Effect of diastolic pressure on MLC2v phosphorylation in the rat left ventricle

The effect of passive muscle stretch on the extent of MLC2v phosphorylation was investigated. We used an isolated rat heart preparation and controlled the passive pressure of the left ventricle (LV) at 0 or 15 mmHg. The hearts were flash frozen and the LV free wall was split into epicardial and the endocardial halves. The samples were solubilized using a novel method that minimizes changes in the phosphate content of MLC2v under non-denaturing conditions. The proteins were separated by urea glycerol PAGE and identified by mass spectrometry and Western blots. At 0 mmHg passive pressure, the extent of MLC2v phosphorylation of the epicardium (34.1+/-1.7%) was the same as that of the endocardium (35.3+/-3.4%). At 15 mmHg passive pressure, we found a significant increase in MLC2v phosphorylation in the epicardium (to 41.5+/-2.0%) and a significant reduction in the endocardium (to 24.2+/-1.2%), giving rise to a gradient in the extent of MLC2v phosphorylation from epicardium (high) to endocardium (low). These changes in MLC2v phosphorylation that take place in response to increased diastolic pressure are likely to impact on the calcium sensitivity of actomyosin interaction (with an increased sensitivity towards the epicardium) and may play a role in the Frank-Starling mechanism of the heart.     

Photothermal model of plant development

The development of plants depends on the photoperiod length, light intensity, temperature, and length of light day integral. The reaction of a plant to the day length or daily light integral can depend on both the range of studied light intensities and photoperiod. Based on the data concerning the effects of light and thermal integrals on the developmental rate of plants of different photoperiodic groups, a photothermal model of plant development was proposed. The model was used to calculate the lengths of optimal photoperiods and ranges of daily temperature gradients ensuring the highest developmental rate of some plants, such as soybean, wheat, cucumber, and barley.     

In the presence of ferritin, visible light induces lipid peroxidation of the porcine photoreceptor outer segment

We studied the synergistic effect of visible light and ferritin on the lipid peroxidation on a fraction of porcine photoreceptor outer segment (POS). Reaction mixtures containing the POS fraction and horse spleen ferritin were irradiated under white fluorescent light mainly at 17,000 lx or incubated under dark conditions at 37°C. The lipid peroxidation was evaluated by both the thiobarbituric acid method and the ferrous oxidation/xylenol orange method. The irradiation-induced lipid peroxidation was affected by some experimental factors such as the irradiation dose and acidity of the material. When the irradiation was stopped, the lipid peroxidation was also stopped; thereafter, the re-irradiation induced lipid peroxidation. Moreover, this lipid peroxidation was inhibited by desferrioxamine, an iron chelator, or by dimethylthiourea, a hydroxyl radical scavenger, suggesting that the lipid peroxidation involves hydroxyl radicals generated via the Fenton reaction by iron ion released from ferritin. The lipid peroxidation did not take place under dark conditions or in the absence of ferritin. This study suggested the possibility that the visible light-induced lipid peroxidation of the POS fraction in the presence of ferritin may participate in the etiology of human retinal degenerative diseases as the human retina is exposed to light for life.     

Protein phosphatase activity is necessary for myofibrillogenesis

During myofibrillogenesis, myosin light-chain kinase (MLCK) phosphorylates the regulatory light chain (RLC) of myosin II, enabling patterned assembly of myosin thick filaments. A protein phosphatase (PP) has been shown to mediate RLC dephosphorylation in adult smooth and striated muscle. A role for PP activity in regulating myofibrillogenesis during embryonic development, however, has not been investigated. Tautomycin (TM) was used to inhibit both PP1 and PP2A activities, whereas okadaic acid (OA) and fostriecin (FOS) were used to inhibit PP2A. TM affected both actin and myosin assembly at 5nM; the IC50 value was 20 and 8.5nM, respectively. In contrast, OA applied at 10 times above its reported Ki for PP2A caused no significant disruption. There was also no disruption when FOS was applied at a concentration 30 times above its reported Ki for PP2A. Thus, our results suggest a primary role for PP1 isoforms during myofibrillogenesis. Although rho kinase (RK) regulates PP activity in embryonic smooth and cardiac muscle, application of the RK inhibitor Y27632 did not affect actin or myosin assembly in skeletal myocytes. Collectively, our pharmacological results suggest that PP1 is involved in dynamic regulation of RLC phosphorylation. To specifically test involvement of the myosin-targeted isoform (PP1M), we used a morpholino antisense approach to knock down the myosin targeting (M) subunit of PP1. Embryos injected with morpholino targeted to the 110-kDa M targeting subunit had fewer somites, and myosin organization was significantly perturbed. The combined pharmacological and molecular results suggest a dynamic equilibrium between MLCK and PP1M activities is required for proper myofibrillogenesis.     

Comparison of a Salmonella typhimurium proteome defined by shotgun proteomics directly on an LTQ-FT and by proteome pre-fractionation on an LCQ-DUO.

Shotgun proteomics is rapidly becoming one of the most efficient and popular tools to examine protein expression in cells. Numerous laboratories now have a wide array of low- and high-performance mass spectrometry instrumentation necessary to complete proteome-wide projects. Often these laboratories have time and financial constraints that prohibit all projects from being conducted on high-performance state-of-the-art mass spectrometers. Here, we compare shotgun proteomic results using a direct 'lyse, digest and analyse' approach on a high-performance mass spectrometer (i.e. the LTQ-FT) with the results from a much lower-performance instrument (i.e. the LCQ-DUO) where, for the latter, various traditional protein pre-fractionation steps and gas-phase fractionation were used to increase the proteome coverage. Our results demonstrate that shotgun proteomic analyses conducted on the lower-performance LCQ-DUO mass spectrometer could adequately characterize a PhoP constitutive strain of Salmonella typhimurium if proteome pre-fractionation steps and gas-phase fractionation were included.     

The effect of kinetin on the photosynthetic apparatus of Sinapis alba

The influence of kinetin during the development of primary leaves of Sinapis alba was investigated. Kinetin treatment (6 ppm) induced an increase of dry weight, of soluble reducing sugars, soluble protein, chlorophylls, carotenoids and cytochrome f; a higher ratio of chlorophyll a to chlorophyll b, higher rates of CO₂ fixation per fresh weight and higher activity of nitrite reductase, were also found. These effects are comparable with strong and blue light adaptations. On the other hand, the Hill activity with ferricyanide as the electron acceptor, the rates of CO₂ fixation per chlorophyll, the ratios of chlorophyll to cytochrome f and of protein to chlorophyll did not change. Therefore we assume that the kinetin induced and the light induced adaptations are brought about by different causal reaction chains.

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Zusammenfassung Es wurde die Wirkung von Kinetin auf die Entwicklung von Primarblattern von Senfpflanzen untersucht. Die Behandlung mit Kinetin (6 ppm) bewirkte eine Erhöhung des Trochengewichtes, der Gehalte an löslichen, reduzierend wirkenden Zuckern, an löslichem Protein, Chlorophyllen, Karotinoiden und Cytochrom f, sowie eine Erhöhung des Quotienten von Chlorophyll a zu Chlorophyll b, eine verstärkten Einbau von CO₂ pro Frischgewicht und eine Erhöhung der Nitritreduktase-Aktivität. Diese Auswirkungen sind den durch Starklicht und Blaulicht hervorgerufenen Anpassungsreaktionen vergleichbar. Andererseits zeigten die Hill-Reaktion (gemessen als Reduktion von Ferricyanid), die CO₂ Fixierung pro Chlorophyll, der Quotient von Chlorophyll zu Cytochrom f und der Quotient von Protein zu Chlorophyll keire Veränderungen. Dies weist darauf hin, daß die durch Kinetin und durch Licht hervorgerufenen Anpassungsreaktionen durch verschiedene Kausalketten bedingt werden.

     

Anti-inflammatory properties of intestinal Bifidobacterium strains isolated from healthy infants

Certain Bifidobacterium strains have been shown to inhibit inflammatory responses in intestinal epithelial cells. However, the precise mechanisms of these effects, including the chemical nature of the active compounds, remain to be elucidated. Here partial characterization of the anti-inflammatory properties of Bifidobacterium strains isolated from feces of healthy infants is reported. It was found that conditioned media (CM) of all strains studied are capable of attenuating tumor necrosis factor-α (TNF-α) and lipopolysaccharide- (LPS) induced inflammatory responses in the HT-29 cell line. In contrast, neither killed bifidobacterial cells, nor cell-free extracts showed such activities. Further investigations resulted in attribution of this activity to heat-stable, non-lipophilic compound(s) resistant to protease and nuclease treatments and of molecular weight less than 3 kDa. The anti-inflammatory effects were dose- and time-dependent and associated with inhibition of IκB phosphorylation and nuclear factor-κ light chain enhancer of activated B cells (NF-κB)-dependent promoter activation. The combined treatments of cells with CMs and either LPS or TNF-α, but not with CMs alone, resulted in upregulation of transforming growth factor-β1, IκBζ, and p21(CIP) mRNAs. Our data suggest certain species-specificities of the anti-inflammatory properties of bifidobacteria. This observation should prompt additional validation studies using larger set of strains and employing the tools of comparative genomics.     

基于光投影数字化传感系统的飞蝗表面三维结构

本文基于结构光投影数字化传感系统构建东亚飞蝗Locusta migratoria manilensis表面的三维结构。首先得到其外形尺寸和肢体各部分拓扑结构,然后从被调制的条纹结构光中恢复出物体表面高度变化的相位信息,再通过相位和高度的对应关系重建出物体表面三维特征。此结果将为农药研制和施用设计提供科学依据,对促进蝗虫的防治以及虚拟昆虫研究起到积极作用。     

不同光质对烟草叶片组织结构及Rubisco羧化酶活性和rbc、rca基因表达的影响

通过对烟草植株覆盖白、红、黄、蓝、紫色滤膜获得不同光质,研究了烟草叶片在7～70d的生长发育期内,不同光质处理对烟叶组织结构特征、核酮糖1,5-二磷酸羧化酶/加氧酶（Rubisco）羧化酶活性、Rubisco基因（rbc）表达及其活化酶（Rca）基因（rca）表达的影响。结果表明,与黄膜处理下生长的烟叶相比,红、蓝、紫膜处理下生长的烟叶有较高的叶片厚度、栅栏组织厚度、海绵组织厚度、栅栏细胞密度和较小的组织空隙率。此外,红、蓝、紫膜处理的叶片有较高的Rubis-co羧化酶活性和净光合速率及较强的rbc和rca基因表达。实验结果表明不同光质对烟草叶片的组织结构特征有显著影响,光质可能通过影响Rubisco羧化酶活性进而影响叶片光合效率,而光质、叶片组织结构和光合效率之间存在某种程度的相互联系。     

Co-adaptation of seed dormancy and flowering time in the arable weed Capsella bursa-pastoris (shepherd's purse)

Background and Aims

The duration of the plant life cycle is an important attribute that determines fitness and coexistence of weeds in arable fields. It depends on the timing of two key life-history traits: time from seed dispersal to germination and time from germination to flowering. These traits are components of the time to reproduction. Dormancy results in reduced and delayed germination, thus increasing time to reproduction. Genotypes in the arable seedbank predominantly have short time to flowering. Synergy between reduced seed dormancy and reduced flowering time would create stronger contrasts between genotypes, offering greater adaptation in-field. Therefore, we studied differences in seed dormancy between in-field flowering time genotypes of shepherd''s purse.

Methods

Genotypes with early, intermediate or late flowering time were grown in a glasshouse to provide seed stock for germination tests. Secondary dormancy was assessed by comparing germination before and after dark-incubation. Dormancy was characterized separately for seed myxospermy heteromorphs, observed in each genotype. Seed carbon and nitrogen content and seed mass were determined as indicators of seed filling and resource partitioning associated with dormancy.

Key Results

Although no differences were observed in primary dormancy, secondary dormancy was weaker among the seeds of early-flowering genotypes. On average, myxospermous seeds showed stronger secondary dormancy than non-myxospermous seeds in all genotypes. Seed filling was similar between the genotypes, but nitrogen partitioning was higher in early-flowering genotypes and in non-myxospermous seeds.

Conclusions

In shepherd''s purse, early flowering and reduced seed dormancy coincide and appear to be linked. The seed heteromorphism contributes to variation in dormancy. Three functional groups of seed dormancy were identified, varying in dormancy depth and nitrate response. One of these groups (FG-III) was distinct for early-flowering genotypes. The weaker secondary dormancy of early-flowering genotypes confers a selective advantage in arable fields.