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Ion channels are potentially exploitable as pharmacological targets to treat asthma. This study evaluated the role of KCa3.1 channels, encoded by Kcnn4, in regulating the gene expression of mouse airway epithelium and the development of asthma traits. We used the ovalbumin (OVA) challenge as an asthma model in wild-type and Kcnn4-/- mice, performed histological analysis, and measured serum IgE to evaluate asthma traits. We analyzed gene expression of isolated epithelial cells of trachea or bronchi using mRNA sequencing and gene ontology and performed Ussing chamber experiments in mouse trachea to evaluate anion secretion. Gene expression of epithelial cells from mouse airways differed between trachea and bronchi, indicating regional differences in the inflammatory and transepithelial transport properties of proximal and distal airways. We found that Kcnn4 silencing reduced mast cell numbers, mucus, and collagen in the airways, and reduced the amount of epithelial anion secretion in the OVA-challenged animals. In addition, gene expression was differentially modified in the trachea and bronchi, with Kcnn4 genetic silencing significantly altering the expression of genes involved in the TNF pathway, supporting the potential of KCa3.1 as a therapeutic target for asthma.
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Asma , Traqueia , Animais , Asma/genética , Asma/metabolismo , Asma/patologia , Brônquios/metabolismo , Modelos Animais de Doenças , Expressão Gênica , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/metabolismo , Traqueia/metabolismo , Traqueia/patologiaRESUMO
In mammals, the daily variation in the ecology of the intestinal microbiota is tightly coupled to the circadian rhythm of the host. On the other hand, a close correlation between increased body weight and light pollution at night has been reported in humans and animal models. However, the mechanisms underlying such weight gain in response to light contamination at night remain elusive. In the present study, we tested the hypothesis that dim light pollution at night alters the colonic microbiota of mice, which could correlate with weight gain in the animals. By developing an experimental protocol using a mouse model that mimics light contamination at night in urban residences (dLAN, dim light at night), we found that mice exposed to dLAN showed a significant weight gain compared with mice exposed to control standard light/dark (LD) photoperiod. To identify possible changes in the microbiota, we sampled two stages from the resting period of the circadian cycle of mice (ZT0 and ZT10) and evaluated them by high-throughput sequencing technology. Our results indicated that microbial diversity significantly differed between ZT0 and ZT10 in both LD and dLAN samples and that dLAN treatment impacted the taxonomic composition, functions, and interactions of mouse colonic microbiota. Together, these results show that bacterial taxa and microbial metabolic pathways might be involved with the mechanisms underlying weight gain in mice subjected to light contamination at night.
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Colo/microbiologia , Microbioma Gastrointestinal , Poluição Luminosa/efeitos adversos , Aumento de Peso , Animais , CamundongosRESUMO
AIMS: Links between resistin, insulin resistance (IR), and resistin-stimulated cytokine signaling remain unknown in Mexican-Americans. A Mexican-American cohort was examined to determine (1) relationships between circulating resistin and IR, (2) resistin's associations with cytokines and demographic and anthropometric variables, and (3) similar measurements with other adipokines. METHODS: For cross sectional analyses, 953 adults (367 males and 586 females) in the Cameron County Hispanic Cohort (CCHC) were stratified into three groups: normal glucose tolerance, prediabetes, and diabetes mellitus. Differences in resistin and other adipokine levels were examined using linear regression via unadjusted model (Model 1), model adjusted for cytokines (Model 2), and model further adjusted for demographic and anthropometric variables (Model 3). RESULTS: HOMA-IR increased with worsening glucose tolerance (p < 0.0001). In all models, resistin significantly decreased as glucose tolerance deteriorated. Model 3 resistin was positively associated with IL-1ß (p = 0.0252) and IL-8 (p < 0.0001), inversely associated with TNF-α (p = 0.0352), but nonsignificantly associated with IL-6 (p = 0.8671). Model 3 leptin was significantly lower in diabetes mellitus compared to other groups (p < 0.005) and positively associated with female sex (p < 0.0001), age (p = 0.024), and BMI (p < 0.0001), without significant cytokine associations. Adiponectin displayed no significant associations with glucose tolerance, but was significantly associated with sex, BMI, and lipids (Model 3). CONCLUSIONS: Resistin unexpectedly decreased as IR increased while supporting evidence of a resistin-stimulated cytokine pathway in this Mexican-American cohort. Leptin fell with elevated IR after adjusting for cytokines, demographic and anthropometric variables. Adiponectin nonsignificantly decreased as IR increased while showing significant associations with sex, BMI, and lipids.
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Resistência à Insulina , Americanos Mexicanos , Resistina/sangue , Adipocinas/sangue , Adulto , Estudos de Coortes , Feminino , Teste de Tolerância a Glucose , Humanos , Leptina/sangue , Modelos Lineares , Masculino , Análise MultivariadaRESUMO
We provide new evidence on the effect of adolescent health behaviors/outcomes (obesity, depression, smoking, and attention deficit hyperactivity disorder [ADHD]) on schooling attainment using the National Longitudinal Study of Adolescent to Adult Health. We take two different approaches to deal with omitted variable bias and reverse causality. Our first approach attends to the issue of reverse causality by estimating the effect of health polygenic scores (PGSs) on schooling. Second, we estimate the effect of adolescent health using sibling fixed-effects models that control for unmeasured genetic and family factors shared by siblings. We use the PGSs as additional controls in the sibling fixed-effects models to reduce concerns about residual confounding from sibling-specific genetic differences. We find consistent evidence across both approaches that being genetically predisposed to smoking and smoking regularly in adolescence reduces schooling attainment. Estimates for depression are more imprecise, but also suggest that a high genetic risk of depression and adolescent depression reduce schooling attainment. We find mixed evidence for ADHD. Our estimates suggest that having a high genetic risk for ADHD reduces grades of schooling, but we do not find any statistically significant negative effects of ADHD. Finally, we find no consistent evidence for a detrimental effect of obesity on schooling attainment.
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Saúde do Adolescente , Transtorno do Deficit de Atenção com Hiperatividade , Adolescente , Adulto , Transtorno do Deficit de Atenção com Hiperatividade/epidemiologia , Transtorno do Deficit de Atenção com Hiperatividade/genética , Escolaridade , Humanos , Estudos Longitudinais , Herança MultifatorialRESUMO
Monocarboxylate transporter 4 (MCT4) is an H+-coupled symporter highly expressed in metastatic tumors and at inflammatory sites undergoing hypoxia or the Warburg effect. At these sites, extracellular lactate contributes to malignancy and immune response evasion. Intriguingly, at 30-40 mm, the reported Km of MCT4 for lactate is more than 1 order of magnitude higher than physiological or even pathological lactate levels. MCT4 is not thought to transport pyruvate. Here we have characterized cell lactate and pyruvate dynamics using the FRET sensors Laconic and Pyronic. Dominant MCT4 permeability was demonstrated in various cell types by pharmacological means and by CRISPR/Cas9-mediated deletion. Respective Km values for lactate uptake were 1.7, 1.2, and 0.7 mm in MDA-MB-231 cells, macrophages, and HEK293 cells expressing recombinant MCT4. In MDA-MB-231 cells MCT4 exhibited a Km for pyruvate of 4.2 mm, as opposed to >150 mm reported previously. Parallel assays with the pH-sensitive dye 2',7'-bis-(carboxyethyl)-5-(and-6)-carboxyfluorescein (BCECF) indicated that previous Km estimates based on substrate-induced acidification were severely biased by confounding pH-regulatory mechanisms. Numerical simulation using revised kinetic parameters revealed that MCT4, but not the related transporters MCT1 and MCT2, endows cells with the ability to export lactate in high-lactate microenvironments. In conclusion, MCT4 is a high-affinity lactate transporter with physiologically relevant affinity for pyruvate.
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Ácido Láctico/metabolismo , Transportadores de Ácidos Monocarboxílicos/metabolismo , Proteínas Musculares/metabolismo , Transporte Biológico/efeitos dos fármacos , Sistemas CRISPR-Cas/genética , Linhagem Celular Tumoral , Diclofenaco/farmacologia , Fluoresceínas/química , Edição de Genes , Células HEK293 , Humanos , Concentração de Íons de Hidrogênio , Cinética , Macrófagos/citologia , Macrófagos/metabolismo , Transportadores de Ácidos Monocarboxílicos/antagonistas & inibidores , Transportadores de Ácidos Monocarboxílicos/genética , Proteínas Musculares/antagonistas & inibidores , Proteínas Musculares/genética , Isoformas de Proteínas/antagonistas & inibidores , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Ácido Pirúvico/metabolismoRESUMO
Anthocyanins are pigments with antihyperglycemic properties, and they are potential candidates for developing functional foods for the therapy or prevention of Diabetes mellitus type 2 (DM2). The mechanism of these beneficial effects of anthocyanins are, however, hard to explain, given their very low bioavailability due to poor intestinal absorption. We propose that free fatty acid receptor 1 (FFA1, also named GPR40), is involved in an inhibitory effect of the anthocyanidin delphinidin over intestinal glucose absorption. We show the direct effects of delphinidin on the intestine using jejunum samples from RF/J mice, and the human intestinal cell lines HT-29, Caco-2, and NCM460. By the use of specific pharmacological antagonists, we determined that delphinidin inhibits glucose absorption in both mouse jejunum and a human enterocytic cell line in a FFA1-dependent manner. Delphinidin also affects the function of sodium-glucose cotransporter 1 (SGLT1). Intracellular signaling after FFA1 activation involved cAMP increase and cytosolic Ca2+ oscillations originated from intracellular Ca2+ stores and were followed by store-operated Ca2+ entry. Taken together, our results suggest a new GPR-40 mediated local mechanism of action for delphinidin over intestinal cells that may in part explain its antidiabetic effect. These findings are promising for the search for new prevention and pharmacological treatment strategies for DM2 management.
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Antocianinas/farmacologia , Glucose/metabolismo , Intestinos/química , Jejuno/química , Receptores Acoplados a Proteínas G/metabolismo , Animais , Células CACO-2 , Cálcio/metabolismo , Linhagem Celular , Regulação da Expressão Gênica/efeitos dos fármacos , Células HT29 , Humanos , Intestinos/efeitos dos fármacos , Jejuno/efeitos dos fármacos , Camundongos , Transdução de Sinais/efeitos dos fármacosRESUMO
BACKGROUND & AIMS: The fluid secretion model predicts that intestinal obstruction disorders can be alleviated by promoting epithelial Cl(-) secretion. The adenosine 3',5'-cyclic monophosphate (cAMP)-activated anion channel CFTR mediates Cl(-)-dependent fluid secretion in the intestine. Although the role of the ClC-2 channel has not been determined in the intestine, this voltage-gated Cl(-) channel might compensate for the secretory defects observed in patients with cystic fibrosis and other chronic constipation disorders. We investigated whether mice that lack ClC-2 channels (Clcn2(-/-)) have defects in intestinal ion transport. METHODS: Immunolocalization and immunoblot analyses were used to determine the cellular localization and the amount of ClC-2 expressed in mouse early distal colon (EDC) and late distal colon (LDC). Colon sheets from wild-type and Clcn2(-/-) littermates were mounted in Ussing chambers to determine transepithelial bioelectrical parameters and Na(+), K(+), and Cl(-) fluxes. RESULTS: Expression of ClC-2 was higher in the basolateral membrane of surface cells in the EDC compared with the LDC, with little expression in crypts. Neither cAMP nor Ca(2+)-induced secretion of Cl(-) was affected in the EDC or LDC of Clcn2(-/-) mice, whereas the amiloride-sensitive short-circuit current was increased approximately 3-fold in Clcn2(-/-) EDC compared with control littermates. Conversely, electroneutral Na(+), K(+), and Cl(-) absorption was dramatically reduced in colons of Clcn2(-/-) mice. CONCLUSIONS: Basolateral ClC-2 channels are required for colonic electroneutral absorption of NaCl and KCl. The increase in the amiloride-sensitive short-circuit current in Clcn2(-/-) mice revealed a compensatory mechanism that is activated in the colons of mice that lack the ClC-2 channel.
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Canais de Cloreto/metabolismo , Colo/metabolismo , Absorção Intestinal , Mucosa Intestinal/metabolismo , Animais , Western Blotting , Canais de Cloro CLC-2 , Cloro/metabolismo , Constipação Intestinal/metabolismo , Feminino , Transporte de Íons , Masculino , Camundongos , Camundongos Knockout , Reação em Cadeia da Polimerase , Cloreto de Potássio/metabolismo , Cloreto de Sódio/metabolismoRESUMO
Although amphibian consumption by humans has been reported globally, this practice is not well studied despite its direct implications to the decline of amphibian populations. The International Union for Conservation of Nature (IUCN) recognizes the need to document the use and trade of species to be considered in assessing their extinction risk. Here the consumption of Duellmanohylaignicolor tadpoles is documented. It is a micro endemic species categorized as Near Threatened (NT) consumed in a traditional dish called "caldo de piedra" (stone soup) prepared by the Chinantec people (Tsa Ju Jmí') in Oaxaca, Mexico. Through conversations with local people and stream monitoring, the behavior of tadpoles of this species was documented and aspects of their exploitation and habitat use described. Places where caldo de piedra is still consumed were determined and using a spatial analysis with Geographic Information Systems, the distribution of the species in relation to those localities was analyzed. A number of other areas where tadpoles of this species might also occur and be exploited is predicted. In conclusion, the school behaviour, surface feeding, and the preference for deeper waterbodies that these tadpoles exhibit makes them vulnerable to being caught in large quantities. As they are consumed locally, are not commercialized, and the species distribution range is wider than caldo de piedra consumption, this implies a low risk for their populations. However, the tadpoles' reliance on streams with depths xÌ = 60 cm and flux xÌ = 0.65 m/s reduces the availability of sites for their optimal development.
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Bicarbonate secretion is a fundamental process involved in maintaining acid-base homeostasis. Disruption of bicarbonate entry into airway lumen, as has been observed in cystic fibrosis, produces several defects in lung function due to thick mucus accumulation. Bicarbonate is critical for correct mucin deployment and there is increasing interest in understanding its role in airway physiology, particularly in the initiation of lung disease in children affected by cystic fibrosis, in the absence of detectable bacterial infection. The current model of anion secretion in mammalian airways consists of CFTR and TMEM16A as apical anion exit channels, with limited capacity for bicarbonate transport compared to chloride. However, both channels can couple to SLC26A4 anion exchanger to maximise bicarbonate secretion. Nevertheless, current models lack any details about the identity of the basolateral protein(s) responsible for bicarbonate uptake into airway epithelial cells. We report herein that the electrogenic, sodium-dependent, bicarbonate cotransporter, SLC4A4, is expressed in the basolateral membrane of human and mouse airways, and that it's pharmacological inhibition or genetic silencing reduces bicarbonate secretion. In fully differentiated primary human airway cells cultures, SLC4A4 inhibition induced an acidification of the airways surface liquid and markedly reduced the capacity of cells to recover from an acid load. Studies in the Slc4a4-null mice revealed a previously unreported lung phenotype, characterized by mucus accumulation and reduced mucociliary clearance. Collectively, our results demonstrate that the reduction of SLC4A4 function induced a CF-like phenotype, even when chloride secretion remained intact, highlighting the important role SLC4A4 plays in bicarbonate secretion and mammalian airway function.
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Proteína 1 de Troca de Ânion do Eritrócito/metabolismo , Fibrose Cística , Animais , Bicarbonatos/metabolismo , Cloretos/metabolismo , Regulador de Condutância Transmembrana em Fibrose Cística/genética , Mamíferos/metabolismo , Camundongos , Fenótipo , Sódio/metabolismo , Simportadores de Sódio-Bicarbonato/genéticaRESUMO
ZnO-based dye-sensitized solar cells exhibit lower efficiencies than TiO2-based systems despite advantageous charge transport dynamics and versatility in terms of synthesis methods, which can be primarily ascribed to compatibility issues of ZnO with the dyes and the redox couples originally optimized for TiO2. We evaluate the performance of solar cells based on ZnO nanomaterial prepared by microwave-assisted solvothermal synthesis, using three fully organic benzothiadiazole-based dyes YKP-88, YKP-137, and MG-207, and alternative electrolyte solutions with the I-/I3 -, Co(bpy)3 2+/3+, and Cu(dmp)2 1+/2+ redox couples. The best cell performance is achieved for the dye-redox couple combination YKP-88 and Co(bpy)3 2+/3+, reaching an average efficiency of 4.7% and 5.0% for the best cell, compared to 3.7% and 3.9% for the I-/I3 - couple with the same dye. Electrical impedance spectroscopy highlights the influence of dye and redox couple chemistry on the balance of recombination and regeneration kinetics. Combined with the effects of the interaction of the redox couple with the ZnO surface, these aspects are shown to determine the solar cell performance. Minimodules based on the best systems in both parallel and series configurations reach 1.5% efficiency for an area of 23.8 cm2.
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The montane cloud forests of the Sierra Madre de Oaxaca (SMO) host a remarkable herpetofauna diversity and represent one of the most important areas of endemism for Mexico and Mesoamerica. Although the area has been previously studied, most of the extant records for this group are biased to locations accessed by paved roads. In addition, an important proportion of this territory is conserved by Indigenous and Community Conservation Areas (ICCA), but little information of the species occurring within these areas exists. Therefore, information on the distribution of many endemic taxa in this region to date is either underestimated or incomplete. With the aim of increasing the ecological and distributional knowledge of this group in remote areas, we carried out field surveys in Santa Cruz Tepetotutla Oaxaca, a locality 25 km in a straight line to the closest paved road that conserves 9,670 ha of land through the ICCAs modality. Surveys were made during 2018 and 2019, including both dry and wet seasons. A total of 40 species of amphibians and reptiles were recorded: 32.5% of these records represent distributional range extensions, while 20% represent altitudinal range extensions. A total of 17.5% are records of species under a high risk category, highlighting both the relevance of studying remote areas to increase species population knowledge and the role of community conservation actions for species persistence. Finally, our records include the rediscovery of Rhadinella schistosa, a species undetected for more than 50 years.
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Mice have proven to be powerful models for the study of human physiology and pathophysiology. With the advent of techniques for genomic manipulation, the possibilities for studying inherited diseases in this convenient laboratory mammal are increasing by the day. It has been reported that when knocking out or otherwise modifying genes of interest in mice, the phenotype obtained can vary markedly depending on the genetic background of the animals used in the study. The aim of this work was to study whether the genetic background can influence the characteristics of fluid and electrolyte transepithelial transport in the distal colon of three mouse strains most in use in our and other laboratories. Ussing chamber recordings revealed that the colons of C57Bl/6J, Sv 129 and Black Swiss animals have distinctive responses to the calcium agonists carbachol and histamine that are not explained by the presence of different types of muscarinic and histaminergic receptors in these tissues. We have also found differences in the cAMP-activated, KCNMA1-channel-dependent potassium secretion between the strains. We interpret this to indicate a unique distribution of KCNMA1 channels in lower parts of the crypt of Sv 129 colonic epithelium compared with that of C57Bl/6J and Black Swiss animals. The reported differences should be taken into account when choosing the genetic background of animals to be used for genetic modification.
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Colo/metabolismo , Eletrólitos/metabolismo , Mucosa Intestinal/metabolismo , Camundongos Endogâmicos/fisiologia , Potássio/metabolismo , 1-Metil-3-Isobutilxantina/farmacologia , Animais , Bário/farmacologia , Bumetanida/farmacologia , Carbacol/farmacologia , Colforsina/farmacologia , Colo/fisiologia , AMP Cíclico/farmacologia , Fezes/química , Histamina/farmacologia , Antagonistas dos Receptores Histamínicos/farmacologia , Mucosa Intestinal/fisiologia , Subunidades alfa do Canal de Potássio Ativado por Cálcio de Condutância Alta/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos/genética , Antagonistas Muscarínicos/farmacologia , Receptores Histamínicos H1/efeitos dos fármacos , Receptores Muscarínicos/efeitos dos fármacos , Simportadores de Cloreto de Sódio-Potássio/efeitos dos fármacos , Membro 2 da Família 12 de Carreador de Soluto , Água/análiseRESUMO
We estimate the effect of BMI on mental health for young adults and elderly individuals using data from the National Longitudinal Study of Adolescent Health and the Health & Retirement Study. To tackle confounding due to unobserved factors, we exploit variation in a polygenic score (PGS) for BMI within two related econometric methods that differ in the assumptions they employ. First, we use the BMI PGS as an IV and adjust for PGSs for other factors (depression and educational attainment) that may invalidate this IV. We find a large statistically significant effect of BMI on mental health for the elderly: a 5â¯kg/m2 increase in BMI (a difference equivalent to moving from overweight to obese) increases the probability of depression by 29 %. In contrast, for young adults the IV estimates are statistically and economically insignificant. We show that IV estimates likely have to be interpreted as identifying a weighted average of effects of BMI on mental health mostly for compliers on the upper quantiles of the BMI distribution. Second, we use the BMI PGS as an "imperfect" IV and estimate an upper bound on the average treatment effect for the population. The estimated upper bounds are consistent with the conclusions from the IV estimates.
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Índice de Massa Corporal , Depressão/epidemiologia , Depressão/genética , Obesidade/epidemiologia , Obesidade/genética , Adolescente , Fatores Etários , Idoso , Feminino , Marcadores Genéticos , Inquéritos Epidemiológicos , Humanos , Estudos Longitudinais , Masculino , Saúde Mental/estatística & dados numéricos , Modelos Econométricos , Sobrepeso/epidemiologia , Sobrepeso/genética , Fatores Sexuais , Fatores Socioeconômicos , Adulto JovemRESUMO
Lactate, the product of aerobic glycolysis, plays a dual role as fuel and intercellular signal in inflammation, immune evasion, and tumor progression. The production of lactate by macrophages has been associated with their polarization and function. Here we describe imaging protocols to characterize the metabolism of cultured human macrophages using a genetically encoded fluorescent sensor-specific for lactate. By superfusing cultures with increasing lactate concentrations and pharmacological inhibitors, it is possible to estimate the kinetic parameters of monocarboxylate transporter 4 (MCT4) and lactate production. Practical advice is given regarding sensor expression, imaging, and data analysis. The spatiotemporal resolution of this technique is amenable to the study of fast events at the single-cell level in different immune and other cell types.
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Ácido Láctico/metabolismo , Macrófagos/metabolismo , Transporte Biológico/fisiologia , Linhagem Celular , Corantes Fluorescentes/metabolismo , Humanos , Cinética , Transportadores de Ácidos Monocarboxílicos/metabolismo , Células THP-1/metabolismoRESUMO
Airway mucociliary clearance (MCC) is the main mechanism of lung defense keeping airways free of infection and mucus obstruction. Airway surface liquid volume, ciliary beating, and mucus are central for proper MCC and critically regulated by sodium absorption and anion secretion. Impaired MCC is a key feature of muco-obstructive diseases. The calcium-activated potassium channel KCa.3.1, encoded by Kcnn4, participates in ion secretion, and studies showed that its activation increases Na+ absorption in airway epithelia, suggesting that KCa3.1-induced hyperpolarization was sufficient to drive Na+ absorption. However, its role in airway epithelium is not fully understood. We aimed to elucidate the role of KCa3.1 in MCC using a genetically engineered mouse. KCa3.1 inhibition reduced Na+ absorption in mouse and human airway epithelium. Furthermore, the genetic deletion of Kcnn4 enhanced cilia beating frequency and MCC ex vivo and in vivo. Kcnn4 silencing in the Scnn1b-transgenic mouse (Scnn1btg/+), a model of muco-obstructive lung disease triggered by increased epithelial Na+ absorption, improved MCC, reduced Na+ absorption, and did not change the amount of mucus but did reduce mucus adhesion, neutrophil infiltration, and emphysema. Our data support that KCa3.1 inhibition attenuated muco-obstructive disease in the Scnn1btg/+ mice. K+ channel modulation may be a therapeutic strategy to treat muco-obstructive lung diseases.
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Canais de Potássio Ativados por Cálcio de Condutância Intermediária/genética , Pneumopatias Obstrutivas/etiologia , Depuração Mucociliar/fisiologia , Animais , Cálcio/metabolismo , Células Cultivadas , Cílios/efeitos dos fármacos , Cílios/metabolismo , Modelos Animais de Doenças , Epitélio/metabolismo , Feminino , Humanos , Canais de Potássio Ativados por Cálcio de Condutância Intermediária/antagonistas & inibidores , Canais de Potássio Ativados por Cálcio de Condutância Intermediária/metabolismo , Pulmão/fisiopatologia , Pneumopatias Obstrutivas/genética , Masculino , Camundongos Endogâmicos C57BL , Camundongos Mutantes , Camundongos Transgênicos , Depuração Mucociliar/efeitos dos fármacos , Sódio/metabolismoRESUMO
Nosema ceranae is an emerging pathogen of the western honey bee (Apis mellifera L.), and thus its seasonality and impact on bee colonies is not sufficiently documented for North America. This study was conducted to determine the infection intensity, prevalence, and viability of N. ceranae in >200 honey bee colonies during spring, summer, and fall, in a North American region. We also determined the relationship of N. ceranae infections with colony populations, food stores, bee survivorship, and overwinter colony mortality. The highest rates of N. ceranae infection, prevalence, and spore viability were found in the spring and summer, while the lowest were recorded in the fall. N. ceranae spore viability was significantly correlated with its prevalence and infection intensity in bees. Threshold to high levels of N. ceranae infections (>1,000,000 spores/bee) were significantly associated with reduced bee populations and food stores in colonies. Furthermore, worker bee survivorship was significantly reduced by N. ceranae infections, although no association between N. ceranae and winter colony mortality was found. It is concluded that N. ceranae infections are highest in spring and summer and may be detrimental to honey bee populations and colony productivity. Our results support the notion that treatment is justified when infections of N. ceranae exceed 1,000,000 spores/bee.
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[This corrects the article DOI: 10.3389/fphys.2019.00694.].
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Calcium-activated anion secretion is expected to ameliorate cystic fibrosis, a genetic disease that carries an anion secretory defect in exocrine tissues. Human patients and animal models of the disease that present a mild intestinal phenotype have been postulated to bear a compensatory calcium-activated anion secretion in the intestine. TMEM16A is calcium-activated anion channel whose presence in the intestinal epithelium is contradictory. We aim to test the functional expression of TMEM16A using animal models with Cftr and/or Tmem16a intestinal silencing. Expression of TMEM16A was studied in a wild type and intestinal Tmem16a knockout mice by mRNA-seq, mass-spectrometry, q-PCR, Western blotting and immunolocalization. Calcium-activated anion secretion was recorded in the ileum and proximal colon of these animals including intestinal Cftr knockout and double mutants with dual Tmem16a and Cftr intestinal ablation. Mucus homeostasis was studied by immune-analysis of Mucin-2 (Muc2) and survival curves were recorded. Tmem16a transcript was found in intestine. Nevertheless, protein was barely detected in colon samples. Electrophysiological measurements demonstrated that the intestinal deletion of Tmem16a did not change calcium-activated anion secretion induced by carbachol or ATP in ileum and proximal colon. Muc2 architecture was not altered by Tmem16a silencing as was observed when Cftr was deleted from mouse intestine. Tmem16a silencing neither affected animal survival nor modified the lethality observed in the intestinal Cftr-null mouse. Our results demonstrate that TMEM16A function in the murine intestine is not related to electrogenic calcium-activated anion transport and does not affect mucus homeostasis and survival of animals.
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Nearly 70% of cystic fibrosis (CF) patients bear the phenylalanine-508 deletion but disease severity differs greatly, and is not explained by the existence of different mutations in compound heterozygous. Studies demonstrated that genes other than CFTR relate to intestinal disease in humans and CF-mouse. Kcnn4, the gene encoding the calcium-activated potassium channel KCa3.1, important for intestinal secretion, is present in a locus linked with occurrence of intestinal CF-disease in mice and humans. We reasoned that it might be a CF-modifier gene and bred a CF-mouse with Kcnn4 silencing, finding that lethality was almost abolished. Silencing of Kcnn4 did not improve intestinal secretory functions, but rather corrected increased circulating TNF-α level and reduced intestinal mast cell increase. Given the importance of mast cells in intestinal disease additional double mutant CF-animals were tested, one lacking mast cells (C-kitW-sh/W-sh) and Stat6-/- to block IgE production. While mast cell depletion had no effect, silencing Stat6 significantly reduced lethality. Our results show that Kcnn4 is an intestinal CF modifier gene partially acting through a STAT6-dependent mechanism.