The effects of exogenous proline and proline analogues on in vitro shoot organogenesis in Arabidopsis

In vitro shoot organogenesis from Arabidopsis hypocotyl explants was stimulated by 1 mMproline, and to a lesser extent by 5 mM proline, butinhibited by inclusion of 10 mM proline in thehormonally-supplemented regeneration medium. Theability of low concentrations of the prolineanalogues, azetidine-2-carboxylate and thioproline toovercome the stimulatory effect of 1 mM proline, anda slight increase in the stimulative effects of 1 mMproline by D-proline, are consistent with an importantrole for the interconversions of proline and itsprecursors in regulating cell division anddifferentiation.     

Effects of resistance training on physical function in older disabled women with coronary heart disease.

We studied whether disabled older women with coronary heart disease can perform resistance training at an intensity sufficient to improve measured and self-reported physical function [n = 30, 70.6 +/- 4.5 (SD) yr]. Compared with the controls, the resistance-training group showed significant improvements in overall measured physical function score using the Continuous-Scale Physical Functional Performance Test (+24 vs. +3%). The Continuous-Scale Physical Functional Performance Test measures physical function for 15 practical activities, such as carrying groceries or climbing stairs. Resistance training led to improved measures for domains of upper body strength (+18 vs. +6%), lower body strength (+23 vs. +6%), endurance (+26 vs. +1%), balance and coordination (+29 vs. -2%), and 6-min walk (+15 vs. +7%). Women involved in the flexibility-control group showed essentially no improvement for physical function measures. No changes were observed for body composition, aerobic capacity, or self-reported physical function in either group. In conclusion, disabled older women with coronary heart disease who participate in strength training are able to train at an intensity sufficient to result in improvements in multiple domains of measured physical functional performance, despite no change in lean body mass.     

Non-thermal exposure to radiofrequency energy from digital wireless phones does not affect ornithine decarboxylase activity in L929 cells

L929 murine fibroblast cells were exposed to radiofrequency (RF) radiation from a time division multiple access wireless phone operating at 835 MHz frequency to determine the effect of RF-radiation energy emitted by wireless phones on ornithine decarboxylase (ODC) activity in cultured cells. Exposure was for 8 h to an average specific absorption rate (SAR) from <1 W/kg up to 15 W/kg. After exposure, cells were harvested and ODC activity was measured. No statistically significant difference in ODC activity was found between RF-radiation-exposed and sham-exposed cells at non-thermal specific absorption rates. At SARs which resulted in measurable heating of the medium, a dose-dependent decrease in enzymatic activity was observed and was shown to be consistent with a comparable decrease caused by non-RF-radiation heating. Thus we observed only the well-known enzyme inhibition due to heating, rather than the previously reported enhancement attributed to RF-radiation exposure.     

Evidence for dynamic alteration in histone gene clusters of Caenorhabditis elegans: a topoisomerase II connection?

Chromatin integrity is maintained throughout the cell cycle through repair mechanisms and intrinsically by the ordered packaging of DNA in association with histone proteins; however, aberrant rearrangements within and between chromosomes do occur. The role of the nuclear matrix protein topoisomerase II (TopoII) in generating chromosome breakpoints has been a focus of recent investigations. TopoII preferentially binds in vitro to scaffold-associated regions (SARs) and is involved in many DNA processing activities that require chromosome untangling. SARs, biochemically defined DNA elements rich in A + T, have been proposed to serve as structural boundaries for chromatin loops and to delineate functional domains. In our investigation of gene compartmentalization in a eukaryotic genome, SAR-associated nucleotide motifs from Drosophila were mapped in the regions of three histone gene clusters in an in silico analysis of the genome of Caenorhabditis elegans. Sites with similarity to the 15 bp consensus for TopoII cleavage were found predominantly in A + T enriched intergenic regions. Reiteration of sites matching the TopoII core consensus led to the identification of a novel core histone gene on chromosome IV and provided evidence for duplication and inversion in each of the three histone gene clusters. Breakpoint analysis of DNA flanking reiterated regions revealed potential sites for TopoII cleavage and a base composition phenomenon suggestive of a trigger for inversion events.     

Quantitation of phosphotyrosine signals in human prostate cell adhesion sites

A simple method is described for the quantitation of phosphotyrosine signaling in human prostate cell cultures. The phosphotyrosine signals are observed by standard immunohistochemistry techniques, and the resulting digital images are analyzed using the Scion image software program. The signals within the cell adhesion sites are quantitated using the density slice and particle analysis features of the software. The immunohistochemistry results are compared with detection of phosphotyrosine signals using a standard Western blotting procedure with whole cell lysates. The resulting data is converted into graphs using the Sigma Plot Program. This method is illustrated using damage-induced signaling within cell adhesion sites after a low dose of ionizing radiation.     

Nonglutamate pore residues in ion selection and conduction in voltage-gated Ca(2+) channels

High-affinity, intrapore binding of Ca(2+) over competing ions is the essential feature in the ion selectivity mechanism of voltage-gated Ca(2+) channels. At the same time, several million Ca(2+) ions can travel each second through the pore of a single open Ca(2+) channel. How such high Ca(2+) flux is achieved in the face of tight Ca(2+) binding is a current area of inquiry, particularly from a structural point of view. The ion selectivity locus comprises four glutamate residues within the channel's pore. These glutamates make unequal contributions to Ca(2+) binding, underscoring a role for neighboring residues in pore function. By comparing two Ca(2+) channels (the L-type alpha(1C), and the non-L-type alpha(1A)) that differ in their pore properties but only differ at a single amino acid position near the selectivity locus, we have identified the amino-terminal neighbor of the glutamate residue in motif III as a determinant of pore function. This position is more important in the function of alpha(1C) channels than in alpha(1A) channels. For a systematic series of mutations at this pore position in alpha(1C), both unitary Ba(2+) conductance and Cd(2+) block of Ba(2+) current varied with residue volume. Pore mutations designed to make alpha(1C) more like alpha(1A) and vice versa revealed that relative selectivity for Ba(2+) over K(+) depended almost solely on pore sequence and not channel type. Analysis of thermodynamic mutant cycles indicates that the motif III neighbor normally interacts in a cooperative fashion with the locus, molding the functional behavior of the pore.     

拼接信号序列单碱基变异提高马传染性贫血病毒mRNA拼接效率

分别以马传染性贫血(马传贫)驴强毒(D—A EIAV)RNA和马传贫驴白细胞弱毒疫苗(DLA EIAV)RNA为模板,利用RT—PCR的方法,克隆到马传贫强、弱毒株基因组外显子2及其下游的核苷酸序列。然后将报告基因CAT插入到EIAV内含子2env阅读框架中,构成CAT拼接报告系统。同时在强毒株重组表达质粒的基础上,将其外显子-3上游拼接受体位点的核苷酸序列CAG突变为弱毒株相应位置的核苷酸序列TAG,得到强毒单核苷酸突变株重组表达质粒。用构建的3个重组表达质粒DNA转染驴血白细胞,ELISA检测转染细胞CAT浓度。结果表明：EIAV强毒株重组表达质粒中CAT蛋白表达量最高,EIAV强毒株重组表达质粒次之,EIAV强毒突变株重组表达质粒最低。由于CAT基因被插入于各重组质粒中的EIAV内含子-2里,EIAV外显子-2、3之间的拼接可导致该基因的删除,因而其拼接效率低于EIAVmRNA外显子-2、3之间的拼接效率。实验数据表明,EIAV SA2拼接信号序列单碱基变异提高了SD2-SA2拼接效率;D—AEIAV SA2-SD2拼接效率比DLA EIAV相应位点拼接效率高。