Population structure,dispersal and colonization history of the garden bumblebee <Emphasis Type="Italic">Bombus hortorum</Emphasis> in the Western Isles of Scotland

New methods of analysing genetic data provide powerful tools for quantifying dispersal patterns and reconstructing population histories. Here we examine the population structure of the bumblebee Bombus hortorum in a model island system, the Western Isles of Scotland, using microsatellite markers. Following declines in other species, B. hortorum is the only remaining long-tongued bumblebee species found in much of Europe, and thus it is of particular ecological importance. Our data suggest that populations of B. hortorum in western Scotland exist as distinct genetic clusters occupying groups of nearby islands. Population structuring was higher than for other bumblebee species which have previously been studied in this same island group (F_st = 0.16). Populations showed significant isolation by distance. This relationship was greatly improved by using circuit theory to allow dispersal rates to differ over different landscape features; as we would predict, sea appears to provide far higher resistance to dispersal than land. Incorporating bathymetry data improved the fit of the model further; populations separated by shallow seas are more genetically similar than those separated by deeper seas. We argue that this probably reflects events following the last ice age when the islands were first colonized by this bee species (8,500–5,000 ybp), when the sea levels were lower and islands separated by shallow channels would have been joined. In the absence of significant gene flow these genetic clusters appear to have since diverged over the following 5,000 years and arguably may now represent locally adapted races, some occurring on single islands.     

Development of a diagnostic test system for detecting soluble staphylococcal antigens by an immunoenzyme method

ELISA is used for detecting the soluble staphylococcal antigen in patients with purulent septic infections. The optimum conditions for the assay have been established: the dose of staphylococcal gamma globulin for plate sensitization should be 5.0-10.0 micrograms/ml, the pH of the buffer solution 9.6-10.0, the time and temperature of incubation 18-20 hours at 4 degrees C or 5 hours at 37 degrees C. The possibility of using plates manufactured in the USSR has been shown. The sensitivity of the above diagnostic test system is 0.005 microgram/ml.     

Long noncoding RNA PTENP1 affects the recovery of spinal cord injury by regulating the expression of miR-19b and miR-21

Exosomes derived from differentiated P12 cells and MSCs were proved to suppress apoptosis of neuron cells, and phosphatase and tensin homolog pseudogene 1 (PTENP1) was reported to inhibit cell proliferation. In this study, we aimed to investigate the role of PTENP1 in the process of post-spinal cord injury (SCI) recovery, so as to evaluate the therapeutic effects of exosomes derived from MSCs transfected with PTENP1 short hairpin RNA (shRNA), as a type of novel biomarkers in the treatment of SCI. Electron microscopy was used to observe the morphology of different exosomes. Real-time polymerase chain reaction and western blot, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays, flow cytometry, Nissl staining, immunohistochemistry assay, and terminal deoxynucleotidyl transferase dUTP nick end labeling assay were conducted to investigate and validate the underlying molecular signaling pathway. PTENP1-shRNA downregulated PTENP1 and PTEN while upregulating miR-21 and miR-19b. PTENP1-shRNA also accelerated cell apoptosis and reduced cell viability. In addition, PTENP1 reduced the miR-21 and miR-19b expression by directly targeting miR-21 and miR-19b. Meanwhile, both miR-21 and miR-19b reduced the expression of PTEN by directly targeting the 3′-untranslated region of PTEN. Furthermore, PTEN level and apoptosis index of neuron cells was the highest in the SCI group, while the treatment with exosomes+PTENP1-shRNA reduced the PTEN expression to a level similar to that in the sham group. Finally, PTENP1 inhibited miR-21 and miR-19b expression but upregulated PTEN expression. The upregulation of miR-21/miR-19b also suppressed the apoptosis of neuron cells by downregulating the PTEN expression. PTENP1 is involved in the recovery of SCI by regulating the expression of miR-19b and miR-21, and exosomes from PTENP1-shRNA-transfected cells may be used as a novel biomarker in SCI treatment.     

Simple reversed-phase high-performance liquid chromatography quantitation of ganciclovir in human serum and urine

A fast, simple, and cost-effective HPLC method for the quantitation of the antiviral drug ganciclovir is described. The serum samples are extracted with perchloric acid and neutralized with potassium phosphate buffer, and urine samples are diluted with distilled water. A reversed-phase column with isocratic elution by 15 mM potassium phosphate buffer (pH 2.5) containing 0.25% acetonitrile is used to separate ganciclovir; quantitation is by UV absorbance at 254 nm. Total turnaround time is 22 min; more than 3000 samples can be run on a single column without loss of peak quality. The limit of quantitation is 0.05 μg/ml. Recoveries varied from 91 to 10% with coefficients of variation ranging from 0.387 to 7.95%.     

The cortical response in Xenopus laevis ova

A dependence on extracellular calcium has been demonstrated for fertilization and the cortical response to pricking in Xenopus ova. Neither event occurred in calcium-free solutions or in the presence of divalent cation chelating agents. The calcium-sensitive phase of the cortical response to pricking in dejellied eggs was restricted to the 5–10 sec immediately following the activation stimulus; the initial phase of activation was not calcium dependent. In contrast, the cortical response in dejellied Xenopus ova exposed to the chemical activating agents, urethan or methyl urethan, was independent of extracellular calcium. Experimental evidence was presented for the involvement of a direct, nonpropagated cortical reaction in response to urethan stimulation as opposed to a propagated reaction in response to pricking. A cortical response in dejellied eggs was not induced spontaneously by high concentrations of potassium, and the prick response was unaffected by inhibitors of energy transfer processes. Molecular mechanisms operative in the initiation and propagation of the cortical response in animal eggs have been discussed.