Degradation of ice-wedge polygons leads to increased fluxes of water and DOC.

Ice-wedge polygon landscapes make up a substantial part of high-latitude permafrost landscapes. The hydrological conditions shape how these landscapes store and release organic carbon. However, their coupled water­carbon dynamics are poorly understood as field measurements are sparse in smaller catchments and coupled hydrology-dissolved organic carbon (DOC) models are not tailored for these landscapes. Here we present a model that simulates the hydrology and associated DOC export of high-centered and low-centered ice-wedge polygons and apply the model to a small catchment with abundant polygon coverage along the Yukon Coast, Canada. The modeled seasonal pattern of water and carbon fluxes aligns with sparse field data. These modeled seasonal patterns indicate that early-season runoff is mostly surficial and generated by low-centered polygons and snow trapped in troughs of high-centered polygons. High-centered polygons show potential for deeper subsurface flow under future climate conditions. This suggests that high-centered polygons will be responsible for an increasing proportion of annual DOC export compared to low-centered polygons. Warming likely shifts low-centered polygons to high-centered polygons, and our model shows that this shift will cause a deepening of the active layer and a lengthening of the thawing season. This, in turn, intensifies seasonal runoff and DOC flux, mainly through its duration. Our model provides a physical hypothesis that can be used to further quantify and refine our understanding of hydrology and DOC export of arctic ice-wedge polygon terrain.

Effect of Climatic Condition, Type of Trough and Water Cleanliness on Drinking Behavior in Dairy Cows.

Increasing ambient temperatures lead to higher water intake and higher risks of microbial growth in cattle troughs. This study aims to analyze drinking water quality and dairy cows' drinking behavior (n = 8081 drinking episodes) on a commercial farm with 135 and 144 lactating cows in two climatic conditions, considering trough type and cleanliness, respectively. Daily video recording was conducted at two trough types (two open troughs, 70 L; two-valve troughs, variable volume of 5-15 L) in the first two hours after feeding (n = 60 days in total) under cold (December 2019-February 2020) and warm ambient temperatures (September 2021). The trough cleaning scheme allowed cows to access either cleaned or uncleaned troughs in each system. Water quality was tested daily and analyzed at the beginning and end of the trials. In warmer ambient temperatures, fewer and-at uncleaned troughs and open troughs-shorter drinking episodes were recorded, with longer but fewer water intake periods, longer drinking breaks, and fewer sips (p < 0.0001). Considering the drinking episodes, respectively, water intake and drinking breaks in number and duration, the number of sips and the number of agonistic behaviors might optimize dairy cow water supply and hygiene management.

Translation Fidelity and Respiration Deficits in CLPP-Deficient Tissues: Mechanistic Insights from Mitochondrial Complexome Profiling.

The mitochondrial matrix peptidase CLPP is crucial during cell stress. Its loss causes Perrault syndrome type 3 (PRLTS3) with infertility, neurodegeneration, and a growth deficit. Its target proteins are disaggregated by CLPX, which also regulates heme biosynthesis via unfolding ALAS enzymes, providing access for pyridoxal-5'-phosphate (PLP). Despite efforts in diverse organisms with multiple techniques, CLPXP substrates remain controversial. Here, avoiding recombinant overexpression, we employed complexomics in mitochondria from three mouse tissues to identify endogenous targets. A CLPP absence caused the accumulation and dispersion of CLPX-VWA8 as AAA+ unfoldases, and of PLPBP. Similar changes and CLPX-VWA8 co-migration were evident for mitoribosomal central protuberance clusters, translation factors like GFM1-HARS2, the RNA granule components LRPPRC-SLIRP, and enzymes OAT-ALDH18A1. Mitochondrially translated proteins in testes showed reductions to <30% for MTCO1-3, the mis-assembly of the complex IV supercomplex, and accumulated metal-binding assembly factors COX15-SFXN4. Indeed, heavy metal levels were increased for iron, molybdenum, cobalt, and manganese. RT-qPCR showed compensatory downregulation only for Clpx mRNA; most accumulated proteins appeared transcriptionally upregulated. Immunoblots validated VWA8, MRPL38, MRPL18, GFM1, and OAT accumulation. Co-immunoprecipitation confirmed CLPX binding to MRPL38, GFM1, and OAT, so excess CLPX and PLP may affect their activity. Our data mechanistically elucidate the mitochondrial translation fidelity deficits which underlie progressive hearing impairment in PRLTS3.

Ultra-Deep Sequencing Reveals the Mutational Landscape of Classical Hodgkin Lymphoma.

The malignant Hodgkin and Reed Sternberg (HRS) cells of classical Hodgkin lymphoma (cHL) are scarce in affected lymph nodes, creating a challenge to detect driver somatic mutations. As an alternative to cell purification techniques, we hypothesized that ultra-deep exome sequencing would allow genomic study of HRS cells, thereby streamlining analysis and avoiding technical pitfalls. To test this, 31 cHL tumor/normal pairs were exome sequenced to approximately 1,000× median depth of coverage. An orthogonal error-corrected sequencing approach verified >95% of the discovered mutations. We identified mutations in genes novel to cHL including: CDH5 and PCDH7, novel stop gain mutations in IL4R, and a novel pattern of recurrent mutations in pathways regulating Hippo signaling. As a further application of our exome sequencing, we attempted to identify expressed somatic single-nucleotide variants (SNV) in single-nuclei RNA sequencing (snRNA-seq) data generated from a patient in our cohort. Our snRNA analysis identified a clear cluster of cells containing a somatic SNV identified in our deep exome data. This cluster has differentially expressed genes that are consistent with genes known to be dysregulated in HRS cells (e.g., PIM1 and PIM3). The cluster also contains cells with an expanded B-cell clonotype further supporting a malignant phenotype. This study provides proof-of-principle that ultra-deep exome sequencing can be utilized to identify recurrent mutations in HRS cells and demonstrates the feasibility of snRNA-seq in the context of cHL. These studies provide the foundation for the further analysis of genomic variants in large cohorts of patients with cHL. SIGNIFICANCE: Our data demonstrate the utility of ultra-deep exome sequencing in uncovering somatic variants in Hodgkin lymphoma, creating new opportunities to define the genes that are recurrently mutated in this disease. We also show for the first time the successful application of snRNA-seq in Hodgkin lymphoma and describe the expression profile of a putative cluster of HRS cells in a single patient.

Effect of Rumen-Protected Methionine on Metabolic Profile of Liver, Muscle and Blood Serum Samples of Growing German Simmental Bulls Fed Protein-Reduced Diets.

This study aimed to determine the metabolic response of growing German Simmental bulls fed rations low in crude protein (CP) supplemented with rumen-protected methionine (RPMET). In total, 69 bulls (on average 238 ± 11 days of age at start and 367 ± 25 kg of bodyweight) were assigned to three dietary treatments (n = 23/group): Positive control (CON; 13.7% CP; 2.11 g methionine/kg DM), negative control deficient in CP (RED; 9.04% CP; 1.56 g methionine/kg DM) and crude protein-deficient ration supplemented with RPMET (RED+RPMET; 9.04% CP; 2.54 g methionine/kg DM). At slaughter, samples of liver, muscle and blood serum were taken and underwent subsequent metabolomics profiling using a UHPLC-QTOF-MS system. A total of 6540 features could be detected. Twenty metabolites in the liver, five metabolites in muscle and thirty metabolites in blood serum were affected (p < 0.05) due to dietary treatments. In total, six metabolites could be reliably annotated and were thus subjected to subsequent univariate analysis. Reduction in dietary CP had minimal effect on metabolite abundance in target tissues of both RED and RED+RPMET bulls as compared to CON bulls. The addition of RPMET altered the hepatic anti-oxidant status in RED+RPMET bulls compared to both RED and CON bulls. Results exemplify nutrient partitioning in growing German Simmental bulls: bulls set maintenance as the prevailing metabolic priority (homeostasis) and nutrient trafficking as the second priority, which was directed toward special metabolic functions, such as anti-oxidant pathways.

DNA Methylation Signatures of Functional Somatic Syndromes: Systematic Review.

OBJECTIVE: Functional somatic syndromes (FSS) are highly prevalent across all levels of health care. The fact that they are characterized by medically unexplained symptoms, such as fatigue and pain, raises the important question of their underlying pathophysiology. Psychosocial stress represents a significant factor in the development of FSS and can induce long-term modifications at the epigenetic level. The aim of this review was to systematically review, for the first time, whether individuals with FSS are characterized by specific alterations in DNA methylation. METHODS: MEDLINE and PsycINFO were searched from the first available date to September 2022. The inclusion criteria were as follows: a) adults fulfilling the research diagnostic criteria for chronic fatigue syndrome, fibromyalgia syndrome, and/or irritable bowel syndrome; b) healthy control group; and c) candidate-gene or genome-wide study of DNA methylation. RESULTS: Sixteen studies ( N = 957) were included. In candidate-gene studies, specific sites within NR3C1 were identified, which were hypomethylated in individuals with chronic fatigue syndrome compared with healthy controls. In genome-wide studies in chronic fatigue syndrome, a hypomethylated site located to LY86 and hypermethylated sites within HLA-DQB1 were found. In genome-wide studies in fibromyalgia syndrome, differential methylation in sites related to HDAC4 , TMEM44 , KCNQ1 , SLC17A9 , PRKG1 , ALPK3 , TFAP2A , and LY6G5C was found. CONCLUSIONS: Individuals with chronic fatigue syndrome and fibromyalgia syndrome seem to be characterized by altered DNA methylation of genes regulating cellular signaling and immune functioning. In chronic fatigue syndrome, there is preliminary evidence for these to be implicated in key pathophysiological alterations, such as hypocortisolism and low-grade inflammation, and to contribute to the debilitating symptoms these individuals experience. PREREGISTRATION: PROSPERO identifier: CRD42022364720.

Learning to Fly.

In this narrative medicine essay, an infectious diseases fellow understands while grieving her father that care cannot always be measured in cure but rather in listening and loving.

T-BET and EOMES sustain mature human NK cell identity and antitumor function.

Since the T-box transcription factors (TFs) T-BET and EOMES are necessary for initiation of NK cell development, their ongoing requirement for mature NK cell homeostasis, function, and molecular programming remains unclear. To address this, T-BET and EOMES were deleted in unexpanded primary human NK cells using CRISPR/Cas9. Deleting these TFs compromised in vivo antitumor response of human NK cells. Mechanistically, T-BET and EOMES were required for normal NK cell proliferation and persistence in vivo. NK cells lacking T-BET and EOMES also exhibited defective responses to cytokine stimulation. Single-cell RNA-Seq revealed a specific T-box transcriptional program in human NK cells, which was rapidly lost following T-BET and EOMES deletion. Further, T-BET- and EOMES-deleted CD56bright NK cells acquired an innate lymphoid cell precursor-like (ILCP-like) profile with increased expression of the ILC-3-associated TFs RORC and AHR, revealing a role for T-box TFs in maintaining mature NK cell phenotypes and an unexpected role of suppressing alternative ILC lineages. Our study reveals the critical importance of sustained EOMES and T-BET expression to orchestrate mature NK cell function and identity.

Perspectives of individuals on reducing meat consumption to mitigate climate change: protocol for a scoping review.

INTRODUCTION: High consumption of red and processed meat increases the risk of several chronic diseases. Many people, especially in high-income countries, eat more meat than recommended by nutritional and health agencies. Meat production also has negative impacts on the environment and contributes to climate change. Therefore, climate protection, besides health or animal welfare, could motivate individuals to eat less meat. Willingness to reduce meat consumption and motives to do so are not yet fully understood. METHODS AND ANALYSIS: Based on the Preferred Reporting Items for Systematic reviews and Meta-Analyses extension for Scoping Reviews (PRISMA-ScR) extended guidelines, a scoping review of peer-reviewed original studies will be conducted to address three questions: What is the evidence regarding (1) the willingness of individuals to reduce meat consumption to mitigate climate change, (2) the awareness of individuals about the link between their meat consumption and the potential to mitigate climate change and (3) individuals having reduced meat consumption for the reason of climate protection? We will search the databases Medline (via PubMed), Scopus, Embase, Greenfile (via Ebsco) and PsynDex/CurrentContent/Agris (via Livivo) using a systematic search string. Studies from 2015 onwards, published in English, German, Danish or Dutch, will be included. We will include observational studies, qualitative studies, intervention studies (if they include surveys) and reviews. Data will be summarised in a narrative synthesis, comprising methods, population characteristics, meat type under study, indicators measured and limitations. Key findings will be grouped according to the research questions. This scoping review will help clarify the role of climate protection in individual reduction of meat consumption and identify research gaps in this field. ETHICS AND DISSEMINATION: Formal ethical approval is not required, as primary data will not be collected in this study. Findings of this scoping review will be presented at scientific conferences and published in a peer-reviewed journal. TRIAL REGISTRATION NUMBER: https://doi.org/10.17605/OSF.IO/MWB85.

Both radiographical and pathological lymph node statuses are independent predictors for survival following neoadjuvant chemotherapy and radical cystectomy for cT3/4 or cN+ bladder cancer.

INTRODUCTION: Urothelial bladder cancer (UBC) with clinical suspicion of locally advanced growth or pelvic lymphogenic spread has a high risk of progression and death. PATIENTS AND METHODS: Bladder cancer patients with locally advanced (cT3/4) tumor growth or suspected pelvic lymphogenic spread (cN+) were treated with preoperative cisplatin-containing chemotherapy and consolidative cystectomy with pelvic lymphadenectomy. We aimed to identify prognostic factors and describe the patients' oncological outcome. RESULTS: A complete dataset including follow-up data was available for 96 patients. In a univariate analysis, we identified cN stage (cN+ vs cN-, HR 2.7, 95% CI 1.3-6.0), response to chemotherapy (HR 0.2, 95% CI 0.1-0.5), ypT stage (ypT0/is/1 vs ypT2-4, HR 3.1, 95% CI 1.4-6.8), ypN stage (ypN + vs ypN-, HR 7.9, 95% CI 3.7-17.0), resection status (HR 4.4, 95% CI HR 1.5-13.0) as significantly associated with cancer-specific survival. In a multivariate regression analysis, both cN and ypN statuses were validated as independent prognostic factors for cancer-specific survival (cN: HR 2.6, 95% CI 1.1-6.1; ypN: HR 5.5, 95% CI 2.0-15.1). DISCUSSION: Lymph node status was identified as a prognostic marker in a high-risk cohort of UBC patients treated with inductive chemotherapy and cystectomy. Establishing cN status as a prognosticator underlines the necessity to aggressively treat these patients despite reported impreciseness of imaging procedures in UCB. Patients with histologically positive lymph nodes following preoperative chemotherapy have a very poor prognosis, and thus, the need for adjuvant systemic treatment is emphasized. CONCLUSION: Both clinically and pathologically affected lymph nodes convey a poor prognosis in bladder cancer and necessitate aggressive treatment.

Cause and Effect Analysis between Influencing Factors Related to Environmental Conditions, Hunting and Handling Practices and the Initial Microbial Load of Game Carcasses.

Environmental, hunting and handling factors affect the microbial load of hunted game and the resulting meat products. The aim of this study was to systematically investigate the influence of several factors on the initial microbial load (IML) of game carcasses during the early hunting chain. Eviscerated roe deer body cavities (n = 24) were investigated in terms of total viable count and the levels of Pseudomonas spp., Lactobacillus spp., Enterobacteriaceae and Escherichia coli (E. coli). Furthermore, a risk analysis based on the obtained original IML data, literature search and a Failure Mode and Effects Analysis (FMEA) was performed. The IML could be explained in a regression model by factors including the higher body weight (BW), damaged gastrointestinal tract by the shot, ambient temperature or rain. The levels of Lactobacillus spp. (p = 0.0472), Enterobacteriaceae (p = 0.0070) and E. coli (p = 0.0015) were lower on the belly flap surface when gloves were used during evisceration. The literature search revealed that studies examining influencing factors (IF) on the IML of game carcasses found contradictory effects of the comparable IF on IML. Potential handling failures may lead to a higher IML of game carcasses during the early hunting chain ranked by FMEA. Several handling practices for game carcasses are recommended, such as ensuring efficient cooling of heavier BW carcasses to limit bacterial growth or eviscerating heavier carcasses before lighter ones.

PEGylated Gold Nanoparticles Target Age-Associated B Cells In Vivo.

Engineered gold nanoparticles (GNPs) have become a useful tool in various therapeutic and diagnostic applications. Uncertainty remains regarding the possible impact of GNPs on the immune system. In this regard, we investigated the interactions of polymer-coated GNPs with B cells and their functions in mice. Surprisingly, we observed that polymer-coated GNPs mainly interact with the recently identified subpopulation of B lymphocytes named age-associated B cells (ABCs). Importantly, we also showed that GNPs did not affect cell viability or the percentages of other B cell populations in different organs. Furthermore, GNPs did not activate B cell innate-like immune responses in any of the tested conditions, nor did they impair adaptive B cell responses in immunized mice. Together, these data provide an important contribution to the otherwise limited knowledge about GNP interference with B cell immune function, and demonstrate that GNPs represent a safe tool to target ABCs in vivo for potential clinical applications.

Evaluation of the Occurrence of Staphylococcaceae with Reduced Susceptibility to Cefoxitin in Wild Ungulates in Brandenburg, Germany, Based on Land Use-Related Factors.

Interactions between natural and human-used environments have a significant influence on the spread of antimicrobial resistance in wild ecosystems. Despite current knowledge, fundamental questions about the degree of impact of land use-related factors on the spread of antimicrobial-resistant staphylococci in European wild game animal populations have not yet been answered with certainty. In this study, we evaluated the occurrence of Staphylococcaceae showing reduced susceptibility to cefoxitin in nasal swabs of fallow deer (Dama dama), red deer (Cervus elaphus), roe deer (Capreolus capreolus), and wild boar (Sus scrofa) hunted in Brandenburg, Germany. Evaluations were focused on the use of open-source data regarding the extent as well as the degree of land use, especially for settlement or animal husbandry. Results showed that the detection rate of Staphylococcaceae showing a non-wild-type phenotype for cefoxitin differed between animal species of the studied hunting districts. Statistical analyses of results combined with data on land use features revealed that a high density of cattle or poultry in a county may be associated with an increased detection rate in roe deer or wild boar, respectively. Furthermore, positive correlations were determined between the prevalence of non-wild-type Staphylococcaceae in roe deer or fallow deer and the proportional extent of surface water bodies in the corresponding area. The presented approach establishes a general basis for a risk-oriented assessment of the effects of human activities on the epidemiology of transmissible microorganisms in the human-animal-environment interface, including antimicrobial-resistant bacteria. IMPORTANCE Intensive research regarding the impact of land use-related factors on the prevalence and distribution of antimicrobial-resistant Staphylococcaceae in game ungulate populations is necessary for adequately determining risks related to interactions between wild animals, domestic animals, and humans in common geographic locations. This systematic approach for the analysis of the observations in specific hunting districts of Brandenburg, Germany, adds an innovative value to the research strategy of antimicrobial resistance in wild game animals, which is in accordance with current recommendations worldwide. Thus, results and information obtained in this study build a relevant foundation for future risk assessment regarding the safety of game products. Furthermore, the data generated represent an important basis for improving existing guidelines in land use practices and hunting practices. The use of existing open source data collections provided by official governmental and nongovernmental entities increases not only the impact but also the applicability and comparability of information beyond the regional level.

Glioblastoma hijacks neuronal mechanisms for brain invasion.

Glioblastomas are incurable tumors infiltrating the brain. A subpopulation of glioblastoma cells forms a functional and therapy-resistant tumor cell network interconnected by tumor microtubes (TMs). Other subpopulations appear unconnected, and their biological role remains unclear. Here, we demonstrate that whole-brain colonization is fueled by glioblastoma cells that lack connections with other tumor cells and astrocytes yet receive synaptic input from neurons. This subpopulation corresponds to neuronal and neural-progenitor-like tumor cell states, as defined by single-cell transcriptomics, both in mouse models and in the human disease. Tumor cell invasion resembled neuronal migration mechanisms and adopted a Lévy-like movement pattern of probing the environment. Neuronal activity induced complex calcium signals in glioblastoma cells followed by the de novo formation of TMs and increased invasion speed. Collectively, superimposing molecular and functional single-cell data revealed that neuronal mechanisms govern glioblastoma cell invasion on multiple levels. This explains how glioblastoma's dissemination and cellular heterogeneity are closely interlinked.

FOCUS on women: Program evaluation of a pilot probation and primary care transitions clinic collaboration☆.

Gender-specific probation programs are needed for rising numbers of women on probation, as women's paths to probation differ from men's and are not as frequently addressed. Substance use, mental illness, health disparities, and barriers to treatment can contribute to women clients' unsuccessful completion of probation. The current study describes a process evaluation of the pilot Female Offenders Can Ultimately Succeed (FOCUS) program, which aimed to meet women probation clients' complex needs and improve their probation outcomes. Two women FOCUS Probation Officers had smaller-than-typical caseloads of 132 Medium- to Greatest-Risk women clients and received ongoing training in motivational and trauma-specific supervision strategies. To facilitate linkage to all needed treatment and social services, clients were referred to a primary care Transitions Clinic. Comparing non-FOCUS to FOCUS women clients with Violations of Probation, FOCUS clients had more successful completions of probation (higher non-punitive Restorations of Probation and lower punitive Revocations of Probation). At the Transitions Clinic, 52% of FOCUS clients received treatment. FOCUS clients and stakeholders provided narrative feedback reflecting a range of experiences in FOCUS, guiding future program implementation efforts. Promising outcomes from FOCUS demonstrate the potential of theory-based supervision and probation-medical collaboration to facilitate women clients' success in probation.

Standardised Sampling Approach for Investigating Pathogens or Environmental Chemicals in Wild Game at Community Hunts.

Wildlife may host pathogens and chemicals of veterinary and public health relevance, as well as pathogens with significant economic relevance for domestic livestock. In conducting research on the occurrence and distribution of these agents in wildlife, a major challenge is the acquisition of a sufficient number of samples coupled with efficient use of manpower and time. The aim of this article is to present the methodology and output of a sampling approach for game animals, which was implemented from 2017/18 to 2020/21 at drive hunts in Brandenburg, Germany. The central element was a framework agreement with the BImA, whereby federal forest officials and other hunters collected most of the samples during field dressing. Further samples of game carcasses were obtained by scientists during subsequent gathering at a collection point. Altogether, 3185 samples from 938 wild ungulates of four species were obtained for various studies analysing-in this case-food-borne agents in game animals. Sampling was representative and reflected the proportional distribution of ungulate species hunted in Brandenburg. Hunting district and hunting season strongly influenced hunting bag and hence sampling success. This sampling approach was demonstrated to be a suitable basis for monitoring programs, that can be adapted to other regions.

A risk-oriented evaluation of biofilm and other influencing factors on biological quality of drinking water for dairy cows.

Despite the importance of livestock drinking water quality on animal physiology, welfare, and performance, influences such as biofilm formation on trough surfaces on microbial water quality are rarely researched. The objective of this study was to assess the microbial quality of water offered to lactating dairy cows and identify risk factors for poor water quality. We further aimed to determine the impact of biofilm formation on water quality and evaluate rapid test systems to score the hygiene status of dairy troughs on the farm. A total of 105 troughs located on 24 typical Western German dairy farms were sampled. Samples of livestock drinking water and biofilm were analyzed for aerobic total viable count (TVC), coliform count (CC), Escherichia coli, methicillin-resistant Staphylococcus aureus (MRSA), and other bacteria resistant to 3rd generation cephalosporins (CRB). Surface protein- and adenosine triphosphate (ATP)-rapid tests were evaluated to detect biofilm formation. The influence of 22 selected fixed and variable trough characteristics on impaired livestock drinking water quality was evaluated by calculating odds ratios. The average TVC, CC, and E. coli counts were 4.4 ±â€…0.06 (mean ±â€…SD), 1.7 ±â€…0.1, and 0.6 ±â€…0.1 log10 cfu per mL, respectively. CC was detectable in 94.3% of all water samples and E. coli in 48.6%. MRSA was found in pooled livestock drinking water samples of a single farm and CRB on three farms, suggesting that troughs might function as a reservoir of antibiotic-resistant bacteria, thereby contributing to an exchange of antibiotic-resistant bacteria between animals. Risk factors for the impairment of at least one microbial quality criteria (TVC, CC, or E. coli) increased significantly (P < 0.05) when using high-volume troughs, other trough materials than stainless steel, a lower distance to the milking parlor, heavy visible soiling, biofilm formation, and high ambient and high water temperatures. CC (r = 0.46; P < 0.001) and E. coli (r = 0.31; P < 0.01) of water samples correlated with their equivalent in biofilm and with the results of rapid tests on trough surfaces (0.31 > r > 0.19; P < 0.05). Addressing the identified risk factors could be an approach to ensure sufficient biological quality of livestock drinking water.

Although water is recognized as one of the most important nutrients for dairy cows, only limited research on its quality and potential risk factors for quality impairments is available. This study aimed to evaluate biological water quality on Western German dairy farms, identify potential risk factors for impairment, and evaluate rapid test systems to score the hygienic status of troughs. Water, biofilm, and trough surfaces of 105 water troughs on 24 dairy farms were sampled and analyzed. The bacterial load of sampled livestock drinking water was relatively high. Nearly half of all water samples were contaminated with Escherichia coli, and antibiotic-resistant bacteria were detectable on four farms. Identified risk factors for poor biological water quality that could be used to ensure sufficient water quality on-farm were: high-volume troughs, plastic or cast iron as trough material, a lower distance to the milking parlor, heavy visible soiling, biofilm formation, and high ambient and high water temperatures. Rapid test systems to evaluate trough surfaces correlated with biofilm and water analysis results and could be a useful tool for farmers to check water quality on-farm.

A novel fusion protein scaffold 18/12/TxM activates the IL-12, IL-15, and IL-18 receptors to induce human memory-like natural killer cells.

Natural killer (NK) cells are cytotoxic innate lymphoid cells that are emerging as a cellular immunotherapy for various malignancies. NK cells are particularly dependent on interleukin (IL)-15 for their survival, proliferation, and cytotoxic function. NK cells differentiate into memory-like cells with enhanced effector function after a brief activation with IL-12, IL-15, and IL-18. N-803 is an IL-15 superagonist composed of an IL-15 mutant (IL-15N72D) bound to the sushi domain of IL-15Rα fused to the Fc region of IgG1, which results in physiological trans-presentation of IL-15. Here, we describe the creation of a novel triple-cytokine fusion molecule, 18/12/TxM, using the N-803 scaffold fused to IL-18 via the IL-15N72D domain and linked to a heteromeric single-chain IL-12 p70 by the sushi domain of the IL-15Rα. This molecule displays trispecific cytokine activity through its binding and signaling through the individual cytokine receptors. Compared with activation with the individual cytokines, 18/12/TxM induces similar short-term activation and memory-like differentiation of NK cells on both the transcriptional and protein level and identical in vitro and in vivo anti-tumor activity. Thus, N-803 can be modified as a functional scaffold for the creation of cytokine immunotherapies with multiple receptor specificities to activate NK cells for adoptive cellular therapy.

Impact of Routine Management Procedures on the Welfare of Suckling Piglets.

Piglets often undergo several painful treatments during the initial days of their lives. In this review, we investigate the acute (i.e., immediate), short-, and long-term implications of piglet processing on behavioral, physiological, clinical, and performance parameters, and how welfare impairments depend on performance instead of sham procedure, alternative techniques, or the age of the piglets. Welfare indicators that have been used to determine the least distressing procedures and knowledge gaps with regard to the procedures are identified and discussed. Tail docking and especially piglet castration have been the most researched topics, whereas marking for identification has been rarely addressed. Few or no studies have investigated the effects of teeth resection and tail docking on piglets of different age groups. Additionally, results are often found to be inconsistent, highlighting the need for additional research to determine the optimal age for processing. Studies comparing different processing techniques have produced contradictory results, but ear notching, teeth clipping, hot cautery tail docking, and tearing during castration have been determined to result in increased pain. Generally, a shorter procedure duration can reduce stress, with operator training having a distinct impact on piglet welfare during processing. As such, these topics should be further investigated.