Electrophysiological Changes in the Rabbit Ventricular Wedge and Human-Induced Pluripotent Stem-Cell Derived (IPSC) Cardiomyocytes Translate to Severe Arrhythmia Observed in a Canine Toxicology Study, Not Predicted by Standard In Vitro Ion Channel Assays.

During drug discovery, small molecules are typically assayed in vitro for secondary pharmacology effects, which include ion channels relevant to cardiac electrophysiology. Compound A was an irreversible inhibitor of myeloperoxidase investigated for the treatment of peripheral artery disease. Oral doses in dogs at ≥5 mg/kg resulted in cardiac arrhythmias in a dose-dependent manner (at Cmax, free ≥1.53 µM) that progressed in severity with time. Nevertheless, a panel of 13 different cardiac ion channel (K, Na, and Ca) assays, including hERG, failed to identify pharmacologic risks of the molecule. Compound A and a related Compound B were evaluated for electrophysiological effects in the isolated rabbit ventricular wedge assay. Compounds A and B prolonged QT and Tp-e intervals at ≥1 and ≥.3 µM, respectively, and both prolonged QRS at ≥5 µM. Compound A produced early after depolarizations and premature ventricular complexes at ≥5 µM. These data indicate both compounds may be modulating hERG (Ikr) and Nav1.5 ion channels. In human IPSC cardiomyocytes, Compounds A and B prolonged field potential duration at ≥3 µM and induced cellular dysrhythmia at ≥10 and ≥3 µM, respectively. In a rat toxicology study, heart tissue: plasma concentration ratios for Compound A were ≥19X at 24 hours post-dose, indicating significant tissue distribution. In conclusion, in vitro ion channel assays may not always identify cardiovascular electrophysiological risks observed in vivo, which can be affected by tissue drug distribution. Risk for arrhythmia may increase with a "trappable" ion channel inhibitor, particularly if cardiac tissue drug levels achieve a critical threshold for pharmacologic effects.

The incidence of spontaneous arrhythmias in telemetered beagle dogs, Göttingen Minipigs and Cynomolgus non-human primates: A HESI consortium retrospective analysis.

INTRODUCTION: Characterization of the incidence of spontaneous arrhythmias to identify possible drug-related effects is often an important part of the analysis in safety pharmacology studies using telemetry. METHODS: A retrospective analysis in non-clinical species with and without telemetry transmitters was conducted. Electrocardiograms (24 h) from male and female beagle dogs (n = 131), Göttingen minipigs (n = 108) and cynomolgus non-human primates (NHP; n = 78) were analyzed. RESULTS: Ventricular tachycardia (VT) was observed in 3% of the dogs but was absent in minipigs and NHPs. Ventricular fibrillation (VF) was not observed in the 3 species. Ventricular premature beats (VPBs) were more frequent during daytime and atrioventricular blocks (AVBs) were more frequent at night in all species. A limited number of animals exhibited a high arrhythmia frequency and there was no correlation between animals with higher frequency of an arrhythmia type and the frequency of other arrythmias in the same animals. Clinical chemistry or hematology parameters were not different with or without telemetry devices. NHP with a transmural left ventricular pressure (LVP) catheter exhibited a greater incidence of VPBs and PJCs compared to telemetry animals without LVP. DISCUSSION: All species were similar with regards to the frequency of ventricular ectopic beats (26-46%) while the dog seemed to have more frequent junctional complexes and AVB compared to NHP and minipigs. Arrhythmia screening may be considered during pre-study evaluations, to exclude animals with abnormally high arrhythmia incidence.

[Effects of different long-term fertilization patterns on net carbon sink effect and economic benefits in double cropping rice paddy ecosystem in southern China]. / 长期不同施肥模式对南方双季稻田生态系统净碳汇效应和经济收益的影响.

There is close relationship between fertilizer managements and net carbon (C) sink effect, economic benefits in rice paddy ecosystem. Based on a long-term (35-year) field experiment, we analyzed the effects of different fertilization patterns on soil C sequestration rate, C density of topsoil, annual C balance, and economic benefits in the double cropping rice paddy in southern China. There were four fertilization treatments, chemical fertilizer alone (MF), rice straw and chemical fertilizer (RF), 30% organic manure and 70% chemical fertilizer (OM), and without any fertilizer input as a control (CK). The results showed that soil C pool in the double cropping rice paddy field under different fertilization treatments changed from 216.02 to 866.74 kg·hm-2·a-1, and soil C pool under OM treatment were significantly higher than that of MF, RF and CK. The soil C sequestration rates in the double cropping rice paddy field under different fertilization treatments ranged from 51.5 to 650.7 kg·hm-2·a-1, and that of C density of topsoil was from 55.64 to 78.42 t·hm-2. The order of soil C sequestration rates and C density of topsoil was OM>RF>MF>CK. The change range of C adsorption in the double cropping rice paddy field ecosystem was from 4.42 to 9.32 t C·hm-2·a-1, with an order of OM>RF>MF>CK. Compared with the MF treatment, soil net C sink under OM and RF treatments increased by 27.6% and 13.6%, respectively. The change range of C cost material input ranged from 1.49 to 2.17 t C·hm-2·a-1, and that of annual economic benefits was from 1.30×103 to 7.83×103 yuan·hm-2·a-1 with an order of RF>OM>MF>CK. The net income of economic benefits of OM, RF and MF treatments were significantly higher than that of CK. Generally, soil C sequestration rate, C sink effect and annual economic benefits were increased by the long-term application of organic manure and rice straw returning together with chemical fertilizer, which could increase soil organic carbon storage in the double cropping rice paddy field of southern China.

Cholesterol Metabolic Markers for Differential Evaluation of Patients with Hyperlipidemia and Familial Hypercholesterolemia.

The cholesterol metabolism in humans can be indirectly reflected by measuring cholesterol metabolism marker levels. We aimed to investigate the association of cholesterol homeostasis markers on standard lipid profiling components in familial hypercholesteremia and hyperlipidemia patients. A total of 69 hyperlipidemia patients, 25 familial hypercholesteremia (FHC) patients, and 64 healthy controls were enrolled in this study. We performed routine testing of blood lipid water. Gas chromatography was used to determine the changes in the concentration of cholesterol synthesis (squalene, desmosterol, and lathosterol) and absorption markers (campesterol, sitosterol, and stigmasterol) in the blood. Baseline hyperlipidemia patients displayed significantly higher total cholesterol (TC), triglyceride (TG), and low-density lipoprotein cholesterol (LDL-C) levels in comparison to the control group, which was reflected in the increased levels of squalene, desmosterol, campesterol, and sitosterol observed (P < 0.05) in the hyperlipidemia patients. The desmosterol, lathosterol, campesterol, stigmasterol, and sitosterol were statistically different in the FHC group than the hyperlipidemic group (P < 0.05). The proportions of squalene/cholesterol, lathosterol/cholesterol, stigmasterol/cholesterol, and sitosterol/cholesterol in the FHC group were lower than those in the hyperlipidemic group; only desmosterol/cholesterol was higher than that in the hyperlipidemic group. Correlation studies between lipid metabolic factors showed that the proportion of moderate and strong correlations was much higher in the FHC group than in the other two groups (76.92% vs. 32.50% and 31.25%). Logistic regression analysis showed that the concentrations of glucose, LDL-C, lactosterol, and sitosterol were all independent risk factors for developing hyperlipidemia. This result was further confirmed by the ROC curve. These results indicated that the study of cholesterol synthesis and decomposition markers can serve as a reference index for related diseases caused by changes in its concentration.

[Impacts of long-term different fertilization managements on soil acid hydrolysable organic nitrogen fractions in double-cropping rice field]. / é•¿æœŸä¸åŒæ–½è‚¥æ¨¡å¼å¯¹åŒå­£ç¨»ç”°åœŸå£¤é ¸è§£æœ‰æœºæ°®ç»„åˆ†çš„å½±å“.

To clarify the impacts of long-term different fertilization modes on the soil acid hydrolysable organic nitrogen and its components in the double-cropping rice field of southern China, a long-term (36-year) location field experiment was used as a platform to systematically analyze the variations of soil acid hydrolysable organic nitrogen and its components (amino acid nitrogen, amino sugar nitrogen, ammonium nitrogen, unidentified hydrolysable nitrogen) at 0-10 cm and 10-20 cm soil layers under four fertilization treatments, including chemical fertilizer alone (CF), rice straw and chemical fertilizer (RF), 30% organic manure and 70% chemical fertilizer (OM), and control (CK). The relationships of soil acid hydrolysable organic nitrogen content and soil total nitrogen, soil organic carbon content were also analyzed. The results showed that compared with CK, OM and RF treatments significantly increased the contents of total nitrogen, available nitrogen, and organic carbon at both soil layers. The contents of soil acid hydrolysable organic nitrogen at 0-10 cm and 10-20 cm layers of OM, RF and CF treatments were 10.7%-42.6% and 12.2% -51.5% higher than that of CK, respectively. Compared with CF and CK treatments, OM and RF treatments significantly increased the contents of amino acid nitrogen, ammonium nitrogen, unidentified hydrolysable nitrogen and amino sugar nitrogen contents atboth soil layers. The soil acid hydrolysable organic nitrogen and non-hydrolysable nitrogen contents at 0-10 cm and 10-20 cm paddy soils under different fertilization treatments decreased in an order of OM>RF>CF>CK. The contents of soil amino acid nitrogen, ammonium nitrogen, amino sugar nitrogen and non-hydrolysable nitrogen at 0-10 cm soil layer of each fertilization treatment were higher than those at 10-20 cm soil layer. In addition, the contents of soil acid hydrolyzed organic nitrogen were positively correated with the contents of soil total nitrogen and soil organic carbon. In conclusion, RF and OM treatments were beneficial to increase organic nitrogen content at 0-10 and 10-20 cm soil layers of double cropping paddy fields, with postive consequences on nitrogen supply capacity and soil fertility.

Effects of long-term fertilizer management on soil labile organic carbon fractions and hydrolytic enzyme activity under a double-cropping rice system of southern China.

Fertilization is an effective way to improve soil quality, increase soil fertility and soil microbial diversity in paddy soil. To explore the changes of soil labile organic carbon (C) fractions and hydrolytic enzyme activity after 34 years fertilization treatments in a field experiment in double-cropping rice system of southern China. There were four treatments, including chemical fertilizer alone (MF), rice residue and chemical fertilizer (RF), 30% organic matter and 70% chemical fertilizer (OM), and the control without fertilizer input (CK). We measured soil organic carbon (SOC) content, soil labile organic C fractions, SOC related hydrolytic enzyme activity, correlation coefficients of soil enzyme activity with SOC content and its labile organic C fractions. The results showed that MF, RF and OM increased SOC content by 4.5%, 22.4% and 53.5%, respectively. Compared with MF and CK, RF and OM increased soil labile organic C fractions [cumulative C mineralization (Cmin), permanganate oxidizable C (KMnO4-C), particulate organic C (POC), dissolved organic C (DOC), light fraction organic C (LFOC), microbial biomass C (MBC)] and the proportion of each labile organic C fractions to total organic C. The contents of Cmin, KMnO4-C, POC, DOC, LFOC and MBC under OM treatment were 3.5, 3.1, 3.7, 1.9, 1.2 and 1.9 times higher than CK treatment, respectively. The proportion of labile organic C fractions to total organic C of RF and OM treatments was significantly higher than that in CK. The order of soil hydrolytic enzyme activity [α-glucosidase (αG), ß-glucosidase (ßG), ß-xylosidase (ßX), cellobiohydrolase (GBH), and N-acetyl-ß-glucosaminidase (NAG)] was OM>RF>MF>CK. The soil hydrolytic enzyme activity under OM treatment increased by 111.8%, 14.1%, 127.3%, 285.6% and 91.4% compared with CK, respectively. Furthermore, RF and OM treatments were beneficial to soil peroxidase (POD) activity. MF treatment was beneficial to soil polyphenol oxidase (PPO) activity. There was a significant positive correlation between soil hydrolytic enzyme activity and SOC content and its labile organic C fractions. In conclusion, the combined application of organic manure, rice straw returning and chemical fertilizer is an effective method to improve soil labile organic C fractions and hydrolytic enzyme activity in a double-cropping rice paddy field of southern China.

Respiratory Rates Derived From Arterial Blood Pressure Waveforms in Telemetered Dogs.

The objective of this study was to extract low frequency respiratory "artifacts" from a standard arterial blood pressure (ABP) waveform to simultaneously derive reliable breathing rates (BR). Arterial blood pressure derived BR values were characterized against respiratory rates simultaneously obtained from the Respiratory Inductive Plethysmography (RIP) system (EMKA). Reference compounds were introduced to evaluate responsiveness of the derived measures to respiratory depressants and stimulants. Male beagle dogs (n = 3) were instrumented with minimally invasive telemetry devices for measurements of ABP and heart rate. The RIP system was utilized simultaneously to collect respiratory rate, tidal volume, and minute volume of each animal following pharmacological challenges. Early results revealed the derived BR's from ABP waveforms did not correlate well with those measured from the RIP system. Post study X-ray visualization revealed suboptimal catheter positioning, causing poor concordance of BR tallied from the ABP waveforms. Follow-up evaluations were conducted using additional animals instrumented with the ABP catheter tip placement advanced proximal to the thoracic diaphragm. Preliminary data from this subset of animals significantly improved the correlation of BR derived from ABP and respiratory rates recorded by the RIP. This proof of concept investigation was intended to evaluate an algorithm designed to extract additional data from routine cardiac waveforms. We clearly demonstrated that with optimal blood pressure catheter placement and acquisition algorithm, a reliable breathing rate can also be extracted from safety studies without the need for additional studies/animals to capture those respiratory end points.

Echocardiographic and hemodynamic indices of myocardial contractility simultaneously evaluated in telemetered beagle dogs: A HESI-sponsored cross-company evaluation.

INTRODUCTION: Alterations in cardiac contractility can have significant clinical implications, highlighting the need for early detection of potential liabilities. Pre-clinical methods to assess contractility are typically invasive and their translation to human measures of cardiac function are not well defined. Clinically, cardiac function is most often measured non-invasively using echocardiography. The objective of these studies was to introduce echocardiography into standard large animal cardiovascular safety pharmacology studies and determine the feasibility of this combination. METHODS: A consortia of laboratories combined their data sets for evaluation. At each site, telemetered beagle dogs, in a 4 × 4 Latin square crossover study design (n = 4), were administered either pimobendan (positive inotrope) or atenolol (negative inotrope) orally at clinically relevant dose levels. Standard telemetry parameters were collected (heart rate, mean arterial blood pressure, etc.) continuously over 24 h, as well as derived contractility endpoints: QA interval and LV +dP/dtmax. At Tmax, echocardiography was performed in conscious dogs with minimal restraint to collect contractility parameters: ejection fraction (EF) and fractional shortening (FS). RESULTS: Correlations between telemetry and echo contractility endpoints showed that, in general, a change in LV +dP/dtmax of 1000 mmHg/s translates to a 5.2% change in EF and a 4.2% change in FS. Poor correlations were shown between QA interval derived simultaneously, to both EF and FS. DISCUSSION: Comparing data from telemetry-only groups to those that included echocardiography collections showed no effect in the ability to interpret test article-related effects, providing the foundation for the inclusion of echocardiography without compromising standard telemetry data quality.

[Microbial Carbon Source Metabolic Profile in Rice Rhizosphere and Non-rhizosphere Soils with Different Long-term Fertilization Management].

Rhizosphere and non-rhizosphere soil samples under different long-term fertilization treatments including control without fertilizer (CK), chemical fertilization alone (NPK), rice residues combined with NPK (NPKS), 30% manure plus 70% chemical fertilizers (LOM), and 60% manure plus 40% chemical fertilizers (HOM) were collected from a paddy field in a red soil hilly area in Ningxiang City, Hunan Province, China. The characteristics of microbial carbon utilization in the soils were studied. Results of 18O-H2O tracer analysis showed that both soil microbial biomass carbon content (MBC) and microbial growth rate (CGrowth) were highest in the HOM treatment, whereas they were lowest in CK. In the rhizosphere soil, the highest basal respiration was observed in HOM, and the lowest values were in CK and NPK. Microbial carbon utilization efficiency (CUE) was highest in NPK but lowest in the LOM and HOM treatments. In non-rhizosphere soil, no significant differences between basal respiration and CUE were observed among the fertilization treatments. Results from MicroRespTM showed that the ability of microorganisms to metabolize exogenous carbon sources was higher in non-rhizosphere soil than in rhizosphere soil. The application of organic materials (rice residues or manure) increased the microbial metabolic rate of carboxylic acids, amino acids, and carbohydrates in the order carboxylic acids > amino acids and carbohydrates > complex compounds. Redundancy analysis of the microbial metabolism patterns of various carbon substrates showed that:① CK was well separated from the fertilization treatments; ② NPK was grouped with NPKS, whereas LOM and HOM were grouped together and were separate from NPK and NPKS. This indicates that the fertilization treatments changed the microbial carbon metabolism patterns. The above-mentioned results indicated that the fertilization treatments did not affect microbial CUE and basal respiration. However, exogenous carbon source input (such as root exudates) and the application of organic materials can increase microbial basal respiration, and thus, reduce microbial CUE.

[Effects of different planting patterns on the apparent balance of soil nutrients and nitrogen production efficiency in paddy soil]. / ç¨»ç”°ä¸åŒç§æ¤æ¨¡å¼å¯¹åœŸå£¤å »åˆ†è¡¨è§‚å¹³è¡¡å’Œæ°®ç´ ç”Ÿäº§æ•ˆçŽ‡çš„å½±å“.

To investigate the effects of different planting patterns on soil nutrient contents, rice yield, nitrogen production efficiency, and apparent nutrient balance, a long-term field experiment was conducted in Changsha, Hunan, China. Four planting patterns were examined, including winter fallow-double cropping rice (CK), ryegrass-double cropping rice (Ry-R-R), Chinese milk vetch-double cropping rice (Mv-R-R), rape-double cropping rice (Ra-R-R). Compared with CK, soil total nitrogen (N) and available N were significantly higher under Ry-R-R, Mv-R-R and Ra-R-R. However, soil available potassium (K) content in CK was significantly higher than that in the other treatments. In addition, the N partial factor productivity in the late rice cropping was significantly higher under Ry-R-R, Mv-R-R and Ra-R-R. The N accumulation in panicles, stems and leaves of late rice under Ry-R-R, Mv-R-R and Ra-R-R were significantly higher than that in CK. Furthermore, the rice yield in Ry-R-R and Mv-R-R were significantly higher in comparison to CK. A significant positive correlation between grain yield and soil N content was observed. Compared with CK, Ry-R-R and Mv-R-R significantly increased the surplus of soil N and phosphorus content, but Ry-R-R and Ra-R-R increased the surplus of K content. Overall, winter crop-double rice cropping system could improve soil nutrient contents and N production efficiency, alleviate K deficiency, which would be beneficial to soil nutrient balance of paddy soils. Given the high surplus of N in paddy soils, N fertilizers should be reduced.

[Effects of different manure nitrogen input ratio on rhizosphere soil microbial biomass carbon, nitrogen and microbial quotient in double-cropping rice field]. / æœ‰æœºè‚¥æ°®æŠ•å ¥æ¯”ä¾‹å¯¹åŒå­£ç¨»ç”°æ ¹é™ åœŸå£¤å¾®ç”Ÿç‰©ç”Ÿç‰©é‡ç¢³ã€æ°®å’Œå¾®ç”Ÿç‰©ç†µçš„å½±å“.

To explore the characteristics of rhizosphere soil microorganisms in paddy fields with different manure nitrogen (N) input ratios at different growth stages of early and late rice in double-cropping rice system, a field experiment was conducted with five different treatments: 1) 100% N of chemical fertilizer (M1), 2) 30% N of organic matter and 70% N of chemical fertilizer (M2), 3) 50% N of organic matter and 50% N of chemical fertilizer (M3), 4) 100% N of organic matter (M4), and 5) no N fertilizer input as a control (M0). The rhizosphere soil microbial biomass carbon (MBC), microbial biomass nitrogen (MBN), and microbial quotient (SQ) of the paddy fields were measured using the fumigation-extraction and chemical analysis methods. The results showed that the rhizosphere MBC, MBN, and SQ of the paddy fields at main different growth stages of early and late rice were increased by fertilization, which increased first and then decreased with the development of rice growth period, peaked at the heading stage, and reached the minimum value at the maturity stage. The effects of different fertilization treatments were in order of M4＞M3＞M2＞M1＞M0, with no significant difference among M2, M3 and M4, but being significantly higher than M0. Therefore, the application of organic matter, and combined application of chemical fertilizer with organic matter could significantly increase the rhizosphere MBC, MBN, and SQ of the paddy fields at early and late rice growth period, while chemical fertilizer alone had little effect.

[Characteristics of amino sugar accumulation in rhizosphere and non rhizosphere soil of rice under different fertilization treatments]. / ä¸åŒæ–½è‚¥å¤„ç†ä¸‹æ°´ç¨»æ ¹é™ å’Œéžæ ¹é™ åœŸå£¤ä¸­æ°¨åŸºç³–ç§¯ç´¯ç‰¹å¾.

Soil samples were collected from paddy ecosystem under five long-term fertilization treatments, including control without fertilizer (CK), chemical fertilization alone (NPK), rice residue combined with NPK (NPKS), 30% manure plus 70% chemical fertilizer (LOM), and 60% manure plus 40% chemical fertilizer (HOM) in Ningxiang City, Hunan Province. The cha-racteristics of amino sugars accumulation in the rhizosphere and non-rhizosphere soils at rice tillering stage were analyzed. Results showed that the contents of soil organic carbon, total amino sugars and three amino monosaccharides (muramic acid, glucosamine and galactosamine) with long-term application of organic materials (rice residue or manure) were significantly higher compared with CK and NPK. The inconsistent accumulation trends of the three amino monosaccharides under different fertilization treatments indicated that different responses of microbial groups to various fertilization treatments. The content of total amino sugars was not significantly different between the rhizosphere soil and the non-rhizosphere soil, probably because the agricultural operations such as plowing could homogenize paddy soils. The contribution of amino sugar derived carbon to soil organic carbon ranged from 24.0 to 28.3 mg·g-1, which was highest in NPKS, and lowest in HOM and CK. The ratio of fungal to bacterial residues (fungal glucosamine/muramic acid) ranged from 24.4 to 36.6, indicating that fungi dominated the degradation and transformation of organic matter in all the soils. Compared with that under NPK and CK, the participation of organic matter transformation from fungi under NPKS treatment was increased, whereas the bacteria involved in organic matter transformation under HOM treatment was enhanced.

[Effects of Long-term Fertilization Regimes on Microbial Biomass, Community Structure and Activity in a Paddy Soil].

Four paddy soils were collected in Ningxiang County, Hunan province. These used with different long-term fertilization regimes, including a control without fertilizer (CK), chemical fertilization with nitrogen, phosphate, and kalium (NPK), straw fertilization combined with NPK (ST), and manure fertilization combinedwith NPK (OM). Phospholipid fatty acid (PLFA) technology and MicrorespTM method were used to study the effect of long-term fertilization on soil microorganism abundance, community structure, and activity. Results showed that the abundance of bacteria, fungi, gram-negative (G-) bacteria, and gram-positive (G+) bacteria in the soil from the OM treatment was generally higher than for the other treatments; these levels were lower in the ST and NPK treatments and lowest in the CK treatment. The principal components analysis (PCA) of PLFA showed that the community structure of microorganisms in NPK, ST, and OM treatments was altered in comparison with that in CK, especially in the case of the ST and OM treatments. MicroRespTM results revealed that compared to the CK treatment (1.28 µg·h-1), soil microorganisms in the OM treatment had the highest average utilization rate of multiple carbon sources (1.81 µg·h-1), followed by ST (1.19 µg·h-1), CK (1.28 µg·h-1), and NPK (0.95 µg·h-1). Furthermore, different long-term fertilization regimes resulted in distinct carbon source preferences for the soil microorganisms and revealed a significant alteration in the microbial community. Conclusively, long-term fertilizer with straw or manure changes the microbial community and is a benefit for improving the biomass and activity of microorganism in rice paddy soils.

Integrated TK-TD modeling for drug-induced concurrent tachycardia and QT changes in beagle dogs.

Drug-induced cardiotoxicity, including tachycardia and QT prolongation, remains a major safety concern that needs to be identified and its risk mitigated in early stages of drug development. In the present study, an integrated toxicokinetic-toxicodynamic (TK-TD) modeling approach within a nonlinear mixed-effect modeling framework is applied to investigate concurrent abnormal heart rate and QT changes in three beagle dogs, using a Novartis internal compound (NVS001) as the case example. By accounting for saturable drug absorption, circadian rhythms, drug-effect tolerance, and nonlinear rate-dependency of QT interval, the dynamic TK-TD model captures the experimentally observed drug effects on heart rate and QT interval across a wide dosing range of NVS001 in beagle dogs. Further analyses reveal that the NVS001-induced QT prolongation observed in the low-dose groups is potentially caused by direct drug inhibition on the hERG channel, while the apparent QT shortening in the high-dose groups may be due to strong rate-dependency of QT at high heart rates. This study also suggests that the TK-TD model can be used to identify direct drug effects on the non-rate-dependent QT component by dissociating QT changes from tachycardia and deriving a new QT correction method. The integrated TK-TD model presented here may serve as a novel quantitative framework for evaluating drug-induced concurrent changes in heart rate and QT to potentially facilitate preclinical and clinical safety studies.

[Effects of different winter cover crops on soil organic carbon in a double cropping rice paddy field.] / 不同冬季覆盖作物对双季稻田土壤有机碳的影响.

In a double cropping rice field experiment, effects of five winter cover crops on the total organic carbon (TOC), active organic carbon (AOC), carbon pool management index (CPMI) and organic carbon storage were studied in three soil layers (0-5, 5-10 and 10-20 cm).Winter cover crops of ryegrass (Ry), Chinese milk vetch (Mv), potato (Po), and rape (Ra) between two rice crops were compared with fallow as control (CK). The results showed that the TOC and AOC contents under Ry, Mv, Po and Ra treatments were higher than those of CK in all three la-yers. Meanwhile, the TOC and AOC contents in Po treatment were higher than those of other treatments. Compared with CK, the AOC, activity index (AI), carbon pool index (CPI) and CPMI in the soil were improved through the recycling of winter cover crops straw. The AOC, AI, CPI and CPMI in the studied layers increased in order of Po>Mv>Ry>Ra>CK. The results indicated that the recycling of winter cover crops straw promoted the storage of SOC in the 0-20 cm soil profile as compared with CK. The strongest effect of the winter cover crops on the SOC storage occurred in Mv treatment, followed by Mv and Po treatments, and the SOC storage increased with the increasing soil depth.

Evaluation of the effects of the water-soluble total flavonoids from Isodon lophanthoides var.gerardianus (Benth.) H. Hara on apoptosis in HepG2 cell: Investigation of the most relevant mechanisms.

ETHNOPHARMACOLOGICAL RELEVANCE: The water-soluble total flavonoids (WSTF) were extracted from Isodon lophanthoides var. gerardianus (Benth.) H. Hara, a common folk herbal medicine in China, which has been recorded by the "Chinese Pharmacopoeia" in 2015 and used for prevention and clinical treatment of common diseases of liver and gall for many years. OBJECTIVE OF THE STUDY: The aim of this study is to evaluate the effects of WSTF on apoptosis in HepG2 cell and investigate the relevant mechanisms underlying. MATERIALS AND METHODS: Cytotoxicity was evaluated in HepG2 cells (human hepatoma cell lines) using MTT assay. The influence of the WSTF on the intracellular reactive oxygen species (iROS) and the mitochondrial membrane potential were also determinated. We used flow cytometry analysis to detect the effects of WSTF on apoptosis, cell cycle. Then we applied RT-PCR for genetic expression of main effectors and western blot analysis for activation of main effectors involved in the potential apoptosis signaling pathways. RESULTS: WSTF inhibited cell growth in HepG2 cells. Moreover, WSTF stimulates to increase amount of iROS, mitochondrial membrane potential, and the apoptotic relevant factors (cytochrome c, caspase-3) in HepG2 cells. WSTF could significantly induce apoptosis through downregulating apoptosis-antagonizing protein (Bcl-2, Survivin, mcl-1) and upregulating apoptosis-promoting proteins (Bax) and cell cycle G0/G1 arrest in HepG2 cells. CONCLUSION: The results indicate that WSTF induces cell apoptosis through mitochondrial pathway in the HepG2 cells. Therefore, these studies suggest that WSTF could be used as a chemotherapeutic agent to treat hepatoma.

Translational assessment of cardiac contractility by echocardiography in the telemetered rat.

INTRODUCTION: Cardiac contractility was evaluated using standard inotropic agents in rats. We compared indices of cardiac contractility, i.e. LV dP/dt max from telemetry while simultaneously collecting EF (ejection fraction) and FS (fractional shortening) measures from echocardiography. METHODS: Male Wistar rats were instrumented with telemetry devices for measurements of blood pressure and left ventricular pressure. Milrinone (PDE III inhibitor) and verapamil (L-type calcium channel blocker) at doses of 0, 3, 10, and 30 mg/kg were administered orally using a 4 × 4 Latin square crossover study design. Telemetry data were recorded at predose and continuously for 24h post-dose. Echocardiographic evaluations were conducted once at predose and at 1 and 2h after milrinone or verapamil administration, respectively. During the recording of echocardiograms, telemetry data were collected simultaneously. Blood samples were also collected to confirm plasma drug exposure. RESULTS: As expected, milrinone increased LV dP/dt max, EF and FS while verapamil decreased LV dP/dt max, EF and FS. Linear regression analysis showed a positive correlation between LV dP/dt max and EF or FS (P<0.001) with both test agents. A change in LV dP/dt max of 1000 mmHg/s was found to correspond with a change in EF and FS of 13 and 16%, respectively, in the telemetered rat. DISCUSSION: The correlation between contractility indices assessed by telemetry and echocardiographic methods in rat models has not received much attention to date. Our results with two reference compounds demonstrate that both methods are sensitive to alterations in contractility induced by inotropic agents administered to rats. The high degree of correlation between changes in LV dP/dt max and EF or FS in the rat enables a translational-element of clinical relevance following changes in contractility indices when measured with telemetry devices in preclinical studies.

[Determination of six C-Glycoside flavones and antitumor activity of water-soluble total flavonoids from Isodon lophanthoides var. gerardianus].

This research established an HPLC method for determination of six C-Glycoside flavones of warer-soluble total flavonoids from Isodon lophanthoides var. gerardianus (Benth.) H. Hara, and studied the antitumor activity of the warer-soluble total flavonoids. The HPLC system consisted of Kromasil 100-5 C18 (4.6 mm x 250 mm, 5 microm) column and a solution system of methanol, acetonitrile and 0.5% formic acid gradient elution at a flow rate of 0. 8 mL x min(-1) and the wavelength of detector was at 334 nm. The column temperature was 25 degrees C. The antitumor activity of water-soluble flavonoids was assayed using HepG2 cell as the tested cell. The linear ranges of vicenin II, vicenin III, isoschaftoside, schaftoside, vitexin, 6, 8-di-C-a-L-arabinosylapigenin were 0.25-2.53, 0.12-1.20, 0.37-3.69, 0.16-1.63, 0.19-1.92, 0.14-1.42 microg, respectively. The average recoveries (n = 6) were 99.6% (RSD 0.87%), 100.2% (RSD 2.0%), 99.6% (RSD 1.8%), 97.9% (RSD 1.5%), 98.8% (RSD 1.2%), 98.6% (RSD 1.2%), respectively. After exposure in 24, 48, 72 h, the total flavonoids showed inhibitory effect on the proliferation of HepG2 cells with IC50 as the evaluation index, the IC50 values of 1.89, 1.71, 1.51 g x L(-1), respectively. The method is quick, simple and accurate with good re- producibility, and can be used for determination of vicenin II, vicenin III, isoschaftoside, schaftoside, vitexin, 6, 8-di-C-a-L-arabino- sylapigenin in the warer-soluble total flavonoids from L lophanthoides var. gerardianus. The warer-soluble total flavonoids from L lophanthoides have inhibitory effect on the proliferation of HepG2 cells.

Englerin A Agonizes the TRPC4/C5 Cation Channels to Inhibit Tumor Cell Line Proliferation.

Englerin A is a structurally unique natural product reported to selectively inhibit growth of renal cell carcinoma cell lines. A large scale phenotypic cell profiling experiment (CLiP) of englerin A on ¬over 500 well characterized cancer cell lines showed that englerin A inhibits growth of a subset of tumor cell lines from many lineages, not just renal cell carcinomas. Expression of the TRPC4 cation channel was the cell line feature that best correlated with sensitivity to englerin A, suggesting the hypothesis that TRPC4 is the efficacy target for englerin A. Genetic experiments demonstrate that TRPC4 expression is both necessary and sufficient for englerin A induced growth inhibition. Englerin A induces calcium influx and membrane depolarization in cells expressing high levels of TRPC4 or its close ortholog TRPC5. Electrophysiology experiments confirmed that englerin A is a TRPC4 agonist. Both the englerin A induced current and the englerin A induced growth inhibition can be blocked by the TRPC4/C5 inhibitor ML204. These experiments confirm that activation of TRPC4/C5 channels inhibits tumor cell line proliferation and confirms the TRPC4 target hypothesis generated by the cell line profiling. In selectivity assays englerin A weakly inhibits TRPA1, TRPV3/V4, and TRPM8 which suggests that englerin A may bind a common feature of TRP ion channels. In vivo experiments show that englerin A is lethal in rodents near doses needed to activate the TRPC4 channel. This toxicity suggests that englerin A itself is probably unsuitable for further drug development. However, since englerin A can be synthesized in the laboratory, it may be a useful chemical starting point to identify novel modulators of other TRP family channels.

[Effects of long-term tillage and rice straw returning on soil nutrient pools and Cd concentration].

The objective of this study was to assess the effects of tillage and straw returning on soil nutrient and its pools, and soil Cd concentration, and to identify the strategies for rational tillage and remediation of Cd contaminated paddy fields. The experiment was established with no-tillage with straw retention (NTS) , rotary tillage with straw incorporation (RTS) , conventional plow tillage with straw incorporation (CTS), conventional plow tillage with straw removed ( CT) from 2005 to 2013. The results indicated that tillage and rice straw retention had a great impact on soil properties at 0-10 cm soil depth. The soil aeration, and concentrations of soil nutrient and soil Cd increased under CTS, CT, and RTS. Due to the shallow plow layers, soil nutrient pools and the Cd concentration in rice shoot decreased in long-term tilled soil. Under long-term no-tillage, the soil bulk, soil nutrient pools and Cd concentration in rice shoot increased, but concentrations of soil nutrients decreased. In addition, rice straw returning significantly increased the soil nutrient concentrations, cation exchange capacity, depth of plow layer, and soil nutrient pools. However, the Cd in the rice straw was also returned to the soil by rice straw returning, which would not benefit the remediation of soil Cd. Therefore, it is necessary to improve tillage and straw retention practices due to the disadvantages of long-term continuous single tillage method and rice straw returning practices. Some recommended managements (e.g., rotational tillage or subsoiling, reducing straw returning amount, and rotational straw returning) could be good options in enhancing soil fertility and remedying soil pollution.