Column chromatographic analysis of barbiturates in their dosage forms. II. secobarbital, amobarbital, and pentobarbital.

A general method for the analysis of barbiturates, using column parition chromatography, was extended to the assay of secobarbital, amobarbital, and pentobarbital. A solution of the barbiturate constitutes the immobile phase in the chromatographic system. It is eluted with ether-isooctane (1+9) and passed onto a K3PO4 column, which retains the barbiturate while extraneous materials are washed out. The barbiturate is removed from the column with etherisooctane (3+1), extracted from the eluate with NH3, and measured spectrophotometrically.     

Volumetric method for determing traces of organic chlorine in lipids.

The official AOAC method for determining chlorine in cake flour lipids has been modified to determine chlorine in lipids of animal carcasses cooled with chlorinated water. The sensitivity of the method has been improved 1000-fold to attain a detection limit of 2 ppm Cl in lipids. Recovery of the fine silver chloride precipitate was improved through centrifugation rather than filtration. The official method also did not take into consideration that when the silver chloride precipitate is exposed to light, it is converted to free chlorine and silver. Recovery of organic chlorine ranged from 84.6% at 4.1 ppm to 100.1% at 100 ppm. A number of samples of commercially chlorinated beef, pork, and chicken and laboratory-chlorinated chicken were analyzed by this method. In all cases where the level of chlorination was sufficient to result in a chlorine level in lipids in excess of 2 ppm, the chlorine content of the carcass lipids was measurable.     

The genome of black cottonwood, Populus trichocarpa (Torr. & Gray)

We report the draft genome of the black cottonwood tree, Populus trichocarpa. Integration of shotgun sequence assembly with genetic mapping enabled chromosome-scale reconstruction of the genome. More than 45,000 putative protein-coding genes were identified. Analysis of the assembled genome revealed a whole-genome duplication event; about 8000 pairs of duplicated genes from that event survived in the Populus genome. A second, older duplication event is indistinguishably coincident with the divergence of the Populus and Arabidopsis lineages. Nucleotide substitution, tandem gene duplication, and gross chromosomal rearrangement appear to proceed substantially more slowly in Populus than in Arabidopsis. Populus has more protein-coding genes than Arabidopsis, ranging on average from 1.4 to 1.6 putative Populus homologs for each Arabidopsis gene. However, the relative frequency of protein domains in the two genomes is similar. Overrepresented exceptions in Populus include genes associated with lignocellulosic wall biosynthesis, meristem development, disease resistance, and metabolite transport.     

Zinc mediation of the binding of human growth hormone to the human prolactin receptor

Human growth hormone (hGH) elicits a diverse set of biological activities including lactation that derives from binding to the prolactin (PRL) receptor. The binding affinity of hGH for the extracellular binding domain of the hPRL receptor (hPRLbp) was increased about 8000-fold by addition of 50 micromolar ZnCl2. Zinc was not required for binding of hGH to the hGH binding protein (hGHbp) or for binding of hPRL to the hPRLbp. Other divalent metal ions (Ca2+, Mg2+, Cu2+, Mn2+, and Co2+) at physiological concentrations did not support such strong binding. Scatchard analysis indicated a stoichiometry of one Zn2+ per hGH.hPRLbp complex. Mutational analysis showed that a cluster of three residues (His18, His21, and Glu174) in hGH and His188 from the hPRLbp (conserved in all PRL receptors but not GH receptors) are probable Zn2+ ligands. This polypeptide hormone.receptor "zinc sandwich" provides a molecular mechanism to explain why nonprimate GHs are not lactogenic and offers a molecular link between zinc deficiency and its association with altered functions of hGH.     

Microsatellite genetic variation between and within farmed and wild Atlantic salmon (Salmo salar) populations

Genetic diversity between three farmed and four wild populations of Atlantic salmon from Ireland and Norway were analysed using 15 microsatellite markers. High levels of polymorphism were observed over all populations with the average number of alleles and average heterozygosity at 17.8 and 0.70, respectively. Farmed salmon showed less genetic variability than wild salmon in terms of allelic diversity but not necessarily in terms of overall heterozygosity. Between farmed populations significant differences were observed in expected heterozygosity suggesting that more intensive breeding practices may have resulted in a further erosion of genetic variability. Phylogenetic analysis using either populations or individuals as nodes show a clustering of populations into two groups, farmed and wild. This suggests that founder effects and subsequent selection have had more effect on the genetic differentiation between these strains than geographical separation. This technology has great potential for use in aquaculture situation where levels of genetic variation could be monitored and inbreeding controlled in a commercial breeding progra.     

Endosomal proteolysis of the Ebola virus glycoprotein is necessary for infection

Ebola virus (EboV) causes rapidly fatal hemorrhagic fever in humans and there is currently no effective treatment. We found that the infection of African green monkey kidney (Vero) cells by vesicular stomatitis viruses bearing the EboV glycoprotein (GP) requires the activity of endosomal cysteine proteases. Using selective protease inhibitors and protease-deficient cell lines, we identified an essential role for cathepsin B (CatB) and an accessory role for cathepsin L (CatL) in EboV GP-dependent entry. Biochemical studies demonstrate that CatB and CatL mediate entry by carrying out proteolysis of the EboV GP subunit GP1 and support a multistep mechanism that explains the relative contributions of these enzymes to infection. CatB and CatB/CatL inhibitors diminish the multiplication of infectious EboV-Zaire in cultured cells and may merit investigation as anti-EboV drugs.     

Outcome of equids with clinical signs of West Nile virus infection and factors associated with death

OBJECTIVE: To determine outcome of equids in the western United States with clinical signs of West Nile virus (WNV) infection and identify factors associated with risk of death in infected equids. DESIGN: Cross-sectional study. ANIMALS: 484 equids in Nebraska and Colorado. PROCEDURE: Owners of 484 equids with laboratory-confirmed West Nile virus infection in Nebraska and Colorado were contacted by telephone, and a questionnaire was used to obtain information on signalment, management, clinical signs, date of disease onset, duration of disease, WNV vaccination status, and health status at the time of the interview. RESULTS: 137 of 482 (28.4%) animals died or were euthanatized. Ataxia, lethargy, muscle fasciculations, and weakness were the most common clinical signs of disease. Animals > or = 3 years old were more likely to die than were animals < or = 2 years old. Unvaccinated equids were twice as likely to die as were animals that had been vaccinated at least once prior to the onset of disease. Animals that were recumbent and unable to rise were 78 times as likely to die as were animals that never lost the ability to rise. Females were 2.9 times as likely to die as males. Two hundred seventy-one of 339 (79.9%) animals that survived recovered fully; mean duration of disease for these animals was 22.3 days. CONCLUSIONS AND CLINICAL RELEVANCE: Among equids with WNV infection, age, vaccination status, an inability to rise, and sex were associated with the risk of death.     

Population biology and epidemiology of plant virus epidemics: from tripartite to tritrophic interactions

Natural enemies have long been used in biological control programs to mitigate the damage caused by herbivory. Many herbivorous insect species also act as plant virus vectors, enabling virus transmission from plant to plant and hence disease development in a plant population. Whilst an intuitive assumption would be to expect a decrease in vector numbers to lead to subsequent reductions in virus transmission, recent evidence suggests that introduction of natural enemies (parasitoids and predators) may in some cases increase plant virus transmission while at the same time reducing vector numbers. In this paper we review the evidence for plant-virus-vector-natural enemy interactions, the signalling mechanisms involved and their implications for virus transmission, and show how a modelling approach can assist in identifying the key parameters and relationships involved in determining the disease outcome. A mathematical model linking the population dynamics of a vector-parasitoid system with virus transmission was used to investigate the effects of virus inoculation and acquisition rates, parasitoid attack rate and vector aggregation on disease dynamics across a wide range of parameter value combinations. Virus spread was found to increase with enhanced inoculation, acquisition and parasitoid attack rate but decrease with high levels of vector aggregation.