

[This corrects the article DOI: 10.3389/fcimb.2021.771510.].

Utilization of opium poppy seed oil for biodiesel production: A parametric characterization and statistical optimization.

Consuming traditional petroleum-derived diesel fuel has long been associated with issues such as the depletion of natural energy resources. To solve these challenges, an alternate source like as biodiesel is an appealing option. Seed oils have long been recognized as an abundant and diverse source of biodiesel. In this study, poppy seed oil from the poppy (Papaver somniferum) was investigated for biodiesel production. Poppy seed biodiesel was generated and refined using acid-pretreated esterification with sulphuric acid prior to transesterification, as well as single-step alkaline catalyzed transesterification with methanol and potassium hydroxide. Finally, the percentage yield was compared. Using Statistica, the Box-Behnken design was applied to optimize process variables like time, temperature, catalyst concentration, and methanol-oil ratio to produce maximum yield. The relationship of process variables was also shown with the help of the Response Surface Methodology. A maximum yield of 94.87 % was obtained at optimized conditions, i.e., 90min reaction time, 60 °C of temperature, 0.25 mg of catalyst concentration, and 3v/v% alcohol-oil ratio. The fuel properties of biodiesel produced, such as acid value, moisture content, saponification value, iodine value, specific gravity, percentage of free fatty acids, refractive index, viscosity, boiling point, and peroxide value, were measured and compared with the American Society for Testing and Materials (ASTM) D6751 and European Standards (EN) 14214. Further results were studied and discussed using Fourier Transfer Infrared (FTIR) analysis, which showed maximum similarity of raw material to formed biodiesel. Gas Chromatography-Mass Spectrometry (GC-MS) analysis was performed to identify and quantify various fatty acid methyl esters. The results obtained were in accordance with various international standards for biodiesel fuel. Thus, poppy seeds can be used to obtain biodiesel.

Predictive modeling for the adsorptive and photocatalytic removal of phenolic contaminants from water using artificial neural networks.

Numerous harmful phenolic contaminants are discharged into water that pose a serious threat to environment where two of the most important purification methodologies for the mitigation of phenolic contaminants are adsorption and photocatalysis. Besides cost, each process has drawbacks in terms of productivity, environmental impact, sludge creation, and the development of harmful by-products. To overcome these limitations, the modeling and optimization of water treatment methods is required. Artificial Intelligence (AI) is employed for the interpretation of treatment-based processes due to powerful learning, simplicity, high estimation accuracy, effectiveness, and improvement of process efficiency where artificial neural networks (ANNs) are most frequently employed for predicting and analyzing the efficiency of processes applied for the mitigation of these phenolic contaminants from water. ANNs are superior to conventional linear regression models because the latter are incapable of dealing with non-linear systems. ANNs can also reduce the operational cost of treating phenol-contaminated water. A correlation coefficient of >0.99 can be achieved using ANN with enhanced phenol mitigation percentage accuracy generally ranging from 80 % to 99.99 %. Using ANN optimization, the maximum phenol mitigation efficiencies achieved were 99.99 % for phenol, 99.93 % for bisphenol A, 99.6 % for nonylphenol, 97.1 % for 2-nitrophenol, 96.6 % for 4-chlorophenol and 90 % for 2,6-dichlorophenol. In numerous ANN models, Levenberg-Marquardt backpropagation algorithm for training was employed using MATLAB software. This study overviews their employment and application for optimization and modeling of removal processes and explicitly discusses the important input and output parameters necessary for better performance of the system. The comparison of ANNs with other AI techniques revealed that ANNs have better predictability for mitigation of most of the phenolic contaminants. Furthermore, several challenges and future prospects have also been discussed.

Mirabegron Versus Placebo and Other Therapeutic Modalities in the Treatment of Patients with Overactive Bladder Syndrome-A Systematic Review.

BACKGROUND AND OBJECTIVE: The prevalence of overactive bladder (OAB) increases with age. Mirabegron and other drugs are used for the management of patients with OAB. To evaluate mirabegron versus other treatments for overactive bladder syndrome (OAB). METHODS: This randomised controlled trial (RCT)-based systematic review (CRD42020200394) was conducted following the 2020 Preferred Reporting Items for Systematic Reviews and Meta-analyses statement, with standards reported in the Cochrane Handbook for Systematic Reviews of Interventions. KEY FINDINGS AND LIMITATIONS: We included 28 RCTs (n = 27 481 adults), comparing the following: mirabegron 25 mg versus placebo (n = 8798; six RCTs): significant changes in urgency urinary incontinence (mean difference [MD] -0.41, 95% confidence interval [CI] -0.56 to -0.26), total incontinence (MD -0.47, 95% CI -0.63 to -0.30), and nocturia (MD -0.10, 95% CI -0.17 to -0.02), and mirabegron 50 mg versus placebo (n = 14 933; 12 RCTs): significant changes in urgency urinary incontinence (MD -0.41, 95% CI -0.52 to -0.31), urgency (MD -0.49, 95% CI -0.64 to -0.33), total incontinence (MD -0.44, 95% CI -0.55 to -0.33), favouring mirabegron 25/50 mg; mirabegron 50 mg versus tolterodine 4 mg (n = 8008; five RCTs): significant changes in micturition (MD -0.16, 95% CI -0.31 to -0.02) and overall adverse events (AEs; odds ratio [OR] 0.71, 95% CI 0.59-0.86), favouring mirabegron 50 mg; mirabegron 50 mg versus solifenacin 5 mg (n = 8911; four RCTs): significant changes in voided volume/micturition in millilitres (MD -7.77, 95% CI -12.93 to -2.61), favouring mirabegron 50 mg; and mirabegron 50 mg versus oxybutynin 73.5 mg (n = 302; one RCT): significant changes in overall AEs (OR 0.02, 95% CI 0.00-0.16), favouring mirabegron 50 mg. CONCLUSIONS AND CLINICAL IMPLICATIONS: Mirabegron is effective, safe, and well tolerated. Coadministration with anticholinergics provides an advantageous additive effect without a higher occurrence of side effects. PATIENT SUMMARY: Mirabegron is effective, safe, and well tolerated for treating overactive bladder. When used in conjunction with anticholinergic medications, it provides extra benefits without causing more side effects.

Co-exposure to microplastics and soil pollutants significantly exacerbates toxicity to crops: Insights from a global meta and machine-learning analysis.

Environmental contamination of microplastics (MPs) is ubiquitous worldwide, and co-contamination of arable soils with MPs and other pollutants is of increasing concern, and may lead to unexpected consequences on crop production. However, the overall implications of this combined effect, whether beneficial or detrimental, remain a subject of current debate. Here, we conducted a global meta and machine-learning analysis to evaluate the effects of co-exposure to MPs and other pollutants on crops, utilizing 3346 biological endpoints derived from 68 different studies. Overall, compared with control groups that only exposure to conventional soil contaminants, co-exposure significantly exacerbated toxicity to crops, particularly with MPs intensifying adverse effects on crop morphology, oxidative damage, and photosynthetic efficiency. Interestingly, our analysis demonstrated a significant reduction in the accumulation of pollutants in the crop due to the presence of MPs. In addition, the results revealed that potential adverse effects were primarily associated with crop species, MPs mass concentration, and exposure duration. Our study reaffirms the substantial repercussions of MPs as emerging pollutants on crops within the context of integrated pollution, providing novel insights into improving sustainability in agro-ecosystems.

Genome-Wide Mapping of Consanguineous Families Confirms Previously Implicated Gene Loci and Suggests New Loci in Specific Language Impairment (SLI).

Specific language impairment (SLI) is a developmental disorder with substantial genetic contributions. A genome-wide linkage analysis and homozygosity mapping were performed in five consanguineous families from Pakistan. The highest LOD scores of 2.49 at 12p11.22-q11.21 in family PKSLI-31 and 1.92 at 6p in family PKSLI-20 were observed. Homozygosity mapping showed a loss of heterozygosity on 1q25.3-q32.2 and 2q36.3-q37.3 in PKSLI-20. A loss of heterozygosity mapped, in PKSLI-31 and PKSLI-34 flanks, NFXL1 and CNTNAP2, which are genes previously identified in SLI. Our findings report novel SLI loci and corroborate previously reported SLI loci, indicating the utility of a family-based approach.

Antioxidant Activity of Protein Hydrolysates from Redlip Mullet (Chelon haematocheilus) Muscle and Byproducts.

Fish muscle and byproducts represent a valuable source of bioactive compounds, with their protein hydrolysates exhibiting noteworthy antioxidant properties. This study assessed the antioxidant activity of protein hydrolysates derived from the muscle and byproducts of redlip mullet (Chelon haematocheilus), utilizing different proteases (Neutrase, Alcalase, and Protamex). Hydrolysates were prepared from various parts of the fish, including muscle (white and red meat) and byproducts (frames, head, viscera, fins, skin, and scales). The enzymatic hydrolysis resulted in the highest degree of hydrolysis, achieving 83.24 ± 1.45% for skin at 60 min and 82.14 ± 4.35% for head at 30 min, when treated with Neutrase. Frames treated with Neutrase exhibited the highest protein concentration, measured at 1873.01 ± 71.11 µg/mL at 15 min. Significantly, skin hydrolysates treated with Protamex showed the highest DPPH• scavenging activity (70.07 ± 3.99% at 120 min), while those treated with Alcalase demonstrated the highest ABTS• scavenging activity (93.47 ± 0.02% at 15 min). The highest superoxide dismutase (SOD) activity (92.01 ± 1.47%) was observed in head hydrolysates treated with Protamex after 90 min. These results suggest that C. haematocheilus protein hydrolysates possess significant antioxidant activity within a short time frame of less than 120 min.

Prevalence of Iron Deficiency Among Patients With Helicobacter pylori Infection at a Tertiary Care Hospital.

Background and objective Iron deficiency and iron deficiency anemia are related but distinct conditions. Iron deficiency refers to a state where the body has insufficient iron stores, which can lead to anemia if not addressed. Iron deficiency anemia, on the other hand, is a more severe condition where the lack of iron has resulted in decreased hemoglobin levels, impacting oxygen transport in the blood. Helicobacter pylori (H. pylori) infection can contribute to iron deficiency through mechanisms such as chronic gastrointestinal bleeding and impaired iron absorption, potentially progressing to iron deficiency anemia. The objective of this study was to determine the prevalence of iron deficiency, including its potential progression to iron deficiency anemia, among patients diagnosed with H. pylori infection. Methodology This cross-sectional study was conducted at Bolan Medical Complex Hospital, Quetta, and included 200 patients diagnosed with H. pylori infection via endoscopic biopsy or urea breath test, from January to June 2023. Participants were aged 18 years and older, excluding those with chronic diseases affecting iron metabolism, current iron supplement users, and pregnant women. Data were collected through questionnaires and medical records, and blood samples were analyzed for serum ferritin and hemoglobin levels. Statistical analysis included chi-square tests and logistic regression was performed in SPSS (version 27; IBM Corp, Armonk, NY); p-value <0.05 was significant. Results Out of 200 patients, 80 (40%) were iron deficient. The prevalence was highest among those over 60 years (n = 15, 75%) compared to the 18-30 age group (n = 10, 20%). Males had a slightly higher prevalence of iron deficiency (n = 50, 45.5%) compared to females (n = 30, 33.3%). Patients with H. pylori infection for more than three years exhibited a higher prevalence of iron deficiency (n = 30, 50%) compared to those with less than one year of infection (10/60, 16.7%). Dietary habits also played a role, with vegetarians showing a higher prevalence (n = 20, 50%) compared to non-vegetarians (n = 60, 37.5%). Hemoglobin levels were significantly lower in iron-deficient participants, averaging 10.8 ± 0.9 g/dL, and logistic regression indicated significant associations between iron deficiency and both age (OR = 1.05, p = 0.001) and infection duration (OR = 1.10, p < 0.001). Conclusions The study revealed a significant prevalence of iron deficiency among H. pylori-infected patients, particularly in older adults, males, those with longer infection duration, and vegetarians. The findings underscore the need for routine monitoring and targeted treatment of iron deficiency in this population.

Transplanting network pharmacology technology into food science research: A comprehensive review on uncovering food-sourced functional factors and their health benefits.

Network pharmacology is an emerging interdisciplinary research method. The application of network pharmacology to reveal the nutritional effects and mechanisms of active ingredients in food is of great significance in promoting the development of functional food, facilitating personalized nutrition, and exploring the mechanisms of food health effects. This article systematically reviews the application of network pharmacology in the field of food science using a literature review method. The application progress of network pharmacology in food science is discussed, and the mechanisms of functional factors in food on the basis of network pharmacology are explored. Additionally, the limitations and challenges of network pharmacology are discussed, and future directions and application prospects are proposed. Network pharmacology serves as an important tool to reveal the mechanisms of action and health benefits of functional factors in food. It helps to conduct in-depth research on the biological activities of individual ingredients, composite foods, and compounds in food, and assessment of the potential health effects of food components. Moreover, it can help to control and enhance their functionality through relevant information during the production and processing of samples to guarantee food safety. The application of network pharmacology in exploring the mechanisms of functional factors in food is further analyzed and summarized. Combining machine learning, artificial intelligence, clinical experiments, and in vitro validation, the achievement transformation of functional factor in food driven by network pharmacology is of great significance for the future development of network pharmacology research.

CircRNA-Phf21a_0002 promotes pyroptosis to aggravate hepatic ischemia/ reperfusion injury by sponging let-7b-5p.

Hepatic ischemia‒reperfusion injury is a common injury in liver surgery and liver transplantation that can lead to liver function damage, including oxidative stress, apoptosis, autophagy and inflammatory reactions. Pyroptosis is a type of inflammatory programmed cell death that has been implicated in ischemia‒reperfusion injury-associated inflammatory reactions. Although circular RNAs can regulate cell death in hepatic ischemia‒reperfusion injury, their relationship with pyroptosis remains unclear. Therefore, this study aimed to investigate the effect of circular RNA on pyroptosis in hepatic ischemia‒reperfusion injury. We constructed a mouse hepatic ischemia‒reperfusion injury model for circular RNA sequencing and obtained 40 circular RNAs with significant differential expression, of which 39 were upregulated and 1 was downregulated. Subsequently, the endogenous competitive RNA network was constructed using TarBase, miRTarBase, TargetScan, RNAhybrid, and miRanda. Gene Set Enrichment Analysis, Kyoto Encyclopedia of Genes and Genomes and Gene Ontology functional analyses of downstream target genes revealed that circRNA-Phf21a_0002 might affect pyroptosis by regulating the mTOR signaling pathway and Bach1 by sponging let-7b-5p. The overexpression plasmid upregulated the expression of circRNA-Phf21a_0002 in a hypoxia/reoxygenation model, which aggravated pyroptosis in AML12 cells and apoptosis and necrosis of hepatocytes. Next, we investigated the underlying mechanism and found that circRNA-Phf21a_0002 enabled the expression of Bach1 through sponging of let-7b-5p. The aggravation of pyroptosis via overexpression of circRNA-Phf21a_0002 was reversed by let-7b-5p mimics in hypoxia/reoxygenation-subjected AML12 cells. Collectively, our study clarifies that circRNA-Phf21a_0002 aggravates the pyroptosis of hepatocytes related to ischemia-reperfusion by sponging let-7b-5p. These findings provide new molecular mechanisms and novel biomarkers for follow-up treatment.

Investigation of therapeutic potential of the Il24-p20 fusion protein against breast cancer: an in-silico approach.

Targeted delivery of therapeutic anticancer chimeric molecules enhances drug efficacy. Numerous studies have focused on developing novel treatments by employing cytokines, particularly interleukins, to inhibit the growth of cancer cells. In the present study, we fused interleukin 24 with the tumor-targeting peptide P20 through a rigid linker to selectively target cancer cells. The secondary structure, tertiary structure, and physicochemical characteristics of the constructed chimeric IL-24-P20 protein were predicted by using bioinformatics tools. In-silico analysis revealed that the fusion construct has a basic nature with 175 amino acids and a molecular weight of 20 kDa. By using the Rampage and ERRAT2 servers, the validity and quality of the fusion protein were evaluated. The results indicated that 93% of the chimeric proteins contained 90.1% of the residues in the favoured region, resulting in a reliable structure. Finally, docking and simulation studies were conducted via ClusPro and Desmond Schrödinger, respectively. Our results indicate that the constructed fusion protein exhibits excellent quality, interaction capabilities, validity, and stability. These findings suggest that the fusion protein is a promising candidate for targeted cancer therapy.

Innovative player evaluation: Dual-possibility Pythagorean fuzzy hypersoft sets for accurate international football rankings.

This study introduces an advanced approach for ranking international football players, addressing the inherent uncertainties in performance evaluations. By integrating dual possibility theory and Pythagorean fuzzy sets, the model accommodates varying degrees of ambiguity and imprecision in player attributes. Additionally, the use of hypersoft set theory enriches the analysis by capturing the multifaceted nature of player evaluations. The proposed aggregation operators refine the synthesis of diverse information sources, leading to a comprehensive and nuanced assessment. This research significantly enhances player evaluation methodologies, providing a more adaptable framework for a fair assessment of international football talent. A practical example illustrates the application of dual-possibility Pythagorean fuzzy hypersoft sets (DP-PFHSS). A numerical technique is proposed for solving multi-criteria decision-making (MCDM) challenges with known dual possibility information using the proposed aggregation operators. This decision-making algorithm effectively determines a football player's worth, contributing to the overall ranking and evaluation process. The approach aids in scouting and recruitment by facilitating talent identification and informed player signings. Graphical analysis, comparing existing and proposed methods using average and geometric operators, demonstrates the superiority of the proposed approach in the players evaluation, indicating that F 1 is in the top ranking.

Enhancement in the antibacterial activity of Rifaximin by delivery through gelatin nanoparticles.

OBJECTIVES: Bacterial infections are a noteworthy global health concern that necessitates the development of new strategies to enhance the potency and efficacy of antibiotics. Rifaximin (RFX), a broad-spectrum antibiotic, exhibits promising antibacterial activity against several bacterial strains. However, its insolubility and impermeability impede the exploitation of its full potential. The objective of the current study is to overcome the inherent caveats of RFX to exploit its maximum potential. SIGNIFICANCE: The exploitation of the full potential of antibiotics is necessary for reduction in their dosage and to minimize antibiotic pollution. This is a preliminary study aiming for maximum utilization of RFX at the target site and reduction in its release in unmetabolized form. METHODS: Gelatin is a biopolymer that has gained significant attention for biomedical applications owing to its inherent biocompatibility and biodegradability. In this study, bovine gelatin nanoparticles (BGNPs) were fabricated by the self-assembly method for their application as a carrier of RFX to enhance its antibacterial activity. The study employs a comprehensive range of experimental techniques to characterize the fabricated BGNPs such as DLS, Zeta Potential, FT-IR, AFM, SEM-EDX, and UV-Vis spectrophotometry. RESULTS: The average size of the fabricated BGNPs was 100 nm with a zeta potential value of -15.3 mV. The loading of RFX on BGNPs rendered an increase in its size to 136 nm with a zeta potential value of -16 mV. In-vitro assays and microscopic analyses were conducted to compare the antibacterial efficacy of RFX and RFX@BGNPs. An excellent loading capacity followed by sustained release of RFX from RFX@BGNPs rendered a significant enhancement in its pharmaceutical efficacy. The release of RFX from RFX@BGNPs followed the Higuchi and Korsmeyer-Peppas models. The antibacterial efficacy of RFX against Staphylococcus aureus has doubled by delivery through RFX@BGNPs, assessed by inhibitory and biofilm inhibitory assays. The enhancement in the antibacterial efficiency was further endorsed by SEM and microscopic imaging of the control and treated bacterial colonies. CONCLUSION: The study demonstrates an enhancement in the antimicrobial efficacy of RFX by its delivery in the form of RFX@BGNPs to exploit its full potential for practical applications.

Social dimensions of climate-induced flooding in Jakarta (Indonesia): The role of non-point source pollution.

Because of its low-lying location, urbanization, and inadequate infrastructure, Jakarta (Indonesia) has experienced an increase in annual flooding events, rising from an average of five significant floods per year in the 1990s to over 20 annually (2010-2020). With climate change exacerbating extreme weather events, Jakarta encounters escalating risks of flooding. Although the recurrent flooding is exacerbated by non-point source (NPS) of pollution such as urban runoff and agricultural discharge that contribute to 40% of total pollutants leading to flood-related issues in Jakarta, none has investigated this research gap. To reflect its novelty, this work explores the implications of climate change on the annual flooding in Jakarta by focusing on NPS and analyzes their impacts from social perspectives. This work also underscores the implications of flooding on livelihoods, health, and social cohesion in Jakarta. Focus group discussion with affected residents was used to shed light on the coping strategies employed in response to recurrent floods, ranging from community-based initiatives to reliance on informal networks. The empirical findings show that the implications of flooding extend beyond physical damages. Displacement of communities, loss of livelihoods, disruption of essential services, and increased health risks are among the social impacts experienced by local residents. Vulnerable populations, including low-income communities residing in informal settlements, bear their consequences. Economic losses from flooding amount to USD 500 million annually, impacting over 1 million residents. However, recent interventions have led to a 15% reduction in peak flood levels and a 20% reduction in flood duration in affected areas. Community resilience has also improved, with a 25% increase in flood insurance coverage and a 20% rise in community response initiatives. Overall, this study highlights that climate change exacerbates annual flooding in Jakarta, significantly impacting vulnerable communities through NPS pollution. Addressing the challenges requires integrated approaches combining effective pollution control, resilient infrastructure, and community engagement to mitigate social and long-term environmental impacts. PRACTITIONER POINTS: Climate-induced flooding disproportionately affects vulnerable communities in Jakarta. Non-point source pollution from urban runoff contributes to the severity of flooding in Jakarta. Waterborne diseases, disruption of livelihoods, and reduced access to clean water are major concerns identified in the study. The study highlights the importance of community-based adaptation strategies to mitigate the impact of flooding and pollution.

De novo assembly of sialotranscriptome of Hyalomma anatolicum and insights into expression dynamics in response to Theileria annulata infection.

Hyalomma anatolicum is a tick of significant one-health importance due to its role as a vector for various pathogens affecting humans, animals and the environment, such as Theileria annulata, which causes tropical theileriosis in cattle, leading to severe economic losses. When infected with pathogens like T. annulata, the salivary glands of H. anatolicum undergo gene expression changes, secrete modified proteins and activate immune responses, all of which facilitate pathogen survival and transmission by modulating the host immune response and optimizing conditions for pathogen development. Understanding these responses is crucial for developing control strategies for tick-borne diseases. To understand the interaction between H. anatolicum and T. annulata, we performed a differential gene expression analysis of H. anatolicum salivary glands. An average of approximately 25 million raw sequencing reads were generated in each replicate using Illumina Sequencing. The sequenced reads were de novo assembled and the assembled transcriptome yielded approximately 50,231 non-redundant transcripts after clustering with CD-HIT using a sequence identity of 95% and alignment coverage of 90%. The assembly quality was evaluated with BUSCO analysis and found to be 86% complete using the Arachnida dataset and then blasted against non-redundant protein sequence database from NCBI followed by counting of reads and differential expression analysis. Overall, around 2400 and 400 genes were found differentially expressed with logFC > 1 and logFC > 2 respectively at FDR < 0.05. Top up-regulated genes included Calpain, Papilin, Neprilysin, and Ankyrin repeat-containing protein. Top down-regulated genes included Scoloptoxin, and Selenoprotein S and other uncharacterized proteins. Many other up-regulated proteins with high significance were uncharacterized suggesting room for further H. anatolicum functional and structural characterization studies. To our best knowledge, this is the first study of H. anatolicum sialotranscriptome which greatly contributes to sialotranscriptome information not only as sequence database but also indicates the potential targets for development of vaccine against ticks and transmission-blocking vaccines against T. annulata.

Single cell RNA-sequencing delineates CD8+ tissue resident memory T cells maintaining rejection in liver transplantation.

Rationale: Understanding the immune mechanisms associated with liver transplantation (LT), particularly the involvement of tissue-resident memory T cells (TRMs), represents a significant challenge. Methods: This study employs a multi-omics approach to analyse liver transplant samples from both human (n = 17) and mouse (n = 16), utilizing single-cell RNA sequencing, bulk RNA sequencing, and immunological techniques. Results: Our findings reveal a comprehensive T cell-centric landscape in LT across human and mouse species, involving 235,116 cells. Notably, we found a substantial increase in CD8+ TRMs within rejected grafts compared to stable ones. The elevated presence of CD8+ TRMs is characterised by a distinct expression profile, featuring upregulation of tissue-residency markers (CD69, CXCR6, CD49A and CD103+/-,), immune checkpoints (PD1, CTLA4, and TIGIT), cytotoxic markers (GZMB and IFNG) and proliferative markers (PCNA and TOP2A) during rejection. Furthermore, there is a high expression of transcription factors such as EOMES and RUNX3. Functional assays and analyses of cellular communication underscore the active role of CD8+ TRMs in interacting with other tissue-resident cells, particularly Kupffer cells, especially during rejection episodes. Conclusions: These insights into the distinctive activation and interaction patterns of CD8+ TRMs suggest their potential utility as biomarkers for graft rejection, paving the way for novel therapeutic strategies aimed at enhancing graft tolerance and improving overall transplant outcomes.

Imitation-inhibition training can reduce the observation-inflation effect in face-to-face scenarios.

Observing others performing an action can lead to false memories of self-performance-the observation-inflation effect. Previous research has indicated that this phenomenon might impact the memory of actions in real-world interactions. However, whether direct observation without interaction can lead to observation inflation remains unclear. In Experiment 1, participants passively observed the experimenter performing actions live. In subsequent memory tests, they indeed reported false memories regarding their performances. Building on this, Experiment 2 investigated the causes of the observation-inflation effect induced by "real" actions. Participants underwent imitation-inhibition training with the individuals they observed previously. The results revealed that participants who completed imitation-inhibition training reported fewer false memories in memory tests than those who completed imitation training. These findings suggest that even passive observation of "real" actions can lead to observation inflation, and the simulation of others' actions by individuals may be a potential underlying cause of their occurrence in real-life situations.

Humoral and cellular immunity in response to an in silico-designed multi-epitope recombinant protein of Theileria annulata.

Tropical theileriosis is a lymphoproliferative disease caused by Theileria annulata and is transmitted by Ixodid ticks of the genus Hyalomma. It causes significant losses in livestock, especially in exotic cattle. The existing methods for controlling it, chemotherapeutic agents and a vaccine based on an attenuated schizont stage parasite, have several limitations. A promising solution to control this disease is the use of molecular vaccines based on potential immunogenic proteins of T. annulata. For this purpose, we selected five antigenic sequences of T. annulata, i.e. SPAG-1, Tams, TaSP, spm2, and Ta9. These were subjected to epitope prediction for cytotoxic T lymphocytes, B-cells, and helper T lymphocytes. CTL and B-cell epitopes with a higher score whereas those of HTL with a lower score, were selected for the construct. A single protein was constructed using specific linkers and evaluated for high antigenicity and low allergenicity. The construct was acidic, hydrophobic, and thermostable in nature. Secondary and tertiary structures of this construct were drawn using the PSIPRED and RaptorX servers, respectively. A Ramachandran plot showed a high percentage of residues in this construct in favorable, allowed, and general regions. Molecular docking studies suggested that the complex was stable and our construct could potentially be a good candidate for immunization trials. Furthermore, we successfully cloned it into the pET-28a plasmid and transformed it into the BL21 strain. A restriction analysis was performed to confirm the transformation of our plasmid. After expression and purification, recombinant protein of 49 kDa was confirmed by western blotting. An ELISA detected increased specific antibody levels in the sera of the immunized animals compared with the control group, and flow cytometric analysis showed a stronger cell-mediated immune response. We believe our multi-epitope recombinant protein has the potential for the large-scale application for disease prevention globally in the bovine population. This study will act as a model for similar parasitic challenges.