The prevalence of Fasciola hepatica in its snail intermediate host determined by DNA probe assay

Accurate snail intermediate host infection prevalence data have the potential to be extremely useful in determining seasonal transmission dynamics of Fasciola hepatica. Because the microscopic techniques currently used lack the sensitivity and specificity necessary to obtain meaningful infection prevalence data, we developed a highly accurate and efficient DNA probe assay. The assay has a sensitivity of 100%, a specificity of >99%, easily detects a single miracidia and does not cross-hybridize with DNA of Fascioloides magna, Paramphistomum liorchis or Heterobilharzia americana, trematodes that share the same intermediate host and enzootic range as Fasciola hepatica. Using this assay, we determined the prevalence of F. hepatica in its snail intermediate host, Fossaria cubensis, during the second year of a 2-year study on the epizootiology of Fasciola hepatica in Florida. The overall infection prevalence of snails assayed in this study (n = 5246) was 1.5% and ranged from 0.1% to 3.1% for individual cattle ranches. Additionally, infection prevalence differed significantly for successive size groupings of snails, varying from 0% for 1-mm snails to 18.5% for 9- and 10-mm snails. The accuracy and efficiency of the DNA probe assay reported here for determining snail infection prevalence offers an inexpensive alternative to tracer animal studies for determining the epizootiology of F. hepatica.     

Ethanol Specifically Inhibits NMDA Receptors with Affinity for Ifenprodil in the Low Micromolar Range in Cultured Cerebellar Granule Cells

Abstract: The effect of ethanol on the intracellular Ca²⁺ concentration response to NMDA in rat cerebellar granule cells grown in low or high KCI concentrations has been studied using image analysis. The cells grown in low KCI displayed high sensitivity for glycine. The subtype-selective antagonist ifenprodil inhibited the response with high (in the low micromolar range) and low (in the high micromolar range) potency. Ethanol affected the high-potency component in these cultures. In cells grown in high KCI the glycine sensitivity was lower, and a low potency for ifenprodil (high micromolar) dominated. These cells were not significantly sensitive to ethanol. The results indicate that the component displaying potency for ifenprodil in the low micromolar range with properties of the NR2B subunit is the target for ethanol action on the NMDA receptor.     

A colorimetric method for the determination of submicrogram quantities of protein

A simple procedure for the measurement of submicrogram quantities of protein is described which can be used without interference from most common reagents. Protein-containing solutions are spotted on glass fiber filters, washed with trichloroacetic acid, and stained with Coomassie blue. The filters are destained, and the protein-bound colorant is eluted and measured in a spectrophotometer at 590 nm. The response is linear to 5 μg of protein per filter, and as little as 0.1 μg per filter can be accurately determined.     

The S49 Kin- cell line transcribes and translates a functional mRNA coding for the catalytic subunit of cAMP-dependent protein kinase

The S49 mouse lymphoma mutant cell line Kin- is resistant to the cytotoxic effects of elevated cAMP levels, has no detectable cAMP-dependent protein kinase activity, and has depressed levels of cAMP-binding regulatory subunits. We demonstrate that although the Kin- cell line lacks detectable catalytic subunit protein, these cells express wild-type levels of mRNA for both C alpha and C beta catalytic subunit isoforms. Translation of C alpha mRNA appears to be normal in the Kin- cell, based on the observation that C alpha mRNA associates with large polyribosomes in both wild-type and Kin- cells. We cloned the C alpha cDNA from Kin- cells and show that its transient expression in another cell type leads to activation of a cAMP-sensitive luciferase reporter gene, suggesting that functional C alpha protein is made. In addition to having catalytic activity, the C alpha subunit from Kin- cells is inhibited in the presence of mouse RI alpha regulatory subunit, indicating that formation of the holoenzyme complex is normal. We suggest that the mutation responsible for the Kin- phenotype is in a cellular component that directly or indirectly causes Kin- catalytic subunit protein to be degraded rapidly.     

Continuous-variable quantification of dermatoglyphic whorl patterns: a statistical study of angular measurements

Core(s) and triradii of dermatoglyphic whorl patterns were joined together to form triangles. Base angles of these triangles were measured (in degrees). The tangent angle at the lower edge of a ridge crossing the core-triradius line was also measured in degrees on each side of the whorl. Significant differences and similarities of these angles were investigated for unrelated Caucasian males and females by the use of Student's t- and Pearson's r-tests. Angular findings were related to the corresponding information provided by ridge counts. Similarities and differences between males and females are described.     

gA and gB glycoproteins of herpes simplex virus type 1: two forms of a single polypeptide.

Utilizing a combination of preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis and sodium dodecyl sulfate-hydroxylapatite column chromatography, we have separated and purified the gA and gB glycoproteins of the major virus-specific glycoprotein region from herpes simplex virus type 1-infected cells. By using purified antigen preparations, antisera specific to each of these glycoproteins were produced. Immunoprecipitation from detergent extracts of infected cells and radioimmune precipitation of the purified antigens have shown that the anti-gA and anti-gB sera each recognize both the gA and the gB glycoproteins. The anti-gA serum was also shown to neutralize virus despite the presence of only minute quantities of the gA glycoprotein in virions. Pulse-chase studies have indicated that the gA and gB glycoproteins are synthesized from a common precursor polypeptide. Together, these data demonstrate that the gA and gB glycoproteins of herpes simplex virus type 1 are antigenically similar but not identical and probably represent two different forms of the same polypeptide which differ in their degree of glycosylation.     

Accumulation of polychlorinated biphenyls in turbot(Scophthalmus maximus) from seawater sediments and food

Juvenile turbot,Scophthalmus maximus (L.), were exposed to 0.58 μg 1⁻¹ Aroclor 1254 in seawater, to sediments containing 100, 60 and 1 ppm or fed with cockle containing 20 ppm PCB (polychlorinated biphenyls). Concentration factors for liver and muscle were 10⁴ and 10³, respectively, for uptake of PCB from seawater. Contamination of muscle was similar to that of sediments containing 1 and 60 ppm PCB to which turbot were exposed, but less than the 20 ppm in their experimental diet. Contamination of flatfish in the North Sea area is compared with the levels of PCB in the flounder,Platichthys flesus (L.), in the River Thames and predictable values for uptake of PCB from different pathways discussed.     

Synthesis of enkephalins by guinea-pig striatum in vitro.

A method for obtaining incorporation of labelled amino acids into enkephalins of guinea-pig striatum in vitro and isolating the labelled enkephalins is described. The incorporation of [3H]tyrosine increases with time after a delay of 2 h. Cycloheximide added during this time, but not at later stages, inhibits incorporation. The results indicate that the enkephalins are produced locally, probably by ribosomal synthesis. The lag period may be due to the time required for the synthesis of precursors and their conversion to the enkephalins.     

DNA-negative temperature-sensitive mutants of herpes simplex virus type 1: patterns of viral DNA synthesis after temperature shift-up.

Temperature-sensitive mutants of herpes simplex virus type 1 belonging to four DNA- complementation groups exhibited two distinct patterns of viral DNA synthesis after shift-up to the nonpermissive temperature. In cultures infected with mutants belonging to complementation groups A, C, and D, little or no viral DNA was synthesized after shift-up. In cultures infected with a mutant in complementation group B, nearly normal amounts of viral DNA were synthesized after shift-up.