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Intracellular infection by a pathogen induces significant rewiring of host cell signaling and biological processes. Understanding how an intracellular pathogen such as Toxoplasma gondii modulates host cell metabolism with single-cell resolution has been challenged by the variability of infection within cultures and difficulties in separating host and parasite metabolic processes. A new study from Gallego-Lopez and colleagues (G. M. Gallego-López, E. C. Guzman, D. E. Desa, L. J. Knoll, M. C. Skala, mBio e00727-24, 2024, https://doi.org/10.1128/mbio.00727-24) applies a quantitative imaging approach to evaluate the host cell metabolism during intracellular infection with Toxoplasma . This study provides important insights into host metabolic responses to Toxoplasma infection and offers a valuable tool to dissect the mechanisms underlying parasite infection and pathophysiology., Competing Interests: The authors declare no conflict of interest.

The production of energy in the form of ATP by the mitochondrial ATP synthase must be tightly controlled. One well-conserved form of regulation is mediated via ATPase inhibitory factor 1 (IF1), which governs ATP synthase activity and gene expression patterns through a cytoprotective process known as mitohormesis. In apicomplexans, the processes regulating ATP synthase activity are not fully elucidated. Using the model apicomplexan Toxoplasma gondii , we found that knockout and overexpression of TgIF1, the structural homolog of IF1, significantly affected gene expression. Additionally, TgIF1 overexpression resulted in the formation of a stable TgIF1 oligomer that increased the presence of higher order ATP synthase oligomers. We also show that parasites lacking TgIF1 exhibit reduced mitochondrial cristae density, and that while TgIF1 levels do not affect growth in conventional culture conditions, they are crucial for parasite survival under hypoxia. Interestingly, TgIF1 overexpression enhances recovery from oxidative stress, suggesting a mitohormetic function. In summary, while TgIF1 does not appear to play a role in metabolic regulation under conventional growth conditions, our work highlights its importance for adapting to stressors faced by T. gondii and other apicomplexans throughout their intricate life cycles.

Apicomplexans, such as Plasmodium and Toxoplasma are obligate intracellular parasites that invade, replicate and finally EXIT their host cell. During replication within a parasitophorous vacuole (PV), the parasites establish an extensive F-actin-containing network that connects individual parasites and is required for material exchange, recycling and the final steps of daughter cell assembly. After multiple rounds of replication, the parasites exit the host cell involving multiple signalling cascades, disassembly of the network, secretion of microneme proteins and activation of the acto-myosin motor. Blocking the host cell EXIT process leads to the formation of large PVs, making the screening for genes involved in exiting the cell relatively straightforward. Given that apicomplexans are highly diverse from other eukaryotes, approximately 30% of all genes are annotated as hypothetical, some apicomplexan-specific factors are likely to be critical during EXIT. This motivated several labs to design and perform forward genetic and phenotypic screens using various approaches, such as random insertion mutagenesis, temperature-sensitive mutants and, more recently, CRISPR/Cas9-mediated targeted editing and conditional mutagenesis. Here we will provide an overview of the technological developments over recent years and the most successful stories that led to the identification of new critical factors in Toxoplasma gondii., (© 2023 The Authors. Molecular Microbiology published by John Wiley & Sons Ltd.)

Background: Toxoplasma gondii (T. gondii) is a widespread, zoonotic protozoan intracellular parasite with a complex life cycle, which can cause toxoplasmosis, a potentially serious disease. During the invasion process, T. gondii proteins first bind to the relevant host cell receptors, such as glycosaminoglycan molecule (GAG-binding motif), which is one of the main receptors for parasites or virus to infect host cells. However, research on TGME49_216510 (T. gondii Trx21), a protein from Toxoplasma gondii, is limited. Methods: Bioinformatics analysis of the Trx21 protein was performed firstly. And specific primers were then designed using the conserved domain and GAG-binding motif to amplify, express, and purify a fragment of the Trx21 protein. The purified Trx21-GST protein was used for antioxidant and cell adhesion experiments. Simultaneously, mice were immunized with Trx21-His to generate specific polyclonal antibodies for subcellular localization analysis. Results: The Trx21 protein, consisting of 774 amino acids, included a transmembrane region, three GAG-binding motifs, and a Thioredoxin-like domain. The recombinant Trx21-His protein had a molecular mass of about 31 kDa, while the Trx21-GST protein had a molecular mass of about 55 kDa, which was analyzed by SDS-PAGE and Western blot. Subcellular localization analysis by IFA revealed that Trx21 is predominantly distributed in the cytoplasm of T. gondii. Furthermore, Trx21 exhibited a protective effect on supercoiled DNA against metal-catalyzed oxidation (MCO) and demonstrated adhesion abilities to Vero cells. Conclusions: These results indicate that Trx21 plays an important role in host cell interaction and oxidative damage. [ABSTRACT FROM AUTHOR]

Rhoptries are specialized secretory organelles conserved across the Apicomplexa phylum, essential for host cell invasion and critical for subverting of host cellular and immune functions. They contain proteins and membranous materials injected directly into the host cells, participating in parasitophorous vacuole formation. Toxoplasma gondii tachyzoites harbor 8 to 12 rhoptries, 2 of which are docked to an apical vesicle (AV), a central element associated with a rhoptry secretory apparatus prior to injection into the host cell. This parasite is also equipped with 5 to 6 microtubule-associated vesicles, presumably serving as AV replenishment for iterative rhoptry discharge. Here, we characterized a rhoptry protein, rhoptry discharge factor 3 (RDF3), crucial for rhoptry discharge and invasion. RDF3 enters the secretory pathway, localizing near the AV and associated with the rhoptry bulb. Upon invasion, RDF3 dynamically delocalizes, suggesting a critical role at the time of rhoptry discharge. Cryo-electron tomography analysis of RDF3-depleted parasites reveals irregularity in microtubule-associated vesicles morphology, presumably impacting on their preparedness to function as an AV. Our findings suggest that RDF3 is priming the microtubule-associated vesicles for rhoptry discharge by a mechanism distinct from the rhoptry secretory apparatus contribution. Regulated secretion of rhoptry factors is essential for host cell invasion by Apicomplexan parasites. This study identifies the protein rhoptry discharge factor 3 (RDF3) of Toxoplasma gondii as crucial for invasion, by priming microtubule-associated vesicles for rhoptry discharge. [ABSTRACT FROM AUTHOR]

The apicomplexan intracellular parasite Toxoplasma gondii is a major food borne pathogen that is highly prevalent in the global population. The majority of the T. gondii proteome remains uncharacterized and the organization of proteins into complexes is unclear. To overcome this knowledge gap, we used a biochemical fractionation strategy to predict interactions by correlation profiling. To overcome the deficit of high-quality training data in non-model organisms, we complemented a supervised machine learning strategy, with an unsupervised approach, based on similarity network fusion. The resulting combined high confidence network, ToxoNet, comprises 2,063 interactions connecting 652 proteins. Clustering identifies 93 protein complexes. We identified clusters enriched in mitochondrial machinery that include previously uncharacterized proteins that likely represent novel adaptations to oxidative phosphorylation. Furthermore, complexes enriched in proteins localized to secretory organelles and the inner membrane complex, predict additional novel components representing novel targets for detailed functional characterization. We present ToxoNet as a publicly available resource with the expectation that it will help drive future hypotheses within the research community. Author summary: Toxoplasma gondii is a food-borne parasite that chronically infects 1 in 3 worldwide. It can cause blindness in children and is life-threatening during pregnancy and to those with compromised immune systems. During infection, Toxoplasma relies on the coordination of suites of proteins that allow it to invade host cells and persist. In this study we combined a sophisticated biochemical separation platform with a novel machine learning strategy to build a global roadmap that describes how parasite proteins work together as molecular machines to help drive infection. Our roadmap, which we term ToxoNet, details 2,063 connections between 652 proteins. These include proteins representing parasite-specific innovations that help Toxoplasma perform essential functions such as the generation of energy. Further, in addition to capturing proteins already implicated in host cell invasion pathways, we identified many additional components of these pathways that represent novel targets for future drug development strategies. [ABSTRACT FROM AUTHOR]

The cell surface of Toxoplasma gondii is rich in glycoconjugates which hold diverse and vital functions in the lytic cycle of this obligate intracellular parasite. Additionally, the cyst wall of bradyzoites, that shields the persistent form responsible for chronic infection from the immune system, is heavily glycosylated. Formation of glycoconjugates relies on activated sugar nucleotides, such as uridine diphosphate N-acetylglucosamine (UDP-GlcNAc). The glucosamine-phosphate-N-acetyltransferase (GNA1) generates N-acetylglucosamine-6-phosphate critical to produce UDP-GlcNAc. Here, we demonstrate that downregulation of T. gondii GNA1 results in a severe reduction of UDP-GlcNAc and a concomitant drop in glycosylphosphatidylinositols (GPIs), leading to impairment of the parasite's ability to invade and replicate in the host cell. Surprisingly, attempts to rescue this defect through exogenous GlcNAc supplementation fail to completely restore these vital functions. In depth metabolomic analyses elucidate diverse causes underlying the failed rescue: utilization of GlcNAc is inefficient under glucose-replete conditions and fails to restore UDP-GlcNAc levels in GNA1-depleted parasites. In contrast, GlcNAc-supplementation under glucose-deplete conditions fully restores UDP-GlcNAc levels but fails to rescue the defects associated with GNA1 depletion. Our results underscore the importance of glucosamine-6-phosphate acetylation in governing T. gondii replication and invasion and highlight the potential of the evolutionary divergent GNA1 in Apicomplexa as a target for the development of much-needed new therapeutic strategies. Author summary: Toxoplasma gondii, Plasmodium, and Cryptosporidium spp., pose serious threats to human health. T. gondii, an intracellular and opportunistic pathogen, effectively avoids the host immune defences by forming long-lasting tissue cysts. Finding potent drugs to eliminate these persisting parasites remains a challenge. The glucosamine-phosphate-N-acetyltransferase (GNA1) catalyses a critical key step in the production of activated sugar nucleotides to build glycoconjugates essential for various functions in the cell. In P. falciparum, this enzyme has been identified as a potential target for antimalarial drugs. In this study, we explored the importance of this pathway in T. gondii and discovered that these sugar-containing compounds play a vital role in the parasite's ability to invade and replicate in host cells–crucial processes for its survival and ability to cause disease. Intriguingly, unlike some organisms that can bypass the pathway, T. gondii relies critically on glucosamine-6-phosphate acetylation. This reliance sheds light on the parasite's distinct metabolic properties and highlights the pathway's potential as a target for new therapeutic strategies. [ABSTRACT FROM AUTHOR]

Pyruvate lies at a pivotal node of carbon metabolism in eukaryotes. It is involved in diverse metabolic pathways in multiple organelles, and its interorganelle shuttling is crucial for cell fitness. Many apicomplexan parasites harbor a unique organelle called the apicoplast that houses metabolic pathways like fatty acid and isoprenoid precursor biosyntheses, requiring pyruvate as a substrate. However, how pyruvate is supplied in the apicoplast remains enigmatic. Here, deploying the zoonotic parasite Toxoplasma gondii as a model apicomplexan, we identified two proteins residing in the apicoplast membranes that together constitute a functional apicoplast pyruvate carrier (APC) to mediate the import of cytosolic pyruvate. Depletion of APC results in reduced activities of metabolic pathways in the apicoplast and impaired integrity of this organelle, leading to parasite growth arrest. APC is a pyruvate transporter in diverse apicomplexan parasites, suggesting a common strategy for pyruvate acquisition by the apicoplast in these clinically relevant intracellular pathogens. [ABSTRACT FROM AUTHOR]

The genus Borrelia has been divided into Borreliella spp., which can cause Lyme Disease (LD), and Borrelia spp., which can cause Relapsing Fever (RF). The distribution of genus Borrelia has broadened due to factors such as climate change, alterations in land use, and enhanced human and animal mobility. Consequently, there is an increasing necessity for a One Health strategy to identify the key components in the Borrelia transmission cycle by monitoring the human-animal-environment interactions. The aim of this study is to summarize all accessible data to increase our understanding and provide a comprehensive overview of Borrelia distribution in the Mediterranean region. Databases including PubMed, Google Scholar, and Google were searched to determine the presence of Borreliella and Borrelia spp. in vectors, animals, and humans in countries around the Mediterranean Sea. A total of 3026 were identified and screened and after exclusion of papers that did not fulfill the including criteria, 429 were used. After examination of the available literature, it was revealed that various species associated with LD and RF are prevalent in vectors, animals, and humans in Mediterranean countries and should be monitored in order to effectively manage and prevent potential infections. [ABSTRACT FROM AUTHOR]

Toxoplasma gondii is a foodborne pathogen that can cause severe and life-threatening infections in fetuses and immunocompromised patients. Felids are its only definitive hosts, and a wide range of animals, including humans, serve as intermediate hosts. When the transmissible bradyzoite stage is orally ingested by felids, they transform into merozoites that expand asexually, ultimately generating millions of gametes for the parasite sexual cycle. However, bradyzoites in intermediate hosts differentiate exclusively to disease-causing tachyzoites, which rapidly disseminate throughout the host. Though tachyzoites are well-studied, the molecular mechanisms governing transitioning between developmental stages are poorly understood. Each parasite stage can be distinguished by a characteristic transcriptional signature, with one signature being repressed during the other stages. Switching between stages require substantial changes in the proteome, which is achieved in part by ubiquitination. F-box proteins mediate protein poly-ubiquitination by recruiting substrates to SKP1, Cullin-1, F-Box protein E3 ubiquitin ligase (SCF-E3) complexes. We have identified an F-box protein named Toxoplasma gondii F-Box Protein L2 (TgFBXL2), which localizes to distinct perinucleolar sites. TgFBXL2 is stably engaged in an SCF-E3 complex that is surprisingly also associated with a COP9 signalosome complex that negatively regulates SCF-E3 function. At the cellular level, TgFBXL2-depleted parasites are severely defective in centrosome replication and daughter cell development. Most remarkable, RNAseq data show that TgFBXL2 conditional depletion induces the expression of stage-specific genes including a a large cohort of genes necessary for sexual commitment. Together, these data suggest that TgFBXL2 is a latent guardian of stage specific gene expression in Toxoplasma and poised to remove conflicting proteins in response to an unknown trigger of development. Author summary: Toxoplasma gondii exists as three major forms—disease-causing tachyzoites, transmissible bradyzoites, and sexual stages that develop in felid intestines. Each stage is typified by stage specific genes that are silenced in other stages and proper silencing is important for parasite growth. Previously, we identified in Toxoplasma a family of proteins, called F-box proteins, that are the substrate recruiting subunits of the SCF-E3 ubiquitin ligase complex. Of these, TgFBLX2 is predicted to be the most important for tachyzoite growth. Here, epitope-tagged TgFBLX2 was found to localize to a perinucleolar compartment in tachyzoites without apparently interacting with chromatin. A TgFBLX2 conditional expression strain revealed that TgFBLX2 is an essential tachyzoite gene whose loss leads to centrosome replication and daughter cell development defects. Next, we used RNAseq analysis to identify TgFBLX2-regulated genes and found that its loss led to the aberrant expression of non-tachyzoite stage specific genes including many sexual stage genes. Interestingly, TgFBLX2 interactome analyses revealed that the TgFBLX2-containing SCF-E3 is associated with the COP9 signalsome, which is a negative regulator of the SCF-E3 but does not interact with other known regulators of stage specific gene expression. Together, these data reveal TgFBLX2 as a novel negative regulator of stage specific gene expression. [ABSTRACT FROM AUTHOR]

Liquid‒liquid phase separation (LLPS) is a ubiquitous process in which proteins, RNA, and biomolecules assemble into membrane‐less compartments, playing important roles in many biological functions and diseases. The current knowledge on the biophysical and biochemical principles of LLPS is largely from in vitro studies; however, the physiological environment in living cells is complex and not at equilibrium. The characteristics of intracellular dynamics and their roles in physiological LLPS remain to be resolved. Here, by using single‐particle tracking of quantum dots and dynamic monitoring of the formation of stress granules (SGs) in single cells, the spatiotemporal dynamics of intracellular transport in cells undergoing LLPS are quantified. It is shown that intracellular diffusion and active transport are both reduced. Furthermore, the formation of SG droplets contributes to increased spatial heterogeneity within the cell. More importantly, the study demonstrated that the LLPS of SGs can be regulated by intracellular dynamics in two stages: the reduced intracellular diffusion promotes SG assembly and the microtubule‐associated transport facilitates SG coalescences. The work on intracellular dynamics not only improves the understanding of the mechanism of physiology phase separations occurring in nonequilibrium environments but also reveals an interplay between intracellular dynamics and LLPS. [ABSTRACT FROM AUTHOR]

We have previously shown that polygenic risk scores (PRS) can improve risk stratification of peripheral artery disease (PAD) in a large, retrospective cohort. Here, we evaluate the potential of PRS in improving the detection of PAD and prediction of major adverse cardiovascular and cerebrovascular events (MACCE) and adverse events (AE) in an institutional patient cohort. We created a cohort of 278 patients (52 cases and 226 controls) and fit a PAD-specific PRS based on the weighted sum of risk alleles. We built traditional clinical risk models and machine learning (ML) models using clinical and genetic variables to detect PAD, MACCE, and AE. The models' performances were measured using the area under the curve (AUC), net reclassification index (NRI), integrated discrimination improvement (IDI), and Brier score. We also evaluated the clinical utility of our PAD model using decision curve analysis (DCA). We found a modest, but not statistically significant improvement in the PAD detection model's performance with the inclusion of PRS from 0.902 (95% CI: 0.846–0.957) (clinical variables only) to 0.909 (95% CI: 0.856–0.961) (clinical variables with PRS). The PRS inclusion significantly improved risk re-classification of PAD with an NRI of 0.07 (95% CI: 0.002–0.137), p = 0.04. For our ML model predicting MACCE, the addition of PRS did not significantly improve the AUC, however, NRI analysis demonstrated significant improvement in risk re-classification (p = 2e-05). Decision curve analysis showed higher net benefit of our combined PRS-clinical model across all thresholds of PAD detection. Including PRS to a clinical PAD-risk model was associated with improvement in risk stratification and clinical utility, although we did not see a significant change in AUC. This result underscores the potential clinical utility of incorporating PRS data into clinical risk models for prevalent PAD and the need for use of evaluation metrics that can discern the clinical impact of using new biomarkers in smaller populations. [ABSTRACT FROM AUTHOR]

Toxoplasma gondii is an intracellular parasite that is important in medicine and veterinary science and undergoes distinct developmental transitions in its intermediate and definitive hosts. The switch between stages of T. gondii is meticulously regulated by a variety of factors. Previous studies have explored the role of the microrchidia (MORC) protein complex as a transcriptional suppressor of sexual commitment. By utilizing immunoprecipitation and mass spectrometry, constituents of this protein complex have been identified, including MORC, Histone Deacetylase 3 (HDAC3), and several ApiAP2 transcription factors. Conditional knockout of MORC or inhibition of HDAC3 results in upregulation of a set of genes associated with schizogony and sexual stages in T. gondii tachyzoites. Here, our focus extends to two primary ApiAP2s (AP2XII-1 and AP2XI-2), demonstrating their significant impact on the fitness of asexual tachyzoites and their target genes. Notably, the targeted disruption of AP2XII-1 and AP2XI-2 resulted in a profound alteration in merozoite-specific genes targeted by the MORC–HDAC3 complex. Additionally, considerable overlap was observed in downstream gene profiles between AP2XII-1 and AP2XI-2, with AP2XII-1 specifically binding to a subset of ApiAP2 transcription factors, including AP2XI-2. These findings reveal an intricate cascade of ApiAP2 regulatory networks involved in T. gondii schizogony development, orchestrated by AP2XII-1 and AP2XI-2. This study provides valuable insights into the transcriptional regulation of T. gondii growth and development, shedding light on the intricate life cycle of this parasitic pathogen. [ABSTRACT FROM AUTHOR]

Toxoplasma gondii resides in its intracellular niche by employing a series of specialized secretory organelles that play roles in invasion, host cell manipulation, and parasite replication. Rab GTPases are major regulators of the parasite's secretory traffic that function as nucleotide-dependent molecular switches to control vesicle trafficking. While many of the Rab proteins have been characterized in T. gondii, precisely how these Rabs are regulated remains poorly understood. To better understand the parasite's secretory traffic, we investigated the entire family of Tre2-Bub2-Cdc16 (TBC) domain-containing proteins, which are known to be involved in vesicle fusion and secretory protein trafficking. We first determined the localization of all 18 TBC domain-containing proteins to discrete regions of the secretory pathway or other vesicles in the parasite. Second, we use an auxin-inducible degron approach to demonstrate that the protozoan-specific TgTBC9 protein, which localizes to the endoplasmic reticulum (ER), is essential for parasite survival. Knockdown of TgTBC9 results in parasite growth arrest and affects the organization of the ER and mitochondrial morphology. TgTBC9 knockdown also results in the formation of large lipid droplets (LDs) and multi-membranous structures surrounded by ER membranes, further indicating a disruption of ER functions. We show that the conserved dual-finger active site in the TBC domain of the protein is critical for its GTPase-activating protein (GAP) function and that the Plasmodium falciparum orthologue of TgTBC9 can rescue the lethal knockdown. We additionally show by immunoprecipitation and yeast 2 hybrid analyses that TgTBC9 preferentially binds Rab2, indicating that the TBC9-Rab2 pair controls ER morphology and vesicular trafficking in the parasite. Together, these studies identify the first essential TBC protein described in any protozoan and provide new insight into intracellular vesicle trafficking in T. gondii. Toxoplasma gondii regulates vesicle trafficking by secreting Rab GTPases from specialised organelles. This study characterizes all Tre2-Bub2-Cdc16 (TBC) domain-containing proteins in T. gondii, identifying TBC9 as an essential regulator of Rab2 that controls intracellular vesicular trafficking. [ABSTRACT FROM AUTHOR]

Variable taxi time prediction is the core of the Airport Collaborative Decision Making (A-CDM) system. An accurate taxi time prediction contributes to enhancing airport operational efficiency, safety and predictability. The deep dynamic spatio-temporal correlation inherent in airport traffic data is critical for taxi time prediction. However, existing machine learning (deep learning) methods have been unable to thoroughly exploit these correlations. To address this issue, we propose a deep learning-based model called the multi-task dynamic spatio-temporal graph attention network (MT-DSTGAN). Our model also predicts future entire airport traffic flow and taxiing segment traffic flow as auxiliary tasks, with the goal of enhancing the accuracy of aircrafts' taxi time prediction. The proposed MT-DSTGAN model is implemented and assessed through a case study of Beijing Capital International Airport with a real-world dataset. The advantage of the proposed model, which shows better performance in various evaluation metrics, is demonstrated in a comparative study with other baseline works. In summary, the proposed MT-DSTGAN exhibits promising capabilities in perceiving the dynamic changes in the taxiing process of aircraft and demonstrates the ability to capture complex spatio-temporal correlations in airport traffic data. [ABSTRACT FROM AUTHOR]

Water electrolysis is considered the most promising technology for hydrogen production. In the present study, RCoFe (regulated cobalt ferricyanide) is synthesized via the co-precipitation method, which is characterized by physical and electrochemical properties. XRD confirmed the crystalline phase of RCoFe and the diffraction peaks are consistent with JCPDS#82-2284. FE-SEM examines the nanoplatelet morphology, which could help with the mass transfer of electrons during the electrochemical process. Here, RCoFe promotes the extensive activity for the OER (oxygen evolution reaction) with 270 mV/GC; 252 mV/NF at 10 mA cm−2. The lower Tafel value for RCoFe (78 mV dec−1/GC; 107 mV dec−1/NF) was derived from LSV when compared to IrO2 (98 mV dec−1/GC; 123 mV dec−1/NF) and stable for a long time with minimal loss of potential (130 h/GC-1.5%; 250 h/NF-1.8%) owing to citrus sap, which stabilized the electrocatalyst. After electrochemical treatment, the post-SEM morphology reveals that the nanoplatelets are completely bound together to enhance the OER process. [ABSTRACT FROM AUTHOR]

Recombinant proteins are gaining increasing popularity for treating human diseases. The clinical effectiveness of recombinant proteins is directly related to their biological activity, which is an important indicator in drug development and quality control. However, certain recombinant proteins have unclear or complex signal pathways, making detecting their activity in vitro difficult. For instance, recombinant human endostatin (endostatin), a new antitumor drug developed in China, lacks a sensitive and stable assay for its biological activity since being market approval. To address this issue, we performed a genome‐wide screening of immortalized human umbilical vein endothelial cells (HUVECs) using a CRISPR/Cas9 knockout library containing 20,000 targeted genes. We identified two potential endostatin‐resistant genes, NEPSPP and UTS2, and successfully constructed a highly sensitive cell line, HUVEC‐UTS2‐3#, by knocking down the UTS2 gene. Based on the optimized parameters of HUVEC‐UTS2‐3# cells, we established a new method for detecting the biological activity of endostatin. The method was validated, and it produced results consistent with primary HUVEC cells but with higher sensitivity and more stable data. The use of gene‐editing technology provides a novel solution for detecting the biological activity of recombinant proteins that other methods cannot detect. [ABSTRACT FROM AUTHOR]

Simple Summary: Ammonia is a major water quality factor influencing the survival and health of shrimp, among which the gill is the main effector organ for ammonia toxicity. In this study, we explored the toxicity of ammonia stress on the physiological homeostasis in the gills of Litopenaeus vannamei under seawater and low-salinity conditions. This study included four groups, namely the SC group (ammonia-N 0 mg/L, salinity 30‰), SAN group (ammonia-N 10 mg/L, salinity 30‰), LC group (ammonia-N 0 mg/L, salinity 3‰), and LAN group (ammonia-N 10 mg/L, salinity 3‰). The ammonia stress lasted for 14 days. The results show that ammonia stress caused the severe contraction of gill filaments and the deformation or even rupture of gill vessels. Ammonia stress could also influence the redox, ER function, apoptosis, detoxification, energy metabolism, and osmoregulation of the shrimp gills. These results are helpful to analyze the toxicological mechanism of ammonia stress on the seawater- and low salinity-cultured shrimp. Ammonia is a major water quality factor influencing the survival and health of shrimp, among which the gill is the main effector organ for ammonia toxicity. In this study, we chose two types of Litopenaeus vannamei that were cultured in 30‰ seawater and domesticated in 3‰ low salinity, respectively, and then separately subjected to ammonia stress for 14 days under seawater and low-salinity conditions, of which the 3‰ low salinity-cultured shrimp were domesticated from the shrimp cultured in 30‰ seawater after 27 days of gradual salinity desalination. In detail, this study included four groups, namely the SC group (ammonia-N 0 mg/L, salinity 30‰), SAN group (ammonia-N 10 mg/L, salinity 30‰), LC group (ammonia-N 0 mg/L, salinity 3‰), and LAN group (ammonia-N 10 mg/L, salinity 3‰). The ammonia stress lasted for 14 days, and then the changes in the morphological structure and physiological function of the gills were explored. The results show that ammonia stress caused the severe contraction of gill filaments and the deformation or even rupture of gill vessels. Biochemical indicators of oxidative stress, including LPO and MDA contents, as well as T-AOC and GST activities, were increased in the SAN and LAN groups, while the activities of CAT and POD and the mRNA expression levels of antioxidant-related genes (nrf2, cat, gpx, hsp70, and trx) were decreased. In addition, the mRNA expression levels of the genes involved in ER stress (ire1 and xbp1), apoptosis (casp-3, casp-9, and jnk), detoxification (gst, ugt, and sult), glucose metabolism (pdh, hk, pk, and ldh), and the tricarboxylic acid cycle (mdh, cs, idh, and odh) were decreased in the SAN and LAN groups; the levels of electron-transport chain-related genes (ndh, cco, and coi), and the bip and sdh genes were decreased in the SAN group but increased in the LAN group; and the level of the ATPase gene was decreased but the cytc gene was increased in the SAN and LAN groups. The mRNA expression levels of osmotic regulation-related genes (nka-β, ca, aqp and clc) were decreased in the SAN group, while the level of the ca gene was increased in the LAN group; the nka-α gene was decreased in both two groups. The results demonstrate that ammonia stress could influence the physiological homeostasis of the shrimp gills, possibly by damaging the tissue morphology, and affecting the redox, ER function, apoptosis, detoxification, energy metabolism, and osmoregulation. [ABSTRACT FROM AUTHOR]

Here, we described the prevalence of Borrelia burgdorferi s.l. and Babesia species found in mono- and double infections among Ixodes ricinus ticks occurring in urban areas of the city of Poznań, Poland. We tested 1029 host-seeking ticks and 1268 engorged ticks removed from pet animals. Borrelia afzelii and B. garinii prevailed both in ticks from vegetation (3.7% and 3.7%, respectively) and from pets (3.7% and 0.6%, respectively). Babesia canis and Ba. microti were the most prevalent in host-seeking (2.6% and 1.4%, respectively) and feeding ticks (2.8% and 2.2%, respectively). Babesia microti sequences proved to be identical to the human pathogenic Ba. microti genotype "Jena/Germany". Sequences of the rarest piroplasm Ba. venatorum (0.7%) were identical with those isolated from European patients. About 1.0% of tested ticks yielded dual infections; in host-seeking ticks, Ba. canis prevailed in co-infections with B. afzelii and B. garinii, whereas Ba. microti and B. afzelii dominated in double-infected feeding ticks. Dual infections, even with a low prevalence, pose a challenge for differential diagnosis in patients with acute febrile disease after a tick bite. The finding of Ba. canis in both tick groups suggests that I. ricinus could be involved in the circulation of this piroplasm. [ABSTRACT FROM AUTHOR]

Toxoplasmosis continues to be a prevalent parasitic zoonosis with a global distribution. This disease is caused by an intracellular parasite known as Toxoplasma gondii, and the development of effective novel drug targets to combat it is imperative. There is limited information available on the potential advantages of wheat germ oil (WGO) and propolis, both individually and in combination, against the acute phase of toxoplasmosis. In this study, acute toxoplasmosis was induced in Swiss albino mice, followed by the treatment of infected animals with WGO and propolis, either separately or in combination. After 10 days of experimental infection and treatment, mice from all groups were sacrificed, and their brains, uteri, and kidneys were excised for histopathological assessment. Additionally, the average parasite load in the brain was determined through parasitological assessment, and quantification of the parasite was performed using Real-Time Polymerase Chain Reaction targeting gene amplification. Remarkably, the study found that treating infected animals with wheat germ oil and propolis significantly reduced the parasite load compared to the control group that was infected but not treated. Moreover, the group treated with a combination of wheat germ oil and propolis exhibited a markedly greater reduction in parasitic load compared to the other groups. Similarly, the combination treatment effectively restored the histopathological changes observed in the brain, uterus, and kidney, and the scoring of these reported lesions confirmed these findings. In summary, the present results reveal intriguing insights into the potential therapeutic benefits of wheat germ oil and propolis in the treatment of acute toxoplasmosis. [ABSTRACT FROM AUTHOR]

Aims: Frequent and extensive within-day glycemic variability (GV) in blood glucose levels may increase the risk of hypoglycemia and long-term mortality in hospitalized patients with diabetes. We aimed to assess the amplitude and frequency of within-day GV in inpatients with type 2 diabetes and to explore the factors influencing within-day GV. Methods: We conducted a single-center, retrospective observational study by analyzing hospital records and 10-day real-time continuous glucose monitoring data. Within-day GV was assessed using the coefficient of variation (%CV). The primary outcome was the amplitude and frequency of within-day GV. The frequency of within-day GV was assessed by the consecutive days (CD) of maintaining within the target %CV range after first reaching it (CD after first reaching the target) and the maximum consecutive days of maintaining within the target %CV range (Max-CD). The target %CV range was less than 24.4%. We evaluated the factors influencing within-day GV using COX regression and Poisson regression models. Results: A total of 1050 cases were analyzed, of whom 86.57% reduced the amplitude of within-day GV before the sixth day of hospitalization. Of the 1050 hospitalized patients, 66.57% stayed within the target %CV range for less than two days after first reaching the target and 69.71% experienced a Max-CD of fewer than four days. Reducing the average postprandial glucose excursion (hazard ratio [HR]: 0.81, 95% confidence interval [CI]: 0.77–0.85; incidence rate ratios [IRR]: 0.72, 95% CI: 0.69–0.74) and the use of α-glucosidase inhibitors (IRR: 1.1, 95% CI: 1.01–1.18) and glucagon-like peptide-1 agonist (IRR: 1.30, 95% CI: 1.02–1.65) contributed to reducing the amplitude and decreasing the frequency of within-day GV. However, the use of insulin (HR: 0.64, 95% CI: 0.55–0.75; IRR: 0.86, 95% CI: 0.79–0.93) and glinide (HR: 0.47, 95% CI: 0.31–0.73; IRR: 0.84, 95% CI: 0.73–0.97) may lead to an increased frequency of within-day GV. Conclusions: An increasing frequency of within-day GV was observed during the hospitalization in patients with type 2 diabetes, despite the effective reduction in the amplitude of within-day GV. Using medications designed to lower postprandial blood glucose could contribute to minimize the risk of frequent within-day GV. [ABSTRACT FROM AUTHOR]

Background: Genome manipulation of Leishmania species and the creation of modified strains are widely employed strategies for various purposes, including gene function studies, the development of live attenuated vaccines, and the engineering of host cells for protein production. Objective: Despite the introduction of novel manipulation approaches like CRISPR/Cas9 technology with significant advancements in recent years, the development of a reliable protocol for efficiently and precisely altering the genes of Leishmania strains remains a challenging endeavor. Following the successful adaptation of the CRISPR/Cas9 system for higher eukaryotic cells, several research groups have endeavored to apply this system to manipulate the genome of Leishmania. Results: Despite the substantial differences between Leishmania and higher eukaryotes, the CRISPR/Cas9 system has been effectively tested and applied in Leishmania. Conclusion: This comprehensive review summarizes all the CRISPR/Cas9 systems that have been employed in Leishmania, providing details on their methods and the expression systems for Cas9 and gRNA. The review also explores the various applications of the CRISPR system in Leishmania, including the deletion of multicopy gene families, the development of the Leishmania vaccine, complete gene deletions, investigations into chromosomal translocations, protein tagging, gene replacement, large-scale gene knockout, genome editing through cytosine base replacement, and its innovative use in the detection of Leishmania. In addition, the review offers an up-to-date overview of all double-strand break repair mechanisms in Leishmania. [ABSTRACT FROM AUTHOR]

With increasing demands on the quality and quantity of produced biomass, as the main element of the knowledge-based economy, people and the issue of safety and health protection at work are coming to the fore. The aim of the work is the synthesis and overview of the results of the analysis of the health and safety risks of the production of forest biomass in various production phases, starting with its cultivation, through the harvesting production and transport process, up to the issue of its safe storage until it is used for the production of primary energy. Based on the analyzed overview of the existing risks in the production and storage of biomass, it can be concluded that the largest number of works is dedicated to the technological process of storage and consumption of the produced forms of biomass. Of the risks in this phase, the largest number of works is devoted to the risks of the production of spores of phytopathogens and fungi threatening human health. Further research should be primarily oriented toward creating models and modeling the processes of the emergence of these risk factors and the dynamics of their growth. [ABSTRACT FROM AUTHOR]

The food insulin index (FII) is a novel algorithm used to determine insulin responses of carbohydrates, proteins, and fats. This scoping review aimed to provide an overview of all scientifically relevant information presented on the application of the FII in the prevention and management of insulin resistance and diabetes. The Arksey and O'Malley framework and the PRISMA Extension for Scoping Reviews 22-item checklist were used to ensure that all areas were covered in the scoping review. Our search identified 394 articles, of which 25 articles were included. Three main themes emerged from the included articles: 1. the association of FII with the development of metabolic syndrome, insulin resistance, and diabetes, 2. the comparison of FII with carbohydrate counting (CC) for the prediction of postprandial insulin response, and 3. the effect of metabolic status on the FII. Studies indicated that the FII can predict postprandial insulin response more accurately than CC, and that a high DII and DIL diet is associated with the development of metabolic syndrome, insulin resistance, and diabetes. The FII could be a valuable tool to use in the prevention and management of T1DM, insulin resistance, and T2DM, but more research is needed in this field. [ABSTRACT FROM AUTHOR]

The success of the intracellular parasite Toxoplasma gondii in invading host cells relies on the apical complex, a specialized microtubule cytoskeleton structure associated with secretory organelles. The T. gondii genome encodes three isoforms of both α- and β-tubulin, which undergo specific post-translational modifications (PTMs), altering the biochemical and biophysical proprieties of microtubules and modulating their interaction with associated proteins. Tubulin PTMs represent a powerful and evolutionarily conserved mechanism for generating tubulin diversity, forming a biochemical 'tubulin code' interpretable by microtubule-interacting factors. T. gondii exhibits various tubulin PTMs, including α-tubulin acetylation, α-tubulin detyrosination, Δ5α-tubulin, Δ2α-tubulin, α- and β-tubulin polyglutamylation, and α- and β -tubulin methylation. Tubulin glutamylation emerges as a key player in microtubule remodeling in Toxoplasma, regulating stability, dynamics, interaction with motor proteins, and severing enzymes. The balance of tubulin glutamylation is maintained through the coordinated action of polyglutamylases and deglutamylating enzymes. This work reviews and discusses current knowledge on T. gondii tubulin glutamylation. Through in silico identification of protein orthologs, we update the recognition of putative proteins related to glutamylation, contributing to a deeper understanding of its role in T. gondii biology. [ABSTRACT FROM AUTHOR]

Euglena gracilis, a model organism of the eukaryotic supergroup Discoba harbouring also clinically important parasitic species, possesses diverse metabolic strategies and an atypical electron transport chain. While structures of the electron transport chain complexes and supercomplexes of most other eukaryotic clades have been reported, no similar structure is currently available for Discoba, limiting the understandings of its core metabolism and leaving a gap in the evolutionary tree of eukaryotic bioenergetics. Here, we report high-resolution cryo-EM structures of Euglena's respirasome I + III2 + IV and supercomplex III2 + IV2. A previously unreported fatty acid synthesis domain locates on the tip of complex I's peripheral arm, providing a clear picture of its atypical subunit composition identified previously. Individual complexes are re-arranged in the respirasome to adapt to the non-uniform membrane curvature of the discoidal cristae. Furthermore, Euglena's conformationally rigid complex I is deactivated by restricting ubiquinone's access to its substrate tunnel. Our findings provide structural insights for therapeutic developments against euglenozoan parasite infections. Euglena gracilis is a model organism of the eukaryotic supergroup Discoba, single-celled organisms containing mitochondria with discoid cristae. Here, the authors report structures of Euglena's transport chain supercomplex to reveal their distinctive architecture and working mechanisms. [ABSTRACT FROM AUTHOR]

Airport arrival and departure movements are characterized by high dynamism, stochasticity, and uncertainty. Therefore, it is of paramount importance to predict and analyze surface taxi time accurately and scientifically. This paper conducts a comprehensive review of existing studies on surface taxi time prediction and analysis. Firstly, the overall research framework of surface taxi time prediction and analysis is categorized from three perspectives: taxi time type, movement type, and modeling method. Then, focusing on the two means of taxi time analytical modeling and simulation modeling, the existing mainstream models and methods are categorized, and the main ideas and scope of application of the various methods are analyzed. Finally, the paper presents the future development direction of surface taxi time prediction prospects. The research results are aimed at providing basic support and methodological guidance for reducing the uncertainty in airport surface operation and enhancing the level of control and decision-making ability of airport surface operation. [ABSTRACT FROM AUTHOR]

Background: Essential genes encode functions that play a vital role in the life activities of organisms, encompassing growth, development, immune system functioning, and cell structure maintenance. Conventional experimental techniques for identifying essential genes are resource-intensive and time-consuming, and the accuracy of current machine learning models needs further enhancement. Therefore, it is crucial to develop a robust computational model to accurately predict essential genes. Results: In this study, we introduce GCNN-SFM, a computational model for identifying essential genes in organisms, based on graph convolutional neural networks (GCNN). GCNN-SFM integrates a graph convolutional layer, a convolutional layer, and a fully connected layer to model and extract features from gene sequences of essential genes. Initially, the gene sequence is transformed into a feature map using coding techniques. Subsequently, a multi-layer GCN is employed to perform graph convolution operations, effectively capturing both local and global features of the gene sequence. Further feature extraction is performed, followed by integrating convolution and fully-connected layers to generate prediction results for essential genes. The gradient descent algorithm is utilized to iteratively update the cross-entropy loss function, thereby enhancing the accuracy of the prediction results. Meanwhile, model parameters are tuned to determine the optimal parameter combination that yields the best prediction performance during training. Conclusions: Experimental evaluation demonstrates that GCNN-SFM surpasses various advanced essential gene prediction models and achieves an average accuracy of 94.53%. This study presents a novel and effective approach for identifying essential genes, which has significant implications for biology and genomics research. [ABSTRACT FROM AUTHOR]

Ixodes ricinus nymphs and adults removed from humans, and larvae and nymphs from birds, have been analysed for infection with Babesia species and Borrelia species previously in separately published studies. Here, we use the same data set to explore the coinfection pattern of Babesia and Borrelia species in the ticks. We also provide an overview of the ecology and potential public health importance in Sweden of I. ricinus infected both with zoonotic Babesia and Borrelia species. Among 1952 nymphs and adult ticks removed from humans, 3.1% were PCR-positive for Babesia spp. Of these Babesia-positive ticks, 43% were simultaneously Borrelia-positive. Among 1046 immatures of I. ricinus removed from birds, 2.5% were Babesia-positive, of which 38% were coinfected with Borrelia species. This study shows that in I. ricinus infesting humans or birds in Sweden, potentially zoonotic Babesia protozoa sometimes co-occur with human-pathogenic Borrelia spp. Diagnostic tests for Babesia spp. infection are rarely performed in Europe, and the medical significance of this pathogen in Europe could be underestimated. [ABSTRACT FROM AUTHOR]

Examining host-pathogen interactions in animals can capture aspects of infection that are obscured in cell culture. Using CRISPR-based screens, we functionally profile the entire genome of the apicomplexan parasite Toxoplasma gondii during murine infection. Barcoded gRNAs enabled bottleneck detection and mapping of population structures within parasite lineages. Over 300 genes with previously unknown roles in infection were found to modulate parasite fitness in mice. Candidates span multiple axes of host-parasite interaction. Rhoptry Apical Surface Protein 1 was characterized as a mediator of host-cell tropism that facilitates repeated invasion attempts. GTP cyclohydrolase I was also required for fitness in mice and druggable through a repurposed compound, 2,4-diamino-6-hydroxypyrimidine. This compound synergized with pyrimethamine against T. gondii and malaria-causing Plasmodium falciparum parasites. This work represents a complete survey of an apicomplexan genome during infection of an animal host and points to novel interfaces of host-parasite interaction., (© 2024. The Author(s), under exclusive licence to Springer Nature Limited.)

Mastering electrodes' formulations is a complex and tedious task, because for each composition of electroactive material(s) it is necessary to adjust the inactive additives nature and content to optimize battery performance. In this direction, the amount of binder is proposed to be adjusted to the surface developed by all of the powders involved in the composition of the electrode, i.e., the electroactive materials and electronic conductive additives. This concept, introduces here as binder-to-powders coverage ratio, relies upon the micromechanical models developed in the field of polymer-based composite materials. The validity of this new electrode formulation parameter is shown here for two different SiO x /Graphite blends, which differ in the type of graphite, and for blends of two different binders, polyacrylic acid and styrene-butadiene rubber. At the optimal coverage ratio, a satisfactory capacity retention is obtained in full cell with an ethylene carbonate free and fluoroethylene carbonate rich electrolyte., (© 2023 Wiley‐VCH GmbH.)

Calixarenes are displaying great potential for the development of new drug delivery systems, diagnostic imaging, biosensing devices and inhibitors of biological processes. In particular, calixarene derivatives are able to interact with many different enzymes and function as inhibitors. By screening of the potential drug target database (PDTD) with a reverse docking procedure, we identify and discuss a selection of 100 proteins that interact strongly with calix[4]arene. We also discover that leucine (23.5 %), isoleucine (11.3 %), phenylalanines (11.3 %) and valine (9.5 %) are the most frequent binding residues followed by hydrophobic cysteines and methionines and aromatic histidines, tyrosines and tryptophanes. Top binders are peroxisome proliferator-activated receptors that already are targeted by commercial drugs, demonstrating the practical interest in calix[4]arene. Nuclear receptors, potassium channel, several carrier proteins, a variety of cancer-related proteins and viral proteins are prominent in the list. It is concluded that calix[4]arene, which is characterized by facile access, well-defined conformational characteristics, and ease of functionalization at both the lower and higher rims, could be a potential lead compound for the development of enzyme inhibitors and theranostic platforms., (© 2024 Wiley-VCH GmbH.)

Cytokinetic abscission marks the final stage of cell division, during which the daughter cells physically separate through the generation of new barriers, such as the plasma membrane or cell wall. While the contractile ring plays a central role during cytokinesis in bacteria, fungi and animal cells, the process diverges in Apicomplexa. In Toxoplasma gondii, two daughter cells are formed within the mother cell by endodyogeny. The mechanism by which the progeny cells acquire their plasma membrane during the disassembly of the mother cell, allowing daughter cells to emerge, remains unknown. Here we identify and characterize five T. gondii proteins, including three protein phosphatase 2A subunits, which exhibit a distinct and dynamic localization pattern during parasite division. Individual downregulation of these proteins prevents the accumulation of plasma membrane at the division plane, preventing the completion of cellular abscission. Remarkably, the absence of cytokinetic abscission does not hinder the completion of subsequent division cycles. The resulting progeny are able to egress from the infected cells but fail to glide and invade, except in cases of conjoined twin parasites., (© 2024. The Author(s).)

Background and Aims: Lyme disease is a well-known occupational risk across North America caused by exposure to Borrelia burgdorferi via blacklegged ticks (Ixodes scapularis). As the geographic range of B. burgdorferi advances with the increasing distribution of blacklegged ticks, more outdoor workers are at risk of contracting Lyme disease. In this study, we examined the demography and personal perceptions of outdoor workers within one framework to better determine the overall risk for those working outdoors., Methods and Results: We analysed outdoor worker knowledge of ticks and of behaviours that can prevent tick bites and Lyme disease. We then compared these risk perceptions of individuals across age, sex, education, and industry, as well as time spent outdoors. We tested the hypothesis that the risk perception of an individual and their knowledge of Lyme disease transmission was dependent on their demographics, experience in their job, and the region in which they spend time outdoors. We estimated a knowledge-based risk score based on individuals' answers to a questionnaire on risk perception given to voluntary participants who work outdoors. Those who had higher risk scores were more at risk. We found that knowledge-based risk scores were correlated with geographic risk levels and with the number of hours per day spent outdoors. Those who work longer hours and who work in areas with mid-level risk had higher risk scores. Those who spend more time outdoors recreationally had lower risk scores., Conclusions: Further examination and acknowledgment of the reasoning behind why these factors are affecting workers' risks must be considered to recognize that it is not necessarily demographics or geographically high-risk areas that affect an individual's risk. Workers' knowledge of these risks is affected by several variables that should be taken into consideration when implementing safety measures and awareness programs., (© 2024 The Author(s). Zoonoses and Public Health published by Wiley‐VCH GmbH.)

Globally sustainable disease management ensuring high quality in grapes is in demand as it holds significant importance as a versatile fruit for consumption, winemaking, and production of various products such as grape juice, raisin, and grape-seed oil. The present paper reports a combination of nano-biotechnology as a promising strategy for enhancing plant health and fruit productivity in grapes combining Irradiated chitosan nanoparticles and bio-control agents. The Irradiated Chitosan with Bacillus subtilis and Trichoderma viridae and pesticides were evaluated for disease management. Percent disease index, percent disease control, and percent yield enhancement in Cymoxanil 8% + Mamcozeb 64% WP @ 0.2% treatment were as 17. 24%, 67.97% and 33.91% in 150 ppm Irradiated chitosan+B. subtilis were 19.83, 63.16, 30.41 and in Trichoderma 150 ppm Irradiated chitosan were 24.58, 54.33, and 27.40, respectively as compared to untreated crop with disease severity 53.84% PDI. Thus, irradiated chitosan and Bacillus subtilis elucidated a synergistic combination for residue-free efficient phytosanitary measures, which harnessed the strength of chitosan and bio-control agents for sustainable grape productivity. These findings will also pave the way for a deeper understanding of the synergistic interaction between Irradiated nanochitosan and bio-control agents for an eco-friendly and economically viable disease management strategy. The minimum temperature and morning relative humidity (RH I) had positive significance, with correlation coefficients of 0.484 and 0.485, respectively. The evening relative humidity (RH II) had a positive highly significant positive correlation coefficient of 0.664. Chitosan merits as a multiple stress tolerance enhancing agent that will further help in mitigating climate change adaptations in grapevines reducing reliance on chemical agro-inputs., (© 2024 Wiley Periodicals LLC.)

Background: Diabetes is a complex health condition requiring medical therapy and lifestyle modifications to attain treatment targets. Previous studies have not fully explored factors associated with adherence to medication, diets and physical activity recommendations among individuals living with diabetes in rural South Africa. We examined the association between knowledge, health belief and adherence to medication, dietary, and physical activity recommendations and explored self-reported reasons for non-adherence., Methods: This cross-sectional study was conducted among 399 individuals living with diabetes recruited over 12 weeks from six randomly selected primary healthcare centres in rural South Africa. Sociodemographic and clinical data were obtained by self-report. Health beliefs, knowledge, and adherence were assessed using validated measures. Descriptive and inferential statistics were carried out., Results: The majority (81.7%) of the participants were females, with a mean age of 62 ± 11 years. Only 39% reported adhering to their prescribed medication regimen, 25% reported adhering to dietary recommendations, and 32% reported adhering to physical activity recommendations. The most cited reasons for non-adherence were lack of access to (n = 64) and cost of drugs (n = 50), perceived high costs of healthy diets (n = 243), and lack of time (n = 181) for physical activity. Level of education was an independent predictor of medication adherence [odds ratio, OR = 2.02 (95% confidence interval, CI: 1.20-3.40)] while diabetes knowledge was independently associated with both medications [OR = 3.04 (95% CI: 1.78-45.12)]; and physical activity adherence [OR = 2.92 (95% CI: 1.04-2.96). Positive health belief was independently associated with adherence to medications [OR = 1.72 (95% CI: 1.15-2.57) and dietary recommendations [OR = 1.75 (95% CI: 1.04-2.96)]., Conclusion: Adherence to three important self-care practices, medication, diet, and physical activity, was suboptimal in this study setting. Socioeconomic reasons and access barriers were significant drivers of non-adherence, while increased knowledge and positive health beliefs were potential facilitators. Efforts to improve medication adherence and foster engagement in healthy lifestyle behaviours must consider patients' knowledge and health beliefs. Primary healthcare providers should create awareness on the importance of adherence on health outcomes for people with diabetes. Likewise, efforts to increase the availability and affordability of medications for socioeconomically disadvantaged populations should be prioritised by the key health stakeholders., (© 2024 The Author(s). Journal of Evaluation in Clinical Practice published by John Wiley & Sons Ltd.)

With the growth of optogenetic research, the demand for optical probes tailored to specific applications is ever rising. Specifically, for applications like the coiled cochlea of the inner ear, where planar, stiff, and nonconformable probes can hardly be used, transitioning from commonly used stiff glass fibers to flexible probes is required, especially for long-term use. Following this demand, polydimethylsiloxane (PDMS) with its lower Young's modulus compared to glass fibers can serve as material of choice. Hence, the long-term usability of PDMS as a waveguide material with respect to variations in transmission and refractive index over time is investigated. Different manufacturing methods for PDMS-based flexible waveguides are established and compared with the aim to minimize optical losses and thus maximize optical output power. Finally, the waveguides with lowest optical losses (-4.8 dB cm -1 ± 1.3 dB cm -1 at 472 nm) are successfully inserted into the optogenetically modified cochlea of a Mongolian gerbil (Meriones unguiculatus), where optical stimuli delivered by the waveguides evoked robust neuronal responses in the auditory pathway., (© 2024 The Authors. Advanced Healthcare Materials published by Wiley‐VCH GmbH.)

Apicomplexan parasites are aetiological agents of numerous diseases in humans and livestock. Functional genomics studies in these parasites enable the identification of biological mechanisms and protein functions that can be targeted for therapeutic intervention. Recent improvements in forward genetics and whole-genome screens utilising CRISPR/Cas technology have revolutionised the functional analysis of genes during Apicomplexan infection of host cells. Here, we highlight key discoveries from CRISPR/Cas9 screens in Apicomplexa or their infected host cells and discuss remaining challenges to maximise this technology that may help answer fundamental questions about parasite-host interactions., (© 2024 The Authors. Molecular Microbiology published by John Wiley & Sons Ltd.)

The cochlea forms a key element of the human auditory system in the temporal bone. Damage to the cochlea continues to produce significant impairment for sensory reception of environmental stimuli. To improve this impairment, the optical cochlear implant forms a new research approach. A prerequisite for this method is to understand how light propagation, as well as scattering, reflection, and absorption, takes place within the cochlea. We offer a method to study the light distribution in the human cochlea through phantom materials which have the objective to mimic the optical behavior of bone and Monte-Carlo simulations. The calculation of an angular distribution after scattering requires a phase function. Often approximate functions like Henyey-Greenstein, two-term Henyey-Greenstein or Legendre polynomial decompositions are used as phase function. An alternative is to exactly calculate a Mie distribution for each scattering event. This method provides a better fit to the data measured in this work., (© 2023 The Authors. Journal of Biophotonics published by Wiley‐VCH GmbH.)

Fig wasps (Agaonidae; Hymenoptera) are the only pollinating insects of fig trees (Ficus; Moraceae), forming the most closely and highly specific mutualism with the host. We used transcriptome sequences of 25 fig wasps from six genera to explore the evolution of key molecular components of fig wasp chemosensory genes: odorant-binding proteins (OBPs) and chemosensory proteins (CSPs). We identified a total 321 OBPs and 240 CSPs, with each species recording from 6 to 27 OBP genes and 6-19 CSP genes. 318 OBP genes are clustered into 17 orthologous groups and can be divided into two groups: PBP sensitive to pheromone and GOBP sensitive to general odor molecules, such as alcohols, esters, acids, ketones, and terpenoids. 240 CSP genes are clustered into 12 orthologous groups, which can be divided into three major groups and have functions, such as olfactory, tissue formation and/or regeneration, developmental, and some specific and unknown function. The gene sequences of most orthologous groups vary greatly among species and are consistent with the phylogenetic relationships between fig wasps. Strong purifying selection of both OBP and CSP genes was detected, as shown by low ω values. A positive selection was detected in one locus in CSP1. In conclusion, the evolution of chemosensory proteins OBPs and CSPs in fig wasps is relatively conservative, and they play an indispensable role in the life activities of fig wasps. Our results provide a starting point for understanding the molecular basis of the chemosensory systems of fig wasps., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

The Eukaryotic Pathogen, Vector and Host Informatics Resources ( VEuPathDB.org ) provide free online access to omic data from eukaryotic protozoan and fungal pathogens, arthropod vectors of disease, and host responses to pathogen infection. The goal of VEuPathDB is to make data easily accessible, findable, and importantly, re-usable by laboratory scientists. All integrated data and analyses follow standard workflows and methods to ensure data accuracy and enable data interoperability. Integrated data include genomes and annotation, transcriptomic (e.g., single-cell/bulk RNA-sequence and microarray data), proteomic (e.g., mass spectrometry evidence and quantitative data), isolate sequencing data used for variant calling and copy number variation determination, epigenomics, whole-genome phenotyping data (e.g., CRISPR screens and large-scale imaging and subcellular localization data), etc. Standard analyses provide additional data such as InterPro domains, signal peptide and transmembrane domain predictions, and metabolic pathways. Comparative genomic analysis in VEuPathDB is facilitated by leveraging orthology to enable the transformation of results between organisms and identifying genes with specific phyletic patterns. In addition, synteny between genomes is facilitated by shading orthologs across species and strains. Accessibility to and re-usability of the data is made possible through specialized searches and a graphical search strategy system that enables scientists to build in silico experiments combining results from multiple experiments with diverse data types., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)