A covalently bound photoisomerizable agonist. Comparison with reversibly bound agonists at electrophorus electroplaques

After disulphide bonds are reduced with dithiothreitol, trans-3- (α-bromomethyl)-3’-[α- (trimethylammonium)methyl]azobenzene (trans-QBr) alkylates a sulfhydryl group on receptors. The membrane conductance induced by this “tethered agonist” shares many properties with that induced by reversible agonists. Equilibrium conductance increases as the membrane potential is made more negative; the voltage sensitivity resembles that seen with 50 [mu]M carbachol. Voltage- jump relaxations follow an exponential time-course; the rate constants are about twice as large as those seen with 50 μM carbachol and have the same voltage and temperature sensitivity. With reversible agonists, the rate of channel opening increases with the frequency of agonist-receptor collisions: with tethered trans-Qbr, this rate depends only on intramolecular events. In comparison to the conductance induced by reversible agonists, the QBr-induced conductance is at least 10-fold less sensitive to competitive blockade by tubocurarine and roughly as sensitive to “open-channel blockade” bu QX-222. Light-flash experiments with tethered QBr resemble those with the reversible photoisomerizable agonist, 3,3’,bis-[α-(trimethylammonium)methyl]azobenzene (Bis-Q): the conductance is increased by cis {arrow} trans photoisomerizations and decreased by trans {arrow} cis photoisomerizations. As with Bis-Q, ligh-flash relaxations have the same rate constant as voltage-jump relaxations. Receptors with tethered trans isomer. By comparing the agonist-induced conductance with the cis/tans ratio, we conclude that each channel’s activation is determined by the configuration of a single tethered QBr molecule. The QBr-induced conductance shows slow decreases (time constant, several hundred milliseconds), which can be partially reversed by flashes. The similarities suggest that the same rate-limiting step governs the opening and closing of channels for both reversible and tethered agonists. Therefore, this step is probably not the initial encounter between agonist and receptor molecules.     

5种相思树和尾巨桉人工林土壤养分和酶活性特征

为揭示固氮树种土壤养分转化的酶学机制,对固氮树种[厚荚相思(Acacia crassicarpa)、黑木相思(A. melanoxylon)、卷荚相思(A.cincinnata)、大叶相思(A.auriculiformis)和马占相思(A.mangium)]及非固氮树种尾巨桉(Eucalyptusurophylla×E.grandis)人工林的土壤养分含量、酶活性及其相关性进行研究。结果表明,相思林40～60cm土层的pH均高于尾巨桉林;5种相思林土壤各土层的TP、TK含量均低于尾巨桉林,而20～40 cm土层的TC、TN含量均高于尾巨桉林,黑木相思林和马占相思林各土层的有效养分均显著高于尾巨桉林(P0.05)。0～10 cm土层中,相思林的土壤酸性磷酸酶和纤维素酶活性均高于尾巨桉林,大叶相思林的土壤脲酶、蔗糖酶、纤维素酶和芳基硫酸酯酶活性显著高于尾巨桉林(P0.05),卷荚相思林的土壤脲酶、纤维素酶、几丁质酶和淀粉酶活性显著高于尾巨桉林(P0.05)。相关分析结果表明,土壤脲酶、蔗糖酶和几丁质酶活性与AP显著负相关(P0.05),蔗糖酶和纤维素酶活性与NH4+-N显著负相关(P 0.05),脲酶、纤维素酶、芳基硫酸酯酶与土壤TK显著负相关(P0.05),几丁质酶活性与TN含量呈显著正相关(P0.05),土壤淀粉酶活性与NH4+-N呈显著正相关(P 0.05),过氧化氢酶活性与土壤TK含量呈显著正相关。可见,与尾巨桉人工林相比,在我国南方退化山地引种相思树可提高土壤关键酶的活性,对土壤有效养分具有明显改良作用,有利于退化地土壤的生态修复及人工林长期生产力的维持。     

Citrullus lanatus ‘sentinel’ (watermelon) extract reduces atherosclerosis in LDL receptor-deficient mice

Watermelon (Citrullus lanatus or C. lanatus) has many potentially bioactive compounds including citrulline, which may influence atherosclerosis. In this study, we determined the effects of C. lanatus, provided as an extract of the cultivar ‘sentinel,’ on hypercholesterolemia-induced atherosclerosis in mice. Male low-density lipoprotein receptor-deficient mice at 8 weeks old were given either C. lanatus ‘sentinel’ extract (2% vol/vol; n= 10) or a mixture of matching carbohydrates (2% vol/vol; n= 8) as the control in drinking water while being fed a saturated fat-enriched diet for 12 weeks ad libitum. Mice consuming C. lanatus ‘sentinel’ extract had significantly increased plasma citrulline concentrations. Systolic blood pressure was comparable between the two groups. Consumption of C. lanatus ‘sentinel’ extract led to lower body weight and fat mass without influencing lean mass. There were no differences in food and water intake and in urine output between the two groups. C. lanatus ‘sentinel’ extract administration decreased plasma cholesterol concentrations that were attributed to reductions of intermediate-/low-density lipoprotein cholesterol. Plasma concentrations of monocyte chemoattractant protein-1 and interferon-gamma were decreased and those of interleukin-10 were increased in mice consuming C. lanatus ‘sentinel’ extract. Intake of C. lanatus ‘sentinel’ extract resulted in reductions of atherosclerosis in both aortic arch and thoracic regions. In conclusion, consumption of C. lanatus ‘sentinel’ extract led to reduced body weight gain, decreased plasma cholesterol concentrations, improved homeostasis of pro- and anti-inflammatory cytokines, and attenuated development of atherosclerosis without affecting systolic blood pressure in hypercholesterolemic mice.     

Revealing the True Incidence of Pandemic A(H1N1)pdm09 Influenza in Finland during the First Two Seasons — An Analysis Based on a Dynamic Transmission Model

The threat of the new pandemic influenza A(H1N1)pdm09 imposed a heavy burden on the public health system in Finland in 2009-2010. An extensive vaccination campaign was set up in the middle of the first pandemic season. However, the true number of infected individuals remains uncertain as the surveillance missed a large portion of mild infections. We constructed a transmission model to simulate the spread of influenza in the Finnish population. We used the model to analyse the two first years (2009-2011) of A(H1N1)pdm09 in Finland. Using data from the national surveillance of influenza and data on close person-to-person (social) contacts in the population, we estimated that 6% (90% credible interval 5.1 – 6.7%) of the population was infected with A(H1N1)pdm09 in the first pandemic season (2009/2010) and an additional 3% (2.5 – 3.5%) in the second season (2010/2011). Vaccination had a substantial impact in mitigating the second season. The dynamic approach allowed us to discover how the proportion of detected cases changed over the course of the epidemic. The role of time-varying reproduction number, capturing the effects of weather and changes in behaviour, was important in shaping the epidemic.     

Predictive Values of N-Terminal Pro-B-Type Natriuretic Peptide and Cardiac Troponin I for Myocardial Fibrosis in Hypertrophic Obstructive Cardiomyopathy

Background

Both high-sensitivity cardiac troponin T and B-type natriuretic peptide are useful in detecting myocardial fibrosis, as determined by late gadolinium enhancement (LGE) cardiovascular magnetic resonance (CMR), in patients with non-obstructive hypertrophic cardiomyopathy. However, their values to predict myocardial fibrosis in hypertrophic obstructive cardiomyopathy (HOCM) remain unclear. We investigated the role of N-Terminal Pro-B-Type Natriuretic Peptide (NT-proBNP) and cardiac troponin I (cTnI) to identify LGE-CMR in patients with HOCM.

Methods

Peripheral concentrations of NT-proBNP and cTnI were determined in patients with HOCM (n = 163; age = 47.2 ± 10.8 years; 38.7% females). Contrast-enhanced CMR was performed to identify and quantify myocardial fibrosis.

Results

LGE was detected in 120 of 163 patients (73.6%). Patients with LGE had significantly higher levels of NT-proBNP and cTnI than those without LGE (1386.2 [904.6–2340.8] vs. 866.6 [707.2–1875.2] pmol/L, P = 0.003; 0.024 [0.010–0.049] vs. 0.010 [0.005–0.021] ng/ml, P <0.001, respectively). The extent of LGE was positively correlated with log cTnI (r = 0.371, P <0.001) and log NT-proBNP (r = 0.211, P = 0.007). On multivariable analysis, both log cTnI and maximum wall thickness (MWT) were independent predictors of the presence of LGE (OR = 3.193, P = 0.033; OR = 1.410, P < 0.001, respectively), whereas log NT-proBNP was not. According to the ROC curve analysis, combined measurements of MWT ≥21 mm and/or cTnI ≥0.025ng/ml indicated good diagnostic performance for the presence of LGE, with specificity of 95% or sensitivity of 88%.

Conclusions

Serum cTnI is an independent predictor useful for identifying myocardial fibrosis, while plasma NT-proBNP is only associated with myocardial fibrosis on univariate analysis. Combined measurements of serum cTnI with MWT further improve its value in detecting myocardial fibrosis in patients with HOCM.     

中国古老月季‘月月粉’FT/TFL1基因的鉴定与表达分析

该研究以中国古老月季品种‘月月粉’(Rosa chinensis‘Old Blush’)为试材,根据拟南芥和草莓FT/TFL1基因家族成员的保守结构域设计引物,对‘月月粉’基因组DNA进行克隆和测序,结合数据库序列和双单倍体基因组信息进行序列比对,结果共获得了8条月季的FT/TFL1基因家族序列,且8条序列分别注释为2个FT基因(RcFT1和RcFT2)、1个BFT基因(RcBFT)、1个ATC基因(RcATC)、2个MFT基因(RcMFT1和RcMFT2)和2个PEBP基因(PEBP1和PEBP2);进化树分析结果将这8个基因分为3个亚组;结构分析显示8个基因的内含子为1～5个不等。qRT-PCR分析显示,RcFT1、RcPEBP、RcMFT1、RcMFT2和RcBFT在‘月月粉’的叶片中表达量最高,茎尖中次之;RcFT2在茎尖中的表达最高,叶片中次之;RcATC在茎尖中的表达量高于其他组织;除RcPEBP在根组织中有表达外,其他基因在根组织中几乎没有检测到。进一步表达分析显示,RcFT1和RcFT2在‘无刺光叶蔷薇’(R.wichuriana‘Basye’s Thornless')生殖生长期的叶片、茎尖中的表达量显著高于营养生长期。研究推测,具有连续开花性状的中国古老月季‘月月粉’可能具有多个开花时间(FT)基因位点,FT有可能可以作为月季生殖转换的标记基因。     

Rescue of odontogenesis in Dmp1-deficient mice by targeted re-expression of DMP1 reveals roles for DMP1 in early odontogenesis and dentin apposition in vivo

Dentin matrix protein 1 (DMP1) is expressed in both pulp and odontoblast cells and deletion of the Dmp1 gene leads to defects in odontogenesis and mineralization. The goals of this study were to examine how DMP1 controls dentin mineralization and odontogenesis in vivo. Fluorochrome labeling of dentin in Dmp1-null mice showed a diffuse labeling pattern with a 3-fold reduction in dentin appositional rate compared to controls. Deletion of DMP1 was also associated with abnormalities in the dentinal tubule system and delayed formation of the third molar. Unlike the mineralization defect in Vitamin D receptor-null mice, the mineralization defect in Dmp1-null mice was not rescued by a high calcium and phosphate diet, suggesting a different effect of DMP1 on mineralization. Re-expression of Dmp1 in early and late odontoblasts under control of the Col1a1 promoter rescued the defects in mineralization as well as the defects in the dentinal tubules and third molar development. In contrast, re-expression of Dmp1 in mature odontoblasts, using the Dspp promoter, produced only a partial rescue of the mineralization defects. These data suggest that DMP1 is a key regulator of odontoblast differentiation, formation of the dentin tubular system and mineralization and its expression is required in both early and late odontoblasts for normal odontogenesis to proceed.