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PURPOSE: Extracellular acidification is a major component of tissue inflammation, including airway inflammation in asthmatics. However, its physiological/pathophysiological significance in bronchial function is not fully understood. Currently, the functional role of extracellular acidification on bronchial contraction was explored. METHODS: Left main bronchi were isolated from male BALB/c mice. Epithelium-removed tissues were exposed to acidic pH under submaximal contraction induced by 10-5 M acetylcholine in the presence or absence of a COX inhibitor indomethacin (10-6 M). Effects of AH6809 (10-6 M, an EP2 receptor antagonist), BW A868C (10-7 M, a DP receptor antagonist) and CAY10441 (3×10-6 M, an IP receptor antagonist) on the acidification-induced change in tension were determined. The release of prostaglandin E2 (PGE2) from epithelium-denuded tissues in response to acidic pH was assessed using an ELISA. RESULTS: In the bronchi stimulated with acetylcholine, change in the extracellular pH from 7.4 to 6.8 caused a transient augmentation of contraction followed by a sustained relaxing response. The latter inhibitory response was abolished by indomethacin and AH6809 but not by BW A868C or CAY10441. Both indomethacin and AH6809 significantly increased potency and efficacy of acetylcholine at pH 6.8. Stimulation with low pH caused an increase in PGE2 release from epithelium-denuded bronchi. Interestingly, the acidic pH-induced bronchial relaxation was significantly reduced in a murine asthma model that had a bronchial hyperresponsiveness to acetylcholine. CONCLUSION: Taken together, extracellular acidification could inhibit the bronchial contraction via autocrine activation of EP2 receptors. The diminished acidic pH-mediated inhibition of bronchial tone may contribute to excessive bronchoconstriction in inflamed airways such as asthma.
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Acetilcolina , Asma , Compostos de Benzil , Imidazóis , Animais , Masculino , Camundongos , Acetilcolina/farmacologia , Brônquios , Dinoprostona/metabolismo , Concentração de Íons de Hidrogênio , Indometacina/farmacologia , Inflamação , Contração Muscular , Camundongos Endogâmicos BALB CRESUMO
For powder compaction, the Kawakita equation has been used to estimate the powder behavior inside the die. The compression pressure exerted on powders is not homogeneous because of the friction on the die wall. However, the yield pressure and porosity estimated using the Kawakita equation are defined based on the assumption that homogeneous voids and compression pressure are distributed throughout the powder bed. In this study, an extended Kawakita equation was derived by considering the variation in the compression pressure as it corresponds to the distance from the loading punch surface. The yield time section estimated from the extended Kawakita equation was wider than that which was estimated via the classical equation. This result is consistent with the assumptions used to derive the extended Kawakita equation. Furthermore, a comparison of the porosity changes before and after the yield pressure was applied indicate that the direct cause of the yield is the spatial constraints of the powder particles. Equivalent stresses were defined to clarify the critical factor that constitutes the extended Kawakita equation. As a result, "taking into account the die wall friction" was considered to be the critical factor in the extended Kawakita equation. As these findings were theoretically determined by the extended Kawakita equation, a useful model was derived for a better understanding of powder compaction in die.
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Pós , Pressão , Porosidade , Comprimidos , Composição de MedicamentosRESUMO
PURPOSE: Extracellular acidification is a major component of tissue inflammation, including airway inflammation. The extracellular proton-sensing mechanisms are inherent in various cells including airway structural cells, although their physiological and pathophysiological roles in bronchial smooth muscles (BSMs) are not fully understood. In the present study, to explore the functional role of extracellular acidification on the BSM contraction, the isolated mouse BSMs were exposed to acidic pH under contractile stimulation. METHODS AND RESULTS: The RT-PCR analyses revealed that the proton-sensing G protein-coupled receptors were expressed both in mouse BSMs and cultured human BSM cells. In the mouse BSMs, change in the extracellular pH from 8.0 to 6.8 caused an augmentation of contraction induced by acetylcholine. Interestingly, the acidic pH-induced BSM hyper-contraction was further augmented in the mice that were sensitized and repeatedly challenged with ovalbumin antigen. In this animal model of asthma, upregulations of G protein-coupled receptor 68 (GPR68) and GPR65, that were believed to be coupled with Gq and Gs proteins respectively, were observed, indicating that the acidic pH could cause hyper-contraction probably via an activation of GPR68. However, psychosine, a putative antagonist for GPR68, failed to block the acidic pH-induced responses. CONCLUSION: These findings suggest that extracellular acidification contributes to the airway hyperresponsiveness, a characteristic feature of bronchial asthma. Further studies are required to identify the receptor(s) responsible for sensing extracellular protons in BSM cells.
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Asma , Hiper-Reatividade Brônquica , Acetilcolina/efeitos adversos , Acetilcolina/metabolismo , Animais , Brônquios , Hiper-Reatividade Brônquica/metabolismo , Humanos , Concentração de Íons de Hidrogênio , Inflamação/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Músculo Liso/metabolismo , Ovalbumina , Prótons , Psicosina/efeitos adversos , Psicosina/metabolismo , Receptores Acoplados a Proteínas G/genética , Receptores Acoplados a Proteínas G/metabolismoRESUMO
GCN1 is an evolutionarily-conserved ribosome-binding protein that mediates the amino acid starvation response as well as the ribotoxic stress response. We previously demonstrated that Gcn1 mutant mice lacking the GCN2-binding domain suffer from growth retardation and postnatal lethality via GCN2-independent mechanisms, while Gcn1-null mice die early in embryonic development. In this study, we explored the role of GCN1 in adult mice by generating tamoxifen-inducible conditional knockout (CKO) mice. Unexpectedly, the Gcn1 CKO mice showed body weight loss during tamoxifen treatment, which gradually recovered following its cessation. They also showed decreases in liver weight, hepatic glycogen and lipid contents, blood glucose and non-esterified fatty acids, and visceral white adipose tissue weight with no changes in food intake and viability. A decrease of serum VLDL suggested that hepatic lipid supply to the peripheral tissues was primarily impaired. Liver proteomic analysis revealed the downregulation of mitochondrial ß-oxidation that accompanied increases of peroxisomal ß-oxidation and aerobic glucose catabolism that maintain ATP levels. These findings show the involvement of GCN1 in hepatic lipid metabolism during tamoxifen treatment in adult mice.
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Proteínas de Saccharomyces cerevisiae , Animais , Lipídeos , Fígado/metabolismo , Glicogênio Hepático/metabolismo , Camundongos , Camundongos Knockout , Fatores de Alongamento de Peptídeos/metabolismo , Proteínas Serina-Treonina Quinases , Proteômica , Proteínas de Ligação a RNA/metabolismo , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Tamoxifeno/efeitos adversos , Tamoxifeno/metabolismo , Transativadores/metabolismo , Redução de PesoRESUMO
From the viewpoint of self-medication, it is valuable to develop patient-friendly scored tablets that possess dividing uniformity. In this context, we attempted to optimize the preparation conditions for a tablet with a unique shape, such as a concavely curved scored tablet (CCST). Employing a design of experiment and a response surface method incorporating a thin-plate spline interpolation, and a bootstrap resampling technique, the optimal preparation conditions for CCST were successfully developed. To make it possible to scaleup the optimal solution estimated on a trial-scale, a Bayesian estimation was applied. Credible ranges of critical responses in large-scale manufacturing were estimated as a posterior probability from the trial-scale experiment as a prior probability. In terms of the large-scale manufacturing, the possibility of solving the scaleup problem was suggested using Bayesian estimation. Furthermore, a simulation study using a finite element method revealed that strong tensile stresses generated along the tip of the score line in CCST when an outer force was applied to the back surface of CCST. An advantage in dividing uniformity is indicated by the unique shape of CCST.
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Comprimidos/química , Teorema de Bayes , Química Farmacêutica , Humanos , Modelos Químicos , Solubilidade , Propriedades de Superfície , Comprimidos/farmacologia , Resistência à TraçãoRESUMO
Progranulin (PGRN) is a multifunctional growth factor expressed in central nervous system. Although PGRN expression is regulated by various stressors, its precise role(s) and regulatory mechanism(s) remain elusive. In this study, we used HT22 cells to investigate the physiological implications of oxidative stress-induced PGRN expression and the regulation of PGRN expression by oxidative stress. We observed that p38 MAP kinase was activated upon the addition of H2O2, and a selective p38 MAP kinase inhibitor attenuated PGRN induction by H2O2. To explore the physiological role(s) of the PGRN induction, we first confirmed H2O2-dependent responses of HT22 cells and found that the length and number of neurites were increased by H2O2. Pgrn knockdown experiments suggested that these changes were mediated by H2O2-induced PGRN expression, at least in part. Overall, the results suggested that an increase in oxidative stress in HT22 cells induced PGRN expression via p38 MAP kinase pathway, thereby controlling neurite outgrowth.
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Peróxido de Hidrogênio , Progranulinas , Animais , Sistema de Sinalização das MAP Quinases , Crescimento NeuronalRESUMO
The importance of understanding the fate of nitrate (NO3-), which is the dominant N species transferred from terrestrial to aquatic ecosystems, has been increasing because global nitrogen loads have dramatically increased following industrialization. Dissimilatory nitrate reduction to ammonium (DNRA) and denitrification are both microbial processes that use NO3- for respiration. Compared to denitrification, quantitative determinations of the DNRA activity have been carried out only to a limited extent. This has led to an insufficient understanding of the importance of DNRA in NO3- transformations and the regulating factors of this process. The objective of this paper is to provide a detailed procedure for the measurement of the potential DNRA rate in environmental samples. In brief, the potential DNRA rate can be calculated from the 15N-labeled ammonium (15NH4+) accumulation rate in 15NO3- added incubation. The determination of the 14NH4+ and 15NH4+ concentrations described in this paper is comprised of the following steps. First, the NH4+ in the sample is extracted and trapped on an acidified glass filter as ammonium salt. Second, the trapped ammonium is eluted and oxidized to NO3- via persulfate oxidation. Third, the NO3- is converted to N2O via an N2O reductase deficient denitrifier. Finally, the converted N2O is analyzed using a previously developed quadrupole gas chromatography-mass spectrometry system. We applied this method to salt marsh sediments and calculated their potential DNRA rates, demonstrating that the proposed procedures allow a simple and more rapid determination compared to previously described methods.
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Compostos de Amônio/metabolismo , Nitratos/metabolismo , Isótopos de Nitrogênio/metabolismo , Óxido Nitroso/metabolismo , Calibragem , Sedimentos Geológicos/química , Nitritos/isolamento & purificação , Oxirredução , Oxigênio/isolamento & purificação , Politetrafluoretileno , Pseudomonas/metabolismo , Fatores de TempoRESUMO
The behavior of the structural orthorhombic-tetragonal phase transition of Pr2-xNdxNiO4+δ, a candidate material for solid oxide fuel cells and oxygen permeation membranes, was investigated by differential scanning calorimetry and thermogravimetry (TG), under controlled oxygen partial pressures, P(O2). The structural phase transition temperature, TP, of Pr2-xNdxNiO4+δ increased with increasing Nd content, x, or increasing P(O2). The phase transitions of all compositions involved discrete variations in the oxygen content, Δδ, which were observed in the TG curves under various P(O2) values. Δδ of Pr2-xNdxNiO4+δ with 0.5 ≤x≤ 1.5 were between those of Nd2NiO4+δ and Pr2NiO4+δ, regardless of P(O2), and were slightly increased with decreasing P(O2). It was proposed that the effect of the valence change of the Pr ion on Δδ was decreased with increasing Nd content. The standard enthalpy change, ΔH°, and entropy change, ΔS°, of the phase transition were estimated from the Ellingham diagrams and van't Hoff plots, which were prepared from the relationship between P(O2) and TP using an ideal solution model. ΔS° was decreased with increasing Nd content for the specimens with 0.0 ≤x≤ 1.5. The ΔH° of Pr2-xNdxNiO4+δ with 0.0 ≤x≤ 1.5 was almost constant for all Nd contents. The increase in the phase transition temperature of Pr2-xNdxNiO4+δ with increasing x from 0.0 to 1.5 was successfully explained using the calculated values of ΔH° and ΔS°.
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This study investigated the efficiency of intensive membrane cleaning for membrane bioreactors (MBRs) using a combination of mechanical scouring with granules and chemically enhanced backwashing (CEB). The implementation of such intensive cleaning was possible with ceramic flat-sheet membranes. Experiments were carried out using bench-scale MBRs at an existing wastewater treatment plant. First, CEB with NaClO was investigated in terms of the CEB frequency, duration, and concentration of the chemical reagent. CEB carried out for 60 min every 6 h, with 50 ppm of NaClO, was found to be effective, and it enabled an MBR to operate at 50 LMH, two to three times higher than the flux of full-scale MBRs. However, these CEB conditions were insufficient when the temperature was low (i.e. in winter), when an adhesive gel layer formed on the membrane surface. Its high resistance to cleaning might be explained by the increased levels of soluble microbial products and/or the presence of algal cells. Alkaline-assisted CEB, with NaClO (pH 12) and an increase in the volume of granules in the membrane tank, solved this problem. With the modified cleaning method, the fouling could be almost perfectly controlled at low-temperature conditions, such as 13 °C. MBRs may be regarded as fouling-free MBRs when the proposed cleaning method is used with ceramic flat-sheet membranes. Most real-world MBR operations operate with lower fluxes than the flux examined in this study, and at higher temperatures.
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Eliminação de Resíduos Líquidos , Águas Residuárias , Reatores Biológicos , Cerâmica , Membranas Artificiais , TemperaturaRESUMO
Amino acids exert many biological functions, serving as allosteric regulators and neurotransmitters, as constituents in proteins and as nutrients. GCN2-mediated phosphorylation of eukaryotic initiation factor 2 alpha (elF2α) restores homeostasis in response to amino acid starvation (AAS) through the inhibition of the general translation and upregulation of amino acid biosynthetic enzymes and transporters by activating the translation of Gcn4 and ATF4 in yeast and mammals, respectively. GCN1 is a GCN2-binding protein that possesses an RWD binding domain (RWDBD) in its C-terminus. In yeast, Gcn1 is essential for Gcn2 activation by AAS; however, the roles of GCN1 in mammals need to be established. Here, we revealed a novel role of GCN1 that does not depend on AAS by generating two Gcn1 mutant mouse lines: Gcn1-knockout mice (Gcn1 KO mice (Gcn1-/-)) and RWDBD-deleted mutant mice (Gcn1ΔRWDBD mice). Both mutant mice showed growth retardation, which was not observed in the Gcn2 KO mice, such that the Gcn1 KO mice died at the intermediate stage of embryonic development because of severe growth retardation, while the Gcn1ΔRWDBD embryos showed mild growth retardation and died soon after birth, most likely due to respiratory failure. Extension of pregnancy by 24 h through the administration of progesterone to the pregnant mothers rescued the expression of differentiation markers in the lungs and prevented lethality of the Gcn1ΔRWDBD pups, indicating that perinatal lethality of the Gcn1ΔRWDBD embryos was due to simple growth retardation. Similar to the yeast Gcn2/Gcn1 system, AAS- or UV irradiation-induced elF2α phosphorylation was diminished in the Gcn1ΔRWDBD mouse embryonic fibroblasts (MEFs), suggesting that GCN1 RWDBD is responsible for GCN2 activity. In addition, we found reduced cell proliferation and G2/M arrest accompanying a decrease in Cdk1 and Cyclin B1 in the Gcn1ΔRWDBD MEFs. Our results demonstrated, for the first time, that GCN1 is essential for both GCN2-dependent stress response and GCN2-independent cell cycle regulation.
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Ciclo Celular , Proliferação de Células , Desenvolvimento Fetal , Proteínas de Ligação a RNA/metabolismo , Estresse Fisiológico , Transativadores/metabolismo , Animais , Proteína Quinase CDC2/metabolismo , Células Cultivadas , Ciclina B1/metabolismo , Fibroblastos/metabolismo , Células HeLa , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas de Ligação a RNA/genética , Transativadores/genéticaRESUMO
Polychromium-oxo-deposited TiO2 (CrIII x Oy /TiO2 ) electrodes were fabricated by a simple electrochemical technique by using different TiO2 basal electrodes (anatase, rutile, and mixed polymorphic phases P25) as earth-abundant photoanodes for visible-light-driven water oxidation. The high-resolution transmission electron microscopy (HR-TEM) observation illustrated that an CrIII x Oy layer with approximately 2-3â nm thickness was formed on the surface of the crystalline TiO2 particles. Upon visible-light irradiation of the electrodes, the photoanodic current based on water oxidation was generated at the CrIII x Oy /TiO2 electrodes. However, the wavelength (below 620â nm) for photocurrent generation at CrIII x Oy /TiO2 -rutile was longer than that (below 560â nm) at CrIII x Oy /TiO2 -P25 by 60â nm, which is in agreement with the difference (0.2â eV) in the conduction band (CB) edge energy between rutile and anatase TiO2 . This gives a quantitative account for the photocurrent generation based on interfacial charge transfer (IFCT) from Cr 3d of the deposited CrIII x Oy layer to the TiO2 CB. The photocurrent generated for CrIII x Oy /TiO2 -rutile was higher than that for CrIII x Oy /TiO2 -anatase, which is ascribed to 1)â more effective CrIII x Oy deposition on the rutile particles, 2)â a larger electrolyte/CrIII x Oy interface for water oxidation as a result of smaller rutile particles (ca. 30-40â nm) compared with larger P25 particles (ca. 40-80â nm), and 3)â more effective use of visible light owing to the low energy IFCT transition of rutile.
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This study is the first to report on the detection of biogenic amines in an aqueous solution using an organic field-effect transistor (OFET) device with an extended gate electrode modified with a layer of diamine oxidase and a horseradish peroxidase osmium-redox polymer. The limit of detection (LOD) for histamine was estimated to be 1.2 µM. These results reveal that extended-gate type OFET devices are highly suitable enzyme-based biosensors for the detection of biogenic amine levels.
Assuntos
Aminas Biogênicas/análise , Técnicas Biossensoriais/instrumentação , Transistores Eletrônicos , Água/química , Amina Oxidase (contendo Cobre)/química , Amina Oxidase (contendo Cobre)/metabolismo , Aminas Biogênicas/química , Eletrodos , Enzimas Imobilizadas/química , Enzimas Imobilizadas/metabolismo , Histamina/análise , Histamina/química , Peroxidase do Rábano Silvestre/química , Peroxidase do Rábano Silvestre/metabolismo , Limite de Detecção , Oxirredução , SoluçõesRESUMO
Biosensors based on organic field effect transistors (OFETs) are one of the more promising device applications in organic electronics. However, OFET-based biosensors are still in their early stages of development compared to other electrochemical biosensors. This study is the first to report on an extended-gate type organic field effect transistor (OFET) for lactate detection in aqueous media. Here, the extended-gate electrode of the OFET was modified with layers of a lactate oxidase and a horseradish peroxidase osmium-redox polymer on a flexible plastic film substrate for an enzymatic redox reaction of lactate. The device exhibited both high selectivity and sensitivity. The limit of detection (LOD) and the limit of quantification (LOQ) were estimated to be 66 nM and 220 nM, respectively, which are the sufficient detection limit for practical sensor applications. The obtained results confirm that extended-gate type OFET devices are applicable to enzyme-based biosensors for detecting lactate levels.
Assuntos
Técnicas Biossensoriais/instrumentação , Lactatos/análise , Osmio/química , Polímeros/química , Transistores Eletrônicos , Eletrodos , Enzimas Imobilizadas/química , Desenho de Equipamento , Limite de Detecção , Oxigenases de Função Mista/química , Água/análiseRESUMO
We examined the correlation between the glomerular filtration rate (GFR) estimated from an equation based on the serum iodixanol clearance technique and International Renal Interest Society (IRIS) stages of chronic kidney disease (CKD) in cats. The equation included the injection dose, sampling time, serum concentration and estimated volume of distribution (Vd) of the isotonic, nonionic, contrast medium iodixanol as a test tracer. The percent changes in the median basal GFR values calculated from the equation in CKD cats resembled those of IRIS stages 1-3. These data validate the association between the GFR derived from the simplified equation and IRIS stages based on the serum creatinine concentration in cats with CKD. They describe the GFR ranges determined using single-sample iodixanol clearance for healthy cats and cats with various IRIS stages of CKD.
Assuntos
Doenças do Gato/diagnóstico , Meios de Contraste/farmacocinética , Taxa de Filtração Glomerular , Insuficiência Renal Crônica/veterinária , Ácidos Tri-Iodobenzoicos/farmacocinética , Animais , Estudos de Casos e Controles , Doenças do Gato/sangue , Doenças do Gato/fisiopatologia , Gatos , Creatinina/sangue , Feminino , Taxa de Filtração Glomerular/fisiologia , Masculino , Insuficiência Renal Crônica/sangue , Insuficiência Renal Crônica/diagnóstico , Insuficiência Renal Crônica/fisiopatologia , Índice de Gravidade de Doença , Ácidos Tri-Iodobenzoicos/sangueRESUMO
To testify the relevance of Jacobsson's equation for estimating bovine glomerular filtration rate (GFR), we prepared an integrated formula based on its equation using clinically healthy dairy (n=99) and beef (n=63) cows, and cows with reduced renal function (n=15). The isotonic, nonionic, contrast medium iodixanol was utilized as a test tracer. The GFR values estimated from the integrated formula were well consistent with those from the standard multisample method in each cow strain, and the Holstein equation prepared by a single blood sample in Holstein dairy cows. The basal reference GFR value in healthy dairy cows was significantly higher than that in healthy beef cows, presumably due to a breed difference or physiological state difference. It is concluded that the validity for the application of Jacobsson's equation to estimate bovine GFR is proven and it can be used in bovine practices.
Assuntos
Bovinos/fisiologia , Meios de Contraste , Taxa de Filtração Glomerular , Ácidos Tri-Iodobenzoicos , Animais , Feminino , Soluções IsotônicasRESUMO
To develop an expedient procedure for estimating glomerular filtration rate (GFR) in conscious mice, we first established a simple technique for repeated blood collections from the tail vein using a microcapillary tube attached to a 24-gauge needle without the hub. Then, we devised a definition equation for estimation of the GFR using the contrast medium iodixanol as a test tracer. Iodixanol was administered as a bolus injection at 1500 mg I/kg to ddY mice, and the GFR was determined by the conventional multisample strategy. Based on cumulative data from the multisample method, an equation for the single-blood-sample method including the iodixanol dose, estimated distribution volume (Vd), and plasma iodixanol concentration at 60 min later was sought. The GFR values from the multisample method were in good agreement with those calculated using the equation. In clinically healthy mice, the GFR decreased gradually with ages from 11 weeks old in both sexes, suggesting the necessity of the corresponding control in each protocol. In nephropathy mice induced by cisplatin, the GFR values decreased with rises in serum BUN and creatinine concentrations, and serum creatinine became elevated when the GFR decreased to approximately 70% of the basal value. The results suggest that the simplified equation using iodixanol, allowing for the repeated use of the same mice, is a versatile procedure for research purposes.
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To estimate the glomerular filtration rate (GFR) in cynomolgus monkeys (Macaca fascicularis), a three-blood-sample method using iodixanol was assessed in comparison with the conventional multisample strategy using inulin. Iodixanol and inulin were coadministered intravenously 40 mg I/kg and 50 mg/kg, respectively, to male monkeys, followed by blood collection 60, 90 and 120 min later. A close correlation (r=0.96) was noted between the GFR values estimated by both methods. In clinically healthy monkeys, the basal values were determined to be 3.06 ± 0.50 ml/min/kg. This is the first report, suggesting that serum clearance of iodixanol is a ready-to-use tool for a screening the GFR in monkeys, although it is necessary to perform a more longitudinal study using animals with reduced renal function.
Assuntos
Taxa de Filtração Glomerular/fisiologia , Macaca fascicularis/fisiologia , Animais , Meios de Contraste/farmacologia , Relação Dose-Resposta a Droga , Inulina/farmacologia , Masculino , Valores de Referência , Ácidos Tri-Iodobenzoicos/farmacologiaRESUMO
Influenza virus is rich in variation and mutations. It would be very convenient for virus detection and isolation to histochemically detect viral infection regardless of variation and mutations. Here, we established a histochemical imaging assay for influenza virus sialidase activity in living cells by using a new fluorescent sialidase substrate, 2-(benzothiazol-2-yl)-4-bromophenyl 5-acetamido-3,5-dideoxy-α-D-glycero-D-galacto-2-nonulopyranosidonic acid (BTP3-Neu5Ac). The BTP3-Neu5Ac assay histochemically visualized influenza virus-infected cells regardless of viral hosts and subtypes. Influenza virus neuraminidase-expressed cells, viral focus formation, and virus-infected locations in mice lung tissues were easily, rapidly, and sensitively detected by the BTP3-Neu5Ac assay. Histochemical visualization with the BTP3-Neu5Ac assay is extremely useful for detection of influenza viruses without the need for fixation or a specific antibody. This novel assay should greatly improve the efficiency of detection, titration, and isolation of influenza viruses and might contribute to research on viral sialidase.