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.     

Studies on the proteins of Poecilocera picta. I-changes taking place in the haemolymph protein during moulting

The haemolymph proteins of the 6th nymph of P. picta were fractioned by the polyacrylamide gel disc electrophoresis. A total of seven proteins fractions have been detected from the haemolymph. The chemical nature of different protein fractions have been examined by histochemical methods. The changes taking place in the cuticle and epidermal cells have been examined during the transformation of 6th nymph into adult. The fat body proteins have been electrophoretically fractioned and the changes in the concentration of different protein fractions have been examined. It is suggested that the protein fraction 3 of the haemolymph is utilized in the formation of new cuticle. It is concluded by the histochemical observations that proteins of the band 3 are synthesized in the fat body.     

Advances and prospects: biotechnologically improving crop water use efficiency

Bio-water saving can be defined as the reduction of crop water consumption employing biological measures. This is the focus of efforts to save water in agriculture. Different levels of water-use efficiency (WUE) have been developed. The genetic diversity of WUE has been confirmed in several crops. WUE is the basis of bio-watering and physiological WUE is the key. The degree to develop physiological WUE potential decides the performance of bio-watering in the field. During this process, fine management is important. Thus bio-watering is closely related to WUE. Crop WUE has improved and evolved as a result of breeding programs. Many WUE genes have been located in different genomic and aneuploid materials and have been mapped by various molecular markers in a number of crops. Two genes, (Erecta and alx8), which control water use efficiency; have been cloned in Arabidopsis thaliana. Eleven WUE genes have been identified by microarray analysis. Six genes associated with drought resistance and photosynthesis have been transfered into crops which have resulted in improving WUE and drought resistance. WUE is important on the basis of functional identification of more drought resistant gene resources. The popularity on the industrial-scale of transgenic plants is still in its infancy and one of the reasons for this is the lack of knowledge regarding molecular mechanisms and it is a very immature technology. Enhanced agricultural practices and the theoretical aspects of improving crop WUE have been developed and are discussed in this review paper. Rapid progress will be made in bio-water savings and that crop WUE can be substantially improved under both favorable and unfavorable water-limited environments. This will be achieved by a combination of traditional breeding techniques and the introduction of modern biotechnology.     

Development of the genetic map of the yeast Saccharomycopsis lipolytica

Summary Tetrad and random spore analyses have been used to further develop the genetic map of Saccharomycopsis lipolytica. Mutations in 23 new nuclear genes have been isolated. Eight genes have been located on linkage fragment 1, 4 on fragment 2, 2 on fragment 5 and 3 on fragment 6. Linkage fragments 3 and 4 have been shown to be linked, and this fragment now contains 12 markers. A tentative map of the linkage fragments 1 and 3 is presented (Fig. 1). Markers exhibiting possible centromere linkage have been identified. Interference estimates suggest that there is little interference in S. lipolytica.     

Polymorphism in the Aegean "five-loculed" species of the genus Campanula, Section Quinqueloculares (Campanulaceae)

Population variation in species of Campanula Section Quinqueloculares and related taxa has been studied. 19 species and 48 populations from the Aegean region have been sampled. There are few distinct character states available for analysis so that metric variables have been scored. 11 leaf variables and 19 flower variables have been measured in up to 683 plants. Regressions of variables with overall size have been calculated and where significant allometric variation has been detected residual scores have been used as variables. Hierarchical analysis of variance has indicated that a large part of the total variation is found between populations within species rather than within populations. Between the 10 most closely related species there is a considerable overlap in population means of variables. Multivariate analysis also fails to identify distinct species clusters. In particular principal components analysis provides additional evidence of overlap between species. It is suggested that a detailed study of genetic variation within and between species is required in order to establish the presence of any distinct phylogenetic lineages.     

Biotransformations using prokaryotic P450 monooxygenases

Recent studies on microbial cytochrome P450 enzymes have covered several new areas. Advances have been made in structure-function analysis and new non-enzymatic/electrochemical systems for the replacement of NAD(P)H in biocatalysis have been developed. Furthermore, the properties of some enzymes have been re-engineered by site-directed mutagenesis or by methods of directed evolution and new P450s have been functionally expressed and characterized. It is thought that a combination of these approaches will facilitate the use of isolated P450 monooxygenases in biocatalysis.     

Morphologie,ontogénie et valeur adaptative des surfaces glandulaires mucilagènes dans le genre Chenopodium L (Chenopodiaceae)

The abaxial and adaxial epidermis of young and mature leaves of the South American species of Chenopodium L. (Chenopodiaceae) have mucilage secreting glandular structures that have not been reported in previous studies, either for the Chenopodiaceae or other Phanerogamic taxa. Their morphology and function have been studied and all stages of their ontogenic pathway have been described and illustrated. The name ‘glandular mucilagene surface’ is proposed. Their adaptative value is discussed and differences from other leaf epidermal differentiations are noted. These structures have been discovered in Chenopodium species living in Patagonia, their presence in all South American species — native and exotic ones — of the genus has been investigated and proved. The possibility that it could be a generic feature is suggested.     

"Enzymatic" lipid peroxidation: reactions of mammalian lipoxygenases

Lipoxygenase is a dioxygenase which incorporates one molecule of oxygen at a certain position of unsaturated fatty acids such as arachidonic and linolenic acids. The enzymatic oxygenation of unsaturated fatty acids is stereospecific concomitant with a stereoselective abstraction of hydrogen atom. Fatty acid cyclooxygenase is an atypical lipoxygenase incorporating two molecules of oxygen, and initiates the biosynthesis of prostaglandins and thromboxanes. Arachidonate 5-lipoxygenase is responsible for the leukotriene synthesis. No such bioactive compound has been found as a metabolite of the 12- and 15-lipoxygenase pathways, and their physiological roles are still unclarified. These enzymes have been purified, and their molecular and catalytic properties have been investigated. Their cDNA clones have been isolated, and their nucleotide sequences have been determined deducing the primary structures of the enzymes.     

THE MOLECULAR EVOLUTION OF PITUITARY HORMONES

1. The pituitary hormones can be divided into 4 families; within each the members are structurally related and have probably evolved from a common ancestor by a process of gene duplication and divergence. 2. Recent structural studies have revealed much about the evolution of proteins. The roles of point mutation, gene duplication and partial gene duplication in molecular evolution have been highlighted, and the nature of the evolutionary forces involved has been extensively debated. The information available about the evolution of proteins in general provides a background for consideration of pituitary hormone evolution. 3. The structure and function of the mammalian neurohypophysial hormones (oxytocin and vasopressin) has been studied in detail. Related (structurally similar) peptides have been found in the neurohypophyses of lower vertebrates and have been Characterized in many instances. Several schemes have been proposed for the evolution of these hormones. 4. The vasopressins of the pig and its relatives show a genetic polymorphism. The roles of neurohypophysial hormones in lower vertebrates are very varied and not fully understood. 5. The ACTHs and MSHs are members of a second family of pituitary hormones. They are polypeptides of moderate size. Studies on amino-acid sequences have been carried out for ACTHs and MSHs from several mammals. α-MSH is identical in all cases studied in detail, but β-MSH and ACTH vary to some extent. There is considerable sequence homology between the hormones in this family - indicating a common phylogenetic origin and several gene duplications. 6. Dogfish MSH is the only non-mammalian hormone of the ACTH-MSH family to have been studied in detail. Two MSHs have been isolated from this species; both resemble the a-MSH of mammals in amino-acid sequence. ACTH-like and MSH-like hormones exist in many other vertebrate groups, but have not been characterized fully. 7. Structure-function relationships have been widely studied in the ACTH-MSH family, and have some interesting evolutionary implications. Polymorphism of P-MSHs is found in some mammals. 8. A third family of protein hormones includes pituitary prolactin and growth hormone, and placental lactogen. These are proteins of moderate size which have been shown to be widely distributed among the vertebrates. Species specificity can be recognized with regard to biological, immunological and structural properties. 9. Amino-acid sequences have been determined for growth hormones and prolactins from several mammals. There is sequence homology between growth hormone and prolactin. Human placental lactogen closely resembles human growth hormone. A phylogenetic tree has been constructed for this protein family. Rates of evolution within the group are rather variable. 10. The fourth family of pituitary hormones (FSH, LH, TSH and some related placental hormones) are all glycoproteins and have a subunit structure. Extensive sequence studies have been carried out on the hormones from some mammals, and show that there is considerable homology between the various subunits. The α-subunits of human TSH, LH and HCG (and probably FSH) are very similar. The β-subunits are different, but homologous. Evolution of this family clearly took place by a series of gene duplications followed by gene divergence. Schemes whereby this could have occurred have been discussed. Related hormones occur in lower vertebrates, but have not been fully characterized. Some lower vertebrates may possess only one gonado-trophin. 11. The pituitary hormones provide an interesting range of evolutionary problems, and are useful models for the study of molecular evolution. The evolutionary processes involved in their diversification have been discussed, with particular reference to the co-evolution of hormones and their receptors. Neutral mutations and gene duplications may have played a role in providing co-existing variation of hormones and receptors. 12. A speculative model for the evolution of neurohypophysial hormones is proposed, as an example of how molecular evolution may have operated in this and other hormone groups. 13. Homologies have been proposed between the various families of pituitary hormones, and between pituitary proteins and other entero-secretory proteins. The pituitary protein hormones were probably elaborated from smaller molecules by a process of partial gene duplication.     

THE ACCUMULATION OF ELECTROLYTES : VII. ORGANIC ELECTROLYTES PART 2

Analyses have been made of the inorganic constituents of the juices expressed from the leaves of Rheum, Rumex, and Oxalis. It has been shown that in all cases there is a large excess of inorganic cations over anions in the sap, the average ratio of cations to anions being 3.8 (Part 1, p. 239). The ash analyses of plant tissues (chiefly leaves) reported in the literature have been examined critically, and it has been shown that the preponderance of inorganic cations over inorganic anions in the ash and in the sap is general. It has been concluded that the excess of inorganic cations is consistent with the view that cations pass into the protoplasm chiefly in the form of hydroxides, and are accumulated either in the form of organic salts (such as the oxalates) or in non-polar linkage. It has been concluded that practically all the potassium and sodium found in plant ash must have been present originally in the form of soluble ionogenic compounds, but that a considerable part of the calcium and magnesium may have been present originally in the form of insoluble salts or as components of non-polar compounds. The methods whereby the cations, particularly potassium, may have been accumulated have been discussed, and it has been concluded that as it does not seem very probable that they enter chiefly as nitrates or bicarbonates we may suppose that they go in to a large extent as hydrates: this is highly probable in the case which has been most carefully investigated (Valonia).     

水龙骨科植物的引种及种质资源保存

通过对５０种水龙骨科（Ｐｏｌｙｐｏｄｉａｃｅａｅ）植物的引种栽培,研究了水龙骨科植物的引种方法、栽培管理措施和繁殖技术,初步应用离体保存技术保存了１０种水龙骨科植物,并对主要类群做了应用及观赏性状评价。     

A chromosomal walk in the region of polytene bands 7C-D of theDrosophila X chromosome

A chromosomal walk on the X chromosome ofDrosophila in the region of polytene bands 7C1 to 7D5 is described. The region is of interest since three olfactory genes have been found to map here in addition to a haplo-inviable locus. Genomic clones spanning 160 kilobases have been isolated and their complete restriction map is presented. The clones have been aligned on the polytene chromosome bands byin situ hybridisation. In addition the end-points of a deficiency and duplication lying in this region have been mapped approximately, showing that an overlap exists between them.     

Physiological effects of dominance hierarchies: laboratory artefacts or natural phenomena?

Studies of fish behaviour have demonstrated the existence of social interactions that result in dominance hierarchies. In environments in which resources, such as food, shelter and mates, are limited, social competition results in some fish becoming dominant and occupying the most profitable positions. This behaviour has been observed in natural environments and also in many laboratory‐based experiments. When two fish have been confined in a small tank, one of them usually has exhibited behaviour that suggests it is dominant over the other submissive animal. Physiological consequences of social interaction can be seen in both dominants and subordinates but are more extreme in the subordinate. However, this scenario is without doubt an artificial situation. Fewer experiments have been conducted using laboratory experiments that are more socially and physically complex than those experienced by dyads in tanks. In simple fluvial tanks, through which water is recirculated, the physiological responses of fish to social competition have generally been qualitatively similar to those recorded among dyads. However, when environmental disturbances, complex resource distributions, increase in water flushing, presence of predators and competing species of fish have been included in experimen‐tal designs, there have been fewer, diminished or no physiological dierences between dominant and subordinate fish. There have been very few studies of physiology in relation to dominance in natural habitats, and those that have been conducted suggest that under some circumstances hierarchies may cause less intense physiological responses than have been suggested based on results of laboratory studies in simple environments. Possible reasons for these variations are discussed. The need is identified for a well structured experimental approach to the investi‐gation of the causes and consequences of hierarchies if the ecology of wild fish is to be modelled eectively based on physiological processes. It is also suggested that the further development and application of techniques for monitoring physiologies of fish in the wild is important.     

In vitro compression of a soft tissue layer on a rigid foundation

In vitro compression studies have been performed on layers of porcine skin and fat. The tissue layers have been loaded by means of various indentors. Indentor displacements and interstitial fluid pressures have been measured. The results have been compared to finite element calculations with mixture elements. A qualitative agreement between calculations and measurements is found. The results support the hypothesis that skin and fat behave like solid/fluid mixtures.     

Insect neuronal cultures: an experimental vehicle for studies of physiology, pharmacology and cell interactions

The current status of insect neuronal cultures is discussed and their contribution to our understanding of the insect nervous system is explored. Neuronal cultures have been developed from a wide range of insect species and from all developmental stages. These have been used to study the morphological development of insect neurones and some of the extrinsic factors that affect this process. In addition, they have been used to investigate the physiology of sodium, potassium and calcium channels and the pharmacology of acetylcholine and GABA receptors. Insect neurones have also been grown in culture with muscle and glial cells to study cell interactions.     

The elusive sizer

Size control has been a topic of interest to cell biologists for over a century, but insights into cell size control mechanisms have until recently been relatively sparse. Determining whether cells have a size measurement mechanism and how it might operate has proven difficult. The nucleocytoplasmic ratio is one of the few conserved features of size control but little is know about how it is measured. Models where growth and division can be uncoupled have been underexploited, but have considerable potential for gaining insights into the contribution of the nucleocytoplasmic ratio to cell size regulation.     

Sensitivity of parasites to free radical damage by antiparasitic drugs

Over the last few years a remarkable progress has been made in the understanding of parasites biochemistry, molecular biology, and immunology. This progress is especially encouraging in that emphasis on drug development is shifting from random screening towards a more rational approach. A number of peculiar aspects characteristic of parasites which are not present in other organisms and that might be exploitable for the design of specific agents have been described recently. One of these aspects is their deficiency in defense mechanisms against oxygen toxicity. Catalase is absent in many parasites. Distinct superoxide dismutases have been detected and specific inhibitors of these enzymes have been investigated. Glutathione is absent in some anaerobic protozoa. Peroxidase and reductase activities dependent on a glutathione-spermidine cofactor termed trypanothione have been detected in several trypanosomatids and apparently replace the glutathione peroxidase-glutathione reductase system of other eukaryotic cells. Free radical intermediates have been shown to be involved in the reaction of enzymes present in anaerobic protozoa. In addition, a number of antiparasitic agents have been shown to exert their actions through a free radical metabolism: nitro compounds used against trypanosomatids, anaerobic protozoa and helminths; crystal violet used in blood banks to prevent blood transmission of Chagas' disease; the antimalarial primaquine, chloroquinine, and quinhasou; and quinones active in vitro and in vivo against different parasites.     

Histologic organization of the myocardium of the rat during hypokinesia (stereologic study)

For 30 days Wistar rats have been kept under conditions of hypokinesia. Using stereological methods, the volume and surface density of muscle fibers, microcirculatory bed and connective tissue components have been defined, and volumetric and surface--volume ratios of parenchymal and stromal structures have been calculated. Intracellular stereological parameters of the cardiac myocytes have been studied. Three-dimensional parameters in the parenchymatous-stromal interrelations of the atrophying myocardium accompanied with a deep rearrangement of the intracellular architectonics have been determined, as well as certain destructive changes of the cardiac myocyte ultrastructures. Such a treatment as a decreased functional loading is demonstrated to be the cause of cardiomyocytic lesions.     

Detection of diuretic agents in doping control

Since the inclusion of diuretics in the list of banned substances in sports in 1988, a large number of screening and confirmation procedures to detect the presence of these substances in urine samples have been developed. In this paper, a review of the analytcal methodology described to analyze diuretics is presented. The paper has been focused on the needs of doping control and mainly screening procedures including sample preparation and liquid or gas chromatographic separation have been considered. More relevant papers using capillary zone electrophoresis have been also considered. Mass spectrometry is mandatory in doping control for confirmation purposes, and finally, mass spectrometric techniques described for diuretics have been reviewed.     

Changes in the geographical distribution of malaria throughout history

Climatic changes must have greatly affected the distribution of malaria in prehistoric times. Paleobotanical evidence, snowline depression studies and information obtained from deep sea sediment cores, indicate that southern Europe must have suffered a drop of summer temperatures of approximately 9 degrees C during the last glacial maximum, 18,000 years ago. Such a drop would have been decisive as regards the distribution of malaria and its vectors. If present at all, the disease would have been confined to the southernmost parts of the continent but P. falciparum and today's most effective vectors--A. labranchiae and A. sacharovi--would have been excluded from Europe. In western Asia, summer temperatures 6 degrees C lower than those of today would have had less effect on the malaria situation. The introduction of falciparum malaria in southern Europe is placed in Hellenistic and Early Imperial Roman times, based on paleoclimatological evidence and historical and medical data. In America P. falciparum is also considered a late entrant but vivax and quartan malaria may have been introduced in pre-Columbian times. In the Pacific, the disease is known to have been spread by man since the Age of Discovery until contemporary times.