Regenerative anemia identification in cats: Red blood cell indices or morphology, what to use?

Background and Aim: Anemia, a clinical condition characterized by reduced erythrocytes, is often observed in cats. Regeneration indicates that the bone marrow can respond appropriately to anemia. The absolute reticulocyte count is the reference for differentiating regenerative and non-regenerative anemia, while red blood cell (RBC) indices and morphology provide supplementary information. This study aimed to identify anemia types and establish the most reliable RBC indices and morphology methods in agreement with the reference method. Materials and Methods: One hundred forty-five cases of cat anemia were prospectively classified using two methods: RBC indices and RBC morphology, and subsequently compared with the absolute reticulocyte count. Results: Based on RBC indices assessment, 27 cases (19%) exhibited regenerative anemia. Based on RBC morphology, 29 (20%) cases were identified as having regenerative anemia. Using the reticulocyte absolute count as a reference method, 34 (23.4%) cases of regenerative anemia were identified. The findings indicated that RBC indices and RBC morphology did not align in evaluating medullary regeneration and that there is a good degree of agreement between RBC morphology assessment and the reticulocyte absolute count in identifying regenerative anemias. Conclusion: Blood smear analysis of RBC morphology was more dependable for classifying regenerative anemia than RBC indices. Further studies should be conducted with a larger number of animals and that allow the identification of the cause of anemia and the monitoring of the animal.

Cognitive Dysfunction Screening in Peritoneal Dialysis Patients: A Cross-Sectional Study.

Background: Mild cognitive impairment (MCI) in peritoneal dialysis (PD) patients has been described as a risk factor for worse outcomes such as peritonitis, technique failure, and mortality. In this study, we aimed to determine the prevalence of MCI in a population of PD patients and identify the possible risk factors associated with MCI. Materials and Methods: We performed an observational, cross-sectional study to evaluate cognitive function using the Montreal Cognitive Assessment (MOCA) test and the Mini Mental State Examination (MMSE) test in PD patients. Patients with diagnosis of dementia or severe neurologic impairment, active cancer, or infection were excluded. Results: We evaluated 66 patients (mean age 60 years); 53% were male. Prevalence of MCI assessed by MOCA test and MMSE test was 65% and 33%, respectively. Predictors of MCI with MOCA test were higher age (P = 0.0001), lower education level (P = 0.005), need of a helper (P = 0.009), and continuous ambulatory PD modality (P = 0.019). Higher Charlson comorbidity index (P = 0.002), coronary artery disease (P = 0.006), and peripheral artery disease (P = 0.033) were also associated with MCI. Lower Kt/V (P = 0.012) and lower levels of normalized protein catabolic rate (nPCR; P < 0.000) were related to MCI. MCI patients had more episodes of peritonitis (P = 0.047). Multivariable analysis showed that lower education, Kt/V, and nPCR were the most relevant factors connected to MCI (P = 0.029, P = 0.037, and P = 0.019, respectively). Conclusion: In our PD population, MCI was detected in more than half of the patients. Patients with MCI were older, had lower education level, more disease burden, and higher risk for developing peritonitis. Lower Kt/V and nPCR levels were associated with MCI.

ILC2-derived LIF licences progress from tissue to systemic immunity.

Migration and homing of immune cells are critical for immune surveillance. Trafficking is mediated by combinations of adhesion and chemokine receptors that guide immune cells, in response to chemokine signals, to specific locations within tissues and the lymphatic system to support tissue-localized immune reactions and systemic immunity1,2. Here we show that disruption of leukaemia inhibitory factor (LIF) production from group 2 innate lymphoid cells (ILC2s) prevents immune cells leaving the lungs to migrate to the lymph nodes (LNs). In the absence of LIF, viral infection leads to plasmacytoid dendritic cells (pDCs) becoming retained in the lungs where they improve tissue-localized, antiviral immunity, whereas chronic pulmonary allergen challenge leads to marked immune cell accumulation and the formation of tertiary lymphoid structures in the lung. In both cases immune cells fail to migrate to the lymphatics, leading to highly compromised LN reactions. Mechanistically, ILC2-derived LIF induces the production of the chemokine CCL21 from lymphatic endothelial cells lining the pulmonary lymphatic vessels, thus licensing the homing of CCR7+ immune cells (including dendritic cells) to LNs. Consequently, ILC2-derived LIF dictates the egress of immune cells from the lungs to regulate tissue-localized versus systemic immunity and the balance between allergen and viral responsiveness in the lungs.

Coagulation Profile of the Healthy Miranda's Donkey.

Miranda's donkey, originating in northern Portugal, is an autochthonous breed that is deeply intertwined with the region's culture and economy. Knowledge of the physiological characteristics of the breed is important for its preservation, and several studies have been carried out, but none have focused on its coagulation profile. The aim of this study was to establish reference intervals (RIs) for coagulation in healthy Miranda's donkey and to assess the influence of sex and age. Blood samples from 75 clinically healthy animals were analyzed for seven coagulation parameters: four using IDEXX ProCyte Dx and three using Start® 4-Diagnostica-Stago. The RI values were calculated following the ASVCP guidelines and with the Reference Advisor V.2.1 software. To analyze the influence of sex and age, SPSS version 29 was used. No significant differences were found between sexes (p > 0.05), but statistically significant differences were found between ages (p < 0.05) for platelet count and plateletcrit (both higher in young animals). The RIs described here can help monitor health and guide the diagnosis and treatment of diseased Miranda's donkeys, contributing to their preservation. Our study encourages further research on coagulation in donkeys and the use of different methodologies to obtain information for veterinarians working with this species.

Mef2d potentiates type-2 immune responses and allergic lung inflammation.

Innate lymphoid cells (ILCs) and adaptive T lymphocytes promote tissue homeostasis and protective immune responses. Their production depends on the transcription factor GATA3, which is further elevated specifically in ILC2s and T helper 2 cells to drive type-2 immunity during tissue repair, allergic disorders, and anti-helminth immunity. The control of this crucial up-regulation is poorly understood. Using CRISPR screens in ILCs we identified previously unappreciated myocyte-specific enhancer factor 2d (Mef2d)-mediated regulation of GATA3-dependent type-2 lymphocyte differentiation. Mef2d-deletion from ILC2s and/or T cells specifically protected against an allergen lung challenge. Mef2d repressed Regnase-1 endonuclease expression to enhance IL-33 receptor production and IL-33 signaling and acted downstream of calcium-mediated signaling to translocate NFAT1 to the nucleus to promote type-2 cytokine-mediated immunity.

The Effect of Lemon Juice (Citrus limon L.) Treated with Melatonin on the Health Status and Treatment of K14HPV16 Mice.

Lemon is a fruit rich in antioxidant properties and has several health benefits, namely the reduction of skin edema and anticarcinogenic properties, which are due to its high content of bioactive compounds. Melatonin can improve and preserve the properties of lemon for longer and also has health benefits. The aim of this study was to evaluate the effects of oral administration of lemon juice after melatonin treatment on murinometric parameters of wild-type (WT) mice and transgenic mice carrying human papillomavirus (HPV). Two trials were performed for oral administration of the lemon extract compound: in drinking water and in diet. First of all, lemons were treated by immersion with melatonin at 10 mM. Then, lemons were squeezed, and the juice obtained was freeze-dried and stored to be subsequently added to drinking water or diet, according to the assay. Thus, mice were divided into eight groups in the drink assay (each with n = 5): group 1 (G1, WT, control), group 2 (G2, WT, 1 mL lemon), group 3 (G3, WT, 1.5 mL lemon), group 4 (G4, WT, 2 mL lemon), group 5 (G5, HPV16, control), group 6 (G6, HPV16, 1 mL lemon) group 7 (G6, HPV16, 1.5 mL lemon) and group 8 (G6, HPV16, 2 mL lemon). The diet assay was divided into four groups: group 1 (G1, WT, control), group 2 (G2, WT, 4 mL lemon), group 3 (G3, HPV16, control) and group 4 (G4, HPV16, 4 mL lemon). In the drink assay, the highest concentration of melatonin (308 ng/100 mL) was for groups 4 and 8, while in the food assay, there was only one concentration of melatonin (9.96 ng/g) for groups 2 and 4. Both trials lasted 30 days. During this time, body weight, food and water were recorded. Afterward, they were sacrificed, and samples were collected for different analyses. At the concentrations used, the lemon juice with melatonin had no adverse effects on the animals' health and showed a positive outcome in modifying weight gain and enhancing antioxidant activity in mice. Moreover, a reduction in the incidence of histological lesions was observed in treated animals. Further research is needed to better understand the effects of lemon extract on health and treatment outcomes in this animal model.

Serum Biochemistry Parameters of the Endangered Miranda's Donkey Breed: Reference Intervals and the Influence of Gender and Age.

Miranda's donkey is an endangered, autochthone breed from Northern Portugal. Understanding the physiological and pathological conditions of Miranda's donkey is crucial for the conservation of the breed. Our study aimed to establish reference intervals (RIs) for blood biochemistry parameters and to evaluate the influence of gender and age on these parameters. Blood samples from 75 clinically healthy animals were analyzed for 21 blood biochemistry parameters using Respons® 920 and Start® 4-Diagnostica-Stago. RIs were calculated according to the ASVCP guidelines, utilizing Reference Advisor software V. 2.1 and the statistical program SPSS version 29 to analyze the effects of gender and age. Significant gender-related differences (p < 0.05) were observed in cholesterol, chlorine, creatinine (CREA), glucose (GLU), sodium, and triglycerides (with higher values in females) and in aspartate aminotransferase, creatine phosphokinase (CK), gamma-glutamyl transferase, potassium, magnesium (Mg), and urea (with higher values in males). Age-related differences (p < 0.05) were noted for alkaline phosphatase, CK, fibrinogen, GLU, and phosphorus (higher in young animals) and for globulin, Mg, and total protein (higher in adults). The RIs described here are invaluable for assessing and monitoring the health status of individual animals and herds. Our study highlights the importance of considering gender and age in the interpretation of biochemical parameters, providing critical insights for the conservation and management of this endangered breed.

A Review on Blood Reference Values as a Valuable Marker of Wildlife Welfare in Erinaceus europaeus.

The western-European hedgehog (Erinaceus europaeus), in expanding its range towards human habitats, faces exposure to contaminants and biological agents, potentially leading to diseases associated with hematological and biochemical changes. As bioindicators of environmental pollution and carriers of zoonotic agents, hedgehogs play a crucial role in One Health studies, emphasizing the need for a comprehensive understanding of their clinical-pathological aspects. Exploring the blood reference values in healthy animals of this species is crucial for understanding and improving their well-being, and identifying possible diseases/pathogens that may affect its conservation and/or impact human health. This review is focused on analyzing the data available in the literature for Erinaceus europaeus blood reference intervals. A comprehensive literature review of the studies published in Europe is performed, highlighting their specificities, and emphasizing the need for continuous research in this field. Our final goal is to provide a crucial tool for assessing the health status of the species, and underscoring the significance of research in this specific domain.

Establishment of Reference Intervals of Hematological Parameters and Evaluation of Sex and Age Effect in the Miranda Donkey.

The Miranda donkey is an autochthonous Portuguese breed that is considered endangered. Several studies have been carried out on this breed, but to the authors' best knowledge, no studies have been conducted on their clinical pathology. The aims of this study were to determine the hematological reference intervals (RIs) in healthy Miranda donkeys and to estimate the influence of age and sex. Blood samples from 75 clinically healthy animals were analyzed for 22 hematological parameters on the IDEXX ProCyte Dx, an automated hematology analyzer previously validated for the species. The RIs were estimated following the ASVCP guidelines with the Reference Value Advisor software. Regarding sex, no significant differences were found between groups. Regarding age, significant statistical differences (p < 0.05) were observed for red blood cells, red cell distribution width, white blood cells, lymphocytes, monocytes, platelets, plateletcrit (higher mean in young animals), mean corpuscular volume, mean corpuscular haemoglobin, neutrophils and eosinophils (higher mean in adults). The RIs described here can be used to evaluate and monitor the health status of animals and herds, as well as to guide diagnoses or select fit and healthy animals for reproduction, contributing to the preservation of the breed.

Neuroprotective protein ADNP-dependent histone remodeling complex promotes T helper 2 immune cell differentiation.

Type 2 immune responses are critical in tissue homeostasis, anti-helminth immunity, and allergy. T helper 2 (Th2) cells produce interleukin-4 (IL-4), IL-5, and IL-13 from the type 2 gene cluster under regulation by transcription factors (TFs) including GATA3. To better understand transcriptional regulation of Th2 cell differentiation, we performed CRISPR-Cas9 screens targeting 1,131 TFs. We discovered that activity-dependent neuroprotector homeobox protein (ADNP) was indispensable for immune reactions to allergen. Mechanistically, ADNP performed a previously unappreciated role in gene activation, forming a critical bridge in the transition from pioneer TFs to chromatin remodeling by recruiting the helicase CHD4 and ATPase BRG1. Although GATA3 and AP-1 bound the type 2 cytokine locus in the absence of ADNP, they were unable to initiate histone acetylation or DNA accessibility, resulting in highly impaired type 2 cytokine expression. Our results demonstrate an important role for ADNP in promoting immune cell specialization.

Hematology, Biochemistry, and Protein Electrophoresis Reference Intervals of Western European Hedgehog (Erinaceus europaeus) from a Rehabilitation Center in Northern Portugal.

The Western European hedgehog (Erinaceus europaeus) can work as a bioindicator of environmental pollution and be a host for multiple zoonotic agents, making it relevant in terms of One Health studies. It is essential to deepen the knowledge on this species and calculate reference intervals (RIs) for the usual hematological and biochemical parameters. For this retrospective study (2017-2022), the archives of the Clinical Pathology Laboratory (LPC) of University of Trás-os-Montes and Alto Douro (UTAD) Veterinary Teaching Hospital were analyzed. Data of hematology, clinical biochemistry, and protein electrophoresis from 37 healthy hedgehogs of the Wild Animal Rehabilitation Center at UTAD, Northern Portugal, were included. It was possible to calculate RIs for almost all of the variables in the study, using Reference Value Advisor V2.1. Moreover, sex and age effects were investigated: alkaline phosphatase (p = 0.012, higher in males); total proteins (p = 0.034, higher in adults); mean cell volume (p = 0.007) and mean corpuscular hemoglobin (p = 0.010) (both higher in juveniles); and red blood cell distribution width (p = 0.021, higher in adults). Our study allowed for the first time to define RIs for a population of hedgehogs in Portugal, having a potentially relevant impact on species conservation and in the human-animal health interface.

An αvß3 integrin checkpoint is critical for efficient TH2 cell cytokine polarization and potentiation of antigen-specific immunity.

Naive CD4+ T lymphocytes initially undergo antigen-specific activation to promote a broad-spectrum response before adopting bespoke cytokine expression profiles shaped by intercellular microenvironmental cues, resulting in pathogen-focused modular cytokine responses. Interleukin (IL)-4-induced Gata3 upregulation is important for the helper type 2 T cell (TH2 cell) polarization associated with anti-helminth immunity and misdirected allergic inflammation. Whether additional microenvironmental factors participate is unclear. Using whole mouse-genome CRISPR-Cas9 screens, we discovered a previously unappreciated role for αvß3 integrin in TH2 cell differentiation. Low-level αvß3 expression by naive CD4+ T cells contributed to pan-T cell activation by promoting T-T cell clustering and IL-2/CD25/STAT5 signaling. Subsequently, IL-4/Gata3-induced selective upregulation of αvß3 licensed intercellular αvß3-Thy1 interactions among TH2 cells, enhanced mammalian target of rapamycin (mTOR) signaling, supported differentiation and promoted IL-5/IL-13 production. In mice, αvß3 was required for efficient, allergen-driven, antigen-specific lung TH2 cell responses. Thus, αvß3-expressing TH2 cells form multicellular factories to propagate and amplify TH2 cell responses.

Identification of aceNKPs, a committed common progenitor population of the ILC1 and NK cell continuum.

The development of innate lymphoid cell (ILC) transcription factor reporter mice has shown a previously unexpected complexity in ILC hematopoiesis. Using novel polychromic mice to achieve higher phenotypic resolution, we have characterized bone marrow progenitors that are committed to the group 1 ILC lineage. These common ILC1/NK cell progenitors (ILC1/NKP), which we call "aceNKPs", are defined as lineage-Id2+IL-7Rα+CD25-α4ß7-NKG2A/C/E+Bcl11b-. In vitro, aceNKPs differentiate into group 1 ILCs, including NK-like cells that express Eomes without the requirement for IL-15, and produce IFN-γ and perforin upon IL-15 stimulation. Following reconstitution of Rag2-/-Il2rg-/- hosts, aceNKPs give rise to a spectrum of mature ILC1/NK cells (regardless of their tissue location) that cannot be clearly segregated into the traditional ILC1 and NK subsets, suggesting that group 1 ILCs constitute a dynamic continuum of ILCs that can develop from a common progenitor. In addition, aceNKP-derived ILC1/NK cells effectively ameliorate tumor burden in a model of lung metastasis, where they acquired a cytotoxic NK cell phenotype. Our results identify the primary ILC1/NK progenitor that lacks ILC2 or ILC3 potential and is strictly committed to ILC1/NK cell production irrespective of tissue homing.

Correlation between changes in Timed Up and Go performance and cognition in older people with mild cognitive impairment: A longitudinal study.

BACKGROUND: Compare changes in performance on subtasks of the Timed Up and Go test over 32 months in older adults with and without mild cognitive impairment; analyze the correlation between frontal cognitive functions at baseline and changes in Timed Up and Go subtasks over time. METHODS: A longitudinal study was conducted involving 31 older adults (15 with and 16 without cognitive impairment). Functional mobility was assessed at both evaluations using an adapted version of the Timed Up and Go test and the Qualisys motion system. The test was divided into five subtasks: sit-to-stand, walking forward, turn-to-walk, walking back and turn-to-sit. Cognition was assessed at baseline using the Frontal Assessment Battery and the Clock Drawing Test. FINDINGS: Significant differences in changes in the sit-to-stand subtask over time were found between groups. The difference in the time required to conclude this subtask was greater in the older adults with cognitive impairment. However, the difference in kinematic variables (peak trunk speed and range of motion) was greater in the group without cognitive impairment. Strong and moderate correlations were found between frontal cognitive functions and changes in Timed Up and Go subtasks, especially those involving transitions (sit-to-stand, turn-to-walk and turn-to-sit) in both groups. INTERPRETATION: Motor intervention protocols should incorporate the sit-to-stand subtask in older adults with mild cognitive impairment. Moreover, changes in more complex subtasks seem to be related to frontal cognitive performance at baseline. Therefore, treatments that combine motor and cognitive functions should be administered to older adults regardless of cognitive impairment.

Prothrombin time and activated partial thromboplastin time reference intervals in dogs, by gender and age group using the Start®4 (Stago) / Intervalos de referência para o tempo de protrombina e tempo de tromboplastina parcial ativada no cão de acordo com a idade e o gênero utilizando o Start ® 4 (Stago)

Prothrombin time (PT) and the activated partial thromboplastin time (aPTT) are useful tools for the diagnosis and monitoring of coagulation disorders in Veterinary Medicine. Our objectives were: to establish reference intervals (RI) for PT and a PTT for the dog using the Start®4 (Stago), to compare the obtained RI with literature; to evaluate the effects of gender and age on the coagulation profile. Plasma samples of 122 healthy dogs (57 males; 65 females) aged between 4 months and 18 years, divided into three age groups (0-2 years old; 3-10 years old; > 10 years old) and grouped in to males and females were analysed. The RI were estimated following the ASVCP guidelines with the Reference Value Advisor software. The RI were: PT 6.7'' to 10.8''; aPTT 9.0'' to 14.8''. PT was significantly higher in females than in males. Dogs aged 10 years or older have significantly higher mean aPTT times than younger dogs. RI comparison showed a considerable percentage of cases outside the reference RI of the literature (PT - 79.3%; aPTT - 77.1%), demonstrating the need of each laboratory to calculate its own RI. The RI established in this study are applicable for the coagulation profile assessment in dogs.

O tempo de protrombina (TP) e o tempo de tromboplastina parcial ativada (TTPa) são ferramentas úteis para o diagnóstico e monitorização das alterações da coagulação em Medicina Veterinária. Os objetivos deste estudo foram: estabelecer intervalos de referência (IR) para TP e TTPa para o cão utilizando o Start®4 (Stago), de modo a comparar os IR obtidos com a literatura; avaliar os efeitos do sexo e da idade no perfil da coagulação. Foram usadas amostras de plasma de 122 cães saudáveis (57 machos; 65 fêmeas) com idades entre quatro meses e 18 anos, divididos em três grupos (0-2 anos; 3-10 anos; > 10 anos) e agrupados em machos e fêmeas. Os IR foram calculados seguindo as diretrizes da ASVCP com o software Reference Value Advisor. Os IR obtidos foram: PT 6,7 '' a 10,8 ''; TTPa 9,0 '' a 14,8 ''. O TP foi significativamente maior nas fêmeas do que nos machos. Os cães com 10 anos ou mais apresentaram tempos médios de TTPa significativamente maiores do que cães mais jovens. A comparação de IR mostrou uma percentagem considerável de casos fora do IR de referência da literatura (TP - 79,3%; TTPa - 77,1%), confirmando a necessidade de cada laboratório calcular seu próprio IR. Os IR estabelecidos neste estudo são aplicáveis na avaliação do perfil hemostático em cães.

Sono-Biosynthesis and Characterization of AuNPs from Danube Delta Nymphaea alba Root Extracts and Their Biological Properties.

Root extracts from Danube Delta Nymphaea alba were used to prepare gold nanoparticles (AuNPRn) by reducing HAuCl4 at different pHs (6.4-8.4) using ultrasonic irradiation: an easy, cheap, eco-friendly and green approach. Their antibacterial and anticancer activities were evaluated against Staphylococcus aureus and Escherichia coli, and A2780 ovarian cancer cells, respectively. The AuNPRn were characterized concerning their phytoconstituents (polyphenols, flavonoids and condensed tannins) and gold content. All of the nanoparticles were negatively charged. AuNPRn exhibited a hydrodynamic size distribution ranging from 32 nm to 280 nm, with the larger nanoparticles being obtained with an Au/root extract ratio of 0.56, pH 7 and 10 min of sonication (AuNPR1), whereas the smallest were obtained with an Au/root extract ratio of 0.24, pH 7.8 and 40 min of sonication (AuNPR4). The TEM/SEM images showed that the AuNPRn had different shapes. The ATR-FTIR indicated that AuNPRn interact mainly with hydroxyl groups present in the polyphenol compounds, which also confirm their high antioxidant capacity, except for AuNPR2 obtained at pH 6.4. Among the AuNPRn, the smallest ones exhibited enhanced antimicrobial and anticancer activities.

RORα is a critical checkpoint for T cell and ILC2 commitment in the embryonic thymus.

Type 2 innate lymphoid cells (ILC2) contribute to immune homeostasis, protective immunity and tissue repair. Here we demonstrate that functional ILC2 cells can arise in the embryonic thymus from shared T cell precursors, preceding the emergence of CD4+CD8+ (double-positive) T cells. Thymic ILC2 cells migrated to mucosal tissues, with colonization of the intestinal lamina propria. Expression of the transcription factor RORα repressed T cell development while promoting ILC2 development in the thymus. From RNA-seq, assay for transposase-accessible chromatin sequencing (ATAC-seq) and chromatin immunoprecipitation followed by sequencing (ChIP-seq) data, we propose a revised transcriptional circuit to explain the co-development of T cells and ILC2 cells from common progenitors in the thymus. When Notch signaling is present, BCL11B dampens Nfil3 and Id2 expression, permitting E protein-directed T cell commitment. However, concomitant expression of RORα overrides the repression of Nfil3 and Id2 repression, allowing ID2 to repress E proteins and promote ILC2 differentiation. Thus, we demonstrate that RORα expression represents a critical checkpoint at the bifurcation of the T cell and ILC2 lineages in the embryonic thymus.

THORACIC ORIGIN OF THE RIGHT RENAL ARTERY: AN INCIDENTAL FINDING.

Thoracic origin of a single renal artery with a normal position of the kidney is a very rare anatomical variant with just a dozen cases depicted in the medical literature. In this case report we describe an incidental finding of a main renal artery arising from the thoracic aorta at the 11th thoracic vertebral level in an asymptomatic 57 year-old man in a routine computed tomography (CT) on follow-up for chronic pancreatitis.

Influence of Ethanol on Emulsions Stabilized by Low Molecular Weight Surfactants.

The effect of ethanol on oil-in-water emulsions stabilized with low molecular weight surfactants was investigated. Oil-in-water emulsions were prepared containing varying percentages of ethanol and sunflower oil, and stabilized with different emulsifiers (Tween 20, Tween 80, and Lecithin). Droplet size, viscosity, density, and interfacial tension measurements were carried out. The droplet size of emulsions stabilized by each of the surfactants studied decreased with the addition of ethanol to the aqueous phase showing a minimum at a concentration of ethanol around 40%. The trend in droplet size is accompanied by a decrease in the interfacial tension between water and oil as the ethanol concentration increases. Viscosity measurements show that the change in viscosity of the final emulsion is the result of the change in viscosity of the continuous phase, as well as the change in solubility of the surfactants due to the addition of ethanol. The density of the continuous phase decreases with the addition of ethanol and it is possible to match the densities of the two phases in order to reduce the effect of creaming/sedimentation and improve stability. This study provides scientific evidence for the formulation of stable emulsions containing a range of ethanol form 0 to 40%. PRACTICAL APPLICATION: Formation and stability of food-grade emulsions in the presence of ethanol.

Polychromic Reporter Mice Reveal Unappreciated Innate Lymphoid Cell Progenitor Heterogeneity and Elusive ILC3 Progenitors in Bone Marrow.

Innate lymphoid cells (ILCs) play strategic roles in tissue homeostasis and immunity. ILCs arise from lymphoid progenitors undergoing lineage restriction and the development of specialized ILC subsets. We generated "5x polychromILC" transcription factor reporter mice to delineate ILC precursor states by revealing the multifaceted expression of key ILC-associated transcription factors (Id2, Bcl11b, Gata3, RORγt, and RORα) during ILC development in the bone marrow. This approach allowed previously unattained enrichment of rare progenitor subsets and revealed hitherto unappreciated ILC precursor heterogeneity. In vivo and in vitro assays identified precursors with potential to generate all ILC subsets and natural killer (NK) cells, and also permitted discrimination of elusive ILC3 bone marrow antecedents. Single-cell gene expression analysis identified a discrete ILC2-committed population and delineated transition states between early progenitors and a highly heterogeneous ILC1, ILC3, and NK precursor cell cluster. This diversity might facilitate greater lineage potential upon progenitor recruitment to peripheral tissues.