Regulation of transient receptor potential melastatin 8 (TRPM8) channel activity by its short isoforms.

One important mechanism of membrane ion channels regulation involves their non-functional isoforms generated by alternative splicing. However, knowledge of such isoforms for the members of transient receptor potential (TRP) superfamily of ion channels remains quite limited. This study focuses on TRPM member, TRPM8, which functions as a cold receptor in sensory neurons, but is also expressed in tissues not exposed to ambient temperatures, as well as in cancer tissues. We report the cloning from prostate cancer cells of new short-splice variants of TRPM8, termed short TRPM8a (sM8a) and short TRPM8ß (sM8ß). Our results show that both variants are in a closed configuration with the C-terminal tail of the full-size TRPM8 chan-nel, resulting in stabilization of its closed state and thus reducing both its cold sensitivity and its activity. Our findings, therefore, uncover a new mode of the regulation of TRPM8 channel by its splice variants.

Testate amoebae as indicators of hydroseral change: an 8,500 year record from Mer Bleue Bog, eastern Ontario, Canada.

Testate amoebae have been used widely as a proxy of hydrological change in ombrotrophic peatlands, although their response to abiotic controls in other types of mire and fenland palaeo-environments is less well understood. This paper examines the response of testate amoebae to hydroseral and other environmental changes at Mer Bleue Bog, Ontario, Canada, a large ombrotrophic peatland, which evolved from a brackish-water embayment in the early Holocene. Sediments, plant macrofossils and diatoms examined from a 5.99 m core collected from the dome of the bog record six stages of development: i) a quiet, brackish-water riverine phase (prior to ca. 8500 cal BP); ii) a shallow lake (ca. 8500–8200 cal BP); iii) fen (8200–7600 cal BP); iv) transitional mire (7600–6900 cal BP); v) pioneer raised mire (6900–4450 cal BP); and vi) ombrotrophic bog (4450 cal BP-present).

Testate amoebae, notably small (<25 µm diameter) specimens of Centropyxis aculeata type, first appear in low abundances in sediments ascribed to the lacustrine phase. Diatoms from the same horizons record a shallowing in water depth, a decline in salinity and the development of emergent macrophytic vegetation, which may have provided favourable conditions for testate amoeba colonization. The testate amoeba communities of the inferred fen phase are more diverse and include centropyxids, cyclopyxids, Arcellidae and Hyalospheniidae, although the assemblages show some differences to those recently reported in modern European fen environments. The Fen–Bog Transition (FBT) is also dominated by C. aculeata type. The change in testate amoeba communities around this key transition is apparent in the results of Detrended Correspondence Analysis (DCA), and appears to reflect a latent nutrient gradient and a secondary moisture gradient. DCA analyses of plant macrofossil remains around the FBT show a similar trend, although the sensitivity of the two proxies to the inferred environmental changes differs. Comparisons with other regional mid-Holocene peatland records confirm the important influence of reduced effective precipitation on the testate amoeba communities during the initiation and development of Sphagnum-dominated, raised bog communities.

False-positive PCR results linked to administration of seasonal influenza vaccine

False-positive PCR results usually occur as a consequence of specimen-to-specimen or amplicon-to-specimen contamination within the laboratory. Evidence of contamination at time of specimen collection linked to influenza vaccine administration in the same location as influenza sampling is described. Clinical, circumstantial and laboratory evidence was gathered for each of five cases of influenza-like illness (ILI) with unusual patterns of PCR reactivity for seasonal H1N1, H3N2, H1N1 (2009) and influenza B viruses. Two 2010 trivalent influenza vaccines and environmental swabs of a hospital influenza vaccination room were also tested for influenza RNA. Sequencing of influenza A matrix (M) gene amplicons from the five cases and vaccines was undertaken. Four 2009 general practitioner (GP) specimens were seasonal H1N1, H3N2 and influenza B PCR positive. One 2010 GP specimen was H1N1 (2009), H3N2 and influenza B positive. PCR of 2010 trivalent vaccines showed high loads of detectable influenza A and B RNA. Sequencing of the five specimens and vaccines showed greatest homology with the M gene sequence of Influenza A/Puerto Rico/8/1934 H1N1 virus (used in generation of influenza vaccine strains). Environmental swabs had detectable influenza A and B RNA. RNA detection studies demonstrated vaccine RNA still detectable for at least 66 days. Administration of influenza vaccines and clinical sampling in the same room resulted in the contamination with vaccine strains of surveillance swabs collected from patients with ILI. Vaccine contamination should therefore be considered, particularly where multiple influenza virus RNA PCR positive signals (e.g. H1N1, H3N2 and influenza B) are detected in the same specimen.

Vegetation responses to rapid climatic changes during the last deglaciation 13,500-8,000 years ago on southwest Andoya, arctic Norway

The late-glacial vegetation development in northern Norway in response to climate changes during the Allerod, Younger Dryas (YD), and the transition to the Holocene is poorly known. Here we present a high-resolution record of floral and vegetation changes at lake Lusvatnet, south-west Andoya, between 13500 and 8000 cal b.p. Plant macrofossil and pollen analyses were done on the same sediment core and the proxy records follow each other very closely. The core has also been analyzed using an ITRAX XRF scanner in order to check the sediment sequence for disturbances or hiatuses. The core has a good radiocarbon-based chronology. The Saksunarvatn tephra fits very well chronostratigraphically. During both the Allerod and the Younger Dryas time-periods arctic vegetation prevailed, dominated by Salix polaris associated with many typically arctic herbs such as Saxifraga cespitosa, Saxifraga rivularis and Oxyria digyna. Both periods were cold and dry. Between 12450 and 12250 cal b.p. during the Younger Dryas chronozone, the assemblage changed, particularly in the increased abundance of Papaver sect. Scapiflora and other high-Arctic herbs, suggesting the development of polar desert vegetation mainly as a response to increased aridity. After 11520 cal b.p. a gradually warmer and more oceanic climate initiated a succession to dwarf-shrub vegetation and the establishment of Betula woodland after 1,000 years at c. 10520 cal b.p. The overall late-glacial aridity contrasts with oceanic conditions in southern Norway and is probably related to sea-ice extent.

The incidences of trisomy 8, trisomy 9 and D20S108 deletion in polycythaemia vera: an analysis of blood granulocytes using interphase fluorescence in situ hybridization:an analysis of blood granulocytes using interphase fluorescence in situ hybridization

We have used interphase fluorescence in situ hybridization (IFISH) to detect trisomy 8, trisomy 9 and 20q deletion in circulating granulocytes from patients with polycythaemia vera (PV). Out of 64 PV patients, 15 (23%) exhibited an abnormality. Two patients had trisomy 9, three had trisomy 8 and 10 patients had hemizygous deletion of D20S108 (a locus in the 20q common deleted region). Aberrant nuclei ranged from 10% to 80% in these 15 cases. There was no correlation between the presence of a marker and sex, age, interval between presentation and IFISH analysis, neutrophil or platelet count or therapy. Conventional marrow cytogenetic karyotype results were available in 23 cases and there was concurrence between these and blood IFISH in 16 cases (13 normal and three with 20q/D20S108 deletion by both methods). Three patients with D20S108 deletion by IFISH were normal by previous marrow cytogenetic testing and four cases with 20q deletion by previous marrow cytogenetics had normal blood granulocytes according to IFISH. Thus, we confirm that trisomies 8 and 9 and deletion of 20q are diagnostically useful markers of PV. IFISH analysis of blood granulocytes is a practical method for detecting these markers, but as an adjunct to, not as a substitute for, conventional marrow cytogenetics.