ABSTRACT
Metformin, the world's most prescribed anti-diabetic drug, is also effective in preventing type 2 diabetes in people at high risk1,2. More than 60% of this effect is attributable to the ability of metformin to lower body weight in a sustained manner3. The molecular mechanisms by which metformin lowers body weight are unknown. Here we show-in two independent randomized controlled clinical trials-that metformin increases circulating levels of the peptide hormone growth/differentiation factor 15 (GDF15), which has been shown to reduce food intake and lower body weight through a brain-stem-restricted receptor. In wild-type mice, oral metformin increased circulating GDF15, with GDF15 expression increasing predominantly in the distal intestine and the kidney. Metformin prevented weight gain in response to a high-fat diet in wild-type mice but not in mice lacking GDF15 or its receptor GDNF family receptor α-like (GFRAL). In obese mice on a high-fat diet, the effects of metformin to reduce body weight were reversed by a GFRAL-antagonist antibody. Metformin had effects on both energy intake and energy expenditure that were dependent on GDF15, but retained its ability to lower circulating glucose levels in the absence of GDF15 activity. In summary, metformin elevates circulating levels of GDF15, which is necessary to obtain its beneficial effects on energy balance and body weight, major contributors to its action as a chemopreventive agent.
Subject(s)
Body Weight/drug effects , Energy Metabolism/drug effects , Growth Differentiation Factor 15/metabolism , Metformin/pharmacology , Administration, Oral , Adult , Aged , Animals , Blood Glucose/analysis , Blood Glucose/metabolism , Diet, High-Fat , Double-Blind Method , Energy Intake/drug effects , Enterocytes/cytology , Enterocytes/drug effects , Female , Glial Cell Line-Derived Neurotrophic Factor Receptors/antagonists & inhibitors , Glial Cell Line-Derived Neurotrophic Factor Receptors/deficiency , Glial Cell Line-Derived Neurotrophic Factor Receptors/genetics , Growth Differentiation Factor 15/blood , Growth Differentiation Factor 15/deficiency , Growth Differentiation Factor 15/genetics , Homeostasis/drug effects , Humans , Intestines/cytology , Intestines/drug effects , Male , Metformin/administration & dosage , Mice , Mice, Obese , Middle Aged , Weight Loss/drug effectsABSTRACT
An Amendment to this paper has been published and can be accessed via a link at the top of the paper.
ABSTRACT
The maximum rate at which animals take up oxygen from their environment (MO2,max) is a crucial aspect of their physiology and ecology. In fishes, MO2,max is commonly quantified by measuring oxygen uptake either during incremental swimming tests or during recovery from an exhaustive chase. In this Commentary, we compile recent studies that apply both techniques to the same fish and show that the two methods typically yield different mean estimates of MO2,max for a group of individuals. Furthermore, within a group of fish, estimates of MO2,max determined during swimming are poorly correlated with estimates determined during recovery from chasing (i.e. an individual's MO2,max is not repeatable across methods). One explanation for the lack of agreement is that these methods measure different physiological states, each with their own behavioural, anatomical and biochemical determinants. We propose that these methods are not directly interchangeable but, rather, each is suited to address different questions in fish biology. We suggest that researchers select the method that reflects the biological contexts of their study, and we advocate for the use of accurate terminology that acknowledges the technique used to elevate MO2 (e.g. peak MO2,swim or peak MO2,recovery). If the study's objective is to estimate the 'true' MO2,max of an individual or species, we recommend that pilot studies compare methods, preferably using repeated-measures designs. We hope that these recommendations contribute new insights into the causes and consequences of variation in MO2,max within and among fish species.
Subject(s)
Fishes , Oxygen Consumption , Swimming , Animals , Swimming/physiology , Fishes/physiology , Fishes/metabolism , Oxygen Consumption/physiology , Oxygen/metabolismABSTRACT
Rift Valley fever virus (RVFV) is a mosquito-borne RNA virus of the Phlebovirus genus in the phenuviridae family. Its genome is trisegmented with small (S), medium (M) and large (L) fragments. In nature, the virus exists as a single serotype that is responsible for outbreaks of Rift Valley fever (RVF), a zoonotic disease that often occurs in Africa and the Middle East. RVFV genomes are thought to undergo both recombination and reassortment and investigations of these events is important for monitoring the emergence of virulent strains and understanding the evolutionary characteristics of this virus. The aim of this study was to characterize the genomes of RVFV isolates from cattle, sheep, and goats collected during an interepidemic period in Kenya between June 2016 and November 2021. A total of 691 serum samples from cattle (n = 144), goats (n = 185) and sheep (n = 362) were analysed at the Central Veterinary Laboratories. The competitive IgM-capture ELISA, was used to screen the samples; 205 samples (29.67%) tested positive for RVFV. Of the 205 positive samples, 42 (20.5%) were from cattle, 57 (27.8%) from goats, and 106 (51.7%) from sheep. All the IgM-positive samples were further analyzed by qPCR, and 24 (11.71%) tested positive with Ct values ranging from 14.788 to 38.286. Two samples, 201808HABDVS from sheep and 201810CML3DVS from cattle, had Ct values of less than 20.0 and yielded whole genome sequences with 96.8 and 96.4 coverage, respectively. There was no statistically significant evidence of recombination in any of the three segments and also phylogenetic analysis showed no evidence of reassortment in the two isolated RVFV segments when compared with other isolates of different lineages from previous outbreaks whose genomes are deposited in the GenBank. No evidence of reassortment leaves room for other factors to be the most probable contributors of change in virulence, pathogenicity and emergence of highly virulent strains of the RVFV.
Subject(s)
Cattle Diseases , Genome, Viral , Goat Diseases , Goats , Phylogeny , Rift Valley Fever , Rift Valley fever virus , Sheep Diseases , Animals , Goats/virology , Rift Valley fever virus/genetics , Rift Valley fever virus/isolation & purification , Sheep , Rift Valley Fever/virology , Rift Valley Fever/epidemiology , Cattle , Kenya/epidemiology , Goat Diseases/virology , Goat Diseases/epidemiology , Sheep Diseases/virology , Sheep Diseases/epidemiology , Cattle Diseases/virology , Cattle Diseases/epidemiology , Disease Outbreaks/veterinaryABSTRACT
The maximum rate at which fish can take up oxygen from their environment to fuel aerobic metabolism is an important feature of their physiology and ecology. Methods to quantify maximum oxygen uptake rate (MO2), therefore, should reliably and reproducibly estimate the highest possible MO2 by an individual or species under a given set of conditions (peak MO2). This study determined peak MO2 and its repeatability in Gulf killifish, Fundulus grandis, subjected to three methods to elevate metabolism: swimming at increasing water speeds, during recovery after an exhaustive chase, and after ingestion of a large meal. Estimates of peak MO2 during swimming and after an exhaustive chase were repeatable across two trials, whereas peak MO2 after feeding was not. Peak MO2 determined by the three methods was significantly different from one another, being highest during swimming, lowest after an exhaustive chase, and intermediate after feeding. In addition, peak MO2 during recovery from an exhaustive chase depended on the length of time of recovery: in nearly 60% of the trials, values within the first hour of the chase were lower than those measured later. A novel and important finding was that an individual's peak MO2 was not repeatable when compared across methods. Therefore, the peak MO2 estimated for a group of fish, as well as the ranking of individual MO2 within that group, depends on the method used to elevate aerobic metabolism.
Subject(s)
Fundulidae , Oxygen Consumption , Swimming , Animals , Fundulidae/physiology , Fundulidae/metabolism , Reproducibility of Results , Oxygen/metabolismABSTRACT
Under homeostatic conditions, animals use well-defined hypothalamic neural circuits to help maintain stable body weight, by integrating metabolic and hormonal signals from the periphery to balance food consumption and energy expenditure. In stressed or disease conditions, however, animals use alternative neuronal pathways to adapt to the metabolic challenges of altered energy demand. Recent studies have identified brain areas outside the hypothalamus that are activated under these 'non-homeostatic' conditions, but the molecular nature of the peripheral signals and brain-localized receptors that activate these circuits remains elusive. Here we identify glial cell-derived neurotrophic factor (GDNF) receptor alpha-like (GFRAL) as a brainstem-restricted receptor for growth and differentiation factor 15 (GDF15). GDF15 regulates food intake, energy expenditure and body weight in response to metabolic and toxin-induced stresses; we show that Gfral knockout mice are hyperphagic under stressed conditions and are resistant to chemotherapy-induced anorexia and body weight loss. GDF15 activates GFRAL-expressing neurons localized exclusively in the area postrema and nucleus tractus solitarius of the mouse brainstem. It then triggers the activation of neurons localized within the parabrachial nucleus and central amygdala, which constitute part of the 'emergency circuit' that shapes feeding responses to stressful conditions. GDF15 levels increase in response to tissue stress and injury, and elevated levels are associated with body weight loss in numerous chronic human diseases. By isolating GFRAL as the receptor for GDF15-induced anorexia and weight loss, we identify a mechanistic basis for the non-homeostatic regulation of neural circuitry by a peripheral signal associated with tissue damage and stress. These findings provide opportunities to develop therapeutic agents for the treatment of disorders with altered energy demand.
Subject(s)
Body Weight/physiology , Brain Stem/metabolism , Glial Cell Line-Derived Neurotrophic Factor Receptors/metabolism , Growth Differentiation Factor 15/metabolism , Animals , Brain Stem/cytology , Brain Stem/drug effects , Central Amygdaloid Nucleus/cytology , Central Amygdaloid Nucleus/physiology , Eating/physiology , Energy Metabolism/physiology , Feeding Behavior , Female , Glial Cell Line-Derived Neurotrophic Factor Receptors/deficiency , Glial Cell Line-Derived Neurotrophic Factor Receptors/genetics , Growth Differentiation Factor 15/genetics , Growth Differentiation Factor 15/pharmacology , Homeostasis , Male , Mice , Mice, Knockout , Neurons/drug effects , Neurons/metabolism , Parabrachial Nucleus/cytology , Parabrachial Nucleus/physiology , Stress, PsychologicalABSTRACT
This corrects the article DOI: 10.1038/nature24042.
ABSTRACT
This study asked whether interindividual variation in maximum and standard aerobic metabolic rates of the Gulf killifish, Fundulus grandis, correlates with gill morphology and cardiac mitochondrial bioenergetics, traits reflecting critical steps in the O2 transport cascade from the environment to the tissues. Maximum metabolic rate (MMR) was positively related to body mass, total gill filament length and myocardial oxygen consumption during maximum oxidative phosphorylation (multiple R2=0.836). Standard metabolic rate (SMR) was positively related to body mass, total gill filament length and myocardial oxygen consumption during maximum electron transport system activity (multiple R2=0.717). After controlling for body mass, individuals with longer gill filaments, summed over all gill arches, or greater cardiac respiratory capacity had higher whole-animal metabolic rates. The overall model fit and the explanatory power of individual predictor variables were better for MMR than for SMR, suggesting that gill morphology and myocardial bioenergetics are more important in determining active rather than resting metabolism. After accounting for body mass, heart ventricle mass was not related to variation in MMR or SMR, indicating that the quality of the heart (i.e. the capacity for mitochondrial metabolism) was more influential than heart size. Finally, the myocardial oxygen consumption required to offset the dissipation of the transmembrane proton gradient in the absence of ATP synthesis was not correlated with either MMR or SMR. The results support the idea that interindividual variation in aerobic metabolism, particularly MMR, is associated with variation in specific steps in the O2 transport cascade.
Subject(s)
Fundulidae , Animals , Cell Respiration , Energy Metabolism , Gills/anatomy & histology , Oxygen ConsumptionABSTRACT
Throughout the course of infection, many pathogens encounter bactericidal conditions that threaten the viability of the bacteria and impede the establishment of infection. Bile is one of the most innately bactericidal compounds present in humans, functioning to reduce the bacterial burden in the gastrointestinal tract while also aiding in digestion. It is becoming increasingly apparent that pathogens successfully resist the bactericidal conditions of bile, including bacteria that do not normally cause gastrointestinal infections. This review highlights the ability of Enterococcus, Staphylococcus, Klebsiella, Acinetobacter, Pseudomonas, Enterobacter (ESKAPE), and other enteric pathogens to resist bile and how these interactions can impact the sensitivity of bacteria to various antimicrobial agents. Given that pathogen exposure to bile is an essential component to gastrointestinal transit that cannot be avoided, understanding how bile resistance mechanisms align with antimicrobial resistance is vital to our ability to develop new, successful therapeutics in an age of widespread and increasing antimicrobial resistance.
Subject(s)
Anti-Bacterial Agents/metabolism , Bacteria/pathogenicity , Bile/metabolism , Drug Resistance, Bacterial , Anti-Bacterial Agents/pharmacology , Bacteria/classification , Bacteria/drug effects , Bacteria/metabolism , Biofilms/drug effects , Biofilms/growth & development , Humans , Intestine, Small/microbiology , Membrane Transport Proteins/genetics , Membrane Transport Proteins/metabolism , VirulenceABSTRACT
Standard metabolic rate (SMR), maximum metabolic rate (MMR), absolute aerobic scope (AAS) and critical oxygen tension (Pcrit) were determined for the Gulf killifish, Fundulus grandis, an ecologically dominant estuarine fish, acclimated to lowered salinity, elevated temperature and lowered oxygen concentration. Acclimation to low salinity resulted in a small, but significant, elevation of Pcrit (suggesting lower tolerance of hypoxia); acclimation to elevated temperature increased SMR, MMR, AAS and Pcrit; acclimation to low oxygen led to a small increase in SMR, but substantial decreases in MMR, AAS and Pcrit Variation in these metabolic traits among individuals was consistent and repeatable when measured during multiple control exposures over 7 months. Trait repeatability was unaffected by acclimation condition, suggesting that repeatability of these traits is not context dependent. There were significant phenotypic correlations between specific metabolic traits: SMR was positively correlated with MMR and Pcrit; MMR was positively correlated with AAS; and AAS was negatively correlated with Pcrit In general, within-individual variation contributed more than among-individual variation to these phenotypic correlations. The effects of acclimation on these traits demonstrate that aerobic metabolism is plastic and influenced by the conditions experienced by these fish in the dynamic habitats in which they occur; however, the repeatability of these traits and the correlations among them suggest that these traits change in ways that maintain the rank order of performance among individuals across a range of environmental variation.
Subject(s)
Basal Metabolism , Fundulidae , Oxygen Consumption , Acclimatization , Animals , Humans , OxygenABSTRACT
Excessive neutrophil infiltration of the lungs is a common contributor to immune-related pathology in many pulmonary disease states. In response to pathogenic infection, airway epithelial cells produce hepoxilin A3 (HXA3), initiating neutrophil transepithelial migration. Migrated neutrophils amplify this recruitment by producing a secondary gradient of leukotriene B4 (LTB4). We sought to determine whether this two-step eicosanoid chemoattractant mechanism could be exploited by the pathogen Pseudomonas aeruginosa. ExoU, a P. aeruginosa cytotoxin, exhibits phospholipase A2 (PLA2) activity in eukaryotic hosts, an enzyme critical for generation of certain eicosanoids. Using in vitro and in vivo models of neutrophil transepithelial migration, we evaluated the impact of ExoU expression on eicosanoid generation and function. We conclude that ExoU, by virtue of its PLA2 activity, augments and compensates for endogenous host neutrophil cPLA2α function, leading to enhanced transepithelial migration. This suggests that ExoU expression in P. aeruginosa can circumvent immune regulation at key signaling checkpoints in the neutrophil, resulting in exacerbated neutrophil recruitment.
Subject(s)
Bacterial Proteins/immunology , Leukotriene B4/immunology , Neutrophil Infiltration/immunology , Pseudomonas Infections/immunology , Transendothelial and Transepithelial Migration/immunology , Animals , Blotting, Western , Female , Humans , Mice , Mice, Inbred C57BL , Neutrophils/immunology , Pseudomonas aeruginosa/pathogenicity , Virulence/immunologyABSTRACT
BACKGROUND: Global concern about vitamin D deficiency has fuelled debates on photoprotection and the importance of solar exposure to meet vitamin D requirements. OBJECTIVES: To review the published evidence to reach a consensus on the influence of photoprotection by sunscreens on vitamin D status, considering other relevant factors. METHODS: An international panel of 13 experts in endocrinology, dermatology, photobiology, epidemiology and biological anthropology reviewed the literature prior to a 1-day meeting in June 2017, during which the evidence was discussed. Methods of assessment and determining factors of vitamin D status, and public health perspectives were examined and consequences of sun exposure and the effects of photoprotection were assessed. RESULTS: A serum level of ≥ 50 nmol L-1 25(OH)D is a target for all individuals. Broad-spectrum sunscreens that prevent erythema are unlikely to compromise vitamin D status in healthy populations. Vitamin D screening should be restricted to those at risk of hypovitaminosis, such as patients with photosensitivity disorders, who require rigorous photoprotection. Screening and supplementation are advised for this group. CONCLUSIONS: Sunscreen use for daily and recreational photoprotection does not compromise vitamin D synthesis, even when applied under optimal conditions. What's already known about this topic? Knowledge of the relationship between solar exposure behaviour, sunscreen use and vitamin D is important for public health but there is confusion about optimal vitamin D status and the safest way to achieve this. Practical recommendations on the potential impact of daily and/or recreational sunscreens on vitamin D status are lacking for healthy people. What does this study add? Judicious use of daily broad-spectrum sunscreens with high ultraviolet (UV) A protection will not compromise vitamin D status in healthy people. However, photoprotection strategies for patients with photosensitivity disorders that include high sun-protection factor sunscreens with high UVA protection, along with protective clothing and shade-seeking behaviour are likely to compromise vitamin D status. Screening for vitamin D status and supplementation are recommended in patients with photosensitivity disorders.
Subject(s)
Evidence-Based Medicine/standards , Skin Neoplasms/prevention & control , Sunlight/adverse effects , Sunscreening Agents/adverse effects , Vitamin D Deficiency/prevention & control , Vitamin D/blood , Consensus , Global Health/standards , Humans , Mass Screening/standards , Recreation , Reference Values , Skin/drug effects , Skin/metabolism , Skin/radiation effects , Skin Neoplasms/etiology , Sun Protection Factor , Sunscreening Agents/administration & dosage , Sunscreening Agents/chemistry , Ultraviolet Rays/adverse effects , Vitamin D/administration & dosage , Vitamin D/metabolism , Vitamin D Deficiency/blood , Vitamin D Deficiency/diagnosis , Vitamin D Deficiency/epidemiologyABSTRACT
The critical oxygen tension (Pcrit) for fishes is the oxygen level below which the rate of oxygen consumption (MO2 ) becomes dependent upon ambient oxygen partial pressure (PO2 ). We compare multiple curve-fitting approaches to estimate Pcrit of the Gulf killifish, Fundulus grandis, during closed and intermittent-flow respirometry. Fitting two line segments of MO2 versus PO2 produced high and variable estimates of Pcrit, as did nonlinear regression using a hyperbolic (Michaelis-Menten) function. Using nonlinear regression fit to an exponential (modified Weibull) function, or linear regression of MO2 versus PO2 at low PO2 , and determining Pcrit as the PO2 when MO2 equals standard metabolic rate (SMR) yielded values that were consistent across fish and among experimental trials. The magnitude of the difference in Pcrit determined by alternative calculation methods exceeded the differences determined in closed and intermittent-flow respirometry, highlighting the need to standardize analytical as well as experimental approaches in determining Pcrit.
Subject(s)
Fundulidae/physiology , Oxygen Consumption/physiology , Oxygen/blood , Animals , Basal Metabolism , Hypoxia , Models, Statistical , Oxygen/metabolism , Respiratory Physiological PhenomenaABSTRACT
Eicosanoids are a group of bioactive lipids that are shown to be important mediators of neutrophilic inflammation; selective targeting of their function confers therapeutic benefit in a number of diseases. Neutrophilic airway diseases, including cystic fibrosis, are characterized by excessive neutrophil infiltration into the airspace. Understanding the role of eicosanoids in this process may reveal novel therapeutic targets. The eicosanoid hepoxilin A3 is a pathogen-elicited epithelial-produced neutrophil chemoattractant that directs transepithelial migration in response to infection. Following hepoxilin A3-driven transepithelial migration, neutrophil chemotaxis is amplified through neutrophil production of a second eicosanoid, leukotriene B4 (LTB4). The rate-limiting step of eicosanoid generation is the liberation of arachidonic acid by phospholipase A2, and the cytosolic phospholipase A2 (cPLA2)α isoform has been specifically shown to direct LTB4 synthesis in certain contexts. Whether cPLA2α is directly responsible for neutrophil synthesis of LTB4 in the context of Pseudomonas aeruginosa-induced neutrophil transepithelial migration has not been explored. Human and mouse neutrophil-epithelial cocultures were used to evaluate the role of neutrophil-derived cPLA2α in infection-induced transepithelial signaling by pharmacological and genetic approaches. Primary human airway basal stem cell-derived epithelial cultures and micro-optical coherence tomography, a new imaging modality that captures two- and three-dimensional real-time dynamics of neutrophil transepithelial migration, were applied. Evidence from these studies suggests that cPLA2α expressed by neutrophils, but not epithelial cells, plays a significant role in infection-induced neutrophil transepithelial migration by mediating LTB4 synthesis during migration, which serves to amplify the magnitude of neutrophil recruitment in response to epithelial infection.
Subject(s)
Antigens, Human Platelet/metabolism , Cystic Fibrosis/immunology , Neutrophils/immunology , Pseudomonas Infections/immunology , Pseudomonas aeruginosa/immunology , Respiratory Mucosa/immunology , Transendothelial and Transepithelial Migration , 8,11,14-Eicosatrienoic Acid/analogs & derivatives , 8,11,14-Eicosatrienoic Acid/metabolism , Animals , Cell Communication , Cell Line , Chemotaxis , Coculture Techniques , Cytosol/metabolism , Humans , Leukotriene B4/metabolism , Mice , Neutrophils/microbiology , Respiratory Mucosa/microbiology , Respiratory Mucosa/pathology , Tomography, Optical CoherenceABSTRACT
The effects of passive integrated transponder (PIT) tagging on cortisol release, standard metabolic rate (SMR) and daily specific growth rate (GS ) were evaluated in the Gulf killifish, Fundulus grandis, a small estuarine fish native to the Gulf of Mexico. Cortisol release by individual fish was measured non-invasively prior to PIT tagging, immediately after tagging and once per week for 1 month following tagging. Within the first 2 h of tagging, cortisol release rates were significantly elevated compared with values measured prior to tagging and significantly higher than that of fish handled identically except not implanted with PIT tags. By 1 week after PIT tagging, cortisol release rates returned to control levels. SMR, determined by intermittent-flow respirometry and GS , defined as per cent change in body mass per day, were measured prior to PIT tagging and weekly for 1 month after tagging. Neither SMR nor GS was significantly different in tagged v. untagged fish for the duration of the study. One month after tagging, haematocrit, plasma cortisol, blood glucose and blood lactate did not differ between tagged and untagged individuals. Therefore, after a transient stress response that subsides within 1 week, PIT tagging had no significant effects on these physiological variables in F. grandis, validating its use as a method of marking this and other small fishes.
Subject(s)
Fundulidae/metabolism , Hydrocortisone/blood , Telemetry/adverse effects , Animals , Blood Glucose , Female , Fundulidae/growth & development , Gulf of Mexico , Hematocrit , Lactic Acid/blood , Male , Telemetry/instrumentationABSTRACT
Many fish experience daily cycles of hypoxia in the wild, but the physiological strategies for coping with intermittent hypoxia are poorly understood. We examined how killifish adjust O2 supply and demand during acute hypoxia, and how these responses are altered after prolonged acclimation to constant or intermittent patterns of hypoxia exposure. We acclimated killifish to normoxia (â¼20â kPaâ O2), constant hypoxia (2â kPa) or intermittent cycles of nocturnal hypoxia (12â h:12â h normoxia:hypoxia) for 28â days, and then compared whole-animal O2 consumption rates (MO2 ) and tissue metabolites during exposure to 12â h of hypoxia followed by reoxygenation in normoxia. Normoxia-acclimated fish experienced a pronounced 27% drop in MO2 during acute hypoxia, and modestly increased MO2 upon reoxygenation. They strongly recruited anaerobic metabolism during acute hypoxia, indicated by lactate accumulation in plasma, muscle, liver, brain, heart and digestive tract, as well as a transient drop in intracellular pH, and they increased hypoxia inducible factor (HIF)-1α protein abundance in muscle. Glycogen, glucose and glucose-6-phosphate levels suggested that glycogen supported brain metabolism in hypoxia, while the muscle used circulating glucose. Acclimation to constant hypoxia caused a stable â¼50% decrease in MO2 that persisted after reoxygenation, with minimal recruitment of anaerobic metabolism, suggestive of metabolic depression. By contrast, fish acclimated to intermittent hypoxia maintained sufficient O2 transport to support normoxic MO2 , modestly recruited lactate metabolism and increased MO2 dramatically upon reoxygenation. Both groups of hypoxia-acclimated fish had similar glycogen, ATP, intracellular pH and HIF-1α levels as normoxic controls. We conclude that different patterns of hypoxia exposure favour distinct strategies for matching O2 supply and O2 demand.
Subject(s)
Acclimatization/physiology , Fundulidae/metabolism , Hypoxia/physiopathology , Oxygen Consumption/physiology , Anaerobiosis/physiology , Animals , Glucose/metabolism , Glycogen/metabolism , Hydrogen-Ion Concentration , Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , Lactic Acid/metabolismABSTRACT
BACKGROUND: This study seeks to compare the performance of HRP2 (First Response) and pLDH/HRP2 (Combo) RDTs for falciparum malaria against microscopy and PCR in acutely ill febrile children at presentation and follow-up. METHODS: This is an interventional study that recruited children < 5 years who reported to health facilities with a history of fever within the past 72 h or a documented axillary temperature of 37.5 °C. Using a longitudinal approach, recruitment and follow-up of participants was done between January and May 2012. Based on results of HRP2-RDT screening, the children were grouped into one of the following three categories: (1) tested positive for malaria using RDT and received anti-malarial treatment (group 1, n = 85); (2) tested negative for malaria using RDT and were given anti-malarial treatment by the admitting physician (group 2, n = 74); or, (3) tested negative for malaria using RDT and did not receive any anti-malarial treatment (group 3, n = 101). Independent microscopy, PCR and Combo-RDT tests were done for each sample on day 0 and all follow-up days. RESULTS: Mean age of the study participants was 22 months and females accounted for nearly 50%. At the time of diagnosis, the mean body temperature was 37.9 °C (range 35-40.1 °C). Microscopic parasite density ranged between 300 and 99,500 parasites/µL. With microscopy as gold standard, the sensitivity of HRP2 and Combo-RDTs were 95.1 and 96.3%, respectively. The sensitivities, specificities and predictive values for RDTs were relatively higher in microscopy-defined malaria cases than in PCR positive-defined cases. On day 0, participants who initially tested negative for HRP2 were positive by microscopy (n = 2), Combo (n = 1) and PCR (n = 17). On days 1 and 2, five of the children in this group (initially HRP2-negative) tested positive by PCR alone. On day 28, four patients who were originally HRP2-negative tested positive for microscopy (n = 2), Combo (n = 2) and PCR (n = 4). CONCLUSION: The HRP2/pLDH RDTs showed comparable diagnostic accuracy in children presenting with an acute febrile illness to health facilities in a hard-to-reach rural area in Ghana. Nevertheless, discordant results recorded on day 0 and follow-up visits using the recommended RDTs means improved malaria diagnostic capability in malaria-endemic regions is necessary.
Subject(s)
Antimalarials/therapeutic use , Diagnostic Tests, Routine/statistics & numerical data , Fever/diagnosis , Malaria, Falciparum/diagnosis , Plasmodium falciparum/isolation & purification , Child, Preschool , Female , Fever/drug therapy , Fever/parasitology , Ghana , Humans , Infant , Infant, Newborn , Longitudinal Studies , Malaria, Falciparum/drug therapy , Malaria, Falciparum/parasitology , Male , Microscopy/methods , Polymerase Chain Reaction/methods , Sensitivity and SpecificityABSTRACT
OBJECTIVE: Oxidative stress and low-grade chronic inflammation stand out as key features of physiological skin ageing. The aim of this study was to examine in normal human epidermal keratinocytes (NHEK) and human dermal fibroblasts (HDF) grown in vitro, the antioxidant and anti-inflammatory properties of crocin, a carotenoid glycoside responsible for the colour of saffron. Moreover, considering the newly emerging field of skin glycobiology and the presence of two gentiobiosyl moieties in crocin, the effect of crocin on NHEK glycosylation pathways was for the first time investigated. METHODS: The anti-inflammatory and antioxidant activities of crocin were evaluated by in vitro assays of antioxidation activities, ELISA and microarray analysis. The effect of crocin on keratinocyte glycobiology was evaluated by proprietary GLYcoDiag lectin technologies and microarray analysis. RESULTS: Crocin is endowed with antioxidant potential against reactive oxygen species, protects squalene against UVA-induced peroxidation and prevents the release of inflammatory mediators. The expression of NF-kB-related genes and glycosylation-related genes is modulated in the presence of crocin. CONCLUSION: Results could designate this molecule as a promising skin ageing prevention cosmetic agent. Of note, some of these effects could be mediated by protein O-glycosylation and interaction of crocin with osidic receptors of keratinocytes.
Subject(s)
Carotenoids/pharmacology , Skin Aging/drug effects , Cell Membrane/metabolism , Cells, Cultured , Enzyme-Linked Immunosorbent Assay , Fibroblasts/cytology , Fibroblasts/metabolism , Glycosylation , Humans , Inflammation Mediators/metabolism , Keratinocytes/cytology , Keratinocytes/metabolism , Oligonucleotide Array Sequence Analysis , Oxidative Stress/drug effects , Polysaccharides/metabolism , Reactive Oxygen Species/metabolism , Skin/cytology , Skin/drug effects , Skin/metabolismABSTRACT
OBJECTIVE: To study the effects of the very high minerality Vichy Thermal Spring Water (VTSW) on human keratinocytes grown in vitro. METHODS: The effect of VTSW was monitored by full genome transcriptomic technology and immunofluorescence microscopy. RESULTS: In the presence of 50% VTSW, the expression of a number of skin homoeostasis-related genes is increased, specifically with respect to dermal-epidermal junction, epidermal cohesion and communication, keratinocyte proliferation-differentiation balance, antioxidant mechanisms and DNA repair. CONCLUSION: This work suggests that VTSW could be considered as an ingredient of potential interest to address some of the deleterious effects of skin ageing exposome.
Subject(s)
Cosmetics , Skin Aging , Water , Antioxidants/metabolism , Cell Differentiation , Cell Proliferation , DNA Damage , DNA Repair , Homeostasis , Humans , In Vitro Techniques , Keratinocytes , Microscopy, Fluorescence , Oxidative StressABSTRACT
The hypoxia-inducible factor (HIF) family of transcription factors plays central roles in the development, physiology, pathology, and environmental adaptation of animals. Because many aquatic habitats are characterized by episodes of low dissolved oxygen, fish represent ideal models to study the roles of HIF in the response to aquatic hypoxia. The estuarine fish Fundulus heteroclitus is found in habitats prone to hypoxia. It responds to low oxygen via behavioral, physiological, and molecular changes, and one member of the HIF family, HIF2α, has been previously described. Herein, cDNA sequencing, phylogenetic analyses, and genomic approaches were used to determine other members of the HIFα family from F. heteroclitus and their relationships to HIFα subunits from other vertebrates. In vitro and cellular approaches demonstrated that full-length forms of HIF1α, HIF2α, and HIF3α independently formed complexes with the ß-subunit, aryl hydrocarbon receptor nuclear translocator, to bind to hypoxia response elements and activate reporter gene expression. Quantitative PCR showed that HIFα mRNA abundance varied among organs of normoxic fish in an isoform-specific fashion. Analysis of the F. heteroclitus genome revealed a locus encoding a second HIF2α-HIF2αb-a predicted protein lacking oxygen sensing and transactivation domains. Finally, sequence analyses demonstrated polymorphism in the coding sequence of each F. heteroclitus HIFα subunit, suggesting that genetic variation in these transcription factors may play a role in the variation in hypoxia responses among individuals or populations.