It is becoming clear that simple sequence repeats (SSRs) play a significant role in fungal genome organization, and they are a large source of genetic markers for population genetics and meiotic maps. We identified SSRs in the Laccaria bicolor genome by in silico survey and analyzed their distribution in the different genomic regions. We also compared the abundance and distribution of SSRs in L. bicolor with those of the following fungal genomes: Phanerochaete chrysosporium, Coprinopsis cinerea, Ustilago maydis, Cryptococcus neoformans, Aspergillus nidulans, Magnaporthe grisea, Neurospora crassa and Saccharomyces cerevisiae. Using the MISA computer program, we detected 277,062 SSRs in the L. bicolor genome representing 8% of the assembled genomic sequence. Among the analyzed basidiomycetes, L. bicolor exhibited the highest SSR density although no correlation between relative abundance and the genome ...
A monoclonal antibody raised against carboxyl-linked IAA was used in Western blot analysis of storage proteins from kernels of Avena sativa, Pennisetum americanum, Sorghum bicolor,...Full Text Available
... east Nigeria) and has also been recorded from south west Cameroon. Only two records; that from Angola the locality ... east Nigeria) and has also been recorded from south-west Cameroon. It is inferred tha...
Wild-type viruses from the ViroLogic phenotype-genotype database were evaluated to determine the upper confidence limit of the drug susceptibility distributions, or “biological cutoffs,”...Full Text Available
To analyze the proposed growth-inhibitory function of wild-type p53, we compared simian virus 40 (SV40) DNA replication in primary rhesus monkey kidney (PRK) cells, which express wild-type p53, and...Full Text Available
... including exotic species such as Hydrilla verticellata (hydrilla), Hygrophila polysperma (hygrophila), Cryptocoryne beckettii (water trumpet), leaving little space ... ...
The regulation of the glutamate dehydrogenases was investigated in wild-type Neurospora crassa and two classes of mutants altered in the assimilation of inorganic nitrogen, as either...Full Text Available
Background and AimsIt is well known that genome size differs among species. However, information on the variation and dynamics of genome size in wild populations and on the early...Full Text Available
Synthesis of wild-type Neurospora crassa assimilatory nitrate reductase is induced in the presence of nitrate ions and repressed in the presence of ammonium ions. Effects of several...Full Text Available
Behavioral responses to wind are thought to play a critical role in controlling the dispersal and population genetics of wild Drosophila species1,Full Text Available
... proficiency, most Army personnel must shoot to the same standards required of ... Wild- flecken was to provide this standard range, but its scores ... I3a I] ...
Newly hatched chicks were dosed orally with a Salmonella typhimurium wild-type strain, an S. enteritidis wild-type strain, and a genetically defined S. enteritidis aroA vaccine candidate, strain CVL30....Full Text Available
The relationship between plasma testosterone levels and locomotor activity in wild-caught sexually mature male meadow voles (Microtus pennsylvanicus) was assessed in the laboratory. Several aspects of locomotor activity were monitored for 1 h on two consecutive days using the automated Digiscan activity monitoring system. Plasma testosterone levels were determined immediately following the second day of activity monitoring. Significant Pearson correlations were obtained between plasma testosterone levels and total distance traveled [r(10) = 0.55, p < 0.05] and amount of time spent in movement [r(10) - 0.55, p < 0.05] on the second day. The wild voles showed a reduction in activity levels from the first to the second day of activity monitoring, which is indicative of habituation to a novel environment. This study provides direct evidence for a significant correlation between laboratory measures of behavioral activity and plasma ...
The existence of problems with wild pigs (Sus scrofa) is nothing new to the Western Hemisphere. Damage by these introduced animals was reported as far back as 1505 by the early Spanish colonies in the Caribbean, where wild pigs were killing the colonists cattle. Droves of these animals also ravaged cultivated crops of maize and sugarcane on islands in the West Indies during this same time period. These wild pigs reportedly were very aggressive and often attacked Spanish soldiers hunting rebellious Indians or escaped slaves on these islands, especially when these animals were cornered. The documentation of such impacts by introduced populations of this species in the United States has subsequently increased in recent years, and continued up through the present (Towne and Wentworth. 1950, Wood and Barrett 1979, Mayer and Brisbin 1991, Dickson et al. 2001). In spite of a fairly constant history in this country since the early ...
Abstract Host-microorganism interactions in the intestinal tract are complex, and little is known about specific nonpathogenic microbial factors triggering host responses in the gut. In this study, mannose-specific interactions of Lactobacillus plantarum 299v with jejunal epithelium were investigated using an in situ pig Small Intestinal Segment Perfusion model. The effects of L. plantarum 299v wild-type strain were compared with those of two corresponding mutant strains either lacking the gene encoding for the mannose-specific adhesin (msa) or sortase (srtA; responsible for anchoring of cell surface proteins like Msa to the cell wall). A slight enrichment of the wild-type strain associated with the intestinal surface could be observed after 8 h of perfusion when a mixture of wild-type and...
Objectives1. To create a strong, interdisciplinary evidence base about the relationships linking ecosystems, agrobiodiversity, wild biodiversity, and sustainable livelihoods, including the use of trade-off analysis (modeling the relationships between agricultural productivity, levels of biodiversity, and ecosystem resilience); crop evolutionary studies (wild relative/crop interactions) and socioeconomic, cultural, and nutritional studies of the contribution of biodiversity to different livelihoods asset [continued...]DescriptionAgriculture, ecosystems, and humans have co-evolved over millennia in the Andean-Amazonian region, creating the richest of all Vavilov centers of crop origin and diversity. The conservation of a wide range of domesticated crops and landraces (i.e., indigenous, ancestral varieties or cultivars that are distinct, uniform, and stable) and their co-evolution with crop wild relatives (CRW) has been ...
Virtually all biomedical research makes use of a relatively small pool of laboratory-adapted, inbred, isogenic stocks of mice. Although the advantages of these models are many, there are a number of...Full Text Available
Xylem vessels in grapevines Vitis labrusca L. and Vitis riparia Michx. growing in New England contained air over winter and yet filled with xylem sap and recovered...Full Text Available
Project ZEUS was designed to characterize the effects of long-term gamma irradiation on free-ranging meadow voles, and to determine the lowest level of radiation at which biological effects are discernible in the population. Behavioural tests are consider...
To evaluate the role of putative group A streptococcal virulence factors in the initiation of skin infections, we compared the adherence of a wild-type M49-protein skin-associated strain to that of...Full Text Available
Human cytomegalovirus (HCMV) in clinical material cannot replicate efficiently in vitro until it has adapted by mutation. Consequently, wild-type HCMV differ fundamentally from the passaged strains...Full Text Available
The effect of different nitrogen compounds on the induction of reduced nicotinamide adenine dinucleotide phosphate-nitrate reductase was examined in Neurospora crassa. Whereas in the wild-type strain...Full Text Available
Oryza sativa or Asian cultivated rice is one of the major cereal grass species domesticated for human food use during the Neolithic. Domestication of this species from the wild grass...Full Text Available
We have developed a new strategy for immunization of wild rabbit populations against myxomatosis and rabbit hemorrhagic disease (RHD) that uses recombinant viruses based on a naturally attenuated field...Full Text Available
Background and AimsMany plants reproduce both clonally and sexually, and the balance between the two modes of reproduction will vary among populations. Clonal reproduction was characterized...Full Text Available
Glandular trichomes of the wild tomato species Lycopersicon pennellii secrete 2,3,4-O-tri-acyl-glucose (-Glc), which contributes to insect resistance. A Glc acyltransferase...Full Text Available
BackgroundMany commercial banana varieties lack sources of resistance to pests and diseases, as a consequence of sterility and narrow genetic background. Fertile wild relatives,...Full Text Available
Low concentrations of sugars altered the sensitivity of seed germination to inhibition by exogenous abscisic acid (ABA). Germination of wild-type and ABA-insensitive (abi) Arabidopsis...Full Text Available
From diseased wild and cultured Oreochromis niloticus in Lower Egypt, 17 Aeromonas hydrophila isolates were recovered. The mortality was between 10% and 70% in among cultured fish. The course of the disease ran in an acute manner. For cultured fish, the disease outbreaks were found mainly in winter and for the wild Nile fish, mortalities were observed in late spring and summer. Additionally wild fish were affected with ectoparasites. The LD50 values of the isolates ranged between 10(3) and 10(7). Isolates of high virulence were resistant to 1 hr boiling and to the bactericidal effect of fresh normal guinea pig serum. Moreover, they did not agglutinate in acriflavin. Only the virulent isolates could agglutinate tilapia erythrocytes. The above effects were reversed for avirulent isolates while moderately virulent isolates showed no consistency in their reactions. Tube agglutination test using O and WC antisera prepared ...
The reaction of the retinylidene Schiff base in bacteriorhodopsin (bR) to the water-soluble reagent hydroxylamine is enhanced by greater than 2 orders of magnitude under illumination. We have used this reaction as a probe for changes in Schiff base reactivity during the photocycle of wild-type bR and mutants defective in proton transport. We report here that under illumination at pH 6, the D85N mutant has a 20-fold lower rate and the D212N mutant has a greater than 4-fold higher rate for the light-dependent reaction with hydroxylamine compared with wild-type bR. In contrast, the reactivities of wild-type bR and the D96N and T46V mutants are similar. It has been previously shown that the D96N and T46V replacements have no significant effect on the kinetics of M formation but have dramatic effects on rate of the decay of M. We therefore conclude that the hydroxylamine reaction occurs before formation of the M intermediate. ...
This report is intended to satisfy two concurrent needs: (1) provide a contract deliverable from Oncorh Consulting to the Washington Department of Fish and Wildlife (WDFW), with emphasis on identification of salient results of value to ongoing Yakima/Klickitat Fisheries Project (YKFP) planning and (2) summarize results of research that have broader scientific relevance. This is the fourth in a series of reports that address reproductive ecological research and monitoring of spring chinook populations in the Yakima River basin. This annual report summarizes data collected between April 1, 2004 and March 31, 2005 and includes analyses of historical baseline data, as well. Supplementation success in the Yakima Klickitat Fishery Project's (YKFP) spring chinook (Oncorhynchus tshawytscha) program is defined as increasing natural production and harvest opportunities, while keeping adverse ecological interactions and genetic impacts within acceptable bounds (Busack et al. 1997). ...
Wild and tissue cultured plants of Portulaca grandiflora Hook. have shown to be able to decolorize a sulfonated diazo dye Navy Blue HE2R (NBHE2R) up to 98% in 40h. A significant induction in the activities of lignin peroxidase, tyrosinase and DCIP reductase was observed in the roots during dye decolorization. The wild plants and tissue cultures could independently decolorize and degrade NBHE2R into metabolites viz. N-benzylacetamide and 6-diazenyl-4-hydroxynaphthalene-2-sulfonic acid. A dye mixture and a textile effluent were also decolorized efficiently by P. grandiflora. The phytotoxicity study revealed reduction in the toxicity due to metabolites formed after dye degradation.
We describe the cloning and characterization of a mutated thermostable DNA polymerase from Thermus aquaticus (Taq) that exhibits an increased reverse transcriptase activity and is therefore designated for one-step PCR pathogen detection using established real-time detection methods. We demonstrate that this Taq polymerase mutant (Taq M1) has similar PCR sensitivity and nuclease activity as the respective Taq wild-type DNA polymerase. In addition, and in marked contrast to the wild-type, Taq M1 exhibits a significantly increased reverse transcriptase activity especially at high temperatures (>60degreeC). RNA generally hosts highly stable secondary structure motifs, such as hairpins and G-quadruplexes, which complicate, or in the worst case obviate, reverse transcription (RT). Thus, RT at hi...
Summary 1. Plants express multiple defensive traits, but little is known about the genetic stability and phenotypic plasticity of these traits in nature. To investigate sources of variation and their potential ecological consequences for herbivores, we combined field observations of cyanogenic lima bean with laboratory experiments. 2. Field studies in South Mexico revealed a distinct variability of cyanogenic traits within and among wild lima bean populations. To differentiate among genetic variation and the impact of ambient conditions on plant phenotypes, we used seed grown plants as well as clones propagated from high (HC) and low cyanogenic (LC) wild type plants. 3. In growth chamber experiments, we cultivated plants under three intensities each of drought and salt stress, nutrient sup...
Abstract Crouzon syndrome is characterized by premature fusion of sutures and synchondroses. Recently, the first mouse model of the syndrome was generated, having the mutation Cys342Tyr in Fgfr2c, equivalent to the most common human Crouzon/Pfeiffer syndrome mutation. In this study, a set of micro-computed tomography (CT) scannings of the skulls of wild-type mice and Crouzon mice were analysed with respect to the dysmorphology caused by Crouzon syndrome. A computational craniofacial atlas was built automatically from the set of wild-type mouse micro-CT volumes using (1) affine and (2) non-rigid image registration. Subsequently, the atlas was deformed to match each subject from the two groups of mice. The accuracy of these registrations was measured by a comparison of manually placed landma...
A phytase gene (phyA), isolated from Aspergillus ficuum (AF537344), was introduced into cotton (Gossypium hirsutum L.) by Agrobacterium-mediated transformation to increase the phosphorus (P) acquisition efficiency of cotton. Southern and Northern blot analyses showed that the phyA was successfully incorporated into the cotton genome and expressed in transgenic lines. After growing for 45?days with phytate (Po) as the only P source, the shoot and root dry weights of the transgenic plants all increased by nearly 2.0-fold relative to those of wild-type plants, but were similar to those of transgenic plants supplied with inorganic phosphorus. The phytase activities of root extracts prepared from transgenic plants were 2.4- to 3.6-fold higher than those from wild-type plants, and the extracellu...
Despite efforts to control late blight in potatoes by introducing Rpi-genes from wild species into cultivated potato, there are still concerns regarding...Full Text Available
Previously it was shown that the host-range gene of the Bratislava strain of avain sarcoma virus (B77 virus) spontaneously mutates with a very high rate. The wild-type B77 virus called B77 virus-II,...Full Text Available
We report an analysis of four strains of baker's yeast (Saccharomyces cerevisiae) using biocavity laser spectroscopy. The four strains are grouped in two pairs (wild type and altered), in which one strain differs genetically at a single locus, affecting mitochondrial function. In one pair, the wild-type rho+ and a rho0 strain differ by complete removal of mitochondrial DNA (mtDNA). In the second pair, the wild-type rho+ and a rho- strain differ by knock-out of the nuclear gene encoding Cox4, an essential subunit of cytochrome c oxidase. The biocavity laser is used to measure the biophysical optic parameter Deltalambda, a laser wavelength shift relating to the optical density of cell or mitochondria that uniquely reflects its size and biomolecular composition. As such, Deltalambda is a powerful parameter that rapidly interrogates the biomolecular state of single cells and mitochondria. Wild-type ...
DescriptionThis project is part of the BBSRCs special initiative on plant and microbial metabolomics. The project will primarily focus on the trichothecene mycotoxin producing Ascomycete fungus Fusarium graminearum (Fg) which causes ear blight disease of small grain cereals. The project aims to explore the metabolome of various wild-type and single gene deletion Fg strains and to compare some of these with the identical gene mutation in the budding yeast, S. cerevisiae (Sc) and the saprophytic filamentous [continued...
... The larvae's only hope for survival is if the antibiotic tetracycline is present -because it mops up the enzyme. The developers hope male GM mosquitoes will compete with normal males for females so that repeated releases cut numbers of A. aegypti in dengue-prone areas. The mosquitoes will be released in the inland districts of Bentong in the state of Pahang, and Alor Gajah and Melaka in the state of Malacca, according to the National Biosafety Board. Each location will have two release phases: the first at a site ...
Recombinant plasmids containing mutant or wild-type adenovirus serotype 2 EIa genes that produce the 12S mRNA alone, the 13S mRNA alone, or both mRNAs were cotransfected into HeLa cells with plasmids...Full Text Available
Biglycan (bgn)-deficient mice (KO) have defective osteoblasts which lead to changes in the amount and quality of bone. Altered tissue strength in C57BL6/129 (B6;129) KO mice, a property which...Full Text Available
... The male GM mosquitoes mate with normal females to produce larvae that die unless the antibiotic tetracycline is present. In tetracycline's absence an enzyme accumulates to a toxic level, killing the larvae. The developers hope the strategy could be combined with other mosquito control methods to reduce transmission in dengue-prone areas. Ricarda Steinbrecher, a geneticist and co-director of EcoNexus -a UK-based non-profit research organisation -expressed surprise ...
... The male GM mosquitoes mate with normal females to produce larvae that die unless the antibiotic tetracycline is present. In tetracycline's absence an enzyme accumulates to a toxic level, killing the larvae. The developers hope the strategy could be combined with other mosquito control methods to reduce transmission in dengue-prone areas. Ricarda Steinbrecher, a geneticist and co-director of EcoNexus - a UK-based non-profit research organization - ...
Rice, the world's major staple crop, is a poor source of essential micronutrients, including folates (vitamin B9). We report folate biofortification of rice seeds achieved by overexpressing two Arabidopsis thaliana genes of the pterin and para-aminobenzoate branches of the folate biosynthetic pathway from a single locus. We obtained a maximal enhancement as high as 100 times above wild type, with 100 g of polished raw grains containing up to four times the adult daily folate requirement.
We report that expression of a nearly full-length cDNA clone of the L-A double-stranded RNA virus causes virus loss in a wild-type strain of Saccharomyces cerevisiae. We show that in this system exclusion...Full Text Available
In the paper, a potential of solar energy for drying of medicinal plants in Polish conditions is estimated and development of solar drying technologies is presented. The results of economic assessment of flat-plate solar collectors applied for drying of medicinal plants on a farm are promising. In some specific conditions, e.g. drying of wild grown medicinal plants in remote areas, even application of photovoltaic modules for driving of a fan of a solar dryer is a profitable option and enables easy control of the drying air temperature.
Little is known about how diet and energy metabolism interact in determination of lifespan under ad libitum feeding. From 12 weeks of age until death, male and female wild-type (WT) and transgenic (TG)...Full Text Available
The nr1 soybean (Glycine max [L.] Merr.) mutant does not contain the two constitutive nitrate reductases, one of which is responsible for enzymic conversion of nitrite to...Full Text Available
Yeast arginyl-tRNA synthetase recognizes the non-modified wild-type transcripts derived from both yeast tRNA(Arg) and tRNA(Asp) with equal efficiency. It discriminates its cognate natural substrate,...Full Text Available
Eribulin is a new anticancer agent currently in Phase III clinical trials for the treatment of metastatic breast cancer. In the current studies, we have investigated the effects of P-glycoprotein (P-gp) on the in vivo disposition of eribulin using CF-1 abcb1a-deficient mice, and the influence of eribulin on P-gp-mediated efflux of digoxin in Caco-2 cells. Eribulin was administered intravenously and orally in both CF-1 wild-type and CF-1 abcb1a-deficient mice. P-gp-mediated efflux of digoxin in Caco-2 cell monolayers was measured in the presence of eribulin. The plasma exposure to eribulin was higher in CF-1 abcb1a-deficient mice than that in CF-1 wild-type mice after intravenous (IV) and oral (PO) administrations. The oral bioavailability of eribulin was 62.3% in CF-1 abcb1a-deficient mice...
Experiments were performed to investigate the involvement of the cell membrane in the excision DNA repair process in Escherichia coli. Two membrane-binding drugs, procaine and phenethyl alcohol (PEA), inhibited liquid-holding recovery (LBR) in u.v.-irradiated E. coli wild-type and recA strains. In uvrB and polA strains where, after u.v.-irradiation, LHR was absent the two drugs had no effect. Both drugs markedly reduced the removal of u.v.-induced thymine dimers in the DNA of wild-type cells (H/r30). Analysis by alkaline sucrose gradients revealed that PEA inhibited the incision step in excision repair. In contrast, procaine had no effect on incision but apparently inhibited the late steps in excision repair. PEA dissociated DNA from the cell membrane, whereas procaine did not. The results suggest that the two drugs PEA and procaine inhibit LHR and the excision repair process operating on u.v.-induced damage in E. coli by at least two different ...
The resistance of six tropical hardwoods to fungal degradation by two wild-type strains of Phanerochaete chrysosporium Burdsall was investigated using vermiculite burial and wood-block weight loss techniques. Radiation-induced wood-polymer composites (WPC), based on two hardwoods Ramin and Rubberwood with methyl methacrylate, were prepared, and samples were also exposed to the wood-rotting fungus. A significant improvement in resistance to fungal decay was observed in the WPC. Scanning-electron micrographs of the two woods and their composites after fungal degradation are presented and discussed.
DescriptionBoth wild and aquaculture reared abalone, mussels and other shellfish of commercial importance as a food resource to man can be afflicted by poisoning if they feed on toxic species of small single-celled planktonic marine algae. If humans consume these shellfish that have become intoxicated by HAB's, illness or even death may occur. HAB's occur world-wide, but some regions are more severely affected than others. The incidence of HAB's also appear to be on the increase globally, perhaps because o [continued...
This work has carried out a molecular structure characterization of two Brazilian plants, Harpalyce brasiliana Benth and Bredemeyera floribunda Wild, using {sup 1} H and {sup 13} C NMR spectroscopy. NMR spectra were presented and analysed, then several structures have been proposed based on spectral data, and finally the structural determination was done 3 refs., 4 figs., 1 tab.
The transfer of fish from field to laboratory facilities or their propagation in closed or restricted systems frequently results in bacterial infection and ultimately large-scale mortality. In attemps to alleviate this problem, we have added tetracycline hydrochloride to the water prophylactically (pretreating tanks before wild fish were added) and therapeutically (treating tanks after bacterial outbreaks were detected.) In the present study, we examined the effect of tetracyline hydrochloride on the critical thermal maximum (CTM) of the common shiner (Notropis cornutus).
Some of the known toxic effects of lead in mammals including man are, impaired heme synthesis, anemia, nepatopathy, nephropathy, behavioral disorders and neuropathy. However, very little is known about the effect of lead on endocrine physiology. Some data are available on lead induced impairment of thyroid function in occupationally exposed men and experimental rats. As lead nitrate is largely consumed through water and food, in this study the wild rodents Funambulus pennanti were administered lead through their drinking water and their thyroid structure, radioiodine 131-I percentage uptake and protein bound iodine (PBI) level were assessed.
The authors have developed a complex of activation methods of analysis using a nuclear reactor (nuclear activation analysis) and a cyclotron (charged-particle activation analysis). The methods have been used to determine the concentrations of more than 20 elements in five medicinal plants native to Uzbekistan: Syrian rue (Peganum harmala L.), plantain (Plantago lanceolata), peppermint (Mentha piperata L.), sage (Salvia officinalis L.), and ziziphora (Ziziphora bungeana Yur.). The results of radio-activation analysis were compared with the results of standard spectral analysis performed in another laboratory and the accuracy of the procedures developed was evaluated on the basis of the results.
Abstract The white sea bream, Diplodus sargus (Teleostei, Sparidae), is a species with a high commercial importance in Mediterranean aquaculture. There is currently little information available about the genetic characteristics of cultured populations. In this survey, we have developed eight polymorphic microsatellites for the white sea bream using an enriched genome library protocol. All of them were polymorphic in the 67 individuals tested, 32 of which were wild specimens, and 35 were individuals from a captive F1 broodstock. These markers can potentially be useful tools for use in population genetic studies.
Chorismate mutase, a branch-point enzyme in the aromatic amino acid pathway of Saccharomyces cerevisiae, and also a mutant chorismate mutase with a single amino acid substitution in the C-terminal part of the protein have been purified approximately 20-fold and 64-fold from overproducing strains, respectively. The wild-type enzyme is activated by tryptophan and subject to feedback inhibition by tyrosine, whereas the mutant enzyme does not respond to activation by tryptophan nor inhibition by tyrosine. Both enzymes are dimers consisting of two identical subunits of M_r 30,000, each one capable of binding one substrate and one activator molecule. Each subunit of the wild-type enzyme also binds one inhibitor molecule, whereas the mutant enzyme lost this ability. The enzyme reaction was observed by "1H NMR and shows a direct and irreversible conversion of chorismate to prephenate without the accumulation of any enzyme-free intermediates. The ...
Reproductive status influences cell proliferation and the survival of new cells in the dentate gyrus of adult laboratory-reared and wild female meadow voles; reproductively inactive (RI) females have more proliferating cells and more labeled cells that survive 5 weeks vs. 2 h than reproductively active (RA) females. However, the effect of season has only been studied in a wild sample of male meadow voles and factors such as age and experience that have been shown to influence neurogenesis in the dentate gyrus of adult mammals cannot be controlled in a wild sample. Therefore, we investigated whether reproductive status regulates neurogenesis (cell proliferation and/or the survival of new cells) in the dentate gyrus of laboratory-reared adult male meadow voles so that confounding variables could be controlled. Males were acclimated to a short- or a long-photoperiod to simulate the non-breeding or breeding season, ...
The sterile insect release technique can often be improved by removal of females before release. Rearing efficiencies can also be increased if removal of the females occurs at early developmental stages. In order to begin to develop genetic sexing strains for the pink bollworm, Pectinophora gossypiella (Saunders), it was necessary to determine the best dosage of radiation for induction of viable sex linked recessive lethal mutations and to see if stocks containing induced sex linked lethals could be maintained in culture. Sex linked recessive lethal mutations can be detected by distorted sex ratios in the progeny of treated adults. However, in the pink bollworm, highly distorted sex ratios are common even in the absence of induced mutations. Therefore, a visible sex linked trait, purple eye, was used as a marker for the untreated X-chromosomes in crosses. Thus, the presence of a recessive sex linked lethal mutation induced by the radiation treatment may be detected by the combination ...
A single amino acid substitution (Asp #-># Asn) at position 138 of E. coli EF-Tu was induced in the tufA gene by an M13 phage oligonucleotide site-directed mutagenesis protocol. The mutated tufA gene was then subcloned in a plasmid vector and expressed in maxicells. The properties of ["3"5S]methionine labelled mutant and wild type EF-Tu's were compared by in vitro assays. Mutant and wild-type EF-Tu's bound EF-Ts with approximately equal affinities. The 138-Asn mutation greatly reduced the protein's affinity for GDP; however, this mutation dramatically increased the proteins affinity for XDP. The mutant protein forms a stable complex with phe-tRNA and XTP, which binds to ribosomes; whereas, it does not form a complex with phe-tRNA and GTP. These results suggest that in EF-Tu x NDP complexes amino acid residue 138 must interact with the substituent on C-2 of the purine ring. Thus in wild-type EF-Tu Asp-138 would H-bond to ...
A new area-wide pest control strategy using the pink bollworm, Pectinophora gossypiella (Saunders) (Lepidoptera: Gelechiidae), genetically transformed with a conditionally lethal gene, is under development. Conditional lethality of several transgenic pink bollworm strains was demonstrated in a series of laboratory rearing experiments. Pink bollworms were transformed with genetic constructs using the RIDL technology (Release of Insects with a Dominant Lethal gene) for development of an autocidal biological control system for possible supplement or replacement of radiation based sterile insect release. LA1124 is a lethal construct controlled by a tetracycline repressible transactivator protein (tTA), in which binding of tTA to its specific target sequence tetO drives production of more tTA. In the absence of tetracycline, this leads to lethality by high expression of tTA. When tetracycline is present, tTA does not bind tetO, and so the positive feedback cycle is not established and tTA ...
Neuron differentiation is a complex process involving various cell-cell interactions, and multiple signaling pathways. We showed previously that CD40 is expressed and functional on mouse and human neurons. In neurons, ligation of CD40 protects against serum withdrawal-induced injury and plays a role in survival and differentiation. CD40 deficient mice display neuron dysfunction, aberrant neuron morphologic changes, and associated gross brain abnormalities. Previous studies by Tone and colleagues suggested that five isoforms of CD40 exist with two predominant isoforms expressed in humans: signal-transducible CD40 type I and a C-terminal truncated, non-signal-transducible CD40 type II. We hypothesized that differential expression of CD40 isoform type I and type II in neurons may modulate neuron differentiation. Results show that adult wild-type, and CD40"-"/"- deficient mice predominantly express CD40 type I and II isoforms. Whereas adult ...
Summary The objective of this study was to examine the role of root carbohydrate levels and metabolism in the waterlogging tolerance of contrasting mung bean genotypes. An experiment was conducted with two cultivated mung bean (Vigna radiata) genotypes viz., T44 (tolerant) and Pusa Baisakhi (PB) (susceptible), and a wild Vigna species Vigna luteola under pot-culture to study the physiological and molecular mechanism of waterlogging tolerance. Waterlogging resulted in decrease in relative water content (RWC), membrane stability index (MSI) in root and leaf tissues, and chlorophyll (Chl) content in leaves, while the Chl a/b ratio increased. Waterlogging-induced decline in RWC, MSI, Chl and increase in Chl a/b ratio was greater in PB than V. luteola and T44. Waterlogging caused decline in tot...
The Kemp's Ridley sea turtle, Lepidochelys kempi, is an endangered species. The National Marine Fisheries Service's Head Start program is part of an international operation to save the turtles from extinction. Under the Head Start program, eggs from the Ridley's only known wild nesting beach at Rancho Nuevo in Mexico are transported to Padre Island on the Texas coast to be hatched. The head start enables the turtles to develop a survival advantage. The principal objective was to develop baseline waste-characterization data required to design a waste-water treatment scheme for the Galveston Head Start facility. As a secondary objective, preliminary testing of some filtration components was undertaken to determine which units were most appropriate for inclusion in a wastewater treatment scheme.
The effects of eight chemicals on the inactivation rate of ultraviolet radiation on the colony building capabilities of two strains of Saccharomyces cervisae - a wild type strain and a mutant deficient in excision repair - were studied. The insecticide methoxychlor, the herbicide 2,4-dichlorophenoxyacetic acid, the fungicide pentachlorophenol and its metabolite tetrachlorohydroquinone, as well as the chemicals acrylonitrile and 2,3-dichloro-1-propene have no significant impact on the effects of UV radiation in Saccharomyces cerevisae. Depending on the concentration, trichloroethylene increases the sensitivity to UV radiation. The herbicide paraquat provides efficient protection against UV radiation at concentrations where a toxic effect cannot be observed even without UV. The results were rather similar for both strains. (orig.).
Abstract Intrasexual selection can occur through direct aggressive interactions between males for access to females. We tested the relationship between social dominance and male reproductive success among meadow voles, Microtus pennsylvanicus. Dominance ranks of wild-caught males were determined using neutral arena trials, with the winner of two of three trials considered dominant. These males were then released into field enclosures and allowed to visit females housed in nestboxes for 8 wk, and males' home range sizes were determined using weekly grid trapping. Male reproductive success was determined using molecular paternity analysis (six microsatellite primers) for all pups born during the field experiment. Males with higher dominance ranks had larger home ranges. However, male dominan...
In order to investigate the potential role of arctic geese in the epidemiology, the spatial and temporal spread of selected avian diseases, in autumn 2002, a virological and serological survey designed as capture-mark-resighting study was conducted in one of the most important coastal resting sites for migratory waterfowl in Germany. Oropharyngeal, cloacal swabs and blood samples were collected from a total of 147 birds comprising of three different arctic geese species including White-fronted Goose (Anser albifrons), Tundra Bean Goose (Anser fabalis rossicus), Pink-footed Goose (Anser brachyrhynchus) as well as from 29 non-migratory Canada Geese (Branta canadensis). Altogether, six adeno-like viruses (ALV; 95% CI, 1.74?9.92%) and two avian paramyxoviruses (APMV-4; 95% CI, 0.19?5.53%) were...
We quantified the resistance levels of transgenic rice plants, expressing Myxococcus xanthus protoporphyrinogen oxidase (PROTOX) in chloroplasts and mitochondria, to PROTOX inhibitors, acifluorfen, oxyfluorfen, carfentrazone-ethyl, and oxadiazon. We also determined whether active oxygen species-scavenging enzymes are involved in the resistance mechanism of transgenic rice. The transgenic rice line M4 was about >200-fold more resistant to oxyfluorfen than the wild-type (WT). M4 was also resistant to acifluorfen, carfentrazone-ethyl, and oxadiazon, but did not show multiple resistance to imazapyr and paraquat, which have different target sites. Acifluorfen, oxyfluorfen, carfentrazone-ethyl, and oxadiazon reduced the chlorophyll content in leaves of WT, but had minimal or no effect on M4. The...
Using a novel, pulsed micro-second time-resolved photoacoustic (PA) instrument, we measured thermal dissipation and energy storage (ES) in the intact cells of wild type (WT) Chlamydomonas reinhardtii, and mutants lacking either PSI or PSII reaction centers (RCs). On this time scale, the kinetic contributions of the thermal expansion component due to heat dissipation of absorbed energy and the negative volume change due to electrostriction induced by charge separation in each of the photosystems could be readily distinguished. Kinetic analysis revealed that PSI and PSII RCs exhibit strikingly different PA signals where PSI is characterized by a strong electrostriction signal and a weak thermal expansion component while PSII has a small electrostriction component and large thermal expansion....
Recycling of carbon by the photorespiratory pathway involves enzymatic steps in the chloroplast, mitochondria, and peroxisomes. Most of these reactions are essential for plants growing under ambient CO2 concentrations. However, some disruptions of photorespiratory metabolism cause subtle phenotypes in plants grown in air. For example, Arabidopsis thaliana lacking both of the peroxisomal malate dehydrogenase genes (pmdh1pmdh2) or hydroxypyruvate reductase (hpr1) are viable in air and have rates of photosynthesis only slightly lower than wild-type plants. To investigate how disruption of the peroxisomal reduction of hydroxypyruvate to glycerate influences photorespiratory carbon metabolism we analyzed leaf gas exchange in A. thaliana plants lacking peroxisomal HPR1 expression. In addition, b...
We describe the analysis of forward mutations induced in the tetracycline resistance gene of the plasmid pBR322 by directing the reaction of the carcinogen N-acetoxy-N-2-acetylaminofluorene (N-AcO-AAF) to a small restriction fragment (BamHI, SalI) that is located in the proximal part of the antibiotic-resistance gene. Mutant plasmids obtained both in wild type and excision repair deficient (uvrA) bacterial cells are compared. Preliminary data showing the distribution of the -AAF adducts along this restriction fragments are discussed in relation to the observed spectrum of mutations. 20 references, 4 figures.
In Europe, the use of anti-EGFR monoclonal antibodies is restricted to Kirsten RAS (KRAS) wild-type colorectal tumors. Information on the KRAS status of the patients tumor is thus key for clinical practice; however, there is little guidance or definition on which KRAS mutations to assess and how to assess them. To ensure the consistency and the quality of KRAS test results in Europe, an interlaboratory control network needs to be set up. This pilot study aimed to identify the variables that need to be assessed in a quality control scheme and to provide a first assessment in a selected set of laboratories. Fourteen different tumor cases were circulated between 13 laboratories by a central laboratory acting as the referent for the mutation status determination. This study illustrated that of...
Laboratory studies have been carried out to determine the toxicity of methiocarb pellets to wild trapped wood mice in order to provide some background data prior to any further evaluation of hazard in the field. In this study, wood mice were exposed to dry and to dampened methiocarb pellets in order to reproduce field trial application conditions. Field observations of methiocarb pellets indicate that the physical character changes under dry and wet weather conditions. This may affect their relative attractiveness and potential toxicity to wood mice. The laboratory assessment of exposed wood mice included measurement of brain esterase activities, methiocarb residues in selected mouse tissue, carcasses, and histological evaluation of kidney, liver and lungs.
Numerous studies have shown that large, herbivorous waterfowl can reduce quantity of aquatic plants during the breeding or wintering season, but relatively few document herbivory effects at staging areas. This study was done to determine if feeding activities of tundra swans (Cygnus columbianus columbianus) and Canada geese (Branta canadensis) had a measurable additive influence on the amount of aquatic plants, primarily muskgrass (Chara vulgaris), wild celery (Vallisneria americana), and sago pondweed (Potamogeton pectinatus), removed during the fall migration period at Long Point, Lake Erie, Ontario. Exclosure experiments done in fall 1998 and 1999 showed that, as compared to ducks and abiotic factors, these two large herbivorous waterfowl did not have any additional impact on above or b...
One of the research challenges in oncology is to develop new biochemical methods for noninvasive tumor therapy evaluation to determine whether the chemotherapeutics is effective. Vascular endothelial growth factor (VEGF) was labeled with radioiodine and evaluated in vitro as well as in vivo, using A2058, a melanoma cell line overexpressing VEGFR-1 and -2. Saturation binding analysis with [{sup 125}I]-VEGF resulted in a K {sub d} of 0.1 nM. Internalization assays indicate the preserved ligand induced internalization and metabolization of the tracer. Biodistribution studies with [{sup 123}I]-VEGF in wild type and A2058 tumor-bearing athymic mice showed low background activity and a tumor to reference tissue ratio of maximum 6.12. These results suggest that [{sup 123}I]-VEGF is a potentially suitable tracer for tumor therapy evaluation.
Mutations in sclerostin function or expression cause sclerosing bone dysplasias, involving decreased antagonism of Wnt/Lrp5 signaling. Conversely, deletion of the VHL tumor suppressor in osteoblasts, which stabilize HIF-a isoforms and thereby enables HIF-a/b-driven gene transcription, increases bone mineral content and cross-sectional area compared to wild-type controls. We examined the influence of cellular hypoxia (1% oxygen) upon sclerostin expression and canonical Wnt signaling. Osteoblasts and osteocytes cultured under hypoxia revealed decreased sclerostin transcript and protein, and increased expression and nuclear localization of activated b-catenin. Similarly, both hypoxia and the hypoxia mimetic DFO increased b-catenin gene reporter activity. Hypoxia and its mimetics increased exp...
We investigated gathering and cultivating practices and how they complement each other in a rural population of Northwestern Patagonia. We analyzed plant diversity, species similarity, biogeographic origin, and plant use by means of semi-structured interviews and field visits. Pichi Leufu inhabitants used 173 species: 138 cultivated plants, mainly for edible purposes, and 45 wild species principally for medicinal use. Most cultivated species were exotic (91.3%), whereas gathered plants were both native and exotic. While locals maintained vegetable gardens, the adoption of greenhouses improved conditions for certain crops. The integration of novel practices with ancestral knowledge suggests resilient processes in this community, probably reflected in the dynamics of current horticultural an...
The function of selenium in an organism is mediated mostly by selenoproteins including glutathione peroxidase. Glutathione peroxidase is a potent anti-oxidative enzyme, scavenging a variety of peroxides. The green alga Scenedesmus quadricauda was used to investigate the relationship between the toxicity of selenium and the glutathione peroxidase activity. Selenium resistant strains SeIV and SeVI were synchronized and grown in high concentrations of Se (selenite or selenate). As a measure of selenium toxicity the EC50 values were determined. During growth of the untreated wild type, glutathione peroxidase activity increased slightly and then declined gradually until the end of the cell cycle. A similar pattern was observed in untreated resistant strains and when resistant strains were grown...
The integral membrane protein synaptophysin is one of the most abundant polypeptide components of synaptic vesicles. It is not essential for neurotransmission despite its abundance but is believed to modulate the efficiency of the synaptic vesicle cycle. Detailed behavioral analyses were therefore performed on synaptophysin knockout mice to test whether synaptophysin affects higher brain functions. We find that these animals are more exploratory than their wild type counterparts examining novel objects more closely and intensely in an enriched open field arena. We also detect impairments in learning and memory, most notably reduced object novelty recognition and reduced spatial learning. These deficits are unlikely caused by impaired vision, since all electroretinographic parameters measur...
The full length cDNA of the Brn1 was first cloned, and then expression of the Brn1 was analyzed and the function was identified by silencing technology. Results show that the full length cDNA of the C. lunata Brn1 gene contains 1001 base pairs and an 801?bp open reading frame encoding 267 amino acids. Semi-quantitative PCR analysis shows that the expression of Brn1 at 96?h is significantly higher than at 24 and 72?h (p?Brn1-silenced transformants were light brown in culture filtrate, and have significantly reduced toxin production relative to the wild-type. These results imply that Brn1 gene in C. lunata is not only involved in 1,8-dihydroxynaphthalene melanin synthesis, but is also relatively associated with toxin biosynthesis of the pathogen.
Mutations or multiplications in a-synuclein gene cause familial forms of Parkinson disease or dementia with Lewy bodies (LB), and the deposition of wild-type a-synuclein as LB occurs as a hallmark lesion of these disorders, collectively referred to as synucleinopathies, implicating a-synuclein in the pathogenesis of synucleinopathy. To identify modifier genes of a-synuclein-induced neurotoxicity, we conducted an RNAi screen in transgenic C. elegans (Tg worms) that overexpress human a-synuclein in a pan-neuronal manner. To enhance the RNAi effect in neurons, we crossed a-synuclein Tg worms with an RNAi-enhanced mutant eri-1 strain. We tested RNAi of 1673 genes related to nervous system or synaptic functions, and identified 10 genes that, upon knockdown, caused severe growth/motor abnormalit...
We describe the cloning and characterization of a mutated thermostable DNA polymerase from Thermus aquaticus (Taq) that exhibits an increased reverse transcriptase activity and is therefore designated for one-step PCR pathogen detection using established real-time detection methods. We demonstrate that this Taq polymerase mutant (Taq M1) has similar PCR sensitivity and nuclease activity as the respective Taq wild-type DNA polymerase. In addition, and in marked contrast to the wild-type, Taq M1 exhibits a significantly increased reverse transcriptase activity especially at high temperatures (>60 degrees C). RNA generally hosts highly stable secondary structure motifs, such as hairpins and G-quadruplexes, which complicate, or in the worst case obviate, reverse transcription (RT). Thus, RT at high temperatures is desired to weaken or melt secondary structure motifs. To demonstrate the ability of Taq M1 for RNA detection of pathogens, we ...
Genes encoding mutants of the thymine photodimer repair enzyme from bacteriophage T4 (T4 endonuclease V) having an amino acid substitution (T127M, W128A, W128S, Y129A, K130L, Y131A, Y132A) were constructed by use of a previously obtained synthetic gene and expressed in Escherichia coli under the control of the E. coli tryptophan promoter. An in vitro assay of partially fractionated mutant proteins for glycosylase activity was performed with chemically synthesized substrates containing a thymine photodimer. T127M and K130L showed almost the same activity as the wild-type protein. Although W128S, Y131A, and Y132A were slightly active, W128A and Y129A lost activity. The results indicated that the aromatic amino acids around position 130 may be important for the glycosylase activity. Mutant T127M was purified, and the Km value was found to be of the same order as that of the wild type (10(-8) M). In vivo activities for all mutants were ...
Sufficient genetic diversity exists in Indian tree species to give large genetic gains. Improvement potential, using wild gene resources for selection and breeding is vast as there are over 244 genera and 400 forest tree species in India. Of these over 30 species and 22 genera are of outstanding forestry value. Selective genetic-variation can be lifted from wild populations, multiplied vegetatively or by seed, and planted with controlled germplasm sources. Among the Indian species no less than 74 reproduce by cutting, 11 by layers, 9 by grafting and buding and 104 by root suckers. More than 161 species coppice vigorously. Afforestation techniques of over 170 species have been worked out. Tree genetic approach of survey of natural variability, quick evaluation, selection, conservation multiplication and planting superior variability directly in the field can give immediate genetic gains. This article discusses genetic information on indigenous ...
Perinatal exposure to cadmium (Cd) or methylmercury (MeHg) results in impaired neurodevelopment. Thyroid hormone is essential for normal brain development. However, the issue whether Cd or MeHg, especially at low doses, interrupts thyroid hormone action remains to be investigated. In the present study, effects of perinatal exposure to low levels of Cd or MeHg on thyroid hormone metabolism were examined using metallothionein I and II (MT-I/II) null or wild-type neonatal mice. Dams were exposed to 10 mg/L water of Cd or 5 mg/kg chow of MeHg from gestational day 0 to post-natal day 10 (PND 10). Sera, livers and brains were collected from neonates on PND 10. Iodothyronine deiodinase activities and serum thyroxine (T4) concentrations were measured. MeHg exposure failed to induce changes in serum T4 levels and liver type 1 deiodinase (D1) and brain type 2 deiodinase (D2) activities regardless of the MT genotype. However, exposure to MeHg resulted in a decrease in brain ...
A 450 MeV Synchrotron Radiation Source, Indus-1 is being constructed at Centre for Advanced Technology at Indore in central India. This paper narrates our experience in construction and alignment of synchrotron machines which was first of its kind for most of us. Careful design, planning and execution of the work yielded modest results so that alignment accuracies between 0.1 to 0.3 mm could be achieved (in booster synchrotron) and, which have proved to be adequate up till now. The building of the SRS complex was constructed such that machine rings have their foundations isolated from rest of the building. A number of 100 x 150 mm size steel plates were embedded in the concrete of floor and walls to serve as base for reference surveying marks which were established later. The pre-injector and booster ring are enclosed in a radiation shielding zone with separate ventilation system. Dipoles, quadrupoles and a few sextupoles were fiducialised during their field mappings; on a CNC ...
Apoptosis, or programmed cell death (PCD), is a fundamental process that protects organismal integrity. In earlier work, we demonstrated that over-expression of either of two anti-apoptotic members of the BCL-2 family (BCL-2 or BCL-X L could elevate the frequency of radiation-induced mutations at the autosomal TK1 locus in human TK6 lymphoblasts that express wild-type TP53. Ectopic expression of BCL-X L also elevated the frequencies of double-strand break-induced gene conversion. The purpose of this study is to determine if BCL-2 family proteins promote radiation mutagenesis indirectly through their suppression of PCD, or whether the 'pro-mutagenic' function of these proteins can be separated from their anti-apoptotic function. We developed stable transfectants of TK6 cells that express a mutated form of BCL-X L with a single amino acid substitution in the BH1 domain that is known to interfere with the ability to suppress PCD (BCL-X L gly159ala). We also developed ...
Abstract Bartonella are arthropod-borne, fastidious, Gram-negative, and aerobic bacilli distributed by fleas, lice, sand flies, and, possibly, ticks. The zoonotic Bartonella species, Bartonella henselae and Bartonella clarridgeiae, which are the causes of cat scratch disease and endocarditis in humans, have been reported from cats, cat fleas, and humans in Australia. However, to date, there has been no report of B. henselae or B. clarridgeiae in Australian wild animals and their ectoparasites. B. henselae and B. clarridgeiae were detected in fleas (Ctenocephalides felis) from red foxes (Vulpes vulpes), an introduced pest animal species in Australia, and only B. clarridgeiae was detected in blood from one red fox. Phylogenetic analysis of the ribosomal intergenic spacer region revealed that the B. henselae detected in the current study were related to B. henselae strain Houston-1, a major pathogenic strain in humans in Australia, and confirmed the genetic ...
The trace element selenium (Se) is required in the production of enzymes that protect mammalian cells from oxidative damage due to the byproducts of aerobic respiration. Its deficiency in livestock results in the nutritional muscular dystrophy called white muscle disease. This especially affects juveniles in the preweaning period. Symptoms have also been reported in wild herbivores on low-Se forage, and their appearance may be episodic, suggesting temporal variations in Se uptake by plants. Here, we report variations in selenium concentrations in Carex spp. sedges used as forage by bighorn sheep (Ovis canadensis) on summer range in the Wyoming, Wind River Mountains, and correlate those variations with soil conditions that affect the bioavailability of selenium. Variations in available Se are explained based upon the known oxidation/reduction chemistry of the element. It is concluded that water-saturated, alpine soils provide conditions suitable for the reduction of ...
The authors report that the replacement of Leu-93 in bacteriorhodopsin by Ala (L93A) or Thr (L93T) slows down the photocycle by approximately 100-fold relative to wild-type bacteriorhodopsin. Time-resolved visible absorption spectroscopy and resonance Raman experiments, respectively, show the presence of long-lived O-like and N-like intermediates in the photocycles of the above mutants. We infer the existence of an equilibrium between the N and O intermediates in the photocycles of these mutants. The L93A and L93T mutants exhibit normal proton pumping under continuous illumination, suggesting that the decay of the N and/or O intermediate, and consequently, proton translocation, can be accelerated by the absorption of a second photon. Since the 13-cis----all-trans reisomerization of retinal is completed during the decay of the N and O intermediates, they conclude that the interaction of Leu-93 with retinal is important in this phase of the photocycle. This ...
The nuclear pore complexes (NPCs) reversibly disassemble and reassemble during mitosis. Disassembly of the NPC is accompanied by phosphorylation of many nucleoporins although the function of this is not clear. It was previously shown that in the transmembrane nucleoporin gp210 a single serine residue at position 1880 is specifically phosphorylated during mitosis. Using amino acid substitution combined with live cell imaging, time-lapse microscopy and FRAP, we investigated the role of serine 1880 in binding of gp210 to the NPC in vivo. An alanine substitution mutant (S1880A) was significantly more dynamic at the NPC compared to the wild-type protein, suggesting that serine 1880 is important for binding of gp210 to the NPC. Moreover a glutamate substitution (S1880E) closely mimicking phosphorylated serine specifically interfered with incorporation of gp210 into the NPC and compromised its post-mitotic recruitment to the nuclear envelope of daughter nuclei. Our ...
Cholera vaccine candidate Peru-15 was derived from a Vibrio cholerae O1 El Tor Inaba strain by deleting the cholera toxin genetic element, introducing the gene encoding cholera toxin B subunit into recA, and screening for nonmotility. In a controlled study, Peru-15 (2 x 10(8) cfu) was administered to 11 volunteers. No vaccinee developed diarrhea, and 10 of 11 had > 4-fold rises in vibriocidal antibody titers. One month later, 5 vaccinees and 5 control volunteers were challenged with wild type V. cholerae O1. Four of 5 controls developed diarrhea (mean, 1.9 L). Two Peru-15 vaccinees developed diarrhea, 1 with < 0.3 L and 1 with approximately 1.0 L; this latter volunteer had not developed a significant vibriocidal immune response to vaccination. Peru-15 shows promise as a single-dose, oral cholera vaccine that is safe, immunogenic, and protective. PMID:7561195
Dehydrins, a subfamily of group 2 LEA proteins, are intrinsically unstructured plant proteins that accumulate in the late stages of seed development and in vegetative tissues subjected to water deficit, salinity, low temperature, or abscisic acid treatment. In this study, we isolated and characterized ZmDHN2b, a maize dehydrin gene. The genomic organization of the ZmDHN2b gene and its expression in maize seedlings were analyzed. To investigate the function of ZmDHN2b, we generated transgenic tobacco plants constitutively overexpressing ZmDHN2b. Ectopic expression of ZmDHN2b in tobacco accelerated seed germination and seedling growth at 15?C. Furthermore, ZmDHN2b-overexpressing lines had lower levels of cold-induced malondialdehyde and less electrolyte leakage than wild-type tobacco at 4?C....
An overview is given of several computer models in use in the Netherlands to determine and analyze the quality of air and the effect of air pollution abatement measures. [Dutch] Voor het vaststellen van de luchtkwaliteit of het bepalen van het effect van emissiereducerende maatregelen op die luchtkwaliteit zijn in ons land meerdere computermodellen beschikbaar. In het kader van het Vooronderzoek Innovatieprogramma Luchtkwaliteit (VIPL) wilde het ministerie van Verkeer en Waterstaat meer inzicht in de betrouwbaarheid en specifieke eigenschappen van deze modellen. Deze studie geeft daarom een overzicht en analyse van de verschillen, overeenkomsten en voor- en nadelen van de meest gangbare Nederlandse computermodellen om de luchtkwaliteit rond snelwegen te bepalen. Dit zijn: Het Voorspellingssysteem Luchtkwaliteit Wegtraces (VLW); CAR-II; Het TNO-verkeersmodel; Het Heaven-model van TNO; Urbis (air); en NNM+ (KEMA-STACKS)
In vitro mutagenesis of functional DNA gene fragments by covalently reactive agents permits one in principle to examine the consequent alterations in DNA sequence directly. I have carried out selective mutagenesis of the tetracycline resistance gene in the plasmid pBR322 using the long wavelength UV light activated reaction of 4,5',8-trimethylpsoralen (TMP). The mutagenized DNA sequence was the EcoR1-Hind III restriction fragment in the vicinity of the Tcsup(R) promoter. Two classes of mutants were obtained. One exhibited a high level of Tc resistance (40-60 ..mu..g/ml) but still lower than the wild-type. Interestingly, these showed no sequence alterations at all in the vicinity of the TMP-reacted fragment. The other class of mutants exhibited low levels of drug resistance (< 20 ..mu..g/ml) and two of those that were sequenced were found to contain a 15-base pair insertion to the right of the original Hind III site. Under the conditions used, psoralen ...
Full text: The excessive use of chemical pesticides to control agricultural pests is becoming alarming. The objective of this study is to search for biopesticides of plant origin that could be used to control one of the major pest of fruit production; the Mediterranean fruit fly (Ceratitis capitata Wied.). A colony of the Lebanese wild strain of this insect was reared under laboratory condition to provide biological material. The insecticidal activity of the essential oils extracted from aromatic plants in Lebanon was assessed. The tested plants are: Foeniculum vulgare, Thymbra spicata, Artemisia herba alba, Origanum syriacum, Ruta chalepensis, Lavandula stoechas, Salvia fruticosa, Mentha microphylla, Juniperus oxycedrus, Rosmarinus officinalis, Myrtus communis, Laurus nobilis and Ocimum gratissimum. Results show that essential oils isolated from F. vulgare, T. spicata, A. herba alba, O. syriacum and R. chalepensis have promising insecticidal potential. (author)
A risk-neutral method is always used to price and hedge contingent claims in complete market, but another method based on utility maximization or risk minimization is wildly used in more general case. One can find all kinds of special risk measure in literature. In this paper, instead of using market modified risk measure, we use a kind of risk measure induced by g_\\Gamma-solution or the minimal solution of a Constrained Backward Stochastic Differential Equation (CBSDE) directly when constraints on wealth and portfolio process comes to our consideration. Such g_\\Gamma-solution and the risk measure generated by it is well defined on appropriate space under suitable conditions. We adopt the inf-convolution of convex risk measures to solve some optimization problem. A dynamic version risk measures defined through g_\\Gamma-solution and some similar results about optimal problem can be got in our new framework and by our new approach.
The production of cellulolytic enzymes from fungi has been extensively studied. Several mutants of Trichoderma reesei were selected. Most of the studies were carried out on T. reesei, T. viride, T. harzianum, Penicillium funiculosum, Altemaria alternata. Aspergillus phoenicis, A. ustus, A. tamarii, A. japonicus, and A. niger. T. koningii is one of the most active producers of the so-called C, factor, which is indispensable for the rapid and extensive attack on crystal-line cellulose. However, Trichodenna is known to excrete only small amounts of {beta}-glucosidase. Therefore, Trichoderma is supplemented with {beta}-glucosidase from Aspergillus to increase the saccharification rate of cellulose to glucose as the main sugar. Induction of mutations in Trichodenna spp. rather than T. viride as a tool for the enhancement of {beta}-glucosidase activity was reported. Unfortunately, T. reesei is a poor producer of {beta}-glucosidase. On the other hand, T. harzianum M{sub 5}, a mutant that was ...
Protocols were standardized for in vitro propagation of several elite and diverse banana accessions using shoot tip explants. Tissue culture raised plants were field planted at multiple locations. Studies were undertaken for the induction of mutations using multiple shoot cultures of six selected cultivars, Shreemanti (AAA), Basrai (AAA), Lal Kela (AAA), Rasthali (AAB), Karibale Monthan (ABB) and a wild diploid (BB). These shoot cultures were irradiated at different doses of gamma rays (0-100 Gy) and subcultured thrice (up to M_1V_3) to separate shimeras, followed by induction of rooting (M_1V_4). In general, the rate of multiplication had a negative association with the dose of gamma rays. Enhanced multiplication of shoots was noticed at lower doses. The proliferation of shoots was arrested beyond 50 Gy and a dose of 70 Gy was completely lethal for all the genotypes studied. The rooted plantlets were hardened in the green house and in the early stages of field ...
Homozygous transgenic cotton (Gossypium hirsutum L.) plants that accumulated glycinebetaine (GB) in larger quantities were more tolerant to salt than wild-type (WT) plants. Four transgenic lines, namely 1, 3, 4, and 5, accumulated significantly higher levels of GB than WT plants did both before and after salt stress. At 175 and 275?mM NaCl, seeds of all the transgenic lines germinated earlier and recorded a higher final germination percentage, and the seedlings grew better, than those of the WT. Under salt stress, all the lines showed some characteristic features of salt tolerance, such as higher leaf relative water content (RWC), higher photosynthesis, better osmotic adjustment (OA), lower percentage of ion leakage, and lower peroxidation of the lipid membrane. Levels of endogenous GB in ...
This study describes the role of "inflammatory" oxidized (Ox) phospholipids in regulation of rahU (PA0122) expression and biofilm formation in Pseudomonas aeruginosa (383) wild type (rahU+) and rahU mutant (rahU-) strains. Functional analysis of RahU protein from P. aeruginosa in presence of Ox-phospholipids show: (a) LysoPC modulates RahU gene/and protein expression in rahU+ cells; (b) rahU promoter activity is increased by lysoPC and inhibited by PAPC, Ox-PAPC and arachidonic acid; the latter inhibitory effect can be reversed by lysoPC, which was enzymatically derived from PAPC; (c) biofilm formation increased in rahU- cells as compared to rahU+; and (d) inhibition of rahU promoter activity by PAPC and AA (but not lysoPC) showed significantly augmented biofilm formation in rahU+ but not ...
The Fifteenmile Basin Habitat Improvement Project is an ongoing multi-agency effort to improve habitat in the Fifteenmile drainage and increase production of the depressed wild, winter steelhead run. Cooperating agencies include the Oregon Department of Fish and Wildlife, USDA Forest Service. USDA Soil Conservation Service and Bonneville Power Administration. in consultation with the Confederated Tribes of Warm Springs. The Oregon Department of Fish and Wildlife is administering project work on state and private lands and the U.S.D.A. Forest Service is administering project work on National Forest land. Project work on the Forest has been sub-divided into four components; (1) Ramsey Creek, (2) Eightmile Creek, (3) Fifteenmile Creek, and (4) Fivemile Creek. Forest Service activities in the Fifteenmile basin during 1988 involved habitat improvement work on Ramsey Creek, continuation of physical and biological monitoring, collection of spawning survey information, and ...
The goal of the Fifteenmile Creek Habitat Improvement project is to improve wild winter steelhead habitat in the Fifteenmile Creek Basin. This goal was addressed under the Columbia River Basin Fish and Wildlife Program, Measure 703 (c) (1) - Action Item 4.2. Construction of fish habitat structures was completed on approximately 3.5 miles of Eightmile Creek and on approximately 8.5 miles of Fifteenmile Creek. A total of 15,435 yds of rock was placed to improve rearing and spawning habitat, and to improve water quality and reduce siltation. Riparian protection fencing was completed on 1.8 miles of Dry Creek, approximately 1.75 miles on Eightmile Creek, and 4.75 miles on Fifteenmile Creek. Approximately 10 miles of stream will receive protection from the 8 miles of new fencing installed and 6.5 miles of abandoned fence repaired. Irrigation withdrawal screens were installed at six irrigation pump withdrawals. These screens functioned satisfactorily during the ...
A good knowledge of medfly population dynamics is necessary to control it. Population dynamics are being studied by means of the Jackson trap with trimedlure attractant (male, sex-pheromone). Weekly sampling of wild medfly populations was started in June at Aabde and in August at Sour and Jbeil. Trap surveys of medfly density will be carried for at least one year before sterile males releases are started against a long-established infestation. Since populations can vary with season and in different parts of an infested area. Knowledge of this variation is needede to determine when releases should start, because they should begin, just after or during a population decline. Field evaluation will include, ecological data on medfly population distribution number, host preference, and medfly overwintering. Surveys of medfly adults and larvae, host species, and phenology, temperature and medfly history, all will be used to guide the control program, specifically the ...
A comprehensive dataset of NDV genome sequences was evaluated using bioinformatics to characterize the evolutionary forces affecting NDV genomes. Despite evidence of recombination in most genes, only one event in the fusion gene of genotype V viruses produced evolutionarily viable progenies. The codon-associated rate of change for the six NDV proteins revealed that the highest rate of change occurred at the fusion protein. All proteins were under strong purifying (negative) selection; the fusion protein displayed the highest number of amino acids under positive selection. Regardless of the phylogenetic grouping or the level of virulence, the cleavage site motif was highly conserved implying that mutations at this site that result in changes of virulence may not be favored. The coding sequence of the fusion gene and the genomes of viruses from wild birds displayed higher yearly rates of change in virulent viruses than in viruses of low virulence, suggesting that an ...
The Drosophila melanogaster tissue-specific transcription factor NTF-1 was originally identified in vitro as a protein that could bind to and activate transcription from the Dopa decarboxylase (Ddc) gene. A structure-function analysis of NTF-1 led to the identification of a discrete amino-terminal activation domain. Here, we report that an NTF-1 mutant lacking the activation domain acts as a trans-dominant inhibitor of NTF-1 activation in tissue culture cells by forming inactive heterodimers with the full-length protein. Ectopically expressing this dominant-negative protein or the full-length protein in developing Drosophila embryos leads to dire developmental consequences. Overexpressing the trans-dominant NTF-1 leads to lethality, while overexpressing full-length NTF-1 results in both lethality and morphogenetic defects. Our results suggest that both the activity and the regulation of NTF-1 are critical for viability and proper development of the fly.ImagesFig. 1Fig. 2Fig. 3
Based on studies on the genetic and molecular basis of Shigella flexneri invasive properties, we have constructed and evaluated a double mutant of S. flexneri serotype 5 for utilization as a live attenuated oral vaccine against shigellosis. The first mutation, icsA, blocks intracellular spread of bacteria as well as cell-to-cell infection. It affects the capacity of the invasive pathogen to form large abscesses in epithelia. The second mutation, iuc, eliminates production of the siderophore aerobactin thus impairing growth of the bacterium within tissues. This double mutant, SC5700 appeared safe when administered intragastrically to macaque monkeys as three doses (5 x 10(10) c.f.u. each) at weekly intervals. Protection against a challenge by the wild type isolate (M90T) was observed 4 weeks after the last vaccine inoculation. Duration of carriage was considerably reduced as compared to the control group in which all animals had developed severe dysentery. ...
The photobiology of mouse melanocyte lines with different pigment genotypes was studied by measuring colony-forming ability after irradiation. The cell lines were wild-type black (melan-a) and the mutants brown (melan-b) and albino (melan-c). Four lamps emitting various UV wavelengths were used. These were germicidal (UVC, 200-280 nm), 82.3% output at 254 nm, TL01 (UVB, 280-320 nm), 64.2% at 310-311 nm, FS20, broadband with peak output at 312 nm and Alisun-S (UVA, 320-400 nm), broadband with peak output at 350-354 nm. Appropriate filtration reduced the contaminating UVC to nonlethal levels for the longer waverange lamps. It is clear from these studies that, in pigment cells, monochromatic results cannot predict polychromatic responses and that cell death from solar irradiations is a complex phenomenon that depends on more than DNA damage. (author).
The malM locus of Streptococcus pneumoniae was cloned into one of the two PstI sites of the multicopy S. pneumoniae plasmid pMV158. To eliminate chromosomal transformants in the simultaneous selection for tetracycline resistance (coded by pMV158) and maltose utilization, the host cells contained a chromosomal deletion of the mal gene cluster. Two clones were isolated; one with a 3.3 kb insert (pLS70) which behaved like wild type with respect to maltose utilization, and another with a 2.9 kb insert (pLS69) which behaved as though it contained a down promoter mutation. Preliminary mapping of these clones by restriction analysis placed the 0.4kb deletion on a HindIII fragment in the interior of the chromosomal insert. The recombinant plasmids were able to transform over 50% of a recipient population to Mal/sup +/. Enzyme measurements of the clones indicated an overproduction of amylomaltase, constituting up to 10% of the total cellular protein, and supported the ...
The potential of the dried yeast, wild-type Schizosaccharomyces pombe, to remove Ni(II) ion was investigated in batch mode under varying experimental conditions including pH, temperature, initial metal ion concentration and biosorbent dose. Optimum pH for biosorption was determined as 5.0. The highest equilibrium uptake of Ni(II) on S. pombe, q e, was obtained at 25??C as 33.8?mg?g?1. It decreased with increasing temperature within a range of 25?50??C denoting an exothermic behaviour. Increasing initial Ni(II) concentration up to 400?mg?L?1 also elevated equilibrium uptake. No more adsorption took place beyond 400?mg?L?1. Equilibrium data fitted better to Langmuir model rather than Freundlich model. Sips, Redlich?Peterson, and Kahn isotherm equations modelled the investigated system with a...
We describe the isolation and genetic characterization of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced mutations in the phage P22 mnt repressor gene cloned in plasmid pBR322. Mutations in the mnt repressor gene or its operator on this plasmid, pPY98, confer a tetracycline resistance phenotype, whereas the wild-type plasmid confers tetracycline sensitivity. Cells carrying pPY98 were briefly exposed to MNNG to give 20 to 40% survival and a 50- to 100-fold increase in tetracycline-resistant cells. DNA sequence analysis showed that 29 to 30 MNNG-induced mutations were GC-to-AT transitions and one was an AT-to-GC transition. About 80% of the mutations are in three hotspots. This mutation spectrum is consistent with the proposed mechanism of mutagenic action of MNNG, which involves mispairing of an alkylated base, O/sup 6/-methylguanine. The mnt gene may be a useful target for determining mutagenic specificity at the nucleotide level because (i) forward ...
Structural instability of wild-type fibroblast growth factor (FGF)-1 and its dependence on exogenous heparin for optimal activity diminishes its potential utility as a therapeutic agent. Here we evaluated FGFC, an FGF1:FGF2 chimeric protein, for its receptor affinity, absolute heparin-dependence, stability and potential clinical applicability. Using BaF3 transfectants overexpressing each FGF receptor (FGFR) subtype, we found that, like FGF1, FGFC activates all of the FGFR subtypes (i.e., FGFR1c, FGFR1b, FGFR2c, FGFR2b, FGFR3c, FGFR3b and FGFR4) in the presence of heparin. Moreover, FGFC activates FGFRs even in the absence of heparin. FGFC stimulated keratinocytes proliferation much more strongly than FGF2, as would be expected from its ability to activate FGFR2b. FGFC showed greater struct...
In order to study the neurobiological basis of seasonal changes in hippocampal structure and function, the rate of cell proliferation was examined in male and female wild meadow voles captured during different seasons. We found that the number of [3H]thymidine-labeled cells varied across the seasons and across sex in the meadow vole. Non-breeding female meadow voles had a higher rate of cell proliferation and cell death than males captured during either season or breeding females. These seasonal changes in the female meadow vole were associated with both fluctuating levels of adrenal steroids and gonadal steroids. Estradiol level was highly correlated with both the number of [3H]thymidine-labeled cells and the number of pyknotic cells in female meadow voles, with high levels of estradiol being associated with low levels of cell proliferation and cell death. Corticosterone level was associated with the number of [3H]thymidine-labeled cells in the hilus of female ...
S100A4, a member of the Ca(2+)-activated S100 protein family, regulates the motility and invasiveness of cancer cells. Moreover, high S100A4 expression levels correlate with poor patient survival in several cancers. Although biochemical, biophysical, and structural data indicate that S100A4 is a noncovalent dimer, it is unknown if two functional S100A4 monomers are required for the productive recognition of protein targets and the promotion of cell invasion. To address this question, we created covalently linked S100A4 dimers using a glycine rich flexible linker. The single-chain S100A4 (sc-S100A4) proteins exhibited wild-type affinities for calcium and nonmuscle myosin-IIA, retained the ability to regulate nonmuscle myosin-IIA assembly, and promoted tumor cell invasion when expressed in S100A4-deficient colon carcinoma cells. Mutation of the two calcium-binding EF-hands in one monomer, while leaving the other monomer intact, caused a 30-60-fold reduction in ...
Vascular endothelial growth factor (VEGF) is a mitogen that is critically involved in vasculogenesis, angiogenesis, and hematopoiesis. However, what and how transcription factors participate in the regulation of vegf gene expression are not fully understood. Here we report the cloning and sequencing of the zebrafish vegf promoter which revealed that the promoter contains a number of bone morphogenetic protein (BMP)-activated Smad binding elements (SBE), implicating Smad1 and Smad5 in the regulation of BMP-induced expression of vegf. Electrophoretic mobility shift assays of adding recombinant Smad proteins to the SBE-containing DNA oligonucleotides that represent portions of zebrafish vegf promoter resulted in mobility shift of the oligonucleotides. These changes demonstrate potential interactions between Smad1/5 and the vegf promoter. Reporter activity assays using the wild-type or SBE-deleted vegf promoters to drive the luciferase reporter gene expression revealed ...
With the advent of subgenomic hepatitis C virus (HCV) replicons, studies of the intracellular steps of the viral replication cycle became possible. These RNAs are capable of self-amplification in cultured human hepatoma cells, but save for the genotype 2a isolate JFH-1, efficient replication of these HCV RNAs requires replication enhancing mutations (REMs), previously also called cell culture adaptive mutations. These mutations cluster primarily in the central region of non-structural protein 5A (NS5A), but may also reside in the NS3 helicase domain or at a distinct position in NS4B. Most efficient replication has been achieved by combining REMs residing in NS3 with distinct REMs located in NS4B or NS5A. However, in spite of efficient replication of HCV genomes containing such mutations, they do not support production of infectious virus particles. By using the genotype 1b isolate Con1, in this study we show that REMs interfere with HCV assembly. Strongest impairment of virus formation ...
Estimated kokanee (Oncorhynchus nerka) abundance in Lake Pend Oreille was 4.3 million during September 1986. This estimate was similar to 1985 and indicates continued suppression of the kokanee population since initial decline in the late 1960s. Atypically high survival of wild fry resulted in similar fry recruitment in 1986 as 1985, whereas hatchery-reared fry contributed only 8% to total fry recruitment as a result of low post-release survival (3%). Fry released into the Clark Fork River from Cabinet Gorge Hatchery had very low survival during emigration to Lake Pend Oreille, resulting from poor flow conditions and potentially high predation. Fry survival during emigration was twice as high during nighttime flows of 16,000 cfs than 7,800 cfs. Emigration also was faster during higher flows. Several marks were tested to differentially mark fry release groups to help determine impacts of flow and other factors on fry survival. Survival of fry marked with ...
The biobaler was originally developed for short-rotation willow plantations, but can currently harvest a wide range of woody crops with a basal diameter up to 150 mm. The biobaler is an alternate approach to harvest woody crops as round bales, generally 1.2 m wide by 1.5 m diameter. In addition to harvesting trees, it can improve management of wild brush, forest understory vegetation and encroaching small trees on abandoned land. It allows easy handling, storage and transportation to sites where the biomass can be used for energy use or other applications. This paper reported on a study that was conducted in the fall of 2009 in which a third generation biobaler was used on 8 different sites across Minnesota, notably Waseca, Madelia, Faribault, Afton, Ogilvie, Hinckley, Aurora and Hibbing. A total of 160 bales were harvested from these sites. The average bale mass was 466 kg and average bale density was 296 kg/m{sup 3}. The moisture content averaged 44.9 per cent ...
In mice, coat pigmentation requires a stem cell (SC) system in which the survival, proliferation, and differentiation of melanocytes (MCs) are regulated by microenvironments in hair follicles (HFs). In vitro systems are required to analyze the behavior of single melanocyte stem cells (MCSCs) and their potential to form SC systems in vivo. We describe here an experimental system for the isolation, self-renewal, and differentiation of MCSCs, as well as an in vivo reconstitution assay for assessing their potential. Using Dct(tm1(Cre)Bee)/CAG-CAT-GFP mice, we show that, in the presence of stem cell factor and basic fibroblast growth factor and the XB2 feeder cell line, purified MCSCs can undergo clonogenic proliferation, resulting in c-Kit(low) side scatter(low) cells. In culture, these cells maintain their capacity to differentiate and reconstitute an MCSC system in HFs. As these cells are present in the upper part of the HF near the bulge region, express only low levels of housekeeping ...
The Stardust sample return capsule will return to Earth in January 2006 with primitive debris collected from Comet 81P/Wild-2 during the fly-by encounter in 2004. In addition to the cometary particles embedded in low-density silica aerogel, there will be microcraters preserved in the Al foils (1100 series; 100 {micro}m thick) that are wrapped around the sample tray assembly. Soda lime spheres ({approx}49 {micro}m in diameter) have been accelerated with a Light Gas Gun into flight-grade Al foils at 6.35 km s{sup -1} to simulate the capture of cometary debris. The experimental craters have been analyzed using scanning electron microscopy (SEM) and x-ray energy dispersive spectroscopy (EDX) to locate and characterize remnants of the projectile material remaining within the craters. In addition, ion beam induced secondary electron imaging has proven particularly useful in identifying areas within the craters that contain residue material. Finally, high-precision ...
No living analogue exists for the hypothetical early hominid hard/tough-seed, coarse-root-eating, and bone-crushing masticatory adaptation. To investigate possible microdamage/microwear to dental enamel caused by such usage, puncture-crushing experiments were carried out on single human teeth, using an Instron compression apparatus on the following six test materials: Makapansgat Limeworks chert (e.g., taphonomy), fresh steer longbone, mongongo nuts, Grewia berries, Carob beans, and wild-onion bulbs. Pairs of extracted unworn third molars were utilized, with one tooth acting as the control. The teeth were mounted, ultrasonically cleaned, and two-stage replicas made with a vinyl polysiloxane elastomer and araldite epoxy resin. After Instron loading and materials failure (1.2-395.0 kg) the test items and the crowns were prepared for comparison with scanning electron microscopy and dispersive x-ray elemental analysis and mapping. The results revealed that although ...
Mutations in transcription factors hepatocyte nuclear factors (HNF)-1#alpha# and HNF-1#beta# cause maturity-onset diabetes of the young (MODY) types 3 and 5, respectively. HNF-1#alpha# and HNF-1#beta# mutations are well studied in some tissues, but the mechanism by which HNF-1#alpha# and HNF-1#beta# mutations affect sucrase-isomaltase (SI) transcription in the small intestine is unclear. We studied the effects of 13 HNF-1#alpha# mutants and 2 HNF-1#beta# mutants on human SI gene transcription, which were identified in subjects with MODY3 and MODY5, respectively. Transactivation activity of 11 HNF-1#alpha# and 2 HNF-1#beta# mutants was significantly lower than that of wild (wt)-HNF-1#alpha# and wt-HNF-1#beta#. Furthermore, in co-expression studies with mutant (mu)-HNF-1#alpha#/ wt-HNF-1#beta# and wt-HNF-1#alpha#/mu-HNF-1#beta#, the combination of mu-HNF-1#alpha# (P379fsdelCT and T539fsdelC)/wt-HNF-1#beta# impaired SI transcription, but the others were not remarkably ...
The detection of rare mutations has many important applications, including risk assessment of drugs and chemicals, measuring environmental exposures to genotoxins, and cancer cell detection. A sensitive genotypic selection method has been developed that combines two different mutant allele selection techniques, MutEx enrichment and allele-specific competitive blocker PCR (ACB-PCR). This method was developed and evaluated for the detection of a CAA --> AAA mutation at codon 61 of the mouse H-ras gene. The MutEx enrichment is based on MutS binding to a mismatched basepair in heteroduplex DNA. The bound MutS protects the mutant allele from degradation during subsequent exonuclease treatment. ACB-PCR preferentially amplifies a mutant allele in a PCR reaction using a primer that has more mismatches to the wild-type allele than the mutant allele. By combining these two approaches, the codon 61 mutation was detected at mutant fractions as low as 1 in 10(7). This ...
Neuronal transcription factors play vital roles in the specification and development of neurons, including dopaminergic (DA) neurons. Mutations in the gene encoding the purine biosynthetic enzyme hypoxanthine-guanine phosphoribosyltransferase (HPRT) cause the resulting intractable and largely untreatable neurological impairment of Lesch-Nyhan disease (LND). The disorder is associated with a defect in basal ganglia DA pathways. The mechanisms connecting the purine metabolic defect and the central nervous system (CNS) phenotype are poorly understood but have been presumed to reflect a developmental defect of DA neurons. We have examined the effect of HPRT deficiency on the differentiation of neurons in the well-established human (NT2) embryonic carcinoma neurogenesis model. We have used a retrovirus expressing a small hairpin RNA (shRNA) to knock down HPRT gene expression and have examined the expression of a number of transcription factors essential for neuronal differentiation and ...
Adenine DNA glycosylase catalyzes the glycolytic removal of adenine from the promutagenic A {center_dot} oxoG base pair in DNA. The general features of DNA recognition by an adenine DNA glycosylase, Bacillus stearothermophilus MutY, have previously been revealed via the X-ray structure of a catalytically inactive mutant protein bound to an A:oxoG-containing DNA duplex. Although the structure revealed the substrate adenine to be, as expected, extruded from the DNA helix and inserted into an extrahelical active site pocket on the enzyme, the substrate adenine engaged in no direct contacts with active site residues. This feature was paradoxical, because other glycosylases have been observed to engage their substrates primarily through direct contacts. The lack of direct contacts in the case of MutY suggested that either MutY uses a distinctive logic for substrate recognition or that the X-ray structure had captured a noncatalytically competent state in lesion recognition. To gain further ...
We present an abundance analysis based on high-resolution spectra of 10 stars selected to span the full range in metallicity in the Ursa Minor (UMi) dwarf spheroidal (dSph) galaxy. We find that [Fe/H] for the sample stars ranges from -1.35 to -3.10 dex. Combining our sample with previously published work for a total of 16 luminous UMi giants, we establish the trends of abundance ratios [X/Fe] as functions of [Fe/H] for 15 elements. In key cases, particularly for the #alpha#-elements, these trends resemble those for stars in the outer part of the Galactic halo, especially at the lowest metallicities probed. The neutron-capture elements show an r-process distribution over the full range of Fe metallicity reached in this dSph galaxy. This suggests that the duration of star formation in the UMi dSph was shorter than in other dSph galaxies. The derived ages for a larger sample of UMi stars with more uncertain metallicities also suggest a population dominated by uniformly old (#approx#13 ...
Heterochromatin constitutes a significant portion of the genome in higher eukaryotes; approximately 30% in Drosophila and human. Heterochromatin contains a high repeat DNA content and a low density of protein-encoding genes. In contrast, euchromatin is composed mostly of unique sequences and contains the majority of single-copy genes. Genetic and cytological studies demonstrated that heterochromatin exhibits regulatory roles in chromosome organization, centromere function and telomere protection. As an epigenetically regulated structure, heterochromatin formation is not defined by any DNA sequence consensus. Heterochromatin is characterized by its association with nucleosomes containing methylated-lysine 9 of histone H3 (H3K9me), heterochromatin protein 1 (HP1) that binds H3K9me, and Su(var)3-9, which methylates H3K9 and binds HP1. Heterochromatin formation and functions are influenced by HP1, Su(var)3-9, and the RNA interference (RNAi) pathway. My thesis project investigates how ...
Our long-term goal is to enable highly productive plant species to extract, resist, detoxify, and/or sequester toxic heavy metal pollutants as an environmentally friendly alternative to physical remediation methods. We have focused this phytoremediation research on soil and water-borne ionic and methylmercury. Mercury pollution is a serious world-wide problem affecting the health of human and wild-life populations. Methylmercury, produced by native bacteria at mercury-contaminated wetland sites, is a particularly serious problem due to its extreme toxicity and efficient biomagnification in the food chain. We engineered several plant species (e.g., Arabidopsis, tobacco, canola, yellow poplar, rice) to express the bacterial genes, merB and/or merA, under the control of plant regulatory sequences. These transgenic plants acquired remarkable properties for mercury remediation. (1) Transgenic plants expressing merB (organomercury lyase) extract methylmercury from their ...
1. Contest rules The European Space Agency (ESA) is launching a public competition to find the most suitable names for its four Cluster II space weather satellites. The quartet, which are currently known as flight models 5, 6, 7 and 8, are scheduled for launch from Baikonur Space Centre in Kazakhstan in June and July 2000. Professor Roger Bonnet, ESA Director of Science Programme, announced the competition for the first time to the European Delegations on the occasion of the Science Programme Committee (SPC) meeting held in Paris on 21-22 February 2000. The competition is open to people of all the ESA member states (*). Each entry should include a set of FOUR names (places, people, or things from history, mythology, or fiction, but NOT living persons). Contestants should also describe in a few sentences why their chosen names would be appropriate for the four Cluster II satellites. The winners will be those which are considered most suitable and relevant for the Cluster II mission. ...