Development of a gas chromatography mass spectrometry assay for the two eosinophil chemotactic factors of anaphylaxis

Gas chromatography mass spectrometry has been used for over a decade for the determination of the amino acid sequences of fragment peptides derived from larger parent molecules. The majority of these fragments have from four to seven residues and several different methods of derivatization have been devised. Few reports have been published in which similar techniques have been used for the quantification of such peptides, but there is a growing list of small peptides which have been shown to have biological activity in their own right. This report is concerned with the development of a gas chromatographic mass spectrometric assay for the two eosinophil chemotactic peptides, Val-Gly-Ser-Glu and Ala-Gly-Ser-Glu, which appear to have a role to play in the course of the inflammatory process in skin disorders.     

Means and extremes: building variability into community‐level climate change experiments

Experimental studies assessing climatic effects on ecological communities have typically applied static warming treatments. Although these studies have been informative, they have usually failed to incorporate either current or predicted future, patterns of variability. Future climates are likely to include extreme events which have greater impacts on ecological systems than changes in means alone. Here, we review the studies which have used experiments to assess impacts of temperature on marine, freshwater and terrestrial communities, and classify them into a set of ‘generations’ based on how they incorporate variability. The majority of studies have failed to incorporate extreme events. In terrestrial ecosystems in particular, experimental treatments have reduced temperature variability, when most climate models predict increased variability. Marine studies have tended to not concentrate on changes in variability, likely in part because the thermal mass of oceans will moderate variation. In freshwaters, climate change experiments have a much shorter history than in the other ecosystems, and have tended to take a relatively simple approach. We propose a new ‘generation’ of climate change experiments using down‐scaled climate models which incorporate predicted changes in climatic variability, and describe a process for generating data which can be applied as experimental climate change treatments.     

Prevacuolar compartment morphology in vps mutants of Saccharomyces cerevisiae

Over 60 genes have been identified that affect protein sorting to the lysosome-like vacuole in Saccharomyces cerevisiae. Cells with mutations in these vacuolar protein sorting (vps) genes fall into seven general classes based upon their vacuolar morphology. Class A mutants have a morphologically wild type vacuole, while Class B mutants have a fragmented vacuole. There is no discernable vacuolar structure in Class C mutants. Class D mutants have a slightly enlarged vacuole, but Class E mutants have a normal looking vacuole with an enlarged prevacuolar compartment (PVC), which is analogous to the mammalian late endosome. Class F mutants have a wild type appearing vacuole as well as fragmented vacuolar structures. vps mutants have also been found with a tubulo-vesicular vacuole structure. vps mutant morphology is pertinent, as mutants of the same class may work together and/or have a block in the same general step in the vacuolar protein sorting pathway. We probed PVC morphology and location microscopically in live cells of several null vps mutants using a GFP fusion protein of Nhx1p, an Na(+)/H(+) exchanger normally localized to the PVC. We show that cell strains deleted for VPS proteins that have been previously shown to work together, regardless of VPS Class, have the same PVC morphology. Cell strains lacking VPS genes that have not been implicated in the same pathway show different PVC morphologies, even if the mutant strains are in the same VPS Class. These new studies indicate that PVC morphology is another tier of classification that may more accurately identify proteins that function together in vacuolar protein sorting than the original vps mutation classes.     

Core-coat conductor of lipid bilayer and micromachined silicon

We have etched a groove into a (110) plane of silicon and have covered it with a bilayer of glycerol monooleate. We have varied the depth of the groove, the concentration of salt in the electrolyte and the density of gramicidin in the membrane. We have clamped one end of the groove at a constant voltage with respect to the bath keeping the other end sealed or electrically open with respect to the bath. We have measured (i) the voltage at the center of the groove and at the sealed distal end and (ii) the current through the system in sealed and open configuration. We have found that the spread of voltage is in quantitative agreement with the stationary solutions of Kelvin's equation for a homogeneous cable.     

Host defense mechanisms of cephalopods

Humoral and cellular mechanisms of defense have been described for cephalopods, a relatively advanced group of mollusks. Typical of other mollusks, cephalopod agglutinins are the most documented component of humoral immunity. Lectins, which have agglutinating properties, have been described and characterized from octopuses. Agglutinins from cephalopod hemolymph have also been shown to agglutinate a variety of vertebrate red blood cells, as well as potential bacterial pathogens. Hemocytes are the primary component of cellular immunity. Although the hemocyte role in phagocytosis has been extensively studied in other mollusks, the mechanisms of phagocytosis have not been described extensively for cephalopods. Cephalopod hemocytes have phagocytic capabilities and may function in encapsulation and neutralization of foreign substances; however, the effects of environmental factors and the full extent of phagocytic capabilities of cephalopod hemocytes have not been reported. Hemocytes from cephalopods have a role in wound healing and inflammation which have been reported in detail by several investigators.     

Biomass estimation of orange roughy: a summary and evaluation of techniques for measuring stock size of a deep-water fish species in New Zealand

A number of techniques have been employed to measure biomass of orange roughy in New Zealand. Time series of stratified random trawl surveys in a number of areas over periods of 5–10 years have given relative abundance indices, which have been used in stock reduction analyses to estimate true biomass. This has given confident results for the Chatham Rise stock, and surveys have also shown strong trends in other regions. Acoustic surveys have been carried out on three grounds. They can give relative indices of orange roughy abundance in areas of fiat or medium sloping bottom, and have the potential for estimation of absolute biomass from a single survey. Egg production surveys have been employed on two grounds on the east coast of the North Island, where conditions of tight spawning aggregations and steep bottom topography have limited the success of other methods. Both daily fecundity reduction and annual egg production methods have been used. These have given estimates of true biomass from one-off surveys, although results are imprecise. Both unstandardized and standardized analyses of commercial catch-per-unit-effort data have given relative indices of abundance, which have formed an important part of stock assessment for several fisheries. No single technique used to measure the size of orange roughy stocks has proven ideal or appropriate in all New Zealand situations. All have advantages and disadvantages, depending on the characteristics of the fishing area and fish behaviour. For two areas, a combination of methods have been applied, which has given more confident results than those from a single technique.     

CD4 cell surface downregulation in HIV-1 Nef transgenic mice is a consequence of intracellular sequestration.

The Nef gene product is a regulatory protein of HIV whose biological function is poorly understood. Nef has been thought to have a negative effect on viral replication in vitro but has been shown in studies with SIV to be necessary in the establishment of viraemia in vivo. In vitro studies in various human cell lines have shown that Nef downregulates the expression of cell surface CD4 and thus could have effects on the immune response. We have generated four transgenic mouse lines, with constructs containing two different Nef alleles under the control of CD2 regulatory elements to examine the interaction of Nef with the host immune system in vivo. In adult transgenic mice we have found marked downregulation in the level of CD4 on the surface of double positive thymocytes and a decrease in the number of CD4+ T cells in the thymus. Functional analyses have revealed a decrease in the total activation of transgenic thymocytes by anti-CD3 epsilon antibody. By specific intracellular staining of T cells in such mice we have found CD4 colocalizing with a Golgi-specific marker. These results strongly suggest a Nef mediated effect on developing CD4 thymocytes resulting from interference of Nef in the intracellular trafficking or post-translational modification of CD4.     

Pathogenesis of neonatal group B streptococcal infections

Infections of the neonate due to the group B Streptococcus have been recognized since the 1930s, but it was during the 1970s that their incidence grew alarmingly throughout the world. A research effort stimulated by this problem has yielded significant new information about many facets of the pathogenesis of these infections. Immunologic investigations have pinpointed a lack of transplacentally acquired antibody as a significant risk factor. In the laboratory, assays of antibody which have a functional endpoint have demonstrated that the type-specific carbohydrate antigens play a major role in stimulating the development of protective antibody. These assays have been shown to correlate with certain tests of primary antigen-antibody interaction which do not have a functional endpoint, but are simpler to use in larger scale epidemiologic studies. These tools may be useful in filling the gaps in our current knowledge of the pathogenesis of this infection.     

Phylogenetic analysis of the distribution of chromosomal races of Mus mnscuius domesticus Rutty in Europe

Robertsonian (Rb) translocation is a common chromosomal rearrangement in the house mouse. In free-living populations, 79 fusions with different combinations of chromosomes 1 to 18 have been found in some 45 populations. An updated list of these fusions is presented and analysed in order to reveal the possible processes by which the fusions spread within or among populations. A widespread hypothesis is that when two populations share the same fusion, it can be assumed that they have a common ancestor. This can serve as the basis for the use of the cladistic methods. While I present such an analysis on the updated list of Rbs, I also point to the problems associated with it in this case because many fusions have multiple origins and exchanges of Rbs between populations are frequent. I have tried to use a different approach, based on a critical and quantitative evaluation of the hypothesis of common ancestry. Assuming that the 153 possible fusions have an equal probability of occurrence, I give the formula to compute the probability that populations share a given number of fusions by chance alone. Only when this probability is lower than a chosen level (say 5%) can the populations be inferred to have a non-independent origin (i.e. they have a common ancestor or they have exchanged chromosomes by introgression). This probability measure is then used as a distance estimate to show the relationship between all the Rb populations. This analysis suggests that although some Rbs must have occurred more than once, most of the populations have non-independent origins. Almost all the populations from northern Africa to Belgium and Germany appear to have close karyotopic relationships and form a major group. Clearly independent Rb populations are mainly found in the periphery of this major group, for example in Scotland, Denmark and Spain. 'Chromosomal' flow between Rb populations appears to be a very important process.     

Identification and Functional Expression of HCx31.9, a Novel Gap Junction Gene

By combining in silico and bench molecular biology methods we have identified a novel human gap junction gene that encodes a protein designated HCx31.9. We have determined its human chromosomal location and gene structure, and we have identified a putative mouse ortholog, mCx30.2. We have observed the presence of HCx31.9 in human cerebral cortex, liver, heart, spleen, lung, and kidney and the presence of mCx30.2 in mouse cerebral cortex, liver and lung. Moreover, preliminary data on the electrophysiological properties of HCx31.9 have been obtained by functional expression in paired Xenopus oocytes and in transfected N2A cells.     

Identification and functional expression of HCx31.9, a novel gap junction gene

By combining in silico and bench molecular biology methods we have identified a novel human gap junction gene that encodes a protein designated HCx31.9. We have determined its human chromosomal location and gene structure, and we have identified a putative mouse ortholog, mCx30.2. We have observed the presence of HCx31.9 in human cerebral cortex, liver, heart, spleen, lung, and kidney and the presence of mCx30.2 in mouse cerebral cortex, liver and lung. Moreover, preliminary data on the electrophysiological properties of HCx31.9 have been obtained by functional expression in paired Xenopus oocytes and in transfected N2A cells.     

植物硫转运蛋白研究进展

硫转运蛋白在植物对硫酸盐的吸收和转运中起着重要的作用。已经在拟南芥、大麦和小麦等植物中分离到了40多种硫转运蛋白基因。这些基因序列与其他种类生物的硫转运蛋白基因序列有着高度的保守性。利用CLUSTAL程序建立的系统进化树将植物硫转运蛋白划分为5个亚群。使用多种拓扑预测程序推测出不同植物硫转运蛋白的共同结构特点是均含有12个跨膜域。在柱花草和大麦中,硫转运蛋白基因表达调控包括植物体内硫水平的负调控和O—乙酰丝氨酸的正调控两种方式。对硫转运蛋白的组织定位和功能研究表明,高亲和硫转运蛋白主要定位于根部,在根系硫酸盐吸收中起重要作用。     

Microbes, warfare, religion, and human institutions

A significant number of practicing microbiologists are not aware of the historical impact of infectious agents on the development of human institutions. Microbes have played a profound role in warfare, religion, migration of populations, art, and in diplomacy. Boundaries of nations have changed as a result of microbial diseases. Infectious agents have terminated some kingdoms and elevated others. There is a need for microbiologists to have a historical perspective of some of the major ways in which a pathogen may influence civilized populations. Conditions may exist in contemporary society for a repeat of some of the kinds of plagues suffered by previous societies. The purpose of this paper is to review examples of situations where pathogenic microbes have forced societal modifications on centers of human population.     

Microelectrodes for studying neurobiology

Microelectrodes have emerged as an important tool used by scientists to study biological changes in the brain and in single cells. This review briefly summarizes the ways in which microelectrodes as chemical sensors have furthered the field of neurobiology by reporting on changes that occur on the subsecond time scale. Microelectrodes have been used in a variety of fields including their use by electrophysiologists to characterize neuronal action potentials and develop neural prosthetics. Here we restrict our review to microelectrodes that have been used as chemical sensors. They have played a major role in many important neurobiological findings.     

Vegetation changes in the Pilliga forests: a preliminary evaluation of the evidence

Changes in the vegetation of Australia since white settlement have been much discussed in recent times. In particular, the changes that have been reported to have occurred in the Pilliga forests in northern New South Wales have been used as a reference for other areas of the State. Two periods of pine regeneration are believed to have occurred in the Pilliga, but preliminary research concerning the history of these forests has uncovered various sources of information indicating that the story is a more complex. Climatic data, archival records and the biology and ecology of various flora and fauna are examined in this paper in a preliminary attempt to gain a more accurate picture of change or stability in the vegetation of this region.     

Design and synthesis of mono- and multimeric targeted radiopharmaceuticals based on novel cyclen ligands coupled to anti-MUC1 aptamers for the diagnostic imaging and targeted radiotherapy of cancer

Targeted radiopharmaceuticals offer the possibility of improved tumor imaging and radiotherapy, with reduced side effects. A variety of monoclonal antibodies and antibody fragments have previously been successfully radiolabeled and used in diagnostic imaging and targeted radiotherapy of cancer. Many such antibodies have been shown to recognize the well-characterized MUC1 tumor marker and have recently been in clinical trials. Furthermore, a number of chelators have been synthesized and are currently used as radiopharmaceuticals for imaging and therapy. We now report the synthesis of a novel, cyclen-based ligand with a sulfur-containing arm that offers increased stability of the ligand-metal complex. We have coupled this ligand with previously selected aptamers to the MUC1 tumor marker to generate a novel targeted radiopharmaceutical with improved properties. We have tested the complex against known, commercially available chelators such as MAG3 in model breast cancer systems. To improve the pharmacokinetic properties of the aptamer-based targeted radiopharmaceutical, we have generated multi-aptamer complexes around a central chelator. Such multi-aptamer complexes have increased retention of the complex in circulation, without affecting the lack of immunogenicity of the complex or altering its superior tumor penetration properties.     

Biodiversity among haptophyte algae

The division Haptophyta is represented only by about 300 extant species showing wide diversity in morphology, biochemistry and ecology. They have a world-wide distribution and are numerically important in phytoplankton populations in nearly all marine environments. Evidence from the geological record shows that they have been the major constituent of calcareous deposits since the Late Triassic and, as they have evolved quickly through time, their coccoliths have always shown wide morphological diversity. In today's oceans they occasionally produce extensive blooms, visible by satellite imagery, which have ecological impact. As a consequence of these blooms the haptophyte algae are now receiving greater attention, as their role in the global sulphur and carbon cycles may influence the world's climate, and their potential as nuisance bloom algae have implications for commercial fishing and the marine ecosystem. As it is likely that these organisms have always produced such blooms, these effects may have been in operation for the last 200 million years.     

Chromium in carbohydrate and lipid metabolism

 Since the discovery in the 1950s that mammals have a nutritional requirement for chromium, the biological function of chromium has been sought. Candidates for the naturally-occurring biologically active form of chromium have been proposed, but, until recently, all have been shown to be artifacts. Recent studies examining the properties of the oligopeptide low-molecular-weight chromium-binding substance (LMWCr) suggest that this material may have a role in carbohydrate and lipid metabolism as part of a novel insulin-signaling amplification mechanism and may have implications in the treatment of diabetes and related conditions. Received: 24 March 1997 / Accepted: 9 September 1997