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Summary: Here, we present a protocol for encapsulating DNA molecules under crowded conditions within cell-sized lipid-coated droplets. We describe steps for preparing a lipid-oil mixture and adding an aqueous solution containing DNA, which, when mixed, forms water-in-oil droplets of radii between ∼5 and 100 μm. We then detail procedures for quantifying the dynamics of DNA molecules in these droplets by analyzing fluorescence microscopy time series using differential dynamic microscopy. This protocol can be utilized to investigate DNA transport within a range of biomimetic and crowded environments.For complete details on the use and execution of this protocol, please refer to Aporvari et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Helminth parasite infections are widespread in smallholder farming systems affecting farmers and livestock animals. There are pathogenic parasites that populate the gut of their host and coexist closely with the gut microbiota. The physical and immunological environment of the gut can be modified by parasites and microbiota creating a wide range of interactions. These interactions modify the development of infection, affects overall host health, and can modify the way a host interacts with its bacterial microbiota. In addition, where there is a high worm burden parasites will affect the health of the host and intestinal tract colonization. This review highlights key studies on the interaction between helminth parasites and the intestinal microbiome to understand the relationship between parasitic worm infections and gut microbiome health in chickens. Finally, the review discusses modulations, molecular changes, and the importance of helminth-microbiome interactions for the host. [ABSTRACT FROM AUTHOR]

Abstract Susceptibility of human cells to cold stress restricts the use of therapeutic hypothermia and long-term preservation of organs at low temperatures. In contrast, cells of mammalian hibernators possess remarkable cold resistance, but little is known about the molecular mechanisms underlying this phenomenon. In this study, we conducted a gain-of-function screening of genes that confer cold resistance to cold-vulnerable human cells using a cDNA library constructed from the Syrian hamster, a mammalian hibernator, and identified Gpx4 as a potent suppressor of cold-induced cell death. Additionally, genetic deletion of or pharmacological inhibition of Gpx4 revealed that Gpx4 is necessary for suppressing lipid peroxidation specifically under cold in hamster cell lines. Genetic disruption of other ferroptosis-suppressing pathways, namely biopterin synthesis and mitochondrial or plasma membrane CoQ reduction pathways, also accelerated cold-induced cell death under Gpx4 dysfunction. Collectively, ferroptosis-suppressing pathways protect the cells of a mammalian hibernator from cold-induced cell death and the augmentation of these pathways renders cold resistance to cells of non-hibernators, including humans.

Abstract To describe the diagnosis and successful treatment of systemic francisellosis in a dog. An 11‐year‐old female spayed Labrador retriever presented for progressive lethargy, hyporexia, and cough. The dog was febrile with a neutrophilia, nonregenerative anemia, thrombocytopenia, and had increased activity in serum of liver‐derived enzymes. Francisella philomiragia was isolated from aerobic blood culture. The dog was treated for 6 weeks with enrofloxacin orally. Repeated aerobic blood cultures after 2 and 6 weeks of antibiotic therapy were negative. The dog was clinically normal 7 months after diagnosis with no evidence of relapse.

Language proficiency policies act as critical gatekeepers for multilingual international students (MIS) who intend to pursue an education at English-medium universities. The current study revisits the question of the predictive validity of a standardized English proficiency exam—International English Language Testing System (IELTS)—in comparison with 10 indices of written syntactic complexity. This study addressed the following questions: (a) Do MIS’ overall proficiency and writing subtest scores (IELTS) predict their grade point averages (GPAs) in graduate school? (b) Which syntactic complexity indices in online written production (subordination, coordination, length of production, and degree of phrasal sophistication) predict MIS’ GPAs in graduate school? and (c) Which syntactic complexity indices remain predictive of their GPAs when participants’ English proficiency scores (overall and/or writing IELTS scores) are added into the model? Participants’ IELTS scores, GPAs, and asynchronous discussion postings (224) were analyzed through three linear regressions models. Results confirmed that while IELTS composite and writing subscores do predict academic success of graduate MIS, writing scores are better predictors. After accounting for the contributions of overall and writing subscores, mean length of sentence and T-unit remained significant predictors of academic outcomes, explaining a considerable amount of variance in GPAs, beyond that of IELTS. Equity implications for the admissions of MIS are considered.

Study region: Mimoso Alluvial Valley, Semiarid Brazil. Study focus: This study aims to assess the influence of climate variability and exploitation on groundwater accessibility, quantity and quality, based on a spatiotemporal data analysis from long-term monitoring field campaigns, conducted monthly from 2000 to 2019. This study successfully identified representative stable monitoring points, piezometers and wells, for piezometric and salinity levels employing the technique of relative differences, taking into account aquifer hydraulic properties. Trend analysis was then carried out adopting the Mann-Kendall Method, Sen's Slope test, Pettitt test, and the Seasonal Trend decomposition through the Loess (STL) method. Principal Component Multivariate Analysis (PCA) was also employed to validate long-term analysis of annual groundwater levels, rainfall, evapotranspiration, and pumping rates. New hydrological insights for the region: The spatial-temporal variability of salinity is closely linked to the salt balance in regions with low hydraulic permeability and groundwater use for irrigation. Groundwater salinity presented a strong link between rainfall and groundwater levels. A temporal rise in evapotranspiration alongside declines in rainfall and groundwater levels was observed, potentially exacerbating groundwater scarcity alongside intensified aquifer exploitation. Furthermore, groundwater salinity presented a decreasing trend for the pumping wells and no trends for piezometers. This could potentially disrupt future irrigation plans and cause long-term water shortages in a region already under severe water scarcity. Hence, groundwater pumping constitutes a positive management alternative for controlling groundwater salinity in the domain.

Abstract Bontebok (Damaliscus pygargus pygargus) and blesbok (D. p. phillipsi) are classified as separate sub‐species. The blesbok has a widespread distribution throughout South Africa and is listed as least concern by the International Union for Conservation of Nature (IUCN) Red List of Threatened Species. Bontebok on the other hand is endemic within the Cape Floristic Region of the Western Cape in South Africa and has been listed as near‐threatened species on the IUCN Red List of Threatened Species. Bontebok populations experienced a severe bottleneck and were brought back from the brink of extinction in the 1830s. Currently, the subspecies is threatened by hybridisation with blesbok resulting in fertile offspring. To date, molecular investigations using neutral markers have determined that genetic diversity in pure South African bontebok was significantly lower than in pure blesbok. Here, we investigated genetic diversity in bontebok, blesbok and hybrid individuals using microsatellites and an adaptive marker (toll‐like receptor two (TLR2)). The study of single nucleotide polymorphisms (SNPs) revealed five mutations in TLR2 in different individuals and subspecies of D. pygargus. This included three non‐synonymous and two synonymous mutations. The three amino acid substitution mutations were predicted to have no effect on protein function. Two of the five mutations, one of which resulted in an amino acid substitution, were not present in bontebok. The other three mutations were present to varying frequencies in the three groups. We confirm low adaptive and neutral diversity in bontebok. These mutations provide insights into the genetic diversity and relationships among the two sub‐species of D. pygargus and may have implications for their conservation and management.

Ticks are important ectoparasites of domestic animals, wild animals and humans. They spread a variety of infective agents such as protozoans, viruses, and bacteria. Cattle reared by smallholder farmers are susceptible to ticks and tick-borne pathogens due to the type of production system practiced by the farmers. Hence, this review was focused on the occurrence of ticks and tick-borne pathogens in cattle reared by smallholder farmers in South Africa. The systematic search produced a total of 13,408 articles from four databases, and after screening processes, the review utilized 23 articles published between 1983 and 2023. A total of 26 tick species belonging to seven genera were identified in the reviewed articles, with Rhipicephalus (Boophilus) decoloratus and Rhipicephalus evertsi evertsi being the most frequently reported tick species in South Africa followed by Amblyomma hebreum, Rhipicephalus appendiculatus, Hyalomma marginatum rufipes, Rhipicephalus microplus, Rhipicephalus follis, Rhipicephalus gertrudae and Hyalomma truncatum. The most frequently reported tick-borne pathogens across the provinces included Babesia bigemina, Babesia bovis, and Anaplasma marginale, with Eastern Cape Province accounting for most of the records followed by KwaZulu-Natal and Mpumalanga Provinces. The findings of this review confirm that cattle reared by smallholder farmers harbour various ticks and tick-borne pathogens of veterinary, public health and economic importance, and regular monitoring of tick infestations in South Africa is recommended to avoid disease outbreaks.

Summary: SARS-CoV-2 infection in children typically results in asymptomatic or mild disease. There is a paucity of studies on SARS-CoV-2 antiviral immunity in African children. We investigated SARS-CoV-2-specific T cell responses in 71 unvaccinated asymptomatic South African children who were seropositive or seronegative for SARS-CoV-2. SARS-CoV-2-specific CD4+ T cell responses were detectable in 83% of seropositive and 60% of seronegative children. Although the magnitude of the CD4+ T cell response did not differ significantly between the two groups, their functional profiles were distinct, with SARS-CoV-2 seropositive children exhibiting a higher proportion of polyfunctional T cells compared to their seronegative counterparts. The frequency of SARS-CoV-2-specific CD4+ T cells in seronegative children was associated with the endemic human coronavirus (HCoV) HKU1 IgG response. Overall, the presence of SARS-CoV-2-responding T cells in seronegative children may result from cross-reactivity to endemic coronaviruses and could contribute to the relative protection from disease observed in SARS-CoV-2-infected children.

In 2024, the average annual premium for employer-sponsored family health coverage was $25,572, an increase of $1,604 (7 percent) from 2023. Over the course of the past five years, the average family premium has increased 24 percent, which is similar to growth seen in inflation (23 percent) and wages (28 percent). On average, covered workers contributed 16 percent ($1,368) of the cost of single coverage and 25 percent ($6,296) of the cost of family coverage. The average general annual deductible for single coverage for workers with a deductible was $1,787, similar to that in recent years but 47 percent higher than a decade ago. In 2024, 18 percent of large firms offering health benefits, including 28 percent of those with 5,000 or more employees, covered GLP-1 antagonists for weight loss. Large employers were more likely to perceive their overall provider networks as broader than their networks for mental health and substance use conditions.

Sickle cell disease (SCD) and systemic lupus erythematosus (SLE) are two uncommon disorders each characterized by multisystemic manifestations. Individuals with SCD exhibit abnormalities in the complement pathway, which may predispose patients to develop autoimmune disorders such as SLE. As many manifestations of SLE mimic those of SCD, diagnosis and therapeutic management of SLE in a patient with known SCD may be delayed. In this study, we describe our institutional experience of diagnosing and managing concomitant SCD and SLE. We offer insights into the complex interplay between these conditions to enhance early recognition and effective management of concurrent SCD and SLE., (© 2024 Wiley Periodicals LLC.)

The impact of environmental enrichment (EE) on natural rewards, including social and appetitive rewards, was investigated in male Swiss mice. EE, known for providing animals with various stimuli, was assessed for its effects on conditioned place preference (CPP) associated with ethanol and social stimuli. We previously demonstrated that EE increased the levels of the prosocial neuropeptide oxytocin (OT) in the hypothalamus and enhanced ethanol rewarding effects via an oxytocinergic mechanism. This study also investigated the impact of EE on social dominance and motivation for rewards, measured OT-mediated phospholipase C (PLC) activity in striatal membranes, and assessed OT expression in the hypothalamus. The role of dopamine in motivating rewards was considered, along with the interaction between OT and D 1 receptors (DR) in the nucleus accumbens (NAc). Results showed that EE mice exhibited a preference for ethanol reward over social reward, a pattern replicated by the OT analogue Carbetocin. EE mice demonstrated increased social dominance and reduced motivation for appetitive taste stimuli. Higher OT mRNA levels in the hypothalamus were followed by diminished OT receptor (OTR) signaling activity in the striatum of EE mice. Additionally, EE mice displayed elevated D 1 R expression, which was attenuated by the OTR antagonist (L-368-889). The findings underscore the reinforcing effect of EE on ethanol and social rewards through an oxytocinergic mechanism. Nonetheless, they suggest that mechanisms other than the prosocial effect of EE may contribute to the ethanol pro-rewarding effect of EE and Carbetocin. They also point towards an OT-dopamine interaction potentially underlying some of these effects., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Competing Interests: KF is President of the Faculty of Public Health. GF is President of the UK Association of Directors of Public Health, a member of the gambling related harm strategic steering group for Yorkshire and the Humber, which is funded by a regulatory settlement awarded by the UK Gambling Commission, and a member of the UK National Screening Committee. WR is Chief Executive of the Royal Society for Public Health and an honorary member of the Faculty of Public Health. MR is President of the Public Health and Epidemiology Section, Royal Society of Medicine, and immediate Past-President of the Faculty of Public Health. We declare no other competing interests.

The mechanical properties of the mammalian cell regulate many cellular functions and are largely dictated by the cytoskeleton, a composite network of protein filaments, including actin, microtubules, and intermediate filaments. Interactions between these distinct filaments give rise to emergent mechanical properties that are difficult to generate synthetically, and recent studies have made great strides in advancing our understanding of the mechanical interplay between actin and microtubule filaments. While intermediate filaments play critical roles in the stress response of cells, their effect on the rheological properties of the composite cytoskeleton remains poorly understood. Here, we use optical tweezers microrheology to measure the linear viscoelastic properties and nonlinear stress response of composites of actin and vimentin with varying molar ratios of actin to vimentin. We reveal a surprising, nearly opposite effect of actin-vimentin network mechanics compared to single-component networks in the linear versus nonlinear regimes. Namely, the linear elastic plateau modulus and zero-shear viscosity are markedly reduced in composites compared to single-component networks of actin or vimentin, whereas the initial response force and stiffness are maximized in composites versus single-component networks in the nonlinear regime. While these emergent trends are indicative of distinct interactions between actin and vimentin, nonlinear stiffening and longtime stress response appear to both be dictated primarily by actin, at odds with previous bulk rheology studies. We demonstrate that these complex, scale-dependent effects arise from the varied contributions of network density, filament stiffness, non-specific interactions, and poroelasticity to the mechanical response at different spatiotemporal scales. Cells may harness this complex behavior to facilitate distinct stress responses at different scales and in response to different stimuli to allow for their hallmark multifunctionality.