Human haptoglobin type and G-streptococci: a safe and unexpected relation]

By testing 167 human sera a surprising dimorphism could be found. All sera of the haptoglobin type hp 1-1 belong to the group with an anti-G streptococci titer in the range of 1:0 to 1:10 whereas sera of the types hp 2-1 and hp 2-2 do have titers in the range between 1:200 and 1:3200 or more. All tests are carried out in the double blind technique.     

Tandem GA residues on opposite sides of the loop in molecular beacon-like DNA hairpins compact the loop and increase hairpin stability

The free solution electrophoretic mobilities and thermal stabilities of hairpins formed by two complementary 26-nucleotide oligomers have been measured by capillary electrophoresis. The oligomers are predicted to form molecular beacon-like hairpins with 5 bp stems and 16 nucleotides in the loop. One hairpin, called hairpin2 (hp2), migrates with a relatively fast free solution mobility and exhibits melting temperatures that are reasonably well predicted by the popular structure-prediction program Mfold. Its complement, called hairpin1 (hp1), migrates with a slower free solution mobility and forms a stable hairpin only in solutions containing ≥200 mM Na(+). The melting temperatures observed for hp1 are ~18 °C lower than those observed for hp2 and ~20 °C lower than those predicted by Mfold. The greater thermal stability of hp2 is due to the presence of tandem GA residues on opposite sides of the loop. If the corresponding TC residues in the hp1 loop are replaced by tandem GA residues, the melting temperatures of the modified hairpin are close to those observed for hp2. Eliminating the tandem GA residues in the hp2 loop significantly decreases the thermal stability of hp2. If the loops are replaced by a loop of 16 thymine residues, the free solution mobilities and thermal stabilities of the T-loop hairpin are equal to those observed for hp1. Hence, the loop of hp1 appears to be relatively unstructured, with few base-base stacking interactions. Interactions between tandem GA residues on opposite sides of the hp2 loop appear to compact the loop and increase hairpin stability.     

Three New Cutinases from the Yeast Arxula adeninivorans That Are Suitable for Biotechnological Applications

The genes ACUT1, ACUT2, and ACUT3, encoding cutinases, were selected from the genomic DNA of Arxula adeninivorans LS3. The alignment of the amino acid sequences of these cutinases with those of other cutinases or cutinase-like enzymes from different fungi showed that they all had a catalytic S-D-H triad with a conserved G-Y-S-Q-G domain. All three genes were overexpressed in A. adeninivorans using the strong constitutive TEF1 promoter. Recombinant 6× His (6h)-tagged cutinase 1 protein (p) from A. adeninivorans LS3 (Acut1-6hp), Acut2-6hp, and Acut3-6hp were produced and purified by immobilized-metal ion affinity chromatography and biochemically characterized using p-nitrophenyl butyrate as the substrate for standard activity tests. All three enzymes from A. adeninivorans were active from pH 4.5 to 6.5 and from 20 to 30°C. They were shown to be unstable under optimal reaction conditions but could be stabilized using organic solvents, such as polyethylene glycol 200 (PEG 200), isopropanol, ethanol, or acetone. PEG 200 (50%, vol/vol) was found to be the best stabilizing agent for all of the cutinases, and acetone greatly increased the half-life and enzyme activity (up to 300% for Acut3-6hp). The substrate spectra for Acut1-6hp, Acut2-6hp, and Acut3-6hp were quite similar, with the highest activity being for short-chain fatty acid esters of p-nitrophenol and glycerol. Additionally, they were found to have polycaprolactone degradation activity and cutinolytic activity against cutin from apple peel. The activity was compared with that of the 6× His-tagged cutinase from Fusarium solani f. sp. pisi (FsCut-6hp), also expressed in A. adeninivorans, as a positive control. A fed-batch cultivation of the best Acut2-6hp-producing strain, A. adeninivorans G1212/YRC102-ACUT2-6H, was performed and showed that very high activities of 1,064 U ml⁻¹ could be achieved even with a nonoptimized cultivation procedure.     

Antioxidant Properties of New Chalcogenides Against Lipid Peroxidation in Rat Brain

Ebselen (2-phenyl- 1,2-benzisoselenazole-3 (2H)-one) is a seleno-organic compound with antioxidant properties, and anti-inflammatory actions. Recently, ebselen improved the outcome of acute ischemic stroke in humans. In the present study, the potential antioxidant capacity of organochalcogenide compounds diphenyl diselenide (PhSe)₂, diphenyl ditelluride (PhTe)₂, diphenyl disulfide (PhS)₂, p-Cl-diphenyl diselenide (pCl-PhSe)₂, bis-[S-4-isopropyl 2-phenyl oxazoline] diselenide (AA-Se)₂, bis-[S-4-isopropyl 2-phenyl oxazoline] ditelluride (AA-Te)₂ and bis-[S-4-isopropyl 2-phenyl oxazoline] disulfide (AA-S)₂ was compared with that of ebselen (a classical antioxidant). Spontaneous and quinolinic acid (QA)- (2 mM) and sodium nitroprusside (SNP)- (5 M)-induced thiobarbituric reactive species (TBARS) production by rat brain homogenates was determined colorimetrically. TBARS formation was reduced by ebselen, (PhSe)₂, (PhTe)₂, (AA-Se)₂, (AA-S)₂ and (pCl- PhSe)₂ to basal rates. The concentrations of these compounds needed to inhibit TBARS formation by 50% (lC₅₀) are 1.71 M, 3.73 M, 1.63 M, 9.85 M, > 33.3 M, 23.2 M and 4.83 M, respectively for QA. For TBARS production induced by SNP the lC₅₀ was 2.02 M, 12.5 M, 2.80 M, > 33.3 M, 24.5 M and 7.55 M, respectively. The compounds (AA-Te)₂ and (PhS)₂ have no antioxidant activity and pro-oxidant activity, respectively. These results suggest that (AA-Se)₂ and (AA-S)₂ can be considered as potential pharmaceutical antioxidant agents.     

Enhanced transpiration rate in the high pigment 1 tomato mutant and its physiological significance

Tomato high pigment (hp) mutants represent an interesting horticultural resource due to their enhanced accumulation of carotenoids, flavonoids and vitamin C. Since hp mutants are known for their exaggerated light responses, the molecules accumulated are likely to be antioxidants, recruited to deal with light and others stresses. Further phenotypes displayed by hp mutations are reduced growth and an apparent disturbance in water loss. Here, we examined the impact of the hp1 mutation and its near isogenic line cv Micro-Tom (MT) on stomatal conductance (gs), transpiration (E), CO(2) assimilation (A) and water use efficiency (WUE). Detached hp1 leaves lost water more rapidly than control leaves, but this behaviour was reversed by exogenous abscisic acid (ABA), indicating the ability of hp1 to respond to this hormone. Although attached hp1 leaves had enhanced gs, E and A compared to control leaves, genotypic differences were lost when water was withheld. Both instantaneous leaf-level WUE and long-term whole plant WUE did not differ between hp1 and MT. Our results indicate a link between exaggerated light response and water loss in hp1, which has important implications for the use of this mutant in both basic and horticultural research.     

High-affinity binding of fungal β-glucan elicitors to cell membranes of species of the plant family Fabaceae

Microsomal preparations of six species of the plant family Fabaceae were screened for high-affinity binding of branched (1 3), (1 6)--glucans. Oligoglucosides of this type are specific elicitors of phytoalexin accumulation in soybean (Glycine max L.), a member of this family. The species studied were alfalfa (Medicago sativa L.), broadbean (Vicia faba L.), chickpea (Cicer arietinum L.), french bean (Phaseolus vulgaris L.), pea (Pisum sativum L.), and white lupin (Lupinus albus L.). A ¹²⁵I-labeled 4-(2-aminophenyl)ethylamine conjugate of a (1 3), (1 6)--glucan fraction with an average degree of polymerization (DP) of 18, obtained from mycelial walls of Phytophthora sojae, was used as radioligand for initial screening. The structural complexity of this fraction allowed the identification of binding sites with affinities for isomeric structures other than the (1 3), (1 6) hepta--glucoside for which soybean binding sites display highest affinity. Radioligand competition experiments against unlabeled fungal -glucan resulted in the identification of high-affinity binding in alfalfa, bean, lupin, and pea. Half-maximal competition concentrations (IC₅₀) for fungal -glucan in these species were between 5 and 30 nM. Pseudoheterologous radioligand competition by unlabeled hepta--glucoside showed that for alfalfa, lupin and pea the IC₅₀ values for this structure (4 to 16 nM) were similar to those of soybean (7.7 nM). Bean microsomes, however, displayed an IC₅₀ significantly higher than soybean (68 nM) suggesting that the structural motif recognized by its binding sites is not identical to that of soybean or the other three species. Radioligand saturation assays with alfalfa, lupin and pea microsomes using an ₁₂₅I-labeled aminophenylethylamine hepta--glucoside conjugate gave dissociation constants (K_d) of 5.3, 3.7, and 1.8 nM, respectively. The affinity of these sites for hepta- glucoside was in the same range as that of soybean (K_d 1–3 nM), whereas the affinity of the binding sites of bean for the same ligand was significantly lower (K_d = 33 nM). Good correlation was found between the presence of high-affinity binding and the accumulation of isoflavonoid phytoalexins in roots of alfalfa, bean, chickpea and pea seedlings after exposure to fungal -glucan. Lupin displayed a strong wound-induced accumulation of prenylated isoflavones which was independent of the presence of -glucan, making it impossible to determine phytoalexin induction in response to elicitor. No specific binding or phytoalexin accumulation in response to glucans was observed in broadbean. This is the first report on the existence of possibly homologous elicitor-binding sites within a plant taxonomic family and may provide preliminary evidence for putative evolutionary relationships in pathogen perception mechanisms in plants.Abbreviations DP degree of polymerization - EC₅₀ concentration of elicitor necessary to obtain a half-maximal biological response - HG synthetic (1 3), (1 6)-hepta--glucoside phytoalexin elicitor - HG-APEA 1-[4-(2-aminophenyl)ethylamino-1-hexaglucosyl]deoxyglucitol - IC₅₀ ligand concentration necessary to obtain half-maximal displacement of radioligand in competition binding assays - K_d dissociation constant - OS branched (1 3), (1 6)--glucan obtained by hydrolysis of mycelial walls of Phytophthora sojae - OS-APEA 1-[4-(2-armnophenyl)ethylamino-1-oligoglucosyl]deoxyglucitol conjugate of OS This work was supported by the Comision Interministerial de Ciencia y Tecnologia grant BI091-0366 (E.G.C.), the Volkswagen-Stiftung (E.G.C. and J.E.), the Deutsche Forschungsgemeinschaft, SFB-369 (J.E.), the Bundesministerium fiir Bildung, Wissenschaft, Forschung und Technologie (J.E.), Fonds der Chemischen Industrie (J.E.) and the EU Human Capital and Mobility Program (J.E. and E.G.C.).     

Yield and post-harvest quality of tomato hybrids heterozygous at the loci alcobaça, old gold-crimson or high pigment

The effects of the fruit ripening mutant gene alcoba?a (alc) and color development mutants, old gold-crimson (ogc) and high pigment (hp), on yield and post-harvest quality of tomato fruits were investigated. Five tomato hybrids were obtained by crossing near isogenic lines with Flora-Dade background [Flora-Dade (alc+/alc+ ogc+/ogc+ hp+/hp+), TOM-559 (alc/alc ogc+/ogc+ hp+/hp+), TOM-591 (alc/alc ogc/ogc hp+/hp+), TOM-593 (alc/alc ogc+/ogc+ hp/hp), and TOM-589 (alc/alc ogc/ogc hp/hp)] with the pollen parent line Mospomorist (alc+/alc+ ogc+/ogc+ hp+/hp+). Hybrid fruit was harvested at the breaker stage and stored on shelves at 15oC and 60% relative humidity for 16 days, and then evaluated for firmness, development of red color, and carotenoid contents. The different genotypic combinations at the loci alc, ogc and hp had no effect on fruit yield. The alc+/alc hybrid genotype significantly increased fruit firmness and significantly delayed the development of red color in maturing fruit. Simultaneous usage of ogc+/ogc and hp+/hp promoted an increase in the red color and lycopene content of alc+/alc hybrids, but did not have any additional effect on fruit firmness.     

Altered proteome profiles in maternal plasma in pregnancies with fetal growth restriction

Fetal growth restriction (FGR) affects 3–5% of pregnancies and is associated with increased perinatal morbidity and mortality. Currently, there is no reliable biochemical test to differentiate a pathological FGR from a nonpathological one. The objective of this study was to screen whole maternal plasma to identify differentially expressed relatively abundant proteins associated with FGR. We analyzed maternal plasma from FGR (n=28) and healthy (n=22) pregnancies using two-dimensional gel electrophoresis (2D-GE) followed by software image analysis. Three spots with molecular weight (M_r) 18 kDa corresponding to haptoglobin (hp) α2, as identified by LC-MS/MS and immunoblotting, showed differential expression patterns in FGR. The distribution of hp α2 variants in maternal plasma samples showed the hp α2 variant 1 was low in 72% of FGR, medium in 16%, whereas high in 12%. In comparison, hp α2 variant 1 was high in (41%) of controls, medium in 41%, and low in 18% of cases. Based on the software image analysis, the mean spot volume for hp α2 variant 1 was 0.12 (SD=0.18) for FGR compared to 0.26 (SD=0.19) for control (p=0.006). Given that hp turnover is indicative of its maturation process and is traceable in plasma by its dominant/suppressed variants, we propose that hp α2 is an important potential target for evaluation of its clinical and pathophysiological role and as a diagnostic biomarker in FGR.     

Suitability of the shaking flask for oxygen supply to microbiological cultures

Due of its simplicity the shaking flask is used in serial studies, e.g. in the screening for secondary metabolites or in the optimization of fermentation processes. Experimental investigations in these small bioreactors are often the first step in developing a large-scale fermentation process.Movement of the flask should produce sufficient mixing, supply of oxygen, and removal of carbon dioxide. In the case of fluids with low or moderate viscosity, gas transport is the most important aspect. This publication summarizes data necessary to calculate the gas transport. These data are derived from the consideration of the gas diffusions through the cotton plug as well as from the substance transport between the gas and liquid phases. As a result suitable fermentation conditions can be selected. Finally, the performance limits of the shaking flask are illustrated using the example of the oxygen supply in a Streptomyces tendae fermentation.List of Symbols A _s Cross section of plug - A Surface area of liquid in flask - a A/V _F specific phase interface area - c Concentration - c ^* Saturation concentration - c Plug diffusion term - D Widest diameter of flask - Diffusion coefficients in multicomponent gas mix tures - Diffusion coefficients in binary gas mixtures - Diffusion coefficient of oxygen in the liquid - d Diameter of neck of flask - e Eccentricity - G Volume-based mass flow - G _m Maximum volume-based mass flow - g Acceleration due to gravity - h Height coordinate - ¯H Mean height of plug - Hy p _i/c ^*, Henry constant - K Consistency index - k D _xy/D _xz, Ratio of diffusion coefficients in binary gas mixtures - k _M Monod constant - k _L a Mass transport coefficient: gas/liquid - M Molecular weight - m Flow exponent - n Speed of shaking - p Pressure - p _i Partial pressure of gas component i - q Area-based flow of volume - R , respiration ratio - Sc , Schmidt number - T Absolute temperature - V Flask volume - V _F Volume of liquid in flask - w Velocity of the Stefan flow - x, y, z Ratios of the partial pressures of the gases O₂, CO₂, N₂ - Rate of shear - Dynamic viscosity of the liquid - Kinematic viscosity of the liquid - Density of the liquid - _x, Density of O₂ gas - Surface tension Indices 0 State in gas volume of shaking flask - 1 State in outside air - G Gas volume - x, y, z O₂, CO₂, N₂     

Morphology and physiology of a hair plate sensory organ located on the antenna of the rock lobster Palinurus vulgaris

At the level of the J1 joint of each antenna of the rock lobster Palinurus vulgaris a hair plate sensory organ (hp) similar to those described in insects has been observed. The hp is located on the internal side of the S2 segment of the antenna, close to the soft articulating membrane of the J1 joint. It is formed by a triangular surface of cuticle about 3mm2 in area, covered with numerous hairs of different lengths (Figs. 1 and 2). Details of the hp were studied by scanning electron microscopy (Fig. 2). Physiological stimulation of the hp hairs occurs during medial movement of the J1 joint. Under this condition the soft articulating membrane rolls over the hairs and bends them progressively back onto the cuticle. Flexion of all the hairs corresponds to a medial movement of the J1 through 40 degrees. During this type of movement, the number of successively flexed hairs increases linearly (Fig. 3). Electrophysiological recordings of the hp sensory nerve correlated with selective mechanical stimulation of individual hairs demonstrated that each hair is innervated by a single sensory fiber. This sensory neurone responds phasically when the hair is flexed back onto the cuticle (as during an S2 medial movement) and when it returns to its resting position (as during an S2 lateral movement). Most of the sensory neurones are sensitive to the movement velocity of the hairs (Figs. 4 and 5). When the hair is maintained flexed its sensory neurone discharges tonically (Fig. 4). Electrical stimulation of the hp sensory nerve induced reflex actions in the external and internal rotator muscles of the segment S1. These effects were found to selectively activate the tonic motor command of these muscles (Fig. 6).     

glmm.hp: an R package for computing individual effect of predictors in generalized linear mixed models

glmm.hp:一个计算广义线性混合模型中单个预测变量效应的R包 广义线性混合效应模型(GLMMs)是当代生态学研究中广泛应用的数据分析模型。然而,确定GLMMs中共线性的预测变量(固定效应)对响应变量的相对重要性是个挑战。基于适用于多元分析的‘平均共享方差’的算法,我们开发一个新的R包glmm.hp来分解GLMMs中由固定效应解释的边际(marginal)R²。我们论述了该软件包的工作原理并通过模拟数据集演示了该软件包的使用。glmm.hp 包的输出结果为每个预测变量将获得一个独自的(individual)边际R²,且它们的总和刚好等于模型总的边际R²。总之,我们相信glmm.hp包将有助于解释GLMMs 的输出结果。     

Estimation of specific growth rate from agitation speed in DO-stat culture

Summary A simple and effective method to estimate the specific growth rate estimation has been developed based on the observation of time changes in the agitation speed in dissolved oxygen(DO)-stat cultures of Brevibacterium ketoglutamicum. The estimation was compared with that using carbon dioxide evolution rate (CER). Estimated values of specific growth rates by both methods agreed well with the data directly calculated from cell concentration change although the use of agitation speed gave a slightly better result than CER.Nomenclature CER Carbon dioxide evolution rate (mmol/sec) - OUR Oxygen uptake rate (mmol/sec) - OTR Oxygen transfer rate (mmol/sec) - RPM Agitation speed (rev./min) - C* Saturated dissolved oxygen concentration (mmol/L) - Dissolved oxygen concentration (mmol/L) - k Time index - k _L a' Mass transfer coefficient (sec^-1) - Y _X/O2 Cellular yield based on oxygen consumed (g-cell/mmol O₂) - Specific growth rate (hr^-1) - Constant - t Fermentation time - t Sampling time for RPM and CER measurements