Impacts of Fuel Stage Ratio on the Morphological and Nanostructural Characteristics of Soot Emissions from a Twin Annular Premixing Swirler Combustor.

Soot particles emitted from aircraft engines constitute a major anthropogenic source of pollution in the vicinity of airports and at cruising altitudes. This emission poses a significant threat to human health and may alter the global climate. Understanding the characteristics of soot particles, particularly those generated from Twin Annular Premixing Swirler (TAPS) combustors, a mainstream combustor in civil aviation engines, is crucial for aviation environmental protection. In this study, a comprehensive characterization of soot particles emitted from TAPS combustors was conducted using scanning electron microscopy (SEM), high-resolution transmission electron microscopy (HRTEM), and Raman spectroscopy. The morphology and nanostructure of soot particles were examined across three distinct fuel stage ratios (FSR), at 10%, 15%, and 20%. The SEM analysis of soot particle morphology revealed that coated particles constitute over 90% of the total particle sample, with coating content increasing proportionally to the fuel stage ratio. The results obtained from HRTEM indicated that average primary particle sizes increase with the fuel stage ratio. The results of HRTEM and Raman spectroscopy suggest that the nanostructure of soot particles becomes more ordered and graphitized with an increasing fuel stage ratio, resulting in lower oxidation activity. Specifically, soot fringe length increased with the fuel stage ratio, while soot fringe tortuosity and separation distance decreased. In addition, there is a prevalent occurrence of defects in the graphitic lattice structure of soot particles, suggesting a high degree of elemental carbon disorder.

Genome-Wide Identification of the SPP/SPPL Gene Family and BnaSPPL4 Regulating Male Fertility in Rapeseed (Brassica napus L.).

Signal peptide peptidase (SPP) and its homologs, signal peptide peptidase-like (SPPL) proteases, are members of the GxGD-type aspartyl protease family, which is widespread in plants and animals and is a class of transmembrane proteins with significant biological functions. SPP/SPPLs have been identified; however, the functions of SPP/SPPL in rapeseed (Brassica napus L.) have not been reported. In this study, 26 SPP/SPPLs were identified in rapeseed and categorized into three groups: SPP, SPPL2, and SPPL3. These members mainly contained the Peptidase_A22 and PA domains, which were distributed on 17 out of 19 chromosomes. Evolutionary analyses indicated that BnaSPP/SPPLs evolved with a large number of whole-genome duplication (WGD) events and strong purifying selection. Members are widely expressed and play a key role in the growth and development of rapeseed. The regulation of rapeseed pollen fertility by the BnaSPPL4 gene was further validated through experiments based on bioinformatics analysis, concluding that BnaSPPL4 silencing causes male sterility. Cytological observation showed that male infertility caused by loss of BnaSPPL4 gene function occurs late in the mononucleate stage due to microspore dysplasia.

Mechanistic Study on the Species Differences in Excretion Pathway of HR011303 in Humans and Rats.

Excretion of [14C]HR011303-derived radioactivity showed significant species difference. Urine (81.50% of dose) was the main excretion route in healthy male subjects, whereas feces (87.16% of dose) was the main excretion route in rats. To further elucidate the underlying cause for excretion species differences of HR011303, studies were conducted to uncover its metabolism and excretion mechanism. M5, a glucuronide metabolite of HR011303, is the main metabolite in humans and rats. Results of a rat microsome incubation study suggested that HR011303 was metabolized to M5 in the rat liver. According to previous studies, M5 is produced in both human liver and kidney microsomes. We found that M5 in the human liver can be transported to the blood by multidrug resistance-associated protein (MRP) 3, and then the majority of M5 can be hydrolyzed to HR011303. HR011303 enters the human kidney or liver through passive diffusion, whereas M5 is taken up through organic anion transporter (OAT) 3, organic anion-transporting polypeptide (OATP) 1B1, and OATP1B3. When HR011303 alone is present, it can be metabolized to M5 in both sandwich-cultured rat hepatocytes (SCRH) and sandwich-cultured human hepatocytes (SCHH) and excreted into bile as M5 in SCRH. Using transporter inhibitors in sandwich-cultured model and membrane vesicles expressing MRP2 or Mrp2, we found that M5 was a substance of MRP2/Mrp2, and the bile efflux of M5 was mainly mediated by MRP2/Mrp2. Considering the significant role of MRP3/Mrp3 and MRP2/Mrp2 in the excretion of glucuronides, the competition between them for M5 was possibly the determinant for the different excretion routes in humans and rats. SIGNIFICANCE STATEMENT: Animal experiments are necessary to predict dosage and safety of candidate drugs prior to clinical trials. However, extrapolation results often differ from the actual situation. For HR011303, excretory pathways exhibited a complete reversal, through urine in humans and feces in rats. Such phenomena have been observed in several drugs, but no in-depth studies have been conducted to date. In the present study, the excretion species differences of HR011303 can be explained by the competition for M5 between MRP2/Mrp2 and MRP3/Mrp3.

Pharmacokinetics, Mass Balance, and Metabolism of the Novel Urate Transporter 1 Inhibitor [14C]HR011303 in Humans: Metabolism Is Mediated Predominantly by UDP-Glucuronosyltransferase.

HR011303, a promising selective urate transporter 1 inhibitor, is currently being studied in a phase III clinical trial in China for the treatment of hyperuricemia and gout. In the current study, the pharmacokinetics, mass balance, and metabolism of HR011303 were examined in six healthy Chinese male subjects who received a single oral dose of 10 mg of [14C]HR011303 (80 µCi). The results showed that HR011303 was rapidly absorbed with a median time to reach C max of 1.50 hours postdose, and the arithmetic mean half-life of total radioactivity was approximately 24.2 hours in plasma. The mean blood-to-plasma radioactivity concentration ratio was 0.66, suggesting the preferential distribution of drug-related components in plasma. At 216 hours postdose, the mean cumulative excreted radioactivity was 91.75% of the dose, including 81.50% in urine and 10.26% in feces. Six metabolites were identified, and the parent drug HR011303 was the most abundant component in plasma and feces, but a minor component in urine. Glucuronidation of the carboxylic acid moiety of HR011303 was the primary metabolic pathway in humans, amounting to 69.63% of the dose (M5, 51.57% of the dose; M5/2, 18.06% of the dose) in the urine; however, it was not detected in plasma. UDP-glucuronosyltransferase (UGT) 2B7 was responsible for the formation of M5. Overall, after a single oral dose of 10 mg of [14C]HR011303 (80 µCi), HR011303 and its main metabolites were eliminated via renal excretion. The major metabolic pathway was carboxylic acid glucuronidation, which was catalyzed predominantly by UGT2B7. SIGNIFICANCE STATEMENT: This study determined the absorption and disposition of HR011303, a selective urate transporter (URAT) 1 inhibitor currently in development for the treatment of hyperuricemia and gout. This work helps to characterize the major metabolic pathways of new URAT inhibitors and identify the absorption and clearance mechanism.

Metabolite profiles and mass balance of fuzuloparib, a novel poly (ADP-ribose) polymerase inhibitor, in subjects with advanced solid cancers.

AIM: This trial (NCT04013048) investigated the metabolite profiles, mass balance and pharmacokinetics of fuzuloparib, a novel poly (ADP-ribose) polymerase inhibitor, in subjects with advanced solid cancers. METHODS: A single dose of 150 mg [14 C]fuzuloparib was administered to five subjects with advanced solid cancers. Blood, urine and faecal samples were collected, analysed for radioactivity and unchanged fuzuloparib, and profiled for metabolites. The safety of the medicine was assessed during the study. RESULTS: The maximum concentrations (Cmax ) of the total radioactivity (TRA) and unchanged fuzuloparib in plasma were 5.39 µg eq./mL and 4.19 µg/mL, respectively, at approximately 4 hours post dose. The exposure (AUC0-t ) of fuzuloparib accounted for 70.7% of the TRA in plasma, and no single metabolite was observed accounting for more than 10% of the plasma TRA. The recovery of TRA in excreta was 103.3 ± 3.8% in 288 hours, including 59.1 ± 9.9% in urine and 44.2 ± 10.8% in faeces. Sixteen metabolites of fuzuloparib were identified, including mono-oxidation (M1), hydrogenation (M2), di-oxidation (M3), trioxidation (M4), glucuronidation (M5, M7, M8) and de-ethylation (M6) products, and there was no specific binding between these metabolites and blood cells. Aliphatic hydroxylated fuzuloparib (M1-1) was the primary metabolite in the excreta, accounting for more than 40% of the dose for subjects. There were no serious adverse events observed in the study. CONCLUSION: Fuzuloparib was widely metabolized and excreted completely through urine and faeces in subjects with advanced solid cancer. Unchanged fuzuloparib was indicated to be the primary drug-related compound in circulation. [14 C]fuzuloparib was well-tolerated at the study dose.

Complete genome sequence of passiflora virus Y infecting passion fruit in China.

The complete genome sequence of passiflora virus Y (PaVY) from passion fruit growing in Guangdong province, China, was determined. The entire positive single-strand RNA genome comprises 9681 nucleotides (nt) excluding the poly(A) tail and encodes a polyprotein of 3084 amino acids flanked by 5' and 3' untranslated regions of 169 and 257 nt, respectively. In sequence comparisons and phylogenetic analysis, PaVY appears to represent a new species in the bean common mosaic virus subgroup of the genus Potyvirus. This is the first report of the complete genome sequence of PaVY and the first report of this virus in China.

Phase I dose-escalation and expansion study of PARP inhibitor, fluzoparib (SHR3162), in patients with advanced solid tumors.

OBJECTIVE: Fluzoparib (SHR3162) is a novel, potent poly(ADP-ribose) polymerases (PARP)1, 2 inhibitor that showed anti-tumor activity in xenograft models. We conducted a phase I, first-in-human, dose-escalation and expansion (D-Esc and D-Ex) trial in patients with advanced solid cancer. METHODS: This was a 3+3 phase I D-Esc trial with a 3-level D-Ex at 5 hospitals in China. Eligible patients for D-Esc had advanced solid tumors refractory to standard therapies, and D-Ex enrolled patients with ovarian cancer (OC). Fluzoparib was administered orally once or twice daily (bid) at 11 dose levels from 10 to 400 mg/d. Endpoints included dose-finding, safety, pharmacokinetics, and antitumor activity. RESULTS: Seventy-nine patients were enrolled from March, 2015 to January, 2018 [OC (47, 59.5%); breast cancer (BC) (16, 20.3%); colorectal cancer (8, 10.1%), other tumors (8, 10.1%)]; 48 patients were treated in the D-Esc arm and 31 in the D-Ex arm. The maximum tolerated dose (MTD) was 150 mg bid, with a half-life of 9.14 h. Grade 3/4 adverse events included anemia (7.6%) and neutropenia (5.1%). The objective response rate (ORR) was 30% (3/10) in patients with platinum-sensitive OC and 7.7% (1/13) in patients with BC. Among patients treated with fluzoparib ≥120 mg/d, median progression-free survival (mPFS) was 7.2 [95% confidence interval (95% CI), 1.8-9.3] months in OC, 9.3 (95% CI, 7.2-9.3) months in platinum-sensitive OC, and 3.5 (range, 2.0-28.0) months in BC. In patients with germline BC susceptibility gene mutation (gBRCA Mut) (11/43 OC; 2/16 BC), mPFS was 8.9 months for OC (range, 1.0-23.2; 95% CI, 1.0-16.8) and 14 and 28 months for BC (those two patients both also had somaticBRCA Mut). CONCLUSIONS: The MTD of fluzoparib was 150 mg bid in advanced solid malignancies. Fluzoparib demonstrated single-agent antitumor activity in BC and OC, particularly in BRCA Mut and platinum-sensitive OC.

Highly efficient fluoride adsorption from aqueous solution by nepheline prepared from kaolinite through alkali-hydrothermal process.

A direct alkali-hydrothermal induced transformation process was adopted to prepare nepheline from raw kaolinite (shortened form RK in this paper) and NaOH solution in this paper. Structure and morphology characterizations of the synthetic product showed that the nepheline possessed high degree of crystallinity and uniform surface morphology. Specific surface area of nepheline is 18 m2/g, with a point of zero charge at around pH 5.0-5.5. The fluoride (F- ions) adsorption by the synthetic nepheline (shortened form SN in this paper) from aqueous solution was also investigated under different experimental conditions. The adsorption process well matched the Langmuir isotherm model with an amazing maximum adsorption capacity of 183 mg/g at 323 K. The thermodynamic parameters (ΔG0, ΔH0, and ΔS0) for adsorption on SN were also determined from the temperature dependence. The adsorption capacities of fluoride on SN increased with increasing of temperature and initial concentration. Initial pH value also had influence on adsorption process. Adsorption of fluoride was rapidly increased in 5-60 min and thereafter increased slowly to reach the equilibrium in about 90-180 min under all conditions. The adsorption followed a pseudo-second order rate law.

Mediating Effects of Working Memory in the Relation Between Rapid Automatized Naming and Chinese Reading Comprehension.

This study examined the mediating role of working memory (WM) in the relation between rapid automatized naming (RAN) and Chinese reading comprehension. Three tasks assessing differentially visual and verbal components of WM were programmed by E-prime 2.0. Data collected from 55 Chinese college students were analyzed using correlations and hierarchical regression methods to determine the connection among RAN, reading comprehension, and WM components. Results showed that WM played a significant mediating role in the RAN-reading relation and that auditory WM made stronger contributions than visual WM. Taking into account of the multi-component nature of WM and the specificity of Chinese reading processing, this study discussed the mediating powers of the WM components, particularly auditory WM, further clarifying the possible components involved in the RAN-reading relation and thus providing some insight into the complicated Chinese reading process.

Assessing the particulate matter emission reduction characteristics of small turbofan engine fueled with 100 % HEFA sustainable aviation fuel.

With the continuous increase in global air transportation, the impact of ultrafine particulate matter (PM) emissions from aviation on human health and environmental pollution is becoming increasingly severe. In addition to carbon reduction throughout the lifecycle, Sustainable Aviation Fuels (SAF) also represent a significant pathway for reducing PM emissions. However, due to issues such as airworthiness safety and adaptability, existing research has mostly focused on the emission performance of SAF when blended with traditional fuels at <50 %, leaving the emission characteristics of higher blending ratios to be explored. In this study, using measurement methods recommended by the International Civil Aviation Organization (ICAO), the PM emission reduction characteristics of small turbofan engines fueled with 100 % Hydroprocessed Esters and Fatty Acids (HEFA)-SAF were experimentally evaluated and compared with traditional fuels RP-3 and Diesel, while avoiding the interference of lubricant blending combustion. The results showed that the peak number concentration of particle size distribution (PSD), PM total number, as well as the number and mass concentration of non-volatile particulate matter (nvPM) decreased initially and then increased with rising thrust conditions. HEFA-SAF exhibits PSD with smaller diameters, and the Geometric Mean Diameter (GMD) ranges from 7.7 nm to 20.3 nm under all conditions. Both volatile particulates (vPM) and nvPM from HEFA-SAF are significantly reduced, with nvPM number emission index (EIn) being 92 % and 71 % lower than Diesel and RP-3, respectively. The nvPM mass emission index (EIm) also shows reductions of 96 % and 89 % compared to Diesel and RP-3. Microscopic characterization also indicated that using HEFA-SAF emitted fewer and smaller PMs. This study establishes a foundation for evaluating the effectiveness of 100 % SAF in reducing PM emissions within the aviation sector, and contributes to the airworthiness regulations development related to the use of SAF in a variety of application environments, alongside enhancing environmental protection measures.

A convolutional neural network prediction model for aviation nitrogen oxides emissions throughout all flight phases.

In recent years, there has been an increasing amount of research on nitrogen oxides (NOx) emissions, and the environmental impact of aviation NOx emissions at cruising altitudes has received widespread attention. NOx may play a crucial role in altering the composition of the atmosphere, particularly regarding ozone formation in the upper troposphere. At present, the ground emission database based on the landing and takeoff (LTO) cycle is more comprehensive, while high-altitude emission data is scarce due to the prohibitively high cost and the inevitable measurement uncertainty associated with in-flight sampling. Therefore, it is necessary to establish a comprehensive NOx emission database for the entire flight envelope, encompassing both ground and cruise phases. This will enable a thorough assessment of the impact of aviation NOx emissions on climate and air quality. In this study, a prediction model has been developed via convolutional neural network (CNN) technology. This model can predict the ground and cruise NOx emission index for turbofan engines and mixed turbofan engines fueled by either conventional aviation kerosene or sustainable aviation fuels (SAFs). The model utilizes data from the engine emission database (EEDB) released by the International Civil Aviation Organization (ICAO) and results obtained from several in-situ emission measurements conducted during ground and cruise phases. The model has been validated by comparing measured and predicted data, and the results demonstrate its high prediction accuracy for both the ground (R2 > 0.95) and cruise phases (R2 > 0.9). This surpasses traditional prediction models that rely on fuel flow rate, such as the Boeing Fuel Flow Method 2 (BFFM2). Furthermore, the model can predict NOx emissions from aircrafts burning SAFs with satisfactory accuracy, facilitating the development of a more complete and accurate aviation NOx emission inventory, which can serve as a basis for aviation environmental and climatic research. SYNOPSIS: The utilization of the ANOEPM-CNN offers a foundation for establishing more precise emission inventories, thereby reducing inaccuracies in assessing the impact of aviation NOx emissions on climate and air quality.

Tomato based gelatin methacryloyl hydrogel as an effective natural and low-cost scaffold for accelerative wound healing.

Oxidative stress and infection are the main reasons for postponement of wound healing rate. They can potentially lead to serious inflammation and eventually lead to a longer and more painful recovery phase. Although wound dressings based on synthetic materials with antioxidative property have been proved to exhibit remarkable effect in controlling ROS level and improving wound healing, issues, such as high cost in raw materials, complicated procedures, usage of various toxic additives, and potential allergies, have significantly confined further clinical applications. In this study, a novel type of tissue engineering scaffold, based on tomatoes (Solanum lycopersicon) and gelatin methacryloyl (GelMA), was prepared via facile lyophilization and photo cross-link method (SL/GelMA). By taking advantages of various antioxidative components, such as carotenoids, flavonoids, phenolic acids, vitamin E, and vitamin C in tomatoes, SL/GelMA can effectively regulate ROS level, relieve the oxidative stress in wound bed, promote cell migration and angiogenesis, contribute to collagen deposition, and thus accelerate the rate of wound enclosure. Along with its high biocompatibility and low allergic potential, we believe that the food-derived wound dressing with facile preparation method, easy accessibility, and high cost-effectiveness can be translated for clinical treatments of various chronic wounds.

Preparation of Salicylic Acid-Functionalized Nanopesticides and Their Applications in Enhancing Salt Stress Resistance.

Soil salinization is one of the global ecological and environmental problems that are tremendously threatening to the sustainable development of agriculture and food supply. In this work, a facile strategy was proposed to enhance the salt stress resistance of plants by preparing salicylic acid (SA)-functionalized mesoporous silica nanocarriers loaded with emamectin benzoate (EB). The obtained nanopesticides demonstrated a particle size of less than 300 nm. As an endogenous plant hormone, the grafting of SA in this nanopesticide system improved the uptake and translocation of pesticides in cucumber plants by 145.06%, and the applications of such nanopesticides enhanced the salt stress resistance of plants. This phenomenon was accounted for by the SA-functionalized nanopesticides increasing the superoxide dismutase and peroxidase activities (640 and 175%, respectively) and reducing the malondialdehyde content (54.10%), correspondingly alleviating the accumulation of reactive oxygen species and cell damage in plants. The above results were also confirmed by Evans blue staining and NBT staining experiments on cucumber leaves. In addition, these nanopesticides exhibited high insecticidal toxicity, and they also demonstrated biosafety toward nontarget organisms due to their sustained release property. Therefore, this work developed a biosafe SA-functionalized nanopesticide system, and these newly developed nanopesticides have potential in the agricultural field for enhancing salt stress resistance of plants.

A mass balance study of [14C]SHR6390 (dalpiciclib), a selective and potent CDK4/6 inhibitor in humans.

SHR6390 (dalpiciclib) is a selective and effective cyclin-dependent kinase (CDK) 4/6 inhibitor and an effective cancer therapeutic agent. On 31 December 2021, the new drug application was approved by National Medical Product Administration (NMPA). The metabolism, mass balance, and pharmacokinetics of SHR6390 in 6 healthy Chinese male subjects after a single oral dose of 150 mg [14C]SHR6390 (150 µCi) in this research. The Tmax of SHR6390 was 3.00 h. In plasma, the t 1/2 of SHR6390 and its relative components was approximately 17.50 h. The radioactivity B/P (blood-to-plasma) AUC0-t ratio was 1.81, indicating the preferential distribution of drug-related substances in blood cells. At 312 h after administration, the average cumulative excretion of radioactivity was 94.63% of the dose, including 22.69% in urine and 71.93% in stool. Thirteen metabolites were identified. In plasma, because of the low level of radioactivity, only SHR6390 was detected in pooled AUC0-24 h plasma. Stool SHR6390 was the main component in urine and stool. Five metabolites were identified in urine, and 12 metabolites were identified in stool. Overall, faecal clearance is the main method of excretion.

Ischemia-induced brain damage is enhanced in human renin and angiotensinogen double-transgenic mice.

To investigate the role of brain angiotensin II (ANG II) in the pathogenesis of injury following ischemic stroke, mice overexpressing renin and angiotensinogen (R+A+) and their wild-type control animals (R-A-) were used for experimental ischemia studies. Focal brain ischemia was induced by middle cerebral artery occlusion (MCAO). The severity of ischemic injury was determined by measuring neurological deficits and histological damage at 24 and 48 h after MCAO, respectively. To exclude the influence of blood pressure and local collateral blood flow, brain slices were used for oxygen and glucose deprivation (OGD) studies. The severity of OGD-induced damage was determined by measuring indicators of tissue swelling and cell death, the intensity of the intrinsic optical signal (IOS), and the number of propidium iodide (PI) staining cells, respectively. Results showed 1) R+A+ mice showed higher neurological deficit score (3.8 +/- 0.5 and 2.5 +/- 0.3 for R+A+ and R-A-, respectively, P < 0.01) and larger infarct volume (22.2 +/- 1.6% and 14.1 +/- 1.2% for R+A+ and R-A-, respectively, P < 0.01); 2) The R+A+ brain slices showed more severe tissue swelling and cell death in the cortex (IOS: 140 +/- 6% and 114 +/- 10%; PI: 139 +/- 20 cells/field and 39 +/- 9 cells/field for R+A+ and R-A-, respectively, P < 0.01); 3) treatment with losartan (20 micromol/l) abolished OGD-induced exaggeration of cell injury seen in R+A+ mice. The data indicate that activation of ANG II/AT(1) signaling is harmful to brain exposed to ischemia.

Antioxidant Enzyme-Mimetic Activity and Neuroprotective Effects of Cerium Oxide Nanoparticles Stabilized with Various Ratios of Citric Acid and EDTA.

Cerium oxide (CeO2) nanoparticles (CeNPs) are potent antioxidants that are being explored as potential therapies for diseases in which oxidative stress plays an important pathological role. However, both beneficial and toxic effects of CeNPs have been reported, and the method of synthesis as well as physico-chemical, biological, and environmental factors can impact the ultimate biological effects of CeNPs. In the present study, we explored the effect of different ratios of citric acid (CA) and EDTA (CA/EDTA), which are used as stabilizers during synthesis of CeNPs, on the antioxidant enzyme-mimetic and biological activity of the CeNPs. We separated the CeNPs into supernatant and pellet fractions and used commercially available enzymatic assays to measure the catalase-, superoxide dismutase (SOD)-, and oxidase-mimetic activity of each fraction. We tested the effects of these CeNPs in a mouse hippocampal brain slice model of ischemia to induce oxidative stress where the fluorescence indicator SYTOX green was used to assess cell death. Our results demonstrate that CeNPs stabilized with various ratios of CA/EDTA display different enzyme-mimetic activities. CeNPs with intermediate CA/EDTA stabilization ratios demonstrated greater neuroprotection in ischemic mouse brain slices, and the neuroprotective activity resides in the pellet fraction of the CeNPs. The neuroprotective effects of CeNPs stabilized with equal proportions of CA/EDTA (50/50) were also demonstrated in two other models of ischemia/reperfusion in mice and rats. Thus, CeNPs merit further development as a neuroprotective therapy for use in diseases associated with oxidative stress in the nervous system.

Calcium/calmodulin-modulated chloride and taurine conductances in cultured rat astrocytes.

Osmotically swollen rat cerebral astrocytes develop an increased anion conductance which can mediate chloride and taurine release. We used whole cell patch clamp to study mechanisms that modulate this conductance. Astrocyte chloride conductance increased within 4 min of exposure to 200 mOsm medium and was 670+/-123% of its initial value after 15 min (mean+/-S.E.M.). This conductance was substantially reduced in 0.1 mM extracellular calcium with 20 mM BAPTA added to the electrode solution and was completely inhibited with calcium-free perfusion solution containing 1 mM EDTA (n=4). The conductance increase in 200 mOsm medium also was inhibited in a dose-dependent manner by nimodipine with a calculated K(i) of 0.31+/-0.4 microM and mean+/-S.E.M. inhibition of 84.4+/-4% at 100 microM nimodipine. In the presence of 100 microM W-7, a calmodulin antagonist, the mean+/-S.E.M. conductance increase after 15 min was 223+/-40% of the initial value while 300 microM W-7 or 100 microM trifluoperazine inhibited the conductance increase completely (n=6). With taurine as the major anion in electrode and perfusion solutions, a significant conductance increase was observed in 200 mOsm medium. This conductance increase was inhibited by 300 microM W-7 or 100 microM nimodipine. We conclude extracellular calcium influx via L-type calcium channels leads to increased astrocyte anion conductance in 200 mOsm conditions via calmodulin-dependent activation of anion channels. Efflux of anionic taurine from swollen astrocytes also may be affected by calcium influx through a similar calcium/calmodulin-dependent process.

Angiotensin converting enzyme 2/Ang-(1-7)/mas axis protects brain from ischemic injury with a tendency of age-dependence.

BACKGROUND: The angiotensin (Ang) converting enzyme 2 (ACE2)/Ang-(1-7)/Mas receptor pathway is an important component of the renin-angiotensin system and has been suggested to exert beneficial effects in ischemic stroke. AIMS: This study explored whether the ACE2/Ang-(1-7)/Mas pathway has a protective effect on cerebral ischemic injury and whether this effect is affected by age. METHODS: We used three-month and eight-month transgenic mice with neural over-expression of ACE2 (SA) and their age-matched nontransgenic (NT) controls. Neurological deficits and ischemic stroke volume were determined following middle cerebral artery occlusion (MCAO). In oxygen and glucose deprivation (OGD) experiments on brain slices, the effects of the Mas receptor agonist (Ang1-7) or antagonist (A779) on tissue swelling, Nox2/Nox4 expression reactive oxygen species (ROS) production and cell death were measured. RESULTS: (1) Middle cerebral artery occlusion -induced ischemic injury and neurological deficit were reduced in SA mice, especially in eight-month animals; (2) OGD-induced tissue swelling and cell death were decreased in SA mice with a greater reduction seen in eight-month mice; (3) Ang-(1-7) and A779 had opposite effects on OGD-induced responses, which correlated with changes in Nox2/Nox4 expression and ROS production. CONCLUSIONS: Angiotensin converting enzyme 2/Ang-(1-7)/Mas axis protects brain from ischemic injury via the Nox/ROS signaling pathway, with a greater effect in older animals.

BMP7 induces the differentiation of bone marrow-derived mesenchymal cells into chondrocytes.

Injured articular cartilage has a poor capacity for spontaneous healing. So far, a satisfactory solution to repair the injured cartilage has not been found, but transgenic therapy might be a promising treatment. This study aims to evaluate the potential of transfecting bone morphogenetic protein-7 (BMP-7), a secretory protein, into bone marrow-derived mesenchymal stem cells (BMSCs), in inducing the differentiation of bone marrow stromal cells into chondrocytes in vitro. The phenotypes of the cells were observed by alcian blue staining and H&E staining with an inverted microscope. The glycosaminoglycan (GAG) content of BMSCs transfected with pcDNA3.1-BMP7 or induced by inducing medium was examined after 7, 14, or 21 days of incubation. A standard curve as reference for BMSCs' GAG content was plotted using galacturonic acid. The content of type II collagen in culture medium was detected by ELISA. Our results demonstrated that BMP7-transfected BMSCs or BMSCs incubated with inducing medium possess the ability to differentiate into chondrocytes. BMP7-induced BMSCs secrete type II collagen and GAG. There was no significant difference between BMP7-induced BMSCs in their secreted protein content when compared with the positive control group (TGF-ß1 and dexamethasone) (P > 0.05), but there was significant difference in the secreted protein profile when compared with the negative control group (P < 0.05).

Purinergic activation of anion conductance and osmolyte efflux in cultured rat hippocampal neurons.

The majority of mammalian cells demonstrate regulatory volume decrease (RVD) following swelling caused by hyposmotic exposure. A critical signal initiating RVD is activation of nucleotide receptors by ATP. Elevated extracellular ATP in response to cytotoxic cell swelling during pathological conditions also may initiate loss of taurine and other intracellular osmolytes via anion channels. This study characterizes neuronal ATP-activated anion current and explores its role in net loss of amino acid osmolytes. To isolate anion currents, we used CsCl as the major electrolyte in patch electrode and bath solutions and blocked residual cation currents with NiCl(2) and tetraethylammonium. Anion currents were activated by extracellular ATP with a K(m) of 70 microM and increased over fourfold during several minutes of ATP exposure, reaching a maximum after 9.0 min (SD 4.2). The currents were blocked by inhibitors of nucleotide receptors and volume-regulated anion channels (VRAC). Currents showed outward rectification and inactivation at highly depolarizing membrane potentials, characteristics of swelling-activated anion currents. P2X agonists failed to activate the anion current, and an inhibitor of P2X receptors did not block the effect of ATP. Furthermore, current activation was observed with extracellular ADP and 2-(methylthio)adenosine 5'-diphosphate, a P2Y(1) receptor-specific agonist. Much less current activation was observed with extracellular UTP, suggesting the response is mediated predominantly by P2Y(1) receptors. ATP caused a dose-dependent loss of taurine and alanine that could be blocked by inhibitors of VRAC. ATP did not inhibit the taurine uptake transporter. Thus extracellular ATP triggers a loss of intracellular organic osmolytes via activation of anion channels. This mechanism may facilitate neuronal volume homeostasis during cytotoxic edema.