Analysis and visualization of H7 influenza using genomic,evolutionary and geographic information in a modular web service

We have reported previously on use of a web‐based application, Supramap ( http://supramap.org ) for the study of biogeographic, genotypic, and phenotypic evolution. Using Supramap we have developed maps of the spread of drug‐resistant influenza and host shifts in H1N1 and H5N1 influenza and coronaviruses such as SARS. Here we report on another zoonotic pathogen, H7 influenza, and provide an update on the implementation of Supramap as a web service. We find that the emergence of pathogenic strains of H7 is labile with many transitions from high to low pathogenicity, and from low to high pathogenicity. We use Supramap to put these events in a temporal and geospatial context. We identify several lineages of H7 influenza with biomarkers of high pathogenicity in regions that have not been reported in the scientific literature. The original implementation of Supramap was built with tightly coupled client and server software. Now we have decoupled the components to provide a modular web service for POY ( http://poyws.org ) that can be consumed by a data provider to create a novel application. To demonstrate the web service, we have produced an application, Geogenes ( http://geogenes.org ). Unlike in Supramap, in which the user is required to create and upload data files, in Geogenes the user works from a graphical interface to query an underlying dataset. Geogenes demonstrates how the web service can provide underlying processing for any sequence and metadata database. © The Willi Hennig Society 2012.     

The dark-colour-inducing neurohormone of locusts in relation to an albino mutant of Schistocerca gregaria

In the albino mutant of an Okinawa strain of Locusta migratoria (L.) (Orthoptera: Acrididae), albinism is caused by the absence of the dark‐colour‐inducing neurohormone (DCIN), which is present in the corpora cardiaca (CC) of normally coloured phenotypes. This study tests whether the absence of DCIN is responsible for albinism in an albino mutant of another locust, Schistocerca gregaria (Forsk.) (Orthoptera: Acrididae). This seemed feasible because a single Mendelian unit controls albinism in both species. However, implantation of CC, or injection of an extract of CC, from albino donors of S. gregaria, induce dark coloration in crowded nymph recipients of the Okinawa albino mutant of L. migratoria, as effectively as do implanted CC, or injections of extract of CC, from normal phenotype donors of S. gregaria. Therefore, DCIN is present in the albino mutant of S. gregaria, and consequently, the albinism in this mutant is not caused by its absence. Implantation of CC, or injection of extracts of CC, from albino donors of S. gregaria to conspecific albino nymphs does not induce darkening. Only extremely high doses of synthetic DCIN injected into albino nymphs of S. gregaria are effective, inducing some darkening. The dose to induce such darkening in albino nymphs of S. gregaria is 50 nmol, ≈ 5 × 10⁶ times higher than that (10 femtomol) needed to induce equivalent darkening in nymphs of the Okinawa albinos of L. migratoria. The results are discussed and some possible explanations of the observed effects outlined.     

A critical evaluation of the use of filipin-permeabilized rat hepatocytes to study functions of the endoplasmic reticulum in situ

We have used transmission (TEM) and scanning electron microscopy (SEM) and leakage of lactate dehydrogenase (LDH; EC 1.1.1.27) to evaluate two published procedures which use filipin to render isolated rat hepatocytes permeable to ionic substrates. Cells treated by the procedure of Jorgenson and Nordlie retained less than 10 per cent of their LDH. TEM revealed severe damage to the internal structure of these cells, which included swelling, disintegration and extensive vesicularization of the endoplasmic reticulum (ER). Hepatocytes treated with filipin by the procedure of Gankema et al. retained 65-75 per cent of their LDH and displayed incomplete but highly variable permeability to Trypan blue. SEM revealed the loss of microvilli, other signs of swelling, and the presence of large lesions in the plasma membrane. TEM revealed signs of cell swelling, but the nuclei and the mitochondria were only moderately altered. The rough ER was not swollen, but significant fragmentation was evident and characteristic stacks of lamellar ER were never seen. We conclude that useful information about the functions of the ER in situ cannot be obtained from studies of filipin-treated cells. Our results indicate that retention of LDH is not a sufficient criterion of preservation of cell morphology and that staining with Trypan blue may significantly underestimate the permeability of cells to small ionic metabolites.     

Antiepileptic effects of amphetamine may require GABA (benzodiazepine) activity

Secondary components of visual evoked potentials (slow negative wave-SNW, and photically-evoked sensory afterdischarge-SAD) are known to be precursors of experimentally activated wave-spike discharges, similar to wave-spikes of petit mal epilepsy. Both SNW and SAD may be potently suppressed wither by amphetamine or GABAergic compounds such as diazepam and sodium valproate. A hypothesis was tested in the present study, that amphetamine-induced suppression of wave-spike discharges may require GABA-benzodiazepine activity for its expression.Electrocortical activity was recorded and averaged in unrestrained albino rats with chronically implanted epicortical electrodes. SNW and SAD obtained in habituated rats in the predrug state were potently suppressed by amphetamine (1 mg/kg, i.p.). Fifteen minutes after amphetamine injection, a challenging drug (metrazol, picrotoxin, convulsant benzodiazepine, Ro 5-3663, or imidazodiazepine, Ro 15-1788) was administered intraperitoneally. Subconvulsive doses of metrazol (10 mg/kg) reversed amphetamine suppression; imidazodiazepine (20 mg/kg) and picrotoxin (1.5 mg/kg) reliably opposed the SNW suppression; convulsant benzodiazepine, Ro 5-3663 (2 mg/kg), showed modest and nonsignificant effect in the same direction. It is proposed that the antiepileptic potency of amphetamine may be associated with its ability, apparently via modulatory effect of norepinephrine, to facilitate the activation of benzodiazepine-GABA receptors.     

Limestone gravel as growth medium in hydroponics

Summary Plants were grown in gravel beds of basalt or limestone. With the standard nutrient solution yields for the plants grown in the limestone gravel were very much reduced and the plants were chlorotic. This chlorosis and reduced yields remained even if the gravel was acid or water washed beforehand, or if it was pretreated in high phosphate concentrations.High yields, comparable to those obtained in basalt beds were obtained if phosphate and iron were added in small amounts (0.1mM P, 1 mg/1 Fe) at each irrigation. This technique to avoid lime induced chlorosis can be easily and economically accomplished in hydroponics. It proved successful here with lettuce, cucumber, tomato and eggplants.     

Antibody Response of Animals to Mycoplasma pneumoniae Measured by Latex Agglutination

The standardization, application, and usefulness of latex agglutination for Mycoplasma pneumoniae antibody titration were investigated and compared with tetrazolium reduction inhibition and complement fixation. The sera of guinea pigs and monkeys reacted in a specific fashion, whereas rabbit serum required pretreatment to eliminate its nonspecific agglutinin. Human serum given such pretreatment still contained a nonspecific agglutinin. The latex agglutination procedure compared favorably with complement fixation and metabolic inhibition tests for evaluating vaccine antigenicity.     

Titration and Neutralization of Poliovirus in Micro Tissue Culture Under Increased Carbon Dioxide

Modification of the micro tissue culture technique, including incubation under increased CO(2), resulted in prolongation of the viability of the cells. As a consequence, satisfactory titrations of poliovirus and of poliovirus antiserum have been achieved by the micro method. The technique offers a number of advantages over conventional methods.     

Pseudohypoparathyroidism type Ia: two new heterozygous frameshift mutations in exons 5 and 10 of the Gsα gene

Pseudohypoparathyroidism type Ia (PHP-Ia) is a hereditary disease characterized by resistance to PTH and other hormones that act via cAMP. Patients have deficient activity of Gs, the subunit of the G protein, which couples hormone receptors to stimulation of adenylate cyclase. We describe two new mutations discovered in two sporadic patients with PHP-Ia. Using genomic DNA, we have amplified exons 2–13 of the Gs gene (GNAS1) by PCR, and sequenced the resulting products. Both patients had Albright's hereditary osteodystrophy, resistance to multiple hormones, and deficient Gs activity. In the first patient, a deletion of a C in exon 5 at codon 115 was found. In the second patient, an insertion of a C in exon 10 at codon 267 was detected. Both these heterozygous mutations cause frameshift, and predict decreased production of Gs. This report adds two new Gs mutations to the known ten mutations recently described.     

Remote sensing of chlorophyll in Lake Kinneret using highspectral-resolution radiometer and Landsat TM: spectral features of reflectance and algorithm development

High-resolution reflectance spectra in the range of 400–850nm were obtained from Lake Kinneret during a period when densepopulations of the dinoflagellate Peridinium gatunense dominatedthe phytoplankton. Chlorophyll (Chl) concentrations ranged from5.1 to 185 mg m^–3 and from 2.4 to 187.5 mg m^–3 inthe samples of two independent experiments. The most prominentfeatures of the reflectance spectra were: (i) a wide minimumfrom 400 to 500 nm; (ii) a maximum at 550–570 nm, whichdid not surpass 3% in samples with high Chl concentration (>20mgm^–3), indicating a strong absorption by pigments in thegreen range of the spectrum; (iii) a minimum at 676 nm; thiswas {small tilde}1% and was almost insensitive to variationin Chl concentration >10 mg m^–3; (iv) a maximum reflectanceshowed near 700 nm; its magnitude and position were highly dependenton chlorophyll concentration. High-spectral-resolution datawere used as a guideline for selection of the most suitablespectral bands for chlorophyll remote sensing. Models were devised,based on the calculation of the integrated area above the baselinefrom 670 to 850 nm and the reflectance maximal height withinthis range. Some algorithms already used m previous studieswere tested and showed a plausible degree of accuracy when appliedto the current data base. However, novel models devised in thisstudy improved substantially the accuracy of Chl estimationby remotely sensed data, by reducing the estimation error from>11 to 6.5 mg m^–3 Those models were validated by anindependent data set where Chl concentration ranged over twoorders of magnitude. The use of three relatively narrow spectralbands was sufficient for Chl mapping in Lake Kinneret. Therefore,a relatively simple sensor, measuring only a few bands willbe employed in future applications for Chl monitoring in inlandwaters. Radiometric data were also used to simulate radiancesin the channels of TM Landsat and to find the algorithm forChl assessment. The ratio of channel 4 to channel 3 was usedand enabled Chl estimation with an error of <15mg m^–3This algorithm was employed to map Chl in the entire area ofLake Kinneret with 10 gradations.