Methionine anchoring applied to the solid-phase synthesis of lysine-containing 'head-to-tail' cyclic peptides

Summary Lysine-containing ‘head-to-tail’ cyclic peptides can be prepared via a side-chain anchoring solid-phase synthesis strategy. A handle is prepared using a methionine residue, theC ^α-carboxyl of which forms an, amide with theN ^ε-amine of lysine. Subsequently, the linear peptide sequence is assembled, appropriate deblocking steps are carried out, and on-resin head-to-tail cyclization follows. Optionally, acid-labile protecting groups may be removed while the peptide remains resin-bound. The final cleavage step uses CNBr, and releases the free or protected cyclic peptide into solution. Taken in part from the Ph.D. Thesis of J. C. Kappel, University of Minnesota, November 2003. Portions of this work were reported in preliminary form at the Eighteenth American Peptide Symposium, Boston, MA, U.S.A., 19–23 July 2003, and at the Eighth International Symposium on Solid Phase Synthesis and Combinatorial Chemical Libraries, London, England, U.K., 2–5 September 2003.     

Remodeling of nuclear architecture during the cell cycle in Drosophila embryos

Little is known about what determines the nuclear matrix or how its reorganization is regulated during mitosis. In this study we report on a monoclonal antibody, mAb2A, which identifies a novel nuclear structure in Drosophila embryos which forms a diffuse meshwork at interphase but which undergoes a striking reorganization into a spindle-like structure during pro- and metaphase. Double labelings with α-tubulin and mAb2A antibodies demonstrate that the microtubules of the mitotic apparatus co-localize with this mAb2A labeled structure during metaphase, suggesting it may serve a role in microtubule spindle assembly and/or function during nuclear division. That the mAb2A-labeled nuclear structure is essential for cell division and/or maintenance of nuclear integrity was directly demonstrated by microinjection of mAb2A into early syncytial embryos which resulted in a disintegration of nuclear morphology and perturbation of mitosis. © 1996 Wiley-Liss, Inc.     

Estimating sampling error of evolutionary statistics based on genetic covariance matrices using maximum likelihood

We explore the estimation of uncertainty in evolutionary parameters using a recently devised approach for resampling entire additive genetic variance–covariance matrices ( G ). Large‐sample theory shows that maximum‐likelihood estimates (including restricted maximum likelihood, REML) asymptotically have a multivariate normal distribution, with covariance matrix derived from the inverse of the information matrix, and mean equal to the estimated G . This suggests that sampling estimates of G from this distribution can be used to assess the variability of estimates of G , and of functions of G . We refer to this as the REML‐MVN method. This has been implemented in the mixed‐model program WOMBAT. Estimates of sampling variances from REML‐MVN were compared to those from the parametric bootstrap and from a Bayesian Markov chain Monte Carlo (MCMC) approach (implemented in the R package MCMCglmm). We apply each approach to evolvability statistics previously estimated for a large, 20‐dimensional data set for Drosophila wings. REML‐MVN and MCMC sampling variances are close to those estimated with the parametric bootstrap. Both slightly underestimate the error in the best‐estimated aspects of the G matrix. REML analysis supports the previous conclusion that the G matrix for this population is full rank. REML‐MVN is computationally very efficient, making it an attractive alternative to both data resampling and MCMC approaches to assessing confidence in parameters of evolutionary interest.     

核基质结合区对转爬山虎芪合酶基因烟草中产生白藜芦醇的作用

采用核基质结合区(MARs)来提高转芪合酶基因(STS)烟草(Nicotianatabacum L.)中白藜芦醇产物的含量.MARs是细胞中能与核基质特异紧密结合的DNA片段,体外结合实验表明克隆自酵母的MARs序列能特异地与烟草核基质结合.芪合酶是白藜芦醇生物合成中的关键酶,用RT-PCR方法从川鄂爬山虎(Parthenocissus henryana(Hemsl.)Diels et Gilg)中克隆了与葡萄芪合酶基因有较高同源性的芪合酶编码区,将其置于CaMV35SΩ强启动子下,分别构建两侧带有MARs及不含MARs序列的表达载体,通过农杆菌介导转化烟草.Northern blot及HPLC等分析表明STS基因已整合至烟草染色体中并正常转录,且表达的外源芪合酶在烟草中可催化其底物合成白藜芦醇产物.与对照相比,MARs的存在使转芪合酶基因烟草中白藜芦醇的含量平均提高了约一倍.MARs在转芪合酶基因植物中的应用也为获得抗病性更强、白藜芦醇含量更高、更保健的转基因果蔬的研究奠定了基础.     

To identify impacts in variable systems using anomalous changes: a salt marsh example

A method is proposed to identify impacts of habitat modification in cases where it is difficult to site experimental and control samples. This problem occurs especially in heterogeneous systems, but may pose difficulties in any field experimental situation. The method is relevant to the situation where treated (modified) and untreated sites are spread over a range of habitat types. Types of change are identified and compared to treatments. If a specific change type is associated with a particular treatment then it is likely that the change is causally related to the treatment. There are five stages in the analysis. First, the classes or states of the sample sites, over a period of time, are identified (by numerical classification). Second, for each sample site, the sequence of states is listed. Third, transition matrices are made for each sample site to show the changes which have occurred. Fourth, the transition matrices are classified, to identify types of change. Finally, we use the Chisquared test to indicate whether the treated and untreated sites are associated with particular types of change. As an example, we refer to habitat modification to manage salt-marsh mosquitoes and we evaluate impacts on the environment mainly through changes to the vegetation. We consider that the method has potential to identify changes in heterogeneous systems even though little change was identified in the particular salt marsh studied.     

The hidden face of food phenolic composition

Plant polyphenols are extremely diverse, due to the occurrence of several basic structures, numerous substitutions and, for some groups, of polymers (tannins). Plant polyphenol composition depends on the plant species and organ, with some molecules specific of particular plant families while others are ubiquitous. The polyphenol content is classically assessed by global analysis methods, which lack specificity and accuracy. These methods have been replaced with high performance liquid chromatography (HPLC), that enables accurate determination of individual molecules, provided they can be unambiguously identified and calibration curves can be established. However, HPLC analysis is restricted to simple compounds and difficult to apply in the case of complex extracts. Further difficulties encountered in the case of polymers include irreversible adsorption on the stationary phases. Proanthocyanidin analysis by HPLC after acid-catalysed depolymerisation in the presence of a nucleophile permits to overcome these problems and shows that proanthocyanidins predominate in the polyphenol composition of most plants. Large varietal differences in tannin quantitative and qualitative composition were observed for all plant species studied. Moreover, analysis is usually performed after extraction, which may lead to significant underestimation of the polyphenol content, since a large proportion is not extracted by usual solvents. This may be due to covalent binding to other plant constituents and to non-covalent adsorption on plant solids. Such matrix effect also influences the taste perception of polyphenols and their fate in the digestive tract, from in-mouth interactions with salivary proteins to their metabolism by colon microflora, with potential influence on bioavailability.     

Nuclear structure as a source of cancer specific biomarkers

There are few biomarkers that have been developed which have proven clinical utility for the detection and prognosis of cancer. Cancer is diagnosed today, in large part, by examining cells under the microscope and determining the shape and texture of the nucleus. The molecular underpinnings of this hallmark of cancer are the components of the nuclear matrix. Utilizing proteomics focused on this subset of proteins, biomarkers have been identified that are specific for cancer types including prostate, colon and bladder cancer. These cancer biomarkers now serve as the basis of assays which can specifically identify individuals with cancer by sampling their blood and/or urine. In addition, these may serve as potential therapeutic targeting or imaging approaches.     

Reconsidering locomotor habits and life style of the Balearic insular giant rodent Hypnomys Bate, 1918 from the allometry of the limb long bones

The genus Hypnomys Bate, 1918 includes some endemic Neogene chronospecies from Mallorca and Menorca, evolved in insularity conditions from the Lower Pliocene to the human arrival at the archipelago. The study of the allometric proportions (functional length and sagittal diameter) of the limbs’ long bones of Hypnomys eliomyoides Agusti, 1980 (Lower Pleistocene of Menorca), Hypnomys sp. (Pleistocene of Menorca), Hypnomys onicensis (Reumer, 1994) (Upper Pliocene-Lower Pleistocene of Mallorca) and Hypnomys morpheus Bate, 1918 (Upper Pleistocene of Mallorca) only indicates small differences with the garden dormouse (Eliomys quercinus Linnaeus, 1776) and other mainland rodents and insectivores. The study about the locomotion type by means of Similarity Matrix of Bou indicates that 1) Hypnomys spp. presents the greater similarities with the arboreal, jumping and gliding species; 2) the jumping and digging life style of Hypnomys spp. are more developed (25%) than in E. quercinus; 3) the walking locomotion of Hypnomys spp. is only a 7% greater than in E. quercinus; and 4) the degree of similarity with the gliding locomotion of Hypnomys sp. is greater (17%) than in E. quercinus. Some of this data indicates that Hypnomys spp. was better adapted to the arboreal life than E. quercinus, due to the relationship between the jumping and gliding capacities with the arboreal locomotion. The greater similarity of Hypnomys spp. with the walking locomotion does not necessarily involve more terrestrial habits. Our interpretation contrasts with earlier conclusions of Hypnomys life style (Hypnomys cf. onicensis and Hypnomys morpheus) where the terrestrial locomotion was overestimated in front of the arboreal locomotion. The locomotion type of Hypnomys would be related with the presence of important forest masses on the Balearic Islands during the Neogene, a different degree of environmental stress from that of the mainland ecosystems and a probable expansion of the ecological niche occupied by these species.