Does the tube of a benthic chironomid larva play a role in protecting its dweller against chemical toxicants?

A laboratory culture of an Israeli benthic midge, Chironomus luridus, was exposed to two chemicals: copper and monochloramine. The objective of this study was to determine the protective nature of Chironomus' larval tube. Three experimental conditions were tested: larva with sand tubes, with silt tubes and without tubes. Larvae without tubes were significantly more sensitive to copper and chloramine than larvae that had sand or silt as tube building substrate. The tubes protected the insects against chemicals throughout 14 days of exposure time. Silt tubes had higher protective value than sand tubes, especially when exposed to copper for a short period of time (LC₅₀/ 24 h, with silt, sand, or none: 80.0, 7.0 and 3.4 mg l^–1 copper, respectively). C. luridus seemed to be better protected against copper than against chloramine (LC₅₀/ 24 h, with silt, sand, or none: 12.2, 6.4 and 3.7 mg l^–1 chloramine, respectively). The acute toxicity of copper to chironomid larvae was investigated using a cytochemical method. Larva in silt tubes had significantly higher non-specific esterase activity in midgut cells than larvae without tubes. We conclude that, in addition to its role in feeding, respiration and anti-predation shelter, the C. luridus tube protects its inhabitant from toxic substances.     

Enhanced Levels of the Aroma and Flavor Compound S-Linalool by Metabolic Engineering of the Terpenoid Pathway in Tomato Fruits

The aromas of fruits, vegetables, and flowers are mixtures of volatile metabolites, often present in parts per billion levels or less. We show here that tomato (Lycopersicon esculentum Mill.) plants transgenic for a heterologous Clarkia breweri S-linalool synthase (LIS) gene, under the control of the tomato late-ripening-specific E8 promoter, synthesize and accumulate S-linalool and 8-hydroxylinalool in ripening fruits. Apart from the difference in volatiles, no other phenotypic alterations were noted, including the levels of other terpenoids such as gamma- and alpha-tocopherols, lycopene, beta-carotene, and lutein. Our studies indicate that it is possible to enhance the levels of monoterpenes in ripening fruits by metabolic engineering.     

Senescence-associated mRNAs that may participate in signal transduction and protein trafficking

The differential display technique was used to generate cDNA probes in order to identify mRNAs that are up-regulated during senescence of Arabidopsis leaves. Three mRNAs were examined that had not previously been associated with senescence. The steady-state levels of these mRNAs are detectable in small amounts in mature green leaves, but increase considerably as chlorophyll levels begin to decline. This relationship to senescence occurs under natural circumstances as well as when senescence is accelerated by leaf detachment in the dark or by addition of 1-aminocyclopropane-1-carboxylic acid (ACC). Retardation of senescence by benzyladenine slows the increase of the mRNAs. One of these mRNAs appears to code for a protein (Sec 13) that may be involved in vesicle formation at the endoplasmic reticulum. Another mRNA codes for a protein with WD‐repeat motif whose function is as yet unidentified, and the third codes for a putative calcium-dependent protein kinase. A fourth cDNA has also been cloned by subtractive hybridization from senescing Arabidopsis leaves that encodes vacuolar-processing enzyme ( γ VPE). Incubation of detached leaves in darkness also caused an abrupt elevation in the steady-state levels of the γVPE , similar to that of the senescing attached leaves. The possible functions of the gene products and their involvement in cellular and biochemical processes during senescence are discussed.     

The role of a spiny plant refuge in structuring grazed shortgrass steppe plant communities

This study evaluates the hypothesis that biological grazing refuges have an important role in plant-grazer interactions of grasslands with a long history of grazing. We assessed the hypothesis that clumps of the spiny cactus Opuntia polyacantha provide biological refuges from cattle grazing, affecting cover and seedhead production of associated vascular plants in the shortgrass steppe of the North America. The study was based on sampling inside and outside Opuntia clumps in eight long-term moderately grazed pastures established 60 yr ago and their respective ungrazed controls. Opuntia clumps provided a refuge for seedhead production of the dominant grass ( Bouteloua gracilis ) and for cover and seedhead production of many plant groups. Clumps were also a refuge for species sensitive to grazing (species that decrease with grazing) and barrel-cacti, but not for species preferred by cattle (species with greater proportion in the diet than in the field), exotics or weeds. Our results suggest that these effects were mainly through changes in the microenvironmental conditions resulting from protection effects, even though all potential microenvironmental effects could not be measured. Cacti promoted some negative effects on other plant groups, probably due to the space occupied by cladodes inside cactus clumps. The refuge effects observed at the group level did not translate into strong community level effects. Species diversity ( H' ) was greater in cactus clumps due to lower dominance rather than greater richness. The presence of Opuntia clumps increased landscape-scale diversity. This ecological role of Opuntia clumps as refuge from cattle grazing should be taken into consideration in management practices aimed at cactus eradication in order to increase forage availability for livestock. We discuss the potential role of plant community productivity and grazing history with regard to the importance of natural refuges in structuring grassland communities.     

An in situ study of beta-glucosidase activity in normal and gaucher fibroblasts with fluorogenic probes

Beta-glucosidase activity was evaluated in situ by means of fluorogenic probes in normal human fibroblasts and fibroblasts from homozygous carriers of the Gaucher trait. Probe internalization, targeting to lysosomes and post-cleavage probe retention were the primary concerns. Internalization and targeting were attempted by in situ photosensitized labilization of lysosomal membranes, lysosomotropic detergents and the use of low density lipid (LDL) or the receptor ligand apolipoprotein E (ApoE). Post-cleavage increase of fluorescence with fluoresceinyl (bis) betaglucopyranoside was appreciably above the rather large pre-cleavage emission. In cells incubated overnight with nonylumbelliferylbetaglucoside (UG9) in the presence of bovine serum albumin and in the absence of ApoE, the probe was dealt with as a cytotoxic agent, accumulating in a paranuclear cap, most likely comprising elements of the endoplasmic reticulum (ER) and Golgi apparatus. Targeting of UG9 to lysosomes occurred within 1 to 3 h of preincubation in the presence of ApoE. There was some evidence of specificity, as Gaucher fibroblasts exhibited weaker cleavage of UG9 (by 50 per cent or more) compared to normal fibroblasts, but in the Gaucher cells there was some residual beta-glucosidase activity. Cleavage of UG9 was nearly totally suppressed in Gaucher cells treated with the beta-glucosidase inhibitor, conduritol B epoxide, for 24 h to 7 days. Suppression in the control fibroblasts was evident but to a lesser degree. The in situ method of fluorogenic assay established for beta-glucosidase deficiency, is in principle applicable to enzyme deficiencies in other lysosomal storage diseases, or to evaluate enhanced enzyme activity following gene therapy.     

Mechanism of activation of Sepharose and Sephadex by cyanogen bromide

The mechanism of CNBr activation of polysaccharide resins like Sepharose and Sephadex has been elucidated using recently published analytical procedures for the determination of cyanate esters and imido carbonates. It was found that on agarose-based resins coupling of ligand occurs predominantly via cyanate esters, and not via imidocarbonates as in the case of Sephadex. This explains the different behaviours of Sepharose and Sephadex during CNBr activation.     

The effect of implanted, active corpora allata on reproductive diapause in adult females of the grasshopper Oedipoda miniata

The effect of implanted active corpora allata (CA) on the reproductive diapause in adult females of the grasshopper, Oedipoda miniata, was investigated. Implantation of two pairs of CA originating from sexually mature, adult male Locusta migratoria migratorioides donors effectively terminated reproductive diapause by inducing oöcyte development and intensive egg-laying in O. miniata recipients at any state of the reproductive diapause. The interval elapsing from implantation till oviposition was 15 days when recipients were at the beginning of the reproductive diapause. This interval shortened when recipients were in later stages of the reproductive diapause, thus, closer to its eventual termination. Reproductive diapause in female O. miniata, therefore, seems to be a slow process rather than a steady state. In all instances recipients laid eggs for quite short periods (up to 3 weeks). After cessation of egg-laying resorption of the proximal oöcytes occurred in the ovarioles. The implanted glands presumably became inactive.Implantation of two pairs of CA originating from sexually mature (already in post-diapause state) male O. miniata donors also induced oöcyte development and occasional egg-laying in female recipients which were still in reproductive diapause at the time of implantation. The effect, however, was markedly less intensive than that induced by heterospecific implantation. Practically no oöcyte development was obtained in sham-operated and unoperated controls.

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Résumé L'implantation dans des femelles d'Oedipoda miniata en diapause génitale, de 2 paires de corpora allata (CA) prélevés sur des mâles adultes sexuellement mûrs de Locusta migratoria migratorioides, provoque la rupture de cette diapause, quel que soit son état d'évolution, en induisant le développement des ovocytes et une intense activité de ponte. La ponte est déclenchée dans un délai de 15 jours après l'implantation quand l'insecte receveur est en début de diapause génitale. Ce délai est plus court quand la diapause est déjà avancée, donc quand la greffe intervient à un moment plus proche de son achèvement normal. Par conséquent la diapause génitale chez la femelle d'Oedipoda miniata semble être un processus à évolution lente, plutôt qu'un état stable.Dans tous les cas les insectes ayant reçu un implant de CA pondent pendant une courte période (jusqu'à 3 semaines). Quand cesse la ponte la résorption des ovocytes proximaux survient dans les ovarioles; sans doute les glandes implantées sont devenues inactives.L'implantation de 2 paires de CA provenant de mâles d'Oedipoda miniata sexuellement mûrs (déjà dans un état de post-diapause) induit également le développement des ovocytes et occasionnellement la ponte chez des femelles en diapause génitale, mais l'effet est cependant nettement moins intense que celui induit par une greffe hétérospécifique. Pratiquement on n'observe aucun développement d'ovocytes chez les insectes témoins non-opérés ou ayant subi une pseudo-greffe.

     

Molecular Exploration of the First-Century Tomb of the Shroud in Akeldama,Jerusalem

The Tomb of the Shroud is a first-century C.E. tomb discovered in Akeldama, Jerusalem, Israel that had been illegally entered and looted. The investigation of this tomb by an interdisciplinary team of researchers began in 2000. More than twenty stone ossuaries for collecting human bones were found, along with textiles from a burial shroud, hair and skeletal remains. The research presented here focuses on genetic analysis of the bioarchaeological remains from the tomb using mitochondrial DNA to examine familial relationships of the individuals within the tomb and molecular screening for the presence of disease. There are three mitochondrial haplotypes shared between a number of the remains analyzed suggesting a possible family tomb. There were two pathogens genetically detected within the collection of osteological samples, these were Mycobacterium tuberculosis and Mycobacterium leprae. The Tomb of the Shroud is one of very few examples of a preserved shrouded human burial and the only example of a plaster sealed loculus with remains genetically confirmed to have belonged to a shrouded male individual that suffered from tuberculosis and leprosy dating to the first-century C.E. This is the earliest case of leprosy with a confirmed date in which M. leprae DNA was detected.