Ultrastructural localization of adenosine triphosphatase activity in HeLa cells at various stages of the cell cycle

Summary HeLa cells in a monolayer culture were synchronized to S, G₂ and mitotic phases by use of excess (2.5 mM) deoxythymidine double-block technique. The localizations of Ca⁺⁺-activated adenosine triphosphatase (ATPase) at different phases of the cell cycle were studied using light and electron-microscopic histochemical techniques, and microphotometric comparisons of the densities of reaction products. Enzyme reaction product was always localized in the endoplasmic reticulum, nuclear membrane, mitochondria and Golgi apparatus, but there were qualitative and quantitative differences related to the phases of the cell cycle. In S phase the activity was mainly concentrated in a perinuclear area of the cytoplasm whereas in G₂ and mitosis the activity was scattered throughout the cell. The total activity per cell was maximal in G₂, was less in S phase and least in mitosis. Activity in the mitochondria and endoplasmic reticulum was distinctly less in mitosis than in other phases of the cell cycle. The mitochondrial ATPase differed from the ATPase at other sites in ion dependence and sensitivity to oligomycin. The results suggest that there may be several distinct ATPases in proliferating cells.     

The effect of a daytime 60-min nap opportunity on postural control in highly active individuals

Although napping is commonly used as a strategy to improve numerous physical and cognitive performances, the efficacy of this strategy for improving postural balance has not yet been elucidated. Thus, the aim of this study was to conduct a comprehensive examination of the effect of a 60 min nap opportunity (N60) on different components of postural control. Ten highly active individuals (age = 27 ± 3.5 y, height = 1.75 ± 0.52 m, weight = 66.02 ± 8.63 kg) performed, in a randomized order, two afternoon test sessions following no nap (NN) and N60. Postural balance was assessed using the sensory organisation test (SOT), the unilateral stance test (UST), and the limits of Stability Test performed on NeuroCom^® Smart Balance Master. The subjective rating of sleepiness before and after the nap conditions was also assessed. Compared to NN, N60 improved the composite balance score (p < 0.05, ES = 0.75, Δ = 5.3%) and the average and maximum percentage balance in the most challenging postural conditions of the SOT (p < 0.05 for SOT-4 and 5 and p < 0.0005 for SOT-6; ES range between 0.58 and 1.1). This enhanced postural balance in N60 was accompanied with improved visual (p < 0.05; ES = 0.93; Δ = 8.9%) and vestibular (p < 0.05; ES = 0.81; Δ = 10.5%) ratios and a reduced level of sleepiness perception (p < 0.001, ES = 0.87). However, no significant differences were found in any of the UST and LOS components’ scores (p > 0.05). Overall, a 60 min post lunch nap opportunity may be viable for improving static balance, although further work, involving larger samples and more complex motor activities, is warranted.     

Clostridium tunisiense sp. nov., a new proteolytic, sulfur-reducing bacterium isolated from an olive mill wastewater contaminated by phosphogypse

A new sporulated fermentative bacterium designated strain E1(T) (T=type strain), was isolated from an anaerobic mud of an olive mill wastewater basin contaminated by phosphogypse produced by a Tunisian factory. Strain E1(T) was a motile Gram-positive slightly curved rod with spherical terminal spore swelling the cell. It grew between 18 degrees C and 43 degrees C with an optimum at 37 degrees C and pH 7.8 (range 5.5-8.7), without NaCl (range 0-3%). Strain E1(T) was a chemoorganotrophic anaerobic bacterium fermenting only proteins and amino acids. Yeast extract was required for growth. Elemental sulfur was used as terminal electron acceptor. The G+C content of the DNA was 32.6 mol%. The closest phylogenetical relatives of strain E1(T) were Clostridium thiosulfatireducens and C. subterminale (97.3% similarity for partial rRNA gene sequences). DNA-DNA hybridization values between strain E1(T) and both species were 17% and 20.8%, respectively. On the basis of differences in genotypic and phenotypic characteristics, strain E1(T) (DSM 15206(T), CIP 107666(T)) is proposed as the type strain of a new species, C. tunisiense sp. nov. GenBank accession number for the 16S rRNA gene sequence of strain E1(T) is AY187622.     

突尼斯两种麦瘿蚊的遗传变异和相关性分析

小麦黑森瘿蚊Mayetiola destructor Say和大麦茎干瘿蚊M. hordei Kieffer是在突尼斯每年都可导致谷物重大损失的两个植食性姊妹种。通常认为为害小麦的瘿蚊是小麦黑森瘿蚊,但是不同谷类物种（小麦或大麦）与麦瘿蚊的这两个种（ destructor 或 hordei ）之间寄主关系并不很严格。提出有效的害虫管理方案首先要求对瘿蚊基因型进行精确分析。本研究应用随机扩增多态DNA（RAPD）技术,结合交配分析和线粒体DNA分型技术,对位于突尼斯北部的一个为害小麦的麦瘿蚊种群的遗传变异程度和分类关联性进行了评估。基于RAPD结果的系统发育分析表明,所研究的种群具有较大的遗传变异范围,这可能源于被分析的小麦样品有被两种瘿蚊共同侵害的复杂背景。虽然交配分析表明有少数不能成功产卵(2/14),但是基于细胞色素b基因限制性酶切分析显示全部样品的线粒体分型均属M. destructor。本文结果进一步证明以RAPD可变性作为分类推断依据不可靠,还为突尼斯M. destructor和M. hordei属于异域分布的观点提供了补充证据。     

A new species ofConiochaetidium from soil

A new species ofConiochaetidium isolated from soil of Iraq is described. The speciesC. nuciforme differs fromC. savoryi in the shape and size of the ascospores. A key to differentiate the accepted species of the genus is reported.     

Synthesis, characterisation and solution behaviour of fluoroalkyl phosphoryl complexes of tin tetrachloride

The synthesis, characterisation and solution behaviour of a series of octahedral complexes SnCl₄·2L (L = R₂NP(O)(OCH₂CF₃)₂; R = Me (1); Et (2) or L = P(O)(OCH₂R_f)₃; R_f = CF₃ (3); C₂F₅ (4)) are described. Complexes 1-4 were prepared from SnCl₄ and 2 equiv. of the ligand, L, in anhydrous CH₂Cl₂ solution. The adducts have been characterised by multinuclear (¹H, ³¹P and ¹¹⁹Sn) NMR, IR spectroscopy and elemental analysis. In dichloromethane solution, the NMR data showed the presence of a mixture of cis and trans isomers for 1 and 2 and only the cis isomer for 3 and 4. The difference could be interpreted in terms of the electronic effects of the substituents on the phosphorus atom of the ligand. In addition, the solution structure of the complexes studied by variable temperature ³¹P-{¹H} and ¹H NMR in the presence of excess ligand indicated that the ligand exchange on the cis isomer dominates the chemistry. The metal-ligand exchange barriers were estimated to be 13.38 and 11.39 kcal/mol for 1 and 3, respectively. The results are discussed and compared with those previously reported for the related hexamethylphosphoramide adduct, SnCl₄·2HMPA.     

<Emphasis Type="Italic">Desulfovibrio legallis</Emphasis> sp. nov.: A Moderately Halophilic,Sulfate-Reducing Bacterium Isolated from a Wastewater Digestor in Tunisia

A new moderately halophilic sulfate-reducing bacterium (strain H₁^T) was enriched and isolated from a wastewater digestor in Tunisia. Cells were curved, motile rods (2–3 x 0.5 μm). Strain H₁^T grew at temperatures between 22 and 43°C (optimum 35°C), and at pH between 5.0 and 9.2 (optimum 7.3–7.5). Strain H₁^T required salt for growth (1–45 g of NaCl/l), with an optimum at 20–30 g/l. Sulfate, sulfite, thiosulfate, and elemental sulfur were used as terminal electron acceptors but not nitrate and nitrite. Strain H₁^T utilized lactate, pyruvate, succinate, fumarate, ethanol, and hydrogen (in the presence of acetate and CO₂) as electron donors in the presence of sulfate as electron acceptor. The main end-products from lactate oxidation were acetate with H₂ and CO₂. The G + C content of the genomic DNA was 55%. The predominant fatty acids of strain H₁^T were C_15:0 iso (38.8%), C_16:0 (19%), and C_14:0 iso 3OH (12.2%), and menaquinone MK-6 was the major respiratory quinone. Phylogenetic analysis of the small-subunit (SSU) ribosomal RNA (rRNA) gene sequence indicated that strain H₁^T was affiliated to the genus Desulfovibrio. On the basis of SSU rRNA gene sequence comparisons and physiological characteristics, strain H₁^T is proposed to be assigned to a novel species of sulfate reducers of the genus Desulfovibrio, Desulfovibrio legallis sp. nov. (= DSM 19129^T = CCUG 54389^T).     

The multi-mode polarization modulation spectrometer: part 1: simultaneous detection of absorption, turbidity, and optical activity

Circular dichroism (CD) is an important spectroscopic technique for monitoring chirality and biological macromolecule conformation. However, during a CD measurement, absorbance, light scattering/turbidity, and fluorescence can also be detected. The simultaneous measurement of these different spectral features for a single sample is the basis of a multi-mode optical spectrometer. This allows time-efficient gathering of complementary information and provides a scheme to ensure that CD measurements are reliable. Aspects of circular polarization differential light scattering, pH, and temperature variation of a protein (antibody) solution are described. A procedure to help ensure that CD measurements are reliable is described.     

Wistar Rats Resistant to the Hypertensive Effects of Ouabain Exhibit Enhanced Cardiac Vagal Activity and Elevated Plasma Levels of Calcitonin Gene-Related Peptide

Ouabain is a cardiac glycoside produced in the adrenal glands and hypothalamus. It affects the function of all cells by binding to Na⁺/K⁺-ATPase. Several lines of evidence suggest that endogenous ouabain could be involved in the pathogenesis of essential (particularly, salt-sensitive) hypertension. However, information regarding the postulated hypertensive effect of the long-term administration of low-dose exogenous ouabain is inconsistent. This study was designed to help settle this controversy through the use of telemetric monitoring of arterial blood pressure and to elucidate the ouabain-induced alterations that could either promote or prevent hypertension. Ouabain (63 and 324 µg/kg/day) was administered subcutaneously to male Wistar rats. Radiotelemetry was used to monitor blood pressure, heart rate and measures of cardiovascular variability and baroreflex sensitivity. The continuous administration of ouabain for 3 months did not elevate arterial blood pressure. The low-frequency power of systolic pressure variability, urinary excretion of catecholamines, and cardiovascular response to restraint stress and a high-salt diet as well as the responsiveness to α1-adrenergic stimulation were all unaltered by ouabain administration, suggesting that the activity of the sympathetic nervous system was not increased. However, surrogate indices of cardiac vagal nerve activity based on heart rate variability were elevated. Molecular remodeling in mesenteric arteries that could support the development of hypertension (increased expression of the genes for the Na⁺/Ca²⁺ exchanger and Na⁺/K⁺-ATPase α2 isoform) was not evident. Instead, the plasma level of vasodilatory calcitonin gene-related peptide (CGRP) significantly rose from 55 (11, SD) in the control group to 89 (20, SD) pg/ml in the ouabain-treated rats (P_Tukey''s = 18.10⁻⁵). These data show that long-term administration of exogenous ouabain does not necessarily cause hypertension in rodents. The augmented parasympathetic activity and elevated plasma level of CGRP could be linked to the missing hypertensive effect of ouabain administration.