S100A9: The Unusual Suspect Connecting Viral Infection and Inflammation.

The study of S100A9 in viral infections has seen increased interest since the COVID-19 pandemic. S100A8/A9 levels were found to be correlated with the severity of COVID-19 disease, cytokine storm, and changes in myeloid cell subsets. These data led to the hypothesis that S100A8/A9 proteins might play an active role in COVID-19 pathogenesis. This review explores the structures and functions of S100A8/9 and the current knowledge on the involvement of S100A8/A9 and its constituents in viral infections. The potential roles of S100A9 in SARS-CoV-2 infections are also discussed.

From land to ocean: One month for southern elephant seal pups to acquire aquatic skills prior to their first departure to sea.

Weaned southern elephant seals (SES) quickly transition from terrestrial to aquatic life after a 5- to 6-week post-weaning period. At sea, juveniles and adult elephant seals present extreme, continuous diving behaviour. Previous studies have highlighted the importance of the post-weaning period for weanlings to prepare for the physiological challenges of their future sea life. However, very little is known about how their body condition during this period may influence the development of their behaviour and brain activities. To characterise changes in the behavioural and brain activity of weanlings prior to ocean departure, we implemented a multi-logger approach combining measurements of movements (related to behaviour), pressure (related to diving), and brain electrical activity. As pups age, the amount of time allocated to resting decreases in favour of physical activity. Most resting (9.6 ± 1.2 h/day) takes place during daytime, with periods of slow-wave sleep representing 4.9 ± 0.9 h/day during the first 2 weeks. Furthermore, an increasing proportion of physical activity transitions from land to shore. Additionally, pups in poorer condition (lean group) are more active earlier than those in better condition (corpulent group). Finally, at weaning, clear circadian activity with two peaks at dawn and dusk is observed, and this pattern remains unchanged during the 4 weeks on land. This circadian pattern matches the one observed in adults at sea, with more prey catches at dawn and dusk, raising the question of whether it is endogenous or triggered by the mother during lactation.

Breaking the fast: first report of dives and ingestion events in molting southern elephant seals.

Southern elephant seals (SES) experience a 'catastrophic molt', a costly event characterized by the renewal of both hair and epidermis that requires high peripheral vascular circulation. Molting animals are therefore constrained by high metabolic heat loss and are thought to fast and remain on land. To examine the ability of individuals to balance the energetic constraints of molting on land we investigate the stomach temperature and movement patterns of molting female SES. We find that 79% of females swam and 61% ingested water or prey items, despite the cost of cold-water exposure while molting. This behavior was related to periods of warm and low wind conditions, and females that dived and ingested more often, lost less body mass. We conclude that the paradigm of fasting during the molt in this species, and the fitness consequences of this behavior should be reconsidered, especially in the context of a changing climate.

An in vitro autologous, vascularized, and immunocompetent Tissue Engineered Skin model obtained by the self-assembled approach.

A complete in vitro skin model, containing resident cell types is needed to understand physiology and to consider the role of immune and endothelial cells in dermal drug testing. In this study, a cell extraction technique was developed to isolate resident skin cells from the same human donor while preserving the immune and endothelial cells. Then those cells were used to reconstruct an autologous, vascularized, and immunocompetent Tissue-Engineered Skin model, aviTES. Phenotypic characterization of the viable cells was performed on freshly isolated cells and after thawing through flow cytometry. Dermal cell extracts were characterized as fibroblasts, endothelial and immune cells, and the average amount of each cell type represents 4, 0.5, and 1 million viable cells per g of the dermis, respectively. The 3D models, TES and aviTES, were characterized by a fully differentiated epidermis that showed an increase in the presence of Ki67+ cells in the basolateral layer of the aviTES model. Capillary-like network formation, through the self-assembly of endothelial cells, and the presence of functional immune cells were identified through immunofluorescence staining in aviTES. In addition, the aviTES model was immunocompetent, as evidenced by its capacity to increase the production of pro-inflammatory cytokines TNF-α, MIP-1α, and GM-CSF following LPS stimulation. This study describes an autologous skin model containing a functional resident skin immune system and a capillary network. It provides a relevant tool to study the contribution of the immune system to skin diseases and inflammatory responses and to investigate resident skin cell interactions and drug development. STATEMENT OF SIGNIFICANCE: There is an urgent need for a complete in vitro skin model containing the resident cell types to better understand the role of immune and endothelial cells in skin and to be able to use it for drug testing. Actual 3D models of human skin most often contain only fibroblasts and keratinocytes with a limited number of models containing endothelial cells or a limited variety of immune cells. This study describes an autologous skin model containing a functional resident skin immune system and a capillary network. It provides a relevant tool to study the contribution of the immune system to skin diseases and inflammatory responses and to investigate interactions between resident skin cell, improving our capacity to develop new drugs.

Diving behaviour of southern elephant seals: new models of behavioural and ecophysiological adjustments of oxygen store management.

Among pinnipeds, southern elephant seals (SESs, Mirounga leonina) are extreme divers that dive deeply and continuously along foraging trips to restore their body stores after fasting on land during breeding or moulting. Their replenishment of body stores influences their energy expenditure during dives and their oxygen (O2) reserves (via muscular mass), yet how they manage their O2 stores during their dives is not fully understood. In this study, 63 female SESs from Kerguelen Island were equipped with accelerometers and time-depth recorders to investigate changes in diving parameters through their foraging trips. Two categories of dive behaviour were identified and related to the body size of individuals, with smaller SESs performing shallower and shorter dives requiring greater mean stroke amplitude compared with larger individuals. In relation to body size, the larger seals had lower estimated oxygen consumption levels for a given buoyancy (i.e. body density) compared with smaller individuals. However, both groups were estimated to have the same oxygen consumption of 0.079±0.001 ml O2 stroke-1 kg-1 for a given dive duration and at neutral buoyancy when the cost of transport was minimal. Based on these relationships, we built two models that estimate changes in oxygen consumption according to dive duration and body density. The study highlights that replenishing body stores improves SES foraging efficiency, as indicated by increased time spent at the bottom of the ocean. Thus, prey-capture attempts increase as SES buoyancy approaches the neutral buoyancy point.

Immune Cells Release MicroRNA-155 Enriched Extracellular Vesicles That Promote HIV-1 Infection.

The hallmark of HIV-1 infection is the rapid dysregulation of immune functions. Recent investigations for biomarkers of such dysregulation in people living with HIV (PLWH) reveal a strong correlation between viral rebound and immune activation with an increased abundance of extracellular vesicles (EVs) enriched with microRNA-155. We propose that the activation of peripheral blood mononuclear cells (PBMCs) leads to an increased miR-155 expression and production of miR-155-rich extracellular vesicles (miR-155-rich EVs), which can exacerbate HIV-1 infection by promoting viral replication. PBMCs were incubated with either HIV-1 (NL4.3Balenv), a TLR-7/8 agonist, or TNF. EVs were harvested from the cell culture supernatant by differential centrifugation, and RT-qPCR quantified miR-155 in cells and derived EVs. The effect of miR-155-rich EVs on replication of HIV-1 in incubated PBMCs was then measured by viral RNA and DNA quantification. HIV-1, TLR7/8 agonist, and TNF each induced the release of miR-155-rich EVs by PBMCs. These miR-155-rich EVs increased viral replication in PBMCs infected in vitro. Infection with HIV-1 and inflammation promote the production of miR-155-rich EVs, enhancing viral replication. Such autocrine loops, therefore, could influence the course of HIV-1 infection by promoting viral replication.

HIV Replication Increases the Mitochondrial DNA Content of Plasma Extracellular Vesicles.

Extracellular vesicles (EVs) and their cargo have been studied intensively as potential sources of biomarkers in HIV infection; however, their DNA content, particularly the mitochondrial portion (mtDNA), remains largely unexplored. It is well known that human immunodeficiency virus (HIV) infection and prolonged antiretroviral therapy (ART) lead to mitochondrial dysfunction and reduced mtDNA copy in cells and tissues. Moreover, mtDNA is a well-known damage-associated molecular pattern molecule that could potentially contribute to increased immune activation, oxidative stress, and inflammatory response. We investigated the mtDNA content of large and small plasma EVs in persons living with HIV (PLWH) and its implications for viral replication, ART use, and immune status. Venous blood was collected from 196 PLWH, ART-treated or ART-naïve (66 with ongoing viral replication, ≥20 copies/mL), and from 53 HIV-negative persons, all recruited at five HIV testing or treatment centers in Burkina Faso. Large and small plasma EVs were purified and counted, and mtDNA level was measured by RT-qPCR. Regardless of HIV status, mtDNA was more abundant in large than small EVs. It was more abundant in EVs of viremic than aviremic and control participants and tended to be more abundant in participants treated with Tenofovir compared with Zidovudine. When ART treatment was longer than six months and viremia was undetectable, no variation in EV mtDNA content versus CD4 and CD8 count or CD4/CD8 ratio was observed. However, mtDNA in large and small EVs decreased with years of HIV infection and ART. Our results highlight the impact of viral replication and ART on large and small EVs' mtDNA content. The mechanisms underlying the differential incorporation of mtDNA into EVs and their effects on the surrounding cells warrant further investigation.

Canine Model of Human Frailty: Adaptation of a Frailty Phenotype in Older Dogs.

Frailty is a clinical presentation resulting from age-related cumulative declines in several physiological systems. The aim of this study was to adapt the concept of frailty to the domestic dog, as a model for frailty research, by characterizing a 5-criterion frailty phenotype using objective measurement, and to investigate its independent association with death. A prospective cohort including 80 Labrador and Golden Retriever dogs aged 9 years or older was conducted between March 2015 and July 2020. An adapted frailty phenotype was defined according to the presence of 5 criteria (weakness, slowness, poor endurance, low physical activity, and shrinking) evaluated at baseline from physical performance tests and items from questionnaire and physical examination. Survival analysis was used to investigate the association between frailty status and time to all-cause death over 5 years of follow-up. Frailty status was significantly associated with all-cause death, with median survival times of 10.5 months, 35.4 months, and 42.5 months, respectively for dogs with 3 or more criteria (frail dogs), dogs with 1 or 2 criteria (prefrail dogs), and nonfrail dogs. Independently of age, sex, breed, sterilization, and sex-sterilization interaction, frail dogs died significantly faster than nonfrail dogs at baseline (adjusted hazard ratio = 5.86; 95% confidence interval = 2.45-14.0; p < .01). This significant association persisted after controlling for other potential confounders. Frailty, assessed by a 5-criterion phenotype, was predictive of all-cause death, in geriatric Labrador and Golden Retriever dogs. The concept of frailty seems adaptable to the dog.

Remote Medical Scent Detection of Cancer and Infectious Diseases With Dogs and Rats: A Systematic Review.

BACKGROUND: Remote medical scent detection of cancer and infectious diseases with dogs and rats has been an increasing field of research these last 20 years. If validated, the possibility of implementing such a technique in the clinic raises many hopes. This systematic review was performed to determine the evidence and performance of such methods and assess their potential relevance in the clinic. METHODS: Pubmed and Web of Science databases were independently searched based on PRISMA standards between 01/01/2000 and 01/05/2021. We included studies aiming at detecting cancers and infectious diseases affecting humans with dogs or rats. We excluded studies using other animals, studies aiming to detect agricultural diseases, diseases affecting animals, and others such as diabetes and neurodegenerative diseases. Only original articles were included. Data about patients' selection, samples, animal characteristics, animal training, testing configurations, and performances were recorded. RESULTS: A total of 62 studies were included. Sensitivity and specificity varied a lot among studies: While some publications report low sensitivities of 0.17 and specificities around 0.29, others achieve rates of 1 sensitivity and specificity. Only 6 studies were evaluated in a double-blind screening-like situation. In general, the risk of performance bias was high in most evaluated studies, and the quality of the evidence found was low. CONCLUSIONS: Medical detection using animals' sense of smell lacks evidence and performances so far to be applied in the clinic. What odors the animals detect is not well understood. Further research should be conducted, focusing on patient selection, samples (choice of materials, standardization), and testing conditions. Interpolations of such results to free running detection (direct contact with humans) should be taken with extreme caution. Considering this synthesis, we discuss the challenges and highlight the excellent odor detection threshold exhibited by animals which represents a potential opportunity to develop an accessible and non-invasive method for disease detection.

How's my kitty? Acoustic parameters of cat-directed speech in human-cat interactions.

In Western cultures, humans tend to use a specific kind of speech when talking to their pets, characterised, from an acoustical point of view, by elevated pitch and greater pitch modulation. Pet-directed speech (PDS), which has been mainly studied in dogs, shares some acoustic features with infant-directed speech (IDS), used when talking to young children. The purpose of this study was to test the hypothesis that adult humans also modify characteristics of their voice when talking to a cat. We compared acoustic parameters of speech directed to cats (CDS) and speech directed to adult humans (ADS). In a first experiment, we compared ADS and CDS utterances of male and female participants, addressing cats through video recordings, under controlled laboratory conditions. Both men and women used a higher pitch (mean fundamental frequency, or mean F0) in CDS vs. ADS. The second experiment was conducted under conditions allowing direct cat-human interactions, in a cohort of women. Once again, mean F0 was significantly higher in CDS vs. ADS. Overall, these data confirm our hypothesis that humans change the way they speak when addressing a cat, mainly by increasing the pitch of their voice. Further research is needed to fully investigate specificities of this speech.

Plasma vesicular miR-155 as a biomarker of immune activation in antiretroviral treated people living with HIV.

People living with HIV (PLWH), despite suppression of viral replication with antiretroviral therapy (ART), have high morbidity and mortality due to immune activation and chronic inflammation. Discovering new biomarkers of immune activation status under ART will be pertinent to improve PLWH quality of life when the majority will be treated. We stipulate that plasma large and small extracellular vesicle (EVs) and their microRNA content could be easily measured biomarkers to monitor immune activation in PLWH. Venous blood samples from n = 128 ART-treated PLWH with suppressed viral load (≤ 20 copies/mL) and n = 60 HIV-uninfected participants were collected at five testing or treatment centers of PLWH in Burkina Faso. Large and small plasma EVs were purified, counted, and the mature miRNAs miR-29a, miR-146a, and miR-155 were quantified by RT-qPCR. Diagnostic performances of large and small EVs miRNAs level were evaluated by receiver operating characteristic (ROC) curve analysis and principal component analysis (PCA). Among the EVs microRNA measured, only large EVs miR-155 copies distinguished PLWH with immune activation, with AUC of 0.75 for CD4/CD8 < 1 (95% CI: 0.58-0.91, P = 0.0212), and 0.77 for CD8 T cells ≥ 500/µL (95% CI: 0.63-0.92, P = 0.0096). In addition, PCA results suggest that large EVs miR-155 copies may be a biomarker of immune activation. Since miR-155 may influence immune cell function, its enrichment in large EV subpopulations could be a functional biomarker of immune activation in PLWH on ART. This measure could help to monitor and diagnose the immune activation with more accuracy.

Sticking Together: Energetic Consequences of Huddling Behavior in Hibernating Juvenile Garden Dormice.

AbstractHibernation, or multiday torpor, allows individuals to save energy via substantial reductions of metabolism and body temperature but is regularly interrupted by euthermic phases called arousals. Social thermoregulation, or "huddling," can act in synergy with torpor in reducing individuals' energy and heat losses. In the wild, the garden dormouse (Eliomys quercinus) combines both strategies, which are crucial for winter survival of juveniles with limited prehibernation body fat reserves. We investigated via thermographic and temperature measurements (i) the energetic impact of huddling during an arousal from deep torpor, (ii) the dynamics of huddling behavior during hibernation, and (iii) its consequences during the entire winter in juvenile garden dormice. Thermographic images revealed a significant effect of huddling on torpor energetics, as it reduced heat exchange and mass loss by two-thirds in huddling versus single individuals during arousal. Our investigation of the dynamics of huddling further revealed a "random-like mechanistic" behavior during winter hibernation, as arousals from torpor were not always initiated by the same individuals. Animals took turns in initiating rewarming within a group, and the individual with highest body temperature during arousal entered into torpor later than the others within the huddle. The animals share both costs and benefits of huddling during arousals, without any energetic benefit of huddling over the entire winter on an individual level. We conclude that the dynamics of social thermoregulation during hibernation seems to counterbalance its benefit of reducing energetic costs associated against the energy-demanding process of rewarming from torpor.

Vesicular MicroRNA as Potential Biomarkers of Viral Rebound.

Changes in the cellular microRNA (miRNA) expression profile in response to HIV infection, replication or latency have been reported. Nevertheless, little is known concerning the abundance of miRNA in extracellular vesicles (EVs). In the search for a reliable predictor of viral rebound, we quantified the amount of miR-29a, miR-146a, and miR-155 in two types of plasma extracellular vesicles. Venous blood was collected from 235 ART-treated and ART-naive persons living with HIV (85 with ongoing viral replication, ≥20 copies/mL) and 60 HIV-negative participants at five HIV testing or treatment centers in Burkina Faso. Large and small plasma EVs were purified and counted, and mature miRNA miR-29a, miR-146a, and miR-155 were measured by RT-qPCR. Diagnostic performance of miRNA levels in large and small EVs was evaluated by a receiver operating characteristic curve analysis. The median duration of HIV infection was 36 months (IQR 14-117). The median duration of ART was 34 months (IQR 13-85). The virus was undetectable in 63.8% of these persons. In the others, viral load ranged from 108 to 33,978 copies/mL (median = 30,032). Large EVs were more abundant in viremic participants than aviremic. All three miRNAs were significantly more abundant in small EVs in persons with detectable HIV RNA, and their expression levels in copies per vesicle were a more reliable indicator of viral replication in ART-treated patients with low viremia (20-1000 copies/mL). HIV replication increased the production of large EVs more than small EVs. Combined with viral load measurement, quantifying EV-associated miRNA abundance relative to the number of vesicles provides a more reliable marker of the viral status. The expression level as copies per small vesicle could predict the viral rebound in ART-treated patients with undetectable viral loads.

Metabolic heat loss in southern elephant seals (Mirounga leonina) differs with stage of moult and between habitats.

The moult in southern elephant seals (Mirounga leonina) represents an especially energetically demanding period during which seals must maintain high skin temperature to facilitate complete replacement of body fur and upper dermis. In this study, heat flux from the body surface was measured on 18 moulting southern elephant seals to estimate metabolic heat loss in three different habitats (beach, wallow and vegetation). Temperature data loggers were also deployed on 10 southern elephant seals to monitor skin surface temperature. On average, heat loss of animals on the beach was greater than in wallows or vegetation, and greater in wallows than in vegetation. Heat loss across all habitats during the moult equated to 1.8 x resting metabolic rate (RMR). The greatest heat loss of animals was recorded in the beach habitat during the late moult, that represented 2.3 x RMR. Mass loss was 3.6 ± 0.3 kg day-1, resulting in changes in body condition as the moult progressed. As body condition declined, skin surface temperature also decreased, suggesting that as animals approached the end of the moult blood flow to the skin surface was no longer required for hair growth.

Immunovirological discordance among female sex workers who start antiretroviral therapy in Burkina Faso.

BACKGROUND: In people living with HIV/AIDS (PLWHA), initiation of antiretroviral therapy (ART) leads to sustained effective suppression of viral replication and increasing CD4 + T cell count. However, a fraction of ART-treated patients still fail to reach adequate CD4 + T cell number despite a suppressed viral load (VL), and this phenomenon is defined as immunovirological discordance (IVD). In Africa, several studies have reported immunovirological outcomes of antiretroviral therapy, but little is known about IVD occurrence in Female sex workers (FSW). This study aimed to assess the prevalence of IVD and associated factors among a cohort of HIV infected FSW in Burkina Faso. METHODS: We conducted a cohort study from December 2003 to October 2016. Immunovirological discordance was defined as CD4 + T cell gain < 100 cells/µL despite a suppressed VL (VL < 1000 copies/mL) 12 months after ART initiation. The CD4 + T cells were counted using BD FACSCount™ System and point of care Pima™ CD4 + Analyzer. HIV-1 RNA was quantified by real-time polymerase-chain-reaction assay with the use of the ABI 7000 system. We conducted a logistic regression to identify factors associated with discordant responses. RESULTS: Among the 123 HIV-1 infected FSW having at least 12 months follow-up on ART, 105 (85.4%) achieved HIV-1 RNA suppression. Among the latter 25 gained less than 100 CD4 + T cells within 12 months follow-up. The IVD rate was 23.8% (95%CI 16.04%-33.11%). After adjustment for age, WHO clinical stage and ART regimen including nucleoside/nucleotide reverse transcriptase inhibitors, only baseline CD4 + T cell count between 200 to 350 cells/µL (adjusted OR: 4.15; 95%CI 1.13-15.22) and 350 to 500 cells/µL (adjusted OR: 17.50; 95%CI 2.68-114.31) remain significantly associated with IVD occurrence. CONCLUSIONS: Immunovirological discordance response was common in FSW with proportions close to those observed in the general population. A diagnosis and personalized follow-up of patients who do not achieve full immune reconstitution would make it possible to avoid complications in terms of morbidity and mortality.

Functional Role of AGAP2/PIKE-A in Fcγ Receptor-Mediated Phagocytosis.

In phagocytes, cytoskeletal and membrane remodeling is finely regulated at the phagocytic cup. Various smaFll G proteins, including those of the Arf family, control these dynamic processes. Human neutrophils express AGAP2, an Arf GTPase activating protein (ArfGAP) that regulates endosomal trafficking and focal adhesion remodeling. We first examined the impact of AGAP2 on phagocytosis in CHO cells stably expressing the FcγRIIA receptor (CHO-IIA). In unstimulated CHO-IIA cells, AGAP2 only partially co-localized with cytoskeletal elements and intracellular compartments. In CHO-IIA cells, AGAP2 transiently accumulated at actin-rich phagocytic cups and increased Fcγ receptor-mediated phagocytosis. Enhanced phagocytosis was not dependent on the N-terminal GTP-binding protein-like (GLD) domain of AGAP2. AGAP2 deleted of its GTPase-activating protein (GAP) domain was not recruited to phagocytic cups and did not enhance the engulfment of IgG-opsonized beads. However, the GAP-deficient [R618K]AGAP2 transiently localized at the phagocytic cups and enhanced phagocytosis. In PLB-985 cells differentiated towards a neutrophil-like phenotype, silencing of AGAP2 reduced phagocytosis of opsonized zymosan. In human neutrophils, opsonized zymosan or monosodium urate crystals induced AGAP2 phosphorylation. The data indicate that particulate agonists induce AGAP2 phosphorylation in neutrophils. This study highlights the role of AGAP2 and its GAP domain but not GAP activity in FcγR-dependent uptake of opsonized particles.

Heads and Tails: An Analysis of Visual Signals in Cats, Felis catus.

Visual communication involves specific signals. These include the different positions of mobile body elements. We analyzed visual configurations in cats that involve ears and the tail. We aimed at deciphering which features of these configurations were the most important in cats' interactions with other cats and with humans. We observed a total of 254 cat-cat interactions within a sample of 29 cats, during a total of 100 h of observation scheduled with the "Behavioral dependent onset of sampling" method and using the "All occurences" sampling method. In addition, we sampled 10 interactions between cats and humans. In cat-cat interactions, we noted the positions of ears and tail of both protagonists, as well as the outcome of the interaction, which was either positive/neutral or negative. In a great majority of the 254 interactions sampled, both cats held their tail down. On the contrary, ear position was a critical element in predicting the outcome. When both partners held their ears erect, the outcome was significantly positive, such as rubbing or close proximity. In all other cases of the position of ears in both cats, the outcome was negative, with increased distance of the partners. Although the tail did not seem to play a significant role in visual configurations in cat interactions, the "tail-up" display was important when a cat approached a human being. In the vast majority of cases the cat rubbed itself on a human's leg(s). Thus, we may conclude that the presence of a human has a specific meaning in the cat's world, probably as the result of a long period of commensalism. It is important for pet owners to understand the signals that cats use with other cats and with humans in order to promote the welfare of cats.

Effects of a new dietary supplement on behavioural responses of dogs exposed to mild stressors.

BACKGROUND & OBJECTIVES: The effectiveness of a new dietary supplement (derived from fish hydrolysate and melon juice concentrate rich in superoxide dismutase) in reducing fear and stress-related behaviours in pet dogs was examined in a double-blind, placebo-controlled, randomized study. METHODS: 39 dogs were recruited after the owners had filled out a fear susceptibility index questionnaire. Over a 30-day period, one group of dogs received the supplement, and another group a placebo. Twelve behavioural variables were recorded in a series of four subtests (ST1-ST4) on days 0, 15 and 30. Saliva cortisol levels were measured before and after each set of STs. RESULTS: The dogs rated as more fearful displayed significantly higher cortisol values before the day 0 test session, were less active, spent less time playing with the experimenter, and approached the unfamiliar object less frequently. The owners did not correctly guess whether their dog had received the supplement or not. Behaviours of dogs were significantly different across the three sessions, with significant increases of stress-related behaviours (time spent in the door zone, number of interactions with the door, of whining, and of lip-licking). Conversely time spent with the experimenter increased, interactions and curiosity for the novel object and play with the experimenter decreased, presumably due to a habituation process. This suggests that the design of the four subtests session was relevant to test for mild stressors situations. Moreover, supplemented and placebo dogs responded differently to the three test sessions, indicating a supplement effect on dogs' behaviours and their adaptation to mild stressors situations. CONCLUSION: The trial results suggested that the supplement facilitates activity and curiosity in a familiar environment, promotes dog-human interactions with an increased human familiarity, and tends to reduce subtle stress behaviours. Our results suggest that the supplement was effective in the context of mild stressors and habituation.

Proliferation of peripheral blood mononuclear cells from healthy piglets after mitogen stimulation as indicators of disease resilience.

Disease resilience refers to the productivity of an animal under disease. Given the high biosecurity of pig nucleus herds, traits that can be measured on healthy pigs and that are genetically correlated with disease resilience, that is, genetic indicator traits, offer a strategy to select for disease resilience. Our objective was to evaluate mitogen stimulation assays (MSAs) on peripheral blood mononuclear cells (PBMCs) from young healthy pigs as genetic indicators for disease resilience. Data were from a natural disease challenge in which batches of 60 or 75 naïve Yorkshire × Landrace piglets were introduced every 3 wk into a continuous flow barn that was seeded with multiple diseases. In this environment, disease resilience traits, including growth, treatment, and mortality rates, were recorded on 3,136 pigs that were genotyped with a high-density marker panel. PBMCs from 882 of these pigs from 19 batches were isolated from whole blood collected prior to the disease challenge and stimulated with five mitogens: concanavalin A (ConA), phytohemagglutinin (PHA), pokeweed mitogen (PWM), lipopolysaccharide (LPS), and phorbol myristate acetate (PMA). The proliferation of cells was evaluated at 48, 72, and 96 h and compared with unstimulated samples (rest count). Heritabilities of cell proliferation were estimated using a model with batch as a fixed effect and covariates of entry age; rest count; complete blood count proportions of lymphocytes, monocytes, eosinophils, and basophils; and pen, litter, and animal genetics as random effects. Heritability estimates were highest for response to ConA (0.30 ± 0.09, 0.28 ± 0.10, 0.17 ± 0.10, and 0.25 ±0.10 at 48, 72, and 96 h after stimulation and for area under the curve across the three time points, respectively). Estimates were in a similar range for response to PHA and PMA but low for PWM and LPS. Responses to ConA, PHA, and PMA were moderately genetically correlated with several disease resilience traits and in the expected direction, but individual estimates were not significantly different from zero due to large SEs. In conclusion, although validation is needed, MSAss, in particular based on ConA, show promise as genetic indicator traits for disease resilience.

Plasma Extracellular Vesicle Subtypes May be Useful as Potential Biomarkers of Immune Activation in People With HIV.

BACKGROUND: Extracellular vesicles (EVs) are intercellular messengers with epigenetic potential since they can shuttle microRNA (miRNA). EVs and miRNA play a role in human immunodeficiency virus (HIV) infection immunopathogenesis. Chronic immune activation and systemic inflammation during HIV infection despite effective antiretroviral therapy (ART) are associated with non-acquired immunodeficiency syndrome (AIDS) comorbidities in people living with HIV (PLWH). Analysis of plasma EVs and their miRNA content may be useful as immune activation or inflammatory biomarkers in PLWH receiving ART. In this study, we hypothesized that the number, size, and miRNA of large and small EVs could reflect immune activation associated with an elevated CD8 T-cell count or a low CD4/CD8 ratio in PLWH. METHODS: Plasma EVs subtype purified from PLWH and uninfected controls were sized using dynamic light scattering and quantified using flow cytometry and acetylcholine esterase (AChE) activity. Expression of mature miRNAs miR-92, miR-155, miR-223 was measured by quantitative reverse-transcriptase polymerase chain reaction in EVs and leucocytes. RESULTS: HIV infection induces increased production of small EVs in plasma. EV subtypes were differentially enriched in miR-92, miR-155, and miR-223. Positive correlations between CD8 T-cell count and large EVs abundance and small EVs AChE activity were observed. CD4/CD8 ratio was negatively correlated with small EV AChE activity, and miRNA-155 level per small EV was negatively correlated with CD8 T-cell count. CONCLUSIONS: These findings suggest that quantifying large or small EVs and profiling miRNA content per EV might provide new functional biomarkers of immune activation and inflammation.