Spindle disturbances in human-hamster hybrid (AL) cells induced by mobile communication frequency range signals

The production of spindle disturbances in FC2 cells, a human-hamster hybrid (A(L)) cell line, by non-ionizing radiation was studied using an electromagnetic field with a field strength of 90 V/m at a frequency of 835 MHz. Due to the given experimental conditions slide flask cultures were exposed at room temperature in a microTEM (transversal electromagnetic field) cell, which allows optimal experimental conditions for small samples of biological material. Numerical calculations suggest that specific absorption rates of up to 60 mW/kg are reached for maximum field exposure. All exposure field parameters--either measured or calculable--are precisely defined and, for the first time, traceable to the standards of the SI system of physical units. Compared with co-incident negative controls, the results of two independently performed experiments suggest that exposure periods of time from 0.5 to 2 h with an electric field strength of 90 V/m are spindle acting agents as predominately indicated by the appearance of spindle disturbances at the ana- and telophase stages (especially lagging and non-disjunction of single chromosomes) of cell divisions. The spindle disturbances do not change the fraction of mitotic cells with increasing exposure time up to 2 h. Due to the applied experimental conditions an influence of temperature as a confounder parameter for spindle disturbances can be excluded.     

中国梨2个自交不亲和新等位基因(S等位基因)的分子鉴定

自交不亲和是显花植物的一种重要生殖生理现象,为探明中国梨的自交不亲和特性,对‘锦香’(Pyrus bretschneideri cv. Jinxiang)和‘鹅酥’(Pyrus bretschneideri cv. Esu)2个中国梨品种进行了基因组PCR特异扩增、S基因序列分析及田间杂交授粉试验。结果确定它们各含1个新S-RNA酶基因,分别命名为S37-和S38-RNase,GenBank序列号为DQ839238和DQ839239。生物信息学分析结果表明,S37-和S38-RNA酶的推导氨基酸序列与S1-至S36-RNA酶36个梨S基因具有相同的、高度保守的C1和C2区,但其高变区与S1-至S36-RNA酶差异较大,其中与S15的差异最小,只有3个氨基酸不同。在推导的氨基酸水平上,S37与S38有96%的序列相似性,但两者与S15的相似性更高,皆为98%,与S32的相似性最低,都只有63%;S37和S38的内含子较大,分别为786bp和723bp,与S15的777bp大小接近。最后,经分析验证确定‘锦香’和‘鹅酥’的S基因型分别为S34S37和S15S38。     

Ezetimibe markedly attenuates hepatic cholesterol accumulation and improves liver function in the lysosomal acid lipase-deficient mouse,a model for cholesteryl ester storage disease

Lysosomal acid lipase (LAL) plays a critical role in the intracellular handling of lipids by hydrolyzing cholesteryl esters (CE) and triacylglycerols (TAG) contained in newly internalized lipoproteins. In humans, mutations in the LAL gene result in cholesteryl ester storage disease (CESD), or in Wolman disease (WD) when the mutations cause complete loss of LAL activity. A rat model for WD and a mouse model for CESD have been described. In these studies we used LAL-deficient mice to investigate how modulating the amount of intestinally-derived cholesterol reaching the liver might impact its mass, cholesterol content, and function in this model. The main experiment tested if ezetimibe, a potent cholesterol absorption inhibitor, had any effect on CE accumulation in mice lacking LAL. In male Lal^−/− mice given ezetimibe in their diet (20 mg/day/kg bw) for 4 weeks starting at 21 days of age, both liver mass and hepatic cholesterol concentration (mg/g) were reduced to the extent that whole-liver cholesterol content (mg/organ) in the treated mice (74.3 ± 3.4) was only 56% of that in those not given ezetimibe (133.5 ± 6.7). There was also a marked improvement in plasma alanine aminotransferase (ALT) activity. Thus, minimizing cholesterol absorption has a favorable impact on the liver in CESD.     

Behavioral plasticity and virus propagation: the FIV-cat population example

A new theoretical approach is proposed to investigate the effect of intra-individual variability in behavior on the spread of directly transmitted diseases within host populations. The classical hawk-dove game is used to describe interactions between individuals on a fast time scale (the day). Individuals may exhibit both tactics according to their own experience, to environment conditions, and to the opponent. They are not able to recognize the epidemiological state of their opponents. This fast-time part of the model is coupled to a classical compartmental epidemic model describing the demography of the population and the transmission of the disease from an infected individual to a healthy one on a slow time scale (the year). The model is applied to the case of feline immunodeficiency virus (FIV)-domestic cat population system. Our model gives rise to different predictions according to values of cost and gain due to fights: extinction of the epidemic, FIV endemicity at low, intermediate and high prevalence. These predictions are in good agreement with results from domestic cat populations living in different environmental conditions.     

Structure,gene expression,and evolution of primate copper chaperone for superoxide dismutase

Copper chaperone for superoxide dismutase (CCS) is essential for transporting copper ion to Cu,Zn-superoxide dismutase (Cu,Zn-SOD). We cloned cDNAs for six primate species' CCSs. The total number of amino acid residues of primate CCSs is 274. Similarities between primates were over 96%. Important residues for the CCS function were well conserved. A phylogenetic tree of CCSs and Cu,Zn-SODs from various organisms showed that these two proteins were derived from a common ancestor, diverging very early on during eukaryote evolution. The high frequency of nonsynonymous substitutions was found in the lineage to Old World monkeys and apes. Expression of the CCS gene in various tissues of Japanese monkey was found to be high in the liver and adrenal gland, followed by the kidney and small intestine. Such expressional pattern was similar with that of Cu,Zn-SOD gene (Fukuhara et al., 2002).     

Functional Properties of Slowly Adapting Mechanoreceptors in Cat Footpad Skin

The functional properties of slowly adapting (SA) afferent fibers innervating cat footpad skin were examined. Measurements were taken of receptive field area; spontaneous activity (< 1 impulse/sec); the slope of the stimulus-response curve for steady indentations up to 2 mm in amplitude; variability of the interimpulse intervals, as measured by the coefficient of variation of time interval histograms; decay of the response to steady indentation; and sensitivity to sinusoidal vibration (most sensitive at 5-10 Hz). Where comparable tests were performed on glabrous and hairy skin SA fibers, the functional properties of those in glabrous skin more closely resembled SAI fibers than SAII fibers. Additional results from glabrous skin SA fibers suggest that it is distortion of the nerve endings rather than steady indentation or compression that leads to a brisk response. On the measures described above, there appeared to be only one functional class of SA fiber innervating the cat footpad skin.     

Quantitative evaluation of respiration induced metabolic oscillations in erythrocytes

The changes in the partial pressures of oxygen and carbon dioxide (P_O2 and P_CO2) during blood circulation alter erythrocyte metabolism, hereby causing flux changes between oxygenated and deoxygenated blood. In the study we have modeled this effect by extending the comprehensive kinetic model by Mulquiney and Kuchel [P.J. Mulquiney, and P.W. Kuchel. Model of 2,3-bisphosphoglycerate metabolism in the human erythrocyte based on detailed enzyme kinetic equations: equations and parameter refinement, Biochem. J. 1999, 342, 581–596.] with a kinetic model of hemoglobin oxy-/deoxygenation transition based on an oxygen dissociation model developed by Dash and Bassingthwaighte [R. Dash, and J. Bassingthwaighte. Blood HbO₂ and HbCO₂ dissociation curves at varied O₂, CO₂, pH, 2,3-DPG and temperature levels, Ann. Biomed. Eng., 2004, 32(12), 1676–1693.]. The system has been studied during transitions from the arterial to the venous phases by simply forcing P_O2 and P_CO2 to follow the physiological values of venous and arterial blood. The investigations show that the system passively follows a limit cycle driven by the forced oscillations of P_O2 and is thus inadequately described solely by steady state consideration. The metabolic system exhibits a broad distribution of time scales. Relaxations of modes with hemoglobin and Mg²⁺ binding reactions are very fast, while modes involving glycolytic, membrane transport and 2,3-BPG shunt reactions are much slower. Incomplete slow mode relaxations during the 60 s period of the forced transitions cause significant overshoots of important fluxes and metabolite concentrations – notably ATP, 2,3-BPG, and Mg²⁺. The overshoot phenomenon arises in consequence of a periodical forcing and is likely to be widespread in nature – warranting a special consideration for relevant systems.     

Application of solid-phase Ellman''s reagent for preparation of disulfide-paired isomers of α-conotoxin SI

Disulfide-paired regioisomers of -conotoxin SIcan be accessed by orthogonal schemes using thecombination of S-9H-xanthen-9-yl (S-Xan) and S-acetamidomethyl (S-Acm)groups for cysteine protection. Following solid-phaseassemblies of the linear precursors, the peptides werecleaved from the solid support concurrent with removalof S-Xan protecting groups. The first disulfidebridges were formed in solution, using either thetraditional DMSO method or a recently introducedapproach featuring a solid-phase Ellman's reagent. The second disulfide bridges were oxidized by threedifferent methods: reactions mediated by thalliumtrifluoroacetate, iodine, or a sulfoxide/silylmixture. In general, yields depended primarily onwhich regioisomer was the target, rather than thespecific chemistry used for either disulfide-formingstep. However, the selectivities towards the desiredregio-isomers were reproducibly better using thesolid-phase Ellman's reagent, by comparison to theDMSO method. In the most favorable cases, completeselectivity was achieved, while even in cases wherethe net results using DMSO gave considerablescrambling, the corresponding experiments with thesolid-phase Ellman's reagent were more selective. Possible reasons why choice of oxidation method forthe first step affects the selectivity at the secondstep are discussed.     

Application of solid-phase Ellman's reagent for preparation of disulfide-paired isomers of α-conotoxin SI

Summary Disulfide-paired regioisomers of α-conotoxin SI can be accessed by orthogonal schemes using the combination ofS-9H-xanthen-9-yl (S-Xan) andS-acetamidomethyl (S-Acm) groups for cysteine protection. Following solidphase assemblies of the linear precursors, the peptides were cleaved from the solid support concurrent with removal ofS-Xan protecting groups. The first disulfide bridges were formed in solution, using either the traditional DMSO method or a recently introduced approach featuring a solid-phase Ellman's reagent. The second disulfide bridges were oxidized by three different methods: reactions mediated by thallium trifluoroacetate, iodine, or a sulfoxide/silyl mixture. In general, yields depended primarily on which regioisomer was the target, rather than the specific chemistry used for either disulfide-forming step. However, the selectivities towards the desired regioisomers were reproducibly better using the solid-phase Ellman's reagent, by comparison to the DMSO method. In the most favorable cases, complete selectivity was achieved, while even in cases where the net results using DMSO gave considerable scrambling, the corresponding experiments with the solid-phase Ellman's reagent were more selective. Possible reasons why choice of oxidation method for the first step affects the selectivity at the second step are discussed.