X-ray microanalysis of elements in frozen-hydrated sections of an electrogenic K+ transport system: The posterior midgut of tobacco hornworm (Manduca sexta) in vivo andin vitro

Summary The lepidopteran midgut is a model for the oxygendependent, electrogenic K⁺ transport found in both alimentary and sensory tissues of many economically important insects. Structural and biochemical evidence places the K⁺ pump on the portasome-studded apical plasma membrane which borders the extracellular goblet cavity. However, electrochemical evidence implies that the goblet cell K⁺ concentration is less than 50mm. We used electron probe X-ray microanalysis of frozenhydrated cryosections to measure the concentration of Na, Mg, P, S, Cl, K, Ca and H₂O in several subcellular sites in the larval midgut ofManduca sexta under several experimental regimes. Na is undetectable at any site. K is at least 100mm in the cytoplasm of all cells. Typicalin vivo values (mm) for K were: blood, 25; goblet and columnar cytoplasm, 120; goblet cavity, 190; and gut lumen, 180. The high K concentration in the apically located goblet cavity declined by 100mm under anoxia. Both cavity and gut fluid are Cl deficient, but fixed negative charges may be present in the cavity. We conclude that the K⁺ pump is sited on the goblet cell apical membrane and that K⁺ follows a nonmixing pathway via only part of the goblet cell cytoplasm. The cavity appears to be electrically isolated in alimentary tissues, as it is in sensory sensilla, thereby allowing a PD exceeding 180 mV (lumen positive) to develop across the apical plasma membrane. This PD appears to couple K⁺ pump energy to nutrient absorption and pH regulation.     

Evaluation of support matrices for immobilization of anaerobic consortia for efficient carbon cycling in waste regeneration

Efficient metabolism of fatty acids during anaerobic waste digestion requires development of consortia that include "fatty acid consuming H(2) producing bacteria" and methanogenic bacteria. The objective of this research was to optimize methanogenesis from fatty acids by evaluating a variety of support matrices for use in maintaining efficient syntrophic-methanogenic consortia. Tested matrices included clays (montmorillonite and bentonite), glass beads (106 and 425-600mum), microcarriers (cytopore, cytodex, cytoline, and cultispher; conventionally employed for cultivation of mammalian cell lines), BioSep beads (powdered activated carbon), and membranes (hydrophilic; nylon, polysulfone, and hydrophobic; teflon, polypropylene). Data obtained from headspace methane (CH(4)) analyses as an indicator of anaerobic carbon cycling efficiency indicated that material surface properties were important in maintenance and functioning of the anaerobic consortia. Cytoline yielded significantly higher CH(4) than other matrices as early as in the first week of incubation. 16S rRNA gene sequence analysis from crushed cytoline matrix showed the presence of Syntrophomonas spp. (butyrate oxidizing syntrophs) and Syntrophobacter spp. (propionate oxidizing syntrophs), with Methanosaeta spp. (acetate utilizing methanogen), and Methanospirillum spp. (hydrogen utilizing methanogen) cells. It is likely that the more hydrophobic surfaces provided a suitable surface for adherence of cells of syntrophic-methanogenic consortia. Cytoline also appeared to protect entrapped consortia from air, resulting in rapid methanogenesis after aerial exposure. Our study suggests that support matrices can be used in anaerobic digestors, pre-seeded with immobilized or entrapped consortia on support matrices, and may be of value as inoculant-adsorbents to rapidly initiate or recover proper system functioning following perturbation.     

Pairwise versus presence–absence approaches for analysing biodiversity patterns

It has been proposed that the study of co‐occurrence of species, which is traditionally performed using full presence–absence matrices of sets of many species, could benefit from simply testing for random co‐occurrence between pairs of species, and that use of a full presence–absence matrix is tantamount to regarding it as having some real ecological identity. Here I argue that although there are valid questions that can be answered using a pairwise approach, there are many others that naturally require the analysis of entire sets of species in a joint way, as provided for through the use of full presence–absence matrices. Moreover, there are theoretical and mathematical advantages to the use of presence–absence matrices, a few of which are briefly discussed in this short note.     

Tissue engineered nerve constructs:where do we stand?

Driven by enormous clinical need, interest in peripheral nerve regeneration has become a prime focus of research and area of growth within the field of tissue engineering. While using autologous donor nerves for bridging peripheral defects remains today's gold standard, it remains associated with high donor site morbidity and lack of full recovery. This dictates research towards the development of biomimetic constructs as alternatives. Based on current concepts, this review summarizes various approaches including different extracellular matrices, scaffolds, and growth factors that have been shown to promote migration and proliferation of Schwann cells. Since neither of these concepts in isolation is enough, although each is gaining increased interest to promote nerve regeneration, various combinations will need to be identified to strike a harmonious balance. Additional factors that must be incorporated into tissue engineered nerve constructs are also unknown and warrant further research efforts. It seems that future directions may allow us to determine the "missing link".     

MODELING BRAIN EVOLUTION FROM BEHAVIOR: A PERMUTATIONAL REGRESSION APPROACH

This paper has two complementary purposes: first, to present a method to perform multiple regression on distance matrices, with permutation testing appropriate for path-length matrices representing evolutionary trees, and then, to apply this method to study the joint evolution of brain, behavior and other characteristics in marsupials. To understand the computation method, consider that the dependent matrix is unfolded as a vector y; similarly, consider X to be a table containing the independent matrices, also unfolded as vectors. A multiple regression is computed to express y as a function of X. The parameters of this regression (R² and partial regression coefficients) are tested by permutations, as follows. When the dependent matrix variable y represents a simple distance or similarity matrix, permutations are performed in the same manner as the Mantel permutational test. When it is an ultrametric matrix representing a dendrogram, we use the double-permutation method (Lapointe and Legendre 1990, 1991). When it is a path-length matrix representing an additive tree (cladogram), we use the triple-permutation method (Lapointe and Legendre 1992). The independent matrix variables in X are kept fixed with respect to one another during the permutations. Selection of predictors can be accomplished by forward selection, backward elimination, or a stepwise procedure. A phylogenetic tree, derived from marsupial brain morphology data (28 species), is compared to trees depicting the evolution of diet, sociability, locomotion, and habitat in these animals, as well as their taxonomy and geographical relationships. A model is derived in which brain evolution can be predicted from taxonomy, diet, sociability and locomotion (R² = 0.75). A new tree, derived from the “predicted” data, shows a lot of similarity to the brain evolution tree. The meaning of the taxonomy, diet, sociability, and locomotion predictors are discussed and conclusions are drawn about the evolution of brain and behavior in marsupials.     

Bis(pyrazolyl) palladium(II), platinum(II) and gold(III) complexes: Syntheses, molecular structures and substitution reactions with l-cysteine

A series of pyrazolyl palladium(II), platinum(II) and gold(III) complexes, [PdCl₂(3,5-R₂bpza)] {R = H (1), R = Me (2), bpza = bis-pyrazolyl acetic acid}, [PtCl₂(3,5-R₂bpza)] {R = H (3a), R = Me (4)}, [AuCl₂(3,5-R₂bpza)]Cl {R = H (5a), R = Me (6a)} and [PdCl₂(3,5-R₂bpzate)] {R = Me (7)} have been synthesised and structurally characterised. Single crystal X-ray crystallography showed that the pyrazolyl ligands exhibit N^N-coordination with the metals. Anticancer activities of six complexes 1-6a were investigated against CHO cells and were found to have low activities. Substitution reactions of selected complexes 1, 2, 3a and 5a with l-cysteine show that the low anticancer activities compounds and that the rate of substitution with sulfur-containing compounds is not the cause of the low anticancer activities.     

单链构象多态性毛细管电泳分析研究进展

基因突变的检测在临床疾病诊断中起十分重要的作用.单链构象多态性(Single Strand Conform ationPolym orphism,SSCP)分析是检测突变最流行的方法之一.SSCP分析与毛细管电泳(Capillary Electrophoresis,CE)相结合的技术更是具有灵敏度高、花费低、简单、快速的优点.目前,这项技术已应用于人类原癌基因、抑癌基因以及其它致病基因的突变检测.主要综述了各种参数对CES-SCP分析的影响以及CES-SCP分析技术将来的发展方向.     

Adaptation to slow environmental change, with apparent anticipation of selection

We investigate a genetic model of a large population of sexual organisms in a changing environment. The organisms are subject to stabilising selection on a quantitative trait, with environmental change causing the fitness optimum to move. When the fitness optimum moves slowly, adaptation to the changing environment occurs by means of reasonably well-separated substitutions at the loci controlling the trait. In this way, the trait generally tracks the moving optimum, but in such a case, the population may exhibit periods of time where the mean trait value overshoots the moving optimal trait value, thereby exhibiting an apparent anticipation of selection. The mechanism underlying this phenomenon is determined from consideration of a simpler model that correctly captures the observed dynamical behaviour. We note that very slow rates of changes of traits are seen in the fossil record and the present work may be relevant to this topic.     

Pairwise null model analyses of temporal patterns of bird assemblages contradict the assumptions of competition theory

Compensatory dynamics assume inverse patterns of population dynamics of species with similar ecological resource requirements (temporal segregation). The objective of this study was to test this hypothesis on temporal samples (10–57 years) of 19 breeding bird assemblages of various habitats. We used presence/absence null model (SIM2) in combination with the C-score and Sørensen indices. The C-score index estimates the average number of checkerboards for two species, while the Sørensen index measures the qualitative similarity of co-occurrence between two species in a time series. We used pairwise null model analysis to select significant species pairs based on three selection criteria: the standard confidence interval criterion, conservative empirical Bayes mean based criterion and confidence limit based criterion. Altogether, 21 402 species pairs were analysed. The SIM2 algorithm detected from 157 to 7 segregated pairs depending on the selection criterion. The number of significant negative pairs with possible biological significance (foraging guild membership, predator–prey interactions) was far lower and represented approximately 0.0–0.3% (4–65) of pairs in a matrix. Indeed, the number of detected negative associations depended on the selection criterion. Moreover, the number of segregated pairs was negatively related to the area of the census plots and fill of the species matrix. Our results underline the minor importance of interspecific competitive interactions in temporal patterns of bird assemblages. Instead, we suggest that stochastic factors, climate or heterospecific social information may lead to more or less synchronous dynamics of bird pairs.     

P-related sequences inDrosophila bifasciata: A molecular clue to the understanding of P-element evolution in the genusDrosophila

Summary Two P-elements (bif1 and bif2) were isolated from a genomic library ofDrosophila bifasciata. Both elements are internally deleted and have lost the coding capacity for a functional transposase. One of the elements (bif2) contains an insert consisting of a repetitive sequence. The terminal inverted repeats and the segments necessary for passive mobility are well conserved. Element bif2 has retained rudiments of the coding sequence of exon 0 and exon 3, but the reading frame is destroyed by insertions and deletions. The comparison of theD. bifasciata P-elements with P-elements ofDrosophila melanogaster andDrosophila nebulosa reveals that the two latter sequences are more similar to each other than either of them is to theD. bifasciata elements. This finding contradicts the phylogenetic relationship of the species and can be taken as an indirect but unequivocal evidence for recent horizontal gene transfer from a relative ofD. nebulosa to the gene pool ofD. melanogaster. The P-elements ofD. bifasciata are phylogenetically ancient and have evolved independently for about 50 million years. A higher substitution rate at the third codon position as well as a predominance of conservative replacements at the amino acid level indicates that the P-elements ofD. bifasciata have been under selective constraint over a long period and that immobilization has occurred only recently.